Details! (Draft 2.0, subject to revision)

Details! (Draft 2.0, subject to revision)
Quality Assurance Project Plan
Wolf Run Watershed Based Plan
EPA 319(h) Grant No. C9994861-09
Prepared By:
Third Rock Consultants, LLC
2526 Regency Road Suite 180, Lexington KY 40503
859-977-2000
Prepared For:
Lexington-Fayette Urban County Government
Department of Environmental Protection
200 East Main Street, Lexington, KY 40507
859-425-2800
and
Kentucky Department for Environmental Protection
Division of Water
200 Fair Oaks Lane
Frankfort, KY 40601
502-564-3410
Date: April 11, 2011
Revision Date: Revision No.: 0
This Page Intentionally Blank
SECTION A – PROJECT MANAGEMENT
A1. Title and Approval Sheet
Quality Assurance Project Plan
for Wolf Run Watershed Based Plan
April 12, 2011
Steve Evans / QAPP Author and Project Quality
Assurance Officer, Third Rock Consultants, LLC
Date
Susan Bush / Project Manager, Lexington-Fayette
Urban County Government
Date
David Price / Laboratory Manager, Lexington-Fayette
Urban County Government
Date
Ken Cooke / Volunteer Coordinator, Friends of Wolf
Run
Date
Brook Shireman / Project Manager, Kentucky
Division of Water
Date
Lisa A. Hicks / Quality Assurance Officer, Kentucky
Division of Water
Date
________________________________________________________________________
Wolf Run Watershed
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________________________________________________________________________
Wolf Run Watershed
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Revision History
This page documents the revisions over time to this document. The most recent iteration
should be listed in the first space, with consecutive versions following. Signatures may be
required for revised documents.
Date of Revision
04/12/11
Page(s)/Section(s)
Revised
All
Revision Explanation
Original preparation date of document
________________________________________________________________________
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A2. Table Of Contents
SECTION A – PROJECT MANAGEMENT................................................................. 3
A1. Title and Approval Sheet........................................................................................... 3
A2. Table Of Contents ..................................................................................................... 6
A3. Distribution List ......................................................................................................... 9
A4. Project / Task Organization .................................................................................... 10
A5. Project Definition / Background............................................................................. 11
A6. Project/Task Description......................................................................................... 13
A6.1. General Overview of Project............................................................................... 13
A6.1.1. Karst Hydrograph Characterization ............................................................ 17
A6.1.2. Conductivity Survey ................................................................................... 17
A6.1.3. Benthic Macroinvertebrate Collection ........................................................ 17
A6.1.4. Watershed Habitat Assessments ................................................................. 18
A6.1.5. Hydrogeomorphic Assessment ................................................................... 18
A6.1.6. Water Quality Monitoring........................................................................... 19
A6.2. Project Timetable ................................................................................................ 19
A7. Data Quality Objectives (DQOs) and Criteria for Measurement Data .............. 22
A7.1. Data Quality Objectives ...................................................................................... 22
A7.2. Data Quality Indicators ....................................................................................... 24
A7.2.1. Precision....................................................................................................... 24
A7.2.2. Accuracy ...................................................................................................... 25
A7.2.3. Representativeness ....................................................................................... 26
A7.2.4. Completeness ............................................................................................... 26
A7.2.5. Comparability............................................................................................... 27
A7.2.6. Sensitivity..................................................................................................... 27
A8. Training Requirements............................................................................................ 27
A9. Documentation and Records ................................................................................... 27
A9.1. Field Documentation and Records...................................................................... 27
A9.2. Laboratory Documentation and Records ............................................................ 28
A9.3. Quality Documentation and Final Reports.......................................................... 29
SECTION B. - DATA GENERATION AND ACQUISITION................................... 30
B1. Sampling Process Design ......................................................................................... 30
B1.1. Sampling Site and Reach Locations.................................................................... 30
B1.2. Sampling Design Rationale................................................................................. 32
B1.2.1. Karst Hydrograph Characterization ............................................................ 32
B1.2.2. Conductivity Survey.................................................................................... 34
B1.2.3. Benthic Macroinvertebrate Collection ........................................................ 34
B1.2.4. Watershed Habitat Assessments.................................................................. 34
B1.2.5. Hydrogeomorphic Assessment.................................................................... 34
B1.2.6. Water Quality Monitoring........................................................................... 35
B2. Sampling Methods.................................................................................................... 36
B2.1. Sampling Equipment ........................................................................................... 36
B2.2. Sampling Methods............................................................................................... 36
B2.2.1. Karst Hydrograph Characterization ............................................................ 36
B2.2.2. Conductivity Survey.................................................................................... 39
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B2.2.3. Benthic Macroinvertebrate Collection ........................................................ 40
B2.2.4. Watershed Habitat Assessments.................................................................. 44
B2.2.5. Hydrogeomorphic Assessment.................................................................... 48
B2.2.6. Water Quality Monitoring........................................................................... 50
B3. Sample Handling and Custody Requirements ...................................................... 53
B3.1. Chain-of-Custody ................................................................................................ 53
B3.2. Sample Handling and Transport.......................................................................... 54
B3.3. Sample Labeling.................................................................................................. 54
B3.4. Sample Designation............................................................................................. 55
B4. Analytical Methods Requirements ......................................................................... 55
B5. Quality Control Requirements................................................................................ 56
B5.1. Field Quality Controls......................................................................................... 56
B5.2. Macroinvertebrate Taxonomic Quality Controls ................................................ 56
B5.3. Chemical Laboratory Quality Controls ............................................................... 57
B6. Instrument / Equipment Testing, Inspection, Calibration, and Maintenance
Requirements................................................................................................................... 57
B7. Data Management ................................................................................................... 58
SECTION C – ASSESSMENT AND OVERSIGHT ................................................... 59
SECTION D – DATA VALIDATION AND USABILITY.......................................... 61
D1. Data Review, Validation and Verification ............................................................. 61
D2. Validation and Verification Methods..................................................................... 62
D3. Reconciliation with User Requirements and Data Quality Objectives ............... 63
SECTION E. - REFERENCES AND CITATIONS..................................................... 63
List of Exhibits
Exhibit 1 – Project Sampling Sites ................................................................................... 14
Exhibit 2 – Karst System Hydraulic Monitoring .............................................................. 15
Exhibit 3 – Watershed Habitat Assessment Streams ........................................................ 33
List of Tables
Table 1 – Project Roles and Responsibilities.................................................................... 11
Table 2 – Monitoring Activity Overview ......................................................................... 13
Table 3 – Sampling Equipment Summary ........................................................................ 16
Table 4 – Project Schedule ............................................................................................... 21
Table 5 – Summary of Data Quality Objectives and Standard Operating Procedures ..... 23
Table 6 – Acceptance Criteria for Water Chemistry and In-Situ Measurements ............. 24
Table 7 – Completeness Goals of Sampling Activities .................................................... 26
Table 8 – Sampling Locations .......................................................................................... 31
Table 9 – Habitat Assessment Stream Segments.............................................................. 32
Table 10 – Sample Preservation and Hold Time .............................................................. 36
Table 11 – Summary of Sampling Methods for Macroinvertebrates ............................... 41
Table 12 – Field Equipment Calibration and Maintenance .............................................. 57
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Table 13 – Watershed Assessment and Management Reports ......................................... 60
Table 14 – Data Qualifiers and Response......................................................................... 62
List of Figures
Figure 1 – Wolf Run Watershed Monitoring Organization Chart .................................... 10
Figure 2 – Measurement of Discharge Through Sub-Sectional measurements................ 37
List Of Appendices
Appendix A – Datasheets
Appendix B – Standard Operating Procedures
________________________________________________________________________
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A3. Distribution List
The following individuals will receive the approved Quality Assurance Project Plan
(QAPP) and any subsequent revisions.
Name
Address
Steve Evans
Third Rock Consultants, LLC
Gerry Fister
Third Rock Consultants, LLC
Marcia Wooton
Third Rock Consultants, LLC
Bert Remley
Third Rock Consultants, LLC
Jennifer Shelby
Third Rock Consultants, LLC
Susan Bush
Lexington-Fayette Urban
County Government
David Price
Lexington-Fayette Urban
County Government
2526 Regency Road, Suite 180
Lexington, KY 40503
2526 Regency Road, Suite 180
Lexington, KY 40503
2526 Regency Road, Suite 180
Lexington, KY 40503
2526 Regency Road, Suite 180
Lexington, KY 40503
2526 Regency Road, Suite 180
Lexington, KY 40503
Jason Backus
Kentucky Geological Survey
Ken Cooke
Friends of Wolf Run
Brooke Shireman
KY Division of Water
Lisa A. Hicks,
KY Division of Water
Phone
Number
E-mail Address
859-977-2000
[email protected]
859-977-2000
[email protected]
859-977-2000
[email protected]
859-977-2000
[email protected]
859-977-2000
[email protected]
200 East Main Street,
Lexington, KY 40507
859-425-2800
[email protected]lfucg.com
301 Lisle Industrial Avenue
Lexington, Kentucky 40511
859-425-2415
[email protected]
859-323-0555
[email protected]
859-940-8234
[email protected]
502-564-3410
[email protected]
502-564-3410
[email protected]
228 Mining & Mineral
Resources Building
University of Kentucky
Lexington, KY 40506-0107
639 Cardinal Lane
Lexington, Kentucky 40503
200 Fair Oaks Lane
Frankfort, KY 40601
200 Fair Oaks Lane
Frankfort, KY 40601
________________________________________________________________________
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A4. Project / Task Organization
This Quality Assurance Project Plan (QAPP), prepared by Third Rock Consultants, LLC
(Third Rock), is to be reviewed and approved by the Kentucky Division of Water
(KDOW) and the Lexington-Fayette Urban County Government (LFUCG). This QAPP
establishes the planning, implementation, and assessment procedures necessary to meet
the minimum data quality objectives (DQOs) for the monitoring of the Wolf Run
watershed.
Third Rock is committed to producing quality data that will assist the KDOW and
LFUCG in obtaining the information necessary to facilitate the development of a Wolf
Run Watershed Based Plan. This QAPP is designed to provide a complete plan for
achieving all project data quality objectives. However, effective communication is
essential to ensure that all parties properly implement the plan. Any project related
quality feedback, questions, or concerns should be communicated to the project
administrator or quality manager to facilitate appropriate analysis and resolution.
The implementation of the monitoring plan requires the effective operation of the project
team. Figure A-1, Wolf Run Watershed Monitoring Organizational Chart, identifies the
parties that comprise the Project Team and the lines of authority and communication
under which this team operates. The specific roles and responsibilities of each key party
are documented in Table 1.
FIGURE 1 – WOLF RUN WATERSHED MONITORING ORGANIZATION
CHART
KDOW
QA Officer
Lisa Hicks
KDOW
Project Manager
Brooke Shireman
LFUCG
Grantee Project Manager
Susan Bush
Third Rock
Project Manager
Gerry Fister
Friends of Wolf Run
Sampling Coordinator
Ken Cooke
Third Rock
Sampling Coordinator
Marcia Wooton
Third Rock
Macroinvertebrate
Laboratory
Bert Remley
Third Rock
QA Manager
Steve Evans
LFUCG
Laboratory Director
David Price
Kentucky Geological Survey
Laboratory Services
Manager
Jason Backus
________________________________________________________________________
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TABLE 1 – PROJECT ROLES AND RESPONSIBILITIES
Name
Brooke Shireman
KDOW
Lisa Hicks
KDOW
Susan Bush
LFUCG
Gerry Fister
Third Rock
Project Title
KDOW Project
Manager
KDOW QA Officer
Grantee Project
Manager
Project Manager
Steve Evans
Third Rock
QA Manager
Marcia Wooton
Third Rock
Sampling
Coordinator
Ken Cooke
Friends of Wolf Run
David Price
LFUCG
Jason Backus
Kentucky Geological
Survey
Volunteer Sampling
Coordinator
Bert Remley
Third Rock
Laboratory Director
Responsibility
Administratively oversees data collection planning
and reviews data
Administratively oversees QAPP conditions; may
assist in review of data
Oversight of project
Project scheduling and coordination to meet time
line and budget goals
Development of the QAPP; ensure QAPP
compliance, conduct audits, review and approve
all data generated; preparation of QA reports
Coordination of water quality sample collection
events with volunteer staff; bottle preparation and
labeling, laboratory communication on hold times
and data results
Coordination of volunteer sampling activities;
training volunteers per QAPP specifications
Chemistry laboratory analysis for water quality
samples
Laboratory Services
Manager
Chemistry laboratory analysis for water quality
samples
Macroinvertebrate
Laboratory Chief
Taxonomist
Oversee and conduct field biological sampling;
biological data review; identification of benthic
samples; oversee macroinvertebrate quality
assurance
A5. Project Definition / Background
Wolf Run was first listed as impaired for swimming use (non-support) in the 1998 303(d)
list of Kentucky impaired waters. This impaired status has remained since that time with
additional impairments (partial support of warmwater aquatic habitat use and non-support
of secondary contact use) being identified in subsequent years (KDOW 2010a). The
impairment of Wolf Run, in addition to other Lexington streams, led the US
Environmental Protection Agency (USEPA) and the Kentucky Environmental and Public
Protection Cabinet (KY EPPC) to file a lawsuit against Lexington in 2006 for violations
of the Clean Water Act in 2006. The lawsuit was due to failure of the city to maintain the
sanitary and storm sewer systems, which caused raw sewage discharges into streams. On
March 14, 2008 LFUCG entered into a Consent Decree in order to resolve this lawsuit
(United States, 2006). Within the Consent Decree, LFUCG agreed to make extensive
improvements to its sewer systems and address sanitary sewer overflows and associated
MS4 permit violations, as well as to reduce the discharge of pollutants via stormwater.
With the Consent Decree in place, LFUCG is furthering its efforts to improve water
quality in Wolf Run.
________________________________________________________________________
Wolf Run Watershed
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The citizens of Lexington, especially those in the Wolf Run watershed, share the interest
in water quality improvement with LFUCG. The Friends of Wolf Run (FOWR), a
community based watershed group, became active in the watershed in 1997, prior to the
first impaired listing of Wolf Run, educating the community about stream health and
making initial steps toward a cleaner watershed. This group continues to be an outspoken
proponent of improving the water quality in Wolf Run. The FOWR sponsors the Wolf
Run Watershed Council, consisting of groups and individuals working to improve the
watershed.
A watershed plan is being developed in order to provide a comprehensive assessment of
the health of the watershed, citizen and stakeholder concerns, watershed remediation
strategies, and implementation plans for the future. This is being developed under a
Section 319(h) Nonpoint Source Implementation Program Cooperative Agreement
(#C9994861-09) awarded by the Commonwealth of Kentucky, Energy and Environment
Cabinet, Department for Environmental Protection, Division of Water (KDOW) to
LFUCG based on an approved work plan. These federal funds were awarded to KDOW
by the EPA under Section 319(h) of the Clean Water Act. Third Rock was selected as the
environmental consultant for work under this grant through a request for proposal issued
by LFUCG. FOWR was also issued grant funding through a memorandum of agreement
with LFUCG, primarily to engage, educate, and solicit input from the public during the
development of this plan.
In the development of the Wolf Run Watershed Based Plan (WBP), all known and
relevant existing information pertaining to the watershed was compiled and evaluated for
data quality. The purpose of the data compilation and assessment was to thoroughly
describe the Wolf Run watershed and to determine what additional data would be
necessary in order to identify the impairments in the watershed and their causes and
sources, to calculate the extent of the impairments, and to determine solutions for
improving water quality. Based on this analysis, six major sampling needs were
identified, which include:
 measurements to characterize of the discharge hydrograph for the Preston Springs
karst basin
 watershed conductivity survey
 macroinvertebrate collections on tributaries and headwaters
 watershed-wide habitat assessments
 hydrogeomorphic assessment of the watershed
 a water quality monitoring data set meeting the specifications of KDOW’s
“Watershed Planning Guidebook for Kentucky Communities” (KWA and
KDOW, 2010)
This QAPP will establish the quality criteria and collection process necessary to produce
data which will fill the identified gaps and allow for the determination of the locations in
the watershed in which BMPs will be most feasible, efficient, and effective.
________________________________________________________________________
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A6. Project/Task Description
A6.1. General Overview of Project
This project will involve six different monitoring activities, as follows:
 karst hydrograph characterization
 conductivity survey
 benthic macroinvertebrate collection
 watershed habitat assessment
 hydrogeomorphic assessment
 water quality monitoring
The sampling responsibilities, frequencies and number of sites to be sampled for each
monitoring activity are summarized in Table 2. The equipment necessary for each of
these activities is specified in Table 3. Exhibits 1 and 2 show the study area and the
selected sampling locations. A discussion of each of these activities follows.
TABLE 2 – MONITORING ACTIVITY OVERVIEW
Sampling
Responsibility
Third Rock
Volunteers
Frequency
Twice
Once
No. of Sites
6
1/ 100 ft
Third Rock
Third Rock and
Volunteers
Once
6
Once
6
Watershed Habitat Assessments
Volunteers
Once
Hydrogeomorphic Assessment
Water Quality Monitoring
Third Rock
Twice
Third Rock and
Volunteers
Third Rock and
Volunteers
Monthly,
10 events
5 events in
30 days
Monitoring Activity
Karst hydrograph characterization
Conductivity survey
Benthic Macroinvertebrate Collection
a. Macroinvertebrate Collection
b. Habitat Assessment
a. Water Quality Monitoring
b. E. coli Geomean Sampling
At least 1 per
Segment (24)
9
12
12
________________________________________________________________________
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ow
n
n
Tow
Br
Ol
d
(
!
(
rg
Geo
Le
e
wn
To
´
Tow n
Branprovided by LFUCG, Oct. 2010.
Mapping
ch
Karst mapping from KY Geological Survey.
TABLE 3 – SAMPLING EQUIPMENT SUMMARY
Monitoring Activity
Karst hydrograph
characterization
Conductivity survey
Benthic Macroinvertebrate
Collection
Habitat Assessment
Hydrogeomorphic
Assessment
Water Quality Monitoring
and E. coli Geomean
Sampling
Equipment
Marsh McBirney Flo-Mate Portable 5 Level TROLL
Flowmeter
1 Baro TROLL
Stopwatch
RuggedReader and communication
Tape measure (100 feet in 1/10ft
cables
increments)
PVC pipe, elbow, and caps
Top-setting wading rod
Steel cable
Five gallon bucket
Lap top computer
Field Notebook
Pliers
Wrench
Hammer
Metal fasteners
Steel fence post
Digital Camera
EC Conductivity PockeTesters, or
Field Datasheet
equivalent
GPS
Medium-sized bucket
600μm mesh, 0.25 meter wide 300μm nitex sampler/mesh
rectangular net or kick seine
Fine-tipped forceps
800 x 900μm D-frame dip net
95% ethyl alcohol
U.S. Number 10 sieve
White picking pans
Sample jars
U.S. Number 30 sieve
2- 600μm mesh wash buckets
Digital Camera
GPS
High-Gradient Habitat Assessment
Site Characterization Form
Field Data Sheet
Gravelometer
0.75-inch rebar
GPS
Hammer
Laser level & tripod
Tape measure
100ft & 50ft tapes
Surveying rod
RiverMorph on Rugged Reader
Field notebook
Flow measurement
Field Measurements
Marsh McBirney Flo-Mate Portable
EC Conductivity PockeTesters
Flowmeter, or equivalent
Dissolved Oxygen Water Quality
Stopwatch
Test Kit (LaMotte Code 7414)
Tape measure (100 feet in 1/10ft
2 pH Wide Range Indicator Kits
increments)
(LaMotte P-5085 and P-5100)
Top-setting wading rod
Armored Thermometers (-5° to
Five gallon bucket
45°C in 0.5°C increments)
Field Notebook
All Sampling
Field Filtration
Sample coolers
47mm magnetic filter funnel
Ice
1L Nalgene flask
Plastic food storage bags
Teflon or Tygon tubing
Sample jars and preservatives
Forceps
Powderless latex or nitrile gloves
0.45μm sterile membrane filters
Chain-of-custody
Deionized water
Permanent marker
Clear masking tape
Blue or black ink pen
________________________________________________________________________
Wolf Run Watershed
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A6.1.1. Karst Hydrograph Characterization
To determine the influence of the karst system on the discharge and the nature of the
stream hydrograph, simultaneous gaging of the three affected tributaries and a major
sinkhole will be performed during base flow conditions and during a wet weather event.
Temporary water level gages (pressure transducers with data loggers) will be installed at
each of the five gaging stations.
Flow measurements will be conducted according to KDOW’s Measuring Stream
Discharge Standard Operating Procedure (KDOW 2010b). A minimum of 5 percent
replicate measurements will be made during this gaging effort. The base flow event will
consist of a single flow measurement at each of six gaging stations as shown on the
attached Exhibit 2. It is anticipated that the base flow period will begin in mid-summer
and extend to early fall. The wet weather event will target a storm event that is expected
to have uniform rainfall across the watershed with expected accumulation of over 1 inch.
The gaging will be performed by two teams of surveyors circulating to each of the five
gaging points a minimum of every thirty minutes during the storm event. Monitoring
will continue until well past the hydrograph peak.
A6.1.2. Conductivity Survey
During medium to low-flow conditions (0.5 to 5 cfs at the USGS gage), the Wolf Run
watershed will be surveyed by in situ field temperature and specific conductance
measurements to identify locations of “jumps” in the specific conductance levels as
possible locations of pollution. Using GPS data loggers, field meters, data sheets, and
photographs, all streams and tributaries (approximately 13.5 miles) will be measured at
approximately 100-foot intervals (approximately 700 locations). Volunteer samplers will
be trained to perform this study. Unless interrupted by a precipitation event, the survey
should be completed over a one-week period. If a precipitation event occurs during this
period, the survey will resume when water levels are consistent with the initial survey
conditions.
A6.1.3. Benthic Macroinvertebrate Collection
Macroinvertebrate samples will be collected at six sites within the Wolf Run watershed in
addition to the site at the mouth of Wolf Run, which is monitored for macroinvertebrates
under the MS4 permit. The six sites are located on Vaughn’s Branch, Big Elm Tributary,
Cardinal Run, McConnell Branch, and two sites on Wolf Run (one upstream of
Harrodsburg Road, one upstream of Versailles Road). These sites are identified on
Exhibit 1.
The macroinvertebrate community at each site will be sampled using the recommended
methods developed by KDOW (2009b, 2009c), which involve the collection of two
separate samples, riffle and multihabitat. The riffle sample consists of four 0.25 meters2
(m2) samples collected from two separate riffles at each station using a 0.25 m2 grid and a
kicknet (600μm mesh). Riffle collections at each station will be composited to form one
semi-quantitative sample. The qualitative, multihabitat sample includes, where habitat is
available, samples from leaf packs; sticks/wood; bedrock/slabrock; undercut
banks/submerged roots; aquatic macrophyte beds; soft sediment (using a U.S. # 10 sieve);
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hand-picking of rocks (large cobble/small boulder) from riffles, runs, and pools;
aufwuchs material off rocks, sticks, leaves, and filamentous algae; and visual searches of
large woody debris. All samples collected with the dip net and the rock and wood
samples will be processed through a 600μm wash bucket. Results of qualitative sampling
from each microhabitat will be combined to form one composite sample for each station.
Samples will be preserved in 95 percent ethanol and returned to the laboratory for
processing and identification. All organisms will be identified to the lowest possible
taxonomic level and recorded on laboratory data sheets. Random 300-specimen
subsamples will be removed from the riffle samples using methods described by KDOW
(2009b).
Habitat assessments will be performed by trained volunteers accompanied by Third Rock
personnel at each of these sites. Visual assessments will be made to document riffle and
pool substrates, stream channelization, riparian conditions and in-stream cover. Habitat
assessment procedures will follow those outlined in Rapid Bioassessment Protocols for
Use in Wadeable Streams and Rivers (Barbour et al. 1999).
A6.1.4. Watershed Habitat Assessments
In addition to the habitat assessments conducted at the macroinvertebrate sites, habitat
assessments will be performed by trained volunteers throughout the watershed on parcel
sized or 100m stream reaches. Using the visual-based habitat assessment procedures in
Barbour et al. 1999, volunteers will survey as many segments as time permits within the
24 stream segments into which the watershed has been subdivided. At least one
assessment will occur in each segment.
Information obtained from the habitat assessment will be used to supplement biological
and physicochemical data when determining the overall health of the stream reach and
stream-use designation. Additionally, habitat assessments will serve as a baseline to
document physical changes that occur over time and to identify potential areas for BMP
implementation.
A6.1.5. Hydrogeomorphic Assessment
Nine hydrogeomorphic monitoring sites have been preliminarily designated to measure
channel changes in representative reaches. Assessment will include a series of spatially
integrated, high-resolution cross-section and longitudinal profile surveys and streambed
substrate evaluation to determine the extent of the effects of hydromodification in the
Wolf Run watershed. Effects of hydromodification that may be revealed by the
assessment include degree of bed and bank erosion, sedimentation, and habitat loss. The
relative potential for improvement will also be qualitatively assessed based on the lack of
obvious physical constraints in a reach, position in the landscape, or position in the
watershed.
The baseline cross-section, profile and bed substrate will be compared to a subsequent
survey to determine the degree and type of changes in physical structure and stream
function that has occurred. Information obtained from the hydrogeomorphic assessment
will be used to supplement biological, physicochemical, and habitat data when
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Wolf Run Watershed
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determining the overall health of the stream reach and stream-use designation. This will
also allow for the development of an understanding of the nature and location of the
problems associated with channel modification. Hydrogeomorphic assessments will
quantify physical stream changes that occur over time, help identify potential
BMPs/implementation solutions, and prioritize reaches for implementation of those
solutions.
A6.1.6. Water Quality Monitoring
Water quality monitoring will be conducted at ten (10) monthly sampling events at a
minimum of twelve (12) sampling stations in the watershed during dry and wet
conditions. The sampling date within each month will be flexible such that at least two
of the events are considered ‘wet-weather’ and two of the events are considered ‘dryweather.’ Sampling parameters will include discharge, E. coli, fecal coliform, total
suspended solids, total phosphorus, ortho-phosphorus, ammonia, total kjeldahl nitrogen,
nitrate, nitrite, total dissolved solids, turbidity, dissolved oxygen, specific conductance,
temperature, and pH. The LFUCG Town Branch laboratory will analyze samples for E.
coli, fecal coliform, total suspended solids, ammonia, nitrite, total dissolved solids,
alkalinity, and hardness. The Kentucky Geological Survey will analyze samples for total
phosphorus, ortho-phosphorus, total kjeldahl nitrogen, and nitrate. FOWR volunteer
samplers will perform field measurements of turbidity, dissolved oxygen, specific
conductance, temperature, and pH. Third Rock will accompany the volunteers during
each event to conduct discharge monitoring and will field filter ortho-phosphorus
samples. Also due to the short time period in which wet-weather events can be collected
on the hydrographic rise, two wet-weather sampling events will be collected solely by
Third Rock staff. If for some reason volunteers are not able to perform the sampling,
Third Rock will collect all sampling parameters.
In addition to the monthly sampling, volunteers will collect an additional four events
within a 30-day period during the Primary Contact Recreation period (May 1 to
October 31) for E. coli and fecal coliform to evaluate the geometric mean for the primary
contact period. A Third Rock staff member will accompany the volunteers during each
event to conduct discharge monitoring. Only flow, E. coli and fecal coliform will be
collected during these events. The LFUCG Town Branch laboratory will analyze the
samples.
A6.2. Project Timetable
The project schedule for each of the monitoring activities as well as the data analysis,
report completion, and watershed based plan section revisions are shown in Table 4.
The QAPP completion and approval is expected to occur in April 2011, prior to the initial
volunteer training. If the QAPP approval is delayed beyond April, the monitoring
schedule will be delayed a similar period. The first volunteer training event is scheduled
for April 16, 2011, with four total training sessions planned. FOWR and its Science
Advisors are to conduct these training sessions in accordance with this QAPP.
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Wolf Run Watershed
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The Wolf Run Watershed Council will meet on a quarterly basis, at minimum. Progress
reports on the monitoring activities will be presented at these meetings.
For the karst hydrograph characterization, data loggers will be installed in May 2011.
Subsequent to their installation, the base flow event and wet weather event will be
monitored during the respective weather conditions. A rainfall event of 1 inch will
qualify as a wet weather event. Both events are expected to occur by the end of August
2011 if weather conditions allow. Data analysis will follow the events with the expected
completion data of the report in May 2012.
Trained volunteer samplers will conduct the conductivity survey during medium to lowflow conditions (0.5 to 5 cfs at the USGS gage). Unless interrupted by a precipitation
event, the survey will be completed over a one-week period such that the results reflect a
“snapshot” of watershed conditions. If a precipitation event occurs during this period, the
survey will resume when water levels are consistent with the initial survey conditions.
The sampling will be performed during a period between May and September 2011. All
hardcopy and electronic data will be submitted to Third Rock for data entry and analysis,
with a final report expected by December 31, 2011.
Third Rock will sample macroinvertebrates in May 2011 during the appropriate sampling
index period. For wadeable streams with a watershed >5 mi2, the index period is May 1
to September 30; for headwater streams with a watershed <5 mi2, it is February 15 to
May 31.
All sites are headwater sites except W1, which is sampled for
macroinvertebrates under the MS4 permit and not under this project. Sampling will not
occur during periods of excessively high or low flow or within two weeks of a known
scouring flow event. Habitat assessments will be performed by trained volunteers
concurrent with or within one week of the macroinvertebrate collections at each of these
sites. Laboratory identification, metric calculation, data analysis, and report completion
will occur prior to December 31, 2011.
The watershed habitat assessments performed within the 24 stream segments in the Wolf
Run watershed will be performed subsequent to volunteer training and prior to October
31. All data will be compiled into an electronic database by the volunteer sampling
coordinator and submitted to Third Rock by October 31, 2011.
Hydrogeomorphic monitoring will initially be conducted in May 2011 with a second
survey to assess geomorphic conditions nine (9) months after the baseline is completed
(February 2012), assuming sufficient flow events occur during this period. Data analysis
will occur subsequent to the data generation, with a final report completed by April 30,
2012.
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Wolf Run Watershed
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TABLE 4 – PROJECT SCHEDULE
Activity
Apr
2012
2011
May Jun Jul Aug Sep Oct Nov Dec Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec
Planning and Training
QAPP Approval
Volunteer Training
Council Meetings
Quarterly
nd
rd
Installation of Data loggers
Base Flow Event
Wet Weather Event
Data Analysis
Report Completion
Conductivity Survey
th
3
2
Karst Hydrograph Characterization
th
4
th
5
6
th
7
th
8
th
9
Rainfall dependent
Complete within one week
Conduct survey
Data Entry and Analysis
Report Completion
Benthic Macroinvertebrate Collection
Macroinvertebrate Collection
Headwater
Index
Macro sites
Habitat Assessment
Laboratory identification
Data Entry and Analysis
Report Completion
Watershed Habitat Assessment
Habitat Assessments by Reach
Data Entry and Analysis
Report Completion
Hydrogeomorphic Assessments
Cross-sections, profiles, and
pebble counts
Data Analysis
Report Completion
Water Quality Monitoring
Monthly Sampling (10)
E. coli Geomean Sampling
Data Entry and Review
Loading Calculations and
Source Determinations
Report Completion
Watershed Based Plan
Initial
1
Final
2
3
4
5
6
7
8
9
10
4X in 30 days
Chapter 3: Monitoring
Chapter 4: Analysis
Chapter 5: BMPs
Chapter 6: Strategy
Chapter 7: Implementation
Water quality monitoring will be conducted monthly beginning in May and continuing
until February 2012. At least two of the events are to be considered ‘wet-weather’ and
two of the events are considered ‘dry-weather.’ Representative dry weather sampling
conditions are defined by an antecedent period of 5 days during which there is no rainfall.
Representative wet weather sampling conditions are defined by a period of at least 72
hours of dry weather preceding a rain event with a total accumulation of at least 0.1 inch.
Sampling during ‘wet-weather’ will occur on the hydrographic rise. Additional sampling
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for E. coli will occur four times within 30 days and is expected to begin in May, but may
occur at any time within the Primary Contact Recreation period (May 1 to October 31).
Third Rock will accompany volunteers during each event to conduct discharge
monitoring. A minimum notice of 48 hours will be provided to FOWR prior to
scheduling the volunteer sampling. The LFUCG Town Branch and Kentucky Geological
Survey laboratories will analyze collected samples. Collection events shall be scheduled
to avoid collection on Thursday, as this day is particularly busy for the Town Branch
Laboratory. Expected turn-around-time for the laboratory analysis is 30 days. Chemical
laboratory reports with data quality review by the Project QA Manager will be submitted
to the project team within 60 days of sample delivery to the laboratory. With each
sampling event, quality control samples including field duplicate samples will be
collected along with the regular field samples.
The results of these studies will be used to complete the Wolf Run Watershed Based Plan
according to the KDOW’s “Watershed Planning Guidebook for Kentucky Communities”
(2010) and the USEPA’s Handbook for Developing Watershed Plans to Restore and
Protect Our Waters (2008). The project schedule in Table 4 corresponds to the
watershed plan chapters outlined in the KDOW guidance. The final plan, incorporating
comments and recommendations, is scheduled for completion by December 31, 2012.
A7. Data Quality Objectives (DQOs) and Criteria for Measurement Data
A7.1. Data Quality Objectives
Data quality objectives (DQOs) are qualitative and quantitative statements that clarify the
intended use of the data, define the type of data needed to support the decision, identify
the conditions under which the data should be collected, and specify tolerable limits on
the probability of making a decision error due to uncertainty in the data. The data quality
objectives for the respective sampling activities are listed in Table 5, along with the
Standard Operating Procedures associated with each of these activities. The overall
objective of this project is to collect data of sufficient quality and quantity to support the
development of a watershed-based plan for the Wolf Run watershed.
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Wolf Run Watershed
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TABLE 5 – SUMMARY OF DATA QUALITY OBJECTIVES AND STANDARD
OPERATING PROCEDURES
Sampling Activity
Karst Hydrograph
Characterization
Conductivity Survey
Objective
Characterize
the
flow
distribution
between
the
surface water and ground water
systems.
Identify significant changes in
conductivity levels to pinpoint
sources of pollution
Macroinvertebrate
Collection and
Identification
Calculation
of
the
Macroinvertebrate
Bioassessment Index (MBI).
Macroinvertebrates
have
varying tolerances for water
pollution and therefore can
serve as long-term indicators
of water quality
Habitat Assessment
Provide a semi-quantified
evaluation of the general
habitat of the stream
Hydrogeomorphic
Sampling
Determine the degree to which
streams are being effected by
hydromodification including
erosion rates, changes in the
streambed, and position within
the cycle of channel evolution
in order to guide and prioritize
remediation efforts
Grab sampling
Identify loading of water
quality parameters to identify
whether specific pollutants are
causing impairments in the
watershed
Standard Operating Procedures / References
KDOW. 2010b. Measuring Stream Discharge Standard Operating Procedure.
Kentucky Department for Environmental Protection, Division of Water,
Frankfort, Kentucky. DOWSOP03019
In-Situ Inc. 2006. Level TROLL® Operator’s Manual. www.in-situ.com
Oakton Instruments. Waterproof TDSTestr and ECTestr Series Instructions.
http://www.4oakton.com/Manuals/ConductivityTDS/TDS_ECTestrmnl.pdf
KDOW. 2009b. Laboratory Procedures for Macroinvertebrate Processing and
Taxonomic Identification and Reporting.
Kentucky Department of
Environmental Protection, Division of Water, Frankfort, Kentucky.
KDOW. 2009c. Methods for Sampling Benthic Macroinvertebrate Communities
in Wadeable Waters. Kentucky Department for Environmental Protection,
Division of Water, Frankfort, Kentucky.
KDOW. 2008. Methods for Assessing Biological Integrity of Surface Waters in
Kentucky. Kentucky Department of Environmental Protection, Division of
Water, Frankfort, Kentucky.
Barbour, M.T., J. Gerritsen, B.D. Snyder, and J.B. Stribling. 1999. Rapid
Bioassessment Protocols for Use in Streams and Wadeable Rivers:
Periphyton, Benthic Macroinvertebrates and Fish. Second Edition. EPA
841-B-99-002. USEPA, Office of Water, Washington, D.C.
Bunte, Kristin; Abt, Steven R. 2001. Sampling surface and subsurface particlesize distributions in wadable gravel-and cobble-bed streams for analyses in
sediment transport, hydraulics, and streambed monitoring. Gen. Tech. Rep.
RMRS-GTR-74. Fort Collins,CO: U.S. Department of Agriculture, Forest
Service, Rocky Mountain Research Station. 428 p.
Harrelson, C.C., C.L. Rawlins, and J.P. Potyondy. 1994. Stream channel
reference sites: An illustrated guide to field technique. General Technical
Report RM-245. Fort Collins, CO: U.S. Department of Agriculture, Forest
Service, Rocky Mountain Forest and Range Experiment Station. 61p.
Rosgen, D.L. 2008. River Stability Field Guide. Wildland Hydrology, Pagosa
Springs, CO.
KDOW. Watershed Watch Water Chemistry Sampling Methods for Field
Chemistry and Lab Analysis. http://www.lrww.org/training/chem-test.pdf
KDOW. 2009a. In-situ Water Quality Measurements and Meter Calibration
Standard Operating Procedure. Kentucky Department for Environmental
Protection, Division of Water, Frankfort, Kentucky. DOWSOP03014
KDOW. 2010b. Measuring Stream Discharge Standard Operating Procedure.
Kentucky Department for Environmental Protection, Division of Water,
Frankfort, Kentucky. DOWSOP03019
KDOW. 2009b. Sampling the Surface Water Quality in Lotic Systems. Kentucky
Department for Environmental Protection, Division of Water, Frankfort,
Kentucky. DOWSOP03015
LaMotte. Dissolved Oxygen Water Quality Test Kit Instruction Manual. Code
7414 / 5860.
LaMotte Company.
Chestertown, Maryland.
www.lamotte.com
Price, David J. 2009. Quality Assurance Plan (QAP) and Standard Operating
Procedures (SOPs). Lexington-Fayette Urban County Government Division
of Water Quality Town Branch Laboratory.
Kentucky Geological Survey Standard Operating Procedures. See Appendix B.
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A7.2. Data Quality Indicators
When measurement performance or acceptance criteria can be stated in quantitative
terms, they are called data quality indicators (DQI). The quality of field and analytical
data is most often assessed in terms of the DQIs including precision, accuracy/bias,
representativeness, comparability, completeness, and sensitivity.
A7.2.1. Precision
Precision is the measure of agreement among repeated measurements of the same
property under identical or near identical conditions. It is usually calculated as a range,
standard deviation, or relative percent difference (RPD). Relative percent difference will
be the primary measure of precision for laboratory and field duplicates on this project,
and is calculated as follows:
RPD%  
SD
 100
S D


 2 
where:
S = first sample value (original result)
D = second sample value (duplicate result)
The project standards for precision on in-situ measurements and water quality grab
samples are expressed in Table 5. The precision of in-situ measurements will be assessed
by one duplicate measurement during each sampling event. The precision of the water
quality grab samples will be measured by internal laboratory QC samples. In addition to
internal laboratory QC, one field duplicate will be collected per sampling event. A field
duplicate or field replicate sample is a sample taken from the same location as the
‘regular’ grab sample, at the same time. The sample is used to assess variability of
environmental conditions at sampling sites.
For flow measurement, replicate measurements will be made to test the accuracy of the
individual making the measurements. Replicate measurements are made by the same
individual who made the original measurements and at the same cross section as the
original, but with different horizontal locations (stations) across that cross-section. For
example, if the original cross section had stations at even intervals (2, 4, 6, 8 etc.), the
replicate measurement might have stations set at odd intervals (3, 5, 7, 9 etc.).
For benthic macroinvertebrate identification, ten percent of all sorting pans will be
randomly checked by a second sorter to assure that samples have been picked thoroughly.
Five percent of all identified samples will randomly be re-identified to insure QA/QC by
a second taxonomist. Ninety percent or greater composition comparability (e.g.,
abundance and richness) is the target success criteria. If there is less than 90 percent
comparability between the taxonomists, then taxonomy must be reconciled by both
taxonomists and a third taxonomist, if deemed necessary.
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Wolf Run Watershed
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TABLE 6 – ACCEPTANCE CRITERIA FOR WATER CHEMISTRY AND INSITU MEASUREMENTS
Parameter
Units
Method
Accuracy
(%R or ±)
Precision*
(% RPD)
Dissolved Oxygen
Specific Conductance
pH
Water Temperature
Turbidity
Flow
Total Dissolved Solids
Total Suspended Solids
Total Alkalinity
Hardness
Escherichia coli
Fecal Coliform
Ortho-phosphorous as P
Phosphorus, Total as P
Ammonia as N
Nitrate as N
Nitrite as N
Total Kjeldahl Nitrogen as N
mg/L
μS/cm
SU
˚C
Visual
cfs
mg/L
mg/L
mg/L CaCO3
mg/L CaCO3
MPN/100mLs
CFU/100mLs
mg/L PO4-P
mg/L PO4-P
mg/L NH3-N
mg/L NO3-N
mg/L NO2-N
mg/L TKN-N
LaMotte
Oakton
Watershed Watch
Watershed Watch
Visual Observation
DOWSOP03019
EPA 160.1
SM 2540 D
SM 2320 B
SM 2340 C
SM 9221 E
SM 9221 F
EPA 365.1
EPA 365.2
EPA 350.1
EPA 300.0
SM 4500-NO2 B
SM 4500-Norg C
±1.5
95-105
±0.5
±0.5
NA
±0.05 ft/sec
95-105
95-105
80-120
80-120
N/A
N/A
80-120
80-120
80-120
80-120
80-120
80-120
20
20
20
20
NA
N/A
20
20
20
20
20
20
20
20
20
20
20
20
Sensitivity
(Reporting
Limit)
0.5
10
NA
-5 to 45
NA
0.01 ft/sec
10
2
0
0
1
1
0.05
0.02
0.05
0.02
0.02
0.5
* Indicates minimum laboratory precision for all parameters except in-situ measurements. For in-situ, this indicates field precision.
For hydrogeomorphic sampling, the surveying precision of cross-sections and profiles
shall be +/- 0.01 ft for vertical readings and +/- 0.1 ft for horizontal readings. The laser
level precision shall be less than +/- 3.0 mm/30m. Precision for pebble count readings
will be such that each data point measures within +/- 1 unit of the narrative particle
description or +/- 0.5 phi units on the gravelometer.
A7.2.2. Accuracy
Accuracy is a measure of overall agreement between a measurement and a known value.
Accuracy includes an evaluation of bias, which is a systematic or persistent distortion of
a measurement process that causes errors in one direction. Accuracy is quantified by
calculating the percent recovery (%R) of a known quantity of an analyte under a particular
test method as follows:
%R 
Vm
 100
Vt
where:
Vm = measured value (determined by analysis)
Vt = true value (as calculated or certified by a manufacturer)
No water quality field samples will be collected in order to evaluate accuracy. However,
internal laboratory QC samples will be analyzed to determine if the project standards,
listed in Table 4, are met. For benthic macroinvertebrate samples vouchers are collected
________________________________________________________________________
Wolf Run Watershed
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to ensure accuracy.
assessment.
Field photographs are used to document accuracy for habitat
For the conductivity survey, the GPS units used to document locations shall be accurate
to at least 20 feet, and the latitude and longitude shall be recorded in decimal degrees to
six decimal places in datum NAD83.
A7.2.3. Representativeness
Representativeness is the degree to which data accurately and precisely represent an
environmental condition. Representativeness is largely a product of proper selection of
sampling sites within the watershed and proper execution of the methodology. For
instance, per the sampling method, grab samples are to be collected from the thalweg and
not from the bank in order to ensure representativeness. Representativeness is also
ensured by collection under the specified sampling conditions. Representative dry
weather sampling conditions are defined by an antecedent period of 5 days during which
there is no rainfall. Representative wet weather sampling conditions are defined by a
period of at least 72 hours of dry weather preceding a rain event with a total accumulation
of at least 0.1 inch. Sampling during ‘wet-weather’ will occur on the hydrographic rise.
Representative conditions for macroinvertebrate sampling are established by the
respective index periods for sampling. In addition, macroinvertebrate samples will not be
collected during periods of excessively high or low flows or within two weeks of a
known scouring flow event.
Other anomalous conditions or unusual land uses at the time of sampling will be recorded
in the field notebook.
A7.2.4. Completeness
Completeness is a measure of the amount of valid data needed to be obtained from a
measurement system. The completeness goals for each of the sampling types are
summarized in Table 7. Dry weather is expected to produce no flow conditions at several
sites during the sampling period.
TABLE 7 – COMPLETENESS GOALS OF SAMPLING ACTIVITIES
Sampling Activity
Karst Hydrograph Characterization
Conductivity Survey
Macroinvertebrate Collection
Habitat Assessment
Hydrogeomorphic Sampling
Grab sampling
Completeness Goal
At each site, a dry-weather event and measurements from the firstflush to past the peak flow during a wet-weather event
Minimum of 500 sites with at least one in each of the 24 stream
segments
All six sites
Minimum of one site from each of the 24 stream segments
All nine sites – two surveys
Five samples in 30 days for E.coli.
At least two ‘dry-weather’ and two ‘wet-weather’ events at all
sites; no flow conditions are expected during dry ant hot weather
conditions at several headwater sites
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A7.2.5. Comparability
Comparability is a term that expresses the measure of confidence that one data set can be
compared to another and can be combined for the decision to be made. Comparability
may be assessed by comparing sampling methodology, analytical methodology, and units
of reported data. The standards of quality established in this QAPP are consistent with
the previously collected data in the Wolf Run watershed collected under separate QAPPs.
All data to be utilized in the generation of the watershed based plan source and loading
determinations will have been generated under an approved QAPP.
A7.2.6. Sensitivity
Sensitivity is the capability of a method to discriminate between measurement responses
representing different levels of the variable of interest. Sensitivity is particularly
important for ensuring that the measurement levels are sufficient to detect whether
particular pollutants are present at levels that may cause impairment to the designated
use. For grab sampling and in-situ measurements, the sensitivity levels necessary for this
program are specified in Table 6. For macroinvertebrate sampling, all organisms are to be
identified to the lowest possible taxonomic level possible in order to properly calculate
the associated metrics.
A8. Training Requirements
All volunteers involved in the sampling for this project shall have successfully completed
the training workshops lead by a trainer registered with the KDOW Watershed Watch
Volunteer Database as administered by the Watershed Management Branch. The training
will consist of four sessions that will cover habitat assessments, grab sample collection,
in-situ measurements, and the requirements of this QAPP. Powerpoint presentations to
be used in this training are available at http://www.lrww.org/training/. The training will
be organized and conducted by the FOWR and their Science Advisors. The FOWR
Sampling Coordinator will maintain documentation of the volunteer sampler training
through the Participant Agreement Form (Appendix A).
A9. Documentation and Records
In order to provide quality data that meets the project objectives, traceability and
maintenance of documentation and records is essential. All records relating to the
collection, analysis, or reporting of data associated with the project shall be made
available upon request by KDOW or LFUCG. A summary of such documentation is
included below.
A9.1. Field Documentation and Records
Field records will include all data recorded in the field including completed field
datasheets, field logbooks, monitoring records, and chain of custody sheets. All data will
be recorded using black or blue indelible ink, and it is recommended that waterproof
paper be used where feasible. Mistakes on field data sheets will be crossed out with one
line (so the information is still discernible), with the initials and date of the person
making the correction. The correct information should then be recorded legibly on
another line, or above or below the original info. If a separate sheet is necessary for new
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information, the original sheet should be attached to the new sheet, and initialed and
dated.
The following field documents shall be used in this project:
 Site Characterization Form
 High Gradient Stream Data Sheet for Habitat Assessment
 Conductivity Survey Field Data Sheet
 Pebble Count Field Data Sheet
 Water Quality Chain-of-Custody
 Macroinvertebrate Chain-of-Custody
 Sample Labels
 Field “Rite in the Rain” Notebook
 Field “Reference Reach” Notebooks
 Field Meter Calibration Logs
Copies of these documents are found in Appendix A, where applicable.
All raw data collected in the field will ultimately be submitted in the chemical or
biological data package. However, all field notes, including the location and frequency
of QC sampling, in situ measurements, and calibration and maintenance logbooks will be
retained for the duration of the grant period.
Where possible, all field in situ measurements will be recorded on the datasheet or chainof-custody. However, if necessary, results or notes may be maintained in a field
notebook. Equipment calibration and maintenance logs will be documented and recorded
per procedure specifications.
A9.2. Laboratory Documentation and Records
Chemical laboratories are required to maintain a current Quality Assurance Manual
documenting all aspects of their quality system including control of Standard Operating
Procedures (SOPs) and datasheets. Documents issued as part of the quality system will
be reviewed and approved by authorized personnel. A master list identifying the current
revision and distribution of documents in the quality system will be used to ensure that
invalid and obsolete documents are not used. Quality system documents will be uniquely
identified by the date of the last revision, issuing authority, and the total number of pages
or a mark indicating the end of the document. The Lexington-Fayette Urban County
Government Division of Water Quality Town Branch Laboratory Quality Assurance Plan
(QAP) and Standard Operating Procedures (SOPs) (Price 2009) and the Kentucky
Geological Survey’s Standard Operating Procedures located in Appendix B will control
the chemical laboratory document control.
Laboratories will be required to document the analysis of all quality controls associated
with the analysis of the collected samples such that the entire data package, along with a
narrative description of the results and a list of all data qualifiers, may be provided to the
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KDOW and LFUCG upon request. Thus, the laboratories will retain all data associated
with the sample analysis for the duration of the grant period.
Third Rock’s macroinvertebrate identification laboratory will follow laboratory protocols
for benthic macroinvertebrate sample processing, identification and data reporting per
KDOW (2009b, 2008) with the following exceptions:
 All samples will be logged into Third Rock's Macroinvertebrate Laboratory
Information Management System (MacLIMS) upon receipt.
 Sample identification date will be maintained in MacLIMS.
 Taxonomic QA/QC dates (if applicable) will be noted on individual QA/QC
forms and maintained electronically in the Project File.
 Initials of the applicable party completing each task associated with sorting,
identification, or quality control will be noted electronically in MacLIMS or on
associated QA/QC forms.
 QA checks will be documented on applicable forms and maintained in associated
project files. These forms include the Macroinvertebrate Sample Sorting
Efficiency Form, Macroinvertebrate Sample Taxonomy Precision Form, and
Macroinvertebrate Sample Taxonomic and Enumeration Efficiency Form
(Appendix A).
Completed chain(s)-of-custody and sample labels will also be controlled throughout the
analysis process, and completed chain(s)-of-custody will be submitted in the data
package. The laboratory will retain all data associated with the sample analysis for the
duration of the grant period.
Expected turn-around time for the laboratory analysis is 30 days. Chemical laboratory
reports with data quality review by the Project QA Manager will be submitted to the
project team within 60 days of sample delivery to the laboratory. The chemical
laboratory data package will include the laboratory results, completed chain(s)-ofcustody, lists of qualifiers associated with the data, and a report of the quality control
results.
The macroinvertebrate report data package will include a list of the identified species,
metric calculations, habitat assessment scores, photographs, completed chain(s)-ofcustody, and a data analysis report. This report will be submitted to KDOW and LFUCG
prior to December 31, 2011.
A9.3. Quality Documentation and Final Reports
The most recent version of this QAPP will be distributed to all parties listed on the
distribution list after the QAPP has been reviewed and approved. The QA Manager is
responsible for ensuring that all applicable parties perform documented reviews of the
QAPP. If, because of deviations in the QAPP, revisions are required, the QA manager
shall ensure that all parties review the revised version. The current revision and the date
of the revision shall be documented in the upper right corner of the QAPP pages. The
QAPP shall be redistributed after all parties have reviewed the document.
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As a result of this project, multiple final reports will be used to document the findings of
the monitoring performed under this QAPP. Monitoring reports will be submitted to the
KDOW and LFUCG in hardcopy upon request. Electronic data will be presented in
Adobe Acrobat, Microsoft Word, and/or Microsoft Excel depending on the data type.
These final reports are as follows:







Karst Hydrograph Characterization in the Wolf Run Watershed
Assessment of Habitat and Macroinvertebrates in the Wolf Run Watershed
Conductivity Survey of the Wolf Run Watershed
Hydrogeomorphic Assessment of the Wolf Run Watershed
Wolf Run Watershed Monitoring Report
Wolf Run Watershed Based Plan
Final Project Report
The Wolf Run Watershed Monitoring Report will include an evaluation of the quality
assurance and will compare the data produced under the water quality monitoring to the
data quality indicators listed herein. The Wolf Run Watershed Based Plan will be
developed in accordance with the KDOW’s Watershed Planning Guidebook for Kentucky
Communities (2010) and meeting the USEPA’s nine key elements for watershed based
plan. The Final Project Report will meet the requirements of the KDOW Project Final
Report Guidelines for Clean Water Act §319(h)-Funded Projects (2004).
In addition to these reports, quarterly Section 319(h) Nonpoint Source Project Progress
Report will be submitted to the KDOW to document the progress on the project
milestones.
SECTION B. - DATA GENERATION AND ACQUISITION
B1. Sampling Process Design
As previously mentioned, this QAPP addresses six different monitoring types. A
summary of the sampling locations and rationale behind the monitoring types is provided
below.
B1.1. Sampling Site and Reach Locations
Exhibits 1 and 2 indicate the locations of the permanent sampling sites on aerial and
geologic quadrangle mapping. Table 8 describes the location of each sampling site and
whether monitoring for the karst hydrograph characterization, macroinvertebrate
collections, hydrogeomorphic sampling, or water quality monitoring will be performed at
these sites.
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TABLE 8 – SAMPLING LOCATIONS
Site
Name
Stream
Location
Directions
Downstream of Old Frankfort Pike
Old Frankfort
northbound prior to roundabout at
Pike
Alexandria Dr.
Park to the right of the Assembly of God
McConnell Prestons
Church on Dunkirk Drive. Access site
W2
Branch
Cave
through grass lot next to the church to
spring on the left.
About 50 feet downstream of Cambridge
W3 Wolf Run Valley Park Drive Bridge. Park in Cambridge Park
Appartments parking lot.
About 30 feet upstream of confluence with
Vaughn's
Wolf Run. Sample upstream of the
W4
Valley Park
Branch
sanitary sewer manhole cover. Park in
Cambridge Park Appartments parking lot.
Upstream of Devonport Drive, west of
Cardinal
Devonport Alexandria Dr. Park in Pleasant Cove
W5
Run
Dr
Apartments lot to left of bridge, take
sample at rock upstream of bridge.
Park on Cross Keys Road, just off of
Cardinal
Parkers Mill
W5A
Parkers Mill Road. Sample reach as
Run
Rd
diverges from curbside.
W1
W6
W7
W8
W9
W10
W11
W11A
W12
Wolf Run
Take Roanoke Dr off of Alexandria Dr. past
7 Pines Dr. Park uphill of cement slab in
greenway, sample at path crossing creek.
Turn right onto Tazwell Drive off Mason
Pine
Vaughn's
Headly Road. Travel to the end of the
Meadow
Branch
road. Walk below the playground to
Park
sample.
Park at lot to right of Parkway Drive in front
Vaughn's Picadome
of Picadome Golf Course. Site is north of
Branch
Golf Course the clubhouse, midway between the
bridges between holes 4 and 5
At terminus of Faircrest Dr south of Lynn
Faircrest
Wolf Run
Dr. Sample upstream of confluence with
Drive
Springs Branch.
Springs
Faircrest
From W9, cross Wolf Run at foot bridge
Branch
Drive
and sample upstream on Spring Branch.
Park at end of parking lot behind the
Big
Elm Harrodsburg Harrodsburg Rd Fire Station. Access just
Tributary Road
north of power transformer at edge of
adjacent parking lot.
Park at lot behind the Picadome Golf
Big
Elm Picadome
Course clubhouse. Measure discharge at
Tributary Golf Course confluence of surface runoff with Vaughn's
Branch if flow is present.
Lafayette
Sample about 50 feet below bridge at
Wolf Run
Parkway
Rosemont Garden
Wolf Run
Wolf Run
Park
Latitude
Longitude
Monitoring Type
Karst Macro Geomorph WQ
38.067303 -84.554182
X
MS4
X
X
38.057333 -84.542169
X
X
X
X
38.053742 -84.550782
38.054904 -84.549624
X
X
X
X
38.048594 -84.553867
X
X
38.043212 -84.557131
X
X
X
X
X
38.044927 -84.536148
X
X
38.037453 -84.525057
X
X
X
X
38.045274 -84.550661
38.029954 -84.537091
X
X
38.029855 -84.537196
X
38.031245 -84.526027
X
38.037494 -84.527095
O
38.022932 -84.528581
X
X
X
NOTE: “X” indicates permanent sampling location. “O” indicates measurements will occur if flow is present. “MS4” indicates that
sampling is scheduled but under the MS4 permit and not this project.
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Wolf Run Watershed
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X
X
For the conductivity survey and watershed habitat assessments, the watershed has been
divided into twenty-four stream segments as shown in Exhibit 3 and summarized in
Table 9.
TABLE 9 – HABITAT ASSESSMENT STREAM SEGMENTS
Segment ID
1A
1B
1C
1D
1E
1F
1G
1H
1J
2A
2B
3A
3B
3C
3D
3E
3F
4A
4B
4C
5A
5B
6
7
Stream
Wolf Run
Wolf Run
Wolf Run
Wolf Run
Wolf Run
Wolf Run
Wolf Run
Wolf Run
Wolf Run
McConnell Branch
McConnell Branch
Vaughn’s Branch
Vaughn’s Branch
Vaughn’s Branch
Vaughn’s Branch
Vaughn’s Branch
Vaughn’s Branch
Big Elm Tributary
Big Elm Tributary
Big Elm Tributary
Cardinal Run
Cardinal Run
Gardenside Tributary
Unnamed Tributary
Stream Segment
Mouth to Old Frankfort Pike
Old Frankfort Pike to New Circle Rd
New Circle Rd. to Cambridge Dr
Cambridge Dr to Versailles Rd
Versailles Rd to Appomattox Rd
Appomattox Rd to Faircrest Dr
Faircrest Dr to Lafayette Pkwy
Lafayette Pkwy to railroad tracks north of Southland Dr
Railroad tracks to Nicholasville Rd
Wolf Run to Preston's Cave Spring
McConnell Springs Branch through Stormwater Structure
Wolf Run to Oxford Circle
Oxford Circle to Versailles Rd
Versailles Rd to Summerville Rd/Golf Course Fence
Picadome Golf Course
Picadome Golf Course Fence to Gibson Ave Culvert
Simpson Ave to railroad tracks
Sinkhole on Picadome Golf Course to Harrodsburg Rd
Harrodsburg Rd to railroad tracks via Bob-O-Link Dr
Nicholasville Rd to behind Central Baptist Hospital
Wolf Run to Versailles Rd
Versailles Rd to End of Chinquapin Ln
Upstream of Parkers Mill Rd
Wolf Run under Alexandria Dr and Old Frankfort Pike to pond
B1.2. Sampling Design Rationale
B1.2.1. Karst Hydrograph Characterization
The Wolf Run watershed has significant karst development, which must be considered
during loading calculations because it can influence the decision making process during
development of the action plan. In particular, local karst deviation from surface
watershed boundaries adds drainage area to Wolf Run. Based on dye traces, a substantial
fraction of both the Vaughn's Branch and main stem of Wolf Run sub-watersheds are
captured by the Prestons (McConnell) Spring Basin (Recker and Meiman, 1990 and
Spangler, 1992). During base flow and drier conditions most of the surface water in the
karst-influenced fractions of these sub-watersheds is directed to Prestons Spring. During
high flow conditions the surface component of the discharge becomes greater as the karst
system conduit limits are approached. To determine the influence of the karst system,
storm event and base flow gaging of key locations will be conducted to determine the
discharge and the nature of the hydrograph.
________________________________________________________________________
Wolf Run Watershed
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The six gaging locations in the watershed allow for the evaluation of the discharge at the
mouth of the watershed (W1), Prestons Spring (W2), Vaughn’s Branch tributary (W4),
Wolf Run upstream of Cardinal Run (W6), Wolf Run at the edge of the karst basin (W9),
and Big Elm Tributary which flows into the sinkhole at Picadome (W11). In the event
that the capacity of the sinkhole downstream of W11 is exceeded and surface water flows
from Big Elm Tributary into Vaughn’s Branch, this surface flow will be measured at
W11A in order to measure the input into the karst system at this site.
B1.2.2. Conductivity Survey
Specific conductance was recently listed as a cause of impairment in the Wolf Run
watershed. Although specific conductance or conductivity has been analyzed during
several studies and the FOWR conducted a broad study of conductivity levels in the
watershed, a subsequent study under more normal flow conditions will aid in identifying
inputs and problem areas. Conductivity is a measure of water's ability to conduct an
electric current, and it indicates the concentration of dissolved ions in the water. Rapid
changes in the conductivity of a waterbody can indicate groundwater input, catchment
geology, or pollution.
B1.2.3. Benthic Macroinvertebrate Collection
Benthic macroinvertebrate sampling integrates months or even years of water quality
impacts as well as the cumulative effects of multiple stressors and pollutants instead of
particular individual stressors. The KDOW uses biological indicators to determine the
use attainability of a water of the Commonwealth as it relates to KDOW’s narrative water
quality standards. Biological assessment will provide a more accurate evaluation of water
quality health in the watershed. The six macroinvertebrate sites are located on the
tributaries of Wolf Run and in its headwaters to evaluate the macroinvertebrate
communities in the headwaters for comparison to the data collected over multiple years
near the mouth of the watershed.
B1.2.4. Watershed Habitat Assessments
Habitat assessments will be used to supplement biological and physicochemical data
when determining the overall health of the stream reach and stream-use designation.
Additionally, habitat assessments will serve as a baseline to document physical changes
that occur over time and to identify potential areas for BMP implementation. The
watershed was divided into 24 stream segments in order to ensure that assessments are
collected from all representative reaches.
B1.2.5. Hydrogeomorphic Assessment
The process of development within the Wolf Run watershed has affected the stream by
altering watershed hydrology and sediment-transport patterns. The large amount of
impervious surface has greatly reduced the capacity of the watershed to capture and filter
rainfall. Higher runoff rates mean that runoff reaches the stream channels more quickly
(flashier flows) and peak discharge rates are higher compared to an undeveloped
watershed for the same size rainfall event. These effects are known as hydromodification.
Hydromodification can also be direct modification of a stream (for purposes of flood
control, navigation, sediment control, infrastructure protection, etc.), such as
________________________________________________________________________
Wolf Run Watershed
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channelization, armoring, and removal of riparian vegetation. Channel erosion and bank
failure is often caused or exacerbated by hydromodification activities.
Nine hydrogeomorphic monitoring sites have been preliminarily identified throughout the
watershed where quantitative data will be collected to measure channel change in
representative reaches. It is expected that the nine hydrogeomorphic monitoring sites
represent stream reaches that are susceptible to the effects of hydromodification, are in
need of management to stop further degradation, and would be good locations to
implement remediation. Assessment will include a series of spatially integrated, highresolution cross-section and longitudinal profile surveys and streambed substrate
evaluation to determine the extent of the effects of hydromodification in the Wolf Run
watershed. Effects of hydromodification that may be revealed by the assessment include
degree of bed and bank erosion, sedimentation, and habitat loss. The relative potential
for improvement will also be qualitatively assessed based on the lack of obvious physical
constraints in a reach, position in the landscape, or position in the watershed.
B1.2.6. Water Quality Monitoring
The objective of the water quality monitoring is to provide sufficient temporal and
geographic data to evaluate the sources and loadings of water quality pollutants. The
sampling period of ten months was selected in order to evaluate at least one sample from
all seasons. The twelve sampling stations were selected in order to evaluate the relative
contributions of the stream reaches throughout the watershed.
Flow will be measured on the receiving streams because it is a component of the formula
for calculating loading of pollutants in the watershed. Dissolved oxygen, temperature,
and pH will be assessed as basic measurements for describing the health of the stream
and evaluating applicable water quality standards. E. coli and fecal coliform will be used
to assess health risk due to waterborne pathogens. Specific conductance and total
dissolved solids will be used to assess dissolved ions levels present in the watershed.
Turbidity and TSS will be recorded to assess the suspended solid levels for impacts to
stream biota due to increased turbidity, siltation, and other effects. Ortho-phosphorus,
total phosphorus, nitrate, nitrite, and TKN will be assessed to identify imbalances which
may cause eutrophication and impacts to stream biota. Ammonia will be assessed to
evaluate levels for toxicity to plants, animals, and humans. Alkalinity and hardness will
be assessed to measure the buffering capacity of the water against rapid pH changes.
Because E. coli levels will be evaluated against the geomean criteria, it is necessary to
collect five samples in 30 days for this project. For other parameters, at least two ‘dryweather’ and two ‘wet-weather’ events will be sampled at all sites in order to adequately
characterize the loadings geographically. It is expected that no flow conditions will be
observed in the watershed during the sampling period.
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B2. Sampling Methods
B2.1. Sampling Equipment
Equipment to be utilized in sampling is listed in Table 3, Sampling Equipment Summary.
Samples are to be collected and preserved according to the specifications in Table 10.
TABLE 10 – SAMPLE PRESERVATION AND HOLD TIME
Parameter
Analysis Method
Maximum
Holding
Time
E. coli
SM 9221 E
6 hrs
Fecal Coliform
SM 9221 F
6 hrs
Ammonia
EPA 350.1
28 days
Alkalinity
SM 2320 B
SM 2540 D, EPA
160.1, SM 2340 C,
SM 4500-NO2 B
14 days
Ortho-phosphorus
EPA 365.1
28 days
Plastic, 8 oz
Total phosphorus,
TKN
Nitrate
EPA 365.3, SM
4500-Norg C
EPA 300.0
28 days
Plastic, 32 oz
7 days
Plastic, 8 oz
Macroinvertebrate
samples
KDOW 2009c
None
1 Liter wide
mouth plastic
TSS, TDS, Total
Hardness, Nitrite
7 days
Sample
Container
Sample
Preservation
Sterilized
Plastic, 4 oz
Sterilized
Plastic, 4 oz
Cool <6°C,
Na2S2O3 (No Cl2)
Plastic, 8 oz
Cool <6°C,
Na2S2O3 (No Cl2),
H2SO4 to pH <2
Plastic, 32 oz
Ice to <6°C
Deliver To
LFUCG Town
Branch
Laboratory
Field Filter,
Cool <6°C,
H2SO4 to pH <2,
Cool <6°C,
H2SO4 to pH <2
Ice to <6°C
Kentucky
Geological Survey
Laboratory
95% ethanol
Third Rock
Macroinvertebrate
Laboratory
B2.2. Sampling Methods
B2.2.1. Karst Hydrograph Characterization
Because the karst drainage collected by the Prestons Spring Basin emerges as a spring
and surface water, only surface water discharge methods will be used to characterize the
hydrograph of the karst system. Surface water discharge (Q) will be calculated using two
variables, flow area (A) and water velocity (V), according to the equation: Q = AV.
However, because the velocity is variable across a stream cross-section, the flow area and
velocity must be measured in intervals across the stream and summed as shown in
Figure 2. The flow area of each interval is the product of the width (w) and depth (d) for
that interval. The velocity will be measured for each of these areas.
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FIGURE 2 – MEASUREMENT OF DISCHARGE THROUGH SUB-SECTIONAL
MEASUREMENTS
Figure 3– Flow Area
V Calculation
V
d
V
d
d
w
w
V
w
V
d
w
d
w
Note: Stream cross-section showing intervals where water depth and velocity are measured. Flow will be
calculated for each “box” (flow area for each box is d * w) and summed to obtain the flow for the entire stream.
Flow measurements will be conducted according to the KDOW’s Measuring Stream
Discharge Standard Operating Procedure (KDOW 2010b), as described below. Under
this procedure, a tape measure of at least 100 feet is stretched across the stream so that it
is taut and perpendicular to the stream flow lines. The tape measure is located directly
above the cross-section to be measured and must not touch the water surface.
Identify the starting edge as either left edge of water (LEW) or right edge of water
(REW) when facing downstream. Determine the approximate width of the stream with
active stream flow, being sure not to include slack water areas. Hence, the edge of slack
water areas will be considered the edge of the stream.
Discharge measurements are taken at several verticals, defined as a point along the crosssection where water velocity is measured at a defined depth (or depths). Twelve to twenty
verticals will be targeted for streams <20 feet wide, whereas twenty to thirty verticals will
be targeted when stream width is >20 feet. To calculate the approximate spacing of
verticals, divide the stream width by the number of desired verticals. Importantly, the
stream discharge computed using the average velocity in one vertical shall not exceed
10% of the total stream discharge. Therefore, it may be necessary to space verticals more
closely together in areas that are deeper or that have a greater velocity than the majority
of the stream. Conversely, the spacing of verticals may be farther apart in areas that are
shallower or have lower velocity compared to the majority of the stream. Uniform
spacing across the tape measure will only be used if the stream is of relative uniform
depth and velocity regimes.
Although vertical spacing can vary, verticals will never be spaced less than 0.2 feet apart.
As a result of this minimum spacing, small streams with a flowing width of less than 2.2
feet will have less than 12 verticals and can have as few as one vertical during very low
stream flow.
A standard top-setting wading rod will be used to measure water depth and confirm the
proper location of the flow meter sensor within the water column. The wading rod will be
adjusted to the appropriate depth, which is marked in 0.1-foot increments along the rod. It
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is appropriate to further estimate depth to the 0.05-foot increment level, despite the
wading rod not being marked to this level. When water depth is ≤ 2.5 feet, velocity is
measured at 0.6 of the depth below the water surface at each vertical. A standard topsetting wading rod will automatically adjust the probe to this height. When water depth
is ≥ 2.5 feet, discharge is measured at 0.2 and 0.8 of the total depth below the water’s
surface at each vertical. To set the rod at the 0.2-depth, position the setting rod at half the
water depth. To set the rod at the 0.8-depth, position the setting rod at twice the water
depth. An average of these two readings will be used as the average velocity for the
vertical.
The wading rod will be held perpendicular to the water surface, and the instrument will
be parallel to the stream flow. The individual making the measurements will stand at least
1.5 feet away from the wading rod and 3 inches downstream of the tagline in a way that
alters the stream flow as little as possible. Rocks, logs, or other obstructions will not be
moved during the measurement process as this may cause the stream flow to change in an
area of the stream where velocity has already been measured. Once the process of
measuring velocity has begun, the stream will not be altered.
Record the location of the starting edge on the field data sheet (LEW or REW). If the
starting edge has a water depth, record this. No velocity measurements will be made at
the starting or ending edges. Facing upstream, place the wading rod behind the tape
measure at each vertical and record the location and stream depth. Velocity readings will
be averaged over a time period of 25s – 45s, depending on in-stream conditions. If the
water depth is ≥ 2.5 feet at a station, indicate the depth (0.2 or 0.8) associated with each
of the two velocity measurements. Record the ending edge (LEW or REW) as well as the
depth and velocity, if these exist.
If the stream cannot be safely waded or if a flow meter is not accessible, floats can be
used to estimate stream velocity needed for stream discharge computation. All
measurements using this procedure should be flagged as estimated on field data sheets
and on final data reports. The following steps are used in these conditions:
1. Find a long, relatively straight section of stream that allows a travel time of 20
seconds. A shorter time can be used if these conditions cannot be met.
2. Select two cross sections along the reach; one at the top and one at the bottom.
3. Measure the width of the stream at the cross sections and in a few areas
between the cross sections to obtain an average width. If the stream is not
wadeable, estimate the width. Record the width on the field data sheet.
4. Estimate how far an object will float in 20 seconds and stretch a tagline along
the stream bank to account for that distance. A distance of 30-50 feet is ideal.
A shorter run length may be used if these conditions cannot be met.
5. Based on the width, divide the stream into 2-3 longitudinal profiles. Measure
or estimate the depths at these profiles. On the field data sheet record the
nearest bank (REW/LEW) as 0. Record the farthest bank as the total width of
the stream.
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6. Have one person stand at the starting point on the tagline and a second person
stand at the point designated as the end of the run. The person at the end of the
run should use a stopwatch that can measure to tenths of a second.
7. The person at the starting point will throw a floating object (large stick,
orange, hedge apple, etc.) just upstream of the top cross section within the first
longitudinal profile area. When the object crosses the upstream cross section,
the person will say “start” and the person at the end of the run will start the
timer.
8. When the object crosses the downstream cross section, the person with the
timer will stop the timer.
9. Record the distance the object traveled and the number of seconds, to the
tenths of a second, the object took to travel that distance.
10. Repeat Steps 6-9 for the remaining profiles.
Velocity and flow area measurements must manually be recorded in a bound field book,
or on other appropriate field data sheets, using indelible, waterproof ink and waterproof
paper. Discharge values are to be calculated in the office according to the equations
specified in KDOW 2010b.
In addition to the discharge measurements collected, temporary water level gages
(pressure transducers with data loggers) will be installed at each of the six gaging
stations. These gages will be installed inside PVC pipe attached to a firmly anchored
stake or permanent instream structure with an elbow facing downstream to eliminate bias
from velocity-based pressures. If water levels become elevated enough to make streams
unwadeable and flow measurements unfeasible, the water level recorded by the loggers
may be useful in estimating stream discharge. The water level recorded by the loggers
could be used along with stream cross-section measurements and an estimation of surface
velocity (i.e. ‘float’ method) to estimate discharge during any un-wadeable parts of the
wet event.
The discharge data collected in this study will be used to improve modeling of the
watershed discharge based on the USGS gage and land use to incorporate the redirection
of the flow through the karst system. The results will be summarized in the “Karst
Hydrograph Characterization in the Wolf Run Watershed” final report due by May 31,
2012.
B2.2.2. Conductivity Survey
Trained volunteer samplers under the direction of the FOWR Sampling Coordinator will
conduct the conductivity survey. The Sampling Coordinator will schedule the survey
such that flow conditions at the USGS gage at the mouth of Wolf Run are between 0.5
and 5 cfs. In order to limit the temporal variations in the conductivity levels, efforts will
be made to conduct the entire survey within a one-week period unless interrupted by
precipitation. In the event of a precipitation event, the Sampling Coordinator will
reschedule the remaining sampling a minimum of 72 hours after the precipitation has
ceased.
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Each volunteer will be equipped with a GPS unit, digital camera, conductivity meter,
thermometer, and Conductivity Survey Field Datasheets (Appendix A). Volunteers will
each be assigned stream segments as shown in Table 9 and Exhibit 3. Before and after
monitoring activities, the volunteer shall calibrate the conductivity meters using a
standard of known value. The standard value, initial calibration reading (prior to
monitoring) and final calibration reading (after monitoring is completed) shall be
recorded on the datasheets along with the meter identification number (serial number).
Calibration and measurements with the conductivity meter shall be performed according
to the specifications in the instruction manual (see Appendix B).
Volunteers will begin the survey in the upstream portion of these reaches and work
downstream recording the time of measurement, latitude and longitude (NAD83 decimal
degrees), conductivity, temperature, and additional observations (including anomalous
conditions), if applicable, at each site. Photographs will be taken to document unusual
conditions with the photograph numbers indicated on the datasheet. Datasheets will be
used to document only one stream reach and monitoring day. If multiple datasheets are
necessary to cover a given stream segment, the order of these datasheets must be
indicated in the “Segment ID” by a dash and number value. For instance, if two
datasheets were used on Wolf Run segment 1A, these would be labeled 1A-1 and 1A-2 in
the “Segment ID” on the datasheet.
Datasheets and photos will be submitted to the FOWR Sampling Coordinator to be
compiled into a Microsoft Excel database. The FOWR Sampling Coordinator will
submit the electronic database, scanned copies of the field datasheets, and an electronic
photo library to Third Rock by November 30, 2011. Third Rock will use the data to
produce GIS images of the watershed, indicating hotspots. A final report “Conductivity
Survey of the Wolf Run Watershed” discussing the methods, results, and conclusions
based on the monitoring will be completed by December 31, 2011.
B2.2.3. Benthic Macroinvertebrate Collection
Sampling for benthic macroinvertebrates will be conducted according to the KDOW’s
Methods for Sampling Benthic Macroinvertebrate Communities in Wadeable Waters
(KDOW 2009c). All sites are headwater sites except W1, which is sampled for
macroinvertebrates under the MS4 permit and not under this project.
A collection event consists of a composited semi-quantitative sample and a composited
multi-habitat sample. Semi-quantitative samples will be collected from a known area in
order to indicate the macroinvertebrate community in the most productive habitat in the
stream niche (i.e., riffle). Multi-habitat samples are intended to identify other taxa
present in the stream that may not be collected in the semi-quantitative sampling. These
two sample types must be kept separate for effective diagnosis of impairment. A
summary of the collection techniques used for wadeable and headwater streams is shown
in Table 11 and further described in the following sections.
It is important to keep in-stream habitat intended for benthic macroinvertebrate sampling
intact and undisturbed until the single and multi-habitat samples have been collected.
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Therefore, field personnel must avoid walking through areas designated for collection of
benthic macroinvertebrates until sampling has been completed. Failure to use caution
could result in sample degradation.
After collections are completed, large sticks and leaves will be washed in the field,
inspected for organisms and discarded. Rocks will be elutriated and hand washed into a
bucket and 600μm sieve. This process will be repeated until a manageable amount of
debris and organisms (relative to size of sample container) can be preserved for
laboratory sorting. Samples may be partially field picked using a white pan and finetipped forceps. The sample container will be preserved with 95% ethanol. While at the
sampling location, all macroinvertebrate samples will receive a label. The label may be
placed in the sample jar (labels placed in the jar will be written in No. 2 pencil on
waterproof paper) and written directly on some portion of the jar. The label will include
the site number, if known, stream name, location, county, date sampled and the
collector’s initials.
TABLE 11 – SUMMARY OF SAMPLING METHODS FOR
MACROINVERTEBRATES
Technique
Sampling Device
1m2 kicknet / seine
Kicknet / seine
and wash bucket
Habitat
Semi-Quantitaive
Riffle
Replicates
Composited for
Wadeable Sites
Replicates
Composited for
Headwater Sites
4 x 0.25m2
4 x 0.25m2
3
N/A
3
3
3
3
3
N/A
N/A
N/A
3
3
3
3
3
N/A
15 total (5 each)
5 small boulders
3 to 6 linear meters
2 linear meters
Multi-Habitat Sweep
Undercut banks / roots
Sticks / Wood
Emergent vegetation
Bedrock / slabrock
J. americana beds
D-frame or
triangular dip net
and wash bucket
Leaf packs
Silt, sand, fine gravel
Aufwuchs sample
Rock pick
Wood sample
US #10 Sieve
300 μm nitrex
sampler / mesh
Fine-tipped
forceps and wash
bucket
All applicable
Riffle – Run –
Pool
Margins
Riffle – Run Pool
After sampling has been completed, all sampling gear will be thoroughly cleaned to
remove all benthic macroinvertebrates so that specimens are not carried to the next site.
The equipment shall be examined prior to sampling at the next site to ensure that no
benthic macroinvertebrates are present.
Habitat assessments will be performed at each of the macroinvertebrates sites. The
habitat assessment method is covered in Section B2.2.4.
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B2.2.3.1. Semi-Quantitative
In both headwater and wadeable streams, semi-quantitative sampling will consist of
taking four (4) 0.25m2 quadrat kick net samples from mid-riffle or the thalweg. This will
be accomplished using a 0.25 m2, 600μm mesh kick net, dislodging benthos by
vigorously disturbing the 0.25 m2 (20 x 20 in.) of substrate in front of the net. Large
rocks will be hand washed into the net. The contents of the net will then be washed, and
all four samples will be composited to yield a 1m2 semi-quantitative sample. The
composited sample will be partially field processed using a U.S. No. 30 sieve (600μm)
and wash bucket. Large stones, leaves and sticks will be individually rinsed and inspected
for organisms and then discarded. Small stones and sediment will be removed by
elutriation using the wash bucket and U.S. No. 30 sieve. This sample must be kept
separate from all other sub-habitat collections.
For headwater sites, two kick net samples will be allocated to each of two distinct riffles
(at minimum) that are separated by at least one pool or run. This will be done to help
reduce between-riffle variability. However, if there are several riffles located within the
reach, the sampling effort will be spread across the reach to give a comprehensive
evaluation of the entire community.
B2.2.3.2. Multi-Habitat
This method involves sampling a variety of non-riffle habitats with the aid of an 800 x
900μm mesh triangular or D-frame dip net. The habitats sampled and the number or size
of replicates will differ for headwater and wadeable sites, as shown in Table 11. Each of
these sub-habitat samples will be composited into one multi-habitat sample for each site.
Undercut Banks/Root Mats
These will be sampled by placing a large root wad into a triangular or D-frame dip net
and shaking vigorously. The contents will be removed from the dip net and placed into a
mesh wash bucket. If undercut banks are present in both run and pool areas, each will be
sampled separately with three (3) replicates for both headwater and wadeable streams.
Marginal Emergent Vegetation (exclusive of Justicia americana beds)
This habitat will be sampled by thrusting (i.e., “jabbing”) the dip net into the vegetation
for approximately 1m, and then sweeping through the area to collect dislodged
organisms. Material will then be rinsed in the wash bucket, and any sticks, leaves and
vegetation will be thoroughly washed and inspected before discarding. Three replicates
will be conducted. This sub-habitat must be sampled for wadeable sites and may be
sampled for headwater if present.
Bedrock or Slab-Rock Habitats
These habitats will be sampled by placing the edge of the dip net flush on the substrate,
and disturbing approximately 0.1m2 of area to dislodge attached organisms. Material will
be emptied into a wash bucket, rinsed, inspected for organisms, and discarded. Three
replicates will be conducted. This sub-habitat must be sampled for wadeable sites and
may be sampled for headwater if present.
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Justicia americana (water willow) Beds
These will be sampled by working the net through a 1m section in a jabbing motion. The
material will then be emptied into a wash bucket, and any J. americana stems will be
thoroughly washed, inspected and discarded. Three replicates will be conducted. This
sub-habitat must be sampled for wadeable sites and may be sampled for headwater if
present.
Leaf Packs
Leaf packs will preferably be collected from “conditioned” (i.e., not new-fall material)
material when possible. Samples will be taken from a diversity of habitats (i.e., riffles,
runs and pools) and placed into the wash bucket. The material will be thoroughly rinsed
to dislodge organisms, inspected and discarded. Three replicates will be conducted for
both headwater and wadeable sites.
Silt, Sand, and Fine Gravel
A U.S. No. 10 sieve will be used to sort larger invertebrates (e.g., mussels, burrowing
mayflies, dragonfly larvae) from silt, sand and fine gravel by scooping the substrate to an
approximate depth of 5cm. A variety of collection sites will be sampled in order to obtain
three (3) replicates in each substrate type where available (silt, sand and fine gravel).
This sub-habitat will be sampled for both headwater and wadeable sites.
Aufwuchs Sample
Small invertebrates associated with this habitat will be obtained by washing a small
amount of rocks, sticks, leaves, filamentous algae and moss into a medium-sized bucket
half filled with water. The material will then be elutriated and sieved with the nitrex
sampler/mesh. Three replicates will be conducted. This sub-habitat will be sampled only
for wadeable sites.
Rock Picking
Benthic macroinvertebrates will be picked from 15 rocks (large cobble/small boulders; 5
each from riffle, run and pool) in wadeable streams and 5 small boulders from pools in
headwater streams. Selected rocks will be washed in a bucket half filled with water and
then carefully inspected to remove organisms.
Wood Sample
For wadeable streams, pieces of submerged wood, ranging from roughly 3 to 6 meters
(10 to 20 linear feet) and ranging from 5–15 cm (2–6 inches) in diameter, will be
individually rinsed into the wash bucket. For headwater streams only 2 linear meters will
be sampled. Pieces of wood will be inspected for burrowers and crevice dwellers and
will be removed with fine-tipped forceps. Large diameter, well-aged logs will be
inspected and handpicked with fine-tipped forceps.
B2.2.3.3. Macroinvertebrate Identification and Analysis
Macroinvertebrate samples shall be delivered to Third Rock for identification according
to Laboratory Procedures for Macroinvertebrate Processing and Taxonomic Identification
and Reporting (KDOW. 2009b). After identification, macroinvertebrate sampling results
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will be evaluated through calculation of several community metrics prescribed by
KDOW 2008. Community metrics include taxa richness, EPT (mayfly, stonefly and
caddisfly) richness, total number of individuals, modified percent EPT individuals,
modified Hilsenhoff biotic index (mHBI), percent Ephemeroptera, percent primary
clingers, and percent Chironomidae plus Oligochaeta (aquatic worms). Results of
community metrics at each station will be combined to compute a Macroinvertebrate
Bioassessment Index (MBI) score, ranging from 0 (worst) to 100 (best). MBI scores will
be compared to scoring criteria developed by KDOW to arrive at water quality ratings of
Very Poor, Poor, Fair, Good, or Excellent. For wadeable streams (watersheds greater than
5 mi2) of the Bluegrass Bioregion, a MBI score below 20 is Very Poor, from 21 to 40 is
Poor, from 41 to 60 is Fair, from 61 to 79 is Good, and greater than 70 is Excellent. For
headwater streams (watersheds less than 5 mi2) of the Bluegrass Bioregion, a MBI score
below 18 is Very Poor, from 19 to 38 is Poor, from 39 to 50 is Fair, from 51 to 57 is Good,
and greater than 58 is Excellent (KDOW 2008).
Results from this project will be compared with Bluegrass Bioregion Criteria, reference
reach scores, and results from MS4 permit sampling at the mouth of Wolf Run. These
results and the results of the watershed habitat assessment monitoring will be combined
into a final report entitled “Assessment of Habitat and Macroinvertebrates in the Wolf
Run Watershed” due to the KDOW and LFUCG by December 31, 2011.
B2.2.4. Watershed Habitat Assessments
Habitat assessments will include a visual assessment of ten habitat parameters that
characterize the stream "micro scale" habitat, the "macro scale" features, and the riparian
and bank structure features that are most often influential in affecting the other
parameters. The method follows the US EPA’s Rapid Bioassessment Protocols for Use
in Wadeable Streams and Rivers (Barbour et al. 1999). Each of the parameters will be
evaluated on a “Condition Category” scale from 0 to 20. The categories within this scale
include “Optimal” for scores from 20 to 16, “Suboptimal” for scores from 15 to 11,
“Marginal” for scores from 10 to 6, and “Poor” for scores from 5 to 0. The score for each
parameter will be summed to produce a final habitat score (maximum 200).
For parameters 1 to 5, the habitat assessment will evaluate a composite of the entire
biological sampling reach. For parameters 6 to 10, an area beginning approximately 100m upstream of the sampling reach through the sampling reach will be evaluated as a
composite. The evaluator will face downstream when determining left and right bank.
For parameters 8 to 10, each bank will be scored independently from 10 to 0. At each
sampling site, results will be recorded on the High-Gradient Habitat Assessment Field
Data Sheet. Photographs will be taken to document upstream and downstream
conditions.
The following paragraphs summarize each of the ten parameters assessed.
Parameter #1 - Epifaunal Substrate/Available Cover
This metric measures the relative quantity and the variety of stable structures, such as
cobble, boulders, fallen trees, logs, branches, root mats, undercut banks, aquatic
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vegetation, etc., that provide refugia, feeding opportunities and sites for spawning and
nursery functions.
Optimal: >70% of substrate favorable for epifaunal colonization and fish cover;
mix of snags, submerged logs, undercut banks, cobble or other stable habitat
and at a stage to allow full colonization potential (i.e., logs/snags that are not
new fall and not transient).
Suboptimal: 40%-70% mix of stable habitat; well-suited for full colonization
potential; adequate habitat for maintenance of populations; presence of
additional substrate in the form of new fall, but not yet prepared for
colonization (may rate at the high end of the scale).
Marginal: 20%-40% mix of stable habitat; habitat availability less than desirable;
substrate frequently disturbed or removed.
Poor: <20% stable habitat; lack of habitat is obvious; substrate unstable or
lacking.
Parameter #2 Embeddedness
Embeddedness describes the extent to which rocks and snags are covered or sunken into
the silt, sand, mud or biofilms (algal, fungal or bacterial mats) of the stream bottom.
Generally, as rocks become embedded, the surface area available to macroinvertebrates
and fish (for shelter, spawning and egg incubation) is decreased; assess in the upstream or
central portions of riffles.
Optimal: Rocks are 0-25% surrounded by fine sediment. Layering of cobble
provides diversity of niche space.
Suboptimal: Rocks are 25%-50% surrounded by fine sediment.
Marginal: Rocks are 50%-75% surrounded by fine sediment.
Poor: Rocks are >75% surrounded by fine sediment.
Parameter #3 - Velocity/Depth Regime
The best streams in most high-gradient regions will have all of the following patterns of
velocity and depth: 1) slow-deep, 2) slow-shallow, 3) fast-deep and 4) fast-shallow; the
occurrence of these four patterns relates to the stream’s ability to provide and maintain a
stable aquatic environment. Investigators may have to scale deep and shallow depending
upon the stream size; a general guideline is 0.5 m between shallow and deep.
Optimal: All 4 regimes present.
Suboptimal: Only 3 of the 4 regimes present; if fast-shallow is missing, score
lower than if missing other regimes.
Marginal: Only 2 of the 4 regimes present; if fast-shallow or slow-shallow are
missing, score low.
Poor: Dominated by 1 regime (usually slow-deep).
Parameter #4 - Sediment Deposition
This metric measures the amount of sediment that has accumulated in pools and changes
that have occurred to the stream bottom as a result of deposition. This may cause the
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formation of islands, point bars (areas of increased deposition usually at the beginning of
a meander that increases in size as the channel is diverted toward the outer bank) or
shoals or result in the filling of runs and pools. Sediment is often found in areas that are
obstructed and areas where the stream flow decreases, such as bends. Deposition is a
symptom of an unstable and continually changing environment that becomes unsuitable
for many organisms. Examine bars/shoals and pool substrates within the biological
monitoring station, when assessing this parameter.
Optimal: Little or no enlargement of islands or point bars and less than 5%of the
bottom affected by sediment deposition.
Suboptimal: Some new increase in bar formation, mostly from gravel, sand or fine
sediment; 5%-30% of the bottom affected; slight deposition in pools.
Marginal: Moderate deposition of new gravel, sand or fine sediment on old and
new bars; 30%-50% of the bottom affected; moderate sediment deposits
apparent at most obstructions and slow areas, bends and pools.
Poor: Heavy deposits of fine material; increased bar development; more than
50% of the bottom changing frequently; pools almost absent due to substantial
sediment deposition.
Parameter #5 - Channel Flow Status
This metric measures the degree to which the channel is filled with water. The score will
change with the seasons. Estimate the percentage of the channel that is wet using the low
water mark.
Optimal: Water reaches base of both lower banks; minimal amount of channel
substrate exposed.
Suboptimal: Water fills >75% of the available channel or <25% of channel
substrate exposed.
Marginal: Water fills 25%-75% of the available channel; riffle substrates are
mostly exposed.
Poor: Very little water in channel; mostly present in pools.
Parameter #6 - Channel Alteration (Both Sheets)
This metric measures the large-scale, direct changes in the shape of the stream channel.
Channel alteration is present when 1) artificial embankments, rip-rap and other forms of
bank stabilization or structures are present, 2) the stream is very straight for significant
distances because of channelization, 3) dams and bridges are present that obstruct flow
and/or 4) dredging or other substrate mining activities are occurring or have occurred.
Optimal: Channelization or dredging absent or minimal; stream with normal
pattern.
Suboptimal: Some channelization present, usually in areas of bridge abutments;
evidence of past channelization (dredging, etc., >20 past years) may be
present, but recent channelization not present.
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Marginal: Channelization may be extensive; embankments or shoring structures
present on both banks; and 40%-80% of the stream reach channelized and
disrupted.
Poor: Banks shored with gabion or cement; >80% of the stream disrupted; in
stream habitat greatly altered or removed entirely.
Parameter #7 - Frequency of Riffles (or Bends)
This metric measures the sequence of riffles and thus the heterogeneity occurring in a
stream. Estimate riffle frequency by determining the ratio of distance between riffles
divided by the width of the stream. An average of the riffle ratios is determined for
biological monitoring stations and the upstream segment.
Optimal: Occurrence of riffles relatively frequent; ratio of distance between riffles
divided by the width of the stream <7:1 (generally 5 to 7); variety of habitat is
key; in streams where riffles are continuous, placement of boulders or other
large, natural obstruction is important.
Suboptimal: Occurrence of riffles infrequent; distance between riffles divided by
the width of the stream is between 7 and 15.
Marginal: Occasional riffle or bend; bottom contours provide some habitat;
distance between riffles divided by the width of the stream is between 15 and
25.
Poor: Generally all flat water or shallow riffles; poor habitat; distance between
riffles divided by the width of the stream is >25.
Parameter #8 - Bank Stability
This metric measures whether the stream banks are eroded or have the potential to erode.
Each bank is scored independently from 10-0.
Optimal: Banks stable; evidence of erosion or bank failure absent or minimal;
little potential for future problems; <5% of bank affected.
Suboptimal: Moderately stable; infrequent, small areas of erosion mostly healed
over; 5%-30% of the bank affected.
Marginal: Moderately unstable; 30%-60% of bank in reach has areas of erosion;
high erosion potential during floods.
Poor: Unstable; many raw, eroded areas; obvious bank sloughing; >60% of bank
has erosional scars.
Parameter #9 - Bank Vegetative Protection
This metric measures the amount of vegetative protection afforded to the stream and the
nearstream portion of the riparian zone. Each bank is scored independently from 10-0.
Determine what vegetative types (trees, understory shrubs, herbs and non-woody
macrophysics) are present on each bank. Those stream banks with different vegetative
types provide better erosion protection and provide more of a variety of allochthonous
food material. Native vegetation scores higher than invasive or non-native vegetation.
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Optimal: >90% of the stream bank surfaces and immediate riparian zones covered
by natural vegetation, including trees, understory shrubs, herbs and nonwoody macrophytes; vegetation disruption through grazing or mowing
minimal or not evident; almost all plants allowed to grow naturally.
Suboptimal: 70%-90% of the stream bank surfaces covered by native vegetation,
but one class of plants is not well-represented; disruption evident but not
affecting full plant growth potential to any great extent; more than one half of
the potential plant stubble height remaining.
Marginal: 50%-70% of the stream bank surfaces covered by vegetation;
disruption obvious; patches of bare soil or closely cropped vegetation
common; less than one half of the potential plant stubble height remaining.
Poor: <50% of the stream bank surfaces covered by vegetation; disruption is very
high; vegetation has been removed to 5 cm or less in average stubble height.
Parameter #10 - Riparian Vegetative Zone Width
This metric measures the width of the natural vegetation from the edge of the stream
bank through the riparian zone. The presence of old fields, paths, walkways, etc., in
otherwise undisturbed riparian zones may be judged to be inconsequential to highly
destructive to the riparian zone. Each bank is scored independently from 10-0. When
determining final scores, the age and density of the riparian vegetation should be
evaluated (e.g., A score of 9, instead of 10, should be given to a riparian zone that is over
20 m in width, but is dominated by 5-10 year old hardwood trees).
Optimal: Width of riparian zone >18 m; human activities (parking lots, roadbeds,
clear-cuts, lawns, pastures or crops) have not impacted the zone.
Suboptimal: Width of riparian zone 13-18 m; human activities have impacted the
zone only minimally.
Marginal: Width of riparian zone 6-12 m; human activities have impacted the
zone a great deal.
Poor: Width of riparian zone <6 m; little or no riparian zone due to human
activities.
All habitat assessments will be completed by October 31, 2011 and submitted to the
FOWR Sampling Coordinator. The FOWR Sampling Coordinator will compile these
results into an electronic Microsoft Excel database. The database, along with electronic
copies of the field datasheets will be submitted to Third Rock by November 30, 2011 for
incorporation into the final report discussing the results of the habitat assessments and the
macroinvertebrate survey.
B2.2.5. Hydrogeomorphic Assessment
Three types of measurements will be made in the hydrogeomorphic assessment: crosssections, longitudinal profiles, and pebble counts. These measurements will be made at
each of the nine hydrogeomorphic monitoring sites. Permanent monuments consisting of
rebar (0.75-inch rebar or similar material approximately 4 feet long) concreted within a
plastic pipe casing shall be installed at the permanent cross-section survey sites. A
monument shall be installed on both the right and left stream banks at least 10 ft back
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from the top of bank, indicating the extent of the measured cross-section and can serve as
surveying benchmarks. If installation of such monuments is not feasible, other
permanent monuments will be established. To facilitate profile relocation during the
second surveying period, the following actions shall be taken in the field:
1) monuments shall be marked with a piece of flagging or paint,
2) GPS points shall be recorded at monuments and any other locations that
would aid in site relocation,
3) photographs will be taken (for both relocation and to document the current site
conditions), and
4) notes will be recorded on site identification characteristics (e.g. bank
condition, distinguishing landmarks/features, and other pertinent data).
The methods for each of these sampling efforts are described below.
B2.2.5.1. Cross-Sections
Cross-sections to be surveyed will be located within riffle features and identified by
permanent monuments. Points will be taken frequently at horizontal stations within each
cross-section such that the surveying indicates all significant breaks in slope and provides
a thorough characterization of each cross-section (refer to USFS, 1994 for surveying
procedures). Equipment used will include a 50- or 100-ft surveying tape, laser level
(leveling accuracy < +/- 3mm/30m) on a tripod, and surveying rod. Data may be
recorded in RiverMorphTM software using a Rugged Reader Pocket PC or in a field
notebook. Surveying precision shall be +/- 0.01 ft for vertical readings and +/- 0.1 ft for
horizontal readings. Notes related to observed changes at various elevations within the
cross-section will be made. Each stream permanent cross-section will be surveyed twice,
once at the initial site visit following monument installation and approximately nine
months subsequent to first measurement. Differences between these two measurements
will allow estimation of channel change and if degradation is occurring, the erosion rate
can be calculated.
B2.2.5.2. Profiles
Representative stream longitudinal profiles will be taken over a distance that includes
approximately three riffle features at each of the nine hydrogeomorphic monitoring
stations. Permanent monuments on a designated bank and at least 10 feet back from the
top of bank will mark the upstream and downstream extents of the profiles and can serve
as benchmarks for surveying. Profile measurements will be taken within the stream
thalweg and will be of adequate frequency to identify all grade changes and facet slopes
within the profile (refer to USFS, 1994 for surveying procedures). Equipment used will
include a 100-ft surveying tape, laser level (leveling accuracy < +/- 3mm/30m) on a
tripod, and surveying rod. Data will be recorded in RiverMorphTM software using a
Rugged Reader Pocket PC or in a field notebook. Surveying precision shall be +/- 0.01 ft
for vertical readings and +/- 0.1 ft for horizontal readings. Locations of permanent crosssections and pebble count monitoring will be indicated within the recorded profile. Each
stream profile will be surveyed twice, once at the initial site visit following monument
installation and approximately nine months subsequent to first measurement. Differences
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between these two measurements will allow estimation of changes to channel bed
elevation, facet slope, and facet length.
B2.2.5.3. Pebble Counts
Reach-wide pebble counts will be collected within the stream where the longitudinal
profiles are taken at the nine hydrogeomorphic monitoring sites. If substrate does not
appear similar in all riffles, riffles with considerably coarser substrate that could be
indicative of a large rock fall will be avoided. Each reach pebble count will sample
within the riffles and pools proportional to the length of the reach comprised of riffles
and pools. Riffle and pool data will be kept separate, but can be combined later to
produce a reach average particle distribution. For the reach-wide pebble counts, particle
sampling will be completed along evenly spaced transects over the entire bankfull width
and consist of at least 100 particles (refer to Rosgen, 2008 and Bunte and Steven, 2001
for pebble count procedures). Since much of the bed material in these streams is
predominantly sand, silt, and clay or bedrock a higher number of particles sampled is not
indicated. If it is determined upon field investigation that a given hydrogeomorphic
monitoring site has a wide particle-size spectrum, at least 400 particles may be collected
for the reach-wide pebble count.
An active bed, riffle pebble count will also be collected within the permanent crosssection at each of the nine hydrogeomorphic monitoring sites. For the active riffle bed
count, particle sampling will be completed along evenly spaced transects over the active
bed width and consist of at least 100 particles (refer to Rosgen, 2008 and Bunte and
Steven, 2001 for pebble count procedures).
For all pebble counts, each transect will start on the same side of the stream and
collection will move from downstream to upstream. Sampling points will be spaced by at
least the Dmax particle size. The pebble count will end at the extent of a given transect,
not in an arbitrary location when a count of 100 particles is reached. If fine sediments
(sand/silt) are encountered and the thickness of the sediment layer is less than 0.5 inch,
then it will be appropriate to select the larger particle below the fines. Otherwise the
observation will be counted as fines (i.e. less than or equal to 2mm). Equipment used
will include a ruler (mm) or gravelometer (gravel template), with the gravelometer being
preferred. Data may be recorded in RiverMorphTM software using a Rugged Reader
Pocket PC or on a Pebble Count Datasheet (see Appendix A). Precision for pebble count
readings will be such that each data point measures within +/- 1 units of the narrative
particle description or +/- 0.5 phi units on the gravelometer. Each pebble count will be
performed twice, once at the initial site visit and approximately nine months subsequent
to first measurement. For each sampling event, particle size distributions and D50 values
will be computed and differences between these two measurements will allow estimation
of changes to channel substrate.
B2.2.6. Water Quality Monitoring
Water quality monitoring will be conducted at ten (10) monthly sampling events at a
minimum of twelve (12) sampling stations in the watershed during dry and wet
conditions. The sampling date within each month will be flexible such that at least two
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of the events will be considered ‘wet-weather’ and two of the events will be considered
‘dry-weather.’ The FOWR Sampling Coordinator shall work with the Third Rock
Sampling Coordinator to schedule sampling dates each month. A minimum notice of 48
hours is required prior to mobilization of the volunteer samplers.
During the monthly sampling, grab samples will be collected by volunteers and delivered
within six hours of collection to the LFUCG Town Branch laboratory for analysis of E.
coli, fecal coliform, total suspended solids, ammonia, nitrite, total dissolved solids,
alkalinity, and hardness. Samples for total phosphorus, ortho-phosphorus, total kjeldahl
nitrogen, and nitrate will be delivered to the Kentucky Geological Survey laboratory.
Volunteers will perform field measurements of turbidity, dissolved oxygen, specific
conductance, temperature, and pH. Third Rock will accompany the volunteers during
each event to conduct discharge monitoring and collect the field filtered orthophosphorus sample. Also due to the short time period in which wet-weather events can
be collected on the hydrographic rise, two wet-weather sampling events will be collected
solely by Third Rock staff. If for some reason, volunteers are not able to perform the
sampling, Third Rock will collect additional sampling events. Efforts will be made to
avoid collecting samples on Thursday, as this is the busiest day at the Town Branch
laboratory.
In addition to the monthly sampling, volunteers will collect an additional four events for
E. coli and fecal coliform to evaluate the geometric mean for the primary contact period.
A Third Rock staff member will accompany the volunteers during each event to conduct
discharge monitoring. Only flow and E. coli and fecal coliform will be collected during
these events. The LFUCG Town Branch laboratory will analyze the samples.
Subsequent to the completion of all sampling, results will be compiled and analyzed in a
final “Wolf Run Watershed Monitoring Report.” Further, this data will be used in
conjunction with other data collection efforts in the watershed to produce the loading
calculations for Chapter 4 of the Wolf Run Watershed Based Plan.
The methods to be utilized in performing these tasks are listed in the sections below.
B2.2.6.1. Flow Measurement
The procedure for flow measurement is explained in B2.2.1. Flow measurements will be
conducted according to the KDOW’s Measuring Stream Discharge Standard Operating
Procedure (KDOW 2010b).
Velocity and flow area measurements must manually be recorded in a bound field book,
or on other appropriate field data sheets, using indelible, waterproof ink and waterproof
paper. Discharge values will be calculated in the office according to the equations
specified in KDOW 2010b. One duplicate measurement will be recorded per sampling
event.
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B2.2.6.2. Grab Sampling
Grab sampling will be conducted according to the Kentucky Division of Water’s
Sampling the Surface Water Quality in Lotic Systems (KDOW 2011). The methods
specific to this collection effort are described below. One field duplicate sample will be
collected for all parameters per sampling event.
Samplers should put on powderless latex or nitrile gloves prior to sampling. Grab
samples should be collected in the centroid of flow in a section of stream in which
indicators of complete mixing are evident. The sampler should face upstream and
approach the site from downstream, ensuring that no disturbed streambed sediment
contaminates the sample. If additional work is planned upstream of the sample site, the
water samples must be taken first. Care should be taken not to displace the preservative
since sample bottles are pre-prepared.
If bottles are not pre-preserved, triple rinse the sample bottle with stream water prior to
sample collection.
When sampling, point the mouth of sample container
upstream/against the flow. Submerge the entire bottle and fill it with water. If the stream
is too shallow to fill the bottle while submerged, fill as much as possible while
submerged, ensuring the minimal amount for analysis is obtained. Also if the bottle
contains a preservative, angle the mouth so as not to spill the preservative while
collecting. Rinse the caps with sample water prior to capping the bottle. Samples should
be stored in containers that are free of potential contaminants. Sample bottles may be
placed inside sealed food-grade plastic bags prior to being stored on ice in coolers to
improve laboratory sorting and reduce potential cross-contamination.
For bacterial samples, fill bottles to the inscribed ‘fill line’. Do not pour off excess water.
If the bottle is filled above the 100ml fill line, the excess will be decanted during
laboratory analysis. If adequate volume is not obtained on first effort, do not reuse the
bottle. Use a new, un-contaminated bottle and repeat the procedure. Close and secure the
sample bottle lid immediately and preserve sample accordingly.
B2.2.6.3. Field Filtration
The collection of the ortho-phosphorus samples requires field filtration using a hand
pump. Third Rock will perform this field filtration within 15 minutes of sample
collection. In order to collect this field filtered sample, the stream sample will be
collected using the grab sample methodology. The funnel, funnel filter base and flask will
be triple rinsed with with DI water, and the hand pump, the inside of tubing and tweezers
will be single rinsed with DI water. Clean forceps will be used to place 0.45 μm paper
filter onto funnel filter base. The filter base will be attached to flask and the tubing from
the hand pump will be connected. 50 mL DI water will be poured into funnel, filtered,
rinsed and discarded. 50 mL of the stream sample water will be poured into funnel,
filtered, rinsed, and discarded. Then enough stream sample water will be poured into the
funnel to provide enough finished sample for rinsing the storage bottle and for analysis.
If the stream is particularly turbid, smaller amounts of the sample water will be used.
When 0.45 μm paper filter becomes excessively clogged, the filter will be removed with
forceps, discarded, and replaced with a fresh filter. Filtering will be continued until the
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required sample volume is achieved. The sample will be poured from the filter flask into
the sample bottle.
B2.2.6.4. In-situ Measurements
Volunteers will perform field measurements of turbidity, dissolved oxygen, specific
conductance, temperature, and pH. The sampling methods for temperature and pH are
specified in the Watershed Watch Water Chemistry Sampling Methods for Field
Chemistry and Lab Analysis (Appendix B). The temperature must be recorded to an
accuracy of 0.5 °C and pH to an accuracy of 0.5 SU. Instructions for the dissolved
oxygen and specific conductivity methods are specified in the instruction manuals
associated with the meter / test kits (see Appendix B). Turbidity will be evaluated
through a visual assessment and indicated as “clear”, “slightly turbid”, “turbid”, or
“other”. Results will be recorded on the chain-of-custody (Appendix A).
During the two wet-weather sampling events collected by Third Rock, Hydrolab
multimeters will be used to record the dissolved oxygen, specific conductance,
temperature, and pH. The procedures specified in In-situ Water Quality Measurements
and Meter Calibration Standard Operating Procedure (KDOW, 2009a) will be used in
these measurements. Results will be recorded in the field notebook.
At one site per sampling event, replicate measurements will be made on all in-situ
parameters except turbidity.
B3. Sample Handling and Custody Requirements
The sample handling and custody procedures are compatible with the SOP “Sample
Control and Management” (KDOW 2009c).
B3.1. Chain-of-Custody
Chain-of-Custody (COC) forms will be completed for all samples collected in the field
and will follow each sample throughout sample processing. A COC form is a controlled
document used to record sample information and ensure the traceability of sample
handling and possession is maintained from the time of collection through analysis and
final disposition. A sample is considered in custody if it is:
 In the individual’s physical possession
 In the individual’s sight
 Secured in a tamper-proof way by that individual, or secured in an area restricted
to authorized personnel
Example COCs that will be used in the collection are attached in Appendix A. All
information shall be documented on the COC in black or blue waterproof permanent ink
including field physical measurements and custody information.
The sampler shall initiate sample custody at the time the sample is collected. Field
custody documentation shall include:
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




Verification of sample identification
Number of sample bottles collected
Collection date
Collection time
Collector’s signature
The sampler shall maintain possession of the sample until custody is transferred to the
laboratory or another party. The COC shall accompany the sample from the time of
collection until it is relinquished. Field custody will be relinquished by signature, with
date and time, of the sample in the designated area on the COC.
B3.2. Sample Handling and Transport
The sampler will be responsible for sample handling in the field and transport of samples
to the laboratory. The sampler will collect the sample at the source following established
protocols. The sampler will be responsible for collecting the sample in appropriately
identified collection containers with the correct preservative, as applicable, and ensuring
that the container lid is secured tightly to prevent leakage or outside contamination.
Sample containers for chemical analysis shall be immediately placed in a cooler on ice to
maintain a temperature of 4±2° C for transport to the laboratory. Sample bottles shall be
placed in the cooler with lid side up in an organized manner per COC entry.
Sample coolers will be of adequate size to allow ice to surround all sample bottles. It is
the responsibility of the sampler to ensure that coolers are properly packed in the field
and that they have sufficient cooler space on their vehicle for their daily sample load.
Coolers shall be secured during transport such that significant disturbance of the samples
is avoided. Macroinvertebrate samples will be analyzed at Third Rock. The LFUCG
Town Branch laboratory and Kentucky Geological Survey laboratory will process the
chemical sampling parameters.
Upon receipt at the laboratory, the sample custodian shall review the COC for
completeness and accuracy. Anomalies shall be documented. The laboratory shall
measure sample temperature upon receipt, determine if sample aliquots have been placed
in appropriate bottles and properly preserved, and inspect the sample for proper
identification and bottle integrity; any discrepancies and/or bottle damage shall be
documented on the COC. If the hold time requirement is exceeded for any parameter, the
result is qualified and a re-sampling must be scheduled.
B3.3. Sample Labeling
Whenever possible during field preparation while in the office, sample bottles will be
labeled to prevent information omission. Bottles can be labeled in the field, as long as the
following minimum requirements for labeling are followed. All bottles must have the
following information recorded either on a sample tag or label affixed to the container, or
written directly on the container:
 Sample identification (unique site ID number)
 Date of collection
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





Time of collection (formatted in military time, or indicate am or pm)
Type of analysis requested
Type of sample (grab, composite, semi-quantitative, multi-habitat)
Media (surface water, biological-macroinvertebrates)
Preservative (ice, acidification, etc.)
Collector’s initials
For macroinvertebrate samples, the stream name and location will also be documented on
the label.
B3.4. Sample Designation
Sampling technicians will be responsible for recording the unique sample identification,
as well as the date and time of the collection on each sample bottle. The unique sampling
event code will follow the following format:
SAMPLE ID = W##-YYMMDD
Where:
W## is Unique Site Identifier (0-12 or “DD” for Duplicate)
YYMMDD is the date in year (YY), month (MM), day (DD) format.
As indicated above, duplicate samples will be indicated as such in the site identifier of
“WDD.” The time of collection will not be indicated on the chain-of-custody so that the
laboratory is blind as to the sampling location it corresponds with. This information shall
be recorded in the field notebook during collection so that results can be compared after
analysis.
B4. Analytical Methods Requirements
USEPA methodology must be used for all analysis as applicable. Detection limits for all
parameters must be at a sensitivity level to compare to Kentucky water quality standards.
The requirements for all methods and detection limits are specified in Table 6. SOPs for
the chemical laboratory methods are specified in Price 2009.
Third Rock’s macroinvertebrate identification laboratory will follow laboratory protocols
for benthic macroinvertebrate sample processing, identification and data reporting per
KDOW (2009b, 2008) with the following exceptions:
 All samples will be logged into Third Rock's Macroinvertebrate Laboratory
Information Management System (MacLIMS) upon receipt.
 Sample identification date will be maintained in MacLIMS.
 Taxonomic QA/QC dates (if applicable) will be noted on individual QA/QC
forms and maintained electronically in the Project File.
 Initials of the applicable party completing each task associated with sorting,
identification, or quality control will be noted electronically in MacLIMS or on
associated QA/QC forms.
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 QA checks will be documented on applicable forms and maintained in associated
project files. These forms include the Macroinvertebrate Sample Sorting
Efficiency Form, Macroinvertebrate Sample Taxonomy Precision Form, and
Macroinvertebrate Sample Taxonomic and Enumeration Efficiency Form.
B5. Quality Control Requirements
B5.1. Field Quality Controls
Field quality control checks for water chemistry will be collected at a frequency of one
duplicate every sampling event. Field duplicates must be randomly determined from the
12 sites and recorded on field datasheets or project logbooks. The field controls shall be
performed as follows.
A random number table (see Appendix A) will be used to select one sampling site such
that all sample parameters are duplicated. Two separate samples will be collected for
each parameter. The samples will be collected at the same time and at the same location.
One sample will be labeled as usual, and the other sample will have the site name
indicated as a “duplicate.” On a form separate from the COC, the site from which the
duplicates were collected will be documented. In addition to the duplicate grab sample
collection, duplicate in-situ measurements and flow measurements will also be made at
this site.
Duplicate habitat assessments will be performed by all volunteer samplers as well as by
Third Rock biologists at the six macroinvertebrate sampling sites. These duplicate
measurements will be used to evaluate precision amongst volunteers conducting habitat
assessments.
During the karst characterization, a minimum of five percent of measurements will be
duplicated.
B5.2. Macroinvertebrate Taxonomic Quality Controls
Ten percent of all sorting pans will be checked by a second sorter to assure that samples
have been picked thoroughly. These samples will be selected randomly using the
MacLIMS database programming. This check is documented on the Taxonomic &
Enumeration Efficiency Form.
Five percent of all identified samples will be re-identified to insure QA/QC by a second
taxonomist. These samples will be selected randomly using the MacLIMS database
programming. Ninety percent or greater composition comparability (e.g., abundance and
richness) is the target success criteria. If there is less than 90 percent comparability
between the taxonomists, then taxonomy must be reconciled by both taxonomists and a
third taxonomist, if deemed necessary. This quality control process shall be documented
on the Macroinvertebrate Sample Taxonomy Precision Form and Macroinvertebrate
Sample Taxonomic and Enumeration Efficiency Form and included in the monitoring
report.
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B5.3. Chemical Laboratory Quality Controls
Laboratory quality controls will be analyzed as specified in the SOPs listed in Quality
Assurance Plan (QAP) and Standard Operating Procedures (SOPs) (Price 2009). These
controls include method blanks, matrix spikes, calibration check samples, laboratory
replicates, and other method specified controls. The frequencies of analysis for these
standards are all specified by the individual methods.
B6. Instrument / Equipment Testing, Inspection, Calibration, and Maintenance
Requirements
Laboratory instrumentation will be maintained according to the SOPs listed in Table 5.
Field sampling equipment will be maintained as specified in KDOW 2009b and Table 12.
The record of inspection, calibration, and maintenance will be recorded in a instrument
logbook maintained by the sampler. For sampling nets and bottles, inspection will ensure
that the items are free from contamination, in good condition, and adequate for use.
TABLE 12 – FIELD EQUIPMENT CALIBRATION AND MAINTENANCE
Equipment
Name/Type
Equipment
Purpose
Inspection
Frequency
Type of
Inspection
Multimeter for
Temperature, pH,
Dissolved Oxygen
and Conductivity
Physicochemical
Monitoring
Before each
collection
event
Overall
condition/
battery power
EC Conductivity
PockeTesters
Conductivity
Measurement
Hand pumps, filter
funnel, tubing, and
flasks
Field Filtration
Flow Meter
Discharge
Monitoring
Before each
collection
event
Before each
collection
event
Before each
collection
event
Overall
condition/
battery power
3 solution
wash, overall
condition
Overall
condition/
battery power
Cleanliness,
program
settings
Overall
condition
Good
condition
Macroinvertebrate
Sampling Nets
Karst
Characterization
Stage Measurement
Macroinvertebrate
Sampling
Sample Bottles
Sample Collection
TROLL®
Dataloggers
Before use
Before each
use
Before
collection
Standard or
Calibration
Instrument
Used
Person
Responsible
Calibration
standards, user
manual
Sampler
Calibration
standards, user
manual
Sampler
N/A
N/A
Sampler
Annual
Manufacturer
Sampler
N/A
N/A
Sampler
N/A
N/A
Sampler
N/A
N/A
Sampler
Calibration
Frequency
Before each
field
sampling
event
Before and
after each
uses
Volunteer samplers will calibrate the conductivity meters before and after each sampling
event. Two standards will be used to calibrate the meters with results recorded on the
chain-of-custody or Conductivity Survey Field Datasheet (Appendix A).
Third Rock calibrates flow meters annually through the manufacturer. The multimeters
to be used will be calibrated according to manufacturers instructions prior to each
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sampling event using a three point pH calibration and a one point conductivity
calibration. All results will be recorded in the instrument logbook.
The filter funnel, tubing, and flask used in field filtration will be cleaned prior to
sampling or weekly at maximum. To clean the equipment, three clear HDPE washbasins
will be used. One washbasin will be labeled “Detergent Wash”, one “Acid Solution” and
the final “DIW.” The detergent used for cleaning equipment must be certified phosphatefree. All washbasins used during the cleaning process must be pre-cleaned following the
same procedures:
1. Detergent Wash and Tap Water Rinse
a. Put on powderless nitrile gloves
b. Place equipment in basin labeled “Detergent Wash” and soak equipment in a
tap water/detergent mix for 30 minutes
c. Fill tubing with solution and keep submerged for 30 minutes
d. Scrub exterior and interior surfaces of equipment
e. Rinse thoroughly with warm tap water to remove detergent residue
2. Acid Soak and Rinse
a. Put on a new pair of gloves
b. Place equipment and tubing into a washbasin labeled “Acid Solution”; for
pieces of equipment that contain metal parts, skip to Step 3.
c. Fill washbasin with 5% HCl solution (ACS trace-element grade HCL; 5% by
volume in DIW).
d. Soak for 30 minutes; Stir solution occasionally to promote the detachment of
organic and inorganic contamination from the equipment
3. DI Water Rinse
a. Put on a new pair of gloves
b. Place equipment and tubing into a washbasin labeled “DIW”
c. Rinse all equipment and tubing with DI water
d. Place onto a clean surface to dry
4. Clean Equipment Storage
a. Place clean equipment in plastic storage bags
b. Double bag tubing
B7. Data Management
In order to ensure that project objectives are achieved, data must be collected and
managed in a manner that will protect and ensure its integrity. The data collected under
this project will be produced under standardized procedures and forms, where
practicable.
All field data will be recorded using black or blue indelible ink. Mistakes on field data
sheets will be crossed out with one line (so the information is still discernible), with the
initials and date of the person making the correction. The correct information will then
be recorded legibly on another line, or above or below the original info. If a separate
sheet is necessary for new information, the original sheet will be attached to the new
sheet, and initialed and dated.
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Data collected by volunteers shall be submitted to the Ken Cooke, FOWR Sampling
Coordinator. He shall review the data for any nonconformances, and enter the data into
electronic databases and forward the electronic databases and electronic copies of the
original datasheets to the QA Manager. He will be responsible for storing all original
copies of the volunteer field data collected.
Chemical laboratory analytical results and internal laboratory logbook documentation
will be the responsibility of David Price, Laboratory Director. Macroinvertebrate
laboratory results and metric calculations will be the responsibility of Bert Remley,
Macroinvertebrate Laboratory Chief Taxonomist. Upon completion of laboratory
analysis, results shall be forwarded electronically to Steve Evans, QA Manager.
Data collected by Third Rock samplers and staff will be maintained by Steve Evans, QA
Manager. He will receive the laboratory reports and review the data for conformance to
the requirements of this QAPP and will subsequently send the results to the KDOW and
LFUCG. He will be responsible for sending all hardcopy and electronic copies of data
reports to the LFUCG and KDOW, as well as maintaining previously submitted data. All
final reports will receive a technical peer review and a grammatical / formatting review
prior to submission.
All raw data, documentation, and records shall be retained. Correspondence and other
documentation relating to interpretation and evaluation of data collected, analyzed, or
processed shall also be retained. The retention period is a minimum of three years
subsequent to grant completion. All data maintained on Third Rock computers will be
supported by a daily backup and archival system. Hard copy files will be stored onsite
under secure conditions.
No data shall be publicly disseminated unless first reviewed and approved for release by
the Project Team including the Grantee Project Manager, QA Manager, and FOWR
Sampling Coordinator. Prior to that time, data will be managed by Third Rock.
Subsequent to final approval, Project Team members may distribute the results as
appropriate.
SECTION C – ASSESSMENT AND OVERSIGHT
Assessment and response actions are necessary to ensure that this QAPP will be
implemented as approved. For a general summary of these assessments see Table 13. If
at any time a project team member finds an error or non-conformance in the QAPP, the
QAPP will be revised and redistributed to those on the distribution list subsequent to
approval. The KDOW QA officer may freely review all field and laboratory techniques
as requested. Any identified problems will be corrected based on recommendations by
the KDOW QA Officer.
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TABLE 13 – WATERSHED ASSESSMENT AND MANAGEMENT REPORTS
Type
Frequency
Purpose
QAPP Revision
As necessary
Address non-conformances
or errors in the QAPP
KDOW Audit
As requested
Ensure conformance to
project objectives
Laboratory
Demonstration
of Performance
Prior to initial
analysis
Laboratory
Internal Audits
Parties Responsible For
Performing
Responding
Reporting
Method
Project Team
Members
QA Manager
Distribution of
amended QAPP
KDOW
Parties of concern
Corrective
Action Response
Ensure analyst is capable of
performing the method to
specifications.
Laboratory QA
Director
Laboratory
Analysts
Internal Lab
documentation
Annually, at
minimum
Ensure conformance to
methods, regulations, and
procedures.
Laboratory QA
Director
Laboratory
Analysts
Internal Lab
documentation
Progress
Assessment
Quarterly
Evaluate the status on
project related objectives
and concerns
Grantee Project
Manager, or
designee
KDOW
Section 319(h)
Nonpoint Source
Project Progress
Report
Volunteer Field
Sampling Audit
Once during
project
Assess volunteer sampler
conformance to proper
sampling and
documentation protocols.
QA Manager, or
designee
Analytical
Results Review
Subsequent to
each sampling
event
Quality
Evaluation
Once, End of
Project
Evaluate the conformance
of laboratory data to project
DQOs
Evaluate the quality
assurance and compare the
data produced to project
DQIs
Volunteer
Field Sampling
Samplers, Trainers Audit Checklist
Laboratory Data
Quality
Checklist
QA Manager, or
designee
Laboratory QA
Director
QA Manager, or
designee
KDOW QA
Final Monitoring
Officer, Grantee
Reports
Project Manager
To ensure conformance with this QAPP and the applicable regulations, certifications, and
methods by which the laboratories operate, the laboratories will perform several
assessment measures. To ensure that analysts are capable of performing the requested
analytical methods to specifications, each analyst must acceptably demonstrate this
ability prior to conducting sample analyses. The analyst must conduct four replicate
analyses of a known standard and achieve precision and accuracy equal to or better than
the acceptance ranges for laboratory duplicates and laboratory control samples,
respectively. The laboratory QA Director or his appointee on an annual basis will
perform internal audits. The findings of the audits, both positive and negative, will be
documented, and the corrective response to the cited deviations will be made. Corrective
actions will be submitted to the auditing body for review and approval.
The QA Manager is responsible for the overall conformance of field personnel to the
procedures, protocols, and methods established by this QAPP and internal project related
procedures. The QA Manager will conduct at least one audit of the volunteer samplers
field activities and documentation including calibration and maintenance of field
equipment and sample collection techniques. Deviations found in such assessments will
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be reported to the samplers and documented using the Field Sampling Audit Checklist
(Appendix A).
Upon receipt of the results, a review of the laboratory data shall be performed by the QA
Manager or his designee to ensure that the project DQOs have been satisfied. The
Laboratory Data Quality Checklist (see Appendix A) shall be utilized to document this
review. The final reports for each of the monitoring activities will include an evaluation
of the quality assurance and will compare the data produced under the water quality
monitoring to the data quality indicators listed herein.
Quarterly Section 319(h) Nonpoint Source Project Progress Report will be submitted to
the KDOW by the Grantee Project Manager to document the progress on the project
milestones.
SECTION D – DATA VALIDATION AND USABILITY
D1. Data Review, Validation and Verification
Data review is the internal examination to check if data has been recorded, transmitted,
and processed correctly. Data verification is the process of evaluating whether the data
meets method, procedural, or contractual specifications. Data validation is the review of
the quality of the data based on the specific DQIs indicated in this QAPP.
The sampler will perform data review for all field data initially before submitting to the
laboratory. Upon submission to the laboratory, the laboratory will review the COC for
completeness and document any non-conformances on the COC.
For the chemical laboratory data, the laboratory analyst will initially conduct the review,
and the data will be peer reviewed by another analyst or capable reviewer. Data will be
reviewed according to the laboratory QA Manual and the method specific SOP for data
entry, calculations, and transformations as well review of quality control criteria. If
deviations are noted, corrective actions will be taken with verification of both the
reviewer and the original data collector. If consensus cannot be reached, the data will be
rejected. During verification and validation of the data, all data that does not meet the
DQIs listed in this QAPP will be qualified or rejected. A list of the type of qualifiers that
may be applied to this data is listed in Table 14. All qualified data will be evaluated
according to the actions listed.
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TABLE 14 – DATA QUALIFIERS AND RESPONSE
Definition
Analyte detected in
associated Method Blank
Diluted out
Holding time exceeded
Estimated value
Matrix Spike and/or Matrix
Spike Duplicate Recovery
outside acceptance limits
Laboratory Control Sample
outside acceptance limits
Sample received exceeding
proper temperature or
preservation criteria
The analyte was analyzed for
but not detected
Analyte exceeded calibration
range
Replicate / Duplicate
precision outside of
acceptance limits
Calibration criteria exceeded
Action To Be Taken
Reject results. Indicates all, or a portion of, the amount found in a
sample may be due to laboratory sources.
Accept results. Indicates a dilution to overcome matrix effects caused
other analytes of interest to be diluted out of range. Normal
quantitation is not available.
Reject results. Method required holding time is exceeded.
Accept results when used to indicate result is below the project
reporting limit, but above the Method Detection Limit (MDL).
Acceptable results if associated Laboratory Control Sample is
acceptable (No qualifier). Indicates matrix is adversely affecting the
extraction or digestion of the analyte. If the Matrix Spike recovery is
below acceptable limits, it may be likely that the reported results for
the associated samples may be underestimated. Conversely, if the
Matrix Spike results are high, it may be likely that the reported
results for the associated samples may be overestimated.
Reject results. Indicates that the laboratory system is out of control.
Reject results. Indicates preservatives or temperature requirements
have not been met and the bias on the sample result is unknown.
Accept results. Indicates that the result is less than the reporting
limit
Accept results. Only reported in instances in which the calibration
curve is exceeded and the sample cannot be reanalyzed.
Reject results, unless it occurs on a matrix spike duplicate. Indicates
the precision is outside of normal acceptance criteria due to a lack of
homogeneity or other factor.
Reject results. Indicates that the laboratory system is out of control.
D2. Validation and Verification Methods
The QA Manager will use the Field Sampling Audit Checklist and the Laboratory Data
Checklist (Appendix A) to document the conformance of the data to this QAPP. This
review will be submitted to the KDOW along with the data. The QA Manager will be
responsible for making any final decisions concerning data quality and acceptability.
All final reports will receive an internal peer review to evaluate the content, calculations,
and data analysis in the report. The reports will also undergo an internal grammatical
review to look for grammatical errors and formatting. Lastly, the final report will receive
a review from the Project Team prior to submission to the KDOW to ensure that all
project objectives are achieved.
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D3. Reconciliation with User Requirements and Data Quality Objectives
In the final report, descriptions of all relevant background information, summary, water
body details, monitoring results, recommended solutions, and implementation plans will
be detailed. Included in this document will be an overall assessment of the data quality
and the uncertainty involved in the results.
SECTION E. - REFERENCES AND CITATIONS
Barbour, M.T., J. Gerritsen, B.D. Snyder, and J.B. Stribling. 1999. Rapid Bioassessment
Protocols for Use in Streams and Wadeable Rivers: Periphyton, Benthic
Macroinvertebrates and Fish. Second Edition. EPA 841-B-99-002. USEPA,
Office of Water, Washington, D.C.
Bunte, Kristin; Abt, Steven R. 2001. Sampling surface and subsurface particle-size
distributions in wadable gravel-and cobble-bed streams for analyses in sediment
transport, hydraulics, and streambed monitoring. Gen. Tech. Rep. RMRS-GTR74. Fort Collins,CO: U.S. Department of Agriculture, Forest Service, Rocky
Mountain Research Station. 428 p.
Compton, M.C., G.J. Pond, and J.F. Brumley. 2003. Development and application of the
Kentucky Index of Biotic Integrity (KIBI). Kentucky Department for
Environmental Protection, Division of Water, Frankfort, Kentucky.
Harrelson, C.C., C.L. Rawlins, and J.P. Potyondy. 1994. Stream channel reference sites:
An illustrated guide to field technique. General Technical Report RM-245. Fort
Collins, CO: U.S. Department of Agriculture, Forest Service, Rocky Mountain
Forest and Range Experiment Station. 61p.
In-Situ Inc. 2006. Level TROLL® Operator’s Manual. www.in-situ.com
Kentucky Division of Water (KDOW). 2004. Project Final Report Guidelines for Clean
Water Act §319(h)-Funded Projects. Kentucky Energy and Environment Cabinet
Department for Environmental Protection Division of Water Watershed
Management Branch Nonpoint Source Section.
KDOW. 2008. Methods for Assessing Biological Integrity of Surface Waters in Kentucky.
Kentucky Department of Environmental Protection.
KDOW. 2009a. In-situ Water Quality Measurements and Meter Calibration Standard
Operating Procedure. Kentucky Department for Environmental Protection,
Division of Water, Frankfort, Kentucky. DOWSOP03014
KDOW. 2009b. Laboratory Procedures for Macroinvertebrate Processing and
Taxonomic Identification and Reporting. Kentucky Department of Environmental
Protection.
________________________________________________________________________
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Date: April 11, 2011
KDOW. 2009c. Methods for Sampling Benthic Macroinvertebrate Communities in
Wadeable Waters. Kentucky Department for Environmental Protection, Division
of Water, Frankfort, Kentucky.
KDOW. 2010a. 2010 Integrated Report to Congress on the Condition of Water
Resources in Kentucky: Volume II. 303(d) List of Surface Waters. Kentucky
Energy and Environment Cabinet Division. Frankfort, Kentucky.
KDOW. 2010b. Measuring Stream Discharge Standard Operating Procedure. Kentucky
Department for Environmental Protection, Division of Water, Frankfort,
Kentucky. DOWSOP03019
KDOW. 2011. Sampling the Surface Water Quality in Lotic Systems. Kentucky
Department for Environmental Protection, Division of Water, Frankfort,
Kentucky. DOWSOP03015
KDOW. Watershed Watch Water Chemistry Sampling Methods for Field Chemistry and
Lab Analysis. http://www.lrww.org/training/chem-test.pdf
Kentucky Waterways Alliance and the Kentucky Division of Water. 2010. Watershed
Planning Guidebook for Kentucky Communities. 1st ed. Kentucky Waterways
Alliance and the Kentucky Division of Water.
LaMotte. Dissolved Oxygen Water Quality Test Kit Instruction Manual. Code 7414 /
5860. LaMotte Company. Chestertown, Maryland. www.lamotte.com
Lexington-Fayette Urban County Government (LFUCG). 2008. Stormwater Quality
Management Program (SWQMP) for Lexington-Fayette Urban County
Government. Tetra Tech, Inc, LFUCG Department of Public Works and
Department of Environmental Quality.
Oakton Instruments. Waterproof TDSTestr and ECTestr Series Instructions.
http://www.4oakton.com/Manuals/ConductivityTDS/TDS_ECTestrmnl.pdf
Price, David J. 2009. Quality Assurance Plan (QAP) and Standard Operating Procedures
(SOPs). Lexington-Fayette Urban County Government Division of Water Quality
Town Branch Laboratory.
Recker, S.A. and Meiman, J., 1990, Remedial investigations status report, Shop-N-Go
Food Mart: Delta Environmental Consultants, Lexington, KY, unpublished report,
18 p.
Rosgen, D.L. 2008. River Stability Field Guide. Wildland Hydrology, Pagosa Springs,
CO.
________________________________________________________________________
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Spangler, L.E., 1992, Unpublished groundwater tracing data: University of Kentucky,
Department of Geology.
United States and Commonwealth of Kentucky v. Lexington-Fayette Urban county
Government, March 14, 2006, Consent Decree, Lodged in the United States
District Court, Eastern District of Kentucky, Central Division at Lexington,
Related to Civil Action No. 5:06-cv-00386. Accessed November 2010 at
http://lexingtonky.gov/modules/ShowDocument.aspx?documentid=3572
United States Environmental Protection Agency (US EPA). March 2008, Handbook for
Developing Watershed Plans to Restore and Protect Our Waters, Office of Water,
Nonpoint Source Control Branch, EPA 841-B-08-002
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APPENDIX A – DATASHEETS
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Volunteer Monitoring Participant Agreement
Please fill this out, keep one and turn one in.
Please Print Name:
Best Phone
Organization:
Shipping Address:
City State Zip
E-mail:
I, to the best of my ability, will: (Check those that apply)
1. ___Conduct Field Chemistry. (D.O, pH, Conductivity, Temp)
2. ___Collect Grab Samples,
3. ___Take Photographs
4. ___Read and follow the Project Quality Assurance Project Plan
5. ___Return equipment when I "retire" from the project.
6. Equipment checked out to me: KIT#__________
10.
ID#
Lab Analysis Sample Sights Requested
Stream Name
Sampling Site (Map Reference)
11.____I understand and agree that the stream can be a dangerous place,
the project organizers
cannot protect me from slick banks, high water, snakes, falling timber and other hazards. I will ask
permission before crossing private property. I will be on my guard and exercise due caution when
handling chemicals. The information above is correct to the best of my knowledge.
SIGNED____________________________________DATE________________
Lead Instructor Verification: This is to verify that the volunteer listed above has completed
the modules I have initialed below:
Instructor Name(print)____________________________________Date________________
Initial
Module:
Grab Sample Collection
Field Chemistry
Initial
Module:
Project Logistics and Quality Assurance
Habitat Assessment
Note to lead instructor: Return a copy of this form to your project registrar or to the Water Watch Office at 400 Fair Oaks
Plaza Frankfort KY 40601 Att: JoAnn Palmer 1-800-928-0045 Ext 473
HABITAT ASSESSMENT FIELD DATA SHEET — HIGH GRADIENT STREAMS (FRONT)
STREAM NAME:
LOCATION:
INTERMITTENT
STREAM WDTH (FT):
DEPTH (FT):
PERENNIAL
STATION #:
RIVERMILE:
COUNTY:
LAT:
LONG:
RIVER BASIN:
CLIENT:
EPHEMERAL
STATE:
PROJECT NO.
INVESTIGATORS/CREW:
FORM COMPLETED BY:
DATE:
REASON FOR SURVEY:
TIME:
Parameters to be evaluated in sampling reach
Habitat
Parameter
Condition Category
Suboptimal
Marginal
Optimal
Poor
1. Epifaunal
Substrate/
Available Cover
Greater than 70% of
substrate favorable for
epifaunal colonization and
fish cover; mix of snags,
submerged logs, undercut
banks, cobble or other
stable habitat and at stage
to allow full colonization
potential (i.e., logs/snags
that are not new fall and not
transient.
40-70% mix of stable
habitat; well suited for full
colonization potential;
adequate habitat for
maintenance of populations;
presence of additional
substrate in the form of
newfall, but not yet
prepared for colonization
(may rate at high end of
scale).
20-40% mix of stable
habitat; habitat availability
less than desirable;
substrate frequently
disturbed or removed.
Less than 20% stable
habitat; lack of habitat is
obvious; substrate unstable
or lacking.
SCORE:
20
15
10
5
2. Embeddedness
Gravel, cobble, and boulder
particles are 0-25%
surrounded by fine
sediment. Layering of
cobble provides diversity of
niche space.
Gravel, cobble, and boulder
particles are 25-50%
surrounded by fine
sediment.
Gravel, cobble, and
boulder particles are 5075% surrounded by fine
sediment.
SCORE:
20
15
10
3. Velocity/Depth
Regime
All four velocity/depth
regimes present (slowdeep, slow-shallow, fastdeep, fast-shallow). (Slow
is < 0.3 m/s, deep is > 0.5
m.)
Only 3 of the 4 regimes
present (if fast-shallow is
missing, score lower than if
missing other regimes).
Only 2 of the 4 habitat
regimes present (if fastshallow or slow-shallow
are missing, score low).
SCORE:
20
15
10
4. Sediment
Deposition
Little or no enlargement of
islands or point bars and
less than 5% of the bottom
affected by sediment
deposition.
Some new increase in bar
formation, mostly from
gravel, sand or fine
sediment; 5-30% of the
bottom affected; slight
deposition in pools.
Moderate deposition of
new gravel, sand or fine
sediment on old and new
bars; 30-50% of the
bottom affected; sediment
deposits at obstructions,
constrictions, and bends;
moderate deposition of
pools prevalent.
Heavy deposits of fine
material, increased bar
development; more than
50% of the bottom changing
frequently; pools almost
absent due to substantial
sediment deposition.
SCORE:
20
15
10
5
5. Channel Flow
Status
Water reaches base of both
lower banks, and minimal
amount of channel
substrate is exposed.
Water fills > 75% of the
available channel; or <25%
of channel substrate is
exposed.
Water fills 25-75% of the
available channel, and/or
riffle substrates are mostly
exposed.
Very little water in channel
and mostly present as
standing pools.
20
15
10
5
SCORE:
19
19
19
19
19
18
18
18
18
18
17
17
17
17
17
16
16
16
16
16
14
14
14
14
14
13
13
13
13
13
12
12
12
12
12
11
11
11
11
11
J:\Forms and Templates\Technical Data Sheets and Checklists\Streams\Habitat Assessment Field Data Sheet - high.doc
9
9
9
9
9
8
8
8
8
8
7
7
7
7
7
6
4
3
2
1
0
Gravel, cobble, and boulder
particles are more than 75%
surrounded by fine
sediment.
6
5
4
3
2
1
0
1
0
Dominated by 1
velocity/depth regime
(usually slow-deep).
6
6
6
5
4
4
4
3
3
3
2
2
2
1
1
0
0
01/31/08
HABITAT ASSESSMENT FIELD DATA SHEET — HIGH GRADIENT STREAMS (BACK)
Parameters to be evaluated in sampling reach
Habitat
Parameter
Condition Category
Suboptimal
Marginal
Optimal
Poor
6. Channel
Alteration
Channelization or dredging
absent or minimal; stream
with normal pattern.
Some channelization
present, usually in areas of
bridge abutments; evidence
of past channelization, i.e.,
dredging, (greater than past
20 yr) may be present, but
recent channelization is not
present.
Channelization may be
extensive; embankments
or shoring structures
present on both banks;
and 40 to 80% of stream
reach channelized and
disrupted.
SCORE:
20
15
10
7. Frequency of
Riffles (or bends)
Occurrence of riffles
relatively frequent; ratio of
distance between riffles
divided by width of the
stream < 7:1 (generally 5 to
7); variety of habitat is key.
In streams where riffles are
continuous, placement of
boulders or other large,
natural obstruction is
important.
Occurrence of riffles
infrequent; distance
between riffles divided by
the width of the stream is
between 7 to 15.
SCORE:
20
15
8. Bank Stability
(score each bank)
Banks stable; evidence of
erosion or bank failure
absent or minimal; little
potential for future
problems. < 5% of bank
affected.
Note: determine left
or right side by facing
downstream.
19
19
18
18
17
17
16
16
14
14
13
13
12
11
12
11
Moderately stable;
infrequent, small areas of
erosion mostly healed over.
5-30% of bank in reach has
areas of erosion.
9
8
7
Banks shored with gabion
or cement; over 80% of
the stream reach
channelized and
disrupted. Instream
habitat greatly altered or
removed entirely.
6
5
4
3
2
1
0
Occasional riffle or bend;
bottom contours provide
some habitat; distance
between riffles divided by
the width of the stream is
between 15 to 25.
Generally all flat water or
shallow riffles; poor
habitat; distance between
riffles divided by the width
of the stream is a ration of
> 25.
10
5
9
8
7
6
Moderately unstable; 3060% of bank in reach has
areas of erosion; high
erosion potential during
floods.
4
3
2
1
0
Unstable; many eroded
areas; "raw" areas
frequent along straight
sections and bends;
obvious bank sloughing;
60-100% of bank has
erosional scars.
SCORE: (LB)
Left Bank
10
9
8
7
6
5
4
3
2
1
0
SCORE: (RB)
Right Bank
10
9
8
7
6
5
4
3
2
1
0
9. Vegetative
Protection (score
each bank)
More than 90% of the
streambank surfaces and
immediate riparian zone
covered by native
vegetation, including trees,
understory shrubs, or nonwoody macrophytes;
vegetative disruption
through grazing or mowing
minimal or not evident;
almost all plants allowed to
grow naturally.
SCORE: (LB)
Left Bank
10
9
8
7
6
5
4
3
2
1
0
SCORE: (RB)
Right Bank
10
9
8
7
6
5
4
3
2
1
0
10. Riparian
Vegetative Zone
Width (score each
bank riparian zone)
Width of riparian zone >18
meters; human activities
(i.e., parking lots, roadbeds,
clear-cuts, lawns, or crops)
have not impacted zone.
SCORE: (LB)
Left Bank
10
9
8
7
6
5
4
3
2
1
0
SCORE: (RB)
Right Bank
10
9
8
7
6
5
4
3
2
1
0
70-90% of the streambank
surfaces covered by native
vegetation, but one class of
plants is not wellrepresented; disruption
evident but not affecting full
plant growth potential to any
great extent; more than
one-half of the potential
plant stubble height
remaining.
Width of riparian zone 1218 meters; human activities
have impacted zone only
minimally.
50-70% of the streambank
surfaces covered by
vegetation; disruption
obvious; patches of bare
soil or closely cropped
vegetation common; less
than one-half of the
potential plant stubble
height remaining.
Width of riparian zone 612 meters; human
activities have impacted
zone a great deal.
Less than 50% of the
streambank surfaces
covered by vegetation;
disruption of streambank
vegetation is very high;
vegetation has been
removed to 5 centimeters
or less in average stubble
height.
Width of riparian zone <6
meters: little or no riparian
vegetation due to human
activities.
TOTAL SCORE:
J:\Forms and Templates\Technical Data Sheets and Checklists\Streams\Habitat Assessment Field Data Sheet - high.doc
01/31/08
Conductivity Survey Field Datasheet
Genral Info:
Stream Name
Date
Segment ID
Sampler Name
Measurements to be made at least 100
ft apart in each segment starting
upstream and working downstream.
Cond Meter ID
Calibration:
Known Value
Known Value
Time
(Indicate AM or PM)
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
Additional Notes:
714 µS/cm Initial Calibration
1438 µS/cm Initial Calibration
Latitude
Longitude
N DD.DDDDDD W DD.DDDDDD
Final Calibration
µS/cm Final Calibration
Conductivity
Temperature
(µS/cm)
(°C)
µS/cm
µS/cm
µS/cm
Additional Comments
Pebble Count Datasheet
Date
Survey Crew
Stream
Station
Particle
Description
Size (mm)
SILT/CLAY
silt/clay
<0.062
SAND
very fine
0.062 - 0.125
fine
0.125 - 0.25
med
0.25 - 0.5
coarse
0.5 - 1
very coarse
1-2
very fine
2-4
0.16 - 0.22
fine
4 - 5.7
0.22 - 0.31
fine
5.7 - 8
0.31 - 0.44
med
8 - 11.3
0 44 - 0
0.44
0.63
63
med
11 3 - 16
11.3
0.63 - 0.89
coarse
16 - 22.6
0.89 - 1.26
coarse
22.6 - 32
1.26 - 1.77
very coarse
32 - 45
1.77 - 2.5
very coarse
45 -64
small
64 - 90
3.5 - 5.0
small
90 - 128
5.0 - 7.1
large
128 - 180
7.1 - 10.1
large
180 - 256
small
256 - 362
14.3 - 20
small
362 - 512
20 - 40
med
512 - 1024
40 - 80
large
1024 - 2048
Size (in)
0.04 - 0.08
0.08 - 0.16
2.5 - 3.5
10.1 - 14.3
GRAVEL
COBBLE
BOULDER
BEDROCK
Total
bedrock
Riffle
Particle Count
Pool
Other
Total
%
Cum %
CHAIN OF CUSTODY
Analytical Report to:
LFUCG
Division of Water Quality
Town Branch Laboratory
301 Lisle Industrial Avenue, Lexington,
Kentucky, 40511
(859) 425-2416
* Matrix Code
* * Preservation Type
SW - Surface Water
I, ST, SA I, ST I, ST
** Preservative Code
4oz
P
4oz
P
32oz
P
Requested Lab Analysis
On-Site/Field Measurements
Sample Location
Final
Final
Matrix *
µS/cm
µS/cm
Collection
Date
Collection
Time
Grab /
Comp
Filt
d
Filt'd
Y/N
Alkalinity, TSS,
TDS, Hardness,
Nitrite
µS/cm
µS/cm
# of Containers Per
Analysis
W01-11 __ __ __ __
Wolf Run @ Old Frankfort Pike
SW
G
N
1
1
1
1
W02-11 __ __ __ __
McConnel Branch @ Prestons Cave
SW
G
N
1
1
1
1
W03-11 __ __ __ __
Wolf Run @ Valley Park
SW
G
N
1
1
1
1
W04-11 __ __ __ __
Vaughn's Branch @ Valley Park
SW
G
N
1
1
1
1
W05-11 __ __ __ __
Cardinal Run @ Devonport Dr
SW
G
N
1
1
1
1
W06-11 __ __ __ __
Wolf Run @ Wolf Run Park
SW
G
N
1
1
1
1
W07-11 __ __ __ __
Vaughn's Branch @ Pine Meadow Park
SW
G
N
1
1
1
1
W08-11 __ __ __ __
Vaughn's Branch @ Picadome
SW
G
N
1
1
1
1
W09-11 __ __ __ __
Wolf Run @ Faircrest Dr
SW
G
N
1
1
1
1
W10-11 __ __ __ __
Springs Branch @ Faircrest Drive
SW
G
N
1
1
1
1
W11-11 __ __ __ __
Big Elm Tributary @ Harrodsburg Rd
SW
G
N
1
1
1
1
W12-11 __ __ __ __
Wolf Run @ Lafayette Pkwy
SW
WDD-11 __ __ __ __
Duplicate
SW
Relinquished By:
Date / Time
----
----
Received By:
G
N
1
1
1
1
G
N
1
1
1
1
Date / Time
Turb
bidity (Visual)
Sample I.D.
Initial Calibration
Initial Calibration
Ammonia
Known: 714 µS/cm
Known: 1438 µS/cm
E. coli
Conductivity Calibration:
Fecal coliform
Nitrite to RL of 0.03 mg/L
o
Ammonia to RL of 0.05mg/L
8oz
P
Tem
mperature ( C)
SA - H2SO4
ST - Na2S2O3
I - Ice
Comments:
I
Container Size/Type
Spe
ecific Conductance
(umh
ho/cm)
Turnaround Time Required: 30 Working Days
pH (S.U.)
Methodology Required: 40CFR Part 136
[email protected]
Marcia L. Wooton
Third Rock Consultants, LLC
2526 Regency Road
Suite 180
Lexington, KY 40503
859-977-2000
Field Remarks:
Diss
solved Oxygen (mg/L)
Client: Third Rock Consultants, LLC
Project Name: Wolf Run Watershed Based Plan
Project # : KY10-030
Project Contact (sampling): Marcia L. Wooton
Phone # : 859-977-2000
Collected By:
Page 1 of 1
1=cle
ear, 2=slightly turbid,
3=turrbid, or indicate if “other”
COC#
See Field Notes for Duplicate ID
Temp. Upon Receipt (C): __________ Measured By:________
Containers Properly Preserved: (Yes / No)
Headspace: (Yes / No / NA)
Bottles Intact: (Yes / No)
COC Seals Intact: (Yes / No / NA)
Additional Documentation Attached: (Yes / No)
Original COC To Laboratory (Accompany Samples & Report)
COC Copy - TRC Project File
COC Copy - TRC Laboratory Services Coordinator
sje 04/11/11
COC#
CHAIN OF CUSTODY
SW - Surface Water
Turnaround Time Required: 30 Working Days
I, FF, SA
** Preservative Code
FF - Field Filter
HA - HCl
NA - HNO3
SA - H2SO4
SH - NaOH
ST - Na2S2O3
I - Ice
Collection
Date
Collection
Time
8oz
P
Filt'd
Y/N
G
N
1
Sample Location
Matrix *
W01-11 __ __ __ __
Wolf Run @ Old Frankfort Pike
SW
W02-11 __ __ __ __
McConnel Branch @ Prestons Cave
SW
G
N
W03-11 __ __ __ __
Wolf Run @ Valley Park
SW
G
N
W04-11 __ __ __ __
Vaughn's Branch @ Valley Park
SW
G
W05-11 __ __ __ __
Cardinal Run @ Devonport Dr
SW
W06-11 __ __ __ __
Wolf Run @ Wolf Run Park
SW
W07-11 __ __ __ __
Vaughn's Branch @ Pine Meadow Park
SW
W08-11 __ __ __ __
Vaughn's Branch @ Picadome
SW
W09-11 __ __ __ __
Wolf Run @ Faircrest Dr
SW
G
W10-11 __ __ __ __
Springs Branch @ Faircrest Drive
SW
W11-11 __ __ __ __
Big Elm Tributary @ Harrodsburg Rd
SW
W12-11 __ __ __ __
Wolf Run @ Lafayette Pkwy
SW
Duplicate
SW
Relinquished By:
Date / Time
----
----
Received By:
8oz
P
32oz
P
# of Containers Per
Analysis
Grab /
p
Comp
Sample I.D.
ID
WDD-11 __ __ __ __
I, SA
Requested Lab Analysis
Orthophosphorus
Comments:
Ortho-Phosphorus to RL of 0.05 mg/L
Phosphorus to RL of 0.02 mg/L
Nitrate to RL of 0.03 mg/L
TKN to RL of 0.5 mg/L
I
Container Size/Type
Total
phosphorus,
TKN
Methodology Required: 40CFR Part 136
Analytical Report to:
[email protected]
Marcia L. Wooton
Third Rock Consultants, LLC
2526 Regency Road
228 Mining & Mineral Resources Building, University of Kentucky
Suite 180
Lexington, Kentucky, 40506-0107
Lexington, KY 40503
859-977-2000
(859) 323-0555
* Matrix Code
* * Preservation Type Field Remarks:
Nitrate
Client: Third Rock Consultants, LLC
Project Name: Wolf Run Watershed Based Plan
Project # : KY10-030
Project Contact (sampling): Marcia L. Wooton
Phone # : 859-977-2000
Collected By:
Page 1 of 1
1
1
1
1
1
1
1
1
N
1
1
1
G
N
1
1
1
G
N
1
1
1
G
N
1
1
1
G
N
1
1
1
N
1
1
1
G
N
1
1
1
G
N
1
1
1
G
N
1
1
1
G
N
1
1
1
Date / Time
Comments
See Field Notes for Duplicate ID
Temp. Upon Receipt (C): ___ Measured By:____________
Containers Properly Preserved: (Yes / No)
Headspace: (Yes / No / NA)
Bottles Intact: (Yes / No)
COC Seals Intact: (Yes / No / NA)
Additional Documentation Attached: (Yes / No)
Original COC To Laboratory (Accompany Samples & Report)
COC Copy - TRC Project File
COC Copy - TRC Laboratory Services Coordinator
sje 04/11/11
Macroinvertebrate Sample Chain of Custody
Project Information Sheet
Project Administrator:
Client Name:
Project Number:
Sampling Site Location/Stream Name:
System Type:
Due Date:
County:
EcoRegion:
State:
Total Number of Samples:________ Total Number of Containers:
Reporting Requirements: __ Laboratory Data Sheet; __ Excel Spreadsheet; __ MBI Calculations
via : __ e-Submittal; __ Hardcopy; __ Both
Samples Relinquished By: ___________________ Date/Time: ____________ Sample Received By: ____________________ Date/Time: ____________
Samples Relinquished By: ___________________ Date/Time: ____________ Sample Received By: ____________________ Date/Time: ____________
Comments/Special Instructions: _______________________________________________________________________________________________
Sample Reference ID
Qualitative or
Quantitative
Latitude
Longitude
Collected
By
Collection
Date
Sample
Type
Field
Preservative
# of
Containers
Per Sample
- Continue on Reverse for More Samples System Type: Headwater Stream; Wadeable Stream; Large River; Lotic; Other _________
EcoRegion: Bluegrass; Mountain; Pennyroyal; Mississippi Valley-Interior River Lowlands; Other _________
Sample Type: KN KickNet; TK Traveling Kick; MH Multihabitat; S Surber; HD Hester-Dendy Multiplate; HDD HD Deep; HDS HD Shallow; OT Other_______; NA Not Available
MacLIMS: Client Setup/Login By _______________ Date __________; Reported By _______________ Date __________; Invoiced By _______________ Date __________
Sample Reference ID
Qualitative or
Quantitative
Latitude
Longitude
Collected
By
Collection
Date
Sample
Type
Field
Preservative
# of
Containers
Per Sample
Third Rock Consultants, LLC
Macroinvertebrate Sample Sorting Efficiency Form
Client Name: ___________________________
SampleID: _ _____________________________
Third Rock Project #: ___________________
Original Sorter:
Date Sorted:
# of Grids Sorted:
# of Organisms Originally Sorted:
# organisms
originally sorted
÷
(
Resorted By:
Date Resorted:
# of Grids Sorted:
# Additional Organisms Recovered:
# additional
organisms recovered
# organisms
originally sorted
+
)
=
% Sorting Efficiency
Additional Organisms Located
Taxon
Number
Total:
Comments:
Reviewed By: _________________________________ Date: _____________
Updated 4/29/10
Third Rock Consultants, LLC
Macroinvertebrate Sample Taxonomic & Enumeration Efficiency Form
Original Taxonomist:
Original Date Completed:
# Organisms Enumerated (Taxonomist 1):
Client Name: ___________________________
Sample ID: ___________________________
Third Rock Project #: ________________
Second Taxonomist:
Review Date Completed:
# Organisms Enumerated (Taxonomist 2):
Percent Difference in Enumeration (PDE) =
(n1 – n2) ÷ (n1 + n2) x 100 = % Difference in Enumeration (PDE)
n1 = # organisms counted by Taxonomist 1
n2 = # organisms counted by Taxonomist 2
Percent Taxonomic Disagreement (PTD) =
PTD = [ 1 – (comppos ÷ N)] X 100
Comppos = number of taxonomic agreements (see Taxonomic Comparison Form)
N = total number of organisms
Comments:
Reviewed By: __________________________________ Date: ___________
Updated 4/29/10
Third Rock Consultants, LLC
Macroinvertebrate Sample Taxonomy Precision Form
Client Name: ____________________
Sample ID: ______________________
Third Rock Project #: ______________
Taxon
Taxonomist 1
Totals:
Taxonomist 2
0
Reviewed By: _______________________________ Date: __________
# Agreements
0
0
Updated 4/29/10
Field Sampling Audit Checklist
General Information:
Project No. / Description:________________________________________
Field Personnel:________________________
Date:__________________
Reviewer: ________________________
Methods Reviewed:
Associated SOP, manual, or quality standard:
Evaluation:
For each DQI, indicate whether the field personnel met the project objectives. Document in the comments columns the
actual specification for that DQI and explain any non-conformances.
Data Quality Indicator (DQI)
Conforms?*
Y
N
Comments
Precision
Agreement among repeated
measurements of the same property
under identical, or substantially similar
conditions, random error. (i.e.
duplicates)
Bias
Systematic error or persistent distortion
of a measurement in one direction
Accuracy
Overall agreement of a measurement to
a known value; includes a combination of
precision and bias. (i.e. difference from
known)
Representativeness
Qualitative term expressing the degree to
which a portion accurately and precisely
represents the whole (i.e. where
sampled, conditions)
Completeness
Amount of valid data needed to be
obtained (Ensure all data is recorded and
all datasheets are filled out in their
entirety)
Sensitivity
Capability to discriminate between
measurement responses representing
different levels of variable interest (i.e.
reporting limit, lowest unit of measure)
Interference
Elimination of distorting or inhibiting
effects on the measurement
Training
Documented evidence of meeting all
necessary training requirements
*Yes / No – Indicates that the task performed in conformance / out of conformance with the project’s DQI. If the DQI is
not relevant to the task, write N/A in the comments column.
Page 1 of 2
Field Sampling Audit Checklist
Water Quality Sample Collection:
Data Quality Indicator (DQI)
Conforms?*
Y
Comments
N
Samples correctly labeled?
Storage and preservation?
Equipment Used:
Equipment Name
Last Calibration
By / Date
Conforms?
Comments
Other Comments:
*Yes / No – Indicates that the task performed in conformance / out of conformance with the project’s DQI. If the DQI is
not relevant to the task, write N/A in the comments column.
Page 2 of 2
Laboratory Data Quality Checklist
General Information:
Project No. / Description:________________________________________
Laboratory: ________________________
Date:__________________
Reviewer: ________________________
Data Reviewed:
Evaluation:
For each DQI, indicate whether the laboratory data met the project objectives. If not, document non-conformances in the comments column
and the action to be taken by data users. Attach additional sheets if necessary for documentation.
Data Quality Indicator (DQI)
Conforms?*
Y
N
Comments
Precision
Agreement among repeated
measurements of the same property
under identical, or substantially similar
conditions, random error. (i.e. duplicates,
splits)
Accuracy
Overall agreement of a measurement to
a known value; includes a combination of
precision and bias. (i.e. difference from
known laboratory control standard of
matrix spike)
Sensitivity
Capability to discriminate between
measurement responses representing
different levels of variable interest (i.e.
reporting limit, method blank
contamination)
Completeness
Amount of valid data required (Ensure all
data is reported with a useable result)
Preservation/Handing
Analyzed within hold time and properly
preserved
Interference
Elimination of distorting or inhibiting
effects on the measurement
Other
Describe non-conformance
*Yes / No – Indicates that the is in conformance / out of conformance with the project’s DQI. If the DQI is not relevant
to the data, write N/A in the comments column.
Page 1 of 1
APPENDIX B – STANDARD OPERATING PROCEDURES
________________________________________________________________________
Wolf Run Watershed
Page 67 of 67
Revision No.:0
Based Plan
Date: April 11, 2011
Lexington-Fayette Urban County Government
Division of Water Quality
Town Branch Laboratory
Quality Assurance Plan (QAP)
and
Standard Operating Procedures (SOPs)
July 8, 2008
Revised September 16, 2009
David J. Price, Ph.D.
Laboratory Supervisor
TB Laboratory QAP
Revision 1
September 16, 2008
Table of Contents
FORWARD..................................................................................................................................... 1
Section 1.......................................................................................................................................... 2
Quality Policy Statement ............................................................................................................ 2
1.1
Quality Policy Statement ............................................................................................ 2
1.2
Specific Objectives ..................................................................................................... 2
1.3
QAP Availability ........................................................................................................ 2
Section 2.......................................................................................................................................... 3
Laboratory Organization and Staff Responsibilities................................................................... 3
2.1
Laboratory Organization............................................................................................. 3
2.2
Staff Responsibilities .................................................................................................. 3
2.3
Current Personnel........................................................................................................ 3
Section 3.......................................................................................................................................... 4
Organizational Charts ................................................................................................................. 4
3.1
Laboratory's Place in Company and Laboratory Organization................................... 4
3.2
Laboratory’s Organization .......................................................................................... 4
Section 4.......................................................................................................................................... 5
4.1
Scope........................................................................................................................... 5
4.2
QAP Documentation Control Procedures................................................................... 5
4.3
SOP Documentation Control Procedures.................................................................... 5
4.4
Forms/Data Sheets Documentation Control Procedures............................................. 5
Section 5.......................................................................................................................................... 6
Laboratory's Approved Signatories ............................................................................................ 6
5.1
Approved Signatories.................................................................................................. 6
Section 6.......................................................................................................................................... 7
General Quality Control Procedures and Practices..................................................................... 7
6.1
Documentation of Test Methods and Laboratory Practices........................................ 7
6.2
General Quality Control Guidelines ........................................................................... 7
6.3
Method Blanks ............................................................................................................ 7
6.4
Matrix Spikes .............................................................................................................. 8
6.5
Laboratory Control Samples ....................................................................................... 8
6.6
Matrix Spike Duplicates and Sample Duplicates........................................................ 9
6.7
Quality Control Charts and Tabulations ..................................................................... 9
6.8
Initial Demonstration of Method Performance Studies (IDMP)............................... 10
6.9
Method Detection Limits (MDL).............................................................................. 11
6.10 Internal Audits .......................................................................................................... 11
6.11 Analytical Quality Control Standards....................................................................... 12
6.12 Reagents.................................................................................................................... 12
6.13 Glassware.................................................................................................................. 12
6.14 Laboratory Pure Water.............................................................................................. 12
Section 7........................................................................................................................................ 13
Verification Practices................................................................................................................ 13
7.1
Performance Evaluation (PE) Testing ...................................................................... 13
7.2
Standard Reference Materials ................................................................................... 13
7.3
Internal Quality Control Programs............................................................................ 13
a
TB Laboratory QAP
Revision 1
September 16, 2008
Section 8........................................................................................................................................ 15
Equipment Procedures for Calibration, Verifications, and Maintenance ................................. 15
8.1
Equipment Calibration .............................................................................................. 15
8.2
Balances .................................................................................................................... 16
8.3
pH Meters.................................................................................................................. 16
8.4
Conductivity Meters.................................................................................................. 16
8.5
Thermometers ........................................................................................................... 16
8.6
Refrigerators ............................................................................................................. 16
8.7
Incubators.................................................................................................................. 17
8.8
Ovens ........................................................................................................................ 17
8.9
Other Laboratory Equipment .................................................................................... 18
Section 9........................................................................................................................................ 19
Test Methods and SOPs ............................................................................................................ 19
9.1
Scope......................................................................................................................... 19
9.2
Standard Operating Procedures (SOPs) .................................................................... 19
Section 10...................................................................................................................................... 20
The Laboratory’s Physical Facilities Including Services and Resources ................................. 20
Section 11...................................................................................................................................... 22
Procedures for Reviewing New Work ...................................................................................... 22
11.1 Scope......................................................................................................................... 22
11.2 Unknown Samples .................................................................................................... 22
11.3 Contract Analyses ..................................................................................................... 22
Section 12...................................................................................................................................... 23
Sample Acceptance and Receipt Policy.................................................................................... 23
12.1 Scope......................................................................................................................... 23
12.2 Sample Acceptance Requirements............................................................................ 23
12.3 Verification of Preservation...................................................................................... 25
12.4 Actions for Deviations from the Lab's Sample Acceptance Requirements .............. 25
12.5 Laboratory Log of Received Samples....................................................................... 26
Section 13...................................................................................................................................... 27
Sample Tracking and Storage Procedures ................................................................................ 27
13.1 Scope......................................................................................................................... 27
13.2 Sample Tracking Record........................................................................................... 27
13.3 Proper Storage and Avoidance of Cross-contamination of Samples ........................ 27
13.4 Security of Samples .................................................................................................. 27
13.5 Sample Disposal........................................................................................................ 27
Section 14...................................................................................................................................... 28
Record Keeping, Data Review and Reporting Procedures ....................................................... 28
14.1 Scope......................................................................................................................... 28
14.2 Records Maintained by the Laboratory..................................................................... 28
14.3 Records of Standards and Analytical Reagents ........................................................ 29
14.4 Computerized Data Storage ...................................................................................... 30
14.5 Administrative Records ............................................................................................ 30
14.6 Laboratory Record Entries and Change of Entries ................................................... 30
14.7 Record Retention ...................................................................................................... 31
14.8 Raw Data Associated with Sample Analysis ............................................................ 31
b
TB Laboratory QAP
Revision 1
September 16, 2008
14.9 Test Data Reports...................................................................................................... 31
14.10
Electronic Data Transfer ....................................................................................... 32
14.11
Sample Disposal Records ..................................................................................... 32
14.12
Waste Disposal...................................................................................................... 32
Section 15...................................................................................................................................... 33
Corrective Action Policies and Procedures............................................................................... 33
15.1 Scope......................................................................................................................... 33
15.2 Identification of Discrepancies ................................................................................. 33
15.3 Staff Responsible for Investigation and Corrective Action ...................................... 33
15.4 Documentation and Review of Corrective Action.................................................... 34
Section 16...................................................................................................................................... 35
Procedures for Permitting Departures from Documented Policies and Procedures ................. 35
16.1 Scope......................................................................................................................... 35
16.2 Requests for and Approval of Departures................................................................. 35
16.3 Use of Analytical Methods Not Approved by KYDEP or U.S. EPA ....................... 35
16.4 Reporting of Non-compliant Data ............................................................................ 35
Section 17...................................................................................................................................... 36
Procedures for Dealing with Complaints.................................................................................. 36
17.1 Scope......................................................................................................................... 36
17.2 Complaint Receipt Procedures.................................................................................. 36
17.3 Complaint Investigation Procedures ......................................................................... 36
17.4 Complaint Response Procedures............................................................................... 36
Section 18...................................................................................................................................... 37
Procedures for Protecting Confidentiality and Proprietary Rights ........................................... 37
18.1 Procedure for Protecting Confidentiality .................................................................. 37
18.2 Examples of Confidential Data................................................................................. 37
18.3 Procedure for Open-Records Requests ..................................................................... 37
Section 19...................................................................................................................................... 38
Procedures for Internal Audits .................................................................................................. 38
19.1 Annual Internal Audit ............................................................................................... 38
19.2 Corrective Actions Regarding the Annual Internal Audit ........................................ 38
19.3 Notification of Invalid Data ...................................................................................... 38
Section 20...................................................................................................................................... 39
20.1 Scope......................................................................................................................... 39
20.2 Correcting Deficiencies in the QAP ......................................................................... 39
Section 21...................................................................................................................................... 40
Training and Personnel Requirements ...................................................................................... 40
21.1 Scope and Training Objectives ................................................................................. 40
21.2 Training Required for Laboratory Supervisor .......................................................... 40
21.3 Training Required for Quality Assurance Officer .................................................... 40
21.4 Training Required for Technicians ........................................................................... 40
21.5 Additional Training for Technicians......................................................................... 41
21.6 Substituting Experience or Education to Meet Training Requirements ................... 41
21.7 Instrument Training Requirements ........................................................................... 41
21.7 Training Records....................................................................................................... 41
Section 22...................................................................................................................................... 42
c
TB Laboratory QAP
Revision 1
September 16, 2008
Glossary .................................................................................................................................... 42
Section 23...................................................................................................................................... 51
Bibliography ............................................................................................................................. 51
APPENDIX A LABORATORY ANALYSIS LIST ............................................................... A-1
Table A1. Town Branch Laboratory Analysis List................................................................. A-2
Table A2. West Hickman WWTP Analysis List .................................................................... A-5
Table A3. Blue Sky WWTP Analysis List ............................................................................. A-7
Table A4. Data Provided to the Laboratory by Operations Staff ........................................... A-8
APPENDIX B STAFF RESPONSIBILITIES ........................................................................ B-1
APPENDIX C CURRENT PERSONNEL AND LAB APPROVED SIGNATURES ........... C-1
TOWN BRANCH LABORATORY STANDARD OPERATING PROCEDURES ..................... 1
Alkalinity (Titrimetric) ............................................................................................................... 2
Biochemical Oxygen Demand (BOD5)..................................................................................... 15
Total Residual Chlorine Analysis ............................................................................................. 22
Free Cyanide CN (F)................................................................................................................. 27
Total Cyanide CN (T) ............................................................................................................... 30
Chromium, Total and Hexavalent (Cr 6+ ) ................................................................................ 34
Dissolved Oxygen Analysis (D.O.) .......................................................................................... 39
Fecal Coliform - Membrane Filter Procedure........................................................................... 44
m-ColiBlue24 Method for the Determination of Total Coliforms and E. coli ......................... 54
Hardness, Total (mg/L as CaCO3; Titrimetric, EDTA) ............................................................ 60
pH (Electrometric) .................................................................................................................... 63
Total Phosphorus Analysis ....................................................................................................... 69
Ascorbic Acid Method.............................................................................................................. 69
Settleable Solids........................................................................................................................ 78
Total Suspended Solids (TSS) .................................................................................................. 81
Total Solids ............................................................................................................................... 85
Volatile Acids ........................................................................................................................... 89
Volatile Solids........................................................................................................................... 94
WEST HICKMAN CREEK SOPs.................................................................................................. 1
WH Dissolved Oxygen Field Analysis (D.O.)............................................................................ 2
WH pH (Electrometric)............................................................................................................... 7
WH Total Residual Chlorine Analysis ..................................................................................... 12
WH Fecal Coliform - Membrane Filter Procedure ................................................................... 17
WH Total Phosphorus Analysis................................................................................................ 24
APPENDIX D LABORATORY BENCHSHEETS ................................................................ D-1
d
TB Laboratory QAP
Revision 1
September 16, 2009
FORWARD
The following Quality Assurance Plan (QAP) for the Town Branch Laboratory has been adapted
from the Illinois Water Environment Association Laboratory Committee Model QAP. This
document is intended as guidance only and not a detailed explanation of the accreditation rules.
Periodic review of this document will be conducted and updates will be made as needed to keep
this document up to date and relevant. The Town Branch Laboratory, as part of the LexingtonFayette Urban County Government (LFUCG), prepared this document to serve as a
clearinghouse of information related to the operation of the Laboratory housed at Town Branch
WWTP. The Laboratory serves the:
•
•
•
Town Branch WWTP KPDES Permit No. KY0021491
West Hickman WWTP KPDES Permit No. KY0021504
Blue Sky WWTP
KPDES Permit No. KY0027286
The laboratory is operated in accordance with the Federal Register 40 CFR Part 122, 136, et al.
Primary references include:
•
EPA – 600/4-79-020. Methods for Chemical Analysis of Water and Wastes. U.S.
Environmental Protection Agency; Office of Research and Development, Washington,
DC, 1982.
•
EPA – 600/8-78-017. Microbiological Methods for Monitoring the Environment: Water
and Wastes. U.S. Environmental Protection Agency; Environmental Monitoring and
Support Laboratory, Office of Research and Development, Washington, DC, 1978.
•
Standard Methods for the Examination of Water and Wastewater. APHA-American
Public Health Association. Standard Methods. 21st edition ed.; American Water Works
Association and Water Pollution Control Federation: Washington, DC, 2005.
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Section 1
Quality Policy Statement
1.1
Quality Policy Statement
This Quality Assurance Plan (QAP) provides a written plan of operation for the
laboratory that allows for accuracy, precision, and reliability of laboratory analyses and
that data produced in the laboratory meets or exceeds user requirements. Good
Laboratory Practices are employed to maximize data reliability.
1.2
Specific Objectives
The laboratory will employ methods capable of meeting user's needs for accuracy,
precision, sensitivity, and specificity. Whenever possible, analytical methods used will be
those approved by regulatory or accrediting authorities.
Under the supervision of the Laboratory Management, laboratory staff members will
receive training in quality technology of sufficient depth to perform their assigned duties.
The laboratory will establish a level of quality for routine performance to use as a
baseline from which to measure quality improvement efforts.
Laboratory staff will monitor routine operational performance through analysis of
appropriate quality control solutions and through participation in inter-laboratory testing
programs. The laboratory will provide for corrective actions as necessary
The laboratory will operate in conformance with requirements established by the State of
Kentucky and/or the USEPA.
1.3
QAP Availability
Copies of the laboratory's QAP are available from:
Dr. David J. Price, Ph.D., Laboratory Supervisor
Mrs. La Vada M. Green, QA/QC Manager
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Section 2
Laboratory Organization and Staff Responsibilities
2.1
Laboratory Organization
The Laboratory operates as a department of the Division of Water Quality for the
Lexington-Fayette Urban County Government (LFUCG). The laboratory performs
analyses on samples from the Town Branch WWTP, the West Hickman WWTP, the Blue
Sky WWTP, several industrial pretreatment samples, and samples brought in from other
outside sources. A list of samples and analyses performed are presented in Appendix A.
2.2
Staff Responsibilities
Appendix B contains job descriptions and responsibilities for all laboratory positions.
2.3
Current Personnel
Appendix C lists the laboratory's positions, names of personnel, education, and approved
signatures.
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Section 3
Organizational Charts
3.1
Laboratory's Place in Company and Laboratory Organization
3.2
Laboratory’s Organization
3.2.1 Mrs. La Vada Green - QA/QC Manager
3.2.2 Mr. DiLinh Cao – Laboratory Safety
3.2.3 Mrs. Maria Lundin – Microbiology
3.2.4 Mr. Brian Reynolds – Database administration
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Section 4
Documentation Control and Maintenance Procedures
4.1
Scope
The laboratory maintains document control procedures for its QAP and all standard
operation procedures (SOPs) including analytical procedures and sample preparation
procedures.
4.2
4.3
QAP Documentation Control Procedures
4.2.1
Each page of the QAP includes the title, revision number, effective date, and page
number.
4.2.2
Copies of previous versions of the QAP are archived and kept by laboratory
management and are subject to the record keeping requirements in Section 14.
4.2.3
When minor revisions are made to a section of the QAP, the updated section is
added to the QAP and the previous version is removed and archived for a
minimum of five years.
4.2.4
Archived information shall be available to regulatory agencies.
SOP Documentation Control Procedures
4.3.1
All SOPs are assigned a unique name or code.
4.3.2
Each SOP document contains a revision number.
4.3.3
Each page of a SOP includes its unique name or code, revision number, effective
date, and current page number of total pages in section.
4.3.4 Copies of previous versions of SOPs are archived and kept by laboratory
management for a minimum of five years. These are subject to the record
keeping requirements in Section 14.
4.3.5
4.4
SOPs are available to laboratory personnel as part of a comprehensive lab manual.
New SOPs are added to the lab manual as they are issued. When SOPs are
revised, the revision is added to the lab manual and the previous version is
removed and archived. Likewise, any procedure no longer in effect is removed
from the lab manual and archived for a minimum of five years.
Forms/Data Sheets Documentation Control Procedures
All forms and data sheets prepared by the laboratory display a title, version number, and
effective date.
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Section 5
Laboratory's Approved Signatories
5.1
Approved Signatories
The laboratory's approved signatures with job titles and accreditation position are shown
in Appendix 3. Also, the QAP title page has signed concurrence with appropriate titles of
all responsible parties, including the quality assurance officer and laboratory director.
Laboratory Supervisor
Quality Assurance Officer
Town Branch Superintendent
West Hickman Creek Superintendent
Director, Division of Water Quality
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Section 6
General Quality Control Procedures and Practices
6.1
6.2
6.3
Documentation of Test Methods and Laboratory Practices
6.1.1
All test methods have written standard operating procedures (SOPs). The written
methods describe each procedure and the equipment needed. Each SOP follows a
standard format, which includes additional information on quality control
measures and acceptance criteria for data.
6.1.2
All laboratory practices pertaining to QA/QC as well as laboratory operation
practices other than analytical methods have written standard operating
procedures.
General Quality Control Guidelines
6.2.1
For each test method, the quality control measures described in this QAP are
followed wherever applicable. Additional test method quality control measures
may be implemented, providing they are more stringent than those in this QAP.
6.2.2
All quality control protocol and test procedures are assessed and evaluated on an
on-going basis.
6.2.3
Quality control procedures follow the direction provided in the test method SOP
for evaluation of results; and accept, reject, or qualify sample data based upon the
acceptance criteria specified in the test method. The laboratory establishes the
evaluation procedure and acceptance criteria for a quality control procedure when
not specified by the test method. Whether specified in the test procedure or
established by the laboratory, the evaluation procedures and the acceptance
criteria are documented either in this QAP or in the test method SOP.
6.2.4
If a quality control procedure results in the laboratory rejecting or qualifying
sample data, the problem is investigated, appropriate corrective action
implemented, and the incident documented.
Method Blanks
6.3.1 Method blanks are prepared and analyzed with each batch of environmental
samples and are carried through the entire analytical process.
6.3.2
The method blank is acceptable if it does not contain an analyte of interest at a
concentration greater than the highest of the following: (a) The MDL of the
regulatory limit for that analyte; (b) the MDL of the measured concentration for
that analyte in any environmental sample in the batch; or (c) categorical limits,
such as found in BOD analysis.
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6.3.3
When method blank acceptance criteria are not met, any non-detect results in the
associated batch of environmental samples are reported with qualification.
6.3.4
Section 15, Corrective Action Policies and Procedures, references procedures for
taking corrective actions when blanks do not meet acceptance criteria.
Matrix Spikes
6.4.1
For all test methods performed by the laboratory in which materials suitable for
matrix spiking are available, matrix spikes are performed at a rate of one per
sample set of similar matrix type, per sample extraction or preparation procedure.
6.4.2
The laboratory utilizes the spiking analytes specified in the test method. When the
test method indicates that all method analytes are to be matrix spiked, then the
laboratory spikes all analytes of interest.
6.4.3 Samples are selected on a rotating basis for matrix spike analysis from among
various waste streams, monitoring locations and other applicable locations.
6.5
6.4.4
Procedures used to select samples and analytes for spiking are documented.
6.4.5
Each analytical SOP references quality control criteria to use in determining
discrepancies and accepting data when matrix spikes do not meet acceptance
criteria.
6.4.6
Section 15, Corrective Action Policies and Procedures, references procedures for
taking corrective actions when matrix spikes do not meet acceptance criteria.
Laboratory Control Samples
6.5.1
For each test method, laboratory control samples (LCS) are analyzed at a
minimum of one per batch.
6.5.2
The results of the LCS analyses are used to determine batch acceptance.
6.5.3
Standards for preparing LCS samples are obtained from a second source.
6.5.4
The matrix spike sample may be used as an LCS when the matrix spike
acceptance criteria are as stringent as the LCS acceptance criteria and a LCS has
not been prepared and analyzed for the sample batch.
6.5.5
Each analytical SOP references quality control criteria to use in determining
discrepancies and accepting data when laboratory control samples do not meet
acceptance criteria.
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6.6
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Section 15, Corrective Action Policies and Procedures, references procedures for
taking corrective actions when laboratory control samples do not meet acceptance
criteria.
Matrix Spike Duplicates and Sample Duplicates
6.6.1
Matrix spike duplicates or sample duplicates are performed at a rate of one per 20
or fewer environmental samples per matrix type, per sample extraction, or
preparation procedure.
6.6.2
The sample used for the matrix spike duplicate is the same as used for the matrix
spike.
6.6.3
When sample duplicate analyses are performed, samples are selected on a rotating
basis from among various water samples, wastewater samples, monitoring
locations, and other applicable locations.
6.6.4
All procedures used to select samples for matrix spike duplicate or sample
duplicate analyses are documented.
6.6.5
Each analytical SOP references quality control criteria to use in determining
discrepancies and accepting data when duplicate samples do not meet acceptance
criteria.
6.6.6
Section 15, Corrective Action Policies and Procedures, references procedures for
taking corrective actions when duplicate samples do not meet acceptance criteria.
Quality Control Charts and Tabulations
6.7.1
The laboratory maintains tabulations, quality control charts, or a combination of
tabulations and quality control charts for the results from all quality control
procedures. All calculations for the tabulations and/or control charts are
performed according to Standard Methods, 21th Edition, Part 1020-B.
6.7.2
Separate tabulations and/or control charts are maintained for each test method, for
each matrix, and for each analytical range.
6.7.3
Each tabulation and/or control chart includes the following information:
• Title
• Identification of standard operating procedure
• Name of quality control procedure being tabulated
• Analytical method
• Analyte
• Analyte units of measure
• Matrix
• Fortification concentration
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•
•
•
•
•
•
•
•
6.8
Mean
Standard Deviation
Upper Control Limit (UCL)
Lower Control Limit (LCL)
Upper Warning Limit (UWL)
Lower Warning Limit (LWL)
Date of Analysis
Analyst Identification
Initial Demonstration of Method Performance Studies (IDMP)
6.8.1
6.8.2
Each analyst performs an IDMP study prior to initiation of sample analyses,
unless the IDMP is not applicable to the approved test method, such as: BOD,
CBOD, total suspended solids, total dissolved solids, total volatile solids, total
solids, pH, temperature, dissolved oxygen, or turbidity. The IDMP study is
repeated whenever there is a change in analyst, instrument type, or approved test
method. The following steps are performed for an IDMP study:
•
A quality control (QC) check sample is obtained from an outside source. If not
available, the QC check sample may be prepared by the laboratory using
standards that are prepared separately from the calibration standards by
someone not running the test.
•
The laboratory prepares four aliquots of the QC check samples at the required
method volume to a concentration approximately 10 times the method-stated
or laboratory-calculated MDL.
•
The three aliquots are prepared and analyzed according to the approved test
method.
•
Using the three results, the average recovery and standard deviation are
calculated in the appropriate units for each analyte.
•
For each analyte, the standard deviation and average recovery are compared to
the corresponding acceptance criteria for precision and accuracy in the
approved test method (if applicable) or laboratory-generated acceptance
criteria (if a non-standard method). If standard deviation and average recovery
for all analytes meet the acceptance criteria, the analysis of actual samples
may begin. If any one of the analytes exceeds the acceptance range, the
performance is considered unacceptable for that analyte.
The laboratory management maintains a file to track all current IDMP studies.
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Method Detection Limits (MDL)
6.9.1
MDLs for each analyte of interest are determined by the test method procedure
specified in 40 CFR, Part 136, Appendix B, unless the test method specifies
another procedure for MDL determination, or the determination of an MDL is not
applicable to the test method.
6.9.2 The laboratory analyzes a minimum of seven replicates to determine the MDL
•
•
6.9.3
The laboratory determines MDLs for each approved test method annually; and
when there is a change in instrument type.
6.9.4
The laboratory may, in lieu of the annual determination of the MDL, annually
verify the MDL by the preparation and analysis of a minimum of one matrix spike
sample, spiked at the current MDL. Results are verified by the laboratory
supervisor.
•
•
6.10
If seven replicates are analyzed, the laboratory uses all analytical results
when calculating the MDL.
If the laboratory analyzes more than seven replicates, the laboratory only
excludes analytical results which the laboratory determines are outliers by
utilizing a statistical outlier test.
An MDL is considered verified and acceptable for continued use if results
of the analysis of the matrix spike sample are within the 95% confidence
interval as set forth in Appendix B of 40 CFR, Part 136.
If an MDL cannot be verified, a new MDL is determined.
6.9.5
MDL replicate percent a recovery acceptance criterion is defined by the range of
the percent mean recovery ± 2 times the percent relative standard deviation (%
RSD) found for the seven replicates. If any of the seven replicates fails this
acceptance criterion, then the analyst discards all results and performs another set
of seven replicates.
6.9.6
The spiking concentrations used to determine an MDL are between 1 and 10
times the calculated MDL.
Internal Audits
6.10.1 The laboratory conducts annual internal audits of its operations, QA/QC practices,
and record keeping.
6.10.2 The internal audit is performed by the designated quality assurance officer.
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6.10.3 The results of the internal audit should specify procedures or practices that are not
in compliance with the QAP, and corrective action shall be taken and
documented.
6.10.4 Where the results of an internal audit indicate that the laboratory’s test results are
invalid, the laboratory takes immediate corrective action and immediately
notifies, in writing, the parties that receive the data.
6.11
Analytical Quality Control Standards
All analytical quality control standards that are used are traceable to a National Standard.
6.12
Reagents
Reagents are prepared using reagent grade chemicals or better. Each reagent container
received by the laboratory is documented by marking the container with an adhesive label
and indelible ink. Each marking label indicates the date received, date opened, and any
applicable expiration date.
6.13
Glassware
Glassware used for purposes that may subject it to damage from heat or chemicals is
made of borosilicate glass. Glassware used for volumetric measurement purposes is Class
A rated.
6.14
Laboratory Pure Water
The Town Branch Laboratory uses Nanopure grade water. Tap water is purified through a
series of steps including: coarse filtration, water softening, chlorine removal filtration,
reverse osmosis purification, and 4-cartridge Nano-pure purification process. The nanopure system provides continuous conductivity readings of the of the final product.
Conductivity values range between 17.5 and 18.0 megOhm. Readings below this range
indicate spent cartridges and a need to service the unit. In addition, the resistivity of the
Nanopure water is checked periodically with a calibrated conductivity meter. Resistivity
values of the laboratory pure water are at least 0.5 megOhm/cm at 25°C. Filters are
replaced when the values reach 1.0 megOhm/cm at 25°C.
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Section 7
Verification Practices
7.1
7.2
Performance Evaluation (PE) Testing
7.1.1
Samples are analyzed for this laboratory facility as appropriate for the mandatory
USEPA DMR-QA Laboratory Performance Evaluation Study.
7.1.2
PE samples are analyzed once per year, where appropriate samples are available,
for each test method, each matrix and each analyte included in its scope of
accreditation as required by the provisions in the DMRQA Announcement Letter
(308 Letter) as well as the requirements detailed in the U.S. EPA National
Standards for Water Proficiency Testing Studies Criteria Document (December
30, 1998).
7.1.3
PE samples are processed without any extraordinary care as the results obtained
will be considered typical of the laboratory’s performance. PE samples are treated
as “unknown samples” according to guidelines described in Section 11. All
directions are followed without changing sample preparation, dilution, or analysis.
7.1.4
The laboratory’s personnel do not engage in inter-laboratory communications
regarding PE sample results, or attempt to obtain the true values of the PE
samples prior to reporting at the designated deadline for the PE Study.
7.1.5
All unacceptable results for PE samples are investigated by a standardized
procedure. Appropriate corrective actions are implemented where assignable error
was found. When assignable error is not found, corrective actions are focused on
review of the procedure and improvement of test method execution. The test
procedure is re-validated by successful analysis of a second source standard or
reference material.
Standard Reference Materials
The laboratory uses standards that are traceable to Standard Reference Materials (SRMs),
where available.
7.3
Internal Quality Control Programs
7.4.1
Statistical Process Control is generally used to establish batch acceptance criteria
for analytical test results. The test results for the laboratory control standard are
evaluated annually to set limits for control charts. Warning limits are set at ± 2σ
from the mean recovery, and control limits are set at ± 3σ.
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7.4.2
When a control limit is exceeded, the analyst is required to respond in a manner
that assumes something is wrong with the measurement system. Whenever
possible, a response to an exceeded control limit includes the following:
• Inform the immediate supervisor of the control limit exceedance.
• Stop the analysis of samples, if possible.
• Conduct a systematic investigation as soon as possible to locate the source
of the problem.
• Take appropriate corrective action when a problem is located.
• Rerun samples to the last good laboratory control standard whenever
possible.
• Document the control limit event, including the details of the occurrence,
whether a problem was detected, and any corrective actions taken.
• Maintain a state of increased vigilance.
7.4.3
Warning limit trend exceedance occurs when two or more consecutive results for
the laboratory control standard exceed the warning limit. A general response to a
trend exceedance includes the following.
• Inform the immediate supervisor of the warning limit trend exceedance.
• Conduct a systematic investigation as soon as possible to locate the source
of the problem.
• Take appropriate corrective action when a problem is located.
• Document warning limit exceedance, including the details of the
occurrence, whether a problem was detected, and any corrective actions
taken.
7.4.4
The laboratory has a written response plan for the analyst to follow in the event of
a failure of a laboratory control standard or other criteria. This plan is found in
Section 15, Corrective Action Policies and Procedures.
7.4.5
SOPs for test methods and lab procedures are reviewed on an annual basis. The
designated Quality Assurance Officer and Laboratory Supervisor performs
reviews.
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Section 8
Equipment Procedures for Calibration, Verifications, and Maintenance
8.1
Equipment Calibration
8.1.1
An initial calibration is performed on instrumentation and equipment as specified
in the test method. Calibration standards are traceable to a National Standard,
where available.
8.1.2
The procedures for calibration verification and maintenance are found in the
analytical method SOPs or sample preparation SOPs. Manufacturers operation
manuals may be referenced in the method SOPs when they are the source for
calibration or maintenance procedures.
8.1.3
Documentation is maintained for all maintenance, calibration and instrument
operation activities. All defective equipment is removed from service and is not
returned to operation until repaired and shown by calibration, certification or test
to perform satisfactorily.
8.1.4
An adequate number of standards are used to define the calibration curve. The
test method SOP states if the calibration curve is linear or non-linear.
8.1.5
If the test method does not state the number of calibration standards to use, the
laboratory will use a minimum of 7 concentrations to create the calibration curve.
The range should be within the linear range of the curve, and should correspond
to values typical for the samples.
8.1.6
Unless specified by the test method, the lowest calibration standard is set at 1 to
15 times the MDL whenever sample results will be used in a decision related to
the determination of a non-occurrence of an analyte or a non-detect the MDL of
an analyte.
8.1.7
All sample results for test methods utilizing a calibration curve are reported
within the highest calibration standard, or within the linear dynamic range where
the test method requires determination of the linear dynamic range.
8.1.8
Further guidance regarding calibration may be found in Section 186.155 of
Illinois regulations Title 35: Environmental Protection, Subtitle A: General
Provisions, Chapter II: Environmental Protection Agency, Part 186: Accreditation
of Laboratories for Drinking Water, Wastewater, and Hazardous Waste Analysis.
Copies of this document may be obtained from the laboratory supervisor or the
quality assurance officer.
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Balances
The laboratory’s analytical balance has a sensitivity of 0.1 mg. All analytical balances are
placed on a stabilizing slab base. Each balance is checked daily with two or more ASTM
type 1 weights, which cover the effective range of the balance’s use. All balances are
serviced and calibrated at least annually by a qualified service representative. The service
representative issues the laboratory a certificate of calibration with weights traceable to
National Standards.
8.3
pH Meters
All pH meters have an accuracy of at least ± 0.1 pH units and provide for temperature
correction of pH measurements. Daily calibrations are performed with a minimum of
three standardization buffers as specified in SOP-pH. The pH of the standardization
buffers are 4.0, 7.0 and 10.0.
8.4
Conductivity Meters
A conductivity meter is maintained with an error not exceeding 1% or one
micromhos/cm, whichever is greater. The conductivity meter is calibrated before each
use with a standard that reflects the sample conductivity.
8.5
8.6
Thermometers
8.5.1
The laboratory has certified thermometers traceable to National Standards, with 1
degree centigrade or finer subdivisions and a range which spans the various
requirements of the analytical methods, equipment temperature monitoring, and
checking for thermal preservation. These traceable thermometers are recalibrated
a minimum of every five years. The laboratory maintains Certificates of
Calibration that identify traceability of the calibration to National Standards.
8.5.2
All other thermometers are calibrated against thermometers traceable to National
Standards. Liquid-in-glass and digital thermometers are calibrated annually. Metal
and continuously monitoring thermometers are calibrated at least quarterly.
Calibration factors are employed based upon the most recent calibration.
Refrigerators
8.6.1
Each refrigerator is uniquely identified and provided with a uniquely identified
thermometer graduated in increments no larger than 1 degree centigrade.
Thermometer readings are monitored and recorded each day the laboratory is in
operation. The monitoring logs include refrigerator and thermometer
identification, date, time, temperature, initials of the responsible person, and the
acceptable temperature range.
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8.6.2
The following table lists the refrigerators that are used in the Town Branch
Laboratory.
Description
Room Temperature Storage Unit
Microbiology Station
Pretreatment Sample Storage
Town Branch Sample Storage
West Hickman Sample Storage
Chemical Storage
8.6.3
8.7
Unique Identifier
Room Temp Storage Unit
Refrigerator #1
Refrigerator #2
Refrigerator #3
Refrigerator #4
Refrigerator #6
Range
10 - 30o C
4.0 ± 2.0o C
4.0 ± 2.0o C
4.0 ± 2.0o C
4.0 ± 2.0o C
4.0 ± 2.0o C
Samples which require thermal preservation are stored under refrigeration which
is ± 2o C of the specified preservation temperature, unless method specific criteria
exist. For samples with a specified storage temperature of 4o C, storage
temperatures of 4.0 ± 2.0o C are acceptable.
Incubators
8.7.1
Each incubator is uniquely identified and provided with a uniquely identified
thermometer graduated in increments no larger than 1o C. Thermometer readings
are monitored and recorded each day the laboratory is in operation. The
monitoring logs include incubator and thermometer identification, date, time,
temperature, initials of the responsible person, and the acceptable temperature
range.
8.7.2
The following table lists the incubators that are used in the Town Branch
Laboratory.
Description
BOD Incubator
BOD Incubator
Coliform Incubator Bath
Coliform Incubator Bath
Coliform Incubator Bath
8.8
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Unique Identifier
BOD Incubator #1
BOD Incubator #2
Precision #1
Precision #2
Precision #3
Range
20.0 ± 1.0o C
20.0 ± 1.0o C
44.5 ± 0.2o C
44.5 ± 0.2o C
35.0 ± 0.2o C
Ovens
8.8.1
Each oven is uniquely identified and provided with a uniquely identified
thermometer, graduated in increments no larger than 10o C for muffle furnaces
and 1o C increments for oven and warm air incubators. Temperatures are recorded
daily. The monitoring logs include; oven and thermometer identification, date,
time, temperature, initials of the responsible person, and the acceptable
temperature range.
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8.8.2
The following table lists the ovens that are used in the Town Branch Laboratory.
Description
Drying Oven
Iso-temperature Oven
Muffle Furnace
8.9
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Unique Identifier
Drying Oven #1
Iso Temp Oven
Muffle Furnace
Comments
103 – 105o C
Adjustable
Adjustable
Other Laboratory Equipment
8.9.1
The laboratory monitors and controls method specific temperature requirements
for heating blocks and water baths, etc. The laboratory also maintains
documentation of the results.
8.9.2
Autopipetors and dilutors of sufficient accuracy are used for some applications.
The autopipetors (repipettors) are serviced and calibrated annually by a qualified
service representative. The service representative issues the laboratory a
certificate of calibration for each unit and labels each unit to indicate
service/calibration. Logs of these checks are maintained for each device.
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Section 9
Test Methods and SOPs
9.1
Scope
The Standard Operating Procedures section describes all procedures currently in use
by the laboratory. Included in this section are each method's SOP number, document
name, source, and whether it is listed in 40 CFR Part 136 as an approved method.
9.2
Standard Operating Procedures (SOPs)
The SOPs are incorporated in this Quality Assurance Plan and may be obtained from
the laboratory management. In addition, each laboratory staff member is issued a
manual containing all current SOPs. A laboratory copy is used for updates and
changes between revisions.
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Section 10
The Laboratory’s Physical Facilities Including Services and Resources
10.1
Building
The Town Branch Laboratory, operated under the Lexington-Fayette Urban County
Government, is located in the Town Branch Administration building, 301 Lisle
Industrial Avenue, Lexington, Kentucky. The Town Branch Laboratory performs
routine monitoring for Town Branch Wastewater Treatment Plant, West Hickman
WWTP, and Blue Sky WWTP, several industrial pretreatment samples, and samples
brought in from other outside sources. The Administration building housing the
laboratory was constructed in 1970. Currently, the laboratory and offices occupy
5,735 square feet of floor space. The laboratory originally occupied 2,370 square feet
of floor space. In 2004, the lab was expanded to include an additional 3,365 square
feet of floor space. The new addition includes additional bench space and cabinets. A
schematic layout of the laboratory showing current space utilization is shown in
Appendix 5.
10.2
10.3
Access and Security Measures
10.2.1
Entrances to the Town Brach WWTP are located at the north-east and
south ends of the plant. Automatic gates are open during normal operating
hours. Magnetic access cards are required for entry during off hours.
10.2.2
Any visitors to the Town Branch WWTP and Laboratory are required to
sign in with Reception in the Administrative Building.
10.2.3
Normal operating hours for the laboratory are 7:30 AM to 4:00 PM,
Monday through Friday. A Laboratory Technician is present 7:30 AM to
4:00 PM on Saturdays.
Building Services
10.3.1
Air Temperature and Quality - The laboratory’s heating and airconditioning are generally to control laboratory temperatures between 68
and 75o F and temperature fluctuations rarely exceed more than 4 degrees
per hour.
10.3.2
Electricity - 110 and 220 volt electrical service is provided throughout the
laboratory.
10.3.3
Illumination - Lighting in most areas and all work areas is provided by
fluorescent lighting. Emergency lighting is located throughout the lab.
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Safety
10.4.1
Fire Safety – Fire alarms are located throughout the building. Fire
extinguishers are prominently displayed throughout the lab and all
laboratory personnel are trained yearly in the use and safe handling of the
extinguishers.
10.4.2
Other Safety Equipment – The Town Branch lab maintains a fully stocked
first aid kit, two eye-wash and shower stations. All safety equipment is
inspected on a monthly basis. The lab has a total of 8 chemical hoods,
which are inspected and serviced yearly. Chemical neutralization and spill
control kits, and acid and flammable storage cabinets are located in the
chemical storage room. Current MSDS sheets are maintained for all
chemicals used in the lab.
Computer Resources Including Equipment and Software
The LFUCG IT department is responsible for all computers and software used in the
laboratory. A total of 5 PC computers are located throughout the laboratory. All
computers have access to the internet via the City’s server. Password protections
restricts access to lab data to lab personnel only. All data is viewable as a “read-only”
file. In addition to the Excel-based database, all laboratory computers can access the
HACH Water Information Management Solution (WIMS) database which houses all
laboratory information.
10.6
Laboratory Work Areas
Several sections in the laboratory have been reserved for specific analyses. These
sections include: sample receiving, microbiology, BOD preparation/analysis, wetchemistry, total residual chlorine analysis, total cyanide digestion, solids
processing/weighing, spectrophotometric methods (i.e. HACH TNT+ methods), water
purification. Employees are routinely rotated through the different procedures.
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Section 11
Procedures for Reviewing New Work
11.1
Scope
Laboratory management reviews all new work to ensure the laboratory is able to perform
the additional task(s) in a timely and accurate manner. Laboratory management consults,
if necessary, with sampling personnel and end data users to ensure sample integrity and
data quality.
11.2
Unknown Samples
On occasions, the Laboratory will receive and analyze samples from industrial
pretreatment sources or samples brought in from other outside sources. These samples are
considered “unknown samples” for which duplicate sub-samples are processed and
analyzed. All QA/QC requirements apply to the unknown samples.
11.3
Contract Analyses
The laboratory does not perform contract analyses.
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Section 12
Sample Acceptance and Receipt Policy
12.1
Scope
Sampling is critical to producing data representative of conditions that occur at the
sampling location. This section of the QAP constitutes the laboratory's written sample
acceptance policy. It details the conditions under which the laboratory will accept
samples. It is readily available to all laboratory staff and sample collectors.
12.2
Sample Acceptance Requirements
12.2.1 The Plant Operator confirms the appropriate sampling plan in order to verify that
sampling techniques are consistent with the intended use of the data. Sampling
equipment is selected and pre-cleaned to preserve sample integrity and eliminate
contamination.
12.2.2 Accepted samples are logged upon arrival and checked for sampling preservation
and holding times.
12.2.3 Indelible ink is used for all written documentation associated with samples.
Corrections to sampling documents are made by a single line strikeout. The
corrected entry is written above the strikeout and initialed by the person making
the correction.
12.2.4 Samples must be received with complete documentation in the form of a Chain of
Custody (COC) record including sample identification, location, date and time of
collection, collector’s name, sample type, person receiving sample, and any
special remarks.
12.2.5 COC Records
The different COC records used at Town Branch Laboratory are:
1) Town Branch 24-hour composite samples with COC record (delivered to the
Town Branch Lab by the night Operator at midnight the night before).
2) Town Branch grab samples with COC record (delivered to the Town Branch
Lab each morning by the on-duty Operator).
3) Town Branch solids grab samples and composite samples with COC record
(delivered to the Town Branch Lab each morning by the Solids Operator).
4) West Hickman grab and composite samples are on one COC record
(delivered to the Town Branch Lab each morning by the West Hickman onduty Operator).
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5) Blue Sky grab and composite samples are on one COC record (delivered to
Town Branch Lab every Thursday by the Blue Sky on-duty Operator).
6) Illicit Discharge Detection and Elimination (IDDE) grab samples with COC
record (delivered to Town Branch lab by the Environmental Inspector).
7) Industrial waste 24-hour composite samples COC record (delivered to the
Town Branch Lab by the Environmental Inspector).
12.2.6 COC Checklist
1) Operator obtains COC record.
2) Operator collects samples.
3) On COC record operator records:
• date sampling started
• time sampling started
• date sampling stopped
• time sampling stopped
• sampling method
• sample type
• sample size
• type of preservative
• analysis requested
• sampler’s signature
4) Operator delivers samples to Town Branch Lab.
5) Operator signs the COC sheet on the line “Relinquished By (Representing
Operators) (Signature)”.
6) Operator writes the date and time of relinquishment on the same line as
noted above in (5).
7) Operator leaves the COC sheet with a Laboratory Analyst who then signs
the sheet on the line “Received in the Laboratory By (Signature)”.
• On the same line the Laboratory Analyst writes the date and time the
samples were received in the lab.
12.2.7
Samples must be received in the sample container required by the test method
and properly labeled.
12.2.8
Each sample container must have a unique identification.
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Sample preservation must comply with the requirements of the approved test
method.
12.2.10 Sample volume must be sufficient to perform the necessary analyses.
12.2.11 Samples must be received within the time specified by the test method. Upon
receiving a sample, the laboratory will determine if the analysis can be
performed within the allotted holding time.
12.2.12 Bypass samples and Town Branch 24-hour composite samples are collected the
night before analysis and are delivered to Town Branch Laboratory and kept in
a secure location until analysis. The chain of custody (COC) form is stored with
the samples.
12.2.13 Town Branch grab samples, West Hickman 24-hour composite samples, Blue
Sky 24-hour composite samples, and Industrial Waste 24-hour composite
samples are brought to the Laboratory Analyst during operating hours.
12.3
Verification of Preservation
12.3.1 Samples are examined for method preservation and proper documentation.
12.3.2 During 24 hour composition, samples are examined daily for thermal
preservation. Thermal preservation will be acceptable if:
•
Temperature is either within ± 2° C or at the method’s specified range. For
samples with a specified temperature of 4° C, a temperature of 2.0 to 6.0° C is
acceptable.
•
Samples have been hand delivered to the laboratory within 6 hours of
collection and there is evidence that the chilling process has begun, such as
arrival on ice.
12.3.3 Town Branch Lab refrigerated storage unit temperatures are checked and
documented daily. Temperature must be 4º C ± 2° C.
12.3.4 Samples are examined for chemical preservation upon receipt, or prior to
additional sample preparation or analysis.
12.4
Actions for Deviations from the Lab's Sample Acceptance Requirements
12.4.1 Deviations from the sample acceptance policy is brought to the attention of the
Laboratory Analyst and documented at the time of sample log-in.
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12.4.2 Whenever possible, the laboratory proceeds to correct errors in sample
documentation. The person responsible for the error is contacted so that a
deviation from the sample acceptance policy can be avoided.
12.4.3 If a sample does not meet all of the sample acceptance criteria:
• Retain correspondence and records of information concerning the final
disposition of rejected samples.
• Contact client from which the laboratory received samples.
12.5
Laboratory Log of Received Samples
Each sample received is logged with the following information:
•
•
•
•
•
•
•
•
•
•
date and time of laboratory receipt of sample
sample collection date
whether sample is composite or grab
unique laboratory identification code
sample collection point
requested analyses
signature or initials of data logger
comments resulting from inspection for acceptance or rejection
sample collector
person receiving sample
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Section 13
Sample Tracking and Storage Procedures
13.1
Scope
A clear sample tracking record allows any questions concerning sample integrity to be
answered during any step of the process. Samples are stored in areas which isolate
samples sufficiently to prevent cross-contamination with other samples, reagents or
standards. Procedures for sample handling and tracking may vary, depending upon
sample origin. Sample integrity and reliable test data are ensured when valid laboratory
procedures are established and strictly followed.
13.2
Sample Tracking Record
13.2.1 Sample logs, bench sheets, and preparatory records are clearly marked to identify
all personnel associated with each sample.
13.2.2 Chain of custody forms or bench sheets must be completed to include use of
preservation and sample containers required by approved test method.
13.2.3 No samples are accepted without proper documentation. Samples are checked to
insure that the container is compatible with the intended analysis.
13.3
Proper Storage and Avoidance of Cross-contamination of Samples
13.3.1 Storage facilities are provided for samples to prevent cross-contamination and are
consistent with sample preservation requirements of the method.
13.3.2 Samples are stored away from all standards, reagents, food and other potentially
contaminating sources.
13.4
Security of Samples
The laboratory stores all samples within the confines of the laboratory and limits access
to authorized laboratory personnel only.
13.5
Sample Disposal
If possible, samples and aliquots are kept to the end of the maximum permitted holding
time in the event that re-analysis of the sample is required. Proper temperature and
holding times must be observed for re-analysis. Analyzed samples or samples with
expired holding times are disposed of properly.
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Section 14
Record Keeping, Data Review and Reporting Procedures
14.1
Scope
The record keeping system allows historical reconstruction of all laboratory activities that
produce the resultant sample analytical data. The laboratory maintains complete sample
tracking records. Documentation of laboratory activities includes inter-laboratory
transfers of samples and sample extracts. The laboratory issues sample data or sample
result reports accurately and in a manner that is understandable to the recipient.
14.2
Records Maintained by the Laboratory
14.2.1 The laboratory retains records related to all procedures and activities to which a
sample is subjected. These records include:
1)
Identity of personnel involved in sampling, preparation and testing.
2)
Sample preservation, including sample container and compliance with
holding times.
3)
Sample identification number, receipt, log-in, acceptance or rejection.
4)
Sample storage and tracking, including shipping receipts, transmittal forms,
and internal routing and assignment records.
5)
Sample preparation including: cleanup and separation procedures, sample
identification codes, volumes, weights, instrument printouts, and
calculations.
6)
Sample analysis.
7)
Equipment receipt, use, specification, operating conditions and preventative
maintenance.
8)
Calculations and statistical formulas used by the laboratory, including
written procedures for calculations, raw data and supporting information
used for each calculation, correct use of significant figures, and
identification of the least precise step in accord with limitations of the
measurement system.
9)
Procedure to verify that the reported data is free from transcription and
calculation errors.
10) Data handling, including reduction, review confirmation, interpretation,
assessment or validation, and reporting.
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11) QC measurements, including procedure to select samples on which to
perform QC measurements, and assessment of method performance.
14.2.2 Other records retained by the laboratory include:
1)
2)
3)
4)
5)
6)
14.3
All original raw data, whether hard copy of electronic, for calibrations,
analyst’s worksheets, and data output from instruments or equipment.
Copies of final reports.
Archived SOPs.
Correspondence with sample submitters.
Corrective action reports, audits and audit responses.
PE sample results and raw data.
Records of Standards and Analytical Reagents
14.3.1 Traceability of standards - The laboratory verifies that standards are traceable to
National Standards. If traceability is not possible, the laboratory demonstrates, by
appropriate means (i.e., analyses of PE samples) that the instrumentation and
equipment is properly calibrated.
14.3.2 Receipt and use of reagents and standards - The laboratory retains records of the
origin, purity and traceability of all reagents and standards. These records also
include the date of receipt, storage conditions, the date of opening and an
expiration date.
14.3.3 Traceability of working and intermediate standards - The laboratory maintains
records of traceability from working and intermediate standards to purchased
stock standards or neat compounds which include the date or preparation and
preparer’s initials.
14.3.4 Identification of prepared reagents and standards - All prepared reagents and
standards are labeled with the identity the reagent or standard, concentration,
preparation date, and preparer’s initials.
14.3.5 Records of Instrument or Equipment Calibrations
14.3.6 The laboratory documents and maintains calibration procedures that establish
calibration frequency and calibration acceptance criteria.
14.3.7 All graphs of calibration curves have descriptive titles, labeled axes, and date of
calibration, time of calibration, test method, analytic, standard concentrations,
instrument response and the calibration curve.
14.3.8 All calibrations record the equation of the calibration curve and the correlation
coefficient.
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Computerized Data Storage
14.4.1 The laboratory establishes and implements procedures for protecting the integrity
of the data, including data protection procedures during data entry and capture,
data storage, data transmission and data processing. Data is protected with lab
password access only.
14.4.2 The laboratory provides procedures for the maintenance of the security of the
data, including the prevention of unauthorized access to data, and the
unauthorized change of computer records.
14.4.3 The laboratory maintains hard copy and/or writes protected backup copies of
computer generated records.
14.5
Administrative Records
14.5.1 Personnel qualifications - The laboratory maintains records of personnel
qualifications, education, experience and training.
14.5.2 IDMP studies - The laboratory maintains records of IDMP studies and any
required repetitions of the IDMP for each analyst.
14.5.3 Initials and signatures - The laboratory maintains a log of names, initials and
signatures for all individuals who are responsible for signing or initialing any
laboratory record.
14.6
Laboratory Record Entries and Change of Entries
14.6.1 Laboratory personnel sign or initial (all three initials) all record entries. The
reason for the signature or initials is clearly indicated in the records, including,
but not limited to: sampled by, prepared by, and reviewed by.
14.6.2 Permanent ink is used for manually recorded data.
14.6.3 Corrections of manually recorded data are made by one line through the error.
The correction is initialed and dated. Corrections are not made by erasure or
White Out™.
14.6.4 Electronically maintained records are kept in such a fashion as to indicate any
change in the record.
14.6.5 After the laboratory delivers its sample data and results to client, the laboratory
will only correct, add or delete information from the report when it supports those
actions by supplementary documentation. Any supplemental report clearly
identifies there purpose and contains all reporting requirements.
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14.6.6 Facilities with in-house laboratories which provide data to regulatory agencies
must include all items in Section 14.10 with their reports if so required by the
regulatory agency.
14.7
Record Retention
14.7.1 All records are retained for a minimum of five years. They include information
pertaining to environmental analyses, performance testing, obsolete or replaced
procedures, and supplies for tests, support services, and laboratory accreditation.
14.7.2 Access to archived records is documented with an access log. All records are
protected against loss of deterioration including fire, theft, and electromagnetic
deterioration in the case of electronic records.
14.8
Raw Data Associated with Sample Analysis
All raw data associated with sample analyses (i.e., calibration curves, strip charts, tabular
printouts, computer data files, analytical notebooks, and run logs) include the following:
1)
2)
3)
4)
5)
6)
7)
14.9
Laboratory sample number.
Date of analysis.
Type of analysis.
Instrument identification.
Instrument operating conditions (or reference to such information).
All calculations automated or manual to which the sample data is subjected.
Analysts’ and/or technician’s initials or signature.
Test Data Reports
Laboratory sample data or sample result reports include the following:
1) Report title such as "laboratory results".
2) A unique identification of the report such as serial number.
3) Description and identification of samples.
4) Date of sample receipt, sample collection and sample analysis (time of
sample preparation and analysis if the required holding time for either
activity is less than or equal to 48 hours).
5) Characterization and condition of sample, where appropriate.
6) Reference to sampling procedure, where applicable.
7) Test method utilized.
8) Sample results with any failures or deviations from approved test methods
or QC criteria identified, such as data qualifiers.
9) Description of the calculations or operations performed on the data, a
summary and analysis of the data.
10) Identification of the reporting units (such as mg/L or mg/kg).
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11) Identification of any results not generated by the laboratory preparing the
data report and identification of the laboratory from which such results were
obtained.
12) A statement that the report shall not be reproduced, except in full, without
the written approval of the laboratory.
13) A statement that samples results relate only to the analytes of interest tested
or to the sample as received by the laboratory.
14) Additional information may be required for sample data reports submitted to
a regulatory authority.
The laboratory certifies that the sample results meet all the requirements of any
environmental laboratory program for which it maintains accreditation.
14.10 Electronic Data Transfer
Electronic data transfer (i.e. fax, email attachments, CD, flash drives) are documented by
hard copies.
14.11 Sample Disposal Records
Ordinarily, the laboratory does not need to maintain records for disposal of samples. For
a record of possible litigation samples and other samples that the laboratory supervisor
wishes to retain, such records will be catalogued by date of disposal and person
responsible. Samples may be disposed of by sample depletion, sample returned to
submitter, sample washed down sink, and sample manifested to a hazardous waste
facility. Any correspondence concerning sample disposal must be retained. Additional
information on confidentiality is presented in Section 18.
14.12 Waste Disposal
Waste collection, storage, recycling, and disposal procedures and policies are part of
method SOPs. Where disposal procedures and policies are included as part of a test
method, the test method disposal practices are strictly followed.
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Section 15
Corrective Action Policies and Procedures
15.1
Scope
The following policies and procedures are used when any analysis or reporting
discrepancies are detected, or when any deviation from the policies and procedures in this
manual occur.
15.2
Identification of Discrepancies
15.2.1 Discrepancies or deviations shall be defined as, but not limited to, any of the
following:
1) quality control sample results outside control limits
2) reporting sample results in wrong units
3) using unapproved analytical procedures
4) data which appears to be erroneous to current trends
15.2.2 Each analytical procedure SOP references quality control criteria to use in
determining discrepancies and accepting data.
15.3
Staff Responsible for Investigation and Corrective Action
15.3.1 Any of the Laboratory staff may detect discrepancies. Once a discrepancy is
detected, it is reported to laboratory supervisor. Laboratory management or staff
may investigate any discrepancies. Data that has discrepancies should not be
recorded and should be reviewed.
15.3.2 Once a discrepancy is established, staff should proceed with one or more of the
following:
1) re-run samples, if available and holding time permits
2) document results as invalid and note on any reports
3) investigate discrepancy; document cause and corrective action taken and
include with data
15.3.3 Staff responsible for investigating the discrepancy shall review sample and quality
control results, integrity of quality control samples, and technique. If a quality
control result does not meet method or laboratory criteria, it shall be documented
on the analysis bench sheet and quality control chart. Staff should also review
sampling procedures and preservation, washing of glassware, and any sources of
contamination to samples or quality control standards.
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15.3.4 The Laboratory staff performs corrective actions to eliminate any of the above
mentioned possible causes for data discrepancies. Examples of corrective actions
are: review of proper procedures, quality control standard preparation, changes to
procedures or sampling protocol, and improving analyst technique. If the
investigation cannot determine a known cause for invalid results, retraining and
procedure review are the appropriate corrective actions.
15.3.4 For each data discrepancy event, the investigation and corrective action shall be
documented.
15.4
Documentation and Review of Corrective Action
Laboratory management reviews raw data, reports, quality control data, any
discrepancies, and corrective actions on a regular basis. This review helps establish any
recurring problems that require further investigation and action.
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Section 16
Procedures for Permitting Departures from Documented Policies and Procedures
16.1
Scope
Departures from documented policies and procedures may include, but not limited to:
• use of analytical methodology which is not KYDEP or U.S. EPA approved
• use of samples which are not properly sampled or preserved for intended purpose
• analysis of samples outside the required holding times
• deviations from analytical or other laboratory SOPs
• deviations from standard QC practices
• use of alternative calibration practices
• reporting data which is not compliant with KYDEP or U.S. EPA analytical or QC
requirements
16.2
Requests for and Approval of Departures
Requests for allowing departure from documented policies and procedures are directed to
the Laboratory Supervisor. The Laboratory Supervisor shall evaluate the reasons for such
departures. The Quality Assurance officer is consulted for all matters that may affect the
quality of analytical data. Written approval by the Laboratory Supervisor is required
before such departures are made. The Laboratory Supervisor documents and files all
departures and their duration. Permanent departures are documented by changes in the
established policies and procedures.
16.3
Use of Analytical Methods Not Approved by KYDEP or U.S. EPA
When analytical methods are used which are not approved by KYDEP or U.S. EPA,
SOPs are used to document the actual procedures performed as well as the source(s) of
the method. The methods used to determine the criteria for IDMP are established and
documented prior to use of the method. Quality assurance and control procedures are
included in the method.
16.4
Reporting of Non-compliant Data
Data which is produced from samples that do not meet the regulatory sampling, holding,
or preservation requirements, or which is produced using methods that deviate from
EPA-required analytical or QC requirements, is reported only when accompanied by a
statement that clearly indicates that the data may not be used for regulatory compliance
purposes.
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Section 17
Procedures for Dealing with Complaints
17.1
Scope
The Town Branch Laboratory has specific procedures for dealing with complaints from
clients or other parties.
17.2
Complaint Receipt Procedures
17.2.1 Document the initial complaint with the following information:
• Name of person with complaint
• Company name
• Phone number
• Sample Identification
• Nature of complaint
17.2.2 Inform person with complaint that it will be investigated promptly. Also inform
him of the estimated time or date for a response to the complaint.
17.3
Complaint Investigation Procedures
17.3.1 Review the following items regarding the sample or analysis in question with
laboratory staff:
• Receipt of sample
• Internal Chain of Custody
• Analyses performed
• Analysis methods
• Analytical results
• Calibration check
• Quality control results
• Calculations
• Unit conversions
17.3.2 Document whether any discrepancies were revealed during the review, especially
discrepancies which address specific complaint.
17.4
Complaint Response Procedures
17.4.1 Laboratory management will contact the person with complaint and reveal
findings of investigation. If necessary, a corrected report will be issued.
17.4.2 File the documented complaint investigation for use during the annual review of
the Quality Assurance Plan. If any corrective actions are required, reference
Section 15 of this QAP manual, “Corrective Action Policies and Procedures.”
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Section 18
Procedures for Protecting Confidentiality and Proprietary Rights
18.1
Procedure for Protecting Confidentiality
The Town Branch Laboratory must protect confidentiality and proprietary rights.
Laboratory employees are instructed not to divulge any information that may involve
issues of confidentiality or proprietary without the approval of the Laboratory Supervisor.
18.2
Examples of Confidential Data
Some examples of confidential data include:
• Effluent and/or Permit data.
• Analytical results for Plant operation samples.
• Analytical results for samples obtained from industrial dischargers.
• Results obtained for the PE.
• Sample results that may be integral to on-going or possible litigation.
18.3
Procedure for Open-Records Requests
Requests for open-records must be made in writing to the Laboratory Supervisor. The
requests will be reviewed by the Laboratory Supervisor and require final authorization by
the Division Director before release of records. The request and released records will be
filed by the Laboratory Supervisor.
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Section 19
Procedures for Internal Audits
19.1
Annual Internal Audit
The Quality Assurance Officer and/or Laboratory Supervisor conducts an annual internal
audit of the laboratory.
19.1.1. This auditor conducts a systematic audit of technical activities from previously
prepared checklists.
19.1.2. The auditor determines whether the quality assurance practices and other
laboratory procedures described or referenced in the QAP have been
implemented.
19.1.3. The auditor then prepares a written report which includes copies of the check-lists
and a list of all noted deficiencies.
19.2
Corrective Actions Regarding the Annual Internal Audit
Laboratory management takes appropriate corrective actions in response to the internal
audit, including a written response plan which covers:
• Completed corrective actions
• Planned corrective actions
• Implementation schedule for planned corrective actions.
19.3
Notification of Invalid Data
In the event that the results of the internal audit indicate that the laboratory’s test results
are invalid, immediate corrective action is taken. All persons are notified who received
the invalid data.
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Section 20
Annual Review of the Quality Assurance Plan (QAP)
20.1
Scope
The QAP is reviewed annually. The Laboratory Supervisor conducts an internal review
with the Quality Assurance Officer. The laboratory, in its review, determines whether
requirements of the QAP have been adequately addressed, the results of the annual
review must be documented.
20.2
Correcting Deficiencies in the QAP
If the Laboratory Supervisor and Quality Assurance Officer find deficiencies, the QAP is
revised as necessary. The revised QAP is then issued with the appropriate version number
and implemented at the agreed upon effective date. All copies of the preceding version
are replaced with the new version.
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Section 21
Training and Personnel Requirements
21.1
Scope and Training Objectives
All personnel involved in laboratory analysis or quality assurance/quality control shall
have sufficient training to allow for the analysis and reporting of complete, high quality
data in compliance with the procedures of this Comprehensive Quality Assurance Plan.
Laboratory Management is responsible for ensuring the required training is made
available.
21.2
Training Required for Laboratory Supervisor
21.2.1 The Laboratory Supervisor shall hold a bachelor’s degree in natural or physical
sciences or have completed enough course work in chemistry to equal a major in
chemistry.
21.2.2 The Laboratory Supervisor shall have a minimum of one year’s experience in
analyses pertaining to the applicable fields of testing.
21.3
Training Required for Quality Assurance Officer
21.3.1 The QA Officer shall hold a bachelor's degree in natural or physical sciences or
have completed enough course work in chemistry to equal a major in chemistry.
21.3.2 The QA Officer shall have a minimum of one year's experience as an analyst in a
laboratory.
21.3.3 The QA Officer shall have documented training in quality assurance and quality
control.
21.4
Training Required for Technicians
21.4.1 Technicians shall hold a bachelor's degree in natural or physical sciences or have
completed enough course work to equal a major in chemistry.
21.4.2 Technicians shall have a minimum of one year's experience in the analyses
pertaining to the applicable fields of testing.
21.4.3 Technicians shall meet the instrument training requirements specified in Section
21.9.
21.4.4 After completing training, technicians shall perform an Initial Demonstration of
Method Performance (IDMP) study for each analysis as specified in Section 6.8.
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21.4.5 The laboratory shall have on file documentation indicating the analyst's
acceptable performance on a blind sample at least once per year and a
certification that the analyst has read, understood and agreed to perform the most
recent version of the standard operating procedure.
21.5
Additional Training for Technicians
Additional training, provided by the Laboratory Management, involves participation in
the Kentucky Laboratory Analyst (KLA) training, testing, and certification for laboratory
technicians. The KLA was created under the KY Water and Wastewater Operators'
Association (KWWOA) for individuals employed in the water quality field. A total of 4
class certifications are available for Wastewater Laboratory Analysts.
21.6
Substituting Experience or Education to Meet Training Requirements
A person may serve as Laboratory Supervisor, Quality Assurance Officer, or Technician
when that person does not meet the training, educational or experience requirements for
the position. In such cases the laboratory shall submit written justification explaining why
a person should serve in a particular position. This written justification shall take into
account the following factors:
• Experience as an offset for education requirements.
• Education as an offset for experience requirements.
21.7
Instrument Training Requirements
Analysts and technicians must meet either of the following requirements for analyses
performed utilizing specialized laboratory instrumentation (i.e., Atomic Absorption):
• The technician shall have satisfactorily completed a minimum of four hours training
that is offered by the equipment manufacturer, a professional organization, a
university, or qualified training facility.
• The technician shall have served a two week period of apprenticeship under an
experienced staff member.
21.7
Training Records
Laboratory management maintains employee training records. The records are updated
periodically for each employee receiving training.
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Section 22
Glossary
Acid - An inorganic or organic compound that (a) reacts with metals to yield hydrogen; (b) reacts
with a base to form a salt; (c) dissociates in water to yield hydrogen or hydroniumions; (d) has a
pH of less than 7.0; and (e) neutralizes bases or alkaline media.
Accuracy - Degree of conformity of a measure or test to a standard or true value.
Aliquot - A portion of a sample.
Alkali - Any compound having highly basic properties.
ATM - Atmosphere, pressure measurement.
Arsenic (As) - The major source of occupational exposure to arsenic is in the manufacture of
pesticides, herbicides, and other agricultural products.
Barium (Ba) - Barium is used in various alloys, in paints, soap, paper and rubber.
Base - A substance that usually liberates OH anions. Bases have a pH greater than 7.00.
BOD - Biochemical Oxygen Demand is a measure of the quantity of oxygen utilized in the
biochemical oxidation of organic matter related to the oxygen requirements in chemical
combustion, being determined entirely by the availability of the material as a biological food and
by the amount of oxygen utilized by the microorganisms during oxidation. BOD is the initial
quantity of oxygen used by polluted liquid immediately upon being introduced into water
containing dissolved oxygen. It may be exercised by end products of prior biochemical action or
chemical substances avid for oxygen. The BOD content is usually expressed in pounds per unit
of time, load of wastewater passing into a waste treatment system or to a body of water.
Biochemical Process - The process, by which, the metabolic activities of bacteria and other
microorganisms break down complex organic materials into simple, more stable substances.
Biodegradation - The destruction of mineralization of either natural or synthetic organic
materials by the microorganisms populating soils, natural bodies of water, or wastewater
treatment systems.
Carcinogen - A material that either causes cancer to humans or animals.
Catalyst - A substance that modifies (slows or accelerates) a chemical reaction without being
consumed.
Cadmium (Cd) - The main use of cadmium is in electroplating or galvanizing. It is also used as a
color pigment for paints and plastics and cathode material for nickel-cadmium batteries.
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COD - Chemical Oxygen Demand, a measure of the oxygen-consuming capacity of inorganic
and organic matter present in water or wastewater. It is expressed as the amount of oxygen
consumed from a chemical oxidant in a specific test. It does not differentiate between stable and
unstable organic matter and thus does not necessarily correlate with biochemical oxygen
demand. The method can be applied to domestic and industrial waste samples having an organic
carbon concentration greater than 15 mg/L. For lower concentrations of carbon such as in
surface water samples, the Low Level Modification should be used. Organic substances in the
sample are oxidized by Potassium Dichromate in 50% Sulfuric Acid solution at reflux
temperature. Silver Sulfate is used as a catalyst and Mercuric Sulfate is added to remove
Chloride interference. The excess Dichromate is titrated with standard Ferrous Ammonium
Sulfate, using Orthophenanthroline Ferrous complex as an indicator.
Chi Squared Test - In setting up methods you may want to compare sex or age variables on value
for a particular lab test. It is a statistical test most generally suitable for determining whether or
not an observed frequency of occurrence differs from that which is expected in accordance with
some hypothesis.
Chlorine (Cl) - An element ordinarily existing as a greenish-yellow gas about 2.5 times as heavy
as air. At atmospheric pressure and a temperature of - 30.1o F, the gas becomes an amber liquid
about 1.5 times as heavy as water.
Chlorine, available - A measure of the total oxidizing power of chlorinated lime and
hypochlorites.
Chlorination, breakpoint - Addition of chlorine to water or wastewater until the chlorine demand
has been satisfied and further additions result in a residual that is directly proportional to the
amount added beyond the breakpoint.
Chlorination, free residual - The application of chlorine or chlorine compounds to water or
wastewater to produce a free available chlorine residual directly or through the destruction of
ammonia or certain organic nitrogenous compounds.
Chlorine, residual - The amount of chlorine in all forms remaining in water after treatment to
ensure disinfection for a period of time.
Coefficient of Variation - An expression of standard deviation in terms as a percentage variance
from the mean value. The CV relates the Standard Deviation (SD) to the level at which the
measurements are made. CV = SD X 100
Composite - A sample made up of a collection of individual samples obtained at regular
intervals.
Composite (proportional) - A composite sample made up of sample whose volume is
proportional to the flow at the time of collection.
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Confidence Limits - Or confidence interval refers to the upper and lower values of the range
(interval) within which random variation are acceptable. Each lab and method may have its own.
The Town Branch Lab utilizes ± 2 Standard Deviation, which includes 95% of the values in a
Gaussian distribution.
Control - A fluid or substance whose physical and chemical properties closely resembles
unknown test specimen whose mean value has been assayed and is used as verification or check
on a test procedure.
Chromium (Cr) - Only the trivalent and hexavalent forms of chromium are of biologic
significance. Chromium in ambient air originates from industrial sources, are refining, and
combustion of fossil fuels.
Copper (Cu) - Wilson's disease is characterized by excessive accumulation of copper in liver,
brain, kidneys, and cornea.
Cyanide (CN-) - Cyanide is commonly found in certain rat and pest poisons, metal polishes,
photographic solutions and fumigating products.
Density - The ratio of weight (mass) to volume of a material.
DO - Dissolved Oxygen.
EPA – U.S. Environmental Protection Agency.
Iron (Fe) - Acute iron toxicity is nearly always due to accidental ingestion of iron-containing
medicines and most often occurs in children.
Filterable Residue - Dissolved solids and colloidal solids.
Fluoridation - The addition of a chemical to increase the concentration of fluoride ions in
drinking water to a pre-determined optimum limit to reduce the incidence of dental caries in
children.
Frequency distribution - A graphic (usually bar) representation of a set of values relating the
number of times each value is obtained to each value. Values are generally listed in ascending
order on X-axis and number of times the value occurs on Y-axis.
Gaussian distribution - Normal distribution curve, when data from biologic measurements are
plotted according to their frequencies, a bell-shaped curve is usually obtained. In all distributions
± 2 SD from the mean represents 95.45% of the population and ± 3 SD = 99.73%. (1 SD = 68%).
Grab - A single sample of wastewater taken at either a set time or flow.
Gravimetric - A means of measuring unknown concentration of a water quality indicator by
weighing.
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Halogenated Organics - A general term for organic molecules that contain one or more halogen
atoms.
Halogens - Chemical elements, either individually or collectively, composing group VIIB of the
periodic table. (i.e. Fluorine, Chlorine, Bromine, Iodine, and Astatine).
Hardness - A characteristic of water, imparted primarily by salts of calcium and magnesium,
such as bicarbonates, carbonates, sulfates, chlorides and nitrates, that causes curdling and
increased consumption of soap, deposition of scale in boilers, damage in some industrial
processes, and sometimes objectionable taste. It may be determined by a standard laboratory
titration procedure or computed from the amounts of calcium and magnesium expressed as
equivalent calcium carbonate.
Hepatotoxic - Causing liver damage.
LC50 - The concentration of a chemical in air or water that causes death to 50% of the animals.
Lead (Pb) - Lead is the most ubiquitous toxic metal. It is found everywhere, in food, the air, soil
and water. Sources include lead-based paint in old dwellings, combustion of lead-containing
auto exhausts or industrial emissions. Lead accumulates in bone and teeth. The target organs are
the kidneys, nervous and reproductive systems.
Mean - The average or sum of a group of observed values divided by the total number of
observations.
Median - The middle value of a series of numbers (arranged in ascending or descending order).
Note: If the distribution is not skewed.
Mercury (Hg) - The central nervous system is the major site of toxicity from exposure to
elemental mercury.
Milligrams per Liter - A unit of the concentration of water or wastewater constituents. It is 0.001
g of the constituent in 1,000 mL of water. It has replaced the unit formerly used commonly,
parts per million, to which it is approximately equivalent, in reporting the results of water and
wastewater analysis.
MSDS - Material Safety Data Sheet.
Mutagen - A material that induces genetic changes in the DNA of chromosomes.
Nickel (Ni) - Nickel is a respiratory tract carcinogen.
Nitrogen - An essential nutrient that is often present in wastewater as ammonia, nitrate, nitrite,
and organic nitrogen. The concentrations of each form and the sum, total nitrogen, are expressed
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as mg/L elemental nitrogen. Also present in some groundwater as nitrate and in some polluted
groundwater in other forms.
Nitrogen, ammonia - Quantity of elemental nitrogen present in the form of ammonia (NH3-).
Ammonia is a chemical combination of Hydrogen (H) and Nitrogen (N) occurring extensively in
nature. The combination used in water and wastewater engineering is expressed as NH3. The
protonated form (NH4)+, coexists with NH3 water and predominates under conditions of low pH.
Nitrogen Cycle - A graphical presentation of the conservation of matter in nature, form living
animal matter through dead organic matter, various stages of decomposition, plant life, and the
return of living animal matter, showing changes which occur in course of the cycle. It is used to
illustrate biological action and also aerobic and anaerobic acceleration of the transformation of
this element by wastewater and sludge treatment.
Nitrogen, Kjeldahl - A standard analytical method used to determine the concentration of the
organically-bound ammonia nitrogen state. The method covers the determination of total
Kjeldahl nitrogen in surface waters, domestic and industrial wastes, and saline waters. The
procedure converts nitrogen components of biological origin such as amino acids proteins and
peptides to ammonia, but may not convert the nitrogenous compounds of some industrial wastes
such as amines, nitro compounds, hydrazones, oximes, semi-carbazones and some refractory
tertiary amines. Total Kjeldahl is defined as the sum of free ammonia an organic ((NH4) 2S)4,
under the conditions of digestion.
Nitrogen, Organic - Nitrogen chemically bound in organic molecules such as proteins, amines,
and amino acids.
Nitrogen, Organic Kjeldahl - Defined as the difference obtained by subtracting the free ammonia
value (of Nitrogen, Ammonia) from the Total Kjeldahl Nitrogen value. This may be determined
directly by removal of ammonia before digestion.
Nitrogenous Oxygen - A quantitative measure of the amount of oxygen required for the Demand
(NOD) biological oxidation of nitrogenous material, such as ammonia nitrogen and organic
carbonaceous oxygen demand has been satisfied.
Non-Filterable Residue - Suspended solids.
Non-Volatile Residue - Fixed solids.
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Oil and Grease - In wastewater, a group of substances including fats, waxes, free fatty acids,
calcium and magnesium soaps, mineral oils and certain other non-fatty materials. The type of
solvent and method used for extraction should be stated for quantization. The method includes
the measurement of hexane extractable matter from waters, industrial wastes, and sewages. It is
applicable to the determination of relatively non-volatile hydrocarbons, animal fats and waxes,
grease and other types of greasy-oily matters. The method is not applicable to measurement of
light hydrocarbons that volatilize at temperatures below 80o C. The method covers the range
from 5 to 100 mg/L of extractable material.
Organic - Refers to volatile, combustible, and sometimes, biodegradable chemical compounds
containing carbon atoms (carbonaceous) bonded together and with other elements. The principal
groups of organic substances found in wastewater are proteins, carbon hydrates, and fats and
oils.
OSHA - Occupational Safety Health Administration.
ppb - Parts per billion.
ppm - Parts per million.
pH - A measure of the hydrogen-ion concentration in a solution, expressed as the logarithm (base
ten) of the reciprocal of the hydrogen-ion concentration in gram moles per liter. On the pH scale
(0 - 14), a value of 7 at 25o C represents a neutral condition. Decreasing values, below 7,
indicate increasing hydrogen-ion concentration (acidity); increasing values, above 7, indicate
decreasing hydrogen-ion concentration (alkalinity). Hydrogen-ion concentration is the weight of
hydrogen-ion in moles per liter of solution. Hydrogen-ion concentration is commonly expressed
as the pH value, which is the logarithm of the reciprocal of the hydrogen-ion concentration.
Phosphorus - An essential chemical element and nutrient for all life forms. Occurs in
orthophosphate, pyrophosphate, tripolyphosphate, and organic phosphate forms. Each of these
forms and their sum, total phosphorus, is expressed as mg/L elemental phosphorus. The method
covers the determination of specified forms of phosphorus in surface waters, domestic and
industrial wastes, and saline waters. They may be applicable to sediment-type samples, sludges,
algal blooms, etc., but sufficient data is not available at this time to warrant such usage when
measurements for phosphorus content are required. The methods are based on reactions that are
specific for the orthophosphate ion. Thus, depending on the prescribed pretreatment of the
sample, the various forms of phosphorus shown above may be determined. Except for in-depth
and detailed studies, the most commonly measured forms are phosphorus and dissolved
phosphorus, and orthophosphate and dissolved orthophosphate. Hydrolysable phosphorus is
normally found only in sewage-type samples and insoluble forms of phosphorus, as noted, are
determined by calculation. The methods are usable in the 0.01 to 0.5 mg/L P range.
Precision - Ability of self-duplication, amount of spread between replicates.
Range - Measure of dispersion of values and is merely the difference between the largest and the
smallest of a group of measurements.
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Reagent - Substance used in a chemical reaction to produce another substance or to detect its
composition.
Reliability - Measure of a method's ability to achieve both accuracy and precision.
Risk - The probability that a substance will produce harm.
Risk Assessment - Takes into account possible harmful effects on individuals or on society from
the use of a material in the quantity and in the manner proposed.
Safety - The reciprocal of risk, probability nothing will happen.
Selenium (Se) - Selenium derivatives are extremely Hepatotoxic.
Shifts - When 6 or more daily values of the same control distribute themselves on one side of the
mean value line, but are maintaining a constant level, indicates contamination of standard, use of
a new batch of reagents, changes in temperature of water bath, change in spectrophotometer
bulb, etc.
Solids - Material in a solid state. In water and wastewater treatment, any dissolved, suspended or
volatile substance contained in or removed from water or wastewater.
Solids, Colloidal - Finely divided solids intermediate between dissolved and suspended particles.
Solids, Dissolved - Solids that are present in solution.
Solids, Non-settleable - Wastewater matter that will stay in suspension for an extended period of
time. For laboratory purposes, 1 hour.
Solids, Settleable - Matter that will not stay in suspension for 1 hour. Matter that settles to the
bottom of an Imhoff Cone within 1 hour.
Solids, Suspended - Matter that is suspended in and will not settle in an hour. For laboratory
purposes it is that matter that can be collected on a standard filter.
Solids, Total - The sum of the dissolved and non-dissolved matter in wastewater. The sum of all
matter in a wastewater sample.
Solids, Volatile - The quantity of solids in wastewater lost on ignition of dry solids at 550o C.
This is a rough approximation of the organic matter.
Solids, Fixed - All matter remaining in a tare after ignition of dry solids at 550o C.
Suspended Solids - Insoluble solids that either float on the surface of, or are in suspension in
water, wastewater, or other liquids. Solid organic or inorganic particles (colloidal, dispersed,
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coagulated, flocculated) physically held in suspension by agitation or flow. The quantity of
material removed from wastewater in a laboratory test, expressed as milligrams per liter and
referred to as nonfilterable residue. This method is applicable to surface waters, domestic
industrial wastes and saline waters. The practical range of the determination is 20 mg/L to
20,000 mg/L. A well-mixed sample is filtered through a standard glass fiber filter, and the
residue retained on the filter is dried to a constant weight at 103 to 105o C. Non-homogenous
particulates such as leaves, sticks, fish, and lumps of fecal matter should be excluded from the
sample. Preservation of the sample is 4o C for a maximum of 7 days.
Standard - A highly purified material, having a known, weighed, fixed value or concentration of
the substance being analyzed.
Standard Deviation - A measure of the scatter of individual values in a Gaussian distribution, the
square root of the arithmetic mean of the square of the deviation from the arithmetic mean
divided by n-1.
Standard Deviation Interval - Measure of difference between our lab mean and the average of all
lab means in a comparison group. The difference is expressed in terms of the SD of all the means
in the group.
Tare - A clean, dry container used in a gravimetric analysis.
T-test - Used to ascertain whether the means of two sample groups differ significantly.
Calculated using the mean and SD of two matched groups. Refer to Tietz.
TI - Therapeutic Index is the ratio of the dose required to produce a toxic effect.
Total Oxygen Demand (TOD) - a quantitative measure of all oxidizable material in a sample of
water or wastewater as determined instrumentally by measuring the depletion of oxygen after
high-temperature combustion, as TOC, COD.
Total Carbon (TC) - a quantitative measure (mg/L) of both total inorganic (TIC) and total
organic carbon (TOC) in water or wastewater. Determined instrumentally by chemical oxidation
to CO2 and subsequent infrared detection in a carbon analyzer.
Total Organic Carbon - The amount of carbon bound in organic compounds in a sample. Because
all organic compounds have carbon as the common element, total organic carbon measurements
provide a fundamental means of accessing the degree of organic pollution. The carbonaceous
analyzer measures all of the carbon in a sample after injection into the combustion tube.
Because of various properties of carbon-containing compounds in liquid samples, preliminary
treatment of the sample prior to injection dictates the definition of the carbon as it is measured.
The final usefulness of the carbon measurement is in assessing the potential oxygen-demanding
load of organic material on a receiving stream. This statement applies whether the carbon
measurement is made on a sewage plant effluent, industrial waste, or on water taken directly
from the stream. In this light, carbonate and bicarbonate carbon are not a part of the oxygen
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demand in the stream and therefore should be discounted in the final calculation or removed
prior to analysis. The manner of preliminary treatment therefore defines the types of carbon
which are measured.
Toxicant - A substance that kills or injures an organism through chemical or physical or
biological action. Examples include cyanides, pesticides, and heavy metals (such as zinc and
chromium).
Toxicology - Is the basic science of poisons and their effects. It is the study of the physical
effects of chemicals on biological systems.
Trace Element - Any element in water or wastewater that for reasons associated with natural
distribution, industrial use, solubility, or other factors, is present at very low concentrations as an
essential element.
Trends - When charting quality control values for the same control, the values of the control
tended to increase or decrease over a period of 6 consecutive days - can indicate deterioration,
changes in stability of standard, incomplete protein precipitation, etc.
Zinc (Zn) - Zinc is ubiquitous in the environment so that it is present in most foodstuffs, water
and air.
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Section 23
Bibliography
Code of Federal Regulations (CFR). 2003. Guidelines Establishing Test Procedures for the
Analysis of Pollutants. 40 CFR 136.3, Title 40, Chapter 1. U.S. Environmental Protection
Agency. pg 5-337.
Fisher Scientific. AR50 Fisher Scientific User Manual. Fisher Scientific, Inc.
HACH AutoCat 9000 Chlorine Amperometric Titrator Instruction Manual. HACH Company,
Loveland, CO.
HACH DR/4000 Handbook, Method 8083, Procedure Code N.3, Nessler Method following
distillation. HACH Company, Loveland, CO.
HACH DR/4000 procedure method 8027. HACH Company, Loveland, CO.
HACH DR/4000 Procedure Manual, EPA approved Method 8190 Phosphorus, Total. HACH
Company, Loveland, CO.
HACH HQ Series Portable Meter Users Manual, September 2006, Edition 5. HACH Company,
Loveland, CO.
HACH Water Analysis Handbook. Method 8023, Chromium Hexavalent (1,5Diphenylcarbohydrazide Method). 2nd Edition. 1992. HACH Company, Loveland, CO.
HACH Water Analysis Handbook. Method 8213 Hardness, Total with a Digital Titrator. 2nd
Edition. 1992. HACH Company, Loveland, CO.
HACH Water Analysis Handbook. Method 8291, Volatile Acids, Buret Titration Method. 2nd
Edition. 1992. HACH Company, Loveland, CO.
Memorandum: EPA Recommendation for the use of HACH method 10360 [Revision 1.1,
January 2006] (ATP Case # N04-0013).
Report on the Validation of Proposed EPA Method 360.3 (Luminescence) for the Measurement
of Dissolved Oxygen in Water and Wastewater. August 2004. HACH Company, Loveland, CO.
Lachat Micro Dist User Manual, Method Cyanide in Waters (MICRO DIST Cyanide-1). Lachat
Instruments, HACH Company, Loveland, CO.
Orion Research Incorporated Laboratory Products Group.
Standard Methods 2320-B. APHA-American Public Health Association Standard Methods for
the Examination of Water and Wastewater; 21th edition ed.; American Water Works Association
and Water Pollution Control Federation: Washington, DC, 2005.
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Standard Methods 2340-C. APHA-American Public Health Association Standard Methods for
the Examination of Water and Wastewater; 21th edition ed.; American Water Works Association
and Water Pollution Control Federation: Washington, DC, 2005.
Standard Methods Part 2540-B. Total Solids dried at 103 oC to 105 oC. APHA-American Public
Health Association Standard Methods for the Examination of Water and Wastewater; 21th
edition ed.; American Water Works Association and Water Pollution Control Federation:
Washington, DC, 2005.
Standard Methods Part 2540-D. Total Suspended Solids Dried at 103 oC to 105 oC. APHAAmerican Public Health Association Standard Methods for the Examination of Water and
Wastewater; 21th edition ed.; American Water Works Association and Water Pollution Control
Federation: Washington, DC, 2005.
Standard Methods Part 2540-E, Fixed and Volatile Solids Ignited at 550 oC. APHA-American
Public Health Association Standard Methods for the Examination of Water and Wastewater; 21th
edition ed.; American Water Works Association and Water Pollution Control Federation:
Washington, DC, 2005.
Standard Methods Part 2540-F. Settleable Solids. APHA-American Public Health Association
Standard Methods for the Examination of Water and Wastewater; 21th edition ed.; American
Water Works Association and Water Pollution Control Federation: Washington, DC, 2005.
Standard Methods Part 3500 B. Chromium, Colorimetric Method. APHA-American Public
Health Association Standard Methods for the Examination of Water and Wastewater; 21th
edition ed.; American Water Works Association and Water Pollution Control Federation:
Washington, DC, 2005.
Standard Methods 4500-Cl D. Chlorine Residual Amperometric Titration Method. APHAAmerican Public Health Association Standard Methods for the Examination of Water and
Wastewater; 21th edition ed.; American Water Works Association and Water Pollution Control
Federation: Washington, DC, 2005.
Standard Methods Part 4500-P. Phosphorous. APHA-American Public Health Association
Standard Methods for the Examination of Water and Wastewater; 21th edition ed.; American
Water Works Association and Water Pollution Control Federation: Washington, DC, 2005.
Standard Methods 5210-A. and 5210-B. Biochemical Oxygen Demand (BOD). APHA-American
Public Health Association Standard Methods for the Examination of Water and Wastewater; 21th
edition ed.; American Water Works Association and Water Pollution Control Federation:
Washington, DC, 2005.
Standard Methods Part 5560-C, Distillation Method. APHA-American Public Health Association
Standard Methods for the Examination of Water and Wastewater; 21th edition ed.; American
Water Works Association and Water Pollution Control Federation: Washington, DC, 2005.
52
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U.S. EPA. Analysis of Water and Wastes. EPA-600-4-79-020. U.S. Environmental Protection
Agency; Office of Research and Development, Washington, DC, 1982.
U.S. EPA. Method 130.2. Methods for Chemical Analysis of Water and Wastes. EPA-600-4-79020. U.S. Environmental Protection Agency; Office of Research and Development, Washington,
DC, 1982.
U.S. EPA. Method 150.1 pH (Electrometric) Issued 1971 (Editorial revision 1978 and 1982).
Methods for Chemical Analysis of Water and Wastes. EPA-600-4-79-020; U.S. Environmental
Protection Agency; Office of Research and Development, Washington, DC, 1982.
U.S. EPA Method 160.2 Residue, Non-Filterable & Total Suspended Solids. Issued 1971.
Methods for Chemical Analysis of Water and Wastes. EPA-600-4-79-020. U.S. Environmental
Protection Agency; Office of Research and Development, Washington, DC, 1982.
U.S. EPA Method 160.3 Residue, Total (Gravimetric, Dried at 103 – 105 oC). Issued 1971.
Methods for Chemical Analysis of Water and Wastes. EPA-600-4-79-020. U.S. Environmental
Protection Agency; Office of Research and Development, Washington, DC, 1982.
U.S. EPA Method 160.4 Residue, Volatile, (Gravimetric, Ignition at 550 oC), Issued 1971.
Methods for Chemical Analysis of Water and Wastes. EPA-600-4-79-020. U.S. Environmental
Protection Agency; Office of Research and Development, Washington, DC, 1982.
U.S. EPA. Method 310.1. Methods for Chemical Analysis of Water and Wastes. EPA-600-4-79020. U.S. Environmental Protection Agency; Office of Research and Development, Washington,
DC, 1982.
U.S. EPA Method 330.1 Chlorine, Total Residual (Titrimetric, Amperometric) Issued 1978.
Methods for Chemical Analysis of Water and Wastes. EPA-600-4-79-020. U.S. Environmental
Protection Agency; Office of Research and Development, Washington, DC, 1982.
U.S. EPA. Method 335.4. Revision 1.0, August 1993. Methods for Chemical Analysis of Water
and Wastes. EPA-600-4-79-020. U.S. Environmental Protection Agency; Office of Research and
Development, Washington, DC, 1982.
U.S. EPA Method 350.2. Distillation procedure. Methods for Chemical Analysis of Water and
Wastes. EPA-600-4-79-020. U.S. Environmental Protection Agency; Office of Research and
Development, Washington, DC, 1982.
U.S. EPA Method 365.2 Phosphorous, All Forms (Colorimetric, Ascorbic Acid, Single Reagent)
Revised March 1983. Methods for Chemical Analysis of Water and Wastes. EPA-600-4-79-020.
U.S. Environmental Protection Agency; Office of Research and Development, Washington, DC,
1982.
U.S. EPA Method 405.1, Biochemical Oxygen Demand, 5 Days @ 20ºC, Issued 1971, Editorial
revision 1974. Methods for Chemical Analysis of Water and Wastes. EPA-600-4-79-020. U.S.
53
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Environmental Protection Agency; Office of Research and Development, Washington, DC,
1982.
U.S. EPA. Microbiological Methods for Monitoring the Environment: Water and Wastes. EPA
600/8-78-017. U.S. Environmental Protection Agency; Environmental Monitoring and Support
Laboratory, Office of Research and Development, Washington, DC, 1978. page 124.
Water Pollution Control Federation. Simplified Laboratory Procedures for Wastewater
Examination, Third Edition. 1985.
54
APPENDIX A
LABORATORY ANALYSIS LIST
A-1
TB Laboratory QAP - Appendix A
Revision 1
September 16, 2009
Table A1. Town Branch Laboratory Analysis List
Sample Name
Analysis
Analysis Requirements
Raw Influent
CBOD5
TSS
Ammonia
Required
Required
Required
Plant Effluent
pH
Dissolved Oxygen
CBOD5
TSS
Ammonia
Total Phosphorus
Hardness
Fecal Coliforms
Residual Chlorine
Metals - Dissolved
Metals - Total
Required
Required
Required
Required
Required
Required
Required
Required
Required
Required
Required
Raw Influent
pH
Dissolved Oxygen
Settleable
Total Phosphorus
Ortho-phosphate
Metals - Dissolved
Metals - Total
Operation
Operation
Operation
Operation
Operation
Operation
Operation
Primary Influent
CBOD5
Settleable
TSS
Operation
Operation
Operation
Primary Effluent
CBOD5
Settleable
TSS
Operation
Operation
Operation
Plant Effluent
Settleable
Ortho-phosphate
Operation
Operation
A-2
TB Laboratory QAP - Appendix A
Revision 1
Sample Name
September 16, 2009
Analysis
Analysis Requirements
Creek Above Plant
pH
Dissolved Oxygen
CBOD5
TSS
Ammonia
Settleable
Total Phosphorus
Ortho-phosphate
Hardness
Fecal Coliforms
Metals - Total
Operation
Operation
Operation
Operation
Operation
Operation
Operation
Operation
Operation
Operation
Operation
Tap Water
Total Phosphorus
Operation
Mixed Liquor
pH
Total Alkalinity-Inf.
Total Alkalinity-Eff.
Settleable
MLTSS
MLVSS
SVI
SDI
Rise Time
Micro Exam
Operation
Operation
Operation
Operation
Operation
Operation
Operation
Operation
Operation
Operation
Return Activated Sludge
(R.A.S.)
pH
TSS
VSS
% Total Solids
Operation
Operation
Operation
Operation
Raw Sludge to Thickener
pH
% Total Solids
% Volatile Solids
Operation
Operation
Operation
Raw Sludge Thicken Sludge
% Total Solids
Operation
Raw Sludge
Thickener Overflow
TSS
Total Phosphorus
Operation
Operation
Combined Sludge Density Meter
% Total Solids
% Volatile Solids
Operation
Operation
A-3
TB Laboratory QAP - Appendix A
Revision 1
Sample Name
September 16, 2009
Analysis
Analysis Requirements
#1 Primary Digester
pH
Volatile Acids
Total Alkalinity
VA/ALK Ratio
% Total Solids
% Volatile Solids
Operation
Operation
Operation
Operation
Operation
Operation
#2 Primary Digester
pH
Volatile Acids
Total Alkalinity
VA/ALK Ratio
% Total Solids
% Volatile Solids
Operation
Operation
Operation
Operation
Operation
Operation
#3 Primary Digester
pH
Volatile Acids
Total Alkalinity
VA/ALK Ratio
% Total Solids
% Volatile Solids
Operation
Operation
Operation
Operation
Operation
Operation
Digested Sludge
pH
% Total Solids
% Volatile Solids
% Removal of Volatile Solids
Operation
Operation
Operation
Operation
% Total Solids
Operation
TSS
Total Phosphorus
Operation
Operation
Filter Belt Press
Belt #1 Cake Solids
% Total Solids
Operation
Belt #2 Cake Solids
% Total Solids
Operation
Belt #3 Cake Solids
% Total Solids
Operation
Belt #4 Cake Solids
% Total Solids
Operation
Filter Belt Press Cake
Metals - Total
Operation
Belt Feed and Wastewater
Belt Feed Solids
Filtrate
A-4
TB Laboratory QAP - Appendix A
Revision 1
September 16, 2009
Table A2. West Hickman WWTP Analysis List
Sample Name
Analysis
Analysis Requirements
Raw Influent
CBOD5
TSS
Required
Required
Plant Effluent
CBOD5
TSS
Total Phosphorus
Ammonia
Hardness
Fecal Coliforms
Metals - Dissolved
Metals - Total
Required
Required
Required
Required
Required
Required
Required
Required
Raw Influent
Ammonia
Settleable
Total Phosphorus
Ortho-phosphate
Volatile Acids
Total Alkalinity
Metals - Dissolved
Metals - Total
Operation
Operation
Operation
Operation
Operation
Operation
Operation
Operation
Combined Influent
CBOD5
TSS
Operation
Operation
CBOD5
Total Phosphorus
Ortho-phosphate
Volatile Acids
Total Alkalinity
Operation
Operation
Operation
Operation
Operation
CBOD5
Total Phosphorus
Ortho-phosphate
Volatile Acids
Total Alkalinity
Operation
Operation
Operation
Operation
Operation
Biological Phosphorus
Removal - Influent
Biological Phosphorus
Removal - Effluent
A-5
TB Laboratory QAP - Appendix A
Revision 1
Sample Name
September 16, 2009
Analysis
Analysis Requirements
TSS
VSS
Total Alkalinity
Operation
Operation
Operation
Operation
Zone #2 Effluent
MLTSS
(Formerly Zone #2 Mixed Liquor) MLVSS
Total Alkalinity
Operation
Operation
Operation
Plant Effluent
Settleable
Ortho-phosphate
Operation
Operation
Creek Above Plant
CBOD5
TSS
Metals - Total
Operation
Operation
Operation
Creek Below Plant
CBOD5
TSS
Metals - Total
Operation
Operation
Operation
Return Activated Sludge
(R.A.S.)
pH
Total Alkalinity
TSS
VSS
% Total Solids
% Volatile Solids
Operation
Operation
Operation
Operation
Operation
Operation
Belt Feed and Wastewater
Combined Sludge Feed
% Total Solids
% Volatile Solids
Operation
Operation
Filtrate
TSS
Total Phosphorus
Ortho-phosphate
Operation
Operation
Operation
Filter Belt Press
Belt #1 Cake Solids
% Total Solids
Operation
Belt #2 Cake Solids
% Total Solids
Operation
Belt #3 Cake Solids
% Total Solids
Operation
Filter Belt Cake
Metals - Total
Operation
Zone #1 Effluent
(Formerly Zone #1 M/L)
A-6
TB Laboratory QAP - Appendix A
Revision 1
September 16, 2009
Table A3. Blue Sky WWTP Analysis List
Sample Name
Analysis
Analysis Requirements
Raw Influent
CBOD5
TSS
Ammonia
Required
Required
Required
Plant Effluent
CBOD5
TSS
Ammonia
Hardness
Fecal Coliforms
Oil & Grease
Total Phenols
Total Phosphorus
Ammonia
Metals - Dissolved
Metals - Total
Required
Required
Required
Required
Required
Required
Required
Required
Required
Required
Required
Raw Influent
Total Alkalinity
Total Phosphorus
Ortho-phosphate
Operation
Operation
Operation
Plant Effluent
Ortho-phosphate
Operation
Contact Zone
TSS
Total Alkalinity
Operation
Operation
Re-Aeration
TSS
Total Alkalinity
Operation
Operation
Digester
TSS
Total Alkalinity
Operation
Operation
A-7
TB Laboratory QAP - Appendix A
Revision 1
September 16, 2009
Table A4. Data Provided to the Laboratory by Operations Staff
Town Branch WWTP
Total Influent Flow
Peak Flow
Rainfall
West Hickman WWTP
Total Influent Flow
Peak Flow
Rainfall
Nitrification Return Flow
Nitrification Waste Flow
Total Pounds Chlorine
Total Pounds SO2
Number of Tanks in Service
Raw Influent - pH
Raw Influent - DO
Combined Influent - pH
Combined Influent - DO
Zone #1 Effluent - 30 min. Settleable
Zone #2 Effluent - pH
Zone #2 Effluent - 30 min. Settleable
Zone #2 Effluent - Rise Time
Plant Effluent - pH
Plant Effluent - DO
Plant Effluent - TP
Plant Effluent - Fecal Coliforms
Chlorine Contact - Residual Chlorine
Plant Effluent - Residual Chlorine
Creek Above Plant - pH
Creek Above Plant - DO
Creek Below Plant - pH
Creek Below Plant – DO
Blue Sky WWTP
Total Influent Flow
Rainfall
Raw Influent - pH
Raw Influent - DO
Plant Effluent - pH
Plant Effluent - DO
Plant Effluent - Residual Chlorine
A-8
TB Laboratory QAP - Appendix C
Revision 1
APPENDIX B
September 16, 2009
STAFF RESPONSIBILITIES
B-1
TB Laboratory QAP - Appendix C
Revision 1
September 16, 2009
Appendix B. Staff Responsibilities
Class Title: LABORATORY SUPERVISOR
Reports To: Division of Water Quality Director
Supervision Exercised: Laboratory Technicians
General Function:
Manage the operation and maintenance of the government's wastewater laboratory.
Performs work of moderate to considerable difficulty in conducting and supervising
Laboratory Technicians in a variety of chemical/bacteriological tests and analyses
on water released from wastewater treatment plants to determine conformance to
EPA mandated standards. Also conducts and supervises Laboratory Technicians in
the monitoring, testing and analysis of industrial waste and other discharges into
the sanitary sewer systems to determine compliance with local, State and EPA
permits and other applicable rules/regulations.
Essential Functions:
Administration/Supervision:
• Set up a laboratory budget and maintain budget records
• Set up and maintain laboratory personnel records - Do employee evaluations. - Deal with
employee problems. - Do job interviews for new employees. - Set up employee work
schedules.
• Set up and maintain all necessary records and reports as required by the state and EPA.
• Prepares and submits all required records and reports to state and Environmental
Protection Agency.
• Do all ordering of supplies and specification of equipment for the laboratory and also for
other departments wanting lab related equipment
• Supervises and trains Laboratory Technicians on laboratory and testing procedures
• Supervises the cleaning and sterilization of laboratory equipment, utensils and facilities
• Supervises the storage of all chemicals insuring that all required safety and storage
procedures are followed
• Supervises the proper collection of sewage and sludge samples during various phases of
wastewater treatment
Program Coordination:
• Manage the LFUCGs laboratories for testing the quality of waters released from three
different wastewater treatment plants.
• Maintain and evaluate data on these plants.
• Create and modify programs to sort, store, and calculate data, monitor trends, and
generate graphs on personal computer.
• Help other departments set up programs to use data provided by the laboratory. Work
with operations staff of three plants and our engineers to help them use the data provided
by the laboratory.
• Make suggestions as to how the data could be used to deal with operational problems.
B-1
TB Laboratory QAP - Appendix C
Revision 1
September 16, 2009
Sample Preparation, Testing and Analysis:
• Check and evaluate all lab data. Run standard to make calibration curve to establish
quality of the test.
• Train technicians to run analyses and supervise the procedures.
• Determine how samples are to be prepared for analysis.
• Supervise the sample preparation.
• Determine all laboratory analysis procedures to be used in the analysis of samples. Train
the lab personnel on how to use these procedures and supervise their use.
• Set up and run sophisticated bacteriology, chemical, and microbiology analyses that can
only be run by the laboratory supervisor; record and report the data.
Miscellaneous:
• Travels to West Hickman WWTP Laboratory to calibrate equipment and/or run analyses
on request.
• Maintains special laboratory programs such as safety, quality control or quality
assurance.
• Answers telephone inquiries and relays messages and information.
• Responsible for knowing and complying with all UCG and division safety rules and for
attending safety meetings. Performs other related duties as required.
Knowledge, Skills and Abilities:
• Considerable knowledge of the principles, practices and techniques of bacteriology and
chemistry; the operation and maintenance of complex chemical laboratory equipment;
and the safety requirements of a chemical laboratory in the performance of various tests
and the storage and use of a variety of chemicals
• Fundamental knowledge of the design and operation of an AA or ICP to facilitate repair
and/or assistance in troubleshooting
• Knowledge in the use of micro or personal computers to sort data, monitor trends,
generate reports, modify existing and create new programs using spreadsheet applications
• Good knowledge of State and Environmental Protection Agency (EPA) quality control
procedures, record keeping and report requirements
• Good observational skills to detect problems in analytical procedures and to detect
possible errors in data entry, sample processing, etc.
• Ability to establish and monitor new testing procedures to meet state and EPA
requirements
• Ability to use a variety of complex laboratory equipment and understand, set up and run
all standard laboratory tests on wastewater, sludge and industrial waste as required by
regulatory agencies
• Ability to train and supervise Laboratory Technicians
• Ability to establish and maintain effective working relationships with regulatory
agencies, suppliers, industries that discharge waste into the sanitary sewer system and
fellow employees.
B-2
TB Laboratory QAP - Appendix C
Revision 1
Communications:
Who (Title)
Communicate About What
Director
Reports/data
Deputy Director
Reports/date/special requests
Municipal Engineer
Reports/data/computers
Pretreatment Manager
Data/special requests
Plant Operations Supervisors
Data/operations
Sludge Disposal Supervisors
Data/operations
Engineering Technicians
Special requests
Administrative Office Assistants
General/office work
Vendors/Suppliers
Purchasing / pricing / discount
Labs from other cities
Operation Problems
State Inspectors
Operations / paperwork
EPA Inspectors
Operations / paperwork
Commercial Labs
Data on an industry
September 16, 2009
How Often
Occasionally
Weekly to daily
Daily
Daily to weekly
Daily
Daily
Weekly
Daily
Weekly
Occasionally
Monthly
Occasionally
Monthly
Special Requirements:
•
•
•
•
•
•
•
May be required to possess a valid driver's license.
Physical strength and agility to allow considerable walking, lifting, climbing and working
with samples that are odorous and hazardous.
Must be able to work weekends and holidays and perform on-call duty.
Must be able to operate Urban County Government equipment and vehicles in a safe,
prudent and responsible manner.
All positions require drug testing before employment and will require a preemployment physical as stated in Ordinances 2114(b) and 23-16.
Pursuant to the Drug Free Workplace Act of 1988 and to sections 21-52, 22-34 and
23-50 of the Code of Ordinances, all employees must remain drug and alcohol free
when reporting to work, while at work and while engaged in any work related activities.
Based on Federal Regulation 19-10, some positions in this classification are eligible
for and will be offered the hepatitis vaccinations. In addition, employees will be
required to sign a statement stating they have accepted or declined the hepatitis
vaccination.
B-3
TB Laboratory QAP - Appendix C
Revision 1
September 16, 2009
Class Title: LABORATORY TECHNICIAN
Reports To: Laboratory Supervisor, Plant Operations Supervisor or other supervisor
Supervision Exercised: None
General Function:
Performs technical work of considerable difficulty in conducting a variety of chemical,
bacteriological, and industrial waste analyses for wastewater treatment plants to assure
compliance with local, State, and Environmental Protection Agency permits.
Essential Functions:
Sample Gathering / Preparation:
• Collects samples and makes sure that samples collected by others are in
accordance with government regulations.
• Prepares wastewater, sludge, and pretreatment samples for analyses.
• Cleans laboratory equipment and work area.
• Sterilizes bacteriological equipment.
• Calibrates laboratory equipment.
• Prepares reagents and mixtures as required for laboratory testing.
Sample Testing/Analysis:
• Conducts a variety of laboratory analyses in accordance with government regulations
to include pH, biochemical, oxygen demand, suspended and total solids, volatile solids,
chemical concentrations, alkalinity and other.
• Conducts routine bacteriological
examination on wastewater samples.
Conducts microorganism counts of activated
sludge.
• Makes dissolved oxygen and pH readings in the field when requested.
Recording/Reporting Results:
• Records all data on worksheets and monthly sheets.
• Prepares monthly reports.
• Maintains inventory of supplies by informing Laboratory Supervisor when inventories
get low.
Miscellaneous:
• Travels to West Hickman WWTP Laboratory to calibrate equipment and/or run analyses
on request.
• Maintains special laboratory programs such as safety, quality control or quality
assurance.
• Answers telephone inquiries and relays messages and information.
• May be assigned to perform duties of higher or lower level classifications in this or
related class series and/or perform duties on a temporary or acting basis in accordance
B-4
TB Laboratory QAP - Appendix C
Revision 1
•
September 16, 2009
with Ordinances 21-15 and 21-16 and/or act as a lead worker.
Responsible for knowing and complying with all UCG and division safety rules and for
attending safety meetings. Performs other related duties as required.
Physical Demands and Working Conditions:
•
•
•
Physical demands include standing, lifting, carrying, and walking.
Must have the strength and agility to allow the employee to perform job duties which
would be found at the level of medium work (i.e. exerting 20 to 50 pounds of force) as
defined in the PAQ.
Working conditions include performing a majority of job duties indoors with wet and
humid surroundings, fumes, odors, and chemicals.
Knowledge, Skills and Abilities:
Considerable knowledge of:
• the occupational hazards and safety precautions necessary relative to the area of
assignment;
• the elementary principles, practices and techniques of bacteriology and chemistry;
• the operation, maintenance and calibration of standard and complex chemical laboratory
equipment.
Good knowledge of:
• the elementary principles, practices and techniques of bacteriology and chemistry;
• personal computers and related software used in the field;
• the operation and care of standard chemical laboratory equipment.
Ability to:
• perform standard chemical and bacteriological tests accurately;
• follow oral and written instructions exactly;
• prepare reports on the results of laboratory tests performed;
• establish and maintain effective working relationships with fellow employees and other
involved parties;
• understand, set up and run standard laboratory tests on wastewater and sludge per
"standard methods".
Special Requirements:
•
•
•
•
•
May be required to possess a valid driver's license.
Physical strength and agility to allow considerable walking, lifting, climbing and working
with samples that are odorous and hazardous.
Must be able to work weekends and holidays and perform on-call duty.
Must be able to operate Urban County Government equipment and vehicles in a safe,
prudent and responsible manner.
All positions require drug testing before employment and will require a pre-
B-5
TB Laboratory QAP - Appendix C
Revision 1
•
•
September 16, 2009
employment physical as stated in Ordinances 2114(b) and 23-16.
Pursuant to the Drug Free Workplace Act of 1988 and to sections 21-52, 22-34 and
23-50 of the Code of Ordinances, all employees must remain drug and alcohol free
when reporting to work, while at work and while engaged in any work related activities.
Based on Federal Regulation 19-10, some positions in this classification are eligible
for and will be offered the hepatitis vaccinations. In addition, employees will be
required to sign a statement stating they have accepted or declined the hepatitis
vaccination.
B-6
TB Laboratory QAP - Appendix C
Revision 1
September 16, 2009
APPENDIX C CURRENT PERSONNEL AND
LAB APPROVED SIGNATURES
C-1
TB Laboratory QAP - Appendix C
Revision 1
September 16, 2009
Appendix C. Current Personnel and Laboratory's Approved Signatures
Job Title: Laboratory Supervisor
Name: Dr. David J. Price
Employee #: 46274
Education: Ph.D. Biology, University of Kentucky, 2008
Signature:
Job Title: Laboratory Technician - QA/QC Manager
Name: La Vada M. Green
Employee #: 22849
Education: B.S. Physics, Eastern Kentucky University,
Signature:
Job Title: Laboratory Technician – Safety Officer
Name: Di-Linh Cao-Nguyen
Employee #: 39494
Education: B.S. Biology, University of Kentucky, 1997
Signature:
C-2
TB Laboratory QAP - Appendix C
Revision 1
September 16, 2009
Job Title: Laboratory Technician - Microbiology
Name: Maria Lundin
Employee #: 45854
Education: B.S. Biology, University of Wisconsin-Green Bay, 1995
Signature:
Job Title: Laboratory Technician
Name: Jerry W. McDaniel
Employee #: 35894
Education: Navy training, Medical Technology, 1974
Signature:
Job Title: Laboratory Technician – Database management
Name: Brian Reynolds
Employee #: 43385
Education: B.S. Biology, Eastern Kentucky University, 2006
Signature:
C-3
TOWN BRANCH LABORATORY STANDARD
OPERATING PROCEDURES
1
SOP – Alkalinity
LFUCG Laboratory
Page 2 of 219
Revision Number 2
Last Revised 9/09
Alkalinity (Titrimetric)
Standard Methods 2320-B
1.
Scope, Significance to Process and Application
1.1
2.
3.
Summary of Method
2.1
Executive Summary
The principle of operation for the Orion Test Kit is the same as the conventional
titration. A pre-measured volume of reagent is added to the sample. This reagent
is composed of several acids that react with the alkaline species in the sample,
resulting in a change in sample pH. The observed pH reading after the addition of
the reagent varies directly with the total alkalinity. Each pH reading corresponds
to a unique value for alkalinity, expressed in mg/L (ppm) CaCO3. The alkalinity
values are obtained in a chart that cross references with the pH values.
2.2
Discussion
Alkalinity of water is its acid-neutralizing capacity. Raw domestic wastewater has
an alkalinity less than or slightly higher than in the water supply. Properly
operating anaerobic digesters have a supernatant alkalinity in the range of 20004000 mg CaCO3/L. Some samples are diluted. No color change is noted.
Health & Safety Precautions
3.1
3.2
4.
Watch out for broken glass from beakers and cylinders.
Wastewater samples have the potential to be hazardous, use appropriate caution.
Sample Handling and Preservation
4.1
5.
This method is applicable to drinking, surface, and saline waters, as well as,
domestic and industrial wastes.
Samples should be run as soon as possible.
Reagents
5.1
5.2
5.3
Total Alkalinity Reagent (Orion # 700011)
Alkalinity Standard/Control (1000 ppm; Orion # 700012)
Nanopure Water
2
SOP – Alkalinity
LFUCG Laboratory
Page 3 of 219
Revision Number 2
Last Revised 9/09
6.
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
7.
Interferences
7.1
7.2
8.
Fisher AR50 ph Meter using a glass electrode and reads to 0.05 pH units
100 mL volumetric flask (for standard)
100 mL graduated cylinders
Appropriate size beakers to contain samples and reagents
Repipettors (1 mL, 5 mL, and 10 mL) and tips
Substances, such as salts of weak organic and inorganic acids present in large
amounts, may cause interference in the electrometric pH measurements.
Oil and Grease, by coating the pH electrode may also interfere, causing a sluggish
response.
Procedures
8.1
Steps
1) Calibrate pH meter (see SOP-pH, Section 8).
2)
Check alkalinity of Nanopure water. (Any alkalinity in the water used for
diluting will contribute to the total alkalinity measured in the control)
a.
b.
c.
d.
e.
3)
Measure 100 mL of Nanopure water into 150 mL beaker.
Add 1.0 mL Alkalinity Reagent and insert pH probe.
Turn on stirrer.
Read pH value when the meter displays “STABLE”.
Determine total alkalinity of the Nanopure water using the Low-level
chart (0-25 ppm), and multiply result by 0.9. This value (AH2O) is the
contribution to total alkalinity from Nanopure water.
Check alkalinity of Standard.
a. Measure 10 mL of Alkalinity Standard into 100 mL volumetric flask
and dilute to mark with Nanopure water.
b. Pour standard into 150 mL beaker.
c. Add 10 mL of Total Alkalinity Reagent and insert pH probe.
d. Turn on stirrer.
e. Read pH value when the meter displays “STABLE”.
f. Obtain alkalinity value (A) from the Full chart (0-225 ppm) by cross
referencing with pH value.
g. Determine alkalinity of standard (Astd):
Astd = A - AH2O (Value should be 100 ± 5 ppm).
3
SOP – Alkalinity
LFUCG Laboratory
Page 4 of 219
Revision Number 2
Last Revised 9/09
4)
Record alkalinity value, date, time and initials in bench sheet.
5)
For samples:
a. Determine sample volume from the bench sheet (generally, 100 mL).
b. To make a 50:1 dilution: pipette 2 mL of sample and dilute to 100 mL
volume with Nanopure water.
c. Pour sample into 150 mL beaker.
d. Add 10 mL of Total Alkalinity Reagent and insert pH probe.
e. Turn on stirrer.
f. Read pH value when the meter displays “STABLE”.
g. Obtain alkalinity value (Asmp) from the Full chart (0-225 ppm) by
cross referencing with pH value.
h. If diluted, determine alkalinity of diluted sample (Ads):
Ads = Asmp - AH2O
6)
8.2
9.
Record alkalinity value, date, time and initials in bench sheet.
Helpful Hints
1) Alkalinity ranges for the standard should be 100 ± 5 ppm, if not, try
recalibrating and use clean glassware.
QA/QC Requirements
Analysis values for the standard must have a pH range of 4.41 ± 0.05 (equivalent to 930 1070 mg/L CaCO3). If this criterion is not met, corrective action is indicated. See Quality
Assurance Program (QAP) Sec. 15 “Corrective Action Policies and Procedures”.
10.
Expected Results
10.1
KPDES Permit Requirements
None required.
10.2
Process Ranges (Mean ± SD)
1)
Town Branch WWTP
Mixed Liquor Influent
Mixed Liquor Effluent
Digesters
218 ± 11.0
152 ± 16.8
3270.2 ± 245.8
4
SOP – Alkalinity
LFUCG Laboratory
Page 5 of 219
Revision Number 2
Last Revised 9/09
2)
11.
214 ± 27.1
177 ± 10.7
170 ± 15.8
284 ± 26.2
Data Analysis and Calculations
11.1
11.2
12.
West Hickman WWTP
Raw Effluent
Zone #1 Effluent
Zone #2 Effluent
Return Activated Sludge
Calculations determined by dilution.
See following pages for alkalinity concentration tables.
Bibliography
12.1
U.S. EPA. Method 310.1. Methods for Chemical Analysis of Water and Wastes.
EPA-600-4-79-020. U.S. Environmental Protection Agency; Office of Research
and Development, Washington, DC, 1982.
12.2
Standard Methods 2320-B. APHA-American Public Health Association Standard
Methods for the Examination of Water and Wastewater; 21th edition ed.;
American Water Works Association and Water Pollution Control Federation:
Washington, DC, 2005.
12.3
Total Alkalinity Measurement in Natural Waters. Application Information
Procedure 517. Thermo Fisher Scientific Inc.
http://www.thermo.com/com/cda/products/product_application_details/1,,11636,0
0.html
5
SOP – Alkalinity
LFUCG Laboratory
Page 6 of 219
Revision Number 2
Last Revised 9/09
ORION Total Alkalinity Test Kit
LOW RANGE 0 to 25 mg/L as CaCO3
Add 1 mL of ORION Total Alkalinity Reagent to 100 mL and mix well. Measure pH of mixture
and read total alkalinity from this table.
Observed
pH
Total
Alkalinity
Observed
pH
Total
Alkalinity
Observed
pH
Total
Alkalinity
Observed
pH
Total
Alkalinity
Observed
pH
Total
Alkalinity
4.00
0.0
4.35
5.4
4.70
10.9
5.05
16.4
5.40
21.9
4.01
0.1
4.36
5.6
4.71
11.1
5.06
16.6
5.41
22.1
4.02
0.3
4.37
5.8
4.72
11.3
5.07
16.7
5.42
22.2
4.03
0.4
4.38
5.9
4.73
11.4
5.08
16.9
5.43
22.4
4.04
0.6
4.39
6.1
4.74
11.6
5.09
17.1
5.44
22.6
4.05
0.7
4.40
6.2
4.75
11.7
5.10
17.2
5.45
22.7
4.06
0.9
4.41
6.4
4.76
11.9
5.11
17.4
5.46
22.9
4.07
1.1
4.42
6.5
4.77
12.0
5.12
17.5
5.47
23.0
4.08
1.2
4.43
6.7
4.78
12.2
5.13
17.7
5.48
23.2
4.09
1.4
4.44
6.9
4.79
12.4
5.14
17.8
5.49
23.3
4.10
1.5
4.45
7.0
4.80
12.5
5.15
18.0
5.50
23.5
4.11
1.7
4.46
7.2
4.81
12.7
5.16
18.2
5.51
23.7
4.12
1.8
4.47
7.3
4.82
12.8
5.17
18.3
5.52
23.8
4.13
2.0
4.48
7.5
4.83
13.0
5.18
18.5
5.53
24.0
4.14
2.1
4.49
7.6
4.84
13.1
5.19
18.6
5.54
24.1
4.15
2.3
4.50
7.8
4.85
13.3
5.20
18.8
5.55
24.3
4.16
2.5
4.51
8.0
4.86
13.4
5.21
18.9
5.56
24.4
4.17
2.6
4.52
8.1
4.87
13.6
5.22
19.1
5.57
24.6
4.18
2.8
4.53
8.3
4.88
13.8
5.23
19.3
5.58
24.7
4.19
2.9
4.54
8.4
4.89
13.9
5.24
19.4
5.59
24.9
4.20
3.1
4.55
8.6
4.90
14.1
5.25
19.6
5.60
25.1
4.21
3.2
4.56
8.7
4.91
14.2
5.26
19.7
4.22
3.4
4.57
8.9
4.92
14.4
5.27
19.9
4.23
3.6
4.58
9.1
4.93
14.5
5.28
20.0
4.24
3.7
4.59
9.2
4.94
14.7
5.29
20.2
4.25
3.9
4.60
9.4
4.95
14.9
5.30
20.4
4.26
4.0
4.61
9.5
4.96
15.0
5.31
20.5
4.27
4.2
4.62
9.7
4.97
15.2
5.32
20.7
4.28
4.3
4.63
9.8
4.98
15.3
5.33
20.8
4.29
4.5
4.64
10.0
4.99
15.5
5.34
21.0
4.30
4.31
4.7
4.8
4.65
4.66
10.2
10.3
5.00
5.01
15.6
15.8
5.35
5.36
21.1
21.3
4.32
4.33
5.0
5.1
4.67
4.68
10.5
10.6
5.02
5.03
16.0
16.1
5.37
5.38
21.5
21.6
4.34
5.3
4.69
10.8
5.04
16.3
5.39
21.8
6
SOP – Alkalinity
LFUCG Laboratory
Page 7 of 219
Revision Number 2
Last Revised 9/09
ORION Total Alkalinity Test Kit
HIGH RANGE 0 to 225 mg/L as CaCO3
Add 10 mL of ORION Total Alkalinity Reagent to 100 mL and mix well. Measure pH of
mixture and read total alkalinity from this table.
Observed
pH
Total
Alkalinity
Observed
pH
Total
Alkalinity
Observed
pH
Total
Alkalinity
Observed
pH
Total
Alkalinity
Observed
pH
Total
Alkalinity
3.66
0.0
4.01
47
4.36
93
4.71
140
5.06
187
3.67
1.2
4.02
48
4.37
95
4.72
141
5.07
188
3.68
2.5
4.03
49
4.38
96
4.73
143
5.08
190
3.69
3.8
4.04
51
4.39
97
4.74
144
5.09
191
3.70
5.2
4.05
52
4.40
99
4.75
145
5.10
192
3.71
6.5
4.06
53
4.41
100
4.76
147
5.11
194
3.72
7.8
4.07
55
4.42
101
4.77
148
5.12
195
3.73
9.2
4.08
56
4.43
103
4.78
149
5.13
196
3.74
11
4.09
57
4.44
104
4.79
151
5.14
198
3.75
12
4.10
59
4.45
105
4.80
152
5.15
199
3.76
13
4.11
60
4.46
107
4.81
153
5.16
200
3.77
15
4.12
61
4.47
108
4.82
155
5.17
202
3.78
16
4.13
63
4.48
109
4.83
156
5.18
203
3.79
17
4.14
64
4.49
111
4.84
157
5.19
204
3.80
19
4.15
65
4.50
112
4.85
159
5.20
206
3.81
20
4.16
67
4.51
113
4.86
160
5.21
207
3.82
21
4.17
68
4.52
115
4.87
161
5.22
208
3.83
23
4.18
69
4.53
116
4.88
163
5.23
210
3.84
24
4.19
71
4.54
117
4.89
164
5.24
211
3.85
25
4.20
72
4.55
119
4.90
165
5.25
212
3.86
27
4.21
73
4.56
120
4.91
167
5.26
214
3.87
28
4.22
75
4.57
121
4.92
168
5.27
215
3.88
29
4.23
76
4.58
123
4.93
169
5.28
216
3.89
31
4.24
77
4.59
124
4.94
171
5.29
218
3.90
32
4.25
79
4.60
125
4.95
172
5.30
219
3.91
33
4.26
80
4.61
127
4.96
173
5.31
220
3.92
35
4.27
81
4.62
128
4.97
175
5.32
222
3.93
36
4.28
83
4.63
129
4.98
176
5.33
223
3.94
37
4.29
84
4.64
131
4.99
177
5.34
224
3.95
3.96
3.97
3.98
39
40
41
43
4.30
4.31
4.32
4.33
85
87
88
89
4.65
4.66
4.67
4.68
132
133
135
136
5.00
5.01
5.02
5.03
179
180
181
183
5.35
226
3.99
44
4.34
91
4.69
137
5.04
184
4.00
45
4.35
92
4.70
139
5.05
186
7
SOP – NH3
LFUCG Laboratory
Page 8 of 219
Revision Number 1
Last Revised 09/09
Nitrogen, Ammonia
HACH Method Salicylate Method 10205
TNT+ 830, ULR (0.015 to 2.000 mg/L NH3–N)
TNT+ 831, LR (1 to 12 mg/L NH3–N)
TNT+ 832, HR (2 to 47 mg/L NH3–N)
EPA Method 350.1
1.
Scope, Significance to Process and Application
1.1
2.
3.
Ammonia concentrations in wastewater samples are an indication of nutrient
levels in the wastewater process stream. The reduction of ammonia levels
throughout the wastewater treatment process is highly important as plant effluent
nutrient concentrations must be low enough (See Sec. 10.1 Permit Requirements)
so as to avoid detrimental effect on the receiving environment. Low-level
ammonia nitrogen may be present in water naturally as a result of the biological
decay of plant and animal matter. Higher concentrations may be found in raw
sewage and industrial effluents. High concentrations in surface waters can
indicate contamination from waste treatment facilities, industrial effluents or
fertilizer runoff. Excessive ammonia concentrations are toxic to aquatic life, and
can exert an undesirable oxygen demand on the receiving stream.
Summary of Method
2.1
Executive Summary
Ammonium ions react at pH 12.6 with hypochlorite ions and salicylate ions in the
presence of sodium nitroprusside as a catalyst to form indophenol. The amount of
color formed is directly proportional to the ammonia nitrogen present in the
sample. Test results are measured at 690 nm. Nitrogen Ammonia analysis at
Town Branch Laboratory refers to the spectrophotometric analysis of nitrogen
ammonia compounds in a water/wastewater sample.
2.2
Discussion
This method covers the determination of ammonia-nitrogen exclusive of total
nitrogen, in drinking, surface and saline waters, domestic and industrial wastes.
ULR HACH method covers the range from 0.015 to 2.000 mg/L NH3-N. The
samples are analyzed colorimetrically with a HACH DR/5000.
Health & Safety Precautions
3.1
3.2
Watch out for broken glass from beakers and cylinders.
Wastewater samples have the potential to be hazardous, use appropriate caution.
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LFUCG Laboratory
Page 9 of 219
Revision Number 1
Last Revised 09/09
4.
Sample Handling and Preservation
4.1.
4.2.
4.3.
4.4.
4.5.
4.6.
5.
Reagents and Lab Equipment
5.1.
5.2.
5.3.
5.4.
5.5.
5.6.
5.7.
5.8.
5.9.
5.10.
5.11.
5.12.
6.
Collect samples in clean plastic or glass bottles. Best results are obtained with
immediate analysis.
Preserve the samples by reducing the pH to 2 or less with at least 2 mL of
Hydrochloric Acid.
Store at 4 °C (39 °F) or less.
Preserved samples may be stored up to 28 days.
Before analysis, warm stored samples to 20–23 °C (68–73.4 °F) and neutralize to
pH 7.0 with 5.0 N Sodium Hydroxide.
Correct the test result for volume additions.
Ammonia, TNTplus ULR Reagent Set (HACH TNT830)
Ammonia, TNTplus LR Reagent Set (HACH TNT831)
Ammonia, TNTplus HR Reagent Set (HACH TNT832)
Pipetors (100-1000 µL, 1–5 mL)
Pipet Tips
Nitrogen Ammonia Standard Solution, 1.0-mg/L NH3–N (HACH 189149)
Nanopure Water
Hydrochloric Acid Standard Solution, 1N
Hydrochloric Acid, concentrated ACS
Sodium Hydroxide Standard Solution, 1N (HACH 104532)
Test Tube Rack for 13-mm vial
DRB200 Reactor, 115 V, 9x13mm
Interferences
6.1.
6.2.
6.3.
6.4.
6.5.
The ions listed in the Interfering substances table have been individually tested up
to the given concentrations and do not cause interference. The cumulative effects
of these ions or the influence of other ions have not been determined.
Primary amines are determined and cause high-bias results. A 10,000-fold excess
of urea does not interfere. All reducing agents interfere and cause low-bias results.
Important Note: An analyte concentration greatly in excess of the stated range
will adversely affect color formation, resulting in a false reading within the
method range.
Measurement results can be verified using sample dilutions or standard additions.
Samples with severe interferences require distillation. Perform the distillation
procedure using the HACH General Purpose Distillation Set.
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LFUCG Laboratory
Page 10 of 219
Revision Number 1
Last Revised 09/09
7.
Procedures
7.1
TNT 830 ULR (0.015 to 2.000 mg/L NH3–N)
1) Carefully remove the protective foil lid from the DosiCap™ Zip. Unscrew the
cap from the vial.
2) Carefully pipet 5.0 mL of sample into the vial. Immediately proceed to step 3.
3) Flip the DosiCap Zip over so that the reagent side faces the vial. Screw the
cap tightly onto the vial.
4) Shake the capped vial 2–3 times to dissolve the reagent in the cap. Verify that
the reagent has dissolved by looking down through the open end of the
DosiCap Zip.
5) Wait 15 minutes.
6) After 15 minutes, invert the sample an additional 2–3 times to mix. The color
remains constant for an additional 15 minutes after the timer expires.
7) Thoroughly clean the outside of the vial with a Kim-wipe.
8) Insert the prepared vial into the DR5000 cell holder. Slide the lid closed. The
instrument reads the barcode, then selects and performs the correct test.
Results are in mg/L NH3–N.
7.2
TNT 831 LR (1 to 12 mg/L NH3–N)
1) Carefully remove the protective foil lid from the DosiCap™ Zip. Unscrew the
cap from the vial.
2) Carefully pipet 0.5 mL (500 μL) of sample into the vial. Immediately proceed
to step 3.
3) Flip the DosiCap Zip over so that the reagent side faces the vial. Screw the
cap tightly onto the vial.
4) Shake the capped vial 2–3 times to dissolve the reagent in the cap. Verify that
the reagent has dissolved by looking down through the open end of the
DosiCap Zip.
5) Wait 15 minutes.
6) After 15 minutes, invert the sample an additional 2–3 times to mix.
7) The color remains constant for an additional 15 minutes after the timer
expires.
8) Thoroughly clean the outside of the vial with a Kim-wipe .
9) Insert the prepared vial into the DR5000 cell holder. Slide the lid closed. The
instrument reads the barcode, then selects and performs the correct test.
Results are in mg/L NH3–N.
10
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LFUCG Laboratory
Page 11 of 219
Revision Number 1
Last Revised 09/09
7.3
TNT 832 HR (2 to 47 mg/L NH3–N)
1) Carefully remove the protective foil lid from the DosiCap™ Zip. Unscrew the
cap from the vial.
2) Carefully pipet 0.2 mL (200 μL) of sample into the vial. Immediately proceed
to step 3.
3) Flip the DosiCap Zip over so that the reagent side faces the vial. Screw the
cap tightly onto the vial.
4) Shake the capped vial 2–3 times to dissolve the reagent in the cap. Verify that
the reagent has dissolved by looking down through the open end of the
DosiCap Zip.
5) Wait 15 minutes.
6) After 15 minutes, invert the sample an additional 2–3 times to mix.
7) The color remains constant for an additional 15 minutes after the timer
expires.
8) Thoroughly clean the outside of the vial with a Kim-wipe.
9) Insert the prepared vial into the DR5000 cell holder. Slide the lid closed. The
instrument reads the barcode, then selects and performs the correct test.
Results are in mg/L NH3–N.
7.4
Reagent blanks
A reagent blank can be measured and the value subtracted from the results of each
test performed in same reagent lot. Use deionized water in place of sample in the
Salicylate method, TNTplus 830, 831, or 832 test.
To subtract the value of the blank from a series of measurements:
1. Measure the blank per step 3.
2. Turn on the reagent blank option.
3. The measured value of the blank should be displayed in the
highlighted box. Accept this value.
The reagent blank value will now be subtracted from all results until the function
is turned off or a different method is selected. Alternately, the blank can be
recorded and entered at any later time by pressing the highlighted box and using
the keypad to enter the value.
7.5
Sample blanks
Colored or turbid samples can cause high results. To compensate for color or
turbidity the procedure is repeated without the addition of the color forming
reagent that is present in the DosiCap Zip.
11
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LFUCG Laboratory
Page 12 of 219
Revision Number 1
Last Revised 09/09
To determine the sample blank:
1. Run the Salicylate method, TNTplus 830, 831, or 832 test, but do not
remove the foil from the DosiCap Zip in step 1.
2. Replace the cap in its original position in step 3.
3. Subtract the value obtained in step 8 from the value obtained on the
original sample to give the corrected sample concentration.
Samples without color or turbidity do not require sample blanks.
7.6
Helpful Hints
1) After washing used glassware, all glassware must be thoroughly rinsed with
lab grade water.
2) Analysis results are directly proportional to sample volumes therefore it is
very important that sample volume measurements are accurate.
3) Ammonia TNTplus tubes are temperature sensitive and must be stored in the
refrigerator (4o C) when not in use.
8.
9.
Standard Preparation
8.1.
ULR Low Standard (1.01 mg/L NH3–N)
Combine 1.00 mL of 10.1 mg/L standard + 9.0 mL DI water
8.2.
HR High Standard (10.1 mg/L NH3–N)
Add 20.0 mL of stock ammonia standard solution (100 ppm NH3–N) to 200 mL
volumetric flask. Dilute to 200 mL with Nanopure water.
QA/QC Requirements
9.1
9.2
9.3
A low standard (1.01 mg/L) and a high standard (10.1 mg/L) must be analyzed
with every analytical run.
5% of all samples must be run in duplicate. Duplicate concentration values should
agree within 5%.
Data acceptance criteria: Analysis values for Standards must agree within 10% of
the standard’s known value and duplicate values must agree within 5%. If these
criteria are not met, corrective action is indicated. See Quality Assurance Program
(QAP) Sec. 15 “Corrective Action Policies and Procedures”.
12
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LFUCG Laboratory
Page 13 of 219
Revision Number 1
Last Revised 09/09
10.
Expected Results
10.1
KPDES Permit Requirements
1) Town Branch Wastewater Treatment Plant effluent limitations:
Weekly average = 3 mg/L (May 1 - October 31)
Monthly average = 2 mg/L (May 1 - October 31)
Weekly average = 10.5 mg/L (November 1 - April 30)
Monthly average = 7 mg/L (November 1 - April 30)
2) West Hickman Wastewater Treatment Plant effluent limitations:
Weekly average = 6 mg/L (May 1 - October 31)
Monthly average = 4 mg/L (May 1 - October 31)
Weekly average = 15 mg/L (November 1 - April 30)
Monthly average = 10 mg/L (November 1 - April 30)
In the event that analysis results indicate values greater then KPDES permit
requirements, retest. If the value indicated by the retest is greater than KPDES
permit requirements, Immediately notify the Plant Superintendent and the
Laboratory Supervisor.
10.2
Process Ranges
Typical ammonia values vary a great deal throughout the wastewater treatment
process, depending on sample location and environmental conditions (i.e. high
rainfall events).
The following are typical Ammonia values found in process waters:
Raw Influent
0 mg/L to ≈ 25 mg/L
Plant Effluent 0 mg/L to 2.0 mg/L
11.
Data Analysis and Calculations
11.1
Concentrations are read directly from the DR5000 spectrophotometer. Ensure that
test results are corrected for volume dilutions.
13
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LFUCG Laboratory
Page 14 of 219
Revision Number 1
Last Revised 09/09
12.
Bibliography
12.1
U.S. EPA Method 350.1. Nitrogen, Ammonia, Colorimetric, Automated Phenate.
Methods for Chemical Analysis of Water and Wastes. EPA-600-4-79-020. U.S.
Environmental Protection Agency; Office of Research and Development,
Washington, DC, 1982.
12.2
HACH. DOC316.53.01081, Method 10205 Nitrogen-Ammonia Salicylate
method, TNTplus 830. HACH Company, Loveland, CO, 2008.
12.3
HACH. DOC316.53.01082, Method 10205 Nitrogen-Ammonia Salicylate
method, TNTplus 831. HACH Company, Loveland, CO, 2008.
12.4
HACH. DOC316.53.01083, Method 10205 Nitrogen-Ammonia Salicylate
method, TNTplus 832. HACH Company, Loveland, CO, 2008.
14
SOP – CBOD5
LFUCG Laboratory
Page 15 of 219
Revision Number 5
Last Revised 09/09
Biochemical Oxygen Demand (BOD5) and
Carbonaceous Biochemical Oxygen Demand (CBOD5) Analysis
EPA Method 405.1 (Editorial Revision 1974)
Standard Methods 5210-A and 5210-B
1.
Scope, Significance to Process and Application
The Biochemical Oxygen Demand (BOD5) and Carbonaceous Biochemical
Oxygen Demand (CBOD5) analysis assesses the concentration and general
composition of organic matter in raw water supplies, wastewaters, treated
effluents, and receiving waters. This test is used to determine the efficiency of the
treatment process at Town Branch, West Hickman, and Blue Sky Wastewater
Treatment Plants.
2.
3.
Summary of Method
2.1
Executive Summary
An initial dissolved oxygen (DO) reading is taken of the sample (or dilution). The
sample is then incubated in the dark at 20ºC for 5 days. A final DO reading is then
taken. Initial and final dissolved oxygen values are entered into a spreadsheet that
calculates the BOD and CBOD values which are expressed in milligrams per liter.
2.2
Discussion
The BOD5 test measures the amount of oxygen uptake caused by both the
biodegradation by micro-organisms of organic materials in wastewaters
(carbonaceous demand) and the oxidation of nitrogen forms in wastewaters
(nitrogenous demand) over a 5 day incubation period, coupled with calculations to
derive the Biochemical Oxygen Demand (BOD) of the sample tested. The CBOD5
test involves the addition of a nitrification inhibitor to exclude nitrogenous
demand allowing the measurement of just the carbonaceous demand (CBOD).
Health & Safety Precautions
3.1
4.
All municipal and industrial wastewaters are potentially hazardous.
Gloves and goggles should be worn when dispensing these samples.
Sample Handling and Preservation
4.1
4.2
4.3
Collect sample in plastic or glass and store at 4ºC.
Run analysis within 48 hours.
Grab samples:
If tested within 2 hours of collection, no cooling is needed.
If testing cannot start within 2 hours, cool to 4ºC and test within 6 hours.
15
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LFUCG Laboratory
Page 16 of 219
Revision Number 5
Last Revised 09/09
4.4
5.
Reagents
5.1
5.2
5.3
5.4
5.5
5.6
6.
Sodium Sulfite solution, 0.025 N
Instill 0.1575 g ± 0.0001 g Sodium sulfite into 100 mL volumetric flask and bring
to mark with Nanopure lab water (Note: This solution is not stable - prepare daily)
HACH BOD Nutrient Buffer Pillows, 3 L and 6 L
BOD Seed Inoculum (Polyseed®)
NCL Glucose-Glutamic Acid BOD Standard (198 ± 30.5 mg/L)
HACH Nitrification Inhibitor with dispenser cap
Nanopure laboratory water
Equipment & Lab Ware
6.1.
6.2.
6.3.
6.4.
6.5.
6.6.
6.7.
6.8.
6.9.
6.10.
6.11.
6.12.
6.13.
6.14.
7.
Composite samples:
Use the same criteria as for grab samples starting at the end of compositing.
Samples should be at 4ºC. At no time should an analyst test a 24 hour composite
sample after the 48 hour holding period.
300 mL disposable BOD bottles (Environmental Express)
BOD bottle stoppers and plastic caps
HACH HQ40d Portable Meter with LBOD101 probe
Control Co® Digital Barometer
Fisher Scientific® Isotemp Incubator Model 304 at 20ºC ± 1ºC
9 L glass dilution water bottle
Siphon hose (silver treated blue hose) and valve
Graduated cylinders to measure samples (250 and 500 mL)
Squeeze bottle for dispensing dilution water
Squeeze bottle with lab water to rinse probe between initial DO readings
500 mL and 1 L Erlenmeyer flasks
10 mL and 500 mL beakers
Barant Co® vacuum/pressure aeration system with hose and diffuser
Labsystems® adjustable 1-5 mL finnpipette
Interferences
7.1
7.2
7.3
7.4
Greases and oils in the sample.
Non representative particles such as leaves, sticks and debris.
Temperature differentials (i.e. ambient temperature DO probe used to analyze a
colder sample).
Air entrapped in the BOD bottle (bubbles).
16
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LFUCG Laboratory
Page 17 of 219
Revision Number 5
Last Revised 09/09
8.
Procedures
8.1
Steps
8.1.1
Preparation of BOD stoppers
1) BOD stoppers are washed with detergent and tap water.
2) Stoppers are then soaked in HCl (1:1) for at least 1 h.
3) Stoppers are then soaked in Nanopure water.
4) Allow to air-dry.
5) Wrap stoppers in blue sterile paper and indicator tape.
6) Autoclave wrapped stoppers.
7) Stoppers are maintained in sterile wrap until ready to use.
8.1.2
Preparation of BOD bottles
1) Obtain correct number of Env. Express disposable 300 mL BOD
bottles (Use only once).
2) Using a permanent marker, label bottles in accordance with bench
sheet.
3) To each sample BOD bottle add 2 shots (cycle the dispenser twice) of
HACH® Nitrification Inhibitor. Do not add Nitrification Inhibitor to
the unseeded blank, the blanks, or to industrial samples.
8.1.3
Preparation of dilution water
1) Place 9 L of Nanopure water into a 10 L dilution bottle.
2) Place filled dilution bottle on stir plate inside incubator (20 ± 1º C).
3) Add a 3 L pillow and a 6 L pillow of BOD nutrient buffer into the
dilution water bottle (1 mL nutrient to 1 L of lab water)
4) Mix thoroughly using a magnetic stirrer.
8.1.4
Preparation of BOD seed
1) Siphon 500 mL dilution water into 500 mL graduated cylinder.
2) Pour the 500 mL of dilution water into 1 L Erlenmeyer flask.
3) Pull apart Polyseed® capsule and pour contents into flask.
4) Submerge the aerator discharge diffuser in the flask using a support
stand to hold the diffuser centered at the bottom of the flask, turn the
aerator on.
5) Aerate contents of flask for at least one hour.
6) Decant the supernatant carefully into a clean 500 mL beaker so as not
to allow any bran in the remaining seed solution.
7) Stir using a spin bar and magnetic stir table.
8) The Polyseed® solution should be used within 6 hours of rehydration
of the capsule’s contents.
17
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LFUCG Laboratory
Page 18 of 219
Revision Number 5
Last Revised 09/09
8.1.5
Siphon dilution water into a labeled squirt bottle for dispensing.
8.1.6
Preparation of HACH HQ40d Meter with LBOD101 probe
1) Calibrate DO meter at the beginning of the analysis day (see SOP-DO,
Section 8).
2) Meter must be calibrated a minimum of once per analysis day.
Recalibrate meter if reading are taken in the afternoon.
3) Rinse LBOD probe between samples with Nanopure water.
8.1.7
Blank Control Samples
1) Prepare four BOD bottles as follows:
label one bottle “Unseeded Blank” - NO seed will be added
label one bottle “Blank 1” and add 2 mL of seed solution
label one bottle “Blank 2“ and add 3 mL of seed solution
label one bottle “Blank 3” and add 4 mL of seed solution
2) Siphon dilution water into BOD bottles until full.
3) Measure initial DO for each and record in bench sheet.
4) Replace any displaced dilution water, stopper, cap, and incubate at 20
± 1o C for 5 days.
5) The 5-day BOD at 20ºC should have a depletion of 0.6 to 1.2 mg/L.
8.1.8
Dilution water Quality Control (QC) – Unseeded Blank
1) Siphon 300 mL of dilution water into BOD bottle.
2) Take initial DO and record in bench sheet.
3) Replace any displaced dilution water, stopper, cap, and incubate at 20
± 1o C for 5 days.
4) The 5-day DO should be ± 0.2 mg/L of initial DO.
8.1.9
BOD standard preparation - Quality Control (QC) check
1) Obtain Glucose-Glutamic Acid BOD standard from the “Micro
Refrigerator” located under the fecal incubators.
2) Allow the standard to warm up to room temperature before using.
3) Using a sterile 10 mL pipette, add 6 mL BOD standard into BOD
bottle labeled “Standard”.
4) Siphon dilution water into BOD bottle until full.
5) Take initial DO and record in bench sheet.
6) Replace any displaced dilution water, stopper, cap, and incubate at 20
± 1o C for 5 days.
7) The 5-day BOD of the standard should be 198 mg/L ± 30.5 mg/L.
18
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LFUCG Laboratory
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Revision Number 5
Last Revised 09/09
8.1.10 Sample preparation
1) Place the sample volume indicated on the bench sheet into each
labeled sample BOD bottle.
2) Add 4 mL of BOD seed into each bottle.
3) Siphon dilution water into BOD bottle until full.
4) Read initial DO and record each sample on bench sheet.
5) Replace lost volume of BOD bottle with dilution water until full and
cap with stopper.
6) Place plastic caps over BOD bottles to protect water seal.
7) Rinse probe with lab water between samples.
8) Incubate BOD bottles at 20 ± 1º C for 5 days.
Check incubator temperature and adjust if needed. For each
incubator, record in the daily logs both the date and daily
temperatures, and initial. The daily logs are located next to BOD
station.
8.1.11 After 5-day incubation
1) Calibrate DO meter (see SOP-DO, Section 8).
2) Measure and record the 5-day DO values for each sample on the bench
sheet and turn in for data entry (see Section 11. Data Analysis and
Calculations).
8.2
9.
Helpful Hints
1) Samples that are caustic or acidic should be neutralized to pH 6.5 to 7.5.
2) Samples containing residual chlorine should have chlorine removed by adding
10 drops of 10% Sodium sulfite (Na2SO3) solution.
3) For samples that are known to have high concentrations of solids, make serial
dilutions in 100 mL volumetric flasks using Nanopure laboratory water so as
to yield a 40% to 60% DO uptake after 5 days.
QA/QC Requirements
9.1
9.2
9.3
9.4
9.5
Calibrate DO meter at the beginning of each analysis day. Recalibrate in the
afternoon.
Run BOD Standard each batch: 6 mL of 198 mg/L GGA standard.
Run 4 blanks with each batch (3 seeded, one unseeded).
Run 1 random duplicate per 20 samples.
Data acceptance criteria:
Analysis values for GGA Standards should be 198 mg/L ± 30.5 mg/L.
Duplicate values must agree within 5%.
Blank depletion values must be no greater than 0.2 mg/L.
If these criteria are not met, corrective action is indicated (See Quality Assurance
Program (QAP) Sec. 15 “Corrective Action Policies and Procedures”).
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10.
Expected Results
10.1
KPDES Permit Requirements
PTE: 10 mg/L maximum for monthly average and
15 mg/L maximum for weekly average.
In the event that analysis results indicate values greater then KPDES permit
requirements, notify the Plant Superintendent and Laboratory Supervisor.
10.2
11.
Process Ranges
Influent BOD values vary with season, rainfall amounts and influent flow rate.
Typical influent values for Town Branch Plant range 200 mg/L to 50 mg/L with
an average of approximately 130 mg/L.
West Hickman Plant influent values range from 250 mg/L to 50 mg/L with an
average of approximately 180 mg/L.
Typical effluent BOD values at both Town Branch and West Hickman
are <10 mg/L.
Data Analysis and Calculations
Initial and final dissolved oxygen values are entered into the BOD/CBOD worksheet
which calculates the BOD or CBOD values in accordance with the following
Given:
B1 = initial DO reading of blank
B2 = initial DO reading of blank
D1 = initial DO reading of sample
D2 = 5-day DO reading of sample
S = volume of seed used in blank
P = volume of seed per BOD sample bottle
V = volume of sample in BOD bottle
Then by Calculation:
(B1 - B2 ) = depletion of blank
(D1 - D2 ) = depletion of sample
(B1 - B2 /S) = DO used/ml of seed
F = seed correction = (B1 - B2 )/S*P)
C = (D1 - D2)-F = depletion corrected for seed
BOD or CBOD = C*(300/V)
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12.
Bibliography
12.1
U.S. EPA Method 405.1, Biochemical Oxygen Demand, 5 Days @ 20ºC, Issued
1971, Editorial revision 1974. Methods for Chemical Analysis of Water and
Wastes. EPA-600-4-79-020. U.S. Environmental Protection Agency; Office of
Research and Development, Washington, DC, 1982.
12.2
Standard Methods 5210-A. and 5210-B. Biochemical Oxygen Demand (BOD).
APHA-American Public Health Association Standard Methods for the
Examination of Water and Wastewater; 21th edition ed.; American Water Works
Association and Water Pollution Control Federation: Washington, DC, 2005.
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SOP – Residual Chlorine
LFUCG Laboratory
Page 22 of 219
Revision Number 4
Last Revised 09/09
Total Residual Chlorine Analysis
HACH AutoCAT 9000 Total Chlorine Amperometric Forward Titration
procedure equivalent to
EPA Method 330.3 Chlorine, Total Residual (Titrimetric, Amperometric), Issued 1978
Standard Methods Part 4500-Cl D. (Chlorine Residual Amperometric Titration Method)
1.
Scope, Significance to Process and Application
1.1
2.
3.
Disinfection by chlorination is considered to be the primary mechanism for the
inactivation/destruction of pathogenic organisms in wastewater treatment plant
effluents and to prevent the spread of waterborne diseases to downstream users
and the environment. Final clarifier effluent is treated with chlorine as enters the
chlorine contact tanks allowing contact time for disinfection to transpire. Final
effluent is then treated with a dechlorinating agent reducing chlorine residual
concentration to within acceptable limits (see Section 10.1 Permit Limits).
Complete dechlorination is necessary to prevent chlorine related adverse effects
on the receiving environment. Town Branch Waste Water Treatment Plant uses
Chlorine Dioxide (ClO2) for chlorination and Sulfur Dioxide (SO2) as the
dechlorinating agent. Residual Chlorine analysis of treated plant effluent
validates efficacy of dechlorinating agent dosing and permit compliance.
Summary of Method
2.1
Executive Summary
Town Branch Laboratory uses a HACH AutoCAT 9000 autotitrator to perform
Residual Chlorine determinations. The AutoCAT 9000 bench top system
automatically completes all USEPA- approved amperometric titration methods for
chlorine, calculates analyte concentration, and provides real-time graphics
display. The AutoCAT’s forward amperometric titration procedure has a range of
0.0012 mg/L to 5.0 mg/L with an estimated detection limit of 0.0012 mg/L
2.2
Discussion
Chlorine (hypochlorite ion, hypochlorous acid) and chloramines liberate iodine
from potassium iodide at pH 4 or less in stoichiometeric proportions. The iodine
is titrated with a reducing agent phenylarsine and an amperometer detects the
endpoint. Although the actual measurement is that of the samples oxidation
potential, it is calculated and expressed as mg/L Cl because chlorine is the
dominating oxidizing agent present.
Health & Safety Precautions
3.1
3.2
Glassware involved, possible cut hazard.
Wastewater samples have the potential to be hazardous, use appropriate caution.
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4.
Sample Handling and Preservation
4.1
4.2
4.3
5.
Reagents
5.1
5.2
5.4
5.5
6.
Reagent 1 - Potassium Iodide 5%
Reagent 2 - pH 4.00 Buffer (Certified Acetic Acid). Both reagents are located
adjacent to the AutoCAT unit
Phenylarsine Oxide Standard Solution 0.00564 N
Chlorine Standard Solution, 25-30 mg/L as Cl2
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
7.
Residual chlorine is subject to dissipation by exposure to sunlight, mechanical
agitation, exchange of gases with the atmosphere and reaction with compounds in
the wastewater over time. For these reasons chlorine residuals should be analyzed
immediately (within 15 minutes of sampling time).
The sample should be taken gently into a glass 300 mL BOD bottle, completely
filling to above the base of the neck and installing the tapered glass stopper in a
manner that precludes air bubbles in the sample.
All glassware used in this method must have no chlorine demand, therefore do not
use plastic containers and pre-treat glassware accordingly. To remove chlorine
demand from clean glassware, soak in a dilute bleach solution (1 mL commercial
bleach to 1 liter of Nanopure water) for at least one hour. After soaking, rinse
thoroughly with Nanopure water. After analysis, thoroughly rinse all glassware
with Nanopure water to reduce the need for pretreatment.
HACH AutoCAT 9000 - Chlorine Amperometric Titrator
Beakers 250 mL
Graduated Cylinders 250 mL
1 mL fixed volume Finnpipette and 1 mL tips
Stirring bars.
Interferences
7.1
7.2
7.3
7.4
7.5
Accurate determinations of free chlorine cannot be made in the presence of
Nitrogen trichloride or Chlorine dioxide.
Some organic chloramines can also interfere.
Free halogens other than chlorine also will titrate as free chlorine.
Interference from copper has been noted in samples taken from copper pipe or
after heavy copper sulfate treatment of reservoirs.
Contamination of probe by metal ions such as copper, silver, iron interfere with
amperometric titrations. Fouled electrodes will not produce sharp endpoints.
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7.6
7.7
8.
Extended sample hold times, volatilization from mechanical agitation, and
exposure to various light sources can affect results.
At very low temperatures, there is slow response of cell and longer time is
required, but precision is not compromised.
Procedures
8.1
Steps
1) Prior to testing, pre-rinse all glassware and stir bars with sample (Do not rinse
with Nanopure once pre-rinsed).
2) Using a 250 mL graduated cylinder measure 200 mL of sample.
3) Pour sample into a 250 mL beaker with stirring bar, raise the electrode
assembly and place the beaker on unit.
4) Turn on instrument. The display will request user to press “1” for “Total Cl2
Fwd”, press “1”, display will then request user confirmation, press “1” to
confirm.
5) The display will request confirmation of sample volume (200 mL) press “1”
6) The sample will begin to stir.
7) Display will request the addition of 1 mL of Potassium Iodide 5% (Reagent 1),
pipette reagent into sample, then press “OK”
8) Display will request the addition of 1 mL of Acetate buffer pH 4 (Reagent 2),
pipette reagent into sample, then press “OK”
9) A mixing timer will countdown for 5 sec. then the display will request that the
electrodes be dipped into the sample, lower electrode assembly into sample and
press “OK”.
10) The display will request user to confirm the “Increment Setting” (should be
0.0010), press “1” to confirm.
11) Unit will begin analysis; total time required for analysis will vary with sample
strength and chosen increment value. During analysis the display graphs the
progress of the titration. Upon completion the unit will display the resulting
concentration value and calculated confidence limits, press “OK” to accept
results.
12) Record the results on the Total Chlorine Residual bench sheet. In the case of
the PTE sample, also log (in the provided location) the time sampled, time
received, and time analysis began. Note: If sample hold time (time sampled to
time analysis begins) exceeds 15 minutes the analysis is void and must be
rerun, beginning with resampling.
13) Select “END” if done with analysis or “Continue” to proceed to the next
sample to be analyzed.
Note: More detailed general information on the AutoCAT unit can be found in the
operator’s manual with details on the Forward Amperometric procedure starting on
page 101. The manual is located on the shelf adjacent to the AutoCAT unit.
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9.
8.2
Chlorine Standard Analysis
1) Obtain a Chlorine Standard Solution (25-30 mg/L) ampoule from “Chemical
Storage” fridge.
2) Carefully break top off ampoule.
3) Pipette 1.0 mL of standard into 1000 mL volumetric flask.
4) Bring to 1000 mL with Nanopure water.
5) Measure 200 mL of standard solution into beaker with stir bar.
6) Run titration as indicated in Section 8.1.
7) Record results in bench sheet.
8) The Laboratory Supervisor will determine if the standard is within the
expected range.
9) Measure 200 mL of Nanopure water into a clean 250 mL beaker and analyze
as indicated in Section 8.1 (This will be a Blank to confirm no chlorine carryover). Record results in bench sheet.
10) If chlorine is detected, re-run Blank until Below Detection Limit (BDL) is
obtained.
8.3
Helpful Hints
1) Analysis results are directly proportional to sample volumes therefore it is
very important that sample volume measurement is accurate.
2) Clean conditioned electrodes are required for the production of sharp, well
defined endpoints that are needed for precise analysis. Rinse electrodes
thoroughly before and after each use with Nanopure water, and store in
Nanopure water.
3) Routine use of the “Electrode Cleaning and Conditioning” procedure as
described in Section 9.1.4 of the Operator’s Manual will prevent problems.
4) Glassware must be clean and free of chlorine demand see section 4.3
QA/QC Requirements
9.1
A diluted standard (25-30 mg/L) and Blank(s) must be run once a week (See
Section 8.2).
9.2
5% of all samples must be run in duplicate.
9.3
Data acceptance criteria:
1) Results for the Standard must agree within 10% of the standard’s known
value.
2) Duplicate values must agree within 5%.
3) If these criteria are not met, corrective action is indicated. See Quality
Assurance Program (QAP) Sec. 15 “Corrective Action Policies and
Procedures”.
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10.
11.
Expected Results
10.1
KPDES Permit Requirements
KPDES Permit Limits on plant effluent residual chlorine at Town Branch WWTP
is a maximum monthly average of 0.010 mg/L, with a daily maximum limitation
of 0.019 mg/L. In the event that analysis results indicate values greater then
KPDES permit requirements, retest. If the value indicated by the retest is greater
than KPDES permit requirements, Immediately notify the Plant Superintendent
and Laboratory Supervisor.
10.2
Process Ranges
Expected residual chlorine results on plant effluent samples will be less than
0.010 mg/L, typically the results are BDL (below detection limit).
Data Analysis and Calculations
11.1
11.2
12.
Concentration values are read directly from the AutoCAT unit’s display, all
calculations are preformed internally.
The Laboratory Supervisor will determine if the results for the standard are within
the expected range.
Bibliography
12.1
U.S. EPA Method 330.3 Chlorine, Total Residual (Titrimetric, Amperometric)
Issued 1978. Methods for Chemical Analysis of Water and Wastes. EPA-600-479-020. U.S. Environmental Protection Agency; Office of Research and
Development, Washington, DC, 1982.
12.2
Standard Methods 4500-Cl D. Chlorine Residual Amperometric Titration Method.
APHA-American Public Health Association Standard Methods for the
Examination of Water and Wastewater; 21th edition ed.; American Water Works
Association and Water Pollution Control Federation: Washington, DC, 2005.
12.3
HACH AutoCat 9000 Chlorine Amperometric Titrator Instruction Manual.
HACH Company, Loveland, CO.
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SOP – Free Cyanide
LFUCG Laboratory
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Revision Number 3
Last Revised 09/09
Free Cyanide CN (F)
HACH Method 8027 Pyridine-Pyrazalone Method
1.
Scope, Significance to Process and Application
1.1
2.
Measurement of the concentration of free cyanide in industrial wastewater
samples.
Summary of Method
2.1
Discussion
The Pyridine-Pyrazalone method used for measuring cyanide gives an intense blue color
with free cyanide.
3.
Health & Safety Precautions
3.1
3.2
3.3
4.
Sample Handling and Preservation
4.1
5.
Collect samples in glass or plastic bottles and analyze as quickly as possible. The
presence of oxidizing agents, sulfides and fatty acids can cause the loss of cyanide
during sample storage. Samples containing these substances must be pretreated as
described in the following procedures before preservation with sodium hydroxide.
If the sample contains sulfide and is not pretreated, it must be analyzed within 24
hours.
Reagents
5.1
6.
All municipal and industrial wastewaters are potentially hazardous. Gloves and
safety glasses should be worn when dispensing these samples.
Cyanides, their solutions, and Hydrogen cyanide liberated by acids are very
poisonous. Both gas and solutions can be absorbed through the skin. Latex gloves
and safety glasses should be used.
CyaniVer 3, CyaniVer 4, and CyaniVer 5 reagent powder pillows are used. May
be respiratory hazard.
CyaniVer 3, CyaniVer 4, and CyaniVer 5 reagent powder pillows.
Equipment & Lab Ware
6.1
6.2
6.3
Plastic beakers, at least 50 mL
Funnel, plastic or glass
Glass Microfibre Filter paper, 125mm diameter
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7.
6.4
6.5
Two 1-inch sample cells, 10-mL with cap (zeroing vial/sample vial)
Fixed 5 mL pipette with 5 mL pipette tip
6.6
6.7
Spec Color Standards kit
DR/4000 1-inch Cell Adapter
6.8
HACH DR/4000 Spectrophotometer
Interferences
7.1
7.2
7.3
7.4
7.5
8.
Chlorine.
Metals.
Oxidizing agents.
Reducing agents.
Turbidity
Procedures
8.1
Steps
8.1.1
Filtration
1) Pour approximately 40 mL from the Cyanide sample container to the
plastic beaker as soon as the sample arrives. Preserve the remaining
sample with 1.25 mL of 12.5 N NaOH for total cyanide analysis.
2) Fold the glass microfibre filter paper to fit in the funnel.
3) Slowly pour sample into filter so it does not overflow.
4) Collect filtered sample.
8.1.2
Reaction
1) Fill sample cell with 10 mL of filtered sample.
2) Add CyaniVer 3 powder pillow to the 10 mL in sample cell, cap and
shake 30 seconds. Wait an additional 30 seconds leaving the sample
undisturbed.
3) Add CyaniVer 4 powder pillow. Shake for 10 seconds and
immediately proceed to next step.
4) Immediately add CyaniVer 5 powder pillow. Shake the cell vigorously
for 15 seconds.
5) Set timer for 30 minutes.
6) If there is any cyanide present the sample will turn blue.
8.1.3
Using the DR4000 spectrophotometer on positive reaction
1) Fill a round sample cell with 10 mL of filtrated sample; this will be
used as the BLANK.
2) Touch HACH Programs on keypad and select program “1750
Cyanide”.
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Last Revised 09/09
3) Place BLANK sample into DR4000 cell holder. Close the light shield.
Touch “Zero” to zero instrument.
4) Place the prepared sample into the cell holder. Close the light shield.
Results will appear as mg/L cyanide.
8.2
9.
QA/QC Requirements
9.1
10.
Helpful Hints
1) Make sure that the CyaniVer 5 powder pillow is added immediately
after the 10 second shaking period from the previous reagent.
2) If interference is present, the sample will turn into a cloudy, murky
solution.
Use free cyanide standards.
Expected Results
Every now and then, a positive reaction occurs. If it is higher than the limit posted
on the industrial waste laboratory report (red number), inform the Laboratory
Supervisor immediately.
10.1
11.
Data Analysis and Calculations
11.1
12.
KPDES Permit Requirements
Depends on the particular industry being sampled.
None required.
Bibliography
12.1
HACH DR/4000 Method 8027. HACH Company, Loveland, CO.
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LFUCG Laboratory
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Revision Number 3
Last Revised 09/09
Total Cyanide CN (T)
HACH Method 8027 Pyridine-Pyrazalone Method with Distillation
Lachat Instruments Method 10-204-00-1-X (MICRO DIST Cyanide-1)
1.
Scope, Significance to Process and Application
1.1
2.
Measurement of the concentration of total cyanide in industrial waste water
samples.
Summary of Method
2.1
Discussion
The Pyridine-Pyrazalone method used for measuring cyanide gives an intense blue color
with free cyanide.
3.
Health & Safety Precautions
3.1
3.2
3.3
3.4
4.
Sample Handling and Preservation
4.1
5.
All municipal and industrial wastewaters are potentially hazardous. Gloves and
safety glasses should be worn when dispensing these samples.
Cyanides, their solutions, and Hydrogen cyanide liberated by acids are very
poisonous. Both gas and solutions can be absorbed through the skin. Latex gloves
and safety glasses should be used.
Industrial waste samples, CyaniVer 3, CyaniVer 4, and CyaniVer 5 reagent
powder pillows are used. Also used during preparation for the distillation process
is 0.75 mL of 7.11M sulfuric acid/0.79M magnesium chloride solution. Latex
gloves and safety glasses should be used.
During the distillation process the use of heat resistant gloves and safety glasses
are required.
Collect samples in glass or plastic bottles. Preserve the sample with 1.25 mL of
12.5N NaOH for analysis within 14 days of collection.
Reagents
5.1
5.2
5.3
5.4
CyaniVer 3, CyaniVer 4, and CyaniVer 5 reagent powder pillows
Releasing agent (7.11M sulfuric acid/0.79M magnesium chloride solution)
Trapping solution (0.950M standardized NaOH)
2.5N HCl
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6.
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9
6.10
6.11
7.
Lachat Micro Dist heating block
Lachat Micro Dist collector tubes, membranes and caps
Seal press
1-inch sample cells for as many sample as being analyzed
Fixed 0.580 mL pipette and tip
Adjustable 3.00 mL pipette and tip
Squirt bottle with Nanopure water
0.75 mL and 1.59 mL repipetor bottles
Spec Color Standards kit
HACH DR/4000 1-inch Cell Adapter
HACH DR/4000 Spectrophotometer
Interferences
7.1
7.2
7.3
7.4
7.5
8.
Chlorine.
Metals.
Oxidizing agents.
Reducing agents.
Turbidity
Procedures
8.1
Steps
8.1.1 Distillation
1) Allow Micro Dist heater block to warm up to 120o C.
2) Add 1.59 mL of trapping solution in each collector tube.
3) Cap tube with membrane.
4) Add 6 mL of sample in sample tube and add 0.75 mL releasing agent
(0.95M NaOH).
5) Immediately place the sample tube in the seal press. Seal the collector
tube to the sample tube using the pressing motion of the seal press.
6) Place tubes on the preheated block and set timer for 30 minutes.
7) When time is up remove the tube from the heating block and
immediately pull of the sample tube using a downward, twisting
motion. Allow 15 minutes for tubes to cool.
8) Rinse the walls of the collector tube. Slowly return the collector tube
to an upright position to gather all the droplets.
9) Break away the top half of the collector tube. Dilute to the 6 mL mark
with Nanopure water.
10) The distilled 6 mL sample in now ready to be analyzed.
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Last Revised 09/09
8.1.2 Reaction
1) Fill sample cell with the 6 mL distilled sample.
2) Add 0.580 mL of 2.5N HCl to sample cell, cap and shake for 10
seconds.
3) Add CyaniVer 3 powder pillow to the 10 mL in sample cell, cap and
shake for 30 seconds. Wait an additional 30 seconds leaving the
sample undisturbed.
4) Add CyaniVer 4 powder pillow. Shake for 10 seconds and
immediately proceed to next step.
5) Immediately add CyaniVer 5 powder pillow. Shake the cell vigorously
for 15 seconds.
6) Set timer for 30 minutes.
7) If there is any cyanide present the sample will turn blue.
8.1.3 Using DR4000 spectrophotometer on positive reaction
1) Use Spec Color Standards kit to test and zero the instrument.
2) Touch “HACH Programs” on DR4000 keypad and select program
“1750 Cyanide”.
3) Place BLANK sample into cell holder. Close the light shield. Touch
“Zero” to zero instrument.
4) Place the prepared sample into the cell holder. Close the light shield.
Results will appear as mg/L cyanide.
8.2
9.
QA/QC Requirements
9.1
10.
Helpful Hints
1) Make sure that the CyaniVer 5 powder pillow is added immediately
after the 10 second shaking period from the previous reagent.
2) If interference is present the sample will turn into a cloudy, murky
solution.
Use total cyanide standards.
Expected Results
Every now and then, a positive reaction occurs. If it is higher than the limit posted
on the industrial waste laboratory report (red number), inform the Laboratory
Supervisor immediately.
10.1
KPDES Permit Requirements
Depends on the particular industry being sampled.
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Revision Number 3
Last Revised 09/09
11.
Data Analysis and Calculations
11.1
12.
None required.
Bibliography
12.1
HACH DR/4000 Method 8027. HACH Company, Loveland, CO.
12.2
Lachat Micro Dist User Manual, Method Cyanide in Waters (MICRO DIST
Cyanide-1). Lachat Instruments, HACH Company, Loveland, CO.
12.3
U.S. EPA. Method 335.4. Revision 1.0, August 1993. Methods for Chemical
Analysis of Water and Wastes. EPA-600-4-79-020. U.S. Environmental
Protection Agency; Office of Research and Development, Washington, DC, 1982.
33
SOP – Cr 6+
LFUCG Laboratory
Page 34 of 219
Revision Number 3
Last Revised 09/09
Chromium, Total and Hexavalent (Cr 6+ )
TNT+ HACH Method 10219 (Chromium, Total)
TNT +HACH Method 10218 (Chromium, Hexavalent)
1, 5-Diphenylcarbohydrazide Method (0.03 to 1.00 mg/L Cr)
1.
Scope, Significance to Process and Application
1.1
2.
3.
Summary of Method
2.1
Executive Summary
Hexavalent chromium, if present in industrial wastewater samples, is measured by
HACH DR5000 spectrophotometer. Hexavalent chromium enters the water from
industrial wastes from metal plating facilities and from cooling towers where
chromate is used to inhibit corrosion.
2.2
Discussion
In the total chromium procedure, all chromium in the sample is oxidized to the
hexavalent chromium (Cr 6+). The hexavalent chromium then reacts with 1,5diphenylcarbazide to form 1,5-diphenylcarbazone. The amount of red color
formed with hexavalent chromium is directly proportional to the amount of
chromium present in the sample. Determine trivalent chromium by subtracting the
results of a separate hexavalent chromium test from the results of the total
chromium test. Test results are measured at 543 nm.
Health & Safety Precautions
3.1
3.2
4.
Measurement of the concentration of total and hexavalent (Cr 6+) in industrial
wastewater samples.
All municipal and industrial wastewaters are potentially hazardous. Gloves and
safety glasses should be worn when dispensing these samples.
Watch out for broken glass.
Sample Handling and Preservation
4.1
4.2
4.3
Collect samples in acid-washed glass or plastic containers.
To preserve samples for total chromium analysis, adjust the pH to 2 or less with
nitric acid (approximately 2 mL/L of the acid). Store preserved samples at 4 °C
for up to 6 months. Bring the sample temperature to 15–35 °C adjust the pH to
about 4 with 5.0 N NaOH before analysis.
To preserve samples for hexavalent chromium analysis, adjust the pH to 8 with
1N NaOH. Store at 4 °C for up to 24 hours. Bring sample to 15–35 °C. No pH
neutralization is required.
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Last Revised 09/09
5.
Reagents
5.1
5.2
5.3
5.4
6.
Chromium, Total and Hexavalent TNTplus Reagent Set (HACH TNT854)
Chromium, Trivalent, Standard Solution (50 mg/L Cr3+)
Chromium, Hexavalent Standard Solution (50 mg/L Cr6+)
Sodium Hydroxide, 1.0 N and 5.0 N
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
HACH DRB200 Reactor, 9x13 mm
Test Tube Rack
Variable volume pipette (1–5 mL) and tips
Plastic beakers or medicine cups
TNTplus Reactor Adapter Sleeves, 16-mm to 13-mm diameter
Funnel, plastic or glass
Glass Microfibre Filter paper, 125mm diameter
HACH DR5000 spectrophotometer
7.
Interferences
7.1
The ions listed in Interfering substances (See HACH method) have been
individually checked up to the given concentrations and do not cause interference.
Cumulative effects and the influence of other ions have not been determined.
7.2
Larger amounts of iron, copper and reducing and oxidizing agents give low-bias
results. Lead, mercury and tin give high-bias results.
7.3
Important Note: Undissolved chromium is not determined with the determination
of chromium(VI). An analyte concentration greatly (above 20 mg/L) in excess of
the stated range will adversely affect color formation, resulting in a false reading
within the method range.
7.4
Measurement results can be verified using sample dilutions or standard additions.
8.
Procedures
8.1
Steps
8.1.1 Filtration
1) Pour the sample from the sample container to the plastic beaker as
soon as the sample arrives.
2) Fold a glass microfibre paper to fit in the funnel.
3) Slowly pour sample into funnel so it does not overflow.
4) Collect filtrate.
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8.1.2
Method 10219 for Total Chromium
1) Turn on the DRB200 Reactor. Preheat to 100 °C. For DRB200
Reactors with 16-mm wells, make sure the 16-mm to 13-mm adapter
sleeve have been placed into each well before turning on the reactor.
2) Carefully remove the protective foil lid from the DosiCap™ Zip.
Unscrew the cap from the vial.
3) Pipet 2.0 mL of sample into the vial.
4) Turn the DosiCap Zip over so that the reagent side faces the vial.
Screw the cap tightly onto the vial.
5) Shake the capped vial 2–3 times to dissolve the reagent in the cap.
Make sure that the reagent has dissolved by looking down through the
open end of the DosiCap.
6) Heat the vial for one hour at 100 °C.
7) When the timer expires remove the hot vial from the reactor. Cool the
vials to 15–35 °C. Do not invert the vial after digestion.
8) Screw an orange DosiCap B onto the cooled vial.
9) Invert the vial 2-3 times to mix.
10) After inverting the tube, allow the vial to sit undisturbed for 2–3
minutes.
11) After the timer expires, invert the vial again 2–3 times.
12) Thoroughly clean the outside of the vial with a Kim-wipe. Insert the
prepared vial into the DR5000 cell holder. The instrument reads the
barcode, then selects and performs the correct test. Results are in mg/L
Cr.
13) No instrument Zero is required.
8.1.3
Method 10218 for Hexavalent Chromium
1) Remove cap and pipet 2.0 mL of sample into the vial.
2) Screw an orange DosiCap B on the vial.
3) Invert the vial 2-3 times to mix.
4) After inverting the tube, allow the vial to sit undisturbed for 2–3
minutes.
5) After the timer expires, invert the vial again 2–3 times.
6) Thoroughly clean the outside of the vial with a Kim-wipe.
7) Insert the prepared vial into the DR5000 cell holder. The instrument
reads the barcode, then selects and performs the correct test. Results
are in mg/L Cr6+.
8) No instrument Zero is required.
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8.2
9.
8.1.4
Reagent blanks
1) A reagent blank can be measured and the value subtracted from the
results of each test performed using the same reagent lot number. Use
deionized water in place of sample and run the procedure as described.
2) To subtract the value of the blank from a series of measurements,
measure the blank per step 12 of the total chromium procedure or step
7 of the hexavalent chromium procedure.
3) Press OPTIONS>MORE>REAGENT BLANK. Press ON. The
measured value of the blank should be displayed in the highlighted
box. Press OK to accept this value. The reagent blank value will now
be subtracted from all results until the function is turned off or a
different method is selected.
4) Alternately, the blank can be recorded and entered at any later time by
pressing the highlighted box and using the keypad to enter the value.
8.1.5
Sample blanks
1) Colored or turbid samples can cause high results. The digestion in the
total chromium procedure usually destroys all color and turbidity and a
sample blank is not required. To compensate for color or turbidity in
the determination of hexavalent chromium, the procedure is repeated
and the color forming reagent that is present in the DosiCap B is not
added.
2) To determine the sample blank for hexavalent chromium:
a) Run the procedure as written, but do not add the DosiCap B
Reagent in step 2.
b) Cap the vial with the original DosiCap Zip (do not remove the
foil).
c) The value obtained in step 7 is subtracted from the value obtained
on the original hexavalent chromium sample to give the corrected
sample concentration.
d) Alternatively, hexavalent chromium samples that contain turbidity
only may be filtered through a membrane filter and analyzed using
the hexavalent procedure. Results are reported as dissolved
hexavalent chromium.
3) Samples without color or turbidity do not require sample blanks.
Helpful Hints
1) Make sure to correct the test results for volume dilutions.
2) Wipe off any liquid or fingerprints from TNT+ tube.
QA/QC Requirements
9.1
None required.
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10.
11.
Expected Results
10.1
Report positive reactions to supervisor immediately.
10.2
KPDES Permit Requirements
Depends on the particular industry being sampled.
Data Analysis and Calculations
11.1
12.
None required.
Bibliography
12.1
12.2
12.3
HACH Water Analysis Handbook. Method 8023, Chromium Hexavalent (1,5Diphenylcarbohydrazide Method). 2nd Edition. 1992. HACH Company, Loveland,
CO.
HACH. DOC316.53.01035. Chromium, Total and Hexavalent. 1,5Diphenylcarbohydrazide, Method 10218 (Chromium, Hexavalent) and Method
10219 (Chromium, Total), TNTplus™ 854. 2008. HACH Company, Loveland,
CO.
Standard Methods Part 3500 B. Chromium, Colorimetric Method. APHAAmerican Public Health Association Standard Methods for the Examination of
Water and Wastewater; 21th edition ed.; American Water Works Association and
Water Pollution Control Federation: Washington, DC, 2005.
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Dissolved Oxygen Analysis (D.O.)
HACH Method 10360 Luminescent Dissolved Oxygen Probe Method
Proposed EPA Method 360.3 (Luminescence) for the Measurement of
Dissolved Oxygen in Water and Wastewater
1.
Scope, Significance to Process and Application
1.1.
1.2.
1.3.
1.4.
1.5.
2.
3.
The Dissolved Oxygen (D.O.) analysis is the measurement of the concentration of
oxygen dissolved in a water sample.
This method is recommended for samples containing intense color or turbidity.
This method is recommended for work in the field, as the equipment is portable,
allowing hold times to be minimized.
KPDES Permit Limits on Plant Effluent is a minimum of 7.0 mg/L.
Dissolved Oxygen concentration levels are very important in both process plant
effluent. In process, dissolved oxygen is required by various organisms and the
plant effluent dissolved oxygen levels must be conducive to the receiving
environment and within permit limits (See Section 10.1).
Summary of Method
2.1
Executive Summary
Dissolved Oxygen is measured directly by a HACH model HQ40d portable meter
and HACH model LBOD101 probe, located on the BOD bench. After the meter
indicates a stable reading the analyst records the value.
2.2
Discussion
The HACH LDO system uses a sensor coated with a luminescent material. Blue
light from an LED is transmitted onto the sensor surface, exciting the luminescent
material, which then emits red light as it relaxes. The presence of DO in the
process shortens the time it takes for the red light to be emitted. By measuring the
time lapse between when the blue light was transmitted and the red light is
emitted, a correlation is made to the concentration of DO in the effluent or other
solution. Between measurements, a red LED is used as an internal reference. The
measurement range for the method is 0.02 - 20.0 mg/L. The Method Detection
Limit (MDL; 40 CFR 136, Appendix B) has been determined as 0.05 mg/L and
the Minimum Level (ML; Reference 15.4) has been set at 0.20 mg/L.
Health & Safety Precautions
3.1
3.2
Glassware involved: possible cut hazard.
All municipal and industrial wastewaters are potentially hazardous.
Gloves and safety glasses should be worn when dispensing these samples.
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4.
Sample Handling and Preservation
4.1
4.2
4.3
5.
Reagents
5.1
6.
HACH HQ40d portable multi-meter
HACH Model LBOD101-01 probe with integrated stirrer
BOD bottles with 300 mL capacity and tapered ground glass stoppers
Sensor Cap replacements (HACH part # 5838000)
LDO Stirrer replacement kit (HACH part # 5850800)
Interferences
7.1
7.2
7.3
8.
Nanopure Lab Water
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
7.
Sample must be collected in a glass bottle (BOD bottle with stopper) filled to top,
with no airspace.
Sample must be analyzed immediately (15 minutes maximum on permit samples).
There is no applicable preservative with this method.
Salinity (salinity correction available, See Section 8.4.3 of the Users Manual).
Reactive gas: chlorine and hydrogen sulfide.
Air bubbles in sample or on surface of probe tip.
Procedures
8.1
Steps
8.1.1 Calibration and Start Up
It is recommended that the HQ40d Users Manual be initially consulted when
following these procedures. The manual is located in a yellow folder labeled “Lab
D.O. Meter”, in the yellow bin located adjacent to the meter.
1) Press the power button and allow the unit to perform its startup self check
routine.
2) Clean by rinsing with Nanopure lab water, then gently blot dry the probes tip
with a Kim-Wipe. Inspect the probe tip for indications contamination or
damage.
3) Take a 300 mL BOD bottle containing approximately one inch of lab water,
stopper and shake, remove stopper and replace it with the probe.
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4) Press Calibrate (blue button), the meter will prompt you to “Dry the probe and
place in water saturated air & press “Read”. Press the “Read” button, the
screen will scroll from 0 to 100%, then indicate “Calibration Complete”.
Record from display screen both the temperature and the dissolved oxygen
value (indicated under the temperature). Log the values on the dissolved
oxygen calibration section of the Dissolved Oxygen bench sheet under
Temperature and Dissolved Oxygen from HQ40d.
5) Note the barometric pressure value from the laboratory barometer (located
adjacent to the D.O. meter) and record on the dissolved oxygen calibration
section of the Dissolved Oxygen Bench sheet under “Barometer Reading”.
6) On the Lab computer, open the excel spreadsheet entitled “DO Meter
Calibration Sheet” and enter the barometric pressure, temperature and
dissolved oxygen values from the dissolved oxygen calibration section of the
Dissolved Oxygen Bench sheet. The spreadsheet will calculate the “Dissolved
Oxygen Calibration Point”, the “Dissolved Oxygen @ 1 ATM” and the Slope
%. Transfer the three values onto the dissolved oxygen calibration section of
the Dissolved Oxygen bench sheet, then print a copy of the spreadsheet and
file it in the yellow file (located adjacent to the D.O. meter).
7) Note the difference between the Dissolved Oxygen from HQ40d and the
Dissolved Oxygen Calibration Point - if it is greater than 0.2 mg/L, then the
calibration is not acceptable and must be repeated until criteria is met.
8) If sample measurements are made in the afternoon, the meter must be
calibrated again, due to changes in atmospheric pressure. Repeat steps in
Section 8.1.1.
8.1.2
Measurements
1) Make sure that the meter is properly calibrated.
2) Rinse the LBOD probe tip with Nanopure water.
3) Place the probe into BOD bottle, filled to the base of its neck with sample, and
turn on the self-contained stirring unit switch located on top of the probe.
Assure that there are no air bubbles on the surface of the probe tip.
4) Press “Read”, the screen will display “Stabilizing” and a progress bar will
scroll from 0 to 100%. Reading stability is indicated by the appearance of a
“Padlock” icon in the upper left corner of the display screen.
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5) Record the indicated value, turn the stirrer off, remove the probe and rinse tip
with Nanopure water and proceed to the next sample or store until needed.
6) DO NOT store probe in the BOD bottle containing water. Probe can be stored
dry on the bench top.
8.2
9.
QA/QC Requirements
9.1
9.2
9.3
10.
11.
Helpful Hints
1) The meter is designed to be maintenance free, when needed, clean the
exterior with a damp cloth.
2) The probe’s maintenance consist of maintaining the probe tip clean,
frequent rinsing with Nanopure water is sufficient.
3) DO NOT scrub the sensor cap or lens.
4) DO NOT use any organic solvents on the sensor cap or probe body.
Meter must be calibrated a minimum of once per analysis day.
Permit sample hold times must be 15 minutes or less.
Probe condition must be properly maintained through routine cleaning (See
Section 8.2, Helpful Hints).
Expected Results
10.1
KPDES Permit Requirements
7.0 mg/L is the lowest D.O. value allowable in a Plant Effluent sample at any
given time. In the event of a indicated value less than 7.0 mg/L, assure correct
calibration, resample, and retest. If the value indicated by retest is less than 7.0
mg/L. Immediately notify the Plant Superintendent and Laboratory Supervisor.
10.2
Process Ranges
Raw influent dissolved oxygen values are typically less than 1 mg/L.
Target values for mixed liquor dissolved oxygen concentration in the aeration
basins is 2.0 mg/L.
Data Analysis and Calculations
11.1
None required, values are taken directly when measurement stability is indicated.
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12.
Bibliography
12.1
Report on the Validation of Proposed EPA Method 360.3 (Luminescence) for the
Measurement of Dissolved Oxygen in Water and Wastewater. August 2004.
HACH Company, Loveland, CO.
12.2
Memorandum: EPA Recommendation for the use of HACH method 10360
[Revision 1.1, January 2006] (ATP Case # N04-0013).
12.3
HACH HQ Series Portable Meter Users Manual, September 2006, Edition 5.
HACH Company, Loveland, CO.
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Fecal Coliform - Membrane Filter Procedure
U.S. EPA 600/8-78-017 Microbiological Methods for Monitoring the Environment:
Water and Wastes
1.
2.
3.
Scope, Significance to Process and Application
1.1
The fecal coliform analysis is applicable to investigations of stream pollution, raw
water sources, and wastewater treatment systems.
1.2
The fecal coliform analysis differentiates between coliforms of fecal origin.
Summary of Method
2.1
Executive Summary
The sample is filtered through a Millipore® membrane filter. The filter is placed
on a filter pad containing media in a sterile Petri dish. The samples are then
incubated at 44.5°C ± 0.2°C for 24 hours ± 2 hours. Colonies are counted and
fecal coliform calculations are performed.
2.2
Discussion
Fecal coliforms are defined as gram-negative, non-spore forming rods. The major
species is Escherichia coli, which indicates fecal pollution and the presence of
enteric pathogens. Colonies produced by fecal coliform bacteria are various
shades of blue. Non-fecal coliform colonies are gray to cream colored.
Health & Safety Precautions
3.1
3.2
3.3
3.4
4.
All municipal and industrial wastewaters are potentially hazardous. Gloves and
safety glasses should be worn when dispensing these samples.
Possible exposure to enteric pathogens. Care must be taken to avoid undue
exposure.
A flame is used to sterilize forceps. Maintain the area around the flame clear.
Contaminated (used) Petri dishes and lab equipment must be placed in
Biohazardous waste container. This Biohazardous waste container is autoclaved
before disposal.
Sample Handling and Preservation
4.1
4.2
4.3
Samples should be collected in clean, sterile glass or plastic containers.
If chlorine is in the sample, containers should be treated with 4 drops of
10% Sodium thiosulfate before autoclaving.
Run test immediately after sampling, or preserve sample at 4°C for a maximum of
6 hours.
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5.
Reagents
5.1
5.2
5.3
5.4
5.5
6.
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9
6.10
6.11
6.12
6.13
6.14
6.15
6.16
6.17
6.18
7.
Peptone powder
Peptone buffer solution pH 7.00 ± 0.1 (stored at 4°C)
m-FC media with rosolic acid for fecal coliforms (stored at 4°C)
Sodium thiosulfate 10% solution
Lysol disinfectant, 20% solution
Vacuum flask
Millipore® single use 47 mm Petri dishes with pads
Millipore® sterilized 47 mm filter
Forceps
4.5 X 9 inch sterile sampling bags
Bunsen burner and striker
Pipettes and sterile tips
Sterilized filter holder (plastic or glass)
Gable topped water bath at 44.5°C ± 0.2°C
Thermometer
ASTM Thermometer
Tower Steam Indicator Strips
ODO-Clave® Heat Activated Deodorant Pads
Autoclavable Biohazard waste bags and deposit box
Autoclave
Sterile blue sheets
Indicator tape
Autoclavable Nalgene® squeeze bottles for peptone
Interferences
7.1
Bacteria from the surrounding environment.
7.2
Cross contamination from one sample to the next.
7.3
Lack of aseptic techniques.
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8.
Procedures
8.1
Steps
1) Clean work area with Lysol disinfectant, 20% solution.
2)
Light Bunsen burner with striker.
3)
Open sterile filter holder. Use sterile blue sheet as a sterile field. Indicator
trip and tape should indicate that the filter has been sterilized.
4)
Flame forceps and use it to remove the Indicator Strip without touching
anything else except the strip.
5)
Use Petri dishes with sterile pad already in dish.
6)
Break open ampule of media and pour onto media pad.
7)
Decant excess media and cover dish to protect sterile pad.
8)
Place the bottom of the sterile filter holder onto the vacuum flask.
9)
Flame forceps, remove sterilized filter from packaging and place onto
sterilized filter holder (grid side up). Do not touch the filter with anything
except the forceps.
10) Place or clamp the top unit onto filter holder.
11) Gently mix sample.
12) In advance, determine sample volume that will yield 20-60 fecal coliform
units (FCU).
13) If the volume of sample to be used is 0.1 to 5 mL, pour approximately 10
mL of peptone into filter unit before dispensing sample (Turn on vacuum
after the sample is introduced).
14) For sample volumes 5 to 50 mL, use sterile pipettes for dispensing into filter
unit.
15) Do not touch the inside of the filter holder unit. Do not allow the pipette tip
to touch the filter.
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16) For sample volumes 50 to 100 mL, pour sample into cylinder and use the
scale on the side of the cylinder for measurement.
17) Turn on vacuum.
18) Once sample has filtered through, turn off vacuum.
19) Rinse top of the filter unit with peptone two times using autoclaved peptone
in a Nalgene® squeeze bottle.
20) Turn on vacuum to drain peptone.
21) Turn off vacuum.
22) Flame forceps.
23) Remove top of the filter unit and place on the sterile blue field.
24) Open Petri dish.
25) Use sterile (flamed) forceps to grab the edge of the filter and remove it from
the filter holder unit.
26) Place filter, grid side up, onto edge of the Petri dish and gently slide it onto
the surface of the media saturated pad.
27) Replace Petri dish cover.
28) Place Petri dishes into a 4.5 X 9 inch sterile sampling bag. Make sure not
contaminate the inside of the bag.
29) Seal bag.
30) Place bag, with Petri dishes face down, into water bath at 44.5°C ± 0.2°C for
24 ± 2 hours.
31) Log initials, time, and date in the Microbiology bench sheet.
32) After 24 ± 2 hours, count blue colonies (See Section 10).
33) Log results, initials, time, and date in the Microbiology bench sheet.
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8.2
Before and After Blanks
1)
Blanks must be analyzed before and after filtration of a set of samples.
2)
Before any sample is filtered, place a sterile filter in the filter holder unit.
3)
Add 50 mL peptone buffer to filter unit.
4)
Turn on vacuum and filter the buffer, this will be the Before Blank.
5)
Remove and plate filter as indicated in Section 8.1.
6)
Proceed with sample filtration and plating.
7)
Run a Known Positive after all samples have been filtered (See Section 8.3).
8)
Place a sterile filter in the filter holder unit.
9)
Add 50 mL peptone buffer to filter unit.
10) Turn on vacuum and filter the buffer, this will be the After Blank.
11) Remove and plate filter as indicated in Section 8.1.
12) Log results, initials, time, and date in the Microbiology bench sheet.
8.3
Known Positive
1)
After all samples have been filtered, a Known Positive is filtered and plated
to ensure growth.
2)
Place a sterile filter in the filter holder unit.
3)
Add 10-20 mL peptone buffer to filter unit, then pipet 1.0 mL of mixed
liquor (or suitable sample with known fecal coliforms) into filter unit.
4)
Turn on vacuum and filter the sample, this will be the Known Positive.
5)
Remove and plate filter as indicated in Section 8.1.
6)
Log results, initials, time, and date in the Microbiology bench sheet.
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8.4
8.5
9.
Peptone Buffer Preparation
1)
Measure 1.0 g Peptone powder into 1L volumetric flask.
2)
Bring to volume with Nanopure water. Mix well.
3)
Pour Peptone buffer into Nalgene® autoclavable squeeze bottle.
4)
Take a sample of the buffer and measure pH, it should be 7.00 ± 0.1.
5)
Loosely screw caps/dispensers onto squeeze bottles.
6)
Autoclave Peptone buffer solutions.
7)
Allow to cool, then transfer squeeze bottles to Micro fridge.
Helpful Hints
1)
If the bacterial density of the sample is unknown, filter and plate out several
volumes or dilutions in order to achieve a countable density. The volumes
and/or dilutions should be expected to yield a countable membrane. In
addition, select two additional quantities representing one-tenth and ten
times this volume, respectively.
2)
Separate filter holder units may be required during a set of samples. These
will be indicated in the bench sheet.
3)
Do not use damaged or bent membrane filters.
4)
Rinse the filter unit thoroughly with Peptone buffer to avoid cross
contamination.
QA/QC Requirements
9.1
Before and After Blanks must be run with each set of samples tested.
9.2
One duplicate per test series must be run.
9.3
One “Known positive” must be run per test series.
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10.
Expected Results
10.1
KPDES Permit Requirements
200 CFU/100 mL for Monthly geometric mean (GED)
400 CFU/100 mL for Maximum Weekly GED
In the event that the GED is exceeds the KPDES permit requirements, notify the
Plant Superintendent and the Laboratory Supervisor.
10.2
11.
Data Analysis and Calculations
11.1
12.
Process Ranges
< 1 to >120,000 CFU/100 mL
See the following pages.
Bibliography
12.1
U.S. EPA. Microbiological Methods for Monitoring the Environment: Water and
Wastes. EPA 600/8-78-017. U.S. Environmental Protection Agency;
Environmental Monitoring and Support Laboratory, Office of Research and
Development, Washington, DC, 1978. Page 124.
12.2
Kentucky Department for Environmental Protection, Kentucky Division of Water
and the Kentucky Division of Compliance Assistance. Discharge Monitoring
Report Manual. 2009. August 10, 2009 revision. 28 pp.
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FECAL COLIFORM CALCULATIONS
The ways the answers are presented vary with the number of colonies on the plate.
A. Countable plates 1 with 20-60 Blue colonies.
Fecal Coliforms/100 mL =
Number of Blue colonies counted
Volume of sample filtered (mL)
x100
Example: 40 Blue colonies are counted and 50 mL of sample was used.
Fecal Coliforms/100 mL =
40
50
x100 = 80
B. Countable plates with less than 20 Blue colonies.
• If only one plate has been set-up, calculate as shown, but place an approximately equal to
(≈) sign 2 in front of the answer.
Fecal Coliforms/100 mL =
•
18
50
x100 = ≈ 36
If more than one plate has been set-up, calculate the total number of colonies from all the
plates and the total volume of sample used for all the plates. Use these totals to calculate
the number of coliforms per 100 mL.
Example:
10 Blue colonies
50 mL Sample
5 Blue colonies
40 mL Sample
3 Blue colonies
10 mL Sample
Fecal Coliforms/100 mL =
10 + 5 + 3
50 + 40 + 10
x100 = ≈ 18
C. Plates with no Blue colonies.
•
Do not use zero in the calculations. Place the number one in the equation and use the
largest volume of sample. Report as less than (<).
Example:
0 Blue colonies
0 Blue colonies
0 Blue colonies
50 mL Sample
40 mL Sample
10 mL Sample
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D. Countable plates with more than 60 Blue colonies.
•
If only one plate has been set-up, calculate as shown, but place a approximately equal to
(≈) sign in front of the answer.
Fecal Coliforms/100 mL =
•
95
50
x100 = ≈ 190
If more than one plate has been set-up, use the plate with the highest dilution (the lowest
sample volume). Place an approximately equal to (≈) sign in front of the answer.
Example:
150 Blue colonies
100 Blue colonies
80 Blue colonies
Fecal Coliforms/100 mL =
50 mL Sample
40 mL Sample
10 mL Sample
80
10
x100 = ≈ 800
E. Uncountable plates with more than 60 Blue colonies (TNTC).
•
•
If a sample contains colonies that are too numerous to count, conduct enough dilutions in
these tests to obtain discrete (“real number”) values. Reporting a non-numerical value,
such as TNTC, is unacceptable because you cannot average non-numerical values.
For samples in which subsequent dilutions do not produce a discrete value and are too
numerous to count, report results as 60,000. You would also use this value in the
geometric mean calculation. (KDEP DMR Manual, 2009).
F. To calculate the Geometric Mean (G.E.D.).
•
Zeros shall be recorded as < 1.
•
To calculate the logs of numbers with greater than, less than, and approximately equal to
sign, drop the sign. Do the math and then replace the sign as shown below.
•
If a column has only numbers without signs and numbers with less than signs, then the
total of this column will have a less than sign.
•
If a column has numbers without signs, numbers with less than signs, and numbers with
greater than sign, then the total of this column will have a less than sign.
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•
If a column has only numbers without signs and numbers with greater than signs, then the
total of this column will have a greater than sign.
•
If a column has any numbers with approximately equal to signs, then the total of this
column will have an approximately equal to sign.
Example:
Fecal Coliform/100 mL
Logarithm
100
50
<1
» 10
60
80
150
> 60000
2.00000
1.69897
0.00000
1.00000
1.77815
1.90309
2.17609
4.77815
< 83
15.33445
15.33445 / 8 = 1.91681
1.91681 = 83
1
Countable plate is a plate that has a number of well formed Blue colonies that can be easily
counted (0 to about 150). The EPA states, that a count of 20 to 60 colonies is the desired range
for Fecal Coliform plates.
2
Enter these signs as follows: The Font is Symbol. The size is 11.
Nomenclature
Approximately equal to
Greater Than
Less Than
Greater Than or equal to
Less Than or equal to
Symbols
»
>
<
≥
≤
Keystrokes
Hold down the Alt-Key and type 0187
Hold down the Shift-Key and type >
Hold down the Shift-Key and type <
Hold down the Alt-Key and type 0179
Hold down the Alt-Key and type 0163
Note: Per KDEP letter dated March 17, 2006. When entering data in the DMR form, if there is a
greater than sign (>) in the results, enter the number without the sign. A note must be made that
the number had a greater than sign and as to why there was a greater than sign. No changes are
needed when other signs are used. This is for the monthly and weekly average data.
53
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LFUCG Laboratory
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Revision Number 1
Last Revised 09/09
m-ColiBlue24 Method for the Determination of Total Coliforms and E. coli
HACH Method 10029
Federal Register (Dec. 1, 1999, FR Vol. 64, No. 230, 67449-67467)
40 CFR 141
1.
2.
3.
Scope, Significance to Process and Application
1.1
The coliform analysis is applicable to investigations of stream pollution, raw
water sources, and wastewater treatment systems.
1.2
m-ColiBlue24 broth simultaneously detects and identifies both total coliforms and
Escherichia coli (E. coli).
Summary of Method
2.1
Executive Summary
The sample is filtered through a 0.45 μm Millipore® membrane filter. The filter is
placed on a Pall® filter pad containing media in a sterile Petri dish. The samples
are then incubated at 35.0°C ± 0.5°C for 24 hours ± 2 hours. Colonies are
counted, blue colonies are enumerated as E. coli and other coliforms are red; total
coliforms are the sum of the two.
2.2
Discussion
m-ColiBlue24 is a single-step MF procedure that incorporates specific noncoliform growth inhibitors and a selective enzymatic indicator to allow for
simultaneous detection and quantitation of both E. coli and total coliforms. Fecal
coliforms are defined as. The major species of gram-negative, non-spore forming
rods is Escherichia coli, which indicates fecal pollution and the presence of
enteric pathogens. m-ColiBlue24 method has been approved by the U.S. EPA for
monitoring drinking water and wastewater using a 24-hour incubation period. It
can also be used to detect coliforms in other types of water (i.e. surface, ground,
well, recreational).
Health & Safety Precautions
3.1
3.2
3.3
3.4
All municipal and industrial wastewaters are potentially hazardous. Gloves and
safety glasses should be worn when dispensing these samples.
Possible exposure to enteric pathogens. Care must be taken to avoid undue
exposure.
A flame is used to sterilize forceps. Maintain the area around the flame clear.
Contaminated (used) Petri dishes and lab equipment must be placed in
Biohazardous waste container. This Biohazardous waste container is autoclaved
before disposal.
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4.
Sample Handling and Preservation
4.1
4.2
4.3
5.
Reagents
5.1
5.2
5.3
5.4
5.5
6.
Peptone powder
Peptone buffer solution pH 7.00 ± 0.1 (stored at 4°C)
m-ColiBlue24 media in plastic ampule (stored at 4°C)
Sodium thiosulfate 10% solution
Lysol disinfectant, 20% solution
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9
6.10
6.11
6.12
6.13
6.14
6.15
6.16
6.17
6.18
7.
Samples should be collected in clean, sterile glass or plastic containers.
If chlorine is in the sample, containers should be treated with 4 drops of
10% Sodium thiosulfate before autoclaving.
Run test immediately after sampling, or preserve sample at 4°C for a maximum of
6 hours.
Vacuum flask
Pall® Life Sciences single use 50 mm Petri dishes with pads
Millipore® sterile 0.45 μm 47 mm filter
Forceps
4.5 X 9 inch sterile sampling bags
Bunsen burner and striker
Pipettes and sterile tips
Sterilized filter holder (plastic or glass)
Gable topped water bath at 35.0°C ± 0.5°C
Thermometer
ASTM Thermometer
Tower Steam Indicator Strips
ODO-Clave® Heat Activated Deodorant Pads
Autoclavable Biohazard waste bags and deposit box
Autoclave
Sterile blue sheets
Indicator tape
Autoclavable Nalgene® squeeze bottles for peptone
Interferences
7.1
7.2
7.3
Bacteria from the surrounding environment.
Cross contamination from one sample to the next.
Lack of aseptic techniques.
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8.
Procedures
8.1
Steps
1) Clean work area with Lysol disinfectant, 20% solution.
2)
Light Bunsen burner with striker.
3)
Open sterile filter holder. Use sterile blue sheet as a sterile field. Indicator
trip and tape should indicate that the filter has been sterilized.
4)
Use Pall® petri dishes with sterile pad already in dish.
5)
Break open m-ColiBlue ampule of media and pour the contents evenly over
the absorbent pad and place the lid on the Petri dish.
6)
Decant excess media and cover dish to protect sterile pad.
7)
Place the bottom of the sterile filter holder onto the vacuum flask.
8)
Flame forceps, remove sterilized filter from packaging and place onto
sterilized filter holder (grid side up). Do not touch the filter with anything
except the forceps.
9)
Place or clamp the top unit onto filter holder.
10) Gently mix sample.
11) In advance, determine sample volume that will yield 20-60 fecal coliform
units (FCU).
12) If the volume of sample to be used is 0.1 to 5 mL, pour approximately 10
mL of peptone into filter unit before dispensing sample (Turn on vacuum
after the sample is introduced).
13) For sample volumes 5 to 50 mL, use sterile pipettes for dispensing into filter
unit.
14) Do not touch the inside of the filter holder unit. Do not allow the pipette tip
to touch the filter.
15) For sample volumes 50 to 100 mL, pour sample into cylinder and use the
scale on the side of the cylinder for measurement.
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16) Turn on vacuum. Once sample has filtered through, turn off vacuum.
17) Rinse top of the filter unit with peptone two times using autoclaved peptone
in a Nalgene® squeeze bottle.
18) Turn on vacuum to drain peptone. Turn off vacuum.
19) Flame forceps.
20) Remove top of the filter unit and place on the sterile blue field.
21) Open Petri dish.
22) Use sterile (flamed) forceps to grab the edge of the filter and remove it from
the filter holder unit.
23) Place filter, grid side up, onto edge of the Petri dish and gently slide it onto
the surface of the media saturated pad. Check for trapped air under the filter
and make sure the filter touches the entire pad.
24) Replace the Petri dish lid.
25) Place Petri dishes into a 4.5 X 9 inch sterile sampling bag. Make sure not
contaminate the inside of the bag. Seal bag.
26) Place bag, with Petri dishes face down, into water bath at 35.0°C ± 0.5°C for
24 ± 2 hours.
27) Log initials, time, and date in the Microbiology bench sheet.
28) After 24 ± 2 hours, count the colonies. Use a 10 to 15X stereoscopic
microscope, if necessary. If no red or blue colonies appear, the sample can
be considered negative. If red or blue colonies appear at 24 hours
incubation, the result is positive for total coliforms or E. coli, respectively.
No confirmation step is required.
29) Log results, initials, time, and date in the Microbiology bench sheet.
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8.2
Before and After Blanks
1)
2)
3)
4)
5)
6)
7)
8)
9)
10)
11)
8.3
Blanks must be analyzed before and after filtration of a set of samples.
Before any sample is filtered, place a sterile filter in the filter holder unit.
Add 50 mL peptone buffer to filter unit.
Turn on vacuum and filter the buffer, this will be the Before Blank.
Remove and plate filter as indicated in Section 8.1.
Proceed with sample filtration and plating.
Place a sterile filter in the filter holder unit.
Add 50 mL peptone buffer to filter unit.
Turn on vacuum and filter the buffer, this will be the After Blank.
Remove and plate filter as indicated in Section 8.1.
Log results, initials, time, and date in the Microbiology bench sheet.
Peptone Buffer Preparation
See SOP-Fecal Coliform, Section 8.4 for peptone buffer preparation.
8.4
9.
Helpful Hints
1)
If the bacterial density of the sample is unknown, filter and plate out several
volumes or dilutions in order to achieve a countable density. The volumes
and/or dilutions should be expected to yield a countable membrane. In
addition, select two additional quantities representing one-tenth and ten
times this volume, respectively.
2)
Separate filter holder units may be required during a set of samples. These
will be indicated in the bench sheet.
3)
Do not use damaged or bent membrane filters.
4)
Rinse the filter unit thoroughly with Peptone buffer to avoid cross
contamination.
QA/QC Requirements
9.1
9.2
Before and After Blanks must be run with each set of samples tested.
One duplicate per test series must be run.
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10.
Expected Results
10.1
11.
Data Analysis and Calculations
11.1
11.2
12.
KPDES Permit Requirements
Currently there are no permit requirement on E.coli.
See SOP – Fecal Coliform.
Blue colonies are counted as E. coli and red colonies are other coliforms; total
coliforms are the sum of the two.
Bibliography
12.1
HACH Method 10029, Coliforms: Membrane Filtration. 2009.
12.2
U.S. EPA. Microbiological Methods for Monitoring the Environment: Water and
Wastes. EPA 600/8-78-017. U.S. Environmental Protection Agency;
Environmental Monitoring and Support Laboratory, Office of Research and
Development, Washington, DC, 1978. Page 124.
12.3
Federal Register. Dec. 1, 1999, FR Vol. 64, No. 230, 67449-67467.
12.4
Code of Federal Regulations (CFR). 40 CFR 141. National primary drinking
water regulations. U.S. Environmental Protection Agency.
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LFUCG Laboratory
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Last Revised 09/09
Hardness, Total (mg/L as CaCO3; Titrimetric, EDTA)
HACH Method 8213 Hardness, Total with a Digital Titrator
Standard Methods 2340-C, U.S. EPA Method 130.2
1.
Scope, Significance to Process and Application
1.1
2.
3.
Summary of Method
2.1
Executive Summary
The EDTA titration method measures the calcium and magnesium ions and may
be applied with appropriate modification to any kind of water. The procedure
affords a means of rapid analysis.
2.2
Discussion
Sample is titrated with 0.800 M EDTA titrant until the color changes from red to
pure blue. The EDTA complexes calcium and magnesium ions. Color change
occurs when all calcium and magnesium ions are complexed, indicating the end
point of titration.
Health & Safety Precautions
3.1
3.2
3.3
4.
Hardness is defined as a characteristic of water which represents the total
concentration of calcium and magnesium expressed as their calcium carbonate
equivalent.
Watch out for broken glass from cylinders and porcelain dishes.
Wastewater samples have the potential to be hazardous, use appropriate caution.
Use of Nitric Acid, use appropriate caution.
Sample Handling and Preservation
4.1
4.2
4.3
4.4
Collect samples in plastic or glass containers that have been washed with a
detergent and rinsed with tap water, 1:1 Nitric Acid Solution, and Nanopure
water.
Analyze immediately or perform preservation.
To preserve the sample, add 1.5 mL of Nitric Acid per liter of sample. Check the
sample to assure that the pH is 2 or less.
Store samples at 4oC or below.
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5.
Reagents
5.1
5.2
5.3
6.
Equipment & Lab Ware
6.1
6.2
6.3
6.4
7.
HACH Digital burette.
100 mL graduated cylinder.
Porcelain dish.
Stirrer and magnet.
Interferences
7.1
7.2
8.
Hardness 1 Buffer Solution.
ManVer 2 Hardness Indicator Powder Pillow.
0.800 M EDTA Titration Cartridge.
Some metal ions interfere by causing fading or indistinct endpoint or by
stoichiometric consumption of EDTA.
Suspended or colloidal organic matter also may interfere with the endpoint.
Procedures
8.1
Steps
1) Attach a clean, horse-shoe bent delivery tube to a 0.800 M EDTA Titration
Cartridge connected to the HACH digital burette.
2)
Flush the delivery tube by turning the delivery knob on the burette to eject a
few drops of titrant. Reset the counter to zero and wipe tip.
3)
Measure 100 mL of sample with a clean 100-mL graduated cylinder.
4)
Pour the sample into a clean porcelain dish.
5)
Add 2 mL of Buffer Solution, Hardness 1, and stir.
6)
Add the contents of one ManVer 2 Hardness Indicator Powder Pillow and
stir.
7)
Titrate the sample with 0.800 M EDTA titrant until the color changes from
red to pure blue.
8)
Read the concentration of total hardness (as mg/L CaCO3) directly from the
digital counter window.
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9.
QA/QC Requirements
9.1
10.
None required.
Expected Results
10.1
KPDES Permit Requirements
None required. Monitored quarterly by 24-hour composite sample, concurrent
with whole effluent testing.
10.2
Process Ranges 1
Average Annual Hardness = 247 mg/L
Minimum monthly average = 177 mg/L
Maximum monthly average = 321 mg/L
1
11.
Data Analysis and Calculations
11.1
12.
Based on DMR data (January 1, 2001 through June 30, 2006).
None required.
Bibliography
12.1
HACH Water Analysis Handbook. Method 8213 Hardness, Total with a Digital
Titrator. 2nd Edition. 1992. HACH Company, Loveland, CO.
12.2
Standard Methods 2340-C. APHA-American Public Health Association Standard
Methods for the Examination of Water and Wastewater; 21th edition ed.;
American Water Works Association and Water Pollution Control Federation:
Washington, DC, 2005.
12.3
U.S. EPA. Method 130.2. Methods for Chemical Analysis of Water and Wastes.
EPA-600-4-79-020. U.S. Environmental Protection Agency; Office of Research
and Development, Washington, DC, 1982.
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Last Revised 09/09
pH (Electrometric)
EPA Method 150.1 pH. Issued 1971 (Editorial revision 1978 and 1982)
1.
Scope, Significance to Process and Application
1.1
1.2
2.
3.
Summary of Method
2.1
Executive Summary
The pH of a sample is determined electrometrically using either a glass electrode
in combination with a reference potential or a combination electrode.
2.2
Discussion
There is no color change during analysis. pH stands for power of hydrogen, a
measure of hydrogen ion concentration in solution.
Health & Safety Precautions
3.1
3.2
4.
Watch out for broken glass from beakers and cylinders.
Wastewater samples should be considered potentially hazardous. Use proper
personal protective equipment.
Sample Handling and Preservation
4.1
4.2
5.
At a given temperature the intensity of the acidic or basic character of a solution
is indicated by a pH or hydrogen ion activity.
A pH meter is accurate and reproducible to 0.1 pH unit with a range of 0 to 14
and equipped with a temperature compensation adjustment.
Samples should be analyzed as soon as possible within a 15 minute time window.
Sample containers should be filled completely and kept sealed prior to analysis.
Reagents
5.1
5.2
5.3
5.4
5.5
5.6
Buffer Solution pH – 4.00 (color coded red)
Buffer Solution pH – 7.00 (color coded yellow)
Buffer Solution pH – 10.00 (color coded blue)
Buffer Solution pH – 6.86
Nanopure Grade Laboratory Water
Reference Electrode Filling Solution
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6.
Equipment & Lab Ware
6.1
6.2
7.
Interferences
7.1
7.2
8.
Fisher Scientific AR50 pH Meter.
Stirrer probe.
pH measurements are affected by temperature and can cause the reading to drift.
Grease and other debris on the probe can cause inaccurate readings.
Procedures
8.1
Calibration
1)
Before calibrating, ensure the hole on the side of the pH probe is open.
2)
Weekly - Replace Reference Filling Solution.
a. Push down on probe to release filling solution.
b. Insert Reference Filling Solution bottle tip into hole of probe and
flush with solution.
c. Refill probe with Reference Electrode Filling Solution until inner
workings of probe are covered.
3)
Touch the meter screen (anywhere), until you hear a beep.
4)
Touch the “pH button” and wait until you hear a beep.
5)
Touch the “std button” on the upper right corner of the screen.
6)
Place probes into a beaker of Buffer Solution pH – 7.00 (yellow color
coded). Turn on stirrer.
7)
Touch the “clear button” to remove previous standardization.
8)
Write down the temperature of the buffer and determine what the pH for the
buffer will be at this temperature from chart.
Temperature oC
10
15
20
25
30
Buffer Solution pH – 7.00 (yellow color coded)
7.07
7.05
7.03
7.00
6.99
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9)
Type the value for the pH 7.0 buffer, at the given temperature, then press
“enter” key.
10) Turn off stirrer and rinse probes with Nanopure water.
11) Place probes into a beaker of Buffer Solution pH – 4.00 (red color coded).
Turn on stirrer.
12) Write down the temperature of the buffer and check what the pH for the
buffer will be at this temperature from chart.
Temperature oC
10
15
20
25
30
Buffer Solution pH – 4.00 (red color coded)
4.00
3.99
4.00
4.00
4.00
13) Touch the “std button” on the upper right corner of the screen.
14) Type the value for the pH 4.0 buffer, at the given temperature, then press the
“enter” key.
15) Turn off stirrer and rinse probes with Nanopure water.
16) Place probes into a beaker of Buffer Solution pH – 10.00 (blue color coded).
Turn on stirrer.
17) Write down the temperature of the buffer and determine what the pH for the
buffer will be at this temperature from chart.
Temperature oC
10
15
20
25
30
Buffer Solution pH – 10.00 (blue color coded)
10.19
10.12
10.06
10.00
9.94
18) Touch the “std button” on the upper right corner of the screen.
19) Type the value for the pH 10 buffer, at the given temperature, then press the
“enter” key.
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20) Turn off stirrer and rinse probes with Nanopure water.
21) Touch the “meas” button on the upper right side of the screen. Meter is now
calibrated and ready for sample measurements.
22) Record the slope, temperature, buffer values, time calibrated and initials in
the bench sheet and calibration folder.
8.2
8.3
8.4
Sample Measurements
1)
Touch the “meas” button on the upper right side of the screen.
2)
When testing samples for pH, place probe into sample container and record
the reading once the measurement has become stable. (“STABLE” will
appear once the meter recognizes that the measurement is stable).
3)
Turn off stirrer and rinse probes with Nanopure water between samples.
4)
When all analytes have been tested for that day, touch the “mode button” to
put meter into power save mode.
5)
Replace plastic sleeve or cap to cover hole in pH probe for safe storage.
6)
Store pH probe in pH 7.0 buffer solution.
Weekend Check Standard
1)
For Sunday operators, calibrate meter as described in Section 8.1.
2)
Measure pH of Buffer Solution pH – 6.86 to ensure meter is working
correctly. Record value of Check Standard in bench sheet.
3)
Proceed to measure samples as described in Section 8.2.
Helpful Hints
1) Avoid strong acids and greasy samples.
2) Make sure that probe is thoroughly rinsed between samples to avoid crosscontamination.
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9.
10.
QA/QC Requirements
9.1
If sample is not collected properly or analyzed within 15 minutes, another sample
must be obtained and analyzed.
9.2
A Check Standard (pH 6.86) must be analyzed on Sunday.
Expected Results
10.1
KPDES Plant Effluent Permit Requirements
1) 6.0 mg/L is the lowest pH allowed.
2) 9.0 mg/L is the highest pH allowed.
In the event that analysis results indicate values greater then KPDES permit
requirements, retest. If the value indicated by the retest is greater than KPDES permit
requirements, Immediately notify the Plant Superintendent and the Laboratory
Supervisor.
.
10.2 Process Ranges
Typical values for each plant are:
Town Branch Influent
Town Branch Effluent
11.
West Hickman Influent
West Hickman Effluent
7.1 – 7.4
6.4 – 7.7
Blue Sky Influent
Blue Sky Effluent
7.1 – 7.5
6.0 – 7.3
Data Analysis and Calculations
11.1
12.
7.3 – 7.4
6.7 – 8.9
None required.
Bibliography
12.1
U.S. EPA. Method 150.1 pH (Electrometric) Issued 1971 (Editorial revision 1978
and 1982). Methods for Chemical Analysis of Water and Wastes. EPA-600-4-79020; U.S. Environmental Protection Agency; Office of Research and
Development, Washington, DC, 1982.
12.2
AR50 Fisher Scientific User Manual. Fisher Scientific
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12.3
Code of Federal Regulations (CFR). 2003. Guidelines Establishing Test
Procedures for the Analysis of Pollutants. 40 CFR 136.3, Title 40, Chapter 1. U.S.
Environmental Protection Agency; U.S. Environmental Protection Agency. pg 5337.
TEMPERATURE to pH CHART
This chart is for color-coded buffers
only
Temperature
of the Buffer,
ºC
pH
pH
pH
Buffer Buffer Buffer
4.00
7.00
10.00
0
5
10
15
4.01
3.99
4.00
3.99
7.12
7.11
7.07
7.05
10.34
10.26
10.19
10.12
20
4.00
7.03
10.06
25
30
35
40
50
60
4.00
4.01
4.02
4.03
4.06
4.09
7.00
6.99
6.99
6.97
6.97
6.98
10.00
9.94
9.90
9.85
9.78
9.70
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Total Phosphorus Analysis
Ascorbic Acid Method
Phosphorus, Reactive (Orthophosphate), HACH Method 10209
Total Phosphorus, HACH Method 10210
TNT+ 843 LR (0.15 to 4.50 mg/L PO43– or 0.05 to 1.50 mg/L PO4–P)
TNT+ 844 HR (1.5 to 15.0 mg/L PO43– or 0.5 to 5.0 mg/L PO4–P)
TNT+ 845 UHR (6 to 60 mg/L PO43– or 2 to 20 mg/L PO4–P)
Reactive Phosphorus, EPA Method 365.1
Total Phosphorus, EPA Method 365.3
1.
Scope, Significance to Process and Application
1.1
2.
Phosphorus in wastewater is almost always present in the form of phosphates.
There are three major classes of phosphates; (1) Orthophosphates (fertilizer is
major source), (2) Polyphosphates (detergents and cleaning agents are major
sources), and (3) Organic Phosphates (biological waste is major source). Organic
Phosphates are also formed from orthophosphates during biological treatment of
waste streams. Analysis of Total Phosphorus includes all of the aforementioned
forms of Phosphorus. The reduction of Total Phosphorus levels throughout the
wastewater treatment process is highly important, as phosphorus concentrations in
plant effluents must be low enough (See Sec.10.1, Permit Requirements) to avoid
detrimental effects on the receiving environment, such as algae blooms.
Summary of Method
2.1
Executive Summary
Total phosphorus analysis at Town Branch Laboratory refers to the
spectrophotometric analysis of all phosphorus forms in a water/wastewater
sample that has been digested.
2.2
Discussion
Phosphates present in organic and condensed inorganic forms (meta-, pyro- or
other polyphosphates) are first converted to reactive orthophosphate in the Total
Phosphorus procedure. Treatment of the sample with acid and heat provides the
conditions for hydrolysis of the condensed inorganic forms. Organic phosphates
are also converted to orthophosphates in the Total Phosphorus procedure by
heating with acid and persulfate. The Reactive Phosphorus procedure measures
only the reactive (ortho) phosphorus present in the sample. The reactive or
orthophosphate ions react with molybdate and antimony ions in an acidic solution
to form an antimonyl phosphomolybdate complex, which is reduced by ascorbic
acid to phosphomolybdenum blue. Test results are measured at 890 nm with a
HACH DR5000 spectrophotometer.
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3.
Health & Safety Precautions
3.1
3.2
3.3
4.
Sample Handling and Preservation
4.1
4.2
4.3
4.4
4.5
4.6
5.
Collect samples in plastic or glass bottles that have been acid cleaned with 1:1
Hydrochloric acid solution and rinsed with Nanopure water.
Do not use commercial detergents containing phosphate for cleaning glassware
used in this test.
Analyze samples immediately after collection for best results.
If prompt analysis is impossible, preserve samples for Total Phosphorus up to 28
days by adjusting the pH to 2 or less with concentrated Sulfuric acid (about 2 mL
per liter) and storing at 4 °C.
Samples to be analyzed for Reactive Phosphorus should not be preserved with
acid: store Reactive Phosphorus samples at 4 °C and analyze within 48 hours.
Warm stored samples to 15–25 °C and neutralize with 5.0 N NaOH before
analysis if acid has been added.
Reagents
5.1
5.2
5.3
5.4
5.5
6.
During sample digestion, the digester block is HOT (100˚C), Burn Hazard. Use
digester block safety shields. In addition, the capped glass sample vials contain
hot (100˚C) acidic solution under pressure.
Watch out for broken glass from beakers and cylinders.
Wastewater samples should be considered potentially hazardous. Use proper
personal protective equipment.
Phosphorus, Reactive and Total, TNTplus LR Reagent Set (HACH TNT843)
Phosphorus, Reactive and Total, TNTplus HR Reagent Set (HACH TNT844)
Phosphorus, Reactive and Total, TNTplus UHR Reagent Set (HACH TNT845)
Nanopure Water
Phosphate Standard Solution 100 mg/L as PO43-
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
6.6
6.7
HACH DRB200 Reactor with Test’N Tube block and safety shields
TNTplus reactor adapter sleeves (16-mm to 13-mm diameter)
HACH DR5000 Spectrophotometer
Adjustable volume pipettes (100-1000 µL) with tips
Adjustable volume pipettes (1000-5000 µL) with tips
Test Tube Rack
Vials with 9 and 19 mL of Nanopure water for dilutions
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7.
Interferences
7.1
7.2
8.
Do not use commercial detergents containing phosphate for cleaning any lab ware
utilized in this method.
Excess Turbidity.
Procedures
8.1
TNTplus 843 Total Phosphorous LR (0.05–1.50 mg/L PO4–P)
1)
2)
3)
4)
5)
6)
7)
8)
9)
10)
11)
12)
13)
8.2
Turn on the DRB200 Reactor. Heat to 100 °C. For DRB200 Reactors with
16-mm wells, make sure the 16- to 13-mm adapter sleeves are in each well
before turning on the reactor.
Carefully remove the protective foil lid from the DosiCap™ Zip. Unscrew
the cap from the vial.
Carefully pipet 2.0 mL of sample into the vial.
Flip the DosiCap Zip over so the reagent side faces the vial. Screw the cap
tightly onto the vial.
Shake the capped vial with 2–3 times to dissolve the reagent in the cap.
Verify that the reagent has dissolved by looking down through the open end
of the DosiCap Zip.
Insert the vial in the DRB200 Reactor. Close the protective cover. Heat for 1
hour at 100 °C.
After the timer expires, carefully remove the hot vial from the reactor. Insert
it in a test tube rack and allow to cool to room temperature (15–25 °C).
Pipet 0.2 mL (200 μL) of Reagent B into the cooled vial. Immediately close
the Reagent B container.
Screw a grey DosiCap C onto the vial.
Invert the capped vial 2–3 times to dissolve the reagent in the DosiCap.
Wait 10 minutes.
When the timer expires, invert the vial again 2–3 times. Clean the vial with
a Kim-Wipe and insert it into the DR5000 cell holder. The instrument reads
the barcode, then selects and performs the correct test. Results are in mg/L
PO4. No instrument Zero is required.
Note and record the indicated values on the bench sheet
TNTplus 843 Reactive Phosphorous LR (0.15–4.50 mg/L PO43–)
1)
2)
3)
4)
Carefully pipet 2.0 mL of sample into the vial.
Pipet 0.2 mL (200 μL) of Reagent B into the vial. Immediately close the
Reagent B container.
Screw a grey DosiCap C onto the vial.
Invert the capped vial 2–3 times to dissolve the reagent in the DosiCap.
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5)
6)
7)
8)
8.3
TNTplus 844 Total Phosphorous LR HR (0.5-5.0 mg/L PO4–P)
1)
2)
3)
4)
5)
6)
7)
8)
9)
10)
11)
12)
13)
14)
8.4
Wait 10 minutes.
When the timer expires, invert the vial again 2–3 times.
Clean the outside of the vial with a Kim-Wipe and insert it into the DR5000
cell holder. The instrument reads the barcode, then selects and performs the
correct test. Results are in mg/L PO4. No instrument Zero is required.
Note and record the indicated values on the bench sheet
Turn on the DRB200 Reactor. Heat to 100 °C. For DRB200 Reactors with
16-mm wells, make sure the 16- to 13-mm adapter sleeve are in each well
before turning on the reactor.
Carefully remove the protective foil lid from the DosiCap™ Zip. Unscrew
the cap from the vial.
Carefully pipet 0.5 mL (500 μL) of sample into the vial.
Flip the DosiCap Zip over so the reagent side faces the vial. Screw the cap
tightly onto the vial.
Shake the capped vial 2–3 times to dissolve the reagent in the cap. Verify
that the reagent has dissolved by looking down through the open end of the
DosiCap Zip.
Insert the vial in the DRB200 Reactor. Close the protective cover. Heat for 1
hour at 100 °C.
After the timer expires, carefully remove the hot vial from the reactor. Insert
it in a test tube rack and allow to cool to room temperature (15–25 °C).
Pipet 0.2 mL (200 μL) of Reagent B into the cooled vial. Immediately close
the Reagent B container.
Screw a grey DosiCap C onto the vial.
Invert the capped vial 2–3 times to dissolve the reagent in the DosiCap.
Wait 10 minutes.
When the timer expires, invert the vial again 2–3 times.
Clean the outside of the vial with a Kim-Wipe and insert it into the DR5000
cell holder. The instrument reads the barcode, then selects and performs the
correct test. Results are in mg/L PO4. No instrument Zero is required.
Note and record the indicated values on the bench sheet
TNTplus 844 Reactive Phosphorous HR (1.5-15.0 mg/L PO43–)
1)
2)
3)
4)
Carefully pipet 0.5 mL of sample into the vial.
Pipet 0.2 mL (200 μL) of Reagent B into the vial. Immediately close the
Reagent B container.
Screw a grey DosiCap C onto the vial.
Invert the capped vial 2–3 times to dissolve the reagent in the DosiCap.
Install the Light Shield if applicable.
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5)
6)
7)
8.5
TNTplus 845 Total Phosphorous LR UHR (2-20 mg/L PO4–P)
1)
2)
3)
4)
5)
6)
7)
8)
9)
10)
11)
12)
13)
14)
8.6
When the timer expires, invert the vial again 2–3 times.
Clean the outside of the vial with a Kim-Wipe and insert it into the DR5000
cell holder. The instrument reads the barcode, then selects and performs the
correct test. Results are in mg/L PO4. No instrument Zero is required.
Note and record the indicated values on the bench sheet
Turn on the DRB200 Reactor. Heat to 100 °C. For DRB200 Reactors with
16-mm wells, make sure the 16- to 13-mm adapter sleeve are in each well
before turning on the reactor.
Carefully remove the protective foil lid from the DosiCap™ Zip. Unscrew
the cap from the vial.
Carefully pipet 0.4 mL (400 μL) of sample into the vial.
Flip the DosiCap Zip over so the reagent side faces the vial. Screw the cap
tightly onto the vial.
Shake the capped vial 2–3 times to dissolve the reagent in the cap. Verify
that the reagent has dissolved by looking down through the open end of the
DosiCap Zip.
Insert the vial in the DRB200 Reactor. Close the protective cover. Heat for 1
hour at 100 °C.
After the timer expires, carefully remove the hot vial from the reactor. Insert
it in a test tube rack and allow to cool to room temperature (15–25 °C).
Pipet 0.5 mL (500 μL) of Reagent B into the cooled vial. Immediately close
the Reagent B container.
Screw a grey DosiCap C onto the vial.
Invert the capped vial 2–3 times to dissolve the reagent in the DosiCap.
Wait 10 minutes.
When the timer expires, invert the vial again 2–3 times.
Clean the outside of the vial with a Kim-Wipe and insert it into the DR5000
cell holder. The instrument reads the barcode, then selects and performs the
correct test. Results are in mg/L PO4. No instrument Zero is required.
Note and record the indicated values on the bench sheet
TNTplus 845 Reactive Phosphorous UHR (6-60 mg/L PO43–)
1)
2)
3)
4)
5)
6)
Carefully pipet 0.4 mL of sample into the vial.
Pipet 0.5 mL (500 μL) of Reagent B into the vial. Immediately close the
Reagent B container.
Screw a grey DosiCap C onto the vial.
Invert the capped vial 2–3 times to dissolve the reagent in the DosiCap.
Wait 10 minutes.
When the timer expires, invert the vial again 2–3 times.
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7)
8)
8.7
Clean the outside of the vial with a Kim-Wipe and insert it into the DR5000
cell holder. The instrument reads the barcode, then selects and performs the
correct test. Results are in mg/L PO4. No instrument Zero is required.
Note and record the indicated values on the bench sheet
Reagent Blanks
A reagent blank can be measured and the value subtracted from the results of each
test performed using the same reagent lot number. Use Nanopure water in place of
sample and perform the Total Phosphorus, Method 10210 or the Reactive
Phosphorus, Method 10209 test.
To subtract the value of the blank from a series of measurements:
1. Measure the blank as in step 12 of the Total Phosphorus, Method 10210 test
or step 7 of the Reactive Phosphorus, Method 10209 test.
2. Activate the Reagent Blank feature. The measured value of the blank is shown
in the highlighted box.
3. Accept the value shown. The reagent blank value will be subtracted from all
results until the function is turned off or a different method is selected.
Alternately, the blank can be recorded and entered at any later time by pressing
the highlighted box and using the keypad to enter the value.
8.8
Sample Blanks
Color or turbidity in samples can cause high results. The digestion in the total
phosphate procedure usually destroys all color and turbidity and a sample blank is
not required.
To compensate for color or turbidity in the reactive phosphate procedure, the
color forming reagent that is present in the DosiCap C is not added.
To determine the sample blank for reactive phosphorus:
1. Run the Reactive Phosphorus, Method 10209 test, but do not add the DosiCap
C in step 3.
2. Cap the vial with the original DosiCap Zip but do not remove the foil. Use the
side of the cap without the reagent.
3. Subtract the value obtained in step 7 from the value obtained on the original
reactive phosphate sample to give the corrected sample concentration.
Alternatively, reactive phosphate samples that contain only turbidity may be first
filtered through a membrane filter and then analyzed. Samples without color or
turbidity do not require sample blanks.
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8.9
9.
Helpful Hints
1)
Analysis results are directly proportional to sample volumes; therefore it is
very important that accurate sample volume measurements are made.
Correct the test results for volume dilutions.
2)
The TNTplus test vials are cuvettes to be analyzed spectrophotometrically,
and must provide a clear optical path. Prior to reading, clean the vials by
wiping with a Kim-Wipe.
3)
When washing lab ware involved with this method, use only phosphate
free cleaning agents, 1:1 HCL solution is recommended, followed by
thorough Nanopure water rinse. Note: Most of the lab ware used in this
method is disposable.
Standard Preparation
A 100 mg/L as PO43- Phosphate Standard Solution is used as the stock to make standard
dilutions. The stock is located in the Room Temp Fridge.
ULR Low Standard (1.01 mg/L)
Combine 1.00 mL of the 10.1 mg/L High Standard with 9.0 mL Nanopure water.
HR High Standard (10.1 mg/L)
Phosphate Standard solution (100 mg/L) to 200 mL volumetric flask. Dilute to
200 mL with Nanopure water.
10.
QA/QC Requirements
10.1
A High Standard (10.1 mg/L) and a Low Standard (1.01 mg/L) must be run with
every analytical run.
10.2
A total of 5% of all samples must be run in duplicate.
10.3
Data acceptance criteria:
10.3.1 Analysis values for Standards must agree within 10% of the Standard’s
known value.
10.3.2 Duplicate values must agree within 5%.
If these criteria are not met, corrective action is indicated. See Quality Assurance
Program (QAP) Sec. 15 “Corrective Action Policies and Procedures”.
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11.
Expected Results
11.1
KPDES Permit Requirements
1)
Town Branch Wastewater Treatment Plant has no effluent limitations on
Total Phosphorus. Plant effluent composite samples are analyzed daily.
Monthly and weekly averages are reported.
2)
West Hickman Wastewater Treatment Plant effluent limitations are:
a. November 1st through April 30th - a monthly average of 1 mg/L and a
weekly average of 2 mg/L.
b. May 1st through October 31st - there are no effluent limitations on
Total Phosphorus.
Plant effluent composite samples are analyzed daily. Monthly and weekly
averages are reported.
In the event that analysis results indicate values greater than the KPDES permit
requirements, retest. If the value indicated by the retest is greater than KPDES
permit requirements, Immediately notify the Plant Superintendent and the
Laboratory Supervisor.
11.2
12.
Process Ranges
Typical phosphorus concentration values for plant influent vary primarily due to
rainfall. Within the treatment process phosphorus concentrations can vary due to
microbiological processes involving the release and uptake of phosphorus forms.
Typical values for each plant are:
Town Branch Influent
4 mg/L to 9 mg/L
Town Branch Effluent
2 mg/L to 4 mg/L
West Hickman Influent 2 mg/L to 12 mg/L
West Hickman Effluent
<1.0 mg/L
Data Analysis and Calculations
12.1
Concentration values are read directly from the DR5000 spectrophotometer.
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13.
Bibliography
12.1
HACH. DOC316.53.01124. Phosphorus, Reactive (Orthophosphate) and Total
Phosphorus. TNTplus 843. HACH Company, Loveland, CO, 2008.
12.2
HACH. DOC316.53.01125. Phosphorus, Reactive (Orthophosphate) and Total
Phosphorus. TNTplus 844. HACH Company, Loveland, CO, 2008.
12.3
HACH. DOC316.53.01126. Phosphorus, Reactive (Orthophosphate) and Total
Phosphorus. TNTplus 845. HACH Company, Loveland, CO, 2008.
12.4
HACH DR5000 Procedure Manual. HACH Company, Loveland, CO, 2008.
12.5
Standard Methods Part 4500-P. Phosphorous. APHA-American Public Health
Association Standard Methods for the Examination of Water and Wastewater;
21th edition ed.; American Water Works Association and Water Pollution Control
Federation: Washington, DC, 2005.
12.6
U.S. EPA Method 365.1 Phosphorous, All Forms (Colorimetric, Automated,
Ascorbic Acid). Methods for Chemical Analysis of Water and Wastes. EPA-6004-79-020. U.S. Environmental Protection Agency; Office of Research and
Development, Washington, DC, 1982.
12.7
U.S. EPA Method 365.3 Phosphorous, All Forms (Colorimetric, Ascorbic Acid,
Two Reagent). Methods for Chemical Analysis of Water and Wastes. EPA-600-479-020. U.S. Environmental Protection Agency; Office of Research and
Development, Washington, DC, 1982.
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Settleable Solids
EPA Method 160.5 Residue Settleable Matter
Standard Methods Part 2540-F
1.
Scope, Significance to Process and Application
1.1
1.2
2.
3.
Summary of Method
2.1
Executive Summary
Settleable solids is the term used for material settling out of suspension within a
defined period of time. It may include floating material, depending on the
technique. Solids analyses are important in the control of biological and physical
wastewater treatment processes and for assessing compliance with regulatory
agency effluent limitations. The settled volume of a biological suspension is
useful for routine activated sludge plant control.
2.2
Discussion
Settleable matter is measured volumetrically with an Imhoff Cone and a
Settlometer after 1 hour. A 1000 mL sample of mixed liquor is allowed to settle
and readings are taken after 30 minutes and at 1 hour intervals thereafter.
Health & Safety Precautions
3.1
4.
Wastewater samples should be considered potentially hazardous. Use proper
personal protective equipment.
Sample Handling and Preservation
4.1
4.2
4.3
4.4
4.5
5.
This method is applicable to surface and saline waters, domestic and industrial
wastes.
The practical lower limit of the determination is about 1 mL/L/hour.
Samples should be collected in plastic or glass containers.
No preservative is required.
Mixed liquor samples should be taken at effluent end of aeration tanks.
Care should be taken to minimize floc break-up during Settlometer analysis.
Maximum holding time is 7 days.
Reagents
5.1
None required.
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6.
Equipment & Lab Ware
6.1
6.2
6.3
7.
Interferences
7.1
8.
Imhoff Cone
Settlometer
Timer
None
Procedures
8.1
Imhoff Cone
1)
2)
3)
4)
5)
8.2
Mixed Liquor
1)
2)
3)
4)
5)
9.
Thoroughly mix samples and fill Imhoff Cones to the 1 Liter mark.
Set timer for Imhoff Cone samples for 45 minutes and additional 15
minutes.
After the Imhoff samples have settled for 45 minutes, stir samples gently to
remove suspended solids that may be on the surfaces of the Imhoff Cone.
After the Imhoff samples have settled for additional 15 minutes, record the
amount of settled solids in mL/L.
Record date of sample, date analysis completed, and initial in bench sheet.
Thoroughly mix sample (without aerating the sample) and fill Settlometer to
the 1 Liter mark.
Set timer for Settlometer sample for 30 minutes, 1 hr., 2 hr., 3 hr., and 4 hr.
After the Settlometer sample has settled for 30 minutes, record the amount
of settled solids in mL/L.
Observe mixed liquor throughout the 4 hour time span. If the blanket rises
before 4 hours, record the rise time in the bench sheet. If after 4 hours the
blanket has not risen, record >4 hours for rise time in bench sheet.
Record date of sample, date analysis completed and initial in bench sheet.
QA/QC Requirements
9.1
None Required
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10.
Expected Results
10.1
KPDES Permit Requirements
1)
No permit requirements.
10.2
Process Ranges
Typical values (mL/L) for each plant are:
Town Branch Influent
4.74 – 10.64
Town Branch Effluent
0.02 – 0.26
West Hickman Influent
West Hickman Effluent
11.
Data Analysis and Calculations
11.1
11.2
11.3
11.4
11.5
12.
8.65 – 20.61
0.02 – 0.25
Record as milliliters per liter.
Always read the top of the solids column.
If a separation of the settleable and floating materials occurs, do not estimate the
floating materials.
Detection limit: 0.01 mL/L.
% Settled Sludge = (mL of sludge in settled mixed liquor X 100)/1000
Bibliography
12.1
U.S. EPA Method 160.5 Residue, Settleable Matter. Methods for Chemical
Analysis of Water and Wastes. EPA-600-4-79-020. U.S. Environmental
Protection Agency; Office of Research and Development, Washington, DC, 1982.
12.2
Standard Methods Part 2540-F. Settleable Solids. APHA-American Public Health
Association Standard Methods for the Examination of Water and Wastewater;
21th edition ed.; American Water Works Association and Water Pollution Control
Federation: Washington, DC, 2005.
12.3
Code of Federal Regulations (CFR). Guidelines Establishing Test Procedures for
the Analysis of Pollutants. 40 CFR 136.3, Title 40, Chapter 1. U.S. Environmental
Protection Agency; U.S. Environmental Protection Agency. pg 5-337. 2003.
12.4
Simplified Laboratory Procedures for Wastewater Examination. Water Pollution
Control Federation, Third Edition. pg 17-18. 1985.
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Total Suspended Solids (TSS)
EPA Method 160.2 Residue, Non-Filterable & Total Suspended Solids
Standard Methods Part 2540-D
1.
Scope, Significance to Process and Application
1.1
1.2
2.
3.
Summary of Method
2.1
Executive Summary
Total solids are the material residue left in a vessel after evaporation of a sample
and subsequent oven drying at a defined temperature. Total suspended solids
(TSS) is the portion of total solids that is retained by filter. Solids analyses are
important in the control of biological and physical wastewater treatment processes
and for assessing compliance with regulatory agency effluent limitations.
2.2
Discussion
A well-mixed sample is filtered through a standard filter, and the residue retained
on the filter is dried to constant weight at 103oC to 105oC. The filtrate from this
method may be used for Residue, Total filterable.
Health & Safety Precautions
3.1
3.2
3.3
4.
Watch out for broken glass from crucibles, cylinders, and beakers.
Wastewater samples should be considered potentially hazardous. Use proper
personal protective equipment.
Crucibles can be hot (103oC to 105oC), use proper gloves when handling.
Sample Handling and Preservation
4.1
4.2
4.3
5.
This method is applicable to drinking, surface, and saline waters, domestic and
industrial wastes.
The practical range of the determination is 10 mg/L to 20,000 mg/L.
Collect samples in plastic or glass containers.
No preservative required.
Maximum holding time 7 days.
Reagents
5.1
5.2
Drierite 8 mesh.
Nanopure Grade water.
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6.
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
7.
Interferences
7.1
7.2
8.
Analytical Balance
ASTM Class 1 weight set
Environmental Express Pre-weighed filters for TSS (F92447MM)
Oven 103oC to 105oC
Beakers, stirrer, tongs
Vacuum pump and vacuum flask, 500 mL
Graduated cylinders
Desiccators
Too much residue on the filter will entrap water and may require prolonged
drying.
Too much residue on the filter may stop the movement of water through the filter.
Procedures
8.1
Steps
1)
Calibrate balance using ASTM Class 1 weight set. Typically 200 g, 100 g,
500 mg, and 100 mg.
2)
Environmental Express filters are pre-weighed. An ID number and weight of
filter are shown on the aluminum pan. Record the ID number and the filter
weight (This is W1).
3)
Assemble the filtering apparatus with pre-weighed filter and apply vacuum.
4)
Rinse filter with Nanopure water.
5)
Mix sample.
6)
Using a graduated cylinder, measure the required sample volume (V), then
gently and slowly pour into filter. Rinse side of filter unit with Nanopure
water.
7)
Placed filter and residue back in the original aluminum pan.
8)
Dry in oven (103oC to 105oC) until constant weight is achieved.
9)
Allow pans to cool in desiccator for 20 minutes before weighing.
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10) Weight of filter and residue is W2.
11) EPA requires filters to be placed back in the oven an reweighed three times.
8.2
Helpful Hints
1) Drying filters overnight produces the best results.
2) Filters stuck to the aluminum pan will produce incorrect results, re-filter
sample and reweigh with a new filter, if possible.
9.
QA/QC Requirements
9.1
9.2
10.
Balance must be calibrated using ASTM Class 1 weight set before analysis.
EPA requires filters with samples be placed back in the oven an reweighed three
times.
Expected Results
10.1
KPDES Plant Effluent Permit Requirements
1) 30 mg/L for Monthly Average
45 mg/L for Weekly Average
In the event that analysis results indicate values greater then KPDES permit
requirements, retest. If the value indicated by the retest is greater than KPDES
permit requirements, Immediately notify the Plant Superintendent and the
Laboratory Supervisor.
10.2
11.
Process Ranges
Typical values (mg/L) for each plant are:
Town Branch Influent
89 – 485
Town Branch Effluent
9 – 31
West Hickman Influent
West Hickman Effluent
137 – 513
2 – 72
Blue Sky Influent
Blue Sky Effluent
134 – 880
2 – 14
(30 mL sample)
(500-800 mL sample)
Data Analysis and Calculations
11.1
Non-filterable residue, mg/L = (W2 - W1)/V*1,000,000
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12.
Bibliography
12.1
U.S. EPA Method 160.2 Residue, Non-Filterable & Total Suspended Solids.
Issued 1971. Methods for Chemical Analysis of Water and Wastes. EPA-600-479-020. U.S. Environmental Protection Agency; Office of Research and
Development, Washington, DC, 1982.
12.2
Standard Methods Part 2540-D. Total Suspended Solids Dried at 103oC to 105oC.
APHA-American Public Health Association Standard Methods for the
Examination of Water and Wastewater; 21th edition ed.; American Water Works
Association and Water Pollution Control Federation: Washington, DC, 2005.
12.3
Code of Federal Regulations (CFR). Guidelines Establishing Test Procedures for
the Analysis of Pollutants. 40 CFR 136.3, Title 40, Chapter 1. U.S. Environmental
Protection Agency; U.S. Environmental Protection Agency. pg 5-337. 2003.
12.4
Simplified Laboratory Procedures for Wastewater Examination. Water Pollution
Control Federation, Third Edition. pg 23-24. 1985.
12.5
Environmental Express, Inc.. http://www.envexp.com/index.asp 2009.
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Total Solids
EPA Method 160.3 Residue, Total, (Gravimetric, Dried at 103 – 105 oC)
Standard Methods Part 2540-B, Total Solids Dried at 103 – 105oC
1.
Scope, Significance to Process and Application
1.1
1.2
2.
3.
Summary of Method
2.1
Executive Summary
Total solids are the material residue left in a vessel after evaporation of a sample
and subsequent oven drying at a defined temperature. Typically, total solids
include total suspended solids (TSS) and total dissolved solids (or the portion of
total solids that passes through a filter). Solids analyses are important in the
control of biological and physical wastewater treatment processes and for
assessing compliance with regulatory agency effluent limitations.
2.2
Discussion
A well mixed aliquot of sample is quantitatively transferred to a pre-weighed
evaporating dish, evaporated to dryness at 103oC to 105oC, and weighed to
determine total solids.
Health & Safety Precautions
3.1
3.2
3.3
4.
Watch out for broken glass from crucibles, cylinders, and beakers.
Wastewater samples should be considered potentially hazardous. Use proper
personal protective equipment.
Crucibles can be hot (103oC to 105oC), use proper gloves when handling.
Sample Handling and Preservation
4.1
4.2
4.3
5.
This method is applicable to drinking, surface and saline waters, domestic and
industrial wastes.
The practical range of the determination is from 10 mg/L to 20,000 mg/L.
Collect samples in plastic or glass containers.
No preservative required.
Maximum holding time 7 days at 4oC.
Reagents
5.1
5.2
Drierite 8 mesh
Nanopure Grade water
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6.
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
7.
Interferences
7.1
7.2
8.
Denver Instrument Analytical Balance
ASTM Class 1 weight set
Vycor dishes or flat-bottom crucibles
Oven 103oC to 105oC
Plastic 150 mL beakers
Stirrers
Tongs
Desiccators
Non-representative particles such as leaves, sticks, fish, and lumps of fecal matter
should be excluded from the sample.
Floating oil and grease, if present, should not be included in the sample.
Procedures
8.1
Steps
1)
Clean and dry dishes in oven overnight.
2)
Remove dishes from oven and put into desiccator for at least 20 minutes.
3)
Calibrate Analytical Balance using ASTM Class 1 weight set.
4)
Weigh dish. This is W1.
5)
Add approximately 50 mL of mixed sample into the dish.
6)
Weigh dish and sample. This is W2.
7)
Turn on fume hood.
8)
Leave samples in oven overnight.
9)
Remove dishes from oven and put into desiccator for at least 20 minutes.
10)
The next day, calibrate Analytical Balance using ASTM Class 1 weight
set.
11)
Weigh dish and dry sample. This is W3.
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8.2
Helpful Hints
1)
Do not allow dishes to sit out in open air before weighing.
2)
9.
QA/QC Requirements
9.1
10.
Record oven temperature, date, and time. Make sure to initial records.
None required
Expected Results
10.1
KPDES Permit Requirements
1)
None required.
10.2
Process Ranges
Typical values (% total solids, mg/L) for each plant are:
Town Branch
W.A.S. Thickener
3.68
Raw Sludge to Thickener 0.41 – 0.97
Digested Sludge
1.49 – 1.84
Belt Press Cakes
15.35 – 19.68
West Hickman
Return Activated Sludge 0.79 – 1.16
W.A.S. to Thickener
0.79 – 1.16
Belt Press Cakes
16.45 – 18.94
11.
Data Analysis and Calculations
11.1
Total Solids mg/L = (W3 – W1)/(W2 – W1)*1,000,000
11.2
% Total Solids mg/L = (W3 – W1)/(W2 – W1)*100
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12.
Bibliography
12.1
U.S. EPA Method 160.3 Residue, Total (Gravimetric, Dried at 103 – 105oC).
Issued 1971. Methods for Chemical Analysis of Water and Wastes. EPA-600-479-020. U.S. Environmental Protection Agency; Office of Research and
Development, Washington, DC, 1982.
12.2
Standard Methods Part 2540-B. Total Solids dried at 103oC to 105oC. APHAAmerican Public Health Association Standard Methods for the Examination of
Water and Wastewater; 21th edition ed.; American Water Works Association and
Water Pollution Control Federation: Washington, DC, 2005.
12.3
Code of Federal Regulations (CFR). Guidelines Establishing Test Procedures for
the Analysis of Pollutants. 40 CFR 136.3, Title 40, Chapter 1. U.S. Environmental
Protection Agency; U.S. Environmental Protection Agency. pg 5-337. 2003.
12.4
Simplified Laboratory Procedures for Wastewater Examination. Water Pollution
Control Federation, Third Edition. pg 25-27. 1985.
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LFUCG Laboratory
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Volatile Acids
Standard Methods Part 5560-C Distillation Method
HACH Method 8291, Buret Titration Method
1.
Scope, Significance to Process and Application
1.1
1.2
1.3
1.4
2.
3.
Summary of Method
2.1
Executive Summary
Volatile fatty acids are classified as water-soluble fatty acids (up to six carbon
atoms) that can be distilled at atmospheric pressure because of co-distillation with
water. A sample is acidified with sulfuric acid, distilled, and the distillate is
titrated to the phenolphthalein end point with sodium hydroxide. The volume of
titrant is proportional to the volatile acids concentration. Results are expressed as
mg/L as acetic acid.
2.2
Discussion
Acidity of water is its quantitative capacity to react with a strong base to a
designated pH. Acidity is a measure of an aggregate property of water and can be
interpreted in terms of specific substances only when the chemical composition of
the sample is known. In this case, 125 mL is distilled, brought to 95oC, and
titrated until a slight pinkish color is obtained.
Health & Safety Precautions
3.1
3.2
3.3
3.4
3.5
3.6
4.
This technique recovers acids containing up to six carbon atoms.
Fractional recovery of each acid increases with increasing molecular weight.
Calculations and reporting are on the basis of acetic acid.
The method is often applicable for control purposes.
Watch out for broken glass from cylinders and beakers.
Wastewater samples should be considered potentially hazardous. Use proper
personal protective equipment.
Distillation flasks can be HOT, use proper gloves when handling.
Mercury from broken thermometers can be a safety hazard.
Sulfuric Acid is used during this analysis. Wear gloves, goggles and lab coat.
ALWAYS ADD ACID TO WATER - NOT WATER TO ACID!!!
Sample Handling and Preservation
4.1
None required.
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5.
Reagents
5.1
5.2
5.3
5.4
5.5
6.
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9
6.10
6.11
6.12
6.13
6.14
6.15
6.16
7.
Heating Mantles
Hot plate
Centrifuge tubes
Boiling flask
Condenser, about 76 cm long
Distillation Assembly
Adapter tubes
Erlenmeyer flasks
Thermometers with stoppers
Titrator
Digital Buret
Plastic funnels
5 mL pipette and tips
Timer
Finger, vinyl, leather gloves
100 mL & 20 mL graduated cylinders
Interferences
7.1
8.
Sulfuric acid 1:1
0.1 N Sodium hydroxide
Phenolphthalein Indicator
Nanopure Water
Glacial acetic acid
Hydrogen sulfide and carbon dioxide are liberated during distillation and can be
titrated to give a positive error. Eliminate error by discarding the first 15 mL of
distillate and account for this in the recovery factor.
Procedures
8.1
Steps
1)
Turn on heating mantles.
2)
Make sure hot plate is on.
3)
Fill centrifuge tubes to obtain 125 mL of sample. Centrifuge samples for
15 minutes.
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4)
Gently swirl tubes to mix, and centrifuge for an additional 15 minutes.
5)
Decant 125 mL of sample and pour into the Boiling flask.
6)
Add 125 mL of Nanopure water to the Boiling flask. Swirl to mix.
7)
Add 5 mL of 1:1 Sulfuric Acid to the Boiling flask. Swirl to mix
8)
Connect flask to a condenser and adapter tube.
9)
Distill at a rate of 5 mL/min.
10)
Make sure tap water is running and cooling the condenser.
11)
Set timer for 5, 20, and 20 more minutes.
12)
Collect the first 15 mL of distillate in 20 mL graduated cylinder and
discard. This takes approximately 5 minutes.
13)
Collect 150 mL of distillate in an Erlenmeyer flask. This takes
approximately 20 minutes.
14)
While wearing leather gloves, disconnect Boiling flask from distillation
unit.
15)
Transfer the Erlenmeyer flask to the hot plate, insert thermometer, and
stopper.
16)
Heat to 90oC ± 5oC. This takes approximately 20 minutes.
17)
While wearing finger gloves, remove flask from hot plate.
18)
Add 10 drops of Phenolphthalein Indicator. Swirl to mix.
19)
Titrate, drop-wise, with 0.1 N Sodium hydroxide (NaOH) using the Brand
Digital Buret II until first persistent pink color is obtained.
20)
Record in bench sheet end point, date sample collected, date analysis
performed, time analysis started, and initials.
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LFUCG Laboratory
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8.2
Recovery Factor Determination
To determine the recovery factor (f):
1)
Dilute 1.9 mL of glacial acetic acid in 1 L Nanopure water (2000 mg/L
acetic acid solution).
2)
Add 50 mL of prepared acetic acid solution to 1 L volumetric flask and
bring to 1 L with Nanopure water (final acetic acid concentration = 100
mg/L).
3)
Measure 250 mL of diluted acetic acid solution and distill as shown in
Section 8.1.
4)
Titrate drop wise with 0.1 N Sodium Hydroxide (NaOH) using the Brand
Digital Buret II until first persistent pink color is obtained.
5)
Calculations:
a = (volume of NaOH required)(0.1N)(60000)/250 mL
Recovery factor (f) = a/b , where b=100 mg/L
8.3
9.
QA/QC Requirements
9.1
10.
11.
Helpful Hints
1)
If there is no color change, it means no Sulfuric acid was added.
None required.
Expected Results
10.1
KPDES Permit Requirements
1)
None required.
10.2
Process Ranges
1)
Town Branch Digesters ranges are 35 – 73 mg/L.
2)
If volatile acids ratio is high for plant operations, run analysis again.
Data Analysis and Calculations
11.1
mg volatile acids as acetic acid/L = (mL NaOH)(0.1N)(60,000)/ (mL sample)(f)
Where: f = recovery factor
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12.
Bibliography
12.1
Standard Methods Part 5560-C, Distillation Method. APHA-American Public
Health Association Standard Methods for the Examination of Water and
Wastewater; 21th edition ed.; American Water Works Association and Water
Pollution Control Federation: Washington, DC, 2005.
12.2
HACH Water Analysis Handbook. Method 8291, Volatile Acids, Buret Titration
Method. 2nd Edition. HACH Company, Loveland, CO, 1992.
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Volatile Solids
EPA Method 160.4 Residue, Volatile, (Gravimetric, Ignition at 550oC), Issued 1971
Standard Methods Part 2540-E, Fixed and Volatile Solids Ignited at 550oC
1.
Scope, Significance to Process and Application
1.1
2.
3.
Summary of Method
2.1
Executive Summary
Total solids are the material residue left in a vessel after evaporation of a sample
and subsequent oven drying at a defined temperature. The total solids are then
ignited to a constant weight at 550oC and the weight loss on ignition represents
the volatile solids. Volatile solids analyses are important because they offer a
rough approximation of the amount of organic matter present in the solid fraction
of wastewater, activated sludge, and industrial waste.
2.2
Discussion
Volatile solids are the weight lost due to ignition. The remaining solids represent
the fixed total.
Health & Safety Precautions
3.1
3.2
3.3
4.
Watch out for broken glass from Vycor dishes, cylinders, and beakers.
Wastewater samples should be considered potentially hazardous. Use proper
personal protective equipment.
Dishes are HOT (550oC), use proper gloves when handling. Watch out for hot
surfaces.
Sample Handling and Preservation
4.1
5.
This method is applicable to drinking, surface and saline waters, domestic and
industrial wastes.
Do immediately after Total Solids Method (See SOP - Total Solids).
Reagents
5.1
Drierite 8 mesh.
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6.
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
6.6
6.7
7.
Interferences
7.1
7.2
8.
Negative errors in the volatile solids may be produced by loss of volatile matter
during drying.
Determination of low concentrations of volatile solids in the presence of high
fixed solids concentrations may be subject to considerable error.
Procedures
8.1
8.2
9.
Denver Instrument Analytical Balance
ASTM Class 1 weight set
Vycor dishes or flat bottom crucibles
Tongs
Thermolyne 30400 Muffle Furnace (550oC)
Desiccator
Hot Plate (103oC to 105oC)
Steps
1)
After Total Solids Method is completed (See SOP - Total Solids) place
dishes in Muffle Furnace. Use tongs and gloves.
2)
Ignite tares at 550 ± 50oC to constant weight (approximately 1 hr).
3)
Transfer tares to hot plate for 15 minutes.
4)
Transfer tares to desiccator for at least 20 minutes. (Note: Make sure the
dishes are cool enough not to melt the desiccator).
5)
Weigh tare and ash and record weight as W4.
Helpful Hints
1)
Transfer tares to hot plate so it doesn’t melt desiccator.
2)
Radiant heat from furnace can burn, wear gloves and use tongs.
QA/QC Requirements
9.1
None required.
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10.
Expected Results
10.1
KPDES Permit Requirements
None required.
If value is inconsistent, DNS (Data Not Supportable) is typed into computer.
10.2
11.
Data Analysis and Calculations
11.1
11.2
12.
Process Ranges
The practical determination range is 10 mg/L to 20,000 mg/L.
Volatile Solids (mg/L) = (W3 – W4)/ (W2 – W1) * 1,000,000
% Volatile Solids = (W3 – W4)/ (W3 – W1) * 100
Bibliography
12.1
U.S. EPA Method 160.4 Residue, Volatile, (Gravimetric, Ignition at 550oC),
Issued 1971. Methods for Chemical Analysis of Water and Wastes. EPA-600-479-020. U.S. Environmental Protection Agency; Office of Research and
Development, Washington, DC, 1982.
12.2
Standard Methods Part 2540-E, Fixed and Volatile Solids Ignited at 550oC.
APHA-American Public Health Association Standard Methods for the
Examination of Water and Wastewater; 21th edition ed.; American Water Works
Association and Water Pollution Control Federation: Washington, DC, 2005.
96
WEST HICKMAN CREEK SOPs
1
SOP – WH FIELD D.O.
West Hickman WWTP Laboratory
Page 2 of 219
Revision Number 3
Last Revised 09/09
WH Dissolved Oxygen Field Analysis (D.O.)
HACH Method 10360 Luminescent Dissolved Oxygen Probe Method
Proposed EPA Method 360.3 (Luminescence) for the Measurement of
Dissolved Oxygen in Water and Wastewater
1.
2.
Scope, Significance to Process and Application
1.1
Dissolved Oxygen (D.O.) analysis measures the concentration of oxygen that is
dissolved in a water sample.
1.2
This method is recommended for samples containing intense color or turbidity
which interferes with the Winkler method.
1.3
This method is recommended for work in the field, as the equipment is portable,
allowing hold times to be minimized.
1.4
KPDES Permit Limits on Plant Effluent Required a minimum of 7.0 mg/L.
1.5
Dissolved Oxygen concentration levels are very important in both process and
plant effluents. In process, dissolved oxygen is required by various organisms and
the plant effluent dissolved oxygen levels must be conducive to the receiving
environment and within permit limits (See Section 10.1).
Summary of Method
2.1
Executive Summary
Dissolved Oxygen is measured directly by a HACH model HQ40d portable meter
and HACH model LDO101 rugged field dissolved oxygen probe. After the meter
indicates a stable reading the operator/analyst records the value.
2.2
Discussion
The HACH LDO system uses a sensor coated with a luminescent material. Blue
light from an LED is transmitted onto the sensor surface, exciting the luminescent
material, which then emits red light as it relaxes. The presence of DO in the
process shortens the time it takes for the red light to be emitted. By measuring the
time lapse between when the blue light was transmitted and the red light is
emitted, a correlation is made to the concentration of DO in the effluent or other
solution. Between measurements, a red LED is used as an internal reference. The
measurement range for the method is 0.02 mg/L to 20.0 mg/L. The Method
Detection Limit (MDL; 40 CFR 136, Appendix B) has been determined as 0.05
mg/L and the Minimum Level (ML; Reference 15.4) has been set at 0.20 mg/L.
2
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West Hickman WWTP Laboratory
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3.
Health & Safety Precautions
3.1
3.2
4.
Glassware involved: possible cut hazard.
All municipal and industrial wastewaters are potentially hazardous.
Gloves and goggles should be worn when dispensing these samples.
Sample Handling and Preservation
4.1
4.2
Field measurements are obtained directly, therefore, sample handling may not
apply.
If samples are collected for analysis at another location (i.e. Laboratory), the
following apply:
4.2.1
Sample must be collected in a glass bottle (preferably a BOD bottle with
stopper) filled to top, with no airspace.
4.2.2 Sample must be analyzed immediately (15 minutes maximum on permit
samples).
4.3
5.
Reagents
5.1
6.
Nanopure Lab Water
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
7.
There is no applicable preservative with this method.
HACH HQ40d portable multi-meter
HACH Model LDO101 Rugged Dissolved Oxygen Probe
BOD bottles with 300 ml capacity and tapered ground glass stoppers
Sensor Cap replacements (HACH part # 5838000)
Calibration bottle for “Water-saturated Air” calibration method
Interferences
7.1
7.2
7.3
Salinity (salinity correction available, See Section 8.4.3 of the Users Manual).
Reactive gas: chlorine and hydrogen sulfide.
Air bubbles in sample or on surface of probe tip.
3
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West Hickman WWTP Laboratory
Page 4 of 219
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Last Revised 09/09
8.
Procedures
8.1
Calibration and Start Up
It is suggested that at the Users Manual be initially consulted when following
these procedures.
1) Press the power button on the HQ40d and allow the unit to perform its startup
self check routine.
2) Clean by rinsing with Nanopure lab water, then blot dry the probes tip with a
Kim-Wipe. Inspect the probe tip for indications contamination or damage.
3) Take a 300 mL BOD bottle containing approximately one inch of lab water,
cap and shake, remove cap and replace it with the probe.
4) Press Calibrate (blue button), the meter will prompt you to “Dry the probe and
place in water saturated air & press “Read”. Press “Read”, the screen will
scroll from 0 to 100%, then indicate “Calibration Complete”. Record from
display screen both the temperature and the dissolved oxygen value (indicated
under the temperature). Log the values on the dissolved oxygen calibration
section of the Dissolved Oxygen bench sheet under D=Temperature and
E=Dissolved Oxygen from HQ40d.
5) Note the barometric pressure value from the laboratory barometer (located
adjacent to the D.O. meter) and record on the dissolved oxygen calibration
section of the Dissolved Oxygen bench sheet under “Barometer Reading”.
6) On the Lab computer, open the excel spreadsheet entitled “DO Meter
Calibration Sheet” and enter the barometric pressure, temperature and
dissolved oxygen values from the dissolved oxygen calibration section of the
Dissolved Oxygen bench sheet. The spreadsheet will calculate the “Dissolved
Oxygen Calibration Point”, the “Dissolved Oxygen @ 1 ATM” and the Slope
%. Transfer the three values onto the dissolved oxygen calibration section of
the Dissolved Oxygen bench sheet, then print a copy of the spreadsheet and
file it.
7) Note the difference between the Dissolved Oxygen from HQ40d and the
Dissolved Oxygen Calibration Point - if it is greater than 0.2 mg/L, then the
calibration is not acceptable and must be repeated until criteria is met.
4
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West Hickman WWTP Laboratory
Page 5 of 219
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8.2
Measurements
1) Make sure that the meter is properly calibrated.
2) Rinse the LDO101 probe tip with Nanopure lab water.
3) Place probe into a BOD bottle filled to the base of its neck with sample, assure
that there are no air bubbles on the surface of the probe tip. In the field, the
probe is lowered into the sample.
4) Press Read, screen will display “Stabilizing” and a progress bar will scroll
from 0 to 100%.
5) Reading stability is indicated by the appearance of a “Padlock” icon in the
upper left corner of the display screen. Record the indicated value, remove the
probe, and rinse tip with Nanopure water. Then proceed to the next sample or
store until needed.
6) DO NOT store probe in the BOD bottle containing water. Probe can be stored
dry on the bench top.
8.3
Helpful Hints
1) The meter is designed to be maintenance free, when needed, clean the exterior
with a damp cloth.
2) The probe’s maintenance consist of maintaining the probe tip clean, frequent
rinsing with Nanopure water is sufficient.
3) DO NOT scrub the sensor cap or lens.
4) DO NOT use any organic solvents on the sensor cap or probe body.
9.0
QA/QC Requirements
9.1
9.2
9.3
Meter must be calibrated a minimum of once per analysis day.
Permit sample hold times must be 15 minutes or less.
Probe condition must be properly maintained through routine cleaning (See
Section 8.3, Helpful Hints).
5
SOP – WH FIELD D.O.
West Hickman WWTP Laboratory
Page 6 of 219
Revision Number 3
Last Revised 09/09
10.
11.
Expected Results
10.1
KPDES Permit Requirements
7.0 mg/L is the lowest D.O. value allowable in a plant effluent sample at any
given time. In the event of an indicated value less than 7.0 mg/L, assure correct
calibration, resample, and retest. If the value indicated by retest is less than 7.0
mg/L, Immediately notify the Plant Superintendent and the Laboratory
Supervisor.
10.2
Process Ranges
Raw influent dissolved oxygen values are typically less than 1 mg/L.
Target values for mixed liquor dissolved oxygen concentration in the aeration
basins is 2.0 mg/L.
Plant effluent dissolved oxygen values must be maintained at or above 7.0 mg/L.
Data Analysis and Calculations
11.1
12.
None required, values are taken directly when measurement stability is indicated.
Bibliography
12.1
Report on the Validation of Proposed EPA Method 360.3 (Luminescence) for the
Measurement of Dissolved Oxygen in Water and Wastewater. August 2004.
HACH Company, Loveland, CO, 2004.
12.2
Memorandum: EPA Recommendation for the use of HACH method 10360
[Revision 1.1, January 2006] (ATP Case # N04-0013).
12.3
HACH HQ Series Portable Meter Users Manual, September 2006, Edition 5.
HACH Company, Loveland, CO, 2006.
6
SOP – WH pH
West Hickman WWTP Laboratory
Page 7 of 219
Revision Number 3
Last Revised 09/09
WH pH (Electrometric)
EPA Method 150.1 pH. Issued 1971 (Editorial revision 1978 and 1982)
HACH USEPA Electrode Method 8156
1.
Scope, Significance to Process and Application
1.1
1.2
2.
3.
Summary of Method
2.1
Executive Summary
The pH of a sample is determined electrometrically using either a glass electrode
in combination with a reference potential or a combination electrode.
2.2
Discussion
There is no color change during analysis. pH stands for power of hydrogen, a
measure of hydrogen ion concentration in solution.
Health & Safety Precautions
3.1
3.2
4.
Watch out for broken glass from beakers and cylinders.
Wastewater samples should be considered potentially hazardous. Use proper
personal protective equipment.
Sample Handling and Preservation
4.1
4.2
5.
At a given temperature the intensity of the acidic or basic character of a solution
is indicated by a pH or hydrogen ion activity.
A pH meter is accurate and reproducible to 0.1 pH unit with a range of 0 to 14
and equipped with a temperature compensation adjustment.
Samples should be analyzed as soon as possible within a 15 minute time window,
preferably in the field.
Collect samples in clean plastic or glass bottles. Sample containers should be
filled completely and kept sealed prior to analysis.
Reagents
5.1
5.2
5.3
5.4
5.5
Buffer Solution pH – 4.00 (color coded red)
Buffer Solution pH – 7.00 (color coded yellow)
Buffer Solution pH – 10.00 (color coded blue)
Nanopure Grade Laboratory Water
pH Electrode Storage Solution
7
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West Hickman WWTP Laboratory
Page 8 of 219
Revision Number 3
Last Revised 09/09
6.
Equipment & Lab Ware
6.1
6.2
7.
Interferences
7.1
7.2
8.
HQ40d Dual-Input Multi-Parameter Digital Meter
PHC101 IntelliCAL Rugged Gel Filled pH Electrode
pH measurements are affected by temperature and can cause the reading to drift.
Grease and other debris on the probe can cause inaccurate readings.
Procedures
8.1
Calibration
1)
Refer to the operation section of the electrode or meter manual to prepare
the PHC101 pH electrode and HQ40d meter.
2)
Turn the HQ40d meter on. Push the “Down” arrow.
3)
Make sure that the meter is set to measure to measure pH.
4)
In three separate beakers or appropriate containers, prepare fresh buffers of
pH 7.0, 4.0 and 10.0.
5)
Place probe in pH 7.0 buffer (yellow color coded).
6)
Turn on stir plate.
7)
Press READ and wait until a stable reading is obtained (lock icon).
8)
Rinse probe with Nanopure water.
9)
Place probe in pH 4.0 buffer (red color coded).
10) Turn on stir plate.
11) Press READ and wait until a stable reading is obtained (lock icon).
12) Rinse probe with Nanopure water.
13) Place probe in pH 10.0 buffer (blue color coded).
14) Turn on stir plate.
8
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West Hickman WWTP Laboratory
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Last Revised 09/09
15) Press READ and wait until a stable reading is obtained (lock icon).
16) Rinse probe with Nanopure water.
17) Press DONE and the meter will display all data gathered.
18) Make sure that the calibration slope is acceptable (typically -58 ±3 mV per
pH unit at 25°C).
19) Press STORE twice to accept calibration.
20) Rinse the electrode in Nanopure water and blot dry prior to sample
measurement. Rinse the electrode with Nanopure between measurements to
minimize contamination.
8.2
8.3
Sample Measurement
1)
Put the electrode in the sample. In the field, readings are taken directly.
2)
Turn on stir plate and press READ. For faster response, stir at a slow to
moderate rate.
3)
When the measurement is stable, store or record the pH and temperature
values. For the HQ40d meter, data is stored automatically when Press to
Read or Interval is selected in the Setup Measurement Mode. When
Continuous is selected, data will only be stored when the key under STORE
is pressed.
4)
Store the pH electrode in pH storage solution when not in use. See Sample
collection, preservation, general storage and cleaning for more details
Helpful Hints
1)
Avoid strong acids and greasy samples.
2)
Make sure that probe is thoroughly rinsed between samples so that cross
contamination does not occur.
3)
Storage of an electrode is based on how long the electrode will be stored,
how quickly the electrode needs to be used and the type of sample being
measured. For general storage, use the HACH storage solution or a 3M
Potassium chloride (KCl) solution.
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West Hickman WWTP Laboratory
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9.
10.
4)
A contaminated glass bulb or fouled electrode may cause slow response
times. Do not clean the bulb too often because the bulb life may shorten.
5)
To clean an electrode with general contamination, immerse the electrode tip
in 0.1N Hydrochloric acid (HCl). Then, immerse the electrode in 0.1N
Sodium hydroxide (NaOH) and again in 0.1N Hydrochloric acid, each for a
2-minute period. Rinse with Nanopure water and soak in Nanopure water
for at least 15 minutes.
6)
To clean an electrode contaminated with oils and fats, immerse the electrode
tip in a detergent solution. Use a soft brush or ultrasonic bath if necessary.
Avoid scratching the glass bulb.
QA/QC Requirements
9.1
If sample is not collected properly or analyzed within 15 minutes, another
sample must be obtained and analyzed.
9.2
Check electrode response
An electrode is responding properly if its calibration slope meets the slope
specifications of the electrode (typically -58 ±3 mV at 25°C).
9.3
Check calibration accuracy
Return the electrode to a calibration buffer and measure the pH to test the
system. Rinse and recondition the electrode before measuring subsequent
samples.
Expected Results
10.1
KPDES Permit Requirements
1) 6.0 mg/L is the lowest pH allowed.
2) 9.0 mg/L is the highest pH allowed.
In the event that analysis results indicate values greater then KPDES permit
requirements, retest. If the value indicated by the retest is greater than KPDES
permit requirements, Immediately notify the Plant Superintendent and the
Laboratory Supervisor.
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West Hickman WWTP Laboratory
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10.2
11.
7.3 – 7.4
6.7 – 8.9
West Hickman Influent
West Hickman Effluent
7.1 – 7.4
6.4 – 7.7
Blue Sky Influent
Blue Sky Effluent
7.1 – 7.5
6.0 – 7.3
Data Analysis and Calculations
11.1
12.
Process Ranges
Typical values for each plant are:
Town Branch Influent
Town Branch Effluent
None required.
Bibliography
12.1
U.S. EPA. Method 150.1 pH (Electrometric) Issued 1971 (Editorial revision 1978
and 1982). Methods for Chemical Analysis of Water and Wastes. EPA-600-4-79020; U.S. Environmental Protection Agency; Office of Research and
Development, Washington, DC, 1982.
12.2
HACH. USEPA Electrode Method 8156. DOC316.53.01245. Edition 5. HACH
Company, Loveland, CO, 2008.
12.3
Code of Federal Regulations (CFR). Guidelines Establishing Test Procedures for
the Analysis of Pollutants. 40 CFR 136.3, Title 40, Chapter 1. U.S. Environmental
Protection Agency; U.S. Environmental Protection Agency. pg 5-337. 2003.
11
SOP – WH Residual Chlorine
West Hickman WWTP Laboratory
Page 12 of 219
Revision Number 4
Last Revised 09/09
WH Total Residual Chlorine Analysis
HACH AutoCAT 9000 Total Chlorine Amperometric Forward Titration
procedure equivalent to
EPA Method 330.1 Chlorine, Total Residual (Titrimetric, Amperometric), Issued 1978
Standard Methods Part 4500-Cl D. (Chlorine Residual Amperometric Titration Method)
1.
Scope, Significance to Process and Application
1.1
2.
3.
Disinfection by chlorination is considered to be the primary mechanism for the
inactivation/destruction of pathogenic organisms in wastewater treatment plant
effluents and to prevent the spread of waterborne diseases to downstream users
and the environment. Final clarifier effluent is treated with chlorine as enters the
chlorine contact tanks allowing contact time for disinfection to transpire. Final
effluent is then treated with a dechlorinating agent reducing chlorine residual
concentration to within acceptable limits (see sec. 10.1 Permit limits). Complete
dechlorination is necessary to prevent chlorine related adverse effects on the
receiving environment. Town Branch Waste Water Treatment Plant uses Chlorine
Dioxide (ClO2) for chlorination and Sulfur Dioxide (SO2) as the dechlorinating
agent. Residual Chlorine analysis of treated plant effluent validates efficacy of
dechlorinating agent dosing and permit compliance.
Summary of Method
2.1
Executive Summary
West Hickman Laboratory uses a HACH AutoCAT 9000 autotitrator to perform
Residual Chlorine determinations. The AutoCAT 9000 bench top system
automatically completes all USEPA- approved amperometric titration methods for
chlorine, calculates analyte concentration, and provides real-time graphics
display. The AutoCAT’s forward amperometric titration procedure has a range of
0.0012 mg/L to 5.0 mg/L with an estimated detection limit of 0.0012 mg/L
2.2
Discussion
Chlorine (hypochlorite ion, hypochlorous acid) and chloramines liberate iodine
from potassium iodide at pH 4 or less in stoichiometeric proportions. The iodine
is titrated with a reducing agent phenylarsine and an amperometer detects the
endpoint. Although the actual measurement is that of the samples oxidation
potential, it is calculated and expressed as mg/L Cl because chlorine is the
dominating oxidizing agent present.
Health & Safety Precautions
3.1
3.2
Glassware involved, possible cut hazard.
Wastewater samples have the potential to be hazardous, use appropriate caution.
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West Hickman WWTP Laboratory
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4.
Sample Handling and Preservation
4.1
4.2
4.3
5.
Reagents
5.1
5.2
5.3
5.4
6.
Reagent 1 - Potassium Iodide 5%
Reagent 2 - pH 4.00 Buffer (Certified Acetic Acid). Both reagents are located
adjacent to the AutoCAT unit
Phenylarsine Oxide Standard Solution 0.00564 N
Chlorine Standard Solution, 25-30 mg/L as Cl2
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
7.
Residual chlorine is subject to dissipation by exposure to sunlight, mechanical
agitation, exchange of gases with the atmosphere and reaction with compounds in
the wastewater over time. For these reasons chlorine residuals should be analyzed
immediately (within 15 minutes of sampling time).
The sample should be taken gently into a glass 300 mL BOD bottle, completely
filling to above the base of the neck and installing the tapered glass stopper in a
manner that precludes air bubbles in the sample.
All glassware used in this method must have no chlorine demand, therefore do not
use plastic containers and pre-treat glassware accordingly. To remove chlorine
demand from clean glassware, soak in a dilute bleach solution (1 mL commercial
bleach to 1 liter of Nanopure water) for at least one hour. After soaking, rinse
thoroughly with Nanopure water. After analysis, thoroughly rinse all glassware
with Nanopure water to reduce the need for pretreatment.
HACH AutoCAT 9000 - Chlorine Amperometric Titrator
Beakers 250 mL
Graduated Cylinders 250 mL
1 mL fixed volume Finnpipette and 1 mL tips
Stirring bars.
Interferences
7.1
7.2
7.3
7.4
7.5
Accurate determinations of free chlorine cannot be made in the presence of
Nitrogen trichloride or Chlorine dioxide.
Some organic chloramines can also interfere.
Free halogens other than chlorine also will titrate as free chlorine.
Interference from copper has been noted in samples taken from copper pipe or
after heavy copper sulfate treatment of reservoirs.
Contamination of probe by metal ions such as copper, silver, iron interfere with
amperometric titrations. Fouled electrodes will not produce sharp endpoints.
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West Hickman WWTP Laboratory
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7.6
7.7
8.
Extended sample hold times, volatilization from mechanical agitation, and
exposure to various light sources can affect results.
At very low temperatures, there is slow response of cell and longer time is
required, but precision is not compromised.
Procedures
8.1
Steps
1) Prior to testing, pre-rinse all glassware and stir bars with sample (Do not
rinse with Nanopure once pre-rinsed).
2) Using a 250 mL graduated cylinder measure 200 mL of sample.
3) Pour sample into a 250 mL beaker with stirring bar, raise the electrode
assembly and place the beaker on unit.
4) Turn on instrument. The display will request user to press “1” for “Total Cl2
Fwd”, press “1”, display will then request user confirmation, press “1” to
confirm.
5) The display will request confirmation of sample volume (200 mL) press “1”
6) The sample will begin to stir.
7) Display will request the addition of 1 mL of Potassium Iodide 5% (Reagent 1),
pipette reagent into sample, then press “OK”
8) Display will request the addition of 1 mL of Acetate buffer pH 4 (Reagent 2),
pipette reagent into sample, then press “OK”
9) A mixing timer will countdown for 5 sec. then the display will request that the
electrodes be dipped into the sample, lower electrode assembly into sample and
press “OK”.
10) The display will request user to confirm the “Increment Setting” (should be
0.0010), press “1” to confirm.
11) Unit will begin analysis; total time required for analysis will vary with
sample strength and chosen increment value. During analysis the display
graphs the progress of the titration. Upon completion the unit will display
the resulting concentration value and calculated confidence limits, press
“OK” to accept results.
12) Record the results on the Total Chlorine Residual bench sheet. In the case of
the PTE sample, also log (in the provided location) the time sampled, time
received, and time analysis began. Note: If sample hold time (time sampled
to time analysis begins) exceeds 15 minutes the analysis is void and must be
rerun, beginning with resampling.
13) Select “END” if done with analysis or “Continue” to proceed to the next
sample to be analyzed.
Note: More detailed general information on the AutoCAT unit can be found in the
operator’s manual with details on the Forward Amperometric procedure starting on
page 101. The manual is located on the shelf adjacent to the AutoCAT unit.
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West Hickman WWTP Laboratory
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9.
8.2
Chlorine Standard Analysis
1) Obtain a Chlorine Standard Solution (25-30 mg/L) ampoule from “Chemical
Storage” fridge.
2) Carefully break top off ampoule.
3) Pipette 1.0 mL of standard into 1000 mL volumetric flask.
4) Bring to 1000 mL with Nanopure water.
5) Measure 200 mL of standard solution into beaker with stir bar.
6) Run titration as indicated in Section 8.1.
7) Record results in bench sheet.
8) The Laboratory Supervisor will determine if the standard is within the
expected range.
9) Measure 200 mL of Nanopure water into a clean 250 mL beaker and analyze
as indicated in Section 8.1 (This will be a Blank to confirm no chlorine
carry-over). Record results in bench sheet.
10) If chlorine is detected, re-run Blank until Below Detection Limit (BDL) is
obtained.
8.3
Helpful Hints
1) Analysis results are directly proportional to sample volumes therefore it is
very important that sample volume measurement is accurate.
2) Clean conditioned electrodes are required for the production of sharp, well
defined endpoints that are needed for precise analysis. Rinse electrodes
thoroughly before and after each use with Nanopure water, and store in
Nanopure water.
3) Routine use of the “Electrode Cleaning and Conditioning” procedure as
described in Section 9.1.4 of the Operator’s Manual will prevent problems.
4) Glassware must be clean and free of chlorine demand see section 4.3
QA/QC Requirements
9.1
A diluted standard (25-30 mg/L) and Blank(s) must be run once a week (See
Section 8.2).
9.2
5% of all samples must be run in duplicate.
9.3
Data acceptance criteria:
1) Results for the Standard must agree within 10% of the standard’s known
value.
2) Duplicate values must agree within 5%.
3) If these criteria are not met, corrective action is indicated. See Quality
Assurance Program (QAP) Sec. 15 “Corrective Action Policies and
Procedures”.
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West Hickman WWTP Laboratory
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10.
11.
Expected Results
10.1
KPDES Permit Requirements
KPDES Permit Limits on plant effluent residual chlorine at West Hickman
WWTP is a maximum monthly average of 0.011 mg/L, with a daily maximum
limitation of 0.019 mg/L. In the event that analysis results indicate values greater
then KPDES permit requirements, retest. If the value indicated by the retest is
greater than KPDES permit requirements, Immediately notify the Plant
Superintendent and Laboratory Supervisor.
10.2
Process Ranges
Expected residual chlorine results on plant effluent samples will be less than
0.010 mg/L, typically the results are BDL (below detection limit).
Data Analysis and Calculations
11.1
11.2
12.
Concentration values are read directly from the AutoCAT unit’s display, all
calculations are preformed internally.
The Laboratory Supervisor will determine if the results for the standard are within
the expected range.
Bibliography
12.1
U.S. EPA Method 330.3 Chlorine, Total Residual (Titrimetric, Amperometric)
Issued 1978. Methods for Chemical Analysis of Water and Wastes. EPA-600-479-020. U.S. Environmental Protection Agency; Office of Research and
Development, Washington, DC, 1982.
12.2
Standard Methods 4500-Cl D. Chlorine Residual Amperometric Titration Method.
APHA-American Public Health Association Standard Methods for the
Examination of Water and Wastewater; 21th edition ed.; American Water Works
Association and Water Pollution Control Federation: Washington, DC, 2005.
12.3
HACH AutoCat 9000 Chlorine Amperometric Titrator Instruction Manual.
HACH Company, Loveland, CO.
16
SOP – WH Fecal Coliforms
West Hickman WWTP Laboratory
Page 17 of 219
Revision Number 4
Last Revised 09/09
WH Fecal Coliform - Membrane Filter Procedure
U.S. EPA 600/8-78-017 Microbiological Methods for Monitoring the Environment:
Water and Wastes
1.
2.
3.
Scope, Significance to Process and Application
1.1
The fecal coliform analysis is applicable to investigations of stream pollution, raw
water sources, and wastewater treatment systems.
1.2
The fecal coliform analysis differentiates between coliforms of fecal origin.
Summary of Method
2.1
Executive Summary
The sample is filtered through a Millipore® membrane filter. The filter is placed
on a filter pad containing media in a sterile Petri dish. The samples are then
incubated at 44.5°C ± 0.2°C for 24 hours ± 2 hours. Colonies are counted and
fecal coliform calculations are performed.
2.2
Discussion
Fecal coliforms are defined as gram-negative, non-spore forming rods. The major
species is Escherichia coli, which indicates fecal pollution and the presence of
enteric pathogens. Colonies produced by fecal coliform bacteria are various
shades of blue. Non-fecal coliform colonies are gray to cream colored.
Health & Safety Precautions
3.1
3.2
3.3
3.4
4.
All municipal and industrial wastewaters are potentially hazardous. Gloves and
safety glasses should be worn when dispensing these samples.
Possible exposure to enteric pathogens. Care must be taken to avoid undue
exposure.
A flame is used to sterilize forceps. Maintain the area around the flame clear.
Contaminated (used) Petri dishes and lab equipment must be placed in
Biohazardous waste container. This Biohazardous waste container is autoclaved
before disposal.
Sample Handling and Preservation
4.1
4.2
4.3
Samples should be collected in clean, sterile glass or plastic containers.
If chlorine is in the sample, containers should be treated with 4 drops of
10% Sodium thiosulfate before autoclaving.
Run test immediately after sampling, or preserve sample at 4°C for a maximum of
6 hours.
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West Hickman WWTP Laboratory
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5.
Reagents
5.1
5.2
5.3
5.4
5.5
6.
Equipment & Lab Ware
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9
6.10
6.11
6.12
6.13
6.14
6.15
6.16
6.17
6.18
7.
Peptone powder
Peptone buffer solution pH 7.00 ± 0.1 (stored at 4°C)
m-FC media with rosolic acid for fecal coliforms (stored at 4°C)
Sodium thiosulfate 10% solution
Lysol disinfectant, 20% solution
Vacuum flask
Millipore® single use 47 mm Petri dishes with pads
Millipore® sterilized 47 mm filter
Forceps
4.5 X 9 inch sterile sampling bags
Bunsen burner and striker
Pipettes and sterile tips
Sterilized filter holder (plastic or glass)
Gable topped water bath at 44.5°C ± 0.2°C
Thermometer
ASTM Thermometer
Tower Steam Indicator Strips
ODO-Clave® Heat Activated Deodorant Pads
Autoclavable Biohazard waste bags and deposit box
Autoclave
Sterile blue sheets
Indicator tape
Autoclavable Nalgene® squeeze bottles for peptone
Interferences
7.1
7.2
7.3
Bacteria from the surrounding environment.
Cross contamination from one sample to the next.
Lack of aseptic techniques.
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West Hickman WWTP Laboratory
Page 19 of 219
Revision Number 4
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8.
Procedures
8.1
Steps
1) Clean work area with Lysol disinfectant, 20% solution.
2)
Light Bunsen burner with striker.
3)
Open sterile filter holder. Use sterile blue sheet as a sterile field. Indicator
trip and tape should indicate that the filter has been sterilized.
4)
Flame forceps and use it to remove the Indicator Strip without touching
anything else except the strip.
5)
Use Petri dishes with sterile pad already in dish.
6)
Break open ampule of media and pour onto media pad.
7)
Decant excess media and cover dish to protect sterile pad.
8)
Place the bottom of the sterile filter holder onto the vacuum flask.
9)
Flame forceps, remove sterilized filter from packaging and place onto
sterilized filter holder (grid side up). Do not touch the filter with anything
except the forceps.
10) Place or clamp the top unit onto filter holder.
11) Gently mix sample.
12) In advance, determine sample volume that will yield 20-60 fecal coliform
units (FCU).
13) If the volume of sample to be used is 0.1 to 5 mL, pour approximately 10
mL of peptone into filter unit before dispensing sample (Turn on vacuum
after the sample is introduced).
14) For sample volumes 5 to 50 mL, use sterile pipettes for dispensing into filter
unit.
15) Do not touch the inside of the filter holder unit. Do not allow the pipette tip
to touch the filter.
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West Hickman WWTP Laboratory
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16) For sample volumes 50 to 100 mL, pour sample into cylinder and use the
scale on the side of the cylinder for measurement.
17) Turn on vacuum.
18) Once sample has filtered through, turn off vacuum.
19) Rinse top of the filter unit with peptone two times using autoclaved peptone
in a Nalgene® squeeze bottle.
20) Turn on vacuum to drain peptone.
21) Turn off vacuum.
22) Flame forceps.
23) Remove top of the filter unit and place on the sterile blue field.
24) Open Petri dish.
25) Use sterile (flamed) forceps to grab the edge of the filter and remove it from
the filter holder unit.
26) Place filter, grid side up, onto edge of the Petri dish and gently slide it onto
the surface of the media saturated pad.
27) Replace Petri dish cover.
28) Place Petri dishes into a 4.5 X 9 inch sterile sampling bag. Make sure not
contaminate the inside of the bag.
29) Seal bag.
30) Place bag, with Petri dishes face down, into water bath at 44.5°C ± 0.2°C for
24 ± 2 hours.
31) Log initials, time, and date in the Microbiology bench sheet.
32) After 24 ± 2 hours, count blue colonies (See Section 10).
33) Log results, initials, time, and date in the Microbiology bench sheet.
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West Hickman WWTP Laboratory
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8.2
Before and After Blanks
1)
Blanks must be analyzed before and after filtration of a set of samples.
2)
Before any sample is filtered, place a sterile filter in the filter holder unit.
3)
Add 50 mL peptone buffer to filter unit.
4)
Turn on vacuum and filter the buffer, this will be the Before Blank.
5)
Remove and plate filter as indicated in Section 8.1.
6)
Proceed with sample filtration and plating.
7)
Run a Known Positive after all samples have been filtered (See Section 8.3).
8)
Place a sterile filter in the filter holder unit.
9)
Add 50 mL peptone buffer to filter unit.
10) Turn on vacuum and filter the buffer, this will be the After Blank.
11) Remove and plate filter as indicated in Section 8.1.
12) Log results, initials, time, and date in the Microbiology bench sheet.
8.3
Known Positive
1)
After all samples have been filtered, a Known Positive is filtered and plated
to ensure growth.
2)
Place a sterile filter in the filter holder unit.
3)
Add 10-20 mL peptone buffer to filter unit, then pipet 1.0 mL of mixed
liquor (or suitable sample with known fecal coliforms) into filter unit.
4)
Turn on vacuum and filter the sample, this will be the Known Positive.
5)
Remove and plate filter as indicated in Section 8.1.
6)
Log results, initials, time, and date in the Microbiology bench sheet.
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West Hickman WWTP Laboratory
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8.4
8.5
9.
Peptone Buffer Preparation
1)
Measure 1.0 g Peptone powder into 1L volumetric flask.
2)
Bring to volume with Nanopure water. Mix well.
3)
Pour Peptone buffer into Nalgene® autoclavable squeeze bottle.
4)
Take a sample of the buffer and measure pH, it should be 7.00 ± 0.1.
5)
Loosely screw caps/dispensers onto squeeze bottles.
6)
Autoclave Peptone buffer solutions.
7)
Allow to cool, then transfer squeeze bottles to Micro fridge.
Helpful Hints
1)
If the bacterial density of the sample is unknown, filter and plate out several
volumes or dilutions in order to achieve a countable density. The volumes
and/or dilutions should be expected to yield a countable membrane. In
addition, select two additional quantities representing one-tenth and ten
times this volume, respectively.
2)
Separate filter holder units may be required during a set of samples. These
will be indicated in the bench sheet.
3)
Do not use damaged or bent membrane filters.
4)
Rinse the filter unit thoroughly with Peptone buffer to avoid cross
contamination.
QA/QC Requirements
9.1
Before and After Blanks must be run with each set of samples tested.
9.2
One duplicate per test series must be run.
9.3
One “Known positive” must be run per test series.
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West Hickman WWTP Laboratory
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10.
Expected Results
10.1
KPDES Permit Requirements
200 CFU/100 mL for Monthly geometric mean (GED)
400 CFU/100 mL for Maximum Weekly GED
In the event that the GED is exceeds the KPDES permit requirements, notify the
Plant Superintendent and the Laboratory Supervisor.
10.2
11.
Data Analysis and Calculations
11.1
12.
Process Ranges
< 1 to >120,000 CFU/100 mL
See SOP – TB Fecal Coliforms for calculations.
Bibliography
12.1
U.S. EPA. Microbiological Methods for Monitoring the Environment: Water and
Wastes. EPA 600/8-78-017. U.S. Environmental Protection Agency;
Environmental Monitoring and Support Laboratory, Office of Research and
Development, Washington, DC, 1978. Page 124.
12.2
Kentucky Department for Environmental Protection, Kentucky Division of Water
and the Kentucky Division of Compliance Assistance. Discharge Monitoring
Report Manual. 2009. August 10, 2009 revision. 28 pp.
23
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West Hickman WWTP Laboratory
Page 24 of 219
Revision Number 5
Last Revised 09/09
WH Total Phosphorus Analysis
HACH Method 8190, PhosVer 3 with Acid Persulfate Digestion per EPA Method 365.2
1.
Scope, Significance to Process and Application
Phosphorus in wastewater is almost always present in the form of phosphates. There are
three major classes of phosphates; (1) Orthophosphates (fertilizer is major source), (2)
Polyphosphates (detergents and cleaning agents are major sources), and (3) Organic
Phosphates (biological waste is major source). Organic Phosphates are also formed from
orthophosphates during biological treatment of waste streams. Analysis of Total
Phosphorus includes all of the aforementioned forms of Phosphorus. The reduction of
Total Phosphorus levels throughout the wastewater treatment process is highly important,
as phosphorus concentrations in plant effluents must be low enough (See Sec.10.1,
Permit Requirements) to avoid detrimental effects on the receiving environment, such as
algae blooms.
2.
3.
Summary of Method
2.1
Executive Summary
Total phosphorus analysis at West Hickman Laboratory refers to the
spectrophotometric analysis of all phosphorus forms in a water/wastewater
sample that has been digested.
2.2
Discussion
West Hickman Lab uses, EPA approved, HACH Method 8190 with a HACH
DR/4000 spectrophotometer. Method range is 0.0 to 1.1 mg/L as Total P and 0.0
to 3.5 mg/L as PO43-, with an Estimated Detection Limit (EDL) of 0.06 mg/L
PO43-. The sample is first subjected to an acid persulfate digestion at 150˚C,
reducing all phosphorus forms to reactive form, which reacts with molybdate
reagent in an acid medium producing phosphomolybdate complex. This complex
is reduced with ascorbic acid forming a molybdenum blue color with intensity
proportional to the total phosphorus concentration, which is then quantifiable by
the spectrophotometer.
Health & Safety Precautions
3.1
3.2
3.3
During sample digestion, the digester block is HOT (150˚C), Burn Hazard. Use
digester block safety shields. In addition, the capped glass sample vials contain
hot (150˚C) acidic solution under pressure.
PhosVer3 powder pillow can be a respiratory hazard. Wear a mask or place
samples in hood when dispensing.
Wastewater samples should be considered potentially hazardous. Use proper
personal protective equipment.
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West Hickman WWTP Laboratory
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Last Revised 09/09
4.
Sample Handling and Preservation
4.1
5.
Reagents
5.1
5.2
5.3
5.4
5.5
5.6
6.
6.2
6.3
6.4
6.5
HACH DBR200 COD Reactor with Test’N Tube block calibrated to 150˚C, with
safety shields.
HACH DR/4000 Spectrophotometer with Test’N Tube adapter.
5 mL micropipetter with tips.
2 mL micropipetter with tips.
Microfunnel.
Interferences
7.1
7.2
8.
PhosVer 3 Phosphorus Reagent Powder.
Potassium Persulfate Reagent Powder.
Sodium Hydroxide Solution 1.56 N.
Total and Acid Hydrolyzable Test Vials (Test’N Tubes).
Nanopure Grade Water.
Phosphate Standard Solution 1 mg/L as PO43-.
Equipment & Lab Ware
6.1
7.
Analyze samples immediately after collection for best results. If prompt analysis
is impossible, preserve samples for up to 28 days by adjusting the pH to 2 or less
with H2SO4 and storing at 4˚C. Prior to analysis, allow samples to warm to room
temperature and neutralize. Document sample preservation.
Do not use commercial detergents containing phosphate for cleaning any lab ware
utilized in this method.
Excess Turbidity.
Procedures
8.1
Steps
1) Turn the HACH DRB200 COD reactor ON and preheat to 150˚ C by
pushing “Start”.
2)
Prepare a Reagent Blank by adding 5 mL of Nanopure lab water to a Total
and Acid Hydrolyzable Test Vial and label it “Reagent Blank”.
3)
Prepare a Low Range Standard by adding 1 mL of phosphate standard
solution (1 mg/L as PO43-) and 4 mL of Nanopure lab water to a Total and
25
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West Hickman WWTP Laboratory
Page 26 of 219
Revision Number 5
Last Revised 09/09
Acid Hydrolyzable Test Vial and label it “Low Standard” to produce a
standard of 0.2 mg/L PO43-.
4)
Prepare a High Range Standard by adding 5 mL of phosphate standard
solution (1 mg/L as PO43-) to a Total and Acid Hydrolyzable Test Vial and
label it “High Standard” to produce a standard of 1.0 mg/L PO43-.
5)
Put 5 mL of each sample to be analyzed in appropriately labeled Total and
Acid Hydrolyzable Test Vials. Make sure to save samples until samples are
read in case samples need to be diluted and/or reanalyzed.
6)
Using a microfunnel, add to each test vial the contents of one potassium
persulfate powder pillow, screw cap on tight, shake for 15 seconds, and
place into the COD reactor. Set the reactor timer to run for 30 minutes.
7)
CAREFULLY (vials are 150˚C) transfer hot vials to test tube racks and
allow to cool to ambient temperature before proceeding. Note: Vials are
under pressure until they cool (See Sec. 3.1).
8)
To each vial add 2 mL of sodium hydroxide solution 1.56N, cap and shake
to mix.
9)
Turn the DR/4000 ON and allow the unit to go through its startup and selfcheck routine. Press the soft key under “HACH PROGRAM”, type in the
number 3036 and press “ENTER”. The unit will respond by displaying
“HACH PROGRAM 3036 P Total TNT” and request to be zeroed.
10) To the blank, standards, and samples add to each the contents of one
PhosVer3 powder pillow, cap and shake for 15 seconds to mix, then allow a
2 minute reaction period before proceeding to step 11. Respiratory hazard
(See Sec. 3.2). (Note: Step 11 must be completed within 6 minutes of the
end of the 2 minute reaction period).
11) Place the reagent blank into the DR/4000 cell holder, close the lid and press
“ZERO”. Make sure the instrument is reading concentration and “FORM:
P”. One at a time, place all the vials (standards and samples) into the unit.
Note and record the indicated values on the bench sheet. If desired, during
the reading process, the soft arrow keys can be use to select units of P, PO43, or P2O5 although generally the “P” value (Total Phosphorus) will be
recorded.
26
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West Hickman WWTP Laboratory
Page 27 of 219
Revision Number 5
Last Revised 09/09
8.2
9.
Helpful Hints
1) Analysis results are directly proportional to sample volumes; therefore it is
very important that accurate sample volume measurements are made.
2)
The Total and Acid Hydrolyzable Test Vials are cuvettes to be analyzed
spectrophotometrically, and must provide a clear optical path. Prior to
reading, clean the vials by wiping them down a Kim Wipe moistened with
lab water, and then wipe them with a dry Kim Wipe.
3)
When washing lab ware involved with this method, use only phosphate free
cleaning agents, 1:1 HCL solution is recommended, followed by thorough
Nanopure lab water rinse. Note: Most of the lab ware used in this method is
disposable.
QA/QC Requirements
9.1
A Reagent Blank, High Standard (1.0 mg/L), and a Low Standard (0.2 mg/L)
must be run with every analytical run.
9.2
A total of 5% of all samples must be run in duplicate.
9.3
Data acceptance criteria:
9.3.1 Analysis values for Standards must agree within 10% of the Standards
known value or 0.12 mg/L, whichever is largest.
9.3.2 Duplicate values must agree within 5%.
If these criteria are not met, corrective action is indicated. See Quality Assurance
Program (QAP) Sec. 15 “Corrective Action Policies and Procedures”.
10.
Expected Results
10.1
KPDES Permit Requirements
1)
West Hickman Wastewater Treatment Plant effluent limitations are:
a. November 1st through April 30th - a monthly average of 1 mg/L and a
weekly average of 2 mg/L.
b. May 1st through October 31st - there are no effluent limitations on
Total Phosphorus.
Plant effluent composite samples are analyzed daily. Monthly and weekly
averages are reported.
In the event that analysis results indicate values greater than the KPDES permit
requirements, retest. If the value indicated by the retest is greater than KPDES
27
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West Hickman WWTP Laboratory
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Revision Number 5
Last Revised 09/09
permit requirements, Immediately notify the Plant Superintendent and the
Laboratory Supervisor.
10.2
11.
Data Analysis and Calculations
11.1
12.
Process Ranges
Typical phosphorus concentration values for plant influent vary primarily due to
rainfall. Within the treatment process phosphorus concentrations can vary due to
microbiological processes involving the release and uptake of phosphorus forms.
Typical values for each plant are:
West Hickman Influent 2 mg/L to 12 mg/L
West Hickman Effluent
<1.0 mg/L
Concentration values are read directly from the DR/4000 spectrophotometer.
Bibliography
12.1
HACH DR/4000 Procedure Manual, EPA approved Method 8190 Phosphorus,
Total. HACH Company, Loveland, CO.
12.2
Standard Methods Part 4500-P. Phosphorous. APHA-American Public Health
Association Standard Methods for the Examination of Water and Wastewater;
21th edition ed.; American Water Works Association and Water Pollution Control
Federation: Washington, DC, 2005.
12.3
U.S. EPA Method 365.2 Phosphorous, All Forms (Colorimetric, Ascorbic Acid,
Single Reagent) Revised March 1983. Methods for Chemical Analysis of Water
and Wastes. EPA-600-4-79-020. U.S. Environmental Protection Agency; Office
of Research and Development, Washington, DC, 1982.
28
APPENDIX D
LABORATORY BENCHSHEETS
D-1
TOWN BRANCH & WESTHICKMAN WASTEWATER TREATMENT PLANT
Town Branch Laboratory
Grab Samples
pH Method = EPA Method 150.1 pH (Electrometric) Issued 1971 (Editorial revision 1978 and 1982).
pH Meter Calibration
Temperature
First Buffer
Second Buffer
Third Buffer
=
°C
= 7.00 pH
= 4.00 pH
= 10.00 pH
Second Calibration
First Buffer
Second Buffer
Third Buffer
Calibrated By
Slope %
Calibrated By
Enter Date
= 7.00 pH
= 4.00 pH
= 10.00 pH
pH
1
2
3
4
5
6
7
8
9
10
11
12
13
Enter Date
14
15
Log Number
Sample Name
Date Samples Collected
Time Analysis Started
pH (Standard Units)
Analyst
Date Analysis Completed
KPDES Permit Limits on Plant Effluent
6.0 mg/L is the Lowest pH we can have on any given sample.
9.0 mg/L is the Highest pH we can have on any given sample.
Total Alkalinity, Orion Research Incorporated Laboratory Products Group
1
2
3
Log Number
Alkalinity
Sample Name
Blank
Standard
Date Samples Collected
Time Analysis Started
Volume of Sample (mL)
10.0
10.0
Dilution Factor = D
1.0
1.0
pH After Addition of Reagent
=A
Reading from Chart
Total Alkalinity = (A*D)
mg/L as CaCO3 - Blank
Analyst
Date Analysis Completed
PTE ph
Time sampled:_____________
Time received:_____________
Time started:_______________
Within 15 min limit: ________
Enter Date
4
5
Volatile Acids
6
7
Presevation: None required. Analyze Immediately.
Make Dilutions for all sample with Total Alkalinity > 225 mg/L CaCO3
Total Alkalinity Control Limits are from 93 to 107 mg/L on the 100mg/L Standard (Standards pH should be 4.41 ± 0.05 after reagent addition)
8
9
1
Log Number
Sample Name
Date Samples Collected
Time Analysis Started
V = Volume of Sample (mL)
N = Normality of the NaOH
A = Volume of NaOH Used
F = Recovery Factor
Volatile Acid
(A*N*60,000)/(V*F)
Analyst
Date Analysis Completed
Enter Date
2
3
D-2
TOWN BRANCH & WEST HICKMAN WASTEWATER TREATMENT PLANT
Town Branch Laboratory
Samples
Nitrogen, Ammonia = ULR TNT 830; HR TNT 832 /DR 5000
Nitrogen, Ammonia
Time
Analysis
Started
Log
Number
23213
23213
23146
23149
23153
23192
23194
23137
23138
23185
23186
23217
23218
23219
Sunday, October 18, 2009
Date:
Date
Sample
Collected
10/19/09
10/19/09
10/18/09
10/18/09
10/18/09
10/18/09
10/18/09
10/17/09
10/17/09
10/17/09
10/17/09
10/19/09
10/19/09
10/19/09
Sample
Reading
From
Spec.
Sample
Volume
Use
mg/L
mL
5.0
0.2
0.2
5.0
5.0
0.2
5.0
0.2
5.0
0.2
5.0
5.0
5.0
5.0
Low Standard 1.0 mg/L
High Standard 10.0 mg/L
TB Raw Influent Sun
TB Plant Effluent Sun
TB Creek Above Plant
WH Raw Influent Sun
WH Plant Effluent Sun
TB Raw Influent Sat
TB Plant Effluent Sat
WH Raw Influent Sat
WH Plant Effluent Sat
Above Leak
Below Leak
In Spring
Dilution
TNT
v/v
ULR
ULR
ULR
ULR
ULR
ULR
ULR
ULR
ULR
Analyst:
Quality Control Manager Is To Receive A Copy Of All "Q/C Results".
O
Presevation: Cool, 4 C H2SO4 to pH < 2. Maximum Holding Time 28 days.
EPA Limit
Averages
D-3
Monthly
EPA Limit
Weekly
TB PTE In Winter.
7 mg/L
10.5 mg/L
TB PTE In Summer.
2 mg/L
3 mg/L
WH PTE In Winter.
WH PTE In Summer.
Averages
Monthly
Weekly
10 mg/L
15 mg/L
4 mg/L
6 mg/L
TOWN BRANCH WASTEWATER TREATMENT PLANT
Town Branch Laboratory
BOD5 & CBOD5
Bottle Number
Volume of Seed Used in Seed Control
Initial D.O. of Seed Control Bottle
5-Day D.O. of Seed Control Bottle
=S
= B1
= B2
Carbonaceous Biochemical Oxygen Demand & Biochemical Oxygen Demand Method
EPA Method 405.1 (5-Days @ 20 ºC)
SEED CORRECTION DATA
BL1
BL2
BL3
Date Sample Collected:
1
2
3
( mL )
Date Sample Collected if Different from the above date:
( mg/L)
( mg/L)
Date Incubation
Started:
Sodium Sulfite Added
Date Incubation
Stopped:
Unseeded Blank:
Initial D.O.:
Winkler Method:
Initial D.O.:
5-Day D.O.:
Plant Effluent
BIOCHEMICAL OXYGEN DEMAND CALCULATIONS
Bottle Number
Log Number
Sample Name
Date Sample Collected
Time Analysis Started
Was Nitrification Inhibitor Used ( Y or N )
Sample Volume
=V
Initial D.O. of Sample
= D1
5-Day D.O. of Sample
= D2
( mL )
( mg/L )
( mg/L )
B.O.D. QC
Actual Value 198
mg/L
Analyst Setting Up Analysis:
D.O.
= Dissolved Oxygen
B.O.D = Biochemical Oxygen Demand
B.O.D.5 = 5-Day Biochemical Oxygen Demand
C.B.O.D.= Carbonaceous Biochemical Oxygen Demand
Analyst Reading & Calculating Analysis:
KPDES Permit Limits on Plant Effluent
10 mg/L for Monthly Average
QUALITY CONTROL MANAGER IS TO RECEIVE A COPY OF ALL QC RESULTS.
15 mg/L for Maximum Weekly Average
Presevation: Cool, 4OC. Maximum Holding Time 48 hours.
D-4
TOWN BRANCH WASTEWATER TREATMENT PLANT
Town Branch Laboratory
Grab Samples
Total Chlorine Residual
Analysis Method
Log Number
Sample Name
Date Samples Collected
Total Chlorine Residual
Analyst
Date Analysis Completed
Time Analysis Completed
1
2
3
4
5
6
7
8
EPA Method 330.1 Chlorine, Total Residual (Titrimetric, Amperometric) Issued 1978
Enter Date
9
10
11
12
13
14
15
16
EPA Method 330.5 Chlorine, Total Residual (Spectrophotometric, DPD) Issued 1978
(mg/L)
QUALITY CONTROL MANAGER IS TO RECEIVE A COPY OF ALL QC RESULTS.
Presevation: None required. Analyze Immediately.
KPDES Permit Limits on Plant Effluent
0.01 mg/L for Monthly Average
PTE Chlorine Residual
Time sampled:_____________ Within 15 min limit: ________
Time received:_____________
Time started:_______________
D-5
TOWN BRANCH WASTEWATER TREATMENT PLANT
Town Branch Laboratory
Grab Samples
Dissolved Oxygen Method = EPA Method 360.3 Oxygen, Dissolved (Luminescent Probe) Issued 2006.
Dissolved Oxygen Meter Calibration
Barometer Reading
Temperature, ºC:
Dissolved Oxygen Calibration Point
Dissolved Oxygen from HQ40D
Is Difference Less Than 0.2 Y or N
Calibrated By
Dissolved Oxygen
1
23172
Log Number
Sample Name
Date Samples Collected
Time Analysis Started
Dissolved Oxygen Reading
Analyst
Date Analysis Completed
2
23173
3
23176
R
PTE
A Creek
10/19/09
10/19/09
10/19/09
10/19/09
10/19/09
10/19/09
4
5
6
7
8
9
10
11
12
(mg/L)
QUALITY CONTROL MANAGER IS TO RECEIVE A COPY OF ALL QC RESULTS.
Presevation: None required. Analyze Immediately.
KPDES Permit Limits on Plant Effluent
7.0 mg/L is the Lowest Dissolved Oxygen we can have on any given sample.
PTE Dissolved Oxygen
Time sampled:_____________
Time received:_____________
Time started:_______________
Within 15 min limit: ________
13
14
15
D-6
TOWN BRANCH & WEST HICKMAN WASTEWATER TREATMENT PLANT
Town Branch Laboratory
Total Coliforms Method: EPA 600/8-78-017 Microbiological Methods for Monitoring the Environment: Water and Wastes, page 108
Fecal Coliforms Method: EPA 600/8-78-017 Microbiological Methods for Monitoring the Environment: Water and Wastes, page 124
Fecal Streptococci Method: EPA 600/8-78-017 Microbiological Methods for Monitoring the Environment: Water and Wastes, page 136
Microbiology
Log Number
Sample Name
Analysis
Sample Volume
Colonies Counted
Colonies /100 mL
=V
(mL)
=A
= (A/V) x 100
Averages
Date Sample Collected
Time Sample Collected
Time Sample Received by Laboratory
Date Analysis Started
Time Analysis Started
Set-up Analyst
Date Analysis Completed
Time Analysis Completed
Reading Analyst
1
Blank
Before
FC
50
2
23174
TB PTE
FC
10
3
23174
TB PTE
FC
10
4
5
7
8
9
23174
23177
Separate
23207
23207
23207
TB PTE TB A Crk Filter
WH PTE WH PTE WH PTE
FC
FC
FC
FC
FC
100
10
10
100
10
Known
Positive
FC
11
Blank
After
FC
50
01/00/00
01/00/00
01/00/00
01/00/00
01/00/00
01/00/00
01/00/00
01/00/00
01/00/00
01/00/00
01/00/00
01/00/00
01/00/00
01/00/00
01/00/00
01/00/00
01/00/00
01/00/00
01/01/00
01/00/00
01/01/00
01/01/00
01/01/00
01/01/00
01/01/00
01/01/00
01/01/00
01/01/00
01/01/00
01/01/00
12
13
Note: Plates should be read within 24hrs ± 2Hrs of set-up time.
QUALITY CONTROL MANAGER IS TO RECEIVE A COPY OF ALL QC RESULTS.
Total Coliforms Method
Fecal Coliforms Method
Fecal Strep. Method
Presevation: Cool, 4 C. Maximum Holding Time 6 hours.
KPDES Permit Limits on Plant Effluent
200/100mL for Monthly GED
400/100mL for Maximum Weekly GED
O
= TC
= FC
= FS
14
15
D-7
TOWN BRANCH & WEST HICKMAN WASTEWATER TREATMENT PLANT
Town Branch Laboratory
24 Hour Composite Samples
pH Method = EPA Method 150.1 pH (Electrometric) Issued 1971 (Editorial revision 1978 and 1982).
pH Meter Calibration
Temperature
First Buffer
Second Buffer
Third Buffer
=
°C
= 7.00 pH
= 4.00 pH
= 10.00 pH
Enter Date
= 7.00 pH
= 4.00 pH
= 10.00 pH
Second Calibration
First Buffer
Second Buffer
Third Buffer
Calibrated By
Slope %
Calibrated By
pH
1
2
3
4
5
6
7
8
9
10
11
12
Enter Date
13
14
Log Number
Sample Name
Date Samples Collected
Time Analysis Started
pH (Standard Units)
Analyst
Date Analysis Completed
KPDES Permit Limits on Plant Effluent
6.0 mg/L is the Lowest pH we can have on any given sample.
9.0 mg/L is the Highest pH we can have on any given sample.
Total Alkalinity, Orion Research Incorporated Laboratory Products Group
1
2
3
Log Number
Alkalinity
Sample Name
Blank
Standard
Date Samples Collected
Time Analysis Started
Volume of Sample (mL)
10.0
10.0
Dilution Factor = D
1.0
1.0
pH After Addition of Reagent
Reading from Chart
=A
Total Alkalinity = (A*D)
mg/L as CaCO3
Analyst
Date Analysis Completed
Presevation: None required. Analyze Immediately.
Total Hardness
4
1
Log Number
Sample Name
Date Samples Collected
Time Analysis Started
Volume of Sample = V (mL)
Molarity of the Titrant = M
mL of Titrant Used = A
Total Hardness = A
mg/L as Total Hardness as CaCO3
Analyst
Date Analysis Completed
2
3
D-8
TOTAL PHOSPHORUS ANALYSIS, HACH TNT +, DR 5000
Date
Sample
Collected
Time
Analysis
Started
No.
Log
Number
1
2
3
4
5
6
7
8
9
23216
23216
23146
23146
23148
23149
23171
23158
23154
Standard Low ( 0.75 mg/L )
Standard High ( 10.0 mg/L )
TB Raw Influent
TB Raw Influent Duplicate
TB Primary Effluent
TB Plant Effluent
Mixed Liquor
Return Activated Sludge
Raw Sludge Thickener Overflow
10
11
23192
23194
WH Raw Influent 1:10
WH PTE
12
13
23137
23138
TB Raw Influent Collected Sat
TB Plant Effluent Collected Sat
Analyst & Calibrated By:
Interferences:
Sample
Date:
PreDilution
Factor
Spec reading
Sample
Total
Volume in Phosphorus
Conc. in
mL
mg/L as P
LR
UHR
UHR
UHR
UHR
LR
LR
UHR
UHR
2.0
0.4
0.4
0.4
0.4
2.0
2.0
0.4
0.4
LR
LR
2.0
2.0
UHR
LR
0.4
2.0
Date of Analysis:
Saturday, January 00, 1900
Large amounts of turbidity may cause inconsistent results in the test because the
acid present in the powder pillows may dissolve some of the suspended particles
and because of variable desorption of orthophosphate from the particles.
Aluminum > 200 mg/L, Arsenate, Chromium > 100 mg/L, Copper > 10 mg/L, Iron > 100 mg/L, Nickel > 300 mg/L
extreme pH, Silica > 50 mg/L, Silicate > 10 mg/L, Sulfide > 90 mg/L, Zinc > 80 mg/L
D-9
TOWN BRANCH WASTEWATER TREATMENT PLANT
Town Branch Laboratory
24 Hour Composite Samples
Total Suspended Solids Method = Residue, Non-Filterable, EPA Method 160.2 (Gravimetric, Dried at 103 - 105 °C), Issued 1971.
Volatile Suspended Solids Method = Residue, Volatile, EPA Method 160.4 (Gravimetric, Ignition at 550 °C), Issued 1971.
Enter Date
Suspended Solids & Volatile Suspended Solids
1
2
3
4
5
6
7
8
9
10
11
Log Number
12
Sample Name
Additional Name Notations
Date Sample Collected
Time Analysis Started
Tare Number
Sample Volume
Wt. Tare & Dried Solids
Wt. Tare
Wt. of Solids
Total Suspended Solids
((W2 - W1)/V) x 1,000,000
Wt. Tare & Dried Solids
Wt. After 1 hr
Wt. Difference
=
=
=
=
V
W2
W1
W2 - W1
= TSS
(mL)
(gm)
(gm)
(gm)
(mg/L)
(gm)
= W2
(gm)
= W3
= W2 - W3 (gm)
Is Difference < 0.0005 grams?
Date Analyses Performed
Analyst Setting Up Analysis:
Analyst Reading & Calculating Analysis:
KPDES Permit Limits on Plant Effluent
30 mg/L for Monthly Average
45 mg/L for Maximum Weekly Average
Presevation: None required. Maximum Holding Time 7 days.
QUALITY CONTROL MANAGER IS TO RECEIVE A COPY OF ALL QC RESULTS.
13
14
D-10
TOWN BRANCH & WEST HICKMAN WASTEWATER TREATMENT PLANT
Town Branch Laboratory
24 Hour Composite Samples
Total Solids Method = Residue, Total, EPA Method 160.3 (Gravimetric, Dried at 103 - 105 °C), Issued 1971.
Total Volatile Solids Method = Residue, Volatile, EPA Method 160.4 (Gravimetric, Ignition at 550 °C), Issued 1971.
Total Solids & Volatile Solids
1
2
3
4
5
6
7
8
9
Enter Date
10
11
12
Log Number
Sample Name
Additional Name Notations
Date Sample Collected
Time Analysis Started
Tare Number
Wt. Tare & Wet Sample
Wt. Tare
Wt. of Wet Sample
Wt. Tare & Dried Solids
Wt. Tare
= W2
= W1
(gm)
(gm)
= W2 - W1 (gm)
= W3
= W1
(gm)
(gm)
= W3 - W1 (gm)
Wt. of Solids
Total Solids
= TS
(ppm)
((W3 - W1)/(W2 - W1)) x 1,000,000
% Total Solids
((W3 - W1)/(W2 - W1)) x 100
= TS
(%)
Wt. Tare & Dried Solids
Wt. Tare & Ashed Solids
= W3
= W4
(gm)
(gm)
= W3 - W4 (gm)
Wt. Volatilized Solids
Volatile Solids
= VS
(ppm)
((W3 - W4)/(W2 - W1)) x 1,000,000
Volatile Solids
((W3 - W4)/(W3 - W1)) x 100
= VS
(%)
Date Analyses Performed
QUALITY CONTROL MANAGER IS TO RECEIVE A COPY OF ALL QC RESULTS.
Analyst Setting Up Analysis:
Analyst Reading & Calculating Analysis:
13
D-11
TOWN BRANCH & WEST HICKMAN WASTEWATER TREATMENT PLANT
Town Branch Laboratory
Grab Samples
Total Suspended Solids Method = Residue, Non-Filterable, EPA Method 160.2 (Gravimetric, Dried at 103 - 105 °C), Issued 1971.
Volatile Suspended Solids Method = Residue, Volatile, EPA Method 160.4 (Gravimetric, Ignition at 550 °C), Issued 1971
Suspended Solids & Volatile Suspended Solids
1
2
3
4
5
6
7
8
9
10
11
Log Number
12
Sample Name
Additional Name Notations
Date Sample Collected
Time Analysis Started
Tare Number
Sample Volume
Wt. Tare & Dried Solids
Wt. Tare
Wt. Tare & Ashed Solids
Date Analyses Performed
=
=
=
=
V
W2
W1
W3
Analyst Setting Up Analysis:
(mL)
(gm)
(gm)
(gm)
QUALITY CONTROL MANAGER IS TO RECEIVE A COPY OF ALL QC RESULTS.
Presevation: None required. Maximum Holding Time 7 days.
Analyst Reading & Calculating Analysis:
Log-book/Monday/Tbmonsht.xls/TB Suspended Solids, Grab
Enter Date
13
14
15
D-12
TOWN BRANCH & WEST HICKMAN WASTEWATER TREATMENT PLANT
Town Branch Laboratory
24 Hour Composite Samples
Total Solids Method = Residue, Total, EPA Method 160.3 (Gravimetric, Dried at 103 - 105 °C), Issued 1971.
Total Volatile Solids Method = Residue, Volatile, EPA Method 160.4 (Gravimetric, Ignition at 550 °C), Issued 1971
Total Solids & Volatile Solids
1
2
3
4
5
6
7
8
9
Enter Date
10
11
Log Number
Sample Name
Additional Name Notations
Date Sample Collected
Time Analysis Started
Tare Number
Wt. Tare & Wet Sample
Wt. Tare
Wt. Tare & Dried Solids
Wt. Tare & Ashed Solids
Date Analyses Performed
= W2
= W1
= W3
= W4
(gm)
(gm)
(gm)
(gm)
QUALITY CONTROL MANAGER IS TO RECEIVE A COPY OF ALL QC RESULTS.
Analyst Setting Up Analysis:
Analyst Reading & Calculating Analysis:
Log-book/Monday/Tbmonsht.xls/TB Total Solids, 24Hr
12
13
D-13
TOWN BRANCH WASTEWATER TREATMENT PLANT
Town Branch Laboratory
Grab Samples
Total Solids Method = Residue, Total, EPA Method 160.3 (Gravimetric, Dried at 103 - 105 °C), Issued 1971.
Total Volatile Solids Method = Residue, Volatile, EPA Method 160.4 (Gravimetric, Ignition at 550 °C), Issued 1971
Total Solids & Volatile Solids
1
2
3
4
5
6
7
8
9
10
11
12
Enter Date
13
14
Log Number
Sample Name
Additional Name Notations
Date Sample Collected
Time Analysis Started
Tare Number
Wt. Tare & Wet Sample
Wt. Tare
Wt. Tare & Dried Solids
Wt. Tare & Ashed Solids
Date Analyses Performed
= W2
= W1
= W3
= W4
(gm)
(gm)
(gm)
(gm)
QUALITY CONTROL MANAGER IS TO RECEIVE A COPY OF ALL QC RESULTS.
Analyst Setting Up Analysis:
Analyst Reading & Calculating Analysis:
Log-book/Monday/Tbmonsht.xls/TB Total Solids, Grab
15
D-14
TOWN BRANCH WASTEWATER TREATMENT PLANT
Town Branch Laboratory
24 Hour Composite Samples
Settleable Matter Method = EPA Method 160.5 Settleable Matter (Volumetric, Imhoff Cone) Issued 1974
Settleable Matter
Enter Date
1
Log Number
Sample Name
Date Samples Collected
Vessel Used
2
3
Imhoff Cone Imhoff Cone Imhoff Cone
Time Analysis Started
Settleable Matter, 60 minutes = (mL/L/Hr.)
Analyst:
Date Analysis Completed:
QUALITY CONTROL MANAGER IS TO RECEIVE A COPY OF ALL QC RESULTS.
Presevation: None required. Maximum Holding Time 48 hours.
4
5
6
7
8
9
10
11
12
D-15
TOWN BRANCH WASTEWATER TREATMENT PLANT
Town Branch Laboratory
Grab Samples
Enter Date
Settleable Matter
1
Settleable Matter, 0 minutes = (mL/L)
5 minutes = (mL/L)
10 minutes = (mL/L)
15 minutes = (mL/L)
20 minutes = (mL/L)
25 minutes = (mL/L)
30 minutes = (mL/L)
Settlometer
800
Settled Sludge
Volume =
SSV(mL/L)
1000
40 minutes = (mL/L)
50 minutes = (mL/L)
60 minutes = (mL/L)
2 Hours = (mL/L)
3 Hours = (mL/L)
4 Hours = (mL/L)
0.0%
#DIV/0!
#DIV/0!
#DIV/0!
#DIV/0!
#DIV/0!
#DIV/0!
#DIV/0!
#DIV/0!
#DIV/0!
#DIV/0!
#DIV/0!
#DIV/0!
0.7
0.6
0.5
600
0.4
400
0.3
0.2
200
0.1
0
Average Mixed Liquor Total Suspended Solids
Average Mixed Liquor Volatile Suspended Solids
Sludge Volume Index (SVI) = (SSV30 x 1000)/MLSS
Sludge Density Index (SDI) = 100/SVI
Activated Sludge Zone Settling Rate Method = Method 2710 E. Zone
Settling Rate, WEF Standard Methods, 21st Edition.
0
0
5 10 15 20 25 30
40
50
60
Time (min)
Settled Sludge Volume Curve
Settled Sludge Concentration Curve
Power (Settled Sludge Concentration Curve)
Power (Settled Sludge Volume Curve)
Sludge Volume Index Method = Method 2710D. Sludge Volume Index,
WEF Standard Methods, 21st Edition.
Sludge Density Index Method = Method 2710D. Sludge Density Index,
WEF Standard Methods, 21st Edition.
Centrifuge Spin Test = Centrifuge Method for Estimating Suspended
Matter, WEF Simplified Laboratory Procedures for
Wastewater Examination, 3rd Edition, 1985, Pg 30.
Analyst
Date Analysis Completed
Rise Time
Activated Sludge Settled Volume Method = Method 2510 C. Settled
Volume, WEF Standard Methods, 21st Edition.
0.8
SSC (%)
1000
Settled Sludge
Concentration (SSC)
= SSCt = 1000 x
(ATC/SSVt)
1
0.9
Mixed Liquor
SSVt (mL/L)
Settled Sludge Time = SST(min)
Settled Sludge Curves
1200
Log Number
Sample Name
Date Samples Collected
Vessel Used
QUALITY CONTROL MANAGER IS TO RECEIVE A COPY OF
ALL QC RESULTS.
Centrifuge Spin Test
Sludge Volume after 15-minute Centrifuge Spin (mL)
Aeration Tank Mixed Liquor Concentration (ATC)
Analyst
Date Analysis Completed
Presevation: None required. Maximum Holding Time 48 hours.
D-16
Sunday’s Sheets
Town Branch Laboratory
pH Meter Calibration
Temperature
=
First Buffer
=
Second Buffer
=
Third Buffer
=
Slope
Calibrated By
Log Number
Sample Name
Date Samples Collected
Time Analysis Started
pH (Standard Units)
01/00/00
°C
7.00 pH
4.00 pH
10.00 pH
Dissolved Oxygen Meter Calibration
23127
PTE
01/00/00
23128
M/L
01/00/00
STD
01/00/00
Barometer Reading
Temperature, ºC:
Calculated Dissolved Oxygen Value
Calibrated By
Log Number
Sample Name
Date Samples Collected
04/26/63
0000
PTE
04/26/63
Time Analysis Started
Dissolved Oxygen Reading, mg/L
Analyst
Analyst
Date Analysis Completed
01/00/00
01/00/00
01/00/00
04/26/63
Date Analysis Completed
Presevation: None required. Analyze Immediately.
KPDES Permit Limits on Plant Effluent pH
6.0 mg/L is the Lowest pH we can have on any given sample.
9.0 mg/L is the Highest pH we can have on any given sample.
KPDES Permit Limits on Plant Effluent DO
7.0 mg/L is the Lowest Dissolved Oxygen
we can have on any given sample.
pH Method = EPA Method 150.1 pH (Electrometric) Issued
Dissolved Oxygen Method = EPA Method 360.1 Oxygen,
1971 (Editorial revision 1978 and 1982).
Dissolved (Membrane Electrode) Issued
1971
Fill in yellow squares.
Fill in yellow squares.
Charlie Begley 234 - 4886
Mark Stager 368 - 7296
Charlie Begley 234 - 4886
Mark Stager 368 - 7296
D-17
Sunday’s Sheets
Total Chlorine Residual
Microbiology
Analysis Method
Log Number
Sample Name
Analysis
Sample Volume
= V (mL)
Colonies Counted = A
Colonies /100 mL
Blank Before
50
14969
23129
PTE
PTE
Fecal Coliform
100
10
Blank After
50
Total Chlorine Residual
Saturday, January 00, 1900
Time Sample Collected
Time Analysis Started
(mg/L)
Analyst
Date Analysis Completed
Time Analysis Completed
Set-up Analyst
Date Analysis Completed
Time Analysis Completed
Reading Analyst
0000
PTE
04/26/63
Log Number
Sample Name
Date Samples Collected
Time Analysis Started
= (A/V x 100)
Averages:
Date Sample Collected
04/26/63
EPA Method 330.1 Chlorine,
Total Residual (Titrimetric,
Amperometric) Issued 1978
B.D.L. = Below Detection Limit = < 0.01 mg/L
Sunday, January 01, 1900
O
Preservation: Cool, 4 C. Maximum Holding Time 6 hours.
Note: Plates should be read within 24hrs ± 2Hrs of set-up time.
KPDES Permit Limits on Plant Effluent Fecal Coliform
200/100mL for Monthly GED
400/100mL for Maximum Weekly GED
KPDES Permit Limits on Plant Effluent Res. Cl2
0.010 mg/L for Monthly Average
0.019 mg/L for Daily Maximum
Presevation: None required. Analyze Immediately.
04/26/63
D-18
Waterproof TDSTestr and ECTestr Series Instructions
Before you Begin
Changing Batteries
Electrode replacement:
Remove electrode cap. Soak electrodes for a few
minutes in alcohol to remove oils.
1. Open battery compartment lid (end with
+
You can replace the electrode module at the fraction
of the cost of a new Testr. When the Testr fails to
calibrate, gives fluctuating readings in buffers,
shows error messages ‘E2’ or ‘OR’ in a buffer, and
the procedures in the Maintenance section do not
help, you need to change the electrode.
–
1. With dry hands, grip the ribbed Testr collar
2. Remove old batteries;
Calibration
Tester is factory calibrated. However, to ensure
accuracy, calibrate on a regular basis.
Select a calibration standard appropriate for
your Testr:
TDSTestr Low: from 200 to 1990 ppm
TDSTestr High: from 2.00 to 10.00 ppt
ECTestr Low: from 200 to 1990 µS
ECTestr High: from 1.00 to 19.90 mS
It is best to select a standard close to the test solution value.
1. Open battery compartment
INC
lid (end with lanyard loop).
The two white buttons are
Increment (INC) and
Decrement (DEC)
calibration keys.
2. Rinse electrode in
lanyard loop).
replace with fresh ones.
Note polarity (shown in
battery compartment
and in picture at right).
3. Recalibrate after battery
change.
keypad
Tester Maintenance
• To improve performance, clean the electrodes
by rinsing them in alcohol for 10-15 minutes.
Remove white plastic cup insert to clean viscous solutions.
• Replace all batteries if low battery indicator
appears, or if readings are faint or unstable.
• If you experience drift, periodically let electrode fully dry.
When you need a new electrode, see “Electrode
Replacement” at right.
DEC
deionized water, then
rinse it in calibration standard,
then dip it into a container of calibration
standard.
3. Switch unit on (ON/OFF key). Wait several
minutes for display to stabilize.
4. Press the INC or DEC keys to adjust reading to
match the calibration standard value.
5. After 3 seconds without a key press, the
display flashes 3 times, then shows “ENT”.
The tester accepts calibration value; returns to
measurement mode.
6. Replace battery cap.
TDS or Conductivity Testing
1. Remove electrode cap. Switch unit on
(ON/OFF key).
2. Dip electrode into test solution. Make sure
sensor is fully covered.
3. Wait for reading to stabilize (Automatic
Temperature Compensation corrects for
temperature changes). Note reading.
4. Press ON/OFF to turn off Tester. Replace
electrode cap. Note: Tester automatically shuts
off after 8.5 minutes of nonuse.
Specifications
Testr
Range
Resolution
Accuracy
TDS Factor
Calibration
Standard
Range
0 to
1990
ppm
0 to
10.00
ppt
10 ppm 0.10 ppt
0 to
1990
µS
0 to
19.90
mS
A
Rotate collar
away from you
Ribbed Collar
2. Pull the old electrode module away from the
Testr.
3. Align the four tabs on the new module so
they match the four slots on the testr. (see
diagram B).
B
Small O-Ring
(hidden)
Electrode
module
Large
O-Ring
10 µS 0.10 mS
±1% full scale
0.4 to 1.0
selectable
200 to
1990
ppm
2.00 to
10.00
ppt
—
200 to 1.00 to
1990 19.90
µS
mS
Calibration 1 point (calibration range is ±30%
of factory default parameter)
Insert Electrode
Small
Tab
Large Tab
4. Gently push the module onto the slots to seat
it in position. Push the smaller O-ring fully
onto the new electrode module. Push the collar over the module and thread it into place
by firmly twisting clockwise.
ATC
0 to 50°C (2% per °C)
Warranty:
Operating
Temp.
0 to 50ºC
Each TDSTestr and ECTestr meter body is warranted against defects in materials and workmanship for a period of 12 months from the date
of purchase; the electrode module is warrantied
for a period of 6 months from the date of purchase. If repair, adjustment or replacement is
necessary and has not been the result of abuse or
misuse within the 6 month period, please return
the Testr—freight pre-paid—and correction will
be made without charge. Out of warranty products will be repaired on a charge basis.
Power
HOLD function
Press HOLD key to freeze display. Press HOLD
again to release.
TDS Low TDS Hi EC Low EC Hi
with electrode facing you. Twist the collar
counter clockwise. (see diagram A). Save the
ribbed Testr collar and O-ring for later use.
Four 1.5V alkaline batteries
(Eveready A76BP; supplied)
150 hrs. continuous use
Alternate replacement Model
Eveready 303 silver oxide,
70 hrs. continuous use.
Dimensions 6.5"L x 1.5" dia. (165 x 38mm)
Weight
3.25 oz (90 gms)
Setting TDS Factor (TDSTestrs only)
Return of Items:
The TDSTestrs let you select a TDS factor of
0.4 to 1.0.
Authorization must be obtained from your
OAKTON Distributor before returning items for
any reason. When applying for authorization,
please include information regarding the reason
the item(s) are to be returned.
1. Open battery compartment. With meter on,
press the HOLD key, then press the INC key
(INC key is inside battery compartment; see
diagram at left).
2. Press the INC or DEC keys to adjust the TDS
factor.
3. After 3 seconds without a key press, the display flashes 3 times, then shows “ENT”. Tester
accepts TDS factor and returns to measurement mode.
4. Replace battery cap.
Note: We reserve the right to make improvements in design, construction and appearance of
products without notice. Prices are subject to
change without notice.
Dissolved
Oxygen
Water Quality Test Kit
Instruction Manual • Code 7414/5860
INTRODUCTION
Aquatic animals need dissolved oxygen to live. Fish, invertebrates, plants, and
aerobic bacteria all require oxygen for respiration. Oxygen dissolves readily
into water from the atmosphere until the water is saturated. Once dissolved in
the water, the oxygen diffuses very slowly and distribution depends on the
movement of the aerated water. Oxygen is also produced by aquatic plants,
algae, and phytoplankton as a by-product of photosynthesis.
The amount of oxygen required varies according to species and stage of life.
Dissolved Oxygen levels below 3 ppm are stressful to most aquatic organisms.
Dissolved Oxygen levels below 2 or 1 ppm will not support fish. Levels of 5 to
6 ppm are usually required for growth and activity.
This test kit uses the azide modification of the Winkler method for
determining dissolved oxygen.
TABLE OF CONTENTS
Page
Kit Contents....................................................................... 2
Test Procedure
Part 1: Collecting a Water Sample ..................................... 3
Part 2: Adding the Reagents ............................................... 4
Part 3: Titration................................................................... 5
EPA Compliance................................................................. 8
Dissolved Oxygen Fact Sheet............................................. 10
General Safety Precautions................................................ 13
Use Proper Analytical Techniques..................................... 14
Material Safety Data Sheets ............................................. 15
Kit Diagrams..................................................................... 21
Short Form Instructions ...................................... Back Cover
WARNING! This set contains chemicals
that may be harmful if misused. Read
cautions on individual containers
carefully. Not to be used by children
except under adult supervision
2
KIT CONTENTS
QUANTITY
30 mL
30 mL
50 g
30 mL
60 mL
30 mL
1
1
1
1
CONTENTS
CODE
*Manganous Sulfate Solution
*Alkaline Potassium Iodide Azide
*Sulfamic Acid Powder (7414 Kit)
*Sulfuric Acid, 1:1 (5860 Kit)
*Sodium Thiosulfate, 0.025N
Starch Indicator Solution
Spoon, 1.0 g, plastic (7414 Kit)
Direct Reading Titrator
Test Tube, 5-10-12.9-15-20-25 mL,
glass, w/cap
Water Sampling Bottle, 60 mL, glass
*4167-G
*7166-G
*6286-H
*6141WT-G
*4169-H
4170WT-G
0697
0377
0608
0688-DO
*WARNING: Reagents marked with a * are considered to be potential health
hazards. To view or print a Material Safety Data Sheet (MSDS) for these
reagents see MSDS CD or www.lamotte.com. To obtain a printed copy,
contact LaMotte by email, phone or fax.
To order individual reagents or test kit components, use the specified code
numbers.
3
TEST PROCEDURE
PART 1 - COLLECTING THE WATER SAMPLE
2.
1.
Rinse the Water Sampling Bottle
(0688-DO) with the sample water.
Tightly cap the bottle, and
submerge it to the desired depth.
3.
4.
Remove the cap and allow the
bottle to fill.
Tap the sides of the bottle to
dislodge any air bubbles.
5.
6.
Retrieve the bottle and make
sure that no air bubbles are
trapped inside.
Replace the cap while the
bottle is still submerged.
4
TEST PROCEDURE
PART 2 - ADDING THE REAGENTS
NOTE:
1.
2.
Remove the cap
from the bottle.
Be careful not to
introduce air into the
sample while adding
the reagents.
3.
Cap the bottle and mix
by inverting several
times. A precipitate will
form.
Immediately add
8 drops of
*Manganous
Sulfate Solution
(4167) AND
Add 8 drops of
*Alkaline
Potassium Iodide
Azide (7166).
4.
Allow the precipitate
to settle below the
shoulder of the bottle.
5.
For Kit Code 7414:
Immediately use the 1.0 g
spoon (0697) to add one
level measure of *Sulfamic
Acid Powder (6286).
OR
Cap and gently invert the bottle
to mix the contents until the
precipitate and the reagent
have totally dissolved. The
solution will be clear yellow
to orange if the sample
contains dissolved oxygen.
6.
5
For Kit Code 5860:
Add 8 drops of
*Sulfuric Acid, 1:1
(6141WT).
At this point
the sample has
been "fixed" and contact
between the sample and the
atmosphere will not affect the
test result. Samples may be
held at this point and titrated
later.
NOTE:
TEST PROCEDURE
PART 3 - THE TITRATION
1.
2.
Fill the titration
tube (0608) to the
20 mL line with the
fixed sample. Cap
the tube.
Depress plunger
of the Titrator
(0377).
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
3.
0
0.1
0.2
Insert the Titrator
into the plug in the
top of the *Sodium
Thiosulfate, 0.025N
(4169) titrating
solution.
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
the bottle and
4. Invert
slowly withdraw the
NOTE:
plunger until the
large ring on the
plunger is opposite
the zero (0) line on
the scale.
If small air bubbles appear in
the Titrator barrel, expel them
by partially filling the barrel
and pumping the titration
solution back into the reagent
container. Repeat until bubble
disappears.
1.0
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
the bottle
5. Turn
upright and remove
the Titrator.
NOTE:
0
0
0.1
0.1
0.2
0.2
0.3
0.3
0.4
0.4
0.5
0.5
0.6
0.6
0.7
0.7
0.8
0.8
0.9
0.9
1.0
1.0
If the sample is a very
pale yellow, go to Step 9.
continued . . .
6
TEST PROCEDURE
6.
Insert the tip of the
Titrator into the
opening of the
titration tube cap.
8.
7.
Slowly depress the
plunger to dispense the
titrating solution until
the yellow-brown
color changes to a very
pale yellow. Gently
swirl the tube during
the tiration to mix the
contents.
0
0.1
0.2
0
0.3
0.1
0.4
0.2
0.5
0.3
0.6
0.4
0.7
0.5
0.8
0.6
0.9
0.7
1.0
0.8
0.9
1.0
9.
0
0.1
0.2
Carefully remove the
Titrator and cap. Do
not to disturb the
Titrator plunger.
0.3
Add 8 drops of
Starch Indicator
Solution
(4170WT). The
sample should
turn blue.
0.4
0.5
0.6
0.7
0.8
0.9
1.0
10.
11.
0
0.1
0.2
Cap the titration
tube. Insert the tip of
the Titrator into the
opening of the
titration tube cap.
0.3
0.4
0.5
0.6
0.7
0
0.8
0.1
0.9
0.2
1.0
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
12.
Result:
4.0 ppm
Read the test result directly
from the scale where the large
ring on the Titrator meets the
Titrator barrel. Record as ppm
Dissolved Oxygen. Each
minor division on the Titrator
scale equals 0.2 ppm.
Continue
titrating until
the blue color
disappears and
the solution
becomes
colorless.
4.0
5.0
7
TEST PROCEDURE
NOTE:
If the plunger ring reaches the bottom line on the scale (10 ppm)
before the endpoint color change occurs, refill the Titrator and
continue the titration. Include the value of the original amount
of reagent dispensed (10 ppm) when recording the test result.
NOTE:
0
When testing is complete, discard titrating
solution in Titrator. Rinse Titrator and titration
tube thoroughly. DO NOT remove plunger or
adapter tip.
8
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
EPA COMPLIANCE
To qualify as an EPA accepted test, and to achieve the greatest accuracy, the
Sodium Thiosulfate Solution, 0.025N (4169) must be standardized daily. This
procedure follows Standard Methods for the Examination of Water and
Wastewater. Numbers in ( ) are for LaMotte products. These products are not
included in this kit but can be ordered from LaMotte Company by using the
specified code number.
1.
2.
0
0.1
Use a 10 mL graduated
cylinder (0416) to add
15 mL of Deionized
Water (5115) to the
titration tube (0608).
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
3.
4.
Add 2 drops
of Sulfuric
Acid, 5N
(8517WT).
Use the 0.1 g
spoon (0699) to
add 0.2 g
Potassium Iodide
Crystals (6809).
6.
Swirl to dissolve.
Solution will turn
yellowish brown.
1.0
0.9
0.8
0.5
0.4
0.3
0.2
0.1
0
9
0.6
Fill another Direct
Reading Titrator
(0376) with Sodium
Thiosulfate Solution,
0.025N (4169).
0.7
5.
Use a Direct
Reading Titrator,
0-1 Range (1.0
mL capacity)
(0376) to add 2
mL of Potassium
Bi-iodate (7346).
EPA COMPLIANCE
7.
While gently swirling the
tube, add Sodium
Thiosulfate, 0.025N until
the color fades to pale
yellow. It will be
necessary to refill the
Direct Reading Titrator.
8.
Add 3 drops of
Starch
Indicator
Solution
(4170WT).
The solution
will turn blue.
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
9.
Continue adding Sodium
Thiosulfate, 0.025N until
the blue color disappears
and the solution is
colorless.
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
10.
Result:
2.0 ppm
Read the test result directly from the scale
where the large ring on the Titrator meets the
Titrator barrel. Include the value of the original
amount dispensed (1 mL). If the reading is 2.0
+/-0.1 mL, the Sodium Thiosulfate, 0.025N
(4169) is satisfactory. If not, discard and
replace with new reagent.
10
2.0
3.0
DISSOLVED OXYGEN FACT SHEET
Oxygen is critical to the survival of aquatic plants and animals, and a shortage
of dissolved oxygen is not only a sign of pollution, it is harmful to fish. Some
aquatic species are more sensitive to oxygen depletion than others, but some
general guidelines to consider when analyzing test results are:
5–6 ppm
<3 ppm
<2 ppm
Sufficient for most species
Stressful to most aquatic species
Fatal to most species
Because of its importance to the fish’s survival, aquaculturists, or “fish farmers,”
and aquarists use the dissolved oxygen test as a primary indicator of their
system’s ability to support healthy fish.
WHERE DOES THE OXYGEN COME FROM?
The oxygen found in water comes from many sources, but the largest source is
oxygen absorbed from the atmosphere. Wave action and splashing allows more
oxygen to be absorbed into the water. A second major source of oxygen is
aquatic plants, including algae; during photosynthesis plants remove carbon
dioxide from the water and replace it with oxygen.
Absorption
Oxygen is continuously moving between the water and surrounding air. The
direction and speed of this movement is dependent upon the amount of
contact between the air and water. A tumbling mountain stream or
windswept, wave-covered lake, where more of the water’s surface is exposed to
the air, will absorb more oxygen from the atmosphere than a calm, smooth
body of water. This is the idea behind aerators: by creating bubbles and waves
the surface area is increased and more oxygen can enter the water.
Photosynthesis
In the leaves of plants, one of the most important chemical processes on Earth
is constantly occurring: photosynthesis. During daylight, plants constantly
take carbon dioxide from the air, and in the presence of water convert it to
oxygen and carbohydrates, which are used to produce additional plant
material. Since photosynthesis requires light, plants do not photosynthesize at
night, so no oxygen is produced. Chemically, the photosynthesis reaction can
be written as:
Light
+
nCO2 + nH2O
Light
+ Carbon + Water
Dioxide
¾¾®
(C2HO)n
+
nO2
¾
¾® Carbohydrate + Oxygen
11
WHERE DOES THE OXYGEN GO?
Once in the water, oxygen is used by the aquatic life. Fish and other aquatic
animals need oxygen to breathe or respire. Oxygen is also consumed by
bacteria to decay, or decompose, dead plants and animals.
Respiration
All animals, whether on land or underwater, need oxygen to respire, grow and
survive. Plants and animals respire throughout the night and day, consuming
oxygen and producing carbon dioxide, which is then used by plants during
photosynthesis.
Decomposition
All plant and animal waste eventually decomposes, whether it is from living
animals or dead plants and animals. In the decomposition process, bacteria use
oxygen to oxidize, or chemically alter, the material to break it down to its
component parts. Some aquatic systems may undergo extreme amounts of
oxidation, leaving no oxygen for the living organisms, which eventually leave
or suffocate.
OTHER FACTORS
The oxygen level of a water system is not only dependent on production and
consumption. Many other factors work together to determine the potential
oxygen level, including:
• Salt vs. fresh water - Fresh water can hold more oxygen than salt
water.
• Temperature - Cold water can hold more oxygen than warm water.
• Atmospheric pressure (Altitude) - The greater the atmospheric
pressure the more oxygen the water will hold.
TESTING DISSOLVED OXYGEN
Dissolved oxygen is often tested using the Azide modification of the Winkler
method. When testing dissolved oxygen it is critical not to introduce
additional oxygen into the sample. Many people avoid this problem by filling
the sample bottle all the way and allowing the water to overflow for one
minute before capping.
The first step in a DO titration is the addition of Manganous Sulfate Solution
(4167) and Alkaline Potassium Iodide Azide Solution (7166). These reagents
react to form a white precipitate, or floc, of manganous hydroxide, Mn(OH)2.
Chemically, this reaction can be written as:
MnSO4
+
2KOH
Manganous + Potassium
Sulfate
Hydroxide
¾
¾®
Mn(OH)2
¾¾®
Manganous + Potassium
Hydroxide
Sulfate
12
+
K2SO4
Immediately upon formation of the precipitate, the oxygen in the water
oxidizes an equivalent amount of the manganous hydroxide to brown-colored
manganic hydroxide. For every molecule of oxygen in the water, four
molecules of manganous hydroxide are converted to manganic hydroxide.
Chemically, this reaction can be written as:
4Mn(OH)2 +
O2
+
2H2O
¾¾®
4Mn(OH)3
Manganous +
Hydroxide
Oxygen
+
Water
¾¾®
Manganic
Hydroxide
After the brown precipitate is formed, a strong acid, such as Sulfamic Acid
Powder (6286) or Sulfuric Acid, 1:1 (6141) is added to the sample. The acid
converts the manganic hydroxide to manganic sulfate. At this point the
sample is considered “fixed” and concern for additional oxygen being
introduced into the sample is reduced. Chemically, this reaction can be
written as:
2Mn(OH)3 + 3H2SO4
¾¾®
Manganic
Hydroxide
¾
¾®
+ Sulfuric
Acid
Mn2(SO4)3 +
Manganic
Sulfate
+
6H2O
Water
Simultaneously, iodine from the potassium iodide in the Alkaline Potassium
Iodide Azide Solution is oxidized by manganic sulfate, releasing free iodine
into the water. Since the manganic sulfate for this reaction comes from the
reaction between the manganous hydroxide and oxygen, the amount of iodine
released is directly proportional to the amount of oxygen present in the
original sample. The release of free iodine is indicated by the sample turning a
yellow-brown color. Chemically, this reaction can be written as:
Mn2(SO4)3 +
2KI
¾
¾® 2MnSO4 +
K2SO4
+
I2
Manganic + Potassium ¾
¾® Manganous + Potassium + Iodine
Sulfate
Iodide
Sulfate
Sulfate
The final stage in the Winkler titration is the addition of sodium thiosulfate.
The sodium thiosulfate reacts with the free iodine to produce sodium iodide.
When all of the iodine has been converted the sample changes from
yellow-brown to colorless. Often a starch indicator is added to enhance the
final endpoint. Chemically, this reaction can be written as:
2Na2S2O3
+
I2
¾¾®
Na2S4O6
+
2NaI
Sodium
Thiosulfate
+
Iodine
¾¾®
Sodium
Tetrathionate
+
Sodium
Iodide
13
GENERAL SAFETY PRECAUTIONS
1.
2.
tion
Instruc l
u
n
Ma a
Store the test kit
in a cool, dry
area.
3.
Read the labels on
all reagent bottles.
Note warnings and
first aid information.
Read all Material
Safety Data Sheets.
4.
al
Materi
Safety
Data
Sheet
contact between
5. Avoid
reagent chemicals and skin,
6.
eyes, nose, and mouth.
7.
Read all
instructions and
note precautions
before performing
the test
procedure.
Keep all
equipment
and reagent
chemicals out
of the reach of
young
children.
Wear safety glasses when
performing test procedures.
In the event of an accident or suspected poisoning, immediately
call the Poison Center phone number in the front of your local
telephone directory or call a physician. Additional information
for all LaMotte reagents is available in the United States,
Canada, Puerto Rico, and the US Virgin Islands from Chem-Tel
by calling 1-800-255-3924. For other areas, call 813-248-0585
collect to contact Chem-Tel’s International access number. Each
reagent can be identified by the four digit number listed on the
upper left corner of the reagent label, in the contents list and in
the test procedures.
14
USE PROPER ANALYTICAL TECHNIQUES
1.
3.
5.
dropper bottles vertically
2. Hold
upside-down, and not at an
Use test tube caps or
stoppers, not your
fingers, to cover
tubes during
shaking or mixing.
angle, when dispensing a
reagent. Squeeze
the bottle gently to
dispense the
reagent one drop
at a time.
4.
Wipe up any reagent
chemical spills immediately.
6.
Tightly close all
containers immediately
after use. Do not
interchange caps from
containers.
15
Thoroughly rinse test tubes
before and after each test.
Avoid
prolonged
exposure of
equipment and
reagents to
direct sunlight.
Protect
reagents from
extremes of
temperature.
DISSOLVED OXYGEN KIT · CODE 7414
Instructions
4170
WT-G
4169-H
6286-H
0608
Tube
7166-G
0688-DO
4167-G
0377
0697
DISSOLVED OXYGEN KIT · CODE 5860
Instructions
4169-H
4170
WT-G
6141
WT-G
0608
Tube
7166-G
0688-DO
4167-G
0377
16
17
SHORT FORM INSTRUCTIONS
Read all instructions before performing test. Use this guide as
a quick reference.
1.
Fill Water Sampling Bottle (0688-DO).
2.
Add 8 drops of *Manganous Sulfate Solution (4167).
3.
Add 8 drops of *Alkaline Potassium Iodide Azide (7166).
4.
Cap and mix.
5.
Allow precipitate to settle.
6.
7.
Use the 1.0 g spoon to add *Sulfamic Acid Powder (6286) or
add 8 drops of Sulfuric Acid, 1:1 (6141WT).
Cap and mix until reagent and precipitate dissolve.
8.
Fill test tube (0608) to the 20 mL line.
9.
Fill Titrator with *Sodium Thiosulfate, 0.025N (4169).
10. Titrate until sample color is pale yellow. DO NOT DISTURB
TITRATOR.
11. Add 8 drops of Starch Indicator (4170WT).
12. Continue titration until blue color just disappears and solution
is colorless.
13. Read result in ppm Dissolved Oxygen.
LaMOTTE COMPANY
Helping People Solve Analytical Challenges®
PO Box 329 • Chestertown • Maryland • 21620 • USA
800-344-3100 • 410-778-3100 (Outside U.S.A.) • Fax 410-778-6394
Visit us on the web at www.lamotte.com
67414-MN • 4/07
19
Watershed Watch Project Procedures:
Collecting Samples for Laboratory Analysis
Version 3.0
2005
1
Watershed Watch
Sample Collection Methods
Table of Contents
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
Introduction
a.
Background: Watershed Watch in Kentucky
b.
Overview: Watershed Watch’s Synoptic Sampling Program
c.
Data Rigor
Training
Planning a Synoptic Sampling Event
a.
Sampling Event Coordinator
b.
Selection of Laboratory
Sampling Site Selection
a.
Objectives
b.
Maps
c.
Rationale
d.
Accessibility and Appropriateness
e.
Health and Safety
Location and Description of Sampling Sites
a.
Diagram Depicting Physical Setting
b.
Coordinates
c.
Photographs
Sample Materials
a.
Containers and Preservatives
b.
Preprinted Forms and Instructions
Sampling Procedures
a.
Completing the “Chain of Custody” form
b.
In-stream sampling Location and Approach
c.
Sampling for Parameters other than Bacteria
d.
Sampling for bacteria
e.
Sample Preservation
f.
Chemical Treatment
Transport of Samples to the Laboratory
a.
Documenting changes in custody of the sample
b.
Drop-Off Centers
c.
Runners
Laboratory Analysis
Data Management
Quality Control and Assurance
a.
How Watershed Watch Assures the Quality of Its Data
b.
Duplicate Samples and Field Blanks
2
1.
Introduction
This document describes standard operating procedures of Watershed Watch in Kentucky for
deriving reliable data about water quality samples obtained from streams for laboratory analysis
by its Volunteer Monitor participants. This section of the document will provide background on
the program, explain its organization, and introduce its sampling programs.
a.
Background: Watershed Watch in Kentucky
Watershed Watch in Kentucky educates citizens in watershed and stream science and trains them
to gather scientific data about streams.
Watershed Watch has these statewide program objectives:
• Provide citizens with an array of scientific data and an understanding of stream science
that helps them better appreciate the quality status of a stream for which they are
concerned;
• Create an annual synopsis of the overall quality of surface waters on a state and basin
basis; and
• Involve citizens who are knowledgeable about water quality in watershed planning,
protection, and restoration activities.
b.
Overview: Watershed Watch’s Synoptic Sampling Program
A major purpose of the Synoptic Assessment program of Watershed Watch is to generate data
that volunteer monitors may use to assess conditions of the stream that most interests them.
Watershed Watch trains volunteer monitors to collect the following data streamside:
• Water Quality characteristics measured at the stream including dissolved oxygen, pH,
and temperature and, in places, specific conductance;
• Physical characterization of representative stream reaches;
• Biological sampling; and
• Habitat assessment including biodiversity
This document does not discuss procedures and methods for collection of this data.
Volunteer monitors also collect grab samples about stream parameters that cannot be measured
streamside and require laboratory analysis. Samples are collected for:
• Pesticides that threaten aquatic life, sampled in the Spring;
• Human pathogens (including bacteria), sampled in the Summer; and
• Major cations and anions, trace constituents, total organic carbon and other water-quality
parameters, sampled in the Fall.
Methods and procedures for training volunteers to grab and transport samples, and for laboratory
analysis and QA/AC, are the subject of this document.
3
c.
Data Rigor
The KDOW has issued “Agency Guidance for Volunteer Monitoring Data and Reports” that indicates the
level of procedural rigor necessary for data, depending on the intended use for the data. The guidance is
summarized in the following matrix, which indicates the data-related activities that are necessary
depending on the use to which volunteer-generated data will be put:
Tier
I
II
III
IV
Data Use
Incident
Reporting
Education
Programs
Local
Awareness
Watershed
Screening
Local Planning
Activities
Effectiveness
Monitoring
TMDL
Monitoring
Use Support
Determination
X = Required
O = Optional
Consultation
with KDOW
Potential Data Rigor Requirements
SOP and
Compliance
Use of
Written
QAPP prewith federal
KDOW
Study Plan
approved
standards
SOP
by KDOW
Samplers
preapproved by
KDOW
X
X
X
X
X
O
X
X
X
X
X
X
X
O
O
X
X
X
X
X
O
X
X
X
X
X
X
KDOW = Kentucky Division of Water
QAPP = Quality Assurance Project Plan
SOP = Standard Operating Procedure
TMDL = Total Maximum Daily Load
Watershed Watch is designed to meet the data rigor of Tier II.
2.
Training
a.
Standard Sampling Curriculum
Volunteer Monitors who grab samples or supervise the sampling streamside are required to
complete a Standard Sampling Training Module developed by the Training Committee and
approved by the Science Advisors Committee of the ICC that addresses:
• Sample container handling
• Sample collection
• Sample preservation
• Sample transport and storage
• Documentation and chain of custody record completion
• QA/QC procedures including duplicate samples and field blanks
• Communication with Event Coordinators and lab staff.
The module includes a demonstration, ideally streamside, of sample container handling,
collection, and preservation, and requires the volunteer to demonstrate competency.
4
3.
Planning a "Synoptic" Sampling Event
a.
Sampling Event Coordinator
For each synoptic sampling event, the Basin Steering Committee identifies a Sampling Event
Coordinator who communicates with the receiving laboratory concerning:
o Arrangements for receiving samples (see Section 8);
o Standards for analysis (see Section 9); and
o Standard reporting spreadsheet, including flags for samples outside of standard
receiving temperature and holding time (“lab report”) (see Section 10)
The Sampling Event Coordinator also:
• Assembles packets of containers and corresponding instructions to Supervising Samplers;
• Makes arrangements for Drop-Off Centers and Runners as indicated;
b.
Selection of laboratory
Watershed Watch uses laboratories that meet at least one of the following criteria:
• Listed on the KDOW Certified Drinking Water Lab List;
• Currently providing contract work for KDOW; or
• Approved by EPPC microbiological staff.
4.
Sampling Site Selection
a.
Objectives
The site selection process in Watershed Watch attempts to accommodate its two major purposes:
• The interest of the Volunteer Monitor, who often desires to focus learning activities on a
stream reach where she or he lives, works, learns, or plays; and
• The need of the program and its stakeholders to collect information from a stream reach
near the bottom of a watershed, where data will be most representative of the condition of
the watershed.
5
b.
Site Selection Maps
The following maps are used during training to assist Volunteer Monitors and Trainers with Site
Selection:
• Kentucky Atlas and Gazetteer, ISBN Number 0-89933-216-1
• Topozone: http://www.topozone.com (Set to DD.DDDD coordinates)
• Arcview Shape Files including Counties, Roads and Streams with current Watershed
Watch Site List.
c.
Rationale
Because site selection in Watershed Watch attempts to balance the interest of the Volunteer
Monitor in a particular stream reach with the program’s objective of collecting information about
watershed conditions, site selection occurs as part of Volunteer Monitor training so that trainers
can encourage participants to:
• Join a team at an existing site;
• Open a new site at a location that will represent the condition of an undocumented
watershed; or
• Choose a stream reach that will represent the condition of their stream of interest.
d.
Accessibility and Appropriateness
The site selection unit in Watershed Watch training identifies the following factors as important in site
selection:
• Proximity to existing Watershed Watch sites;
• Access to the site using public rights of way and/or the permission of the property owner;
• Physical safety in accessing the stream via the streambank;
• Wadeability of the stream;
• Representativeness of the stream reach (channel morphology and riffles)
• Reach mixing (sites near major tributaries or point sources should be avoided to minimize
backwater effects or poorly mixed flows);
• Proximity to major man-made disturbances like bridges or dams; and
• Known health risks in the stream, e.g., proximity to a treatment plant or “straight pipes.”
e.
Health and Safety
The following health and safety factors are addressed in the training:
• Notifying others of itinerary and whereabouts;
• Never visiting an isolated site alone;
• Never sampling in high water;
• Bewaring of hunters, poisonous reptiles, and sudden high water;
• Carrying identification;
• Taking a cellular phone when available; and
• Wearing disposable, powderless gloves when handling sample preservatives such as acid.
6
5.
Location and Description of Sampling Sites
a.
Diagram depicting physical setting
The Volunteer Monitor documents the physical setting of the site using a standard form,
“Physical Characterization/ Water Quality Field Data Sheet,” which may be found in Appendix
D.
b.
Coordinates
Latitude and longitude are determined for the site in one of two ways:
• The Volunteer Monitor determines them using a handheld GPS unit and submits the
coordinates with the “Physical Characterization” form. In this instance, the GPS unit
must be tuned to the coordinate specifications called for in http://kywater.org/dow/gps/;
or
• A copy of the 1:24,000 topographic map for the stream reach is mailed to the Volunteer
Monitor with the first sampling container for the site with a request that the monitor
identify the site on the map and return it to the Data Manager.
c.
Photographs
The Volunteer Monitor is requested to submit two photographs of the stream reach with the
“Physical Characterization” form:
• Upstream of the sample point looking downstream at the sample point (marked
“downstream:); and
• Downstream of the sample point looking upstream at the sample point (marked
“upstream”).
6.
Preparation for Sampling
a.
Containers and Preservatives
Containers and preservatives for samples are obtained by Steering Committees through the ICC
purchasing cooperative or independently and meet the criteria found in Appendix E,
“Environmental Sample Preservation and Holding Times.”
The container is pre-marked with the unique Site Number by the Sampling Event Coordinator
designated by the Steering Committee before it is mailed to the Supervising Sampler for the site.
A “Chain of Custody” form (see Appendix F for the template) is prepared for each site and
preprinted with the Site Number, usually by the Data Manager. It is enclosed by the Sampling
Event Coordinator with a set of instructions. Samplers should open and read their packet upon
receipt.
7
7.
Sampling Procedures
The purposes of the following streamside sampling procedures are to assure that the sample
container correlates with the documented sampling activity and to prevent water samples from
contamination during the sampling process.
a.
Completing the “Chain of Custody” form
The Watershed Watch Chain of Custody form (Appendix F) serves to document and record the
transfer of the samples from the stream to the laboratory, functions as a field measurement form,
and provides a place for field observations. Listed below are data elements of the form:
Sample identification: the following information is preprinted on the form by the Steering
Committee’s Data Manager:
• Site Number
• Stream Name
• Watershed Number
• Sampling Location
• County
• Name of and contact information for Supervising Sampler
The following identifying information is entered by the Supervising Sampler streamside when
the sample is taken:
• Date and Time of sample collection
• Corrections to any preprinted information
Field measurements: The following information is entered by the Supervising Sampler when
and where the sample is taken:
• Comments on general stream conditions
• Flow
• Flow Rate
• Rain in past 48 hours
• Dissolved oxygen
• pH
• Temperature
• Conductivity
Signatures: The Supervising Sampler signs the form at the time she or he relinquishes it to the
laboratory or to the next person who will have custody of the sample as it is transported to the
laboratory. Signatures are annotated by the date and time they are signed.
8
b.
In-stream sampling location and approach
Samples are taken by wading to reduce sample contamination. A maximum safe wading depth
depends on the size of the person sampling, the stream’s velocity and depth, and the streambed
material. Caution should always be used when wading streams deeper than three feet.
Additional caution should be used when the streambed is composed of loose or slippery material.
Algae-coated cobbles can be slippery and as dangerous as ice. A personal floatation device
should be worn when wading streams three feet or greater in depth.
The sampler approaches the sampling site from a downstream location, walking upstream to the
sampling site, to avoid disturbing bottom sediments that could contaminate the water quality
sample. An ideal wading location is in the center of the stream and at the head of a riffle so that
water current produces a good flow past the sampling point.
c.
Sampling for parameters other than bacteria
The sampler first contaminates gloves, if worn, with stream water. Sample bottles are then
contaminated with stream water. Pre-marked sample bottles are rinsed once with stream water.
The sample bottle is then lowered from the surface to the bottom of the stream until the sample
bottle touches the stream bottom, without disturbing sediments. Upon reaching the bottom, the
bottle is raised to the surface, matching the transit rate when the bottom was lowered. Repeat
until the bottle is filled with stream water. Rinse the bottle cap in the stream and cap the bottle.
d.
Sampling for bacteria
The sampler first contaminates gloves, if worn, with stream water before sampling with the
container. Do not pre-rinse the container, and avoid contaminating the inside of the container,
especially with an ungloved finger. Dip the pre-marked sample container to a depth of about
four inches with the open end facing upstream. Push the mouth of the container upstream at this
depth until the container is nearly full. The mouth of the container should at all times be
upstream of the sample collector and any disturbed sediments. Leave enough airspace in the top
of the sample container so the sample can be remixed just before filtration at the laboratory.
Immediately chill the sample in an ice slurry (see following section).
e.
Sample preservation
Sample preservation procedures prevent reduction or loss of water quality variables of interest.
Variable loss can occur between sample collection and laboratory analysis because of physical,
chemical, and biological processes that result in chemical precipitation, absorption, oxidation,
reduction, ion exchange, degassing, or degradation. Preservation stabilizes variable
concentrations for a limited period of time. Some samples, particularly of bacteria, have a very
short holding time before laboratory analysis may begin.
In all Watershed Watch sampling events, sample containers are placed in a container with a
slurry of chilled water and ice immediately following sample collection to maintain them at 4
degrees Centigrade plus or minus 2 degrees without freezing until analyzed.
9
Sample preservation instructions are included with the sample bottles mailed or delivered to the
Supervising Sampler prior to each sampling event.
f.
Chemical treatment
If a sample requiring acidification/ chemical treatment will not be delivered to the laboratory
within six hours of its collection from the stream, the following procedures are required:
Glass ampoules containing the preservative and appropriately protected for shipping are
distributed by the Sampling Event Coordinator with the sampling containers and instructions to
Supervising Samplers. Instructions are sent that include these precautions:
“Preservatives in the glass ampoules are highly concentrated acids that must be handled
carefully. Even a small drop of the solution can burn your skin. Use of latex gloves and
safety glasses is highly recommended. Rinse each ampoule with water several times
before discarding.”
The following instructions are given:
1. After filling the container, carefully snap the neck of the ampoule and add it to
container.
2. Label the container with the letter “N” for Nitric Acid, or “S” for Sulfuric Acid, as
appropriate.
3. Place the container on ice.
4. Carefully rinse the empty preservative ampoules before discarding.
8.
Transport of Samples to the Laboratory
Chilled samples should be delivered to the laboratory as soon as possible; bacteria samples, and
samples that require acidification but have not been treated, must be delivered to the laboratory
within six hours from the time of collection.
a.
Documenting changes in custody of the sample
When the Supervising Sampler takes a sample directly to the laboratory, she or he signs, times,
and dates the Chain of Custody form in the left column when custody is relinquished to the
laboratory. The staff member of the laboratory who receives the sample similarly signs, times,
and dates the form opposite the signature of the person relinquishing it. Identical times and dates
on the same line means the sample changed custody without an intermediate step, which would
disqualify the sample from regulatory use.
b.
Drop-Off Centers
Because of the vast number of Watershed sites sampled on the same day and the few number of
receiving laboratories, “Drop-Off Centers” may be established if these criteria are met:
• The instructions that accompany sample bottles identify the person responsible for the
drop-off center, and provide directions, contact telephone number, and specific times of
operation when the responsible person will be available to accept samples; and
10
•
The center has sufficient refrigeration/cooler space to immediately chill samples
transferred from Volunteer Monitors’ coolers.
c.
Runners
Runners may be designated to collect samples from Drop-Off Centers or Volunteer Monitors in
the field. Sample runners are responsible for:
• Communicating with volunteers and drop-off locations on their route prior to sampling in
order to coordinate swift collection and transfer;
• Having sufficient cooler space to immediately chill samples transferred from Volunteer
Monitors’ coolers;
• Communicating with the destination laboratory so it is prepared to accept the samples
delivered;
• Fully understanding the delivery times required for samples; and
• Confirming that the numbering of sample containers corresponds to the number on Chain
of Custody forms;
• Signing Chain of Custody Forms when receiving and relinquishing samples;
• Checking bottle caps to assure they are securely tightened (avoid over-tightening); and
• Packing samples carefully in the receiving container to prevent bottle breakage, shipping
container leakage, and sample degradation.
9.
Laboratory Analysis
Labs selected by Watershed Watch are asked to use standard methods of analyis.
10.
Data Management
The laboratory sends its results to the project data manager. The Data Manager coordinates
review of copies of the printed report by the Sampling Event Coordinator, Steering Committee
Chair and Science Advisor for errors, omissions, and suitability. Draft copies are sent to
supervising samplers for review and comment. Once approved by the Basin Steering Committee
the monitoring data is posted on the basin web site and released at the annual conference
11. Quality Control and Assurance
a.
How Watershed Watch Assures the Quality of its Data
Quality Control and Assurance (QA/QC) is the responsibility of everyone in the chain of custody
of a sample, its analysis, and the data that results.
A first level of QA/QC is compliance with procedures and methods in the Standard Sampling
Curriculum.
The second level of QA/QC is an understanding of and compliance with these Standard
Operating Procedures among everyone involved in the sampling event.
11
The third level of QA/QC is discrete procedures for analyzing quality using the data generated in
the sampling program. These procedures are the responsibility of the Laboratory’s internal QA
program and the Steering Committee’s Data Manager and Quality Assurance Officer.
A fourth and final level of QA/QC are activities at the statewide level by the Quality Assurance
Committee that include:
• Review of QA/QC reports submitted with data by Steering Committees;
• Audits of Steering Committee QA/QC activities; and
• Comparison of data with statewide and nationwide databases;
The remainder of this section outlines QA/QC procedures that apply to Tier III
and IV data uses.
b.
Duplicate Samples and Field Blanks
The Sampling Event Coordinator selects sites for duplicate samples and field blanks in
consultation with the Steering Committee’s Science Advisor and Quality Assurance Officer.
Sites selected for duplicate and blank samples should be chosen to be representative of the range
of conditions encountered and to rotate through different sampling teams. Sites expected to be at
or near method detection limits should be included. Duplicates should also be collected where
high concentrations are expected.
1.
Duplicate Samples
Watershed Watch uses concurrent duplicate samples to assess variability in sample collection,
processing, and analysis.
Supervising Samplers for the sites selected by the random selection process receive a pre-marked
duplicate sample container in addition to the pre-marked sample containers with instructions for
taking and submitting a duplicate sample. All other procedures for samples are followed for
Duplicate Samples.
2.
Field blanks
Watershed Watch uses field blanks to assess for bias from contamination of the sample during
any stage of sample collection, processing, and analysis.
Supervising Samplers for the sites selected by the random selection process receive a pre-marked
field blank container in addition to the pre-marked sample container. The Supervising Sampler
is instructed to obtain bottled water in a food store and pour the amount required for a sample
into the container pre-marked for the field blank sample at the site and time the routine sample is
taken. All other procedures for samples are followed for field blanks. An example of
instructions to Supervising Samplers for taking field blanks may be found in Appendix G.
12
Watershed Watch Chain of Custody Record
Sample #
Date sample taken
Stream Name
Sampling Location (correct or add location info if necessary)
Time sample taken
Name of "Supervising Sampler" on site when sample collected:
Lab Notes:
If name not
correct, please
enter proper name
in Comment Box
Sampler ID#
Telephone:
Flow Rate
48 Hr Rainfall "
0-Dry
1-Ponded
2-Low
3-Normal
4-Bank Full
5-Flood!
0
0.1
0.5
1.0
1.5
2.0 +
Turbidity
0-Clear
1
2
Water Chemistry
Oxygen ppm
3-Turbid Conductivity
pH SU
Temp C
General comments, questions, corrections, concerns or suggestions.
When transporting samples to the lab, it is necessary to have each person that controls the
sample to sign when they receive it AND when they relinquish it.
Relinquished by:
Time/Date
Received by:
Time/Date
This form must accompany your sample to the lab. The first signature in the
"relinquished by" column must match the "supervising Sampler's" name!
Make a copy for yourself, then send the original on its way with your sample
runner. Please correct errors on the pre-printed part of this record. If you
have questions or difficulties, please contact us at 1-800-928-0045 Ext 473
04/2011
KGS D515
Total Phosphorus in Water
1. Discussion
MDL= 0.02 as of 5/2002
Principle
Separation into total dissolved and total recoverable forms of phosphorus depends on filtration of
the water sample through a 0.45 μm membrane filter. Total recoverable phosphorus includes all
phosphorus forms when the unfiltered, shaken sample is heated in the presence of sulfuric acid
and ammonium peroxydisulfate. Total dissolved phosphorus includes all phosphorus forms when
the filtered, shaken sample is heated in the presence of sulfuric acid and ammonium
peroxydisulfate. Phosphorus is converted to orthophosphate by digesting the water sample with
ammonium persulfate and diluted sulfuric acid. Ammonium molybdate and antimony potassium
tartrate can then react in an acid medium with dilute solutions of orthophosphate to form an
antimony-phosphate-molybdate complex. This complex is reduced to an intensely blue-colored
complex by ascorbic acid. The color intensity is proportional to the phosphorus concentration.
Sensitivity
The range of determination for this method is 0.05 mg/L to 1.00 mg/L P.
Interferences
Ferric iron must exceed 50 mg/L, copper 10 mg/L, or silica 10 mg/L, before causing an
interference. Higher silica concentrations cause positive interferences over the range of the test,
as follows: results are high by 0.005 mg/L of phosphorus for 20 mg/L of SiO2, 0.015 mg/L of
phosphorus for 50 mg/L, and 0.025 mg/L of phosphorus for 100 mg/L. Because arsenic and
phosphorus are analyzed similarly, arsenic can cause an interference if its concentration is higher
than that of phosphorus.
Sample Handling and Preparation
Samples should be preserved only by refrigeration at 4 °C. A raw sample should be used in the
analysis. The holding time for this analysis is 28 days.
2. Safety
Safety glasses, gloves, and a lab coat should be worn while performing this analysis due to the use
of, and possible exposure to, strong acids and bases.
3. Apparatus
Varion 50 Spectroscopy system
Filtration Apparatus
Coors 60242 Büchner funnels.
Suction flasks, connected in series to a vacuum system.
Reservoir for the filtrate, 500 mL.
Trap which prevents liquid from entering the vacuum system, 1000 mL
Paper filters—7.5 cm, 1 μm. (VWR Cat. # 28321-005)
Analytical balance, capable of weighing to the nearest 0.0001 g.
Drying oven.
Desiccator.
Thermix Stirring Hot Plate—Model 610T
HCl Acid washed glassware—Refer to the “Total P” section of the Glassware GLP for further
details. Commercial detergents should never used. Glassware should be dedicated for
Total P use only.
6 ½ oz. Disposable polystyrene specimen cups—Cups should be rinsed three times with DI water.
4. Reagents
Purity of Reagents—Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, all reagents shall conform to the specifications of the Committee on Analytical
Reagents of the American Chemical Society. Other grades may be used, provided it is
first ascertained that the reagent is sufficiently high in purity to permit its use without
lessening the accuracy of the determinations.
Purity of Water—Unless otherwise indicated, references to water shall be understood to mean
Type I reagent grade water (Milli Q Water System) conforming to the requirements in
ASTM Specification D1193.
Ammonium Peroxydisulfate—Place 20 g of ammonium peroxydisulfate in a 50 mL volumetric
flask. Dilute with water to volume. Add a magnetic stirrer to the flask and let the
solution stir until all the crystals have dissolved (minimum of 20 minutes). Prepare
daily.
( enough for 30 beakers total )
Solution Mixture—Dissolve 0.13 g of antimony potassium tartrate and 5.6 g of ammonium
molybdate in approximately 700 mL of water. Cautiously add 70 mL of concentrated
sulfuric acid. Allow the solution to cool and dilute to 1 liter. The solution must be kept
in a polyethylene bottle away from heat. This solution is stable for one year.
Combined Reagent—Dissolve 0.50 g solid ascorbic acid in 100 mL of solution mixture. Prepare
daily.
Phenolphthalein indicator solution—Dissolve 0.5 g of phenolphthalein in a mixture of 50 mL
isopropyl alcohol and 50 mL water.
Sulfuric acid (31 + 69)—Slowly add 310 mL of concentrated H2SO4 to approximately 600 mL of
water. Allow solution to cool and dilute to 1 liter.
Sodium Hydroxide, 10 N—Dissolve 400 g of NaOH in approximately 800 mL of water. Allow
solution to cool and dilute to 1 liter.
Sodium Hydroxide, 1 N—Dissolve 40 g of NaOH in approximately 800 mL of water. Allow
solution
to cool and dilute to 1 liter.
Phosphorus stock solution (50 mg/L)—Dissolve 0.2197 g of predried (105 °C for one hour)
KH2PO4 in water and dilute to 1 liter. Prepare daily.
Phosphorus standard solution (2.5 mg/L)—Dilute 50 mL of the stock solution to exactly 1 liter of
water. Prepare daily.
Blank—reagent grade water.
Total phosphorus stock QC solution—Using a commercially available Quality Control solution,
dilute to desired range and record manufactures name, lot #, and date.
Quality control sample—Dilute total P stock solution so that QC value falls midway in analysis
working range (0.05-1.00 ppm). Using 6.11 ppm QC stock solution, dilute 25 mL of
Total Phosphorous stock solution to 500 mL resulting in a concentration of 0.306 ppm.
Acid for glassware-Carefully add 250 mL of concentrated hydrochloric acid to approximately 600
ml of water. Dilute to 1 liter.
5. Procedure
A.
Prepare the spectrophotometer by turning on the lamp and allowing it to warm up for at least
one hour. See the Spectrophotometer GLP for a detailed listing of necessary computer
commands.
B.
Standards Prep
1. Prepare a series of phosphorus standards from the 2.5 mg/L phosphorus standard
solution according to the following table. Dilute each to 50 mL with water.
Volume of phosphorus standard, mL
1
2
4
7
10
15
20
2. Prepare all standards daily.
C.
Standard concentration, ppm
0.05
0.10
0.20
0.35
0.50
0.75
1.00
Sample Prep
1. Pour 50 mL of each of the two blanks, standards, samples, duplicates, and Total P QC’s
into 100 mL glass beakers. Add 3 - 6 glass boiling beads to each beaker.
2. Mark beakers at top of liquid with a Sharpie.
3. Add 1 mL of ammonium peroxydisulfate solution and 1 mL of H2SO4 (31+69) to each
marked beaker.
4. Place beakers on the large hot plates that are located in the hood.
5. Turn the Temp. knob on the hot plates to “HI.”
6. Let each sample (blank, standard, duplicate, or QC) stay on the hot plate until its volume
decreases to 10 mL. This process takes approximately 1 to 1 ½ hours. Do not allow the
samples to completely evaporate.
7. Allow each sample to cool in the hood.
8. Add a drop of phenolphthalein indicator solution to each sample.
9. Add 1 mL of 10 N NaOH to each sample.
10. Continue adjusting the pH’s by adding 1 N NaOH until each sample becomes faint pink
in color. The pH is approximately 10 at this point.
11. Bring samples back to colorless by adding 1 N H2SO4 to each sample. The pH is
approximately 4 at this point.
12. Bring each sample’s volume back up to the mark with water.
13. Filter each of the samples using the acid washed ceramic funnels and 1 μm paper filters.
14. Pour 25 mL of each sample into its corresponding 4 ½ oz. plastic beaker.
15. Add 5 mL of combined reagent to the sample and mix thoroughly.
16. After a minimum of 10 minutes, but no longer than 30 minutes, measure the absorbance
of the blue color at 880 nm with the spectrophotometer.
D.
Sample Analysis
1. The computer, by comparing the concentration of each calibration standard against its
absorbance, can plot a calibration curve. The correlation coefficient must be > 0.994 to
be acceptable. If above criteria is not met the standards may need to be remade and
rerun.
2. Once the spectrophotometer is standardized properly, the samples may be analyzed.
3. Once the analysis is completed, print out a copy of the standard values, plotted curve, and
the sample values. Copy the relevant data onto the Total Phosphorous Data Sheet.
E.
Clean Up
1. Turn off the spectrophotometer lamp.
2. The waste must be placed in the acid waste container.
3. For glassware clean up, refer to the “Total P” section of the Glassware GLP.
6. Quality Control
A quality control sample should be run at the beginning and end of each sample
delivery group (SDG) or at the frequency of one per every ten samples. The QC’s value should
fall between ± 10 % of its theoretical concentration.
A duplicate analysis should be run for each SDG or at the frequency of one per every twenty
samples, whichever is greater. The RPD (Relative Percent Difference) should be less than 10%.
If this difference is exceeded, the duplicate must be reanalyzed.
From each pair of duplicate analytes (X1 and X2), calculate their RPD value:
⎛ X1 − X 2 ⎞
% RPD = 2 • ⎜
⎟ x 100
⎝ X1 + X 2 ⎠
where:
(X1 - X2) means the absolute difference between X1 and X2.
7. Method Performance
The method detection limit (MDL) should be established by determining seven replicates that are
2 to 5 times the instrument detection limit. The MDL is defined as the minimum concentration
that can be measured and reported with 99% confidence that the analyte concentration is greater
than zero and is determined from analysis of a sample in a given matrix containing the analyte.
MDL = t ( n −1,1−α = 99 ) ( S )
where:
t = the t statistic for n number of replicates used (for n=7, t=3.143)
n = number of replicates
S = standard deviation of replicates
8. References
ASTM vol. 11.01 (1996), D 515, “Standard Test Methods for Phosphorus in Water”, pg. 24
ASTM vol. 11.01 (1996), D 1193, “ Specification for Water”, pg. 116
EPA 365.2
Phosphorous , All Forms (Colorimetric, Ascorbic Acid)
01/2009
KGS 9056
Ion Chromatography of Water
1. Discussion
Principle
This method addresses the sequential determination of the following inorganic anions: bromide,
chloride, fluoride, nitrate, Kjeldahl nitrogen, total nitrogen and sulfate. A small volume of water
sample is injected into an ion chromatograph to flush and fill a constant volume sample loop. The
sample is then injected into a stream of carbonate-bicarbonate eluent. The sample is pumped
through three different ion exchange columns and into a conductivity detector. The first two
columns, a precolumn (or guard column), and a separator column, are packed with low-capacity,
strongly basic anion exchanger. Ions are separated into discrete bands based on their affinity for
the exchange sites of the resin. The last column is a suppressor column that reduces the
background conductivity of the eluent to a low or negligible level and converts the anions in the
sample to their corresponding acids. The separated anions in their acid form are measured using
an electrical conductivity cell. Anions are identified based on their retention times compared to
known standards. Quantitation is accomplished by measuring the peak area and comparing it to a
calibration curve generated from known standards.
Sensitivity
Ion Chromatography values for anions ranging from 0 to approximately 40 mg/L can be measured
and greater concentrations of anions can be determined with the appropriate dilution of sample
with deionized water to place the sample concentration within the working range of the calibration
curve.
Interferences
Any species with retention time similar to that of the desired ion will interfere. Large quantities of
ions eluting close to the ion of interest will also result in interference. Separation can be improved
by adjusting the eluent concentration and /or flow rate. Sample dilution and/or the use of the
method of Standard Additions can also be used. For example, high levels of organic acids may be
present in industrial wastes, which may interfere with inorganic anion analysis. Two common
species, formate and acetate, elute between fluoride and chloride. The water dip, or negative
peak, that elutes near, and can interfere with, the fluoride peak can usually be eliminated by the
addition of the equivalent of 1 mL of concentrated eluent (100X) to 100 mL of each standard and
sample. Alternatively, 0.05 mL of 100X eluent can be added to 5 mL of each standard and
sample.
Because bromide and nitrate elute very close together, they can potentially interfere with each
other. It is advisable not to have Br-/NO3- ratios higher than 1:10 or 10:1 if both anions are to be
quantified. If nitrate is observed to be an interference with bromide, use of an alternate detector
(e.g., electrochemical detector) is recommended.
Method Interferences may be caused by contaminants in the reagent water, reagents, glassware,
and other sample processing apparatus that lead to discrete artifacts or elevated baseline in ion
chromatograms. Samples that contain particles larger than 0.45 micrometers and reagent solutions
that contain particles larger than 0.20 micrometers require filtration to prevent damage to
instrument columns and flow systems. If a packed bed suppressor column is used, it will be slowly
consumed during analysis and, therefore, will need to be regenerated. Use of either an anion fiber
suppressor or an anion micro-membrane suppressor eliminates the time-consuming regeneration
step by using a continuous flow of regenerant.
Because of the possibility of contamination, do not allow the nitrogen cylinder to run until it is
empty. Once the regulator gauge reads 100 kPa, switch the cylinder out for a full one. The old
cylinder should them be returned to room #19 for storage until the gas company can pick it up.
Make sure that the status tag marks the cylinder as “EMPTY”.
Sample Handling and Preservation
Samples should be collected in glass or plastic bottles that have been thoroughly cleaned and
rinsed with reagent water. The volume collected should be sufficient to ensure a representative
sample and allow for replicate analysis, if required. Most analytes have a 28 day holding time,
with no preservative and cooled to 4oC. Nitrite, nitrate, and orthophosphate have a holding time
of 48 hours. Combined nitrate/nitrite samples preserved with H2SO4 to a pH <2 can be held for 28
days; however, pH<2 and pH>12 can be harmful to the columns. It is recommended that the pH
be adjusted to pH>2 and pH<12 just prior to analysis.
Note: Prior to analysis, the refrigerated samples should be allowed to equilibrate
to room temperature for a stable analysis.
2. Apparatus
Dionex DX500
Dionex CD20 Conductivity Detector
Dionex GP50 Gradient Pump
Dionex Eluent Organizer
Dionex AS40 Automated Sampler
Dionex ASRS-Ultra Self-Regenerating Suppressor
Dionex Ionpac Guard Column (AG4A, AG9A, or AG14A)
Dionex Ionpac Analytical Column (AS4A, AS9A, or AS14A)
Dionex Chromeleon 6.8 Software Package
Dionex 5 mL Sample Polyvials and Filter Caps
2 L Regenerant Bottles
5 mL Adjustable Pipettor and Pipettor Tips
1 mL Adjustable Pipettor and Pipettor Tips
A Supply of Volumetric Flasks ranging in size from 25 mL to 2 L
A Supply of 45 micrometer pore size Cellulose Acetate Filtration Membranes
A Supply of 25x150 mm Test Tubes
Test Tube Racks for the above 25x150 mm Test Tubes
Gelman 47 mm Magnetic Vacuum Filter Funnel, 500 mL Vacuum Flask, and a Vacuum Supply
3. Reagents
Purity of Reagents—HPLC grade chemicals (where available) shall be used in all reagents for Ion
Chromatography, due to the vulnerability of the resin in the columns to organic and trace
metal contamination of active sites. The use of lesser purity chemicals will degrade the
columns.
Purity of Water—Unless otherwise indicated, references to water shall be understood to mean
Type I reagent grade water (Milli Q Water System) conforming to the requirements in
ASTM Specification D1193.
Eluent Preparation for SYSTEM2 NITRATE Methods, including Bromides (using AG4, AG4
and AS4 columns)—All chemicals are predried at 105° C for 2 hrs then stored in the
desiccator. Weigh out 0.191 g of sodium carbonate (Na2CO3) and 0.286 g of sodium
bicarbonate (NaHCO3) and dissolve in water. System 2 (the chromatography module that
contains the AG4, AG4, and AS4 Dionex columns) to be sparged, using helium, of all
dissolved gases before operation.
Eluent Preparation for SYSTEM2 NITRATE (F) Method (using AG14 and AS14 columns)—
Weigh out 0.3696 g of sodium carbonate (Na2CO3) and 0.080 g of sodium bicarbonate
(NaHCO3) and dissolve in water. Bring the volume to 1000 mL and place the eluent in
the System 1 bottle marked for this eluent concentration. The eluent must be sparged
using helium as in the above reagent for System 2.
Eluent Preparation for SYSTEM2 TKN (TKN) Methods, including Total Nitrogen (using AG4A,
AG4A, and AS4A columns)—Weigh out 0.191 g of sodium carbonate (Na2CO3) and
0.143 g of sodium bicarbonate (NaHCO3) and dissolve in water. Bring the volume up to
1000 ml and place in the System 2 bottle labeled “IC-TKN 0.191/0.143”. Sparge the
eluent as in the above reagent for System 2.
100X Sample Spiking Eluent—prepared by using the above carbonate/bicarbonate ratios, but
increasing the concentration 100X. Weigh out 1.91 g of Na2CO3 and 2.86 g of NaHCO3
into a 100 mL volumetric flask. 0.05 mL of this solution is added to 5 mL of all samples
and standards to resolve the water dip associated with the fluoride peak.
Stock standard solutions, 1000 mg/L (1 mg/mL): Stock standard solutions may be purchased
(SPEX) as certified solutions or prepared from ACS reagent grade materials (dried at
105o C for 30 minutes
Calibration Standards—for the SYSTEM2 NITRATE (except Bromide) methods are prepared as
follows:
1. Calibration Standard 1: Pipette 0.1 mL of 1000 mg/L NaNO3 stock standard, 0.1 mL of
1000 mg/L NaF stock standard, 2 mL of 1000 mg/L NaCl stock standard, and 10 mL of
1000 mg/L K2SO4 stock standard into a 1000 mL volumetric flask partially filled with
water, then fill to volume.
2. Calibration Standard 2: Pipette 0.5 mL of 1000 mg/L NaNO3 stock standard, 0.5 mL of
1000 mg/L NaF stock standard, 5 ml of 1000 mg/L NaCl stock standard, and 20 mL of
1000 mg/L K2SO4 stock standard into a 1000 mL volumetric flask, partially filled with
water, then fill to volume.
3. Calibration Standard 3: Pipette 2.5 mL of 1000 mg/mL NaNO3 stock standard, 2.5 mL of
1000 mg/L NaF stock standard, 10 mL of 1000 mg/L NaCl stock standard, and 40 mL of
1000 mg/L K2SO4 stock standard into a 1000 mL volumetric flask partially filled with
deionized water, then fill to volume.
4. Quality Control Sample: Pipette 1.0 mL of 1000 mg/L NaNO3 stock solution, 1.0 mL of
1000 mg/L NaF stock solution, 8 mL of 1000 mg/L NaCl stock solution, and 30 mL of
mg/L K2SO4 stock standard into a 1000 mL volumetric flask, partially filled with water,
then fill to volume.
Calibration Standards—for the SYSTEM2 NITRATE (Fluoride) method are prepared as
follows:
1. Calibration Standard 1: Pipette 0.01 mL of 1000 mg/L NaF stock standard into a 1000
mL volumetric flask partially filled with water, then fill to volume.
2. Calibration Standard 2: Pipette 0.05 mL of 1000 mg/L NaF stock standard into a 1000
mL volumetric flask partially filled with water, then fill to volume.
3.
4.
5.
6.
7.
8.
Calibration Standard 3: Pipette 0.1 mL of 1000 mg/mL NaF stock standard into a 1000
mL volumetric flask partially filled with water, then fill to volume.
Calibration Standard 4: Pipette 0.5 mL of 1000 μg/mL NaF stock standard into a 1000
mL volumetric flask partially filled with water, then fill to volume.
Calibration Standard 5: Pipette 1.0 mL of 1000 mg/L 1000 stock standard into a 1000 mL
volumetric flask partially filled with water, then fill to volume.
Quality Control Standard: Pipette 0.1 mL of 1000 mg/L NaF from a separate source stock
standard into a 1000 mL volumetric flask partially filled with water, then fill to volume.
Quality Control Standard: Pipette 0.4 mL of 1000 mg/L NaF from a separate source stock
standard into a 1000 mL volumetric flask partially filled with water, then fill to volume.
Quality Control Standard: Pipette 1.0 mL of 1000 mg/L NaF from a separate source stock
standard into a 1000 mL volumetric flask partially filled with water, then fill to volum
Calibration Standards—for the SYSTEM2 NITRATE (Bromide) method are prepared as
follows:
1.
2.
3.
4.
Calibration Standard 1: Pipette 2 mL of 1000 mg/L NaBr stock standard into a 1000 mL
volumetric flask partially filled with water, then fill to volume.
Calibration Standard 2: Pipette 5 mL of 1000 mg/L NaBr stock standard into a 1000 mL
volumetric flask partially filled with water, then fill to volume.
Calibration Standard 3: Pipette 10 mL of 1000 mg/L NaBr stock standard into a 1000 mL
volumetric flask partially filled with water, then fill to volume.
Quality Control Standard: Pipette 8 mL of 1000 mg/L NaBr stock standard into a 1000
mL volumetric flask partially filled with water, then fill to volume.
Outside Source Certified Quality Control Sample—ERA
4. Procedure
A.
Instrument Preparation
1. Before turning on the Dionex Ion Chromatography System:
a. Fill the eluent reservoir(s) with fresh eluent.
b. Make certain the waste reservoir is empty of all waste.
c. Turn on the helium. The system pressure should be between 7 - 15psi. The system
pressure can be regulated with the knob on the back of the Eluent Organizer.
d. Connecting a piece of tubing to the gas line going into the eluent bottle and putting
the tubing into the eluent degasses the eluent reservoir(s). The gas knob on the
Eluent Organizer that corresponds to the eluent bottle should be slowly opened until
a constant bubbling stream can be seen in the eluent bottle.
e. The eluent should be degassed with helium, for a minimum of 30 minutes, before
operation of the instrument.
f. After the eluent has been degassed, remove the tube from the eluent and tightly seal
the eluent bottle. The eluent is now ready to introduce into the system.
2. Whether using the IP25 for Fluorides or the GP50 for everything else, turn off the
browser, scroll to REMOTE on the screen, select LOCAL and ENTER.
3. Scroll to mL/min., change to 0 mL/min., and hit ENTER. If using the IP25 pump, skip
to step #5.
4. Hit MENU and select 1, then ENTER.
5. Insert syringe into the Priming Block, open the gas valve on the Eluent Organizer, turn
the valve on the Priming Block counterclockwise, and turn on the pump that corresponds
with the method to be ran by pushing the OFF/ON button.
6. If the syringe does not fill freely, assist by gently pulling back on the plunger of the
syringe. Make certain that all of the air bubbles are removed from the eluent line to the
pumps.
7. Press OFF/ON on the pump to turn it off.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
Turn the valve on the Priming Block clockwise, remove the syringe and expel the air
bubbles from the syringe.
Reinsert the syringe filled with eluent into the Priming Block.
Open the valve on the Pressure Transducer and the valve on the Priming Block with the
eluent filled syringe still attached. This is accomplished by turning both
counterclockwise.
Press PRIME on the pump and push the contents of the syringe into the Priming Block.
After the eluent has been injected into the Priming Block, press OFF/ON to turn the
prime pump off and to close the valves on the Pressure Transducer and Priming Block.
Remove the syringe from the Priming Block.
Scroll to the mL/min. on the screen for the pump. For the GP50, type 2 mL/min., and
press ENTER. For the IP25, type 1.2 mL/min., and press ENTER.
Press OFF/ON to turn on the pump at the appropriate rate. The pressure should soon
stabilize between both pumpheads after two minutes of pumping time.
If the pressure between pumpheads has a difference >20 psi, then shut down the pump
and repeat steps 2-14 to remove air bubbles and prime the pumps.
Once the pump has a pumping pressure difference between pumpheads of <20 psi, then
go to the computer and enter PeakNet.
On the computer, turn on the Chromeleon 6.8 browser, then choose either System 1
(Fluoride) or System 2 (all other anions including Bromide and TKN).
Go to last run sequence, click to highlight and go to file, click save as.. This will load
the method of interest and a template for the current sequence run.
The sequence is edited to reflect the method and samples that are to be run.
a. SYSTEM2 NITRATE for Fluoride
b. SYSTEM2 NITRATE for Bromides
c. SYSTEM2 TKN for TKN and Total Nitrogen
Note: Data is reprocessed in the section of Chromelon 6.8 called Sequence integration
editor. Only operators with a minimum of three months experience in Ion
Chromatography should attempt to reprocess data for this analysis. Once data is
optimized, then the nitrogen values from nitrate and nitrite analysis can be subtracted
from this value for the TKN nitrogen value. If only Total Nitrogen is needed then use
the optimized data value without the correction for nitrite and nitrate nitrogen.
d. SYSTEM 2 NITRATE for all other anions,
20. Observe the reading on the screen of the CD20 Conductivity Detector. A conductivity
rate change of <0.03 μS over a 30 second time span is considered stable for analysis.
21. If using the GP50 pump, it will take about 15-30 minutes for the CD20 system to
stabilize. If using the IP25, it will take between 30 minutes to 2 hours for stabilization.
22. Once the CD20 is stabilized, the Dionex DX500 Ion Chromatography
System is ready to start standardization.
NOTE: When using the GP50 Gradient Pump, all due care must be taken before one
switches from local procedures to remote procedures. The bottle from which the eluent
is being pumped (i.e., A, B, C, or D) must exactly match the bottle specified in the
method. If there is a difference, then once the pump control is turned over to remote
control, irreversible damage and destruction of suppressors, columns, piston seals, and
check valves on the GP50 Gradient Pump will occur. NEVER switch from bottle C to
A, B, or D without flushing the system lines with water to remove all traces of eluent
from bottle C from the lines.
B.
Sample Preparation
1. If the sample was not filtered in the field, it must be done so now. Transfer 50 mL of a
well-mixed sample to the filtering apparatus. Apply the suction and collect the filtrate.
2.
If the conductivity values for the sample are high, dilution will be necessary to properly
run the sample within the calibration standard range. Dilutions are made in the Polyvials
with the plastic Filter Caps. If the dilutions are > 20X, then volumetric glassware is
required.
3. All dilutions are performed with reagent grade DI water. Be sure to mix the dilution
well.
4. For Fluorides and Bromides, pipette 5.0 mL of the filtered samples into the Polyvials.
For all other anions, including TKN and Total Nitrogen, first pipette 0.05 mL of 100X
sample spiking eluent into the Polyvials, then pipette 4.95 mL of the filtered samples on
top of the spiking eluent.
5. The Filter Caps are pressed into the Polyvials using the insertion tool.
6. Place the Polyvials into the Sample Cassette, which is placed into the Autosampler.
7. The white/black dot on the Sample Cassette should be located on right-hand side when
loaded in the left-hand side of the Automated Sampler for System 2.
8. For every ten samples the following should be included:
a. 1 DI water blank
b. 1 Duplicate of any one sample
c. 1 Quality Control sample/calibration check
C.
Calibration and Sample Analysis
1. Set up the instrument with proper operating parameters established in the operation
condition procedure
2. The instrument must be allowed to become thermally stable before proceeding. This
usually takes 1 hour from the point on initial degassing to the stabilization of the baseline
conductivity.
3. To run samples on the Dionex Ion Chromatography System:
a. Make a run schedule on the Chromeleon 6.8 Software Section labeled SEQUENCE.
b. Double click the mouse on the SYSTEM 1 SEQUENCES or SYSYTEM 2
SEQUENCES to display the Scheduler Area. The name of the calibration standards
must be entered under the sample name section as Standard #1, Standard #2, and
Standard #3.
Note: Level must be changed to the corresponding standard level or the calibration
will be in error. (Example: Standard #1 = Level #1; Standard #5 = Level #5)
c.
d.
4.
5.
6.
7.
Next, enter QC, blanks, QC, samples, duplicates, QC, and blanks, in that order.
Under sample type, click on either Calibration Standard or Sample, depending on
what is being run.
e. Under the Method section, the method name must be entered. To do so, double
click on the highlighted area under Method, scroll through the list of methods and
double click on the method of interest.
f. Next under the Data File section, enter the name of the data file.
g. Finally, in the Dil area, type in the dilution factor if different from 1. Do this for all
standards, blanks, quality controls, duplicates, and samples to be run under this
schedule.
h. Save the schedule and obtain a printout of it.
i. Standardize the Dionex Ion Chromatography System by running the standards:
Standard #1, Standard #2, and Standard #3.
Run the QC standards.
Run the prepblank and DI water blank.
Run the samples, duplicates, and blanks.
Run the QC standards at the end.
5. Calculations
A. Calculations are based upon the ratio of the peak area and concentration of standards to the
peak area for the unknown. Peaks at the same or approximately the same retention times are
compared. Once the method has been updated with the current calibration, this is calculated
automatically by the software using linear regression. Remember that when dilutions are
being run, the correct dilution factor must be entered.
B. Manual calculations are based upon the ratio of the peak and concentration of standards to the
peak area for the unknown when the software will not automatically calculate the unknown
concentration. Peaks at the same or approximately the same retention times are compared.
The unknown concentration can be calculated from using this ratio. Remember that when
dilutions are being run that the correct dilution factor must be entered before you will get the
correct result.
C. When possible the unknown should be bracketed between two knowns and the calculation of
the unknown made from both for comparison.
6. Quality Control
A quality control sample obtained from an outside source must first be used for the initial
verification of the calibration standards. A fresh portion of this sample should be analyzed
every week to monitor stability. If the results are not within +/- 10 % of the true value listed for
the control sample, prepare a new calibration standard and recalibrate the instrument. If this does
not correct the problem, prepare a new standard and repeat the calibration. A quality control
sample should be run at the beginning and end of each sample delivery group (SDG) or at the
frequency of one per every ten samples. The QC’s value should fall between ± 10 % of its
theoretical concentration.
A duplicate should be run for each SDG or at the frequency of one per every twenty samples,
whichever is greater. The RPD (Relative Percent Difference) should be less than 10%. If this
difference is exceeded, the duplicate must be reanalyzed.
From each pair of duplicate analytes (X1 and X2), calculate their RPD value:
⎛ X1 − X 2 ⎞
% RPD = 2 • ⎜
⎟ x 100
⎝ X1 + X 2 ⎠
where:
(X1 - X2) means the absolute difference between X1 and X2.
7. Method Performance
The method detection limit (MDL) should be established by determining seven replicates that are
2 to 5 times the instrument detection limit. The MDL is defined as the minimum concentration
that can be measured and reported with 99% confidence that the analyte concentration is greater
than zero and is determined from analysis of a sample in a given matrix containing the analyte.
MDL = t ( n −1,1−α = 99 ) ( S )
where:
t = the t statistic for n number of replicates used (for n=7, t=3.143)
n = number of replicates
S = standard deviation of replicates
8. Reference
EPA SW 846-9056, Chapter 5, September 1994
U.S. EPA Method 300.0, March 1984
ASTM vol. 11.01 (1996), D 4327, “Standard Test Method for Anions in Water by
Chemically
Suppressed Ion Chromatography”.
0/2010 addendum to 01/2009 Ion Chromatography of Water
1.
Discussion
Principle
and iodine.
3.
Reagents
Calibration Standards
1. Calibration Standard 1: Pipette 0.1 mL of 1000 mg/L I stock standard into a
1000 mL volumetric flask partially filled with water, then fill to volume.
2. Calibration Standard 2: Pipette 0.5 mL of 1000 mg/L I stock standard into a
1000 mL volumetric flask partially filled with water, then fill to volume.
3. Calibration Standard 3: Pipette 1.0 mL of 1000 mg/L I stock standard into a
1000 mL volumetric flask partially filled with water, then fill to volume.
4. Calibration Standard 4: Pipette 5.0 mL of 1000 mg/L I stock standard into a
1000 mL volumetric flask partially filled with water, then fill to volume.
5. Calibration Standard 5: Pipette 10.0 mL of 1000 mg/L I stock standard into a
1000 mL volumetric flask partially filled with water, then fill to volume.
6. Quality Control Sample: Pipette 5.0 mL of 1000 mg/L I stock standard into a
1000 mL volumetric flask partially filled with water, then fill to volume.
08/2010
KGS 365.3
Orthophosphate as Phosphate in Water
1. Discussion
MDL 0.003 as of 5/2002
MDL 0.009 as PO4
Principle
Ammonium molybdate and antimony potassium tartrate react in an acid medium with dilute
solutions of orthophosphate to form an antimony-phospho-molybdate complex. This complex is
reduced to an intensely blue-colored complex by ascorbic acid. The color is proportional to the
orthophosphate concentration.
Sensitivity
The range of determination for this method is 0.010 mg/L to 1.000 mg/L (in the PO4 –P form),
when analyzed at 880.0 nm. (Method states using 650nm wavelength or 880.0nm if instrument is
capable and the Cary 50 UV-Vis is capable).
Interferences
Arsenate, iron and silica cause interferences. Reducing the arsenic acid to arsenious acid with
sodium bisulfite may eliminate arsenate interference. When high concentrations of iron are
present, recovery of phosphorus will be lowered because the iron will use some of the reducing
agent. The bisulfite treatment will also eliminate this interference. High silica concentrations
cause positive interference.
Sample Handling and Preparation
If possible, a filtered aliquot should be used. If the analysis cannot be performed the day of
collection, the sample should be preserved by the addition of 2 ml concentrated H2SO4 per liter
and refrigerated at 4 °C. Recommended holding time is 48 hours. Suspended solids should be
removed by filtration.
Note: Samples should be filtered PRIOR to preservation.
2. Safety
Safety glasses, gloves, and a lab coat should be worn while performing this analysis due to the use
of and possible exposure to strong acids.
3. Apparatus
Varion 50 Spectroscopy system
Filtration Apparatus:
Gelman 47mm magnetic filter funnel.
Suction flasks, connected in series to a vacuum system.
Reservoir for the filtrate, 500 mL.
Trap which prevents liquid from entering the vacuum system, 1000 mL.
Cellulose-acetate filters—Micron Separations, Inc. 47 mm, 0.45 micron cellulose acetate filter
membrane.
Analytical balance—capable of weighing to the nearest 0.0001 g.
Drying oven.
Desiccator.
4 1/2 oz. plastic beakers—Must be single-use only. Rinse three times with DI water.
4. Reagents
Purity of Reagents—Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, all reagents shall conform to the specifications of the Committee on Analytical
Reagents of the American Chemical Society. Other grades may be used, provided it is
first ascertained that the reagent is sufficiently high in purity to permit its use without
lessening the accuracy of the determinations.
Purity of water—Unless otherwise indicated, references to water shall be understood to mean
Type 1 reagent grade water (Milli Q Water System) conforming to the requirements in
ASTM Specification D1193.
Ammonium molybdate-antimony potassium tartrate solution—This solution has a long shelf life
and is stable for approximately six months. It is stored in a one liter plastic bottle.
Dissolve 8 g of ammonium molybdate and 0.2 g antimony potassium tartrate in 800 mL
water and dilute to 1 liter.
Ascorbic acid solution—This solution has a shelf life of two weeks and should be stored in a one
liter amber bottle (in the refrigerator). Be sure to indicate on the label when the solution
was made. Dissolve 15 g of ascorbic acid in 125 mL water and dilute to 250 mL. Add
0.5 mL of spec. grade acetone.
Sulfuric acid, 12 N—This solution is stable for up to six months. It is stored in a one liter plastic
bottle. Slowly add 333.3 ml concentrated H2SO4 to approximately 600 mLwater. Allow
solution to cool and dilute to 1000 mL.
Phosphate as P Stock (1000ppm)--Purchases stock from ERA
Phosphate as P Standard Solution (10mg/L)—5mls of 1000mg/L stock in 500ml flask dilute with
Milli-Q water
Quality control sample—Dilute ortho-p QC stock solution so that the QC value falls within
analysis working range (0.01-1.00). QC is purchased from ERA.
Ortho-P QC stock solution—Using a commercially available Quality Control solution, dilute to
desired range and record manufacturers name, lot #, and date.
Blank—Reagent Grade DI Water
5. Procedure
A. Prepare the spectrophotometer by turning on the lamp and allowing it to warm up for at least
one hour. See the Spectrophotometer GLP for a detailed listing of necessary computer
commands.
B.
Standards Prep
1. Prepare a series of phosphorus standards from the phosphorus standard solution (10
mg/L) according to the following table-dilute to volume with water.
Volume of standard, mL
0.5
1
Standard concentration, ppm
0.02
0.05
Final Volume, mL
250
200
5
5
10
20
50.0
2.
B.
0.10
0.20
0.50
0.80
1.00
500
250
200
250
500
These standards must be prepared daily.
Sample Prep
If the samples were not filtered in the field, follow below procedure to do so now.
a. Begin by rinsing all filtering apparatuses with water (magnetic filter funnel, magnetic
filter membrane, and suction flask).
b. Place the magnetic filter membrane tightly on the suction flask and turn on the
vacuum. This will remove any water remaining in the filter funnel. After a few
seconds, turn off the vacuum pump.
c. Using small tweezers, place a cellulose-acetate filter on each of the magnetic filter
membranes and turn on the vacuum pump. Place a magnetic filter funnel on top of
each magnetic filter membrane.
d. Pour a small amount of sample (10 mL or less) into the funnel and let it collect in the
suction flask. When the entire sample has drained through, stop suction; disconnect
the suction hose from one flask at the plastic connector junction.
e. Remove both the magnetic funnel and the magnetic filter membrane together (do not
disturb the filter) and carefully lower a 50 mL glass collection tube into the suction
flask. Reconnect the apparatus and filter the sample into the collection tube.
f. Once filtered, measure out 50 mL of sample and transfer it into a 100 mL plastic
beaker (measure using a graduated cylinder). This may require two separate
filterings.
2. Add 1 mL of 12 N H2SO4 and 4 mL of ammonium molybdate-antimony potassium
tartrate to all standards and sample samples. Swirl to mix thoroughly.
3. Add 2 mL of ascorbic acid solution to all standards and samples. Swirl to mix
thoroughly.
4. After waiting 5 minutes, the samples, standards, and QC’s are ready to be analyzed at 880
nm. The blue color is stable for one hour.
1.
C.
Sample Analysis
1. Place each standard and / or sample under probe submersing probe in solution swirl
gently tap probe(releasing bubbles) and read or pour into cuvet and read.
2. The computer, by comparing the concentration of each calibration standard against its
absorbance, can plot a calibration curve. The correlation coefficient must be > 0.994 to
be acceptable. If above criteria is not met, the standards may need to be remade and
rerun.
3. Once the spectrophotometer is standardized properly, the samples may be analyzed.
4. When completed, print out a copy of the standard values, plotted curve, and the sample
values. Copy the relevant data onto the Orthophosphate Data Sheet.
D.
Clean Up
1. Turn off the spectrophotometer lamp.
2. Pour waste in acidic waste container.
3. For glassware clean up, refer to the “NORMAL” section of the Glassware GLP.
6. Calculations
The value read from the spectrophotometer is in the form PO4-P. This value may need to be
converted to the PO4 form. The equation for this conversion is: PO4- P \ 0.32614 = PO4.
7. Quality Control
A calibration curve should be established using the standards described in section 3. Each point
on this curve must be within ± 10 % error or the calibration curve must have a r2 value of 0.994 or
above. Any point that is not within this range or that causes a low r2 value must be redeveloped
and rerun until either above criteria are satisfied.
The quality control sample should be run at the beginning and end of each sample delivery group
(SDG) or at the frequency of one per every ten samples. The QC’s value should fall between ± 10
% of its theoretical concentration.
A duplicate should be run for of each SDG or at the frequency of one per every twenty samples,
whichever is greater. The RPD (Relative Percent Difference) should be less than 10%. If this
difference is exceeded, the sample must be reanalyzed.
From each pair of duplicate analytes (X1 and X2), calculate their RPD value:
⎛ X1 − X 2 ⎞
% RPD = 2 • ⎜
⎟ x 100
⎝ X1 + X 2 ⎠
where:
(X1 - X2) means the absolute difference between X1 and X2.
If a sample’s value exceeds 1.000 ppm, the sample must be diluted. The sample must be diluted
so that its concentration falls between 0.010 ppm and 1.000 ppm. The sample must be diluted
using volumetric flasks and pipettes.
8. Method Performance
The method detection limit (MDL) should be established by determining seven replicates that are
2 to 5 times the instrument detection limit. The MDL is defined as the minimum concentration
that can be measured and reported with 99% confidence that the analyte concentration is greater
than zero and is determined from analysis of a sample in a given matrix containing the analyte.
MDL = t ( n −1,1−α = 99 ) ( S )
where:
t = the t statistic for n number of replicates used
n = number of replicates
S = standard deviation of replicates
9. References
U.S. EPA Method 365.3, 1978
Standard Methods for the Examination of Water and Wastewater, 20th edition (1998),
Method 4500-P E, pg. 4-146
01/2009
KGS 9056
TKN & Total Nitrogen by Ion Chromatography of Water
1. Discussion
Principle
This method addresses the sequential determination of the following inorganic anions: bromide,
chloride, fluoride, nitrate, Kjeldahl nitrogen, total nitrogen and sulfate. A small volume of water
sample is injected into an ion chromatograph to flush and fill a constant volume sample loop. The
sample is then injected into a stream of carbonate-bicarbonate eluent. The sample is pumped
through three different ion exchange columns and into a conductivity detector. The first two
columns, a precolumn (or guard column), and a separator column, are packed with low-capacity,
strongly basic anion exchanger. Ions are separated into discrete bands based on their affinity for
the exchange sites of the resin. The last column is a suppressor column that reduces the
background conductivity of the eluent to a low or negligible level and converts the anions in the
sample to their corresponding acids. The separated anions in their acid form are measured using
an electrical conductivity cell. Anions are identified based on their retention times compared to
known standards. Quantitation is accomplished by measuring the peak area and comparing it to a
calibration curve generated from known standards.
Sensitivity
Ion Chromatography values for anions ranging from 0 to approximately 40 mg/L can be measured
and greater concentrations of anions can be determined with the appropriate dilution of sample
with deionized water to place the sample concentration within the working range of the calibration
curve.
Interferences
Any species with retention time similar to that of the desired ion will interfere. Large quantities of
ions eluting close to the ion of interest will also result in interference. Separation can be improved
by adjusting the eluent concentration and /or flow rate. Sample dilution and/or the use of the
method of Standard Additions can also be used. For example, high levels of organic acids may be
present in industrial wastes, which may interfere with inorganic anion analysis. Two common
species, formate and acetate, elute between fluoride and chloride. The water dip, or negative
peak, that elutes near, and can interfere with, the fluoride peak can usually be eliminated by the
addition of the equivalent of 1 mL of concentrated eluent (100X) to 100 mL of each standard and
sample. Alternatively, 0.05 mL of 100X eluent can be added to 5 mL of each standard and
sample.
Because bromide and nitrate elute very close together, they can potentially interfere with each
other. It is advisable not to have Br-/NO3- ratios higher than 1:10 or 10:1 if both anions are to be
quantified. If nitrate is observed to be an interference with bromide, use of an alternate detector
(e.g., electrochemical detector) is recommended.
Method Interferences may be caused by contaminants in the reagent water, reagents, glassware,
and other sample processing apparatus that lead to discrete artifacts or elevated baseline in ion
chromatograms. Samples that contain particles larger than 0.45 micrometers and reagent solutions
that contain particles larger than 0.20 micrometers require filtration to prevent damage to
instrument columns and flow systems. If a packed bed suppressor column is used, it will be slowly
consumed during analysis and, therefore, will need to be regenerated. Use of either an anion fiber
suppressor or an anion micro-membrane suppressor eliminates the time-consuming regeneration
step by using a continuous flow of regenerant.
Because of the possibility of contamination, do not allow the nitrogen cylinder to run until it is
empty. Once the regulator gauge reads 100 kPa, switch the cylinder out for a full one. The old
cylinder should them be returned to room #19 for storage until the gas company can pick it up.
Make sure that the status tag marks the cylinder as “EMPTY”.
Sample Handling and Preservation
Samples should be collected in glass or plastic bottles that have been thoroughly cleaned and
rinsed with reagent water. The volume collected should be sufficient to ensure a representative
sample and allow for replicate analysis, if required. Most analytes have a 28 day holding time,
with no preservative and cooled to 4oC. Nitrite, nitrate, and orthophosphate have a holding time
of 48 hours. Combined nitrate/nitrite samples preserved with H2SO4 to a pH <2 can be held for 28
days; however, pH<2 and pH>12 can be harmful to the columns. It is recommended that the pH
be adjusted to pH>2 and pH<12 just prior to analysis. Preserved samples should not be used for
TKN and Total Nitrogen analysis.
Note: Prior to analysis, the refrigerated samples should be allowed to equilibrate
to room temperature for a stable analysis.
2. Apparatus
Dionex DX500
Dionex CD20 Conductivity Detector
Dionex GP50 Gradient Pump
Dionex Eluent Organizer
Dionex AS40 Automated Sampler
Dionex ASRS-Ultra Self-Regenerating Suppressor
Dionex Ionpac Guard Column (AG4A)
Dionex Ionpac Analytical Column (AS4A)
Dionex Chromeleon 6.8 Software Package
Dionex 5 mL Sample Polyvials and Filter Caps
2 L Regenerant Bottles
5 mL Adjustable Pipettor and Pipettor Tips
1 mL Adjustable Pipettor and Pipettor Tips
A Supply of Volumetric Flasks ranging in size from 25 mL to 2 L
A Supply of 45 micrometer pore size Cellulose Acetate Filtration Membranes
A Supply of 25x150 mm Test Tubes
Test Tube Racks for the above 25x150 mm Test Tubes
Gelman 47 mm Magnetic Vacuum Filter Funnel, 500 mL Vacuum Flask, and a Vacuum Supply
3. Reagents
Purity of Reagents—HPLC grade chemicals (where available) shall be used in all reagents for Ion
Chromatography, due to the vulnerability of the resin in the columns to organic and trace
metal contamination of active sites. The use of lesser purity chemicals will degrade the
columns.
Purity of Water—Unless otherwise indicated, references to water shall be understood to mean
Type I reagent grade water (Milli Q Water System) conforming to the requirements in
ASTM Specification D1193.
Eluent Preparation for SYSTEM2 TKN (TKN) Methods, including Total Nitrogen (using AG4A,
AG4A, and AS4A columns)—Weigh out 0.191 g of sodium carbonate (Na2CO3) and
0.143 g of sodium bicarbonate (NaHCO3) and dissolve in water. Bring the volume up to
1000 ml and place in the System 2 bottle labeled “IC-TKN 0.191/0.143”. Sparge the
eluent as in the above reagent for System 2.
100X Sample Spiking Eluent—prepared by using the above carbonate/bicarbonate ratios, but
increasing the concentration 100X. Weigh out 1.91 g of Na2CO3 and 1.43 g of NaHCO3
into a 100 mL volumetric flask. 0.05 mL of this solution is added to 5 mL of all samples
and standards to resolve the water dip associated with the fluoride peak.
Borate Buffer Solution---Dissolve 61.8 g H3BO3 and 8.0 g NaOH in a 1L volumetric flask
containing at least 500 mL of DI water. Swirl to mix and bring to volume. Make fresh
every 3 months.
Digestion Reagent---Dissolve 20.1 g of K2SO8 and 3.0 g of NaOH in a 1L flask containing at
least 500 mL of DI water. Swirl to mix and bring to volume. Make fresh every 3
months.
Quality Control---Commercially available wastewater TKN standard (Environmental Resource
Associates, “Ready-To-Use Wastewater QC Standards”, Cat # 743, Arvada, CO, 1800ERA-0122)
Glutamic Acid Stock Standard (C3H5NH2(COOH)2), 100 ppm---Dry Glutamic Acid in oven
at 1050C for 24 hours. Cover and place in dissector until cool. Dissolve 1.051g in
DI water and dilute to 1L; preserve with 2mL chloroform (CHCl3). Store in refrigerator
for no longer than 6 months.
Stock standard solutions, 1000 mg/L (1 mg/mL): Stock standard solutions may be purchased
(SPEX) as certified solutions or prepared from ACS reagent grade materials (dried at
105o C for 24 hours.
Nitrate Stock Standard (NO3-N), 1000 ppm--- dry Potassium Nitrate (KNO3) in oven ,
cover and place in dissector until cool. Dissolve 0.7218g in DI water and bring to 1 L;
preserve with 2 mL chloroform (CHCl3).
Nitrate Working Standard, 10 ppm---Dilute 100 mL of Nitrate Stock Standard to 1000
mL in 1L flask. Preserve with 2 mL chloroform (CHCl3). Store in refrigerator for no
longer than 6 months.
Calibration Standards—for the SYSTEM 2 TKN methods are prepared as
follows:
1. Using the 100 ppm Glutamic Acid Stock Standard, prepare the following:
a. 0.4 ppm = 1 mL of 100 ppm diluted to 250 mL
b. 0.8 ppm = 2 mL of 100 ppm diluted to 250 mL
c. 1.6 ppm = 4 mL of 100 ppm diluted to 250 mLl
2. Using the 10 ppm Nitrate Stock Standard, prepare the following:
a. 0.1 ppm = 1 mL of 10 ppm diluted to 100 mL
b. 0.2 ppm = 2 mL of 10 ppm diluted to 100 mL
c. 0.4 ppm = 4 mL of 10 ppm diluted to 100 mL
d. 0.8 ppm = 8 mL of 10 ppm diluted to 100 mL
e. 1.6 ppm = 16 mL of 10 ppm diluted to 100 mL
f. 2.9 ppm = 29 mL of 10 ppm diluted to 100 mL
3. The QC is diluted from the ordered solution: perform an appropriate dilution creating a
QC with a value on calibration curve, (~1.5 ppm) using the ordered standard.
4.
If it is deemed necessary, ICV’s (Initial Calibration Verification) and CCV’s (continuing
Calibration Verification) can be run using a 0.8 ppm and/or 1.6 ppm glutamic acid
solution.
Outside Source Certified Quality Control Sample—ERA
4. Procedure
A.
Instrument Preparation
1. Before turning on the Dionex Ion Chromatography System:
a. Fill the eluent reservoir(s) with fresh eluent.
b. Make certain the waste reservoir is empty of all waste.
c. Turn on the helium. The system pressure should be between 7 - 15psi. The system
pressure can be regulated with the knob on the back of the Eluent Organizer.
d. Connecting a piece of tubing to the gas line going into the eluent bottle and putting
the tubing into the eluent degasses the eluent reservoir(s). The gas knob on the
Eluent Organizer that corresponds to the eluent bottle should be slowly opened until
a constant bubbling stream can be seen in the eluent bottle.
e. The eluent should be degassed with helium, for a minimum of 30 minutes, before
operation of the instrument.
f. After the eluent has been degassed, remove the tube from the eluent and tightly seal
the eluent bottle. The eluent is now ready to introduce into the system.
2. Whether using the IP25 for Fluorides or the GP50 for everything else, turn off the
browser, scroll to REMOTE on the screen, select LOCAL and ENTER.
3. Scroll to mL/min., change to 0 mL/min., and hit ENTER. If using the IP25 pump, skip
to step #5.
4. Hit MENU and select 1, then ENTER.
5. Insert syringe into the Priming Block, open the gas valve on the Eluent Organizer, turn
the valve on the Priming Block counterclockwise, and turn on the pump that corresponds
with the method to be ran by pushing the OFF/ON button.
6. If the syringe does not fill freely, assist by gently pulling back on the plunger of the
syringe. Make certain that all of the air bubbles are removed from the eluent line to the
pumps.
7. Press OFF/ON on the pump to turn it off.
8. Turn the valve on the Priming Block clockwise, remove the syringe and expel the air
bubbles from the syringe.
9. Reinsert the syringe filled with eluent into the Priming Block.
10. Open the valve on the Pressure Transducer and the valve on the Priming Block with the
eluent filled syringe still attached. This is accomplished by turning both
counterclockwise.
11. Press PRIME on the pump and push the contents of the syringe into the Priming Block.
After the eluent has been injected into the Priming Block, press OFF/ON to turn the
prime pump off and to close the valves on the Pressure Transducer and Priming Block.
12. Remove the syringe from the Priming Block.
13. Scroll to the mL/min. on the screen for the pump. For the GP50, type 2 mL/min., and
press ENTER. For the IP25, type 1.2 mL/min., and press ENTER.
14. Press OFF/ON to turn on the pump at the appropriate rate. The pressure should soon
stabilize between both pumpheads after two minutes of pumping time.
15. If the pressure between pumpheads has a difference >20 psi, then shut down the pump
and repeat steps 2-14 to remove air bubbles and prime the pumps.
16. Once the pump has a pumping pressure difference between pumpheads of <20 psi, then
go to the computer and enter PeakNet.
17. On the computer, turn on the Chromeleon 6.8 browser, then choose System 2 (all
other anions including Bromide and TKN).
18. Go to last run sequence, click to highlight and go to file, click save as.. This will load
the method of interest and a template for the current sequence run.
19. The sequence is edited to reflect the method and samples that are to be run.
a. SYSTEM 2 TKN for TKN and Total Nitrogen
Note: Data is reprocessed in the section of Chromeleon 6.8 called Sequence integration
editor. Only operators with a minimum of three months experience in Ion
Chromatography should attempt to reprocess data for this analysis. Once data is
optimized, then the nitrogen values from nitrate and nitrite analysis can be subtracted
from this value for the TKN nitrogen value. If only Total Nitrogen is needed then use
the optimized data value without the correction for nitrite and nitrate nitrogen.
20. Observe the reading on the screen of the CD20 Conductivity Detector. A conductivity
rate change of <0.03 μS over a 30 second time span is considered stable for analysis.
21. If using the GP50 pump, it will take about 15-30 minutes for the CD20 system to
stabilize. If using the IP25, it will take between 30 minutes to 2 hours for stabilization.
22. Once the CD20 is stabilized, the Dionex DX500 Ion Chromatography
System is ready to start standardization.
NOTE: When using the GP50 Gradient Pump, all due care must be taken before one
switches from local procedures to remote procedures. The bottle from which the eluent
is being pumped (i.e., A, B, C, or D) must exactly match the bottle specified in the
method. If there is a difference, then once the pump control is turned over to remote
control, irreversible damage and destruction of suppressors, columns, piston seals, and
check valves on the GP50 Gradient Pump will occur. NEVER switch from bottle C to
A, B, or D without flushing the system lines with water to remove all traces of eluent
from bottle C from the lines.
B.
Sample Preparation
1. If the sample was not filtered in the field, it must be done so now. Transfer 50 mL of a
well-mixed sample to the filtering apparatus. Apply the suction and collect the filtrate.
2.
If the conductivity values for the sample are high, dilution will be necessary to properly
run the sample within the calibration standard range. Dilutions are made in the Polyvials
with the plastic Filter Caps. If the dilutions are > 20X, then volumetric glassware is
required.
3. All dilutions are performed with reagent grade DI water. Be sure to mix the dilution
well.
4. For the anions TKN and Total Nitrogen, first pipette 0.05 mL of 100X sample spiking
eluent into the Polyvials, then pipette 4.95 mL of the filtered samples on top of the
spiking eluent.
5. The Filter Caps are pressed into the Polyvials using the insertion tool.
6. Place the Polyvials into the Sample Cassette, which is placed into the Autosampler.
7. The white/black dot on the Sample Cassette should be located on right-hand side when
loaded in the left-hand side of the Automated Sampler for System 2.
8. For every ten samples the following should be included:
a. 1 DI water blank
b. 1 Duplicate of any one sample
c. 1 Quality Control sample/calibration check
C.
Calibration and Sample Analysis
1.
2.
3.
Set up the instrument with proper operating parameters established in the operation
condition procedure
The instrument must be allowed to become thermally stable before proceeding. This
usually takes 1 hour from the point on initial degassing to the stabilization of the baseline
conductivity.
To run samples on the Dionex Ion Chromatography System:
a. Make a run schedule on the PeakNet Software Section labeled SEQUENCE.
b. Double click the mouse on the SYS 2 to display the Scheduler Area.
The name of the calibration standards must be entered under the sample name
section
as Standard #1, Standard #2, and Standard #3.
Note: Level must be changed to the corresponding standard level or the calibration
will be in error. (Example: Standard #1 = Level #1; Standard #5 = Level #5)
c.
d.
4.
5.
6.
7.
Next, enter QC, blanks, QC, samples, duplicates, QC, and blanks, in that order.
Under sample type, click on either Calibration Standard or Sample, depending on
what is being run.
e. Under the Method section, the method name must be entered. To do so, double
click on the highlighted area under Method, scroll through the list of methods and
double click on the method of interest.
f. Next under the Data File section, enter the name of the data file.
g. Finally, in the Dil area, type in the dilution factor if different from 1. Do this for all
standards, blanks, quality controls, duplicates, and samples to be run under this
schedule.
h. Save the schedule and obtain a printout of it.
i. Standardize the Dionex Ion Chromatography System by running the standards:
Standard #1, Standard #2, and Standard #3.
Run the QC standards.
Run the prepblank and DI water blank.
Run the samples, duplicates, and blanks.
Run the QC standards at the end.
5. Calculations
A. Calculations are based upon the ratio of the peak area and concentration of standards to the
peak area for the unknown. Peaks at the same or approximately the same retention times are
compared. Once the method has been updated with the current calibration, this is calculated
automatically by the software using linear regression. Remember that when dilutions are
being run, the correct dilution factor must be entered.
B. Manual calculations are based upon the ratio of the peak and concentration of standards to the
peak area for the unknown when the software will not automatically calculate the unknown
concentration. Peaks at the same or approximately the same retention times are compared.
The unknown concentration can be calculated from using this ratio. Remember that when
dilutions are being run that the correct dilution factor must be entered before you will get the
correct result.
C. When possible the unknown should be bracketed between two knowns and the calculation of
the unknown made from both for comparison.
6. Quality Control
A quality control sample obtained from an outside source must first be used for the initial
verification of the calibration standards. A fresh portion of this sample should be analyzed
every week to monitor stability. If the results are not within +/- 10 % of the true value listed for
the control sample, prepare a new calibration standard and recalibrate the instrument. If this does
not correct the problem, prepare a new standard and repeat the calibration. A quality control
sample should be run at the beginning and end of each sample delivery group (SDG) or at the
frequency of one per every ten samples. The QC’s value should fall between ± 10 % of its
theoretical concentration.
A duplicate should be run for each SDG or at the frequency of one per every twenty samples,
whichever is greater. The RPD (Relative Percent Difference) should be less than 10%. If this
difference is exceeded, the duplicate must be reanalyzed.
From each pair of duplicate analytes (X1 and X2), calculate their RPD value:
⎛ X1 − X 2 ⎞
% RPD = 2 • ⎜
⎟ x 100
⎝ X1 + X 2 ⎠
where:
(X1 - X2) means the absolute difference between X1 and X2.
7. Method Performance
The method detection limit (MDL) should be established by determining seven replicates that are
2 to 5 times the instrument detection limit. The MDL is defined as the minimum concentration
that can be measured and reported with 99% confidence that the analyte concentration is greater
than zero and is determined from analysis of a sample in a given matrix containing the analyte.
MDL = t ( n −1,1−α = 99 ) ( S )
where:
t = the t statistic for n number of replicates used (for n=7, t=3.143)
n = number of replicates
S = standard deviation of replicates
8. Reference
EPA SW 846-9056, Chapter 5, September 1994
U.S. EPA Method 300.0, March 1984
ASTM vol. 11.01 (1996), D 4327, “Standard Test Method for Anions in Water by
Chemically
Suppressed Ion Chromatography”.
04/2011
KGS 4500-N C
Total Kjeldahl Nitrogen Preparation
1. Discussion
Principle
Total Kjeldahl Nitrogen is the sum of organic nitrogen and ammonia nitrogen compounds of a
sample. This method oxidizes all of the organic and inorganic nitrogenous compounds, at 100 to
110oC, to nitrate. The digestion also helps dissolve solid material that could interfere with
obtaining an accurate reading. The total nitrogen is then determined by the analysis of nitrate in
the digestate with an IC. Total Kjeldahl Nitrogen is then determined by subtracting the predetermined nitrite plus nitrate nitrogen values from the total nitrogen values.
Sensitivity
This method covers the range from 0.1 ppm to 2.9 ppm.
Interferences
Since this method is designed to oxidize ammonia to nitrate for analysis, the use of ammonia
and/or ammonia based substances should be avoided in the work area and on the glassware, as
this could produce increased positive results that are inaccurate.
Sample Preservation
This method cannot be performed on samples preserved in acid. Because of this, the samples
should be prepped ASAP.
2. Safety
Wear a lab coat, gloves, and protective eyewear when prepping this experiment to avoid possible
exposure to harmful substances.
3. Apparatus
CEM MARS Microwave Digestion Unit
Advanced Composite Vessels (ACV)
Graduated Cylinder
Wash Bottle
Automatic Pipettor
4. Reagents
Purity of Reagents—Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, all reagents shall conform to the specifications of the Committee on Analytical
Reagents of the American Chemical Society. Other grades may be used, provided it is
first ascertained that the reagent is sufficiently high in purity to permit its use without
lessening the accuracy of the determinations.
Purity of Water—Unless otherwise indicated, references to water shall be understood to mean
Type I reagent grade water (Milli Q Water System) conforming to the requirements in
ASTM Specification D1193.
Borate Buffer Solution—Dissolve 61.8 g H3BO3 and 8.0 g NaOH in a 1 L volumetric flask
containing at least 500 mL of DI water. Swirl to mix and bring to volume. Make fresh
every 3 months.
Digestion Reagent—Dissolve 20.1 g of K2S2O8 and 3 g of NaOH in a 1 L flask containing at least
500 mL of DI water. Swirl to mix and bring to volume. Make fresh every 3 months.
Quality Control—Commercially available wastewater TKN standard (Environmental Resource
Associates, “Ready-To-Use Wastewater QC Standards”, Cat # 743, Arvada CO, 1-800ERA-0122)
Glutamic Acid Stock Standard (C3H5NH2 (COOH) 2), 100 ppm—Dry Glutamic Acid in
oven at 105oC for 24 hours. Cover and place in dissector until cool. Dissolve 1.051g in
DI water and dilute to 1 L; preserve with 2 mL chloroform (CHCl3). Store in refrigerator
for no longer than 6 months.
Nitrate Stock Standard (NO3-N), 1000 ppm—Dry Potassium Nitrate (KNO3) in oven at 105oC for
24 hours. Cover and place in dissector until cool. Dissolve 0.7218g in DI water and
bring to 1 L; preserve with 2 mL chloroform (CHCl3). Store in refrigerator for no longer
than 6 months.
Nitrate Working Standard, 10 ppm—Dilute 100 mL of Nitrate Stock Standard to 1000 mL in 1 L
flask. Preserve with 2 mL chloroform (CHCl3). Store in refrigerator for no longer that 6
months.
5. Procedure
A. Turn on the CEM MSP 1000 Microwave Digestion Unit and allow it to warm up for at least
15 minutes.
B.
Standards Prep
1. Using the 100 ppm Glutamic Acid Stock Standard, prepare the following:
a. 0.4 ppm = 1 mL of 100 ppm diluted to 250 mL
b. 0.8 ppm = 2 mL of 100 ppm diluted to 250 mL
c. 1.6 ppm = 4 mL of 100 ppm diluted to 250 mL
2. Using the 10 ppm Nitric Stock Standard, prepare the following:
a. 0.1 ppm = 1 mL of 10 ppm diluted to 100 mL
b. 0.2 ppm = 2 mL of 10 ppm diluted to 100 mL
c. 0.4 ppm = 4 mL of 10 ppm diluted to 100 mL
d. 0.8 ppm = 8 mL of 10 ppm diluted to 100 mL
e. 1.6 ppm = 16 mL of 10 ppm diluted to 100 mL
f. 2.9 ppm = 29 mL of 10 ppm diluted to 100 mL
3. The QC is diluted from the ordered solution: perform an appropriate dilution creating a
QC with a value midway on calibration curve, (~1.5 ppm) using the ordered standard.
4. If it is deemed necessary, ICV’s (Initial Calibration Verification) and CCV’s (Continued
Calibration Verification) can be run using a 0.8 ppm and/or 1.6 ppm glutamic acid
solution.
C.
Sample Prep
1. The Prep Blank is 10 mL of reagent grade DI water poured into the first liner.
2. For all samples and QC, a 10 mL aliquot should be poured into one of the advanced
composite vessels, or ACV, liners.
3. Add 5 mL of Digestion Reagent to each liner.
4. Assemble the ACV system as described in Microwave Digestion GLP.
D.
Digestion Set Up
1. From the options on the main menu of the microwave, press F3—“Recall Method/Data”.
2. Press F1—“Recall Stored Method”.
3. Use arrow keys to scroll down to “TKN SM”; press “Enter”.
4. Press F1—“Load Program”.
5. Press F4—“Start”.
6. Press F1—“Yes”. Once a digestion is started, watch the temperature probe and pressure
tube carefully to make sure they do not become tangled up. If they do become tangled,
press F1 to abort the run and remedy the problem.
7. Once the run is complete, disassemble the ACV’s, add 1 mL of Borate Buffer Solution to
each liner ( all QC, samples, dups., etc.) and pour the digested samples into appropriately
labeled precleaned containers.
8. The digested QC and samples, along with the corresponding data sheets, are to be
transferred to the IC for analysis.
E.
Prep-Batching
1. Log-on to the “Labworks” system.
2. Click on “Edit Data”.
3. Enter the SDG number or choose it from the list.
4. Click on “OK”.
5. Click on “OK”.
6. In the row for TKN prep work (TKN_PREP), enter a 1 under the number of each sample
completed and save it.
7. Exit system.
6. Quality Control
A duplicate sample should be prepped at the frequency of one per every twenty samples
(sufficient sample permitting), or one per SDG, whichever is greater. The RPD should be less than
10%. If this difference is exceeded, the duplicate may need to be reprepped. The QC’s value
should fall between ± 10 % of its theoretical concentration as well.
7. References
Standard Methods for the Examination of Water and Wastewater, 20th edition (1998),
Method 4500-N C, pg. 4-102
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