Chemical Hygiene Plan - QCNR
Chemical Hygiene Plan
Important Telephone Numbers:
1. 911 for All Emergencies
2. (435) 797-1939 Utah State University Police Department (Non-Emergency Line)
3. (435) 716-9500 Logan Fire Department (Non-Emergency Line)
1. Purpose
This chemical hygiene plan establishes a formal written program for protecting users against
health and safety hazards associated with exposure to hazardous chemicals or field conditions
that may lead to accident or injury. This chemical hygiene plan (CHP) describes the proper use
and handling procedures to be followed by faculty, staff, and all other personnel working with
hazardous chemicals in laboratory settings (NR 309) or in the field (Green Canyon Ecology
Center and Irrigated Pasture Project).
2. Employee Rights and Responsibilities
Employees and other personnel who work at the aforementioned locations have the right to be
informed about the potential hazards of the chemicals in their work areas and to be properly
trained to work safely with these substances. All personnel, including principal investigators,
laboratory supervisors, laboratory technicians, student workers, and support staff have a
responsibility to maintain a safe work environment. All personnel working with chemicals are
responsible for staying informed on the chemicals in their work areas, safe work practices and
SOPs, and proper personal protective equipment (PPE) required for the safe performance of
their laboratory work.
A. Laboratory Supervisor Responsibilities
The Laboratory Supervisor is the individual that is ultimately responsible for the overall
laboratory and field operations, ensuring that the requirements of the CHP are followed by all
staff members that work in the lab and in the field. The Principal Investigator (PI), Juan Villalba,
is the Laboratory and Field Supervisor. The Laboratory and Field Supervisor may delegate some
safety duties to a qualified individual, but ultimately remains responsible for the safety of all
personnel working in the laboratory. Specifically, the Supervisor must:
1. Understand applicable environmental health and safety rules, including the contents of the
CHP;
2. Identify hazardous conditions or operations in the laboratory and field and establish SOPs
and hazard assessments to effectively control or reduce hazards;
3. Ensure that all laboratory personnel that work with hazardous chemicals receive appropriate
training (see https://rgs.usu.edu/ehs/laboratory-safety-initial/ for laboratory safety training at
USU and lab training registration)
4. Maintain written records of lab-specific training
5. Ensure that appropriate Personal Protective Equipment (PPE) (e.g., laboratory coats, gloves,
eye protection, etc.,) and engineering control equipment (e.g., chemical fume hood) are made
available, in good working order, and being used properly.
6. Conduct periodic lab inspections and immediately take steps to abate hazards that may pose
a risk to life or safety upon discovery of such hazards; and
B. Laboratory Employee Responsibilities
All employees (e.g., graduate research assistants, graduate students, undergraduate students,
lab technicians, post-doctoral researchers, and visiting scientists) in laboratories that use,
handle, or store hazardous chemicals must:
1. Review and follow the requirements of the CHP.
2. Follow all verbal and written laboratory safety rules, regulations, and SOPs required for the
tasks assigned.
3. Develop and practice good personal chemical hygiene habits such keeping work areas clean
and uncluttered.
4. Plan, review, and understand the hazards of materials and processes in the laboratory prior
to conducting work.
5. Utilize appropriate measures to control hazards, including consistent and proper use of
engineering controls, administrative controls, and PPE.
6. Understand the capabilities and limitations of PPE.
7. Immediately report all accidents, near misses, and unsafe conditions to the Laboratory
Supervisor.
8. Complete all required mandatory safety training and provide written documentation to the
Laboratory Supervisor;
9. Inform the Laboratory Supervisor of any work modifications ordered by a physician as a
result of medical surveillance, occupational injury, or chemical exposure.
3. Hazardous Chemical Classification Systems
Chemical classification systems are designed to communicate hazards. The three most widely
used classification systems are the OSHA Globally Harmonized System for Classifying and
Labeling Chemicals (implemented under the OSHA Hazard Communication Standard), the
National Fire Protection Association (NFPA) system of classifying the severity of hazards, and
the Department of Transportation (DOT) hazard classes. These classification systems are used
by chemical manufacturers when creating safety data sheets and chemical labels, therefore it is
important that lab employees understand the basic elements of each classification system.
A. Globally Harmonized System for Classifying Chemicals
The Globally Harmonized System (GHS) is a world-wide system adopted by OSHA for
standardizing and harmonizing the classification and labeling of chemicals. The objectives of the
GHS are to:
-Define health, physical, and environmental hazards of chemicals;
-Create classification processes that use available data on chemicals for comparison with the
defined hazard criteria (numerical hazard classification is based on a 1 – 5 scale, 1 being the
most hazardous and 5 being the least hazardous); and
-Communicate hazard information, as well as protective measures, on labels and Safety Data
Sheet (SDS), formerly known as Material Safety Data Sheets (MSDS).
Safety Data Sheets
The SDS provides comprehensive information that is imperative for the safe handling of
hazardous chemicals. Laboratory personnel should use the SDS as a resource to obtain
information about hazards and safety precautions. SDSs cannot provide information for hazards
in all circumstances. However, the SDS information enables the employer to develop an active
program of worker protection measures such as training on hazard mitigation. Chemical
manufacturers are required to use a standard format when developing SDSs. The SDS will
contain 16 headings which are illustrated in Figure 1: GHS Required sections of a safety data
sheet.
Chemical Labeling
The GHS standardized label elements, which are not
subject to variation and must appear on the chemical
label, contain the following elements:
1) Symbols (hazard pictograms) are used to convey health,
physical and environmental hazard information, assigned to a GHS hazard class and category;
2) Signal Words such as “Danger" (for more severe hazards) or "Warning" (for less severe
hazards), are used to emphasize hazards and indicate the relative level of severity of the hazard
assigned to a GHS hazard class and category;
3) Hazard statements (e.g., “Danger! Extremely Flammable Liquid and Vapor”) are standard
phrases assigned to a hazard class and category that describe the nature of the hazard; and
4) Precautionary statements are recommended measures that should be taken to minimize or
prevent adverse effects resulting from exposure to the hazardous chemical.
Figure 2. GHS Hazard Pictograms
Figure 3. GHS Hazard classification system for flammable
liquids
B. National Fire Protection Association Rating System
The NFPA system uses a diamond-shaped diagram of symbols and numbers to indicate the
degree of hazard associated with a particular chemical. This system was created to easily and
quickly communicate hazards to first responders in the event of an emergency situation. These
diamond-shaped symbols are placed on chemical containers to identify the degree of hazard
associated with the specific chemical or chemical mixture. The NFPA system is a common way
to identify chemical hazards and should be understood by laboratory employees. The NFPA 704
numerical rating system is based on a 0 – 4 system; 0 meaning no hazard and 4 meaning the
most hazardous (note: this in contrast to the GHS system of 1 –5 where 1 is the most hazardous
and 5 is the least hazardous). Figure 2.4 illustrates the NFPA hazard rating system and identifies
both the hazard categories and hazard rating system.
Figure 4. NFPA Hazard Rating System
C. Department of Transportation (DOT) Hazard
Classes
The DOT regulates the transportation of all hazardous
materials in the United States, and defines a
hazardous material as any substance that has been
determined to be capable of posing an unreasonable
risk to health, safety, or property when transported
in commerce.
Chemical Stores (435-797-1616;
http://www.chem.usu.edu/facilities/chem_stores
will help with transportation of hazardous materials
on campus.
4. RECOMMENDED CHEMICAL SPILL CLEANUP PROCEDURES
YOU SHOULD NOT CLEAN UP A SPILL IF:
•
You don’t know what the spilled material is
•
You lack the necessary protection or equipment to do the job safely
•
The spill is too large to contain
•
The spilled material is highly toxic
•
You feel any symptoms of exposure
Instead contact: (https://rgs.usu.edu/ehs/ehs-tools/) Under accident reporting. OR, if
immediately health-threatening call 911 (campus phone)
SPILL RESPONSE SCHEME
Evaluate and Notify
•
Assess the toxicity, flammability, or other properties of material (see label & MSDS)
•
For flammables, remove or turn off ignition sources such as motors, pumps, fridges.
•
Determine if there is an immediate health threat to you or your neighbors. If so, alert
neighbors, isolate the area and call for help using the phone numbers above.
•
If spill is minor, begin cleanup following steps below
Containment/Cleanup
•
Don appropriate gloves, eye protection, lab coat, etc.
•
Per SDS use absorbents* (e.g., “spill pillows” for solvents), or neutralizers appropriate for
the material*, e.g. sodium bicarbonate for acids, citric for bases.
•
Protect floor drains with absorbents or barriers around them
•
Package and label waste. Include contaminated clothes, rags, equipment, etc.
•
Store temporarily in a fume hood if material is volatile
Followup
•
•
•
Send a Haz. Pickup request (https://rgs.usu.edu/ehs/ehs-tools/)
Reorder and restock cleanup materials used
Inform EHS if there were any personnel exposures, or release to the environment
SELECTING THE PROPER GLOVES
All gloves are permeable, only the permeation rate varies, depending on the chemical, the
glove material and thickness, temperature, concentration gradient, etc. However, once a
material begins to permeate the glove, it will continue until an equilibrium is reached. You
must, therefore, decide when it is appropriate to discard dirty gloves.
Check gloves before use for signs of wear or penetration. Disposable gloves can be inflated to
check for pinholes. When removing gloves, be careful to avoid touching the outside of the
gloves with your bare hands. Always remove gloves before leaving lab.
Disposable gloves provide minimal protection and should be used accordingly. If using
concentrated solvents, corrosives or toxics, more heavy-duty gloves should be worn. These
provide more protection, but have the drawback of being more cumbersome. Note also that
about 15% of the population is allergic to latex to some degree.
This website suggests an appropriate glove as a function of the chemical in use:
https://www.coleparmer.com/safety-glove-chemicalcompatibility?PubID=WU&persist=True&gclid=CKG0xKyJp9ECFQ90fgod1YEPGA
Safety Data Sheets (Formerly known as MSDS)
http://www.sigmaaldrich.com/safety-center.html
https://www.fishersci.com/us/en/catalog/search/sdshome.html
CHEMICAL HIGIENE PLAN -Specific Plan
Laboratory Location: NR 309
Supervisor Name: Juan Villalba
Laboratory PI or Supervisor Signoff (required). I certify that I have reviewed and approve
the attached Laboratory-specific Chemical Hygiene Plan with Standard Operating Procedures
for my above laboratory locations.
Signature:________________________________________
Laboratory Worker Training Record: Chemical Hygiene Plan
Principal Investigator/Supervisor: _________________________________
The following lab workers have reviewed and understand the following elements of the
attached Chemical Hygiene Plan for the above PI or supervisor:
•
Sec. I: Laboratory-specific Chemical Hygiene Plan with Standard Operating Procedures
•
Sec. II: Online Safety Training (https://rgs.usu.edu/ehs/laboratory/)
Name (print)
Signature
Date
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Standard Operating Procedures – Lab 309
Standard Operating Procedure – Use of Freeze Dryer
Section 1 - Personal Protective Equipment
1. Lab Coat or Lab Gown
2. Hair tied back if long
3. Proper enclosed footwear
4. Safety glasses or goggles
5. Nitrile or latex gloves
Section 2 – Potential Hazards + Safety precautions
1. Freeze dryer generates a high vacuum which can cause implosion of glass ampoules – this
poses a threat especially to eyes – wear safety glasses at all times when using the freeze
dryer.
2. Glass ampoules can break if mishandled, and the sharps generated pose a risk of cuts – such
cuts are especially dangerous if hazardous microbes are present in the ampoules. Handle
ampoules with care at all times, and know the risks and properties of the microbes you are
working on.
3. Freeze dryer generates low temperatures. Ampoules and other parts of the machine may
cause cold burns if touched to exposed skin. Wear gloves when handling these cold parts, or
avoid contact.
4. Freeze dryer is an electrical device and thus poses electrical shock hazard if misused or if
faulty. Inspect machine for any problems before use (e.g. problems with power supply, sparks,
burning smell etc). If any problems present, do not use machine, notify service centre and/or
manufacturer. Do not allow water to enter the internal parts of the machine. Keep flammables
away from the machine.
5. Do not use freeze drier for evaporating samples containing organic solvents!
6. Do not use the freeze dryer unless an experienced user has shown you in detail how to use it.
Standard Operating Procedure – Digestibility Using the Gas Production Technique
Section 1 - Personal Protective Equipment
1. Lab Coat or Lab Gown
2. Hair tied back if long
3. Proper enclosed footwear
4. Safety glasses or goggles
5. Nitrile or latex gloves
6. Palpation gloves
Section 2 – Potential Hazards + Safety precautions
Rumen fluid extraction: Restrain animal in a cattle chute. Stay always behind a fence before
the animal is restrained. Open the canula, use palpation gloves and introduce a ½ L cup in the
ventral area of the rumen environment to collect rumen fluid. Fill a 2 L thermos with rumen
fluid and transport immediately to the lab.
Incubation:
1. Set temperature for the incubator at 38°C.
2. Check the reactor bottles for cracks and other damages. Be careful to prevent cuts if bottles
are broken. Discard broken bottles.
3. Fill inoculum, substrate/cellulose and buffer into the glass reactors according to the
experimental set-up. Shake the bottle gently to prevent clumping.
4. Control the pH-value, recommended values are between 7.5 and 8.
5. Close the bottle using the rubber plug (with attached cannula) and the screw cap. Do it gently
to prevent breaking the glass.
6. Glass reactors can break if mishandled, and the sharps generated pose a risk of cuts. Handle
reactors with care at all times.
7. The reactor flasks are put under vacuum app. 0.7 bar. Close the cock and control the stability
of the vacuum (pressure tightness) using the manometer.
8. The reactor will receive a small over pressure of CO2, which will be discharged just
connecting a needle to the manometer. Be careful when handling the needle.
9. Measure the pressure on the reactors every day or as often as possible (wear safety goggles!)
by connecting the manometer on top of the reactor and introducing the needle. Set the
manometer to zero before (atmospheric). Also make sure to conduct a fast measurement,
so the gas inside the reactor cannot cool down to lose volume.
10. Pressures inside the reactors should not exceed 1 bar over atmospheric due to security
reasons. If the measured pressure approaches this value the gas has to be discharged.
11. Discharge the gas left inside after the experiment period, remove the screw lid and the
rubber plug and rinse everything with water.
Standard Operating Procedure – weigh of Cr2O3 for intake estimates
Section 1 - Personal Protective Equipment
1. Lab Coat or Lab Gown
2. Hair tied back if long
3. Proper enclosed footwear
4. Safety glasses or goggles
5. Nitrile or latex gloves
Section 2 – Potential Hazards + Safety precautions
MSDS-Cr2O3
Hazards Identification
Potential Acute Health Effects: Hazardous in case of skin contact (irritant), of eye contact
(irritant), of ingestion, of inhalation.
Potential Chronic Health Effects: Hazardous in case of inhalation. Slightly hazardous in case of
skin contact (sensitizer). CARCINOGENIC EFFECTS: A4
(Not classifiable for human) by ACGIH, 3 (Not classifiable for human.) by IARC. MUTAGENIC
EFFECTS: Mutagenic for mammalian somatic cells. Mutagenic for bacteria and/or yeast.
TERATOGENIC EFFECTS: Not available.
DEVELOPMENTAL TOXICITY: Not available. The substance may be toxic to upper respiratory
tract, skin. Repeated or prolonged exposure to the substance can produce target organs
damage.
First Aid Measures
General advice
Consult a physician. Show this safety data sheet to the doctor in attendance.
If inhaled
If breathed in, move person into fresh air. If not breathing, give artificial respiration. Consult a
physician.
In case of skin contact
Wash off with soap and plenty of water. Consult a physician.
Eye Contact:
Check for and remove any contact lenses. In case of contact, immediately flush eyes with plenty
of water for at least 15 minutes. Get medical attention.
Skin Contact:
In case of contact, immediately flush skin with plenty of water. Cover the irritated skin with an
emollient. Remove contaminated clothing and shoes. Wash clothing before reuse. Thoroughly
clean shoes before reuse. Get medical attention.
Serious Skin Contact:
Wash with a disinfectant soap and cover the contaminated skin with an anti-bacterial cream.
Seek medical attention.
Inhalation:
If inhaled, remove to fresh air. If not breathing, give artificial respiration. If breathing is difficult,
give oxygen. Get medical attention.
Serious Inhalation: Not available.
Ingestion:
Do NOT induce vomiting unless directed to do so by medical personnel. Never give anything by
mouth to an unconscious person. If large quantities of this material are swallowed, call a
physician immediately. Loosen tight clothing such as a collar, tie, belt or waistband.
Serious Ingestion: Not available.
Most important symptoms and effects, both acute and delayed
The most important known symptoms and effects are described in the labelling (see section
2.2) and/or in section 11
Indication of any immediate medical attention and special treatment needed
No data available
Firefighting Measures
Suitable extinguishing media
Use water spray, alcohol-resistant foam, dry chemical or carbon dioxide.
5.2 Special hazards arising from the substance or mixture
No data available
Advice for firefighters
Wear self-contained breathing apparatus for firefighting if necessary.
Accidental Release Measures
Personal precautions, protective equipment and emergency procedures
Use personal protective equipment. Avoid dust formation. Avoid breathing vapours, mist or
gas. Ensure adequate ventilation. Avoid breathing dust.
6.2 Environmental precautions
Do not let product enter drains.
Methods and materials for containment and cleaning up
Pick up and arrange disposal without creating dust. Sweep up and shovel. Keep in suitable,
closed containers for disposal.
Handling and Storage
Further processing of solid materials may result in the formation of combustible dusts. The
potential for combustible dust formation should be taken into consideration before additional
processing occurs. Provide appropriate exhaust ventilation at places where dust is formed.
For precautions see section 2.2.
Conditions for safe storage, including any incompatibilities
Keep container tightly closed in a dry and well-ventilated place
Hygroscopic Handle and store under inert gas. Keep in a dry place.
Storage class (TRGS 510): Non Combustible Solids
Exposure controls
Appropriate engineering controls
Handle in accordance with good industrial hygiene and safety practice. Wash hands before
breaks and at the end of workday.
Personal protective equipment
Eye/face protection
Safety glasses with side-shields conforming to EN166 Use equipment for eye protection tested
and approved under appropriate government standards such as NIOSH (US) or EN 166(EU)
Skin protection
Handle with gloves. Gloves must be inspected prior to use. Use proper glove removal technique
(without touching glove's outer surface) to avoid skin contact with this product. Dispose of
contaminated gloves after use in accordance with applicable laws and good laboratory
practices. Wash and dry hands.
Material: Nitrile rubber
Minimum layer thickness: 0.11 mm
Break through time: 480 min
Material tested:Dermatril® (KCL 740 / Aldrich Z677272, Size M)
Splash contact
Material: Nitrile rubber
Minimum layer thickness: 0.11 mm
Break through time: 480 min
Material tested:Dermatril® (KCL 740 / Aldrich Z677272, Size M)
data source: KCL GmbH, D-36124 Eichenzell, phone +49 (0)6659 87300, e-mail [email protected],
test method:
EN374
If used in solution, or mixed with other substances, and under conditions which differ from EN
374, contact the supplier of the CE approved gloves. This recommendation is advisory only and
must be evaluated by an industrial hygienist and safety officer familiar with the specific
situation of anticipated use by our customers. It should not be construed as offering an
approval for any specific use scenario.
Body Protection
Choose body protection in relation to its type, to the concentration and amount of dangerous
substances, and to the specific work-place., The type of protective equipment must be selected
according to the concentration and amount of the dangerous substance at the specific
workplace.
Respiratory protection
Respiratory protection is not required. Where protection from nuisance levels of dusts are
desired, use type N95 (US) or type P1 (EN 143) dust masks. Use respirators and components
tested and approved under appropriate government standards such as NIOSH (US) or CEN (EU).
Control of environmental exposure
Do not let product enter drains.
Standard Operating Procedure – Condensed Tannin Analyses
Section 1 - Personal Protective Equipment
1. Lab Coat or Lab Gown
2. Hair tied back if long
3. Proper enclosed footwear
4. Safety glasses or goggles
5. Nitrile or latex gloves
Determination of Total Condensed Tannins
This procedure is an adaptation of method for determining total condensed tannins developed
by Terrill et al. (1992). The method quantifies both tannins that can be extracted by organic
solvents such as acetone (extractable condensed tannins) as well as those that remain bound to
the plant tissue after treatment with organic solvents (bound condensed tannins). This
procedure uses 70% acetone as a solvent which has been shown to give more complete and
repeatable tannin extraction than other solvents (Terrill et al. 1990).
This procedure is a colorimetric method based on butanol-HCl procedure. The primary classes
of condensed tannins found in forage legumes are Procyanidins and Prodelphinidins which are
composed of chains of cyanidin and delphinidin subunits respectively. When placed in a heated
butanol-HCL solution the condensed tannins break down into cyanidin and delphinidin subunits
(Porter et al. 1986). These cyanidin and delphinidin subunits show up as red, pink, or purple
colorants in the butanol-HCL solution, with the exact color varying depending upon the relative
percentage of cyanidin and delphinidin subunits present. Therefore the concentration of these
cyanidin and prodelphinidin subunits can be measured by a spectrophotometer.
Purified tannin from the plant species being analyzed is used as the standard for determining
tannin content. There are two reasons for doing so. First, using purified tannin is considered to
give results that are more accurate and “biologically relevant” than those obtained using
catechin (Terrill et al. 1990). Second, using commercial preparations such as tannic acid, gallic
acid or catechin introduces repeatability problems associated with product variation between
manufacturers. Makkar and Becker (1994) found variations between different suppliers of
tannic acid could result in a 20%-30% difference in calculated tannin content of a given plant
sample, thus making it difficult to make meaningful comparisons between results obtained by
different labs using tannic acid from different suppliers.
I. Sample Preparation
When samples are collected in the field they should be placed immediately on dry ice and
transported to a deep freeze or directly to a freeze drier. Samples should not be allowed to
thaw between the time they are collected and freeze-dried. Allowing samples to thaw reduces
tannin extractability as does air-drying or oven-drying (Terrill et al. 1990). After samples are
freeze-dried they should be ground in Wiley mill to pass through a 1mm screen and stored in a
cool, dry place.
II. Isolating the Purified Tannin for Standards
To obtain the purified tannin it is necessary to “sacrifice” 100-300 grams of ground fresh or
freeze-dried plant material. For high tannin species like sericea lespedeza 100 grams will be
sufficient, but 300 grams may be needed for low-tannin species such as birdsfoot trefoil. Final
yield of pure tannin will be about 20% of the extractable tannin content of the forage. For
example 100 grams of sericea lespedeza with 10% extractable tannin content will yield about 2
grams of purified tannin. The procedure needs advanced planning. The sephadex LH-20 beads
need to be soaked overnight prior to the procedure. The procedure not counting freeze-drying
will take 1-2 people approximately 16 hours. The procedure will require 100 grams of sephadex
LH-20 beads, 1-2 liters of diethyl ether, 2-3 liters of methanol and 4-6 liters of acetone.
A.) Extraction of tannin from plant material.
1) Mix the ground plant material in a beaker with a solution of 70% acetone, 30% water. Stir
thoroughly and pour mixture into 50 ml or larger centrifuge tubes and centrifuge until the
solids settle out. Pour off the supernatant (liquid) and save.
2) Pour additional 70% acetone over the pellet (solid) from the above step, vortex and recentrifuge. Save the supernatant and combine with that from the first extraction. Two
extractions will extract the majority of tannins but a third extraction will slightly increase the
total tannin yield.
B.) Removal of Plant Pigments
1) Pour the acetone extract obtained above into a large separation funnel. Add 1 ml of diethyl
ether for every 2 ml of acetone extract. Mix until a phase separation occurs. Draw off and save
the lower aqueous layer (light brown or yellow) which contains the tannin and discard the
upper organic layer containing plant oils, chlorophyll and other pigments.
2) Repeat 3-4 times to remove all plant pigments and oils.
C.) Removal of Organic Solvents
Take aqueous layer obtained above and place in a rotary evaporator. Start out at a relatively
low temperature until all of the diethyl ether is removed then increase heat to remove acetone
residues. If any solids remain in liquid they should here be removed by centrifugation.
Measure and record the volume of aqueous solution.
D.) Removal of Non-tannin Phenolics
In this step you will “adhere” the tannin and non-tannin phenolics to the sephadex beads and
then remove the non-tannin phenolics with 50% methanol.
1) Pour 100 gram of sephadex LH-20 beads into a beaker and add 550 ml distilled water and
soak overnight. The next day pour off the excess water into a graduated cylinder and record
volume. The difference between the volume added and the volume removed will be the
volume absorbed by the beads.
2) Combine swollen sephadex beads with the aqueous solution from section C in a large beaker
and add an amount of methanol equal to the amount of aqueous solution plus the amount of
water absorbed by the sephadex beads. Stir this solution for 30 minutes and then let set for 2-3
hours to allow the tannin to adhere to the beads.
3) Mix up 2 liters of 50% methanol 50% distilled water solution.
4) Set up a large Buchner funnel on a vacuum filtration apparatus, cover bottom with filter
paper and moisten with 50% methanol and begin vacuum. Pour the sephadex/water/methanol
solution from step #2 into the Buchner funnel. Wash the beads until the liquid coming off them
is completely clear. Try to avoid letting the beads get sucked dry during this process. This
process will take 1-3 hours and will require 1-2 liters of 50% methanol. Discard the liquid.
E.) Recovery of Tannin from Beads
1) After discarding the methanol rinse above, rinse the vacuum bottle well with acetone and
place the Buchner funnel with beads back on the vacuum bottle.
2) Rinse the sephadex beads with 70% acetone, 30% water until the beads are snowy white.
This should take about 1-1.5 liters of the 70% acetone. SAVE the liquid.
F.) Removal of Acetone and Water and Recovery of Pure Tannin
1) Take liquid rinse above and place in rotary evaporator to remove all traces acetone and
methanol.
2) After organic solvents have been removed, pour the liquid out in a shallow layer in metal
freeze-drier pans. Place pans in deep freeze overnight. The next day place pans into freeze
drier. It should take about a week for removal of all the water leaving the purified tannin which
looks like cotton candy and will be anywhere from off-white to reddish-brown depending upon
the plant species. Store tannin in a cold, dry place until it is needed.
III. Determining Extractable Condensed Tannins
A.) Preparation of 70% acetone solution.
1) Measure out 740 ml of HPLC grade acetone and 310 ml of distilled water. This will yield one
liter of mix due to 5% loss in volume that occurs when acetone and water are mixed.
2) Weigh out 1 gram of ascorbic acid and dissolve in the liter of acetone/water mix.
B.) Preparation of the butanol-HCl solution.
Measure out 950 ml of 1-butanol, and add 50 ml of concentrated (37%) HCl. Due the hazards
involved with handling concentrated acids this step should be performed under a fume hood
and appropriate eye and skin protection equipment should be worn. Always add acid to
butanol, not butanol to acid.
C.) Weighing sub-samples
Weigh and record triplicate 500 mg sub-samples of each plant sample
D.) Extracting samples
1) Pipette 20 ml of the 70% acetone mix into each centrifuge tube, cap and vortex mix each
tube twice for 10 seconds each time.
2) Place into heated shaking bath (32 degrees Celsius, 95 RPM) for 15 minutes.
3) Remove tubes from bath, briefly hand mix and centrifuge for 15 minutes at 900 G’s. This
corresponds to the 40 setting on the international model K centrifuge in our lab. RCF in G’s is
calculated by the formula G = (11.17 x 10-7) RN2 where R is radius in mm from center of
centrifuge spindle the far tip of centrifuge tube and N is the speed of the spindle in RPM.
4) Decant liquid into labeled beakers and cover with parafilm.
5) Repeat steps 1-4 twice for a total of three extractions.
6) Combine the three extracts and pour into a 100 ml graduated cylinder. Use an eyedropper to
add 70% acetone solution to bring volume to nearest whole ml. Record volume in notebook
and pour into new 50 ml screwtop centrifuge tubes. (Final volume will be 55-60 ml so a small
amount of extract will be discarded).
7) Place extracts in freezer until you are ready to perform the butanol-HCl or Folin-Ciocalteu
analyses. Put the residues in the freezer as well to save for the bound condensed tannin
analysis. Extracts will keep up to 30 days in freezer.
E.) Determining Extractable Condensed tannins using the butanol-HCl procedure
1) To prepare the 1mg/ml stock solution used in the standard curve, weigh out 100 mg of pure
tannin, dissolve in 50ml of 70% acetone and carefully pour into a 100 ml volumetric flask. Then
bring volume up to exactly 100 ml. Allow to “equilibrate” in refrigerator for 3 hours before
using. The stock solution will keep for up to a month as long as it is kept refrigerated and
capped.
2) To construct the standard curve pipette 20 ul (microliters) of the stock solution into each of
16 ml screwcap test tubes and do the same likewise for the 50, 100, 200, and 250 ul
concentrations.
3) Pipette 100 ul of each extraction obtained in section D into triplicate 16 ml screwtop test
tubes. This will give a 3 X 3 subsampling regime--three subsamples taken from each sample and
3 absorbance values for each subsample.
4) Pipette 5 ml of butanol-HCl into each tube containing the stock solution and plant sample
extracts and mix with a vortex mixer. Also prepare a blank tube with 5 ml of butanol-HCl
solution.
5) Place in a 95 degree Celsius water bath and cap each tube with a marble.
6) After 1 hour and 15 minutes remove tubes from bath and cool them in ice water.
7) Set the spectrophotometer at 550 nm, with filter appropriate for 550nm wavelength and
“blank” the machine with the blank tube.
8) Record the absorbance of each of the standard curve and sample tubes.
9) Calculate the percent extractable condensed tannins by calculating mg/ml based on the
standard curve equation, multiplying by ten (100 ul is 0.1 ml) then multiplying by ml of extract
to total mg grams of extractable tannin. Then divide mg extractable tannin by sample weight
and multiply by 100 to get percent extractable tannin. Once the dry matter of the freeze-dried
sample is determined, the numbers can be corrected to a 100% dry matter basis.
IV. Determining Bound Condensed Tannins
A.) Recovery of residues from the acetone extraction.
1.) Weigh a 150 mm Whatman #1 filter paper to establish a tare weight and write sample
number on it with a lead pencil. Due to variation in filter weights it is necessary to pre-weigh
each filter.
2) Fold and place inside a funnel on a vacuum apparatus. Wet the filter down with 100%
acetone. Dump the residue from procedure III.-D above, onto the filter and thoroughly rinse
out residue centrifuge tube onto the filter with 100% acetone.
3) Thoroughly wash the residue in the filter with 100% acetone. Maintain vacuum until sample
and filter are both dry.
4) Allow filter and sample to air dry for an additional two hours.
5) Weigh the dried filter with residue and calculate the net weight of residue.
6) Place residues back in the centrifuge tubes they came from, making sure that the tubes are
thoroughly dried.
B.) Measuring bound condensed tannins with the butanol-HCl procedure.
1.) Weigh triplicate 10 mg samples of residue and place in 16 ml screw-top test tubes. This
again is a 3 X 3 subsampling regime, three subsamples taken from each sample and 3
absorbance values recorded for each subsample.
2) Construct a standard curve with the 1 mg/ml stock solution and a blank butanol-HCl tube as
in section III.-E.
3) Pipette 5 ml of butanol-HCl into each residue tube and mix with vortex mixer.
4) Place all tubes into 95 degree Celsius water bath for one hour and 15 minutes.
5) Cool tubes in ice water.
6) Remix the residue tubes with vortex mixer. For high-tannin samples such as sericea
lespedeza it may be necessary to add another 5 ml of butanol-HCl to keep absorbance values
within the range that can be read by the spectrophotometer. Make appropriate adjustments in
calculations.
7) Centrifuge the residue tubes for 15 minutes at 350 G’s using special inserts to hold the tubes
in place.
8) Record absorbance values for each tube on spectrophotometer at 550 nm.
9) Calculate bound condensed tannins by calculating grams of tannin via standard curve
equation then dividing by proportion of residue sampled to get mg of bound condensed tannin
in samples, and dividing by sample weight to get percent bound condensed tannins. When dry
matter of samples is determined, convert to a dry matter basis.
Determining Total Extractable Phenolics
Folin-Ciocalteu Method
This method quantifies the total amount of phenolic compounds--both tannins and lower
molecular weight compounds--that can be extracted by organic solvents. For this procedure,
use the same acetone-extracts used to measure extractable condensed tannins in the total
condensed tannin procedure. This procedure uses the Folin-Ciocalteu reagent. It is considered
superior to the older Folin-Dennis reagent in that it is less likely to react with non-phenolic
reducing agents such as ascorbic acid (Julkunen-Titto 1985).
The Folin-Ciocalteu reagent contains sodium molybdate and sodium tungstate. These
compounds react with the hydroxyl group (OH-) in phenolic compounds. The products of these
reactions form a blue color which can be measured by spectrophotometer. The exact shade of
blue that is produced depends upon both the concentration of phenolics in an extract and the
position of the hydroxyl groups on the phenolic molecules (Julkunen-Titto 1985).
I. Preparation of Folin-Ciocalteu Reagent and Sodium Carbonate Buffer Solutions
A.) To prepare the Folin-Ciocalteu reagent and make mixture of 10% purchased Folin-Ciocalteu
concentrate (Sigma F-9252) and 90% distilled water. It is important to a mix a new batch each
day. If multiple “runs” are to be made throughout the day, try to minimize the amount of light
exposure the Folin-Ciocalteu reagent receives.
B.) To prepare the sodium carbonate buffer dissolve 75 grams of Na2CO3*H2O (not the
anhydrous sodium carbonate) in one liter of distilled water. To minimize the time needed to
dissolve the sodium carbonate use a heated magnetic stirring apparatus at low heat. It may
take over an hour for sodium carbonate to completely dissolve. Once mixed the sodium
carbonate buffer solution is quite stable and be kept for several weeks.
II. Performing the Procedure
A.) The standard curve can be constructed with the condensed tannin stock solution used in the
Total Condensed Tannin procedure. Alternatively the curve can be constructed using a stock
solution made by dissolving 100 mg of gallic acid (Sigma G-7384) in 100 ml of distilled water. It
will take about 45 minutes on a magnetic stirring apparatus to dissolve the gallic acid. A new
gallic acid stock solution needs to be made each day. My research showed that the standard
curve obtained with purified sericea lespedeza tannin was very similar to that obtained with
gallic acid.
B.) Construct the standard curve by pipetting 20, 50, 100, 150 and 200 ul aliquots into 25 x 150
ml test tubes.
C.) Pipette 100 ul of acetone extract into the 25 x 150 ml test tubes. Produce 3 tubes for each
subsample, a 3x3 subsampling regime, 3 subsamples per sample and 3 tubes read from each
subsample.
D.) Pipette 5 ml of Folin-Ciocalteu reagent into each test tube, including one blank tube. Vortex
mix each tube, then pipette 4 ml of sodium carbonate buffer solution into each tube. There are
a few important notes on this step. First, the Folin reagent must be added 2-3 minutes before
the sodium carbonate solution or an incomplete reaction will result. Second, it is important to
run similar sized batches so that the time interval between the time the Folin reagent is added
and the buffer is added is relatively consistent across samples.
E.) Let the tubes react at room temperature for 2 hours.
F.) Set the spectrophotometer at 675 nm with filter appropriate for 675 nm wavelength. Blank
spectrophotometer with the blank tube and read the standards and samples. It is important to
run small enough batches that all of the tubes can be read within an hour time frame.
G.) Calculate percent total extractable phenolics as with the extractable condensed tannin
procedure.
MSDS-TANNIN ANALYSES
1. Acetone
Hazards Identification
Potential Acute Health Effects: Hazardous in case of skin contact (irritant), of eye contact
(irritant), of ingestion, of inhalation. Slightly hazardous in case of skin contact (permeator).
Potential Chronic Health Effects:
CARCINOGENIC EFFECTS: A4 (Not classifiable for human or animal.) by ACGIH. MUTAGENIC
EFFECTS: Not available.
TERATOGENIC EFFECTS: Not available. DEVELOPMENTAL TOXICITY: Classified Reproductive
system/toxin/female, Reproductive system/toxin/male [SUSPECTED]. The substance is toxic to
central nervous system (CNS). The substance may be toxic to kidneys, the reproductive system,
liver, skin. Repeated or prolonged exposure to the substance can produce target organs
damage.
First Aid Measures
Eye Contact:
Check for and remove any contact lenses. Immediately flush eyes with running water for at
least 15 minutes, keeping eyelids open. Cold water may be used. Get medical attention.
Skin Contact:
In case of contact, immediately flush skin with plenty of water. Cover the irritated skin with an
emollient. Remove contaminated clothing and shoes. Cold water may be used. Wash clothing
before reuse. Thoroughly clean shoes before reuse. Get medical attention.
Serious Skin Contact:
Wash with a disinfectant soap and cover the contaminated skin with an anti-bacterial cream.
Seek medical attention.
Inhalation:
If inhaled, remove to fresh air. If not breathing, give artificial respiration. If breathing is difficult,
give oxygen. Get medical attention if symptoms appear.
Serious Inhalation:
Evacuate the victim to a safe area as soon as possible. Loosen tight clothing such as a collar, tie,
belt or waistband. If breathing is difficult, administer oxygen. If the victim is not breathing,
perform mouth-to-mouth resuscitation. Seek medical attention.
Ingestion:
Do NOT induce vomiting unless directed to do so by medical personnel. Never give anything by
mouth to an unconscious person. Loosen tight clothing such as a collar, tie, belt or waistband.
Get medical attention if symptoms appear.
Serious Ingestion: Not available.
Flammability of the Product: Flammable.
Auto-Ignition Temperature: 465°C (869°F)
Flash Points: CLOSED CUP: -20°C (-4°F). OPEN CUP: -9°C (15.8°F) (Cleveland).
Flammable Limits: LOWER: 2.6% UPPER: 12.8%
Products of Combustion: These products are carbon oxides (CO, CO2).
Fire Hazards in Presence of Various Substances: Highly flammable in presence of open flames
and sparks, of heat.
Explosion Hazards in Presence of Various Substances:
Risks of explosion of the product in presence of mechanical impact: Not available. Slightly
explosive in presence of open flames and sparks, of oxidizing materials, of acids.
Fire Fighting Media and Instructions:
Flammable liquid, soluble or dispersed in water. SMALL FIRE: Use DRY chemical powder. LARGE
FIRE: Use alcohol foam, water spray or fog.
Special Remarks on Fire Hazards: Vapor may travel considerable distance to source of ignition
and flash back.
Special Remarks on Explosion Hazards:
Forms explosive mixtures with hydrogen peroxide, acetic acid, nitric acid, nitric acid + sulfuric
acid, chromic anydride, chromyl chloride, nitrosyl chloride, hexachloromelamine, nitrosyl
perchlorate, nitryl perchlorate, permonosulfuric acid, thiodiglycol + hydrogen peroxide,
potassium ter-butoxide, sulfur dichloride, 1-methyl-1,3-butadiene, bromoform, carbon, air,
chloroform, thitriazylperchlorate.
Spills
Small Spill:
Dilute with water and mop up, or absorb with an inert dry material and place in an appropriate
waste disposal container.
Large Spill:
Flammable liquid. Keep away from heat. Keep away from sources of ignition. Stop leak if
without risk. Absorb with DRY earth, sand or other non-combustible material. Do not touch
spilled material. Prevent entry into sewers, basements or confined areas; dike if needed. Be
careful that the product is not present at a concentration level above TLV. Check TLV on the
MSDS and with local authorities.
Precautions:
Keep locked up. Keep away from heat. Keep away from sources of ignition. Ground all
equipment containing material. Do not ingest. Do not breathe gas/fumes/ vapor/spray. Wear
suitable protective clothing. In case of insufficient ventilation, wear suitable respiratory
equipment. If ingested, seek medical advice immediately and show the container or the label.
Avoid contact with skin and eyes. Keep away from incompatibles such as oxidizing agents,
reducing agents, acids, alkalis.
Storage:
Store in a segregated and approved area (flammables area) . Keep container in a cool, wellventilated area. Keep container tightly closed and sealed until ready for use. Keep away from
direct sunlight and heat and avoid all possible sources of ignition (spark or flame).
Engineering Controls:
Provide exhaust ventilation or other engineering controls to keep the airborne concentrations
of vapors below their respective threshold limit value. Ensure that eyewash stations and safety
showers are proximal to the work-station location.
Personal Protection:
Splash goggles. Lab coat. Vapor respirator. Be sure to use an approved/certified respirator or
equivalent. Gloves.
Personal Protection in Case of a Large Spill:
Splash goggles. Full suit. Vapor respirator. Boots. Gloves. A self contained breathing apparatus
should be used to avoid inhalation of the product. Suggested protective clothing might not be
sufficient; consult a specialist BEFORE handling this product.
Exposure Limits:
TWA: 500 STEL: 750 (ppm) from ACGIH (TLV) [United States] TWA: 750 STEL: 1000 (ppm) from
OSHA (PEL) [United
States] TWA: 500 STEL: 1000 [Austalia] TWA: 1185 STEL: 2375 (mg/m3) [Australia] TWA: 750
STEL: 1500 (ppm) [United
Kingdom (UK)] TWA: 1810 STEL: 3620 (mg/m3) [United Kingdom (UK)] TWA: 1800 STEL: 2400
from OSHA (PEL) [United States]Consult local authorities for acceptable exposure limits.
2. Methanol
Potential Acute Health Effects:
Hazardous in case of skin contact (irritant), of eye contact (irritant), of ingestion, of inhalation.
Slightly hazardous in case of skin contact (permeator). Severe over-exposure can result in
death.
Potential Chronic Health Effects:
Slightly hazardous in case of skin contact (sensitizer). CARCINOGENIC EFFECTS: Not available.
MUTAGENIC EFFECTS: Mutagenic for mammalian somatic cells. Mutagenic for bacteria and/or
yeast. TERATOGENIC EFFECTS: Classified
POSSIBLE for human.
DEVELOPMENTAL TOXICITY: Not available. The substance is toxic to eyes. The substance may be
toxic to blood, kidneys, liver, brain, peripheral nervous system, upper respiratory tract, skin,
central nervous system (CNS), optic nerve. Repeated or prolonged exposure to the substance
can produce target organs damage. Repeated exposure to a highly toxic material may produce
general deterioration of health by an accumulation in one or many human organs.
First Aid Measures
Eye Contact:
Check for and remove any contact lenses. Immediately flush eyes with running water for at
least 15 minutes, keeping eyelids open. Cold water may be used. Get medical attention.
Skin Contact:
In case of contact, immediately flush skin with plenty of water for at least 15 minutes while
removing contaminated clothing and shoes. Cover the irritated skin with an emollient. Cold
water may be used. Wash clothing before reuse. Thoroughly clean shoes before reuse. Get
medical attention immediately.
Serious Skin Contact:
Wash with a disinfectant soap and cover the contaminated skin with an anti-bacterial cream.
Seek immediate medical attention.
Inhalation:
If inhaled, remove to fresh air. If not breathing, give artificial respiration. If breathing is difficult,
give oxygen. Get medical attention immediately.
Serious Inhalation:
Evacuate the victim to a safe area as soon as possible. Loosen tight clothing such as a collar, tie,
belt or waistband. If breathing is difficult, administer oxygen. If the victim is not breathing,
perform mouth-to-mouth resuscitation. WARNING: It may be hazardous to the person
providing aid to give mouth-to-mouth resuscitation when the inhaled material is toxic,
infectious or corrosive. Seek immediate medical attention.
Ingestion:
If swallowed, do not induce vomiting unless directed to do so by medical personnel. Never give
anything by mouth to an unconscious person. Loosen tight clothing such as a collar, tie, belt or
waistband. Get medical attention immediately.
Serious Ingestion: Not available.
Fire and Explosion Data
Flammability of the Product: Flammable.
Auto-Ignition Temperature: 464°C (867.2°F)
Flash Points: CLOSED CUP: 12°C (53.6°F). OPEN CUP: 16°C (60.8°F).
Flammable Limits: LOWER: 6% UPPER: 36.5%
Products of Combustion: These products are carbon oxides (CO, CO2).
Fire Hazards in Presence of Various Substances:
Highly flammable in presence of open flames and sparks, of heat. Non-flammable in presence
of shocks.
Explosion Hazards in Presence of Various Substances:
Risks of explosion of the product in presence of mechanical impact: Not available. Explosive in
presence of open flames and sparks, of heat.
Fire Fighting Media and Instructions:
Flammable liquid, soluble or dispersed in water. SMALL FIRE: Use DRY chemical powder. LARGE
FIRE: Use alcohol foam, water spray or fog.
Special Remarks on Fire Hazards:
Explosive in the form of vapor when exposed to heat or flame. Vapor may travel considerable
distance to source of ignition and flash back. When heated to decomposition, it emits acrid
smoke and irritating fumes. CAUTION: MAY BURN WITH NEAR INVISIBLE FLAME
Special Remarks on Explosion Hazards:
Forms an explosive mixture with air due to its low flash point. Explosive when mixed with
Choroform + sodium methoxide and diethyl zinc. It boils violently and explodes.
Spills
Small Spill:
Dilute with water and mop up, or absorb with an inert dry material and place in an appropriate
waste disposal container.
Large Spill:
Flammable liquid. Poisonous liquid. Keep away from heat. Keep away from sources of ignition.
Stop leak if without risk.
Absorb with DRY earth, sand or other non-combustible material. Do not get water inside
container. Do not touch spilled material. Use water spray to reduce vapors. Prevent entry into
sewers, basements or confined areas; dike if needed. Call for assistance on disposal. Be careful
that the product is not present at a concentration level above TLV. Check TLV on the
MSDS and with local authorities.
Handling and Storage
Precautions:
Keep locked up. Keep away from heat. Keep away from sources of ignition. Ground all
equipment containing material. Do not ingest. Do not breathe gas/fumes/ vapor/spray. Wear
suitable protective clothing. In case of insufficient ventilation, wear suitable respiratory
equipment. If ingested, seek medical advice immediately and show the container or the label.
Avoid contact with skin and eyes. Keep away from incompatibles such as oxidizing agents,
metals, acids.
Storage:
Store in a segregated and approved area. Keep container in a cool, well-ventilated area. Keep
container tightly closed and sealed until ready for use. Avoid all possible sources of ignition
(spark or flame).
Exposure
Engineering Controls:
Provide exhaust ventilation or other engineering controls to keep the airborne concentrations
of vapors below their respective threshold limit value. Ensure that eyewash stations and safety
showers are proximal to the work-station location.
Personal Protection:
Splash goggles. Lab coat. Vapor respirator. Be sure to use an approved/certified respirator or
equivalent. Gloves.
Personal Protection in Case of a Large Spill:
Splash goggles. Full suit. Vapor respirator. Boots. Gloves. A self-contained breathing apparatus
should be used to avoid inhalation of the product. Suggested protective clothing might not be
sufficient; consult a specialist BEFORE handling this product.
Exposure Limits:
TWA: 200 from OSHA (PEL) [United States] TWA: 200 STEL: 250 (ppm) from ACGIH (TLV) [United
States] [1999] STEL: 250 from NIOSH [United States] TWA: 200 STEL: 250 (ppm) from NIOSH
SKIN TWA: 200 STEL: 250 (ppm) [Canada] Consult local authorities for acceptable exposure
limits.
3-Butanol
Hazards Identification
Potential Acute Health Effects:
Very hazardous in case of skin contact (irritant, permeator), of eye contact (irritant), of
ingestion, of inhalation. Slightly hazardous in case of skin contact (sensitizer). Inflammation of
the eye is characterized by redness, watering, and itching. Skin inflammation is characterized by
itching, scaling, reddening, or, occasionally, blistering.
Potential Chronic Health Effects:
CARCINOGENIC EFFECTS: Not available. MUTAGENIC EFFECTS: Not available. TERATOGENIC
EFFECTS: Not available. DEVELOPMENTAL TOXICITY: Not available. Repeated or prolonged
exposure is not known to aggravate medical condition.
First Aid Measures
Eye Contact:
Check for and remove any contact lenses. Immediately flush eyes with running water for at
least 15 minutes, keeping eyelids open. Cold water may be used. Get medical attention
immediately.
Skin Contact:
In case of contact, immediately flush skin with plenty of water for at least 15 minutes while
removing contaminated clothing and shoes. Cover the irritated skin with an emollient. Cold
water may be used. Wash clothing before reuse. Thoroughly clean shoes before reuse. Get
medical attention immediately.
Serious Skin Contact:
Wash with a disinfectant soap and cover the contaminated skin with an anti-bacterial cream.
Seek immediate medical attention.
Inhalation:
If inhaled, remove to fresh air. If not breathing, give artificial respiration. If breathing is difficult,
give oxygen. Get medical attention if symptoms appear.
Serious Inhalation:
Evacuate the victim to a safe area as soon as possible. Loosen tight clothing such as a collar, tie,
belt or waistband. If breathing is difficult, administer oxygen. If the victim is not breathing,
perform mouth-to-mouth resuscitation. Seek medical attention.
Ingestion:
Do NOT induce vomiting unless directed to do so by medical personnel. Never give anything by
mouth to an unconscious person. If large quantities of this material are swallowed, call a
physician immediately. Loosen tight clothing such as a collar, tie, belt or waistband.
Serious Ingestion: Not available.
Flammability
Flammability of the Product: Flammable.
Auto-Ignition Temperature: 343°C (649.4°F)
Flash Points: CLOSED CUP: 28.9°C (84°F). OPEN CUP: 36.1°C (97°F) (Cleveland).
Flammable Limits: LOWER: 1.4% UPPER: 11.2%
Products of Combustion: These products are carbon oxides (CO, CO2).
Fire Hazards in Presence of Various Substances:
Highly flammable in presence of open flames and sparks. Flammable in presence of heat, of
oxidizing materials, of reducing materials, of combustible materials.
Explosion Hazards in Presence of Various Substances:
Risks of explosion of the product in presence of mechanical impact: Not available. Risks of
explosion of the product in presence of static discharge: Not available.
Fire Fighting Media and Instructions:
Flammable liquid, soluble or dispersed in water. SMALL FIRE: Use DRY chemical powder. LARGE
FIRE: Use alcohol foam, water spray or fog. Cool containing vessels with water jet in order to
prevent pressure build-up, autoignition or explosion.
Special Remarks on Fire Hazards: May form explosive mixtures with air. CAUTION: MAY BURN
WITH NEAR INVISIBLE
FLAME
Small Spill:
Dilute with water and mop up, or absorb with an inert dry material and place in an appropriate
waste disposal container.
Large Spill:
Flammable liquid. Keep away from heat. Keep away from sources of ignition. Stop leak if
without risk. Absorb with DRY earth, sand or other non-combustible material. Do not touch
spilled material. Prevent entry into sewers, basements or confined areas; dike if needed. Be
careful that the product is not present at a concentration level above TLV. Check TLV on the
MSDS and with local authorities.
Handling and Storage
Precautions:
Keep away from heat. Keep away from sources of ignition. Ground all equipment containing
material. Do not ingest. Do not breathe gas/fumes/ vapor/spray. In case of insufficient
ventilation, wear suitable respiratory equipment. If ingested, seek medical advice immediately
and show the container or the label. Avoid contact with skin and eyes. Keep away from
incompatibles such as oxidizing agents, reducing agents.
Storage:
Store in a segregated and approved area. Keep container in a cool, well-ventilated area. Keep
container tightly closed and sealed until ready for use. Avoid all possible sources of ignition
(spark or flame).
Exposure Controls
Engineering Controls:
Provide exhaust ventilation or other engineering controls to keep the airborne concentrations
of vapors below their respective threshold limit value. Ensure that eyewash stations and safety
showers are proximal to the work-station location.
Personal Protection:
Splash goggles. Lab coat. Vapor respirator. Be sure to use an approved/certified respirator or
equivalent. Gloves.
Personal Protection in Case of a Large Spill:
Splash goggles. Full suit. Vapor respirator. Boots. Gloves. A self contained breathing apparatus
should be used to avoid inhalation of the product. Suggested protective clothing might not be
sufficient; consult a specialist BEFORE handling this product.
Exposure Limits:
CEIL: 152 (mg/m3) Consult local authorities for acceptable exposure limits.
4. Hydrochloric Acid
Hazards Identification
Potential Acute Health Effects:
Very hazardous in case of skin contact (corrosive, irritant, permeator), of eye contact (irritant,
corrosive), of ingestion. Slightly hazardous in case of inhalation (lung sensitizer). Non-corrosive
for lungs. Liquid or spray mist may produce tissue damage particularly on mucous membranes
of eyes, mouth and respiratory tract. Skin contact may produce burns. Inhalation of the spray
mist may produce severe irritation of respiratory tract, characterized by coughing, choking, or
shortness of breath.
Severe over-exposure can result in death. Inflammation of the eye is characterized by redness,
watering, and itching. Skin inflammation is characterized by itching, scaling, reddening, or,
occasionally, blistering.
Potential Chronic Health Effects:
Slightly hazardous in case of skin contact (sensitizer). CARCINOGENIC EFFECTS: Classified 3 (Not
classifiable for human.) by IARC [Hydrochloric acid]. MUTAGENIC EFFECTS: Not available.
TERATOGENIC EFFECTS: Not available.
DEVELOPMENTAL TOXICITY: Not available. The substance may be toxic to kidneys, liver, mucous
membranes, upper respiratory tract, skin, eyes, Circulatory System, teeth. Repeated or
prolonged exposure to the substance can produce target organs damage. Repeated or
prolonged contact with spray mist may produce chronic eye irritation and severe skin irritation.
Repeated or prolonged exposure to spray mist may produce respiratory tract irritation leading
to frequent attacks of bronchial infection. Repeated exposure to a highly toxic material may
produce general deterioration of health by an accumulation in one or many human organs.
First Aid Measures
Eye Contact:
Check for and remove any contact lenses. In case of contact, immediately flush eyes with plenty
of water for at least 15 minutes. Cold water may be used. Get medical attention immediately.
Skin Contact:
In case of contact, immediately flush skin with plenty of water for at least 15 minutes while
removing contaminated clothing and shoes. Cover the irritated skin with an emollient. Cold
water may be used. Wash clothing before reuse. Thoroughly clean shoes before reuse. Get
medical attention immediately.
Serious Skin Contact:
Wash with a disinfectant soap and cover the contaminated skin with an anti-bacterial cream.
Seek immediate medical attention.
Inhalation:
If inhaled, remove to fresh air. If not breathing, give artificial respiration. If breathing is difficult,
give oxygen. Get medical attention immediately.
Serious Inhalation:
Evacuate the victim to a safe area as soon as possible. Loosen tight clothing such as a collar, tie,
belt or waistband. If breathing is difficult, administer oxygen. If the victim is not breathing,
perform mouth-to-mouth resuscitation. WARNING: It may be hazardous to the person
providing aid to give mouth-to-mouth resuscitation when the inhaled material is toxic,
infectious or corrosive. Seek immediate medical attention.
Ingestion:
If swallowed, do not induce vomiting unless directed to do so by medical personnel. Never give
anything by mouth to an unconscious person. Loosen tight clothing such as a collar, tie, belt or
waistband. Get medical attention immediately.
Serious Ingestion: Not available.
Fire and Explosion Data
Flammability of the Product: Non-flammable.
Auto-Ignition Temperature: Not applicable.
Flash Points: Not applicable.
Flammable Limits: Not applicable.
Products of Combustion: Not available.
Fire Hazards in Presence of Various Substances: of metals
Explosion Hazards in Presence of Various Substances: Non-explosive in presence of open
flames and sparks, of shocks.
Fire Fighting Media and Instructions: Not applicable.
Special Remarks on Fire Hazards:
Non-combustible. Calcium carbide reacts with hydrogen chloride gas with incandescence.
Uranium phosphide reacts with hydrochloric acid to release spontaneously flammable
phosphine. Rubidium acetylene carbides burns with slightly warm hydrochloric acid. Lithium
silicide in contact with hydrogen chloride becomes incandescent. When dilute hydrochloric acid
is used, gas spontaneously flammable in air is evolved. Magnesium boride treated with
concentrated hydrochloric acid produces spontaneously flammble gas. Cesium acetylene
carbide burns hydrogen chloride gas. Cesium carbide ignites in contact with hydrochloric acid
unless acid is dilute. Reacts with most metals to produce flammable Hydrodgen gas.
Special Remarks on Explosion Hazards:
Hydrogen chloride in contact with the following can cause an explosion, ignition on contact, or
other violent/vigorous reaction:
Acetic anhydride AgClO + CCl4 Alcohols + hydrogen cyanide, Aluminum Aluminum-titanium
alloys (with HCl vapor), 2-Amino ethanol, Ammonium hydroxide, Calcium carbide Ca3P2
Chlorine + dinitroanilines (evolves gas), Chlorosulfonic acid Cesium carbide Cesium acetylene
carbide, 1,1-Difluoroethylene Ethylene diamine Ethylene imine, Fluorine, HClO4 Hexalithium
disilicide H2SO4 Metal acetylides or carbides, Magnesium boride, Mercuric sulfate, Oleum,
Potassium permanganate, beta-Propiolactone Propylene oxide Rubidium carbide, Rubidium,
acetylene carbide Sodium (with aqueous HCl), Sodium hydroxide Sodium tetraselenium,
Sulfonic acid, Tetraselenium tetranitride, U3P4 , Vinyl acetate. Silver perchlorate with carbon
tetrachloride in the presence of hydrochloric acid produces trichloromethyl perchlorate which
detonates at 40 deg. C.
Spills
Small Spill:
Dilute with water and mop up, or absorb with an inert dry material and place in an appropriate
waste disposal container. If necessary: Neutralize the residue with a dilute solution of sodium
carbonate.
Large Spill:
Corrosive liquid. Poisonous liquid. Stop leak if without risk. Absorb with DRY earth, sand or
other non-combustible material. Do not get water inside container. Do not touch spilled
material. Use water spray curtain to divert vapor drift. Use water spray to reduce vapors.
Prevent entry into sewers, basements or confined areas; dike if needed. Call for assistance on
disposal. Neutralize the residue with a dilute solution of sodium carbonate. Be careful that the
product is not present at a concentration level above TLV. Check TLV on the MSDS and with
local authorities.
Handling and Storage
Precautions:
Keep locked up. Keep container dry. Do not ingest. Do not breathe gas/fumes/ vapor/spray.
Never add water to this product. In case of insufficient ventilation, wear suitable respiratory
equipment. If ingested, seek medical advice immediately and show the container or the label.
Avoid contact with skin and eyes. Keep away from incompatibles such as oxidizing agents,
organic materials, metals, alkalis, moisture. May corrode metallic surfaces. Store in a metallic or
coated fiberboard drum using a strong polyethylene inner package.
Storage: Keep container tightly closed. Keep container in a cool, well-ventilated area.
Exposure Controls
Engineering Controls:
Provide exhaust ventilation or other engineering controls to keep the airborne concentrations
of vapors below their respective threshold limit value. Ensure that eyewash stations and safety
showers are proximal to the work-station location.
Personal Protection:
Face shield. Full suit. Vapor respirator. Be sure to use an approved/certified respirator or
equivalent. Gloves. Boots.
Personal Protection in Case of a Large Spill:
Splash goggles. Full suit. Vapor respirator. Boots. Gloves. A self-contained breathing apparatus
should be used to avoid inhalation of the product. Suggested protective clothing might not be
sufficient; consult a specialist BEFORE handling this product.
Exposure Limits:
CEIL: 5 (ppm) from OSHA (PEL) [United States] CEIL: 7 (mg/m3) from OSHA (PEL) [United States]
CEIL: 5 from NIOSH
CEIL: 7 (mg/m3) from NIOSH TWA: 1 STEL: 5 (ppm) [United Kingdom (UK)] TWA: 2 STEL: 8
(mg/m3) [United Kingdom
(UK)]Consult local authorities for acceptable exposure limits.
5. MSDS Folin-Ciocalteau Reagent
Hazards Identification
Potential Acute Health Effects:
Very hazardous in case of skin contact (irritant), of eye contact (irritant), of ingestion, .
Hazardous in case of skin contact (corrosive, permeator), of eye contact (corrosive). Liquid or
spray mist may produce tissue damage particularly on mucous membranes of eyes, mouth and
respiratory tract. Skin contact may produce burns. Inhalation of the spray mist may produce
severe irritation of respiratory tract, characterized by coughing, choking, or shortness of breath.
Severe over-exposure can result in death. Inflammation of the eye is characterized by redness,
watering, and itching. Skin inflammation is characterized by itching, scaling, reddening, or,
occasionally, blistering.
Potential Chronic Health Effects:
CARCINOGENIC EFFECTS: Classified 4 (No evidence.) by NTP, None. by OSHA, None. by NIOSH
[Bromine]. Classified 3 (Not classifiable for human.) by IARC [Hydrogen chloride]. MUTAGENIC
EFFECTS: Mutagenic for bacteria and/or yeast.
[Lithium sulfate monohydrate]. TERATOGENIC EFFECTS: Not available. DEVELOPMENTAL
TOXICITY: Not available. The substance is toxic to mucous membranes. The substance may be
toxic to kidneys, the nervous system, liver, cardiovascular system, upper respiratory tract, skin,
eyes, central nervous system (CNS), teeth, thyroid. Repeated or prolonged exposure to the
substance can produce target organs damage. Repeated or prolonged contact with spray mist
may produce chronic eye irritation and severe skin irritation. Repeated or prolonged exposure
to spray mist may produce respiratory tract irritation leading to frequent attacks of bronchial
infection. Repeated exposure to a highly toxic material may produce general deterioration of
health by an accumulation in one or many human organs.
First Aid Measures
Eye Contact:
Check for and remove any contact lenses. In case of contact, immediately flush eyes with plenty
of water for at least 15 minutes. Cold water may be used. Get medical attention immediately.
Skin Contact:
In case of contact, immediately flush skin with plenty of water for at least 15 minutes while
removing contaminated clothing and shoes. Cover the irritated skin with an emollient. Cold
water may be used. Wash clothing before reuse. Thoroughly clean shoes before reuse. Get
medical attention immediately.
Serious Skin Contact:
Wash with a disinfectant soap and cover the contaminated skin with an anti-bacterial cream.
Seek immediate medical attention.
Inhalation:
If inhaled, remove to fresh air. If not breathing, give artificial respiration. If breathing is difficult,
give oxygen. Get medical attention immediately.
Serious Inhalation:
Evacuate the victim to a safe area as soon as possible. Loosen tight clothing such as a collar, tie,
belt or waistband.
WARNING: It may be hazardous to the person providing aid to give mouth-to-mouth
resuscitation when the inhaled material is toxic, infectious or corrosive. Seek immediate
medical attention.
Ingestion:
Do NOT induce vomiting unless directed to do so by medical personnel. Never give anything by
mouth to an unconscious person. If large quantities of this material are swallowed, call a
physician immediately. Loosen tight clothing such as a collar, tie, belt or waistband.
Serious Ingestion: Not available.
Fire and Explosion data
Flammability of the Product: Non-flammable.
Auto-Ignition Temperature: Not applicable.
Flash Points: Not applicable.
Flammable Limits: Not applicable.
Products of Combustion: Not available.
Fire Hazards in Presence of Various Substances: Not applicable.
Explosion Hazards in Presence of Various Substances: Non-explosive in presence of open
flames and sparks, of shocks.
Fire Fighting Media and Instructions: Not applicable.
Special Remarks on Fire Hazards: Not available.
Special Remarks on Explosion Hazards: Mixtures with nitromethane are explosive (Phosphoric
Acid)
Spills
Small Spill:
Dilute with water and mop up, or absorb with an inert dry material and place in an appropriate
waste disposal container. If necessary: Neutralize the residue with a dilute solution of sodium
carbonate.
Large Spill:
Corrosive liquid. Poisonous liquid. Stop leak if without risk. Absorb with DRY earth, sand or
other non-combustible material. Do not get water inside container. Do not touch spilled
material. Use water spray curtain to divert vapor drift. Use water spray to reduce vapors.
Prevent entry into sewers, basements or confined areas; dike if needed. Call for assistance on
disposal. Neutralize the residue with a dilute solution of sodium carbonate. Be careful that the
product is not present at a concentration level above TLV. Check TLV on the MSDS and with
local authorities.
Handling and Storage
Precautions:
Keep locked up. Keep container dry. Do not ingest. Do not breathe gas/fumes/ vapor/spray.
Never add water to this product.
In case of insufficient ventilation, wear suitable respiratory equipment. If ingested, seek medical
advice immediately and show the container or the label. Avoid contact with skin and eyes. May
corrode metallic surfaces. Store in a metallic or coated fiberboard drum using a strong
polyethylene inner package.
Storage: Keep container tightly closed. Keep container in a cool, well-ventilated area.
Engineering Controls:
Provide exhaust ventilation or other engineering controls to keep the airborne concentrations
of vapors below their respective threshold limit value. Ensure that eyewash stations and safety
showers are proximal to the work-station location.
Personal Protection:
Face shield. Full suit. Vapor respirator. Be sure to use an approved/certified respirator or
equivalent. Gloves. Boots.
Personal Protection in Case of a Large Spill:
Splash goggles. Full suit. Vapor respirator. Boots. Gloves. A self contained breathing apparatus
should be used to avoid inhalation of the product. Suggested protective clothing might not be
sufficient; consult a specialist BEFORE handling this product.
Exposure
Exposure Limits:
Sodium molybdate (VI) dihydrate TWA: 5 (mg/m3) from ACGIH (TLV) [United States] Inhalation
Respirable. TWA: 5 (mg/m3) from OSHA (PEL) [United States] Inhalation Respirable. TWA: 15
(mg/m3) from OSHA (PEL) [United States] Inhalation Total.
Bromine TWA: 0.66 STEL: 1.3 (mg/m3) from ACGIH (TLV) [United States] TWA: 0.1 STEL: 0.2
(ppm) from ACGIH (TLV) [United States] TWA: 0.1 from OSHA (PEL) [United States] TWA: 0.7
(mg/m3) from OSHA (PEL) [United States] TWA: 0.66
STEL: 2 (mg/m3) [United Kingdom (UK)] TWA: 1 STEL: 0.3 (ppm) [United Kingdom (UK)]
Hydrogen chloride STEL: 7.5 (mg/ m3) from ACGIH (TLV) [United States] STEL: 5 (ppm) from
ACGIH (TLV) [United States] CEIL: 5 (ppm) from NIOSH CEIL:
7.5 (mg/m3) from NIOSH CEIL: 5 (ppm) from OSHA (PEL) [United States] CEIL: 7 (mg/m3) from
OSHA (PEL) [United States] Phosphoric Acid TWA: 1 STEL: 3 (mg/m3) from ACGIH (TLV) [United
States] TWA: 1 STEL: 3 (mg/m3) from OSHA (PEL) [United States] TWA: 1 STEL: 3 (mg/m3) from
NIOSH TWA: 1 STEL: 3 (mg/m3) [Mexico] Sodium tungstate dihydrate TWA: 5 STEL: 10 (mg/m3)
from ACGIH (TLV) [United States] Bromine33
Standard Operating Procedure – Saponin Analyses
Section 1 - Personal Protective Equipment
1. Lab Coat or Lab Gown
2. Hair tied back if long
3. Proper enclosed footwear
4. Safety glasses or goggles
5. Nitrile or latex gloves
SOP for Saponin Analysis in Alfalfa
Collecting & preparing plant samples:
1. Clip about 50-100 grams of vegetative material and place in cooler with dry or regular
ice until they can be placed in freezer.
2. Freeze-dry plant samples.
3. Grind them in a Wiley mill to pass through a 1mm screen.
4.
Extracting Saponins to create standard:
*note: extraction of saponins for the standard requires 25-50 grams of dried ground material,
depending on how much pure saponin desired (~25 grams plant material = 0.2111 grams
saponin).
STEP A: Soxhlet Extraction
1. Measure 25 grams of ground alfalfa and place in paper thimble.
2. Extract with HPLC grade hexane (~ 700mL) using soxhlet method for ~ 30 hours. Discard
hexane.
3. Remove thimble and set out to dry overnight.
4. Continue soxhlet extraction using methanol (700-750mL) for ~ 30 hours. Keep extraction
with methanol solvent.
STEP B: Chloroform Extraction
1. Use a rotary evaporator (roto-vap) to remove excess methanol solvent (~1.5 hours).
2. Add 600mL of 50/50 chloroform/distilled water to extract and decant into a separation
funnel.
3. Drain lower chloroform layer to discard.
4. Add 300 mL of 50/50 chloroform/water, shake and let settle. Drain lower chloroform
layer to discard.
5. Repeat step 4 for a total of 4 chloroform extractions.
6.
STEP C: Butanol Extraction
1. Combine 300mL N-butanol with 100mL distilled water in separation funnel. Shake and
let settle. Drain lower layer of water do discard.
2. Mix 100mL of water-saturated N-butanol from step C1 with aqueous solution from step
B and 1.5 grams sodium chloride in separation funnel.
3. Drain lower polar layer to discard.
4. Add 100mL water-saturated N-butanol to separation funnel with non-polar layer, mix,
let settle and drain lower polar layer to discard.
5. Repeat step 4 for a total of 3 butanol extractions.
6. Roto-vap extract to dryness.
STEP D: Purifying Saponin Extract
1. Add ~ 4mL methanol to flask and sonicate to dissolve all dry residue.
2. Introduce drop-wise into acetone (~ 900mL).
3. Butchner funnel acetone to isolate precipitate. Add precipitate back into flask with ~
2mL methanol, sonicate and drip into fresh acetone (~ 900mL).
4. Repeat step 3 (2X) and after the fourth and final time, allow the precipitate to stand in
acetone for 16 hours.
5. Collect the precipitate by using a butchner funnel and allow it to dry overnight at room
temperature. Yield = .2111 grams purified saponin.
STEP E: Creating Standard Curve
*Note: Any glassware, especially the test tubes and caps, used for the entire procedure must be
washed and rinsed properly to remove all residual soap or it will confound results.
1. Measure 10mg saponin extract and add to 10mL distilled water in 16mL glass centrifuge
test tube.
2. Using a serial dilution create a 1mg/ml, 0.5mg/ml, 0.25mg/ml, 0.125mg/ml, etc… in 7
(exact brand as the first) test tubes.
3. Shake for 2 minutes and let settle for 15-20 minutes. Measure and record foam.
4. Using excel obtain an R² value for the curve. Should be at least 0.97.
Determining saponin % using small-scale extractions:
Method developed by Steve, Dale, and Andrea
STEP A: Methanol Extraction
1. Weigh out 50 mg of dried and ground plant material in triplicates and put into 16ml
screw-cap test tubes.
2. Add 5 mL methanol and vortex for 2 minutes or put on roto-geni for 30 minutes.
3.
4.
5.
6.
Centrifuge for 5 minutes at 2500 rpm.
Pipette or decant supernatant into 20 mL vials.
Repeat steps 2-3 for a total of 2 methanol extractions.
Dry down methanol extractions.
STEP B: Chloroform Extraction
*Note: use tephlon caps and glass pipettes and vials for chloroform extractions.
1. Add 5 mL distilled water and 5 mL chloroform to each sample from step A. Shake until
all dried residue is dissolved.
2. Centrifuge 5 minutes at 2500rpm.
3. Pipette lower chloroform layer out to discard.
4. Add 5ml chloroform to each vial.
5. Repeat steps 2-4 for a total of 4 chloroform extractions.
STEP C: N-Butanol Extraction
1. Combine 150mL N-butanol with 50mL distilled water and 0.75 grams sodium chloride in
separation funnel. Shake and let settle. Drain lower layer of water to discard.
2. After discarding the lower chloroform layer from the 4th chloroform extraction in step B,
add 5ml water-saturated N-butanol (from step C1) to each sample.
3. Shake then centrifuge 5 minutes at 2500rpm.
4. Pipette upper non-polar layer out and put into new and labeled 16mL test tubes.
*Note: these test tubes will be used for the foam test so they must be the same brand used
in creating the standard curve. Also, if one doesn’t have a nitrogen dryer to dry samples
down, this step may use 20mL vials, put in sand bath and improvise blowers with glass
pipettes and tubing. The dry residue will dissolve in water and can then be transferred to a
16mL centrifuge tube for the foam test.
5. Add 5ml water-saturated N-butanol to each sample from step C2. Shake and centrifuge
5 minutes at 2500rpm.
6. Repeat step C4 so there is an accumulation of ~ 15ml of non-polar supernatant in the
new and labeled test tubes.
7. Dry down N-butanol extractions.
STEP D: Foam Test
1. Add 5mL distilled water to each sample. Shake for 2 minutes, let stand for 15-20
minutes then measure and record foam.
2. Calculate the percent of saponins by calculating mg/ml based on the standard curve
equation (for instance, if the 0.25mg/ml from the standard rendered 2.8 cm of foam,
use the number 0.089 by dividing 0.25 by 2.8), multiply by the cm of foam the sample
rendered, multiply by 5, divide by the weight of the sample used in the extraction, lastly
multiply by 100 for a percent by weight saponin content.
*Note: foam may be reduced (if a re-measurement is required) by centrifuging.
1. Hexane
MSDS-SAPONIN ANALYSES
Hazards Identification
Potential Acute Health Effects:
Hazardous in case of skin contact (permeator), of ingestion, of inhalation. Slightly hazardous in
case of skin contact (irritant), of eye contact (irritant).
Potential Chronic Health Effects:
CARCINOGENIC EFFECTS: Not available. MUTAGENIC EFFECTS: Mutagenic for bacteria and/or
yeast. TERATOGENIC EFFECTS: Not available. DEVELOPMENTAL TOXICITY: Not available. The
substance may be toxic to peripheral nervous system, skin, central nervous system (CNS).
Repeated or prolonged exposure to the substance can produce target organs damage.
First Aid Measures
Eye Contact:
Check for and remove any contact lenses. Immediately flush eyes with running water for at
least 15 minutes, keeping eyelids open. Get medical attention if irritation occurs.
Skin Contact: Wash with soap and water. Cover the irritated skin with an emollient. Get
medical attention if irritation develops.
Serious Skin Contact:
Wash with a disinfectant soap and cover the contaminated skin with an anti-bacterial cream.
Seek medical attention.
Inhalation:
If inhaled, remove to fresh air. If not breathing, give artificial respiration. If breathing is difficult,
give oxygen. Get medical attention if symptoms appear.
Serious Inhalation:
Evacuate the victim to a safe area as soon as possible. Loosen tight clothing such as a collar, tie,
belt or waistband. If breathing is difficult, administer oxygen. If the victim is not breathing,
perform mouth-to-mouth resuscitation. Seek medical attention.
Ingestion:
Do NOT induce vomiting unless directed to do so by medical personnel. Never give anything by
mouth to an unconscious person. Loosen tight clothing such as a collar, tie, belt or waistband.
Get medical attention if symptoms appear.
Serious Ingestion: Not available.
Flammability of the Product: Flammable.
Auto-Ignition Temperature: 225°C (437°F)
Flash Points: CLOSED CUP: -22.5°C (-8.5°F). (TAG)
Flammable Limits: LOWER: 1.15% UPPER: 7.5%
Products of Combustion: These products are carbon oxides (CO, CO2).
Fire Hazards in Presence of Various Substances:
Highly flammable in presence of open flames and sparks, of heat. Non-flammable in presence
of shocks.
Explosion Hazards in Presence of Various Substances:
Risks of explosion of the product in presence of mechanical impact: Not available.
Risks of explosion of the product in presence of static discharge: Not available.
Fire and Explosion Data
Fire Fighting Media and Instructions:
Flammable liquid, insoluble in water. SMALL FIRE: Use DRY chemical powder. LARGE FIRE: Use
water spray or fog.
Special Remarks on Fire Hazards:
Extremely flammable liquid and vapor. Vapor may cause flash fire.
Special Remarks on Explosion Hazards: Not available.
Spills
Small Spill: Absorb with an inert material and put the spilled material in an appropriate waste
disposal.
Large Spill:
Flammable liquid, insoluble in water. Keep away from heat. Keep away from sources of ignition.
Stop leak if without risk.
Absorb with DRY earth, sand or other non-combustible material. Do not get water inside
container. Do not touch spilled material. Prevent entry into sewers, basements or confined
areas; dike if needed. Call for assistance on disposal. Be careful that the product is not present
at a concentration level above TLV. Check TLV on the MSDS and with local authorities.
Handling and Storage
Precautions:
Keep locked up. Keep away from heat. Keep away from sources of ignition. Ground all
equipment containing material.
Do not ingest. Do not breathe gas/fumes/ vapor/spray. Avoid contact with skin. Wear suitable
protective clothing. In case of insufficient ventilation, wear suitable respiratory equipment. If
ingested, seek medical advice immediately and show the container or the label. Keep away
from incompatibles such as oxidizing agents.
Storage:
Store in a segregated and approved area. Keep container in a cool, well-ventilated area. Keep
container tightly closed and sealed until ready for use. Avoid all possible sources of ignition
(spark or flame).
Exposure Controls/Personal Protection
Engineering Controls:
Provide exhaust ventilation or other engineering controls to keep the airborne concentrations
of vapors below their respective threshold limit value. Ensure that eyewash stations and safety
showers are proximal to the work-station location.
Personal Protection:
Safety glasses. Lab coat. Vapor respirator. Be sure to use an approved/certified respirator or
equivalent. Gloves (impervious).
Personal Protection in Case of a Large Spill:
Splash goggles. Full suit. Vapor respirator. Boots. Gloves. A self-contained breathing apparatus
should be used to avoid inhalation of the product. Suggested protective clothing might not be
sufficient; consult a specialist BEFORE handling this product.
Exposure Limits:
TWA: 500 (ppm) from OSHA (PEL) [United States] Inhalation TWA: 1800 (mg/m3) from OSHA
(PEL) [United States] Inhalation TWA: 176 (mg/m3) from ACGIH (TLV) [United States] SKIN TWA:
50 (ppm) from ACGIH (TLV) [United States] SKIN TWA: 500 STEL: 1000 (ppm) from ACGIH (TLV)
[United States] Inhalation TWA: 1760 STEL: 3500 (mg/m3) from ACGIH (TLV) [United States]
Inhalation Consult local authorities for acceptable exposure limits.
2. Butanol
Hazards Identification
Potential Acute Health Effects:
Very hazardous in case of skin contact (irritant, permeator), of eye contact (irritant), of
ingestion, of inhalation. Slightly hazardous in case of skin contact (sensitizer). Inflammation of
the eye is characterized by redness, watering, and itching. Skin inflammation is characterized by
itching, scaling, reddening, or, occasionally, blistering.
Potential Chronic Health Effects:
CARCINOGENIC EFFECTS: Not available. MUTAGENIC EFFECTS: Not available. TERATOGENIC
EFFECTS: Not available.
DEVELOPMENTAL TOXICITY: Not available. Repeated or prolonged exposure is not known to
aggravate medical condition.
First Aid Measures
Eye Contact:
Check for and remove any contact lenses. Immediately flush eyes with running water for at
least 15 minutes, keeping eyelids open. Cold water may be used. Get medical attention
immediately.
Skin Contact:
In case of contact, immediately flush skin with plenty of water for at least 15 minutes while
removing contaminated clothing and shoes. Cover the irritated skin with an emollient. Cold
water may be used. Wash clothing before reuse. Thoroughly clean shoes before reuse. Get
medical attention immediately.
Serious Skin Contact:
Wash with a disinfectant soap and cover the contaminated skin with an anti-bacterial cream.
Seek immediate medical attention.
Inhalation:
If inhaled, remove to fresh air. If not breathing, give artificial respiration. If breathing is difficult,
give oxygen. Get medical attention if symptoms appear.
Serious Inhalation:
Evacuate the victim to a safe area as soon as possible. Loosen tight clothing such as a collar, tie,
belt or waistband. If breathing is difficult, administer oxygen. If the victim is not breathing,
perform mouth-to-mouth resuscitation. Seek medical attention.
Ingestion:
Do NOT induce vomiting unless directed to do so by medical personnel. Never give anything by
mouth to an unconscious person. If large quantities of this material are swallowed, call a
physician immediately. Loosen tight clothing such as a collar, tie, belt or waistband.
Serious Ingestion: Not available.
Fire and Explosion Data
Flammability of the Product: Flammable.
Auto-Ignition Temperature: 343°C (649.4°F)
Flash Points: CLOSED CUP: 28.9°C (84°F). OPEN CUP: 36.1°C (97°F) (Cleveland).
Flammable Limits: LOWER: 1.4% UPPER: 11.2%
Products of Combustion: These products are carbon oxides (CO, CO2).
Fire Hazards in Presence of Various Substances:
Highly flammable in presence of open flames and sparks. Flammable in presence of heat, of
oxidizing materials, of reducing materials, of combustible materials.
Explosion Hazards in Presence of Various Substances:
Risks of explosion of the product in presence of mechanical impact: Not available. Risks of
explosion of the product in presence of static discharge: Not available.
Fire Fighting Media and Instructions:
Flammable liquid, soluble or dispersed in water. SMALL FIRE: Use DRY chemical powder. LARGE
FIRE: Use alcohol foam, water spray or fog. Cool containing vessels with water jet in order to
prevent pressure build-up, autoignition or explosion.
Special Remarks on Fire Hazards: May form explosive mixtures with air. CAUTION: MAY BURN
WITH NEAR INVISIBLE FLAME
Special Remarks on Explosion Hazards: Not available.
Spills
Small Spill:
Dilute with water and mop up, or absorb with an inert dry material and place in an appropriate
waste disposal container.
Large Spill:
p. 3
Flammable liquid. Keep away from heat. Keep away from sources of ignition. Stop leak if
without risk. Absorb with DRY earth, sand or other non-combustible material. Do not touch
spilled material. Prevent entry into sewers, basements or confined areas; dike if needed. Be
careful that the product is not present at a concentration level above TLV. Check TLV on the
MSDS and with local authorities.
Handling and storage
Precautions:
Keep away from heat. Keep away from sources of ignition. Ground all equipment containing
material. Do not ingest. Do not breathe gas/fumes/ vapor/spray. In case of insufficient
ventilation, wear suitable respiratory equipment. If ingested, seek medical advice immediately
and show the container or the label. Avoid contact with skin and eyes. Keep away from
incompatibles such as oxidizing agents, reducing agents.
Storage:
Store in a segregated and approved area. Keep container in a cool, well-ventilated area. Keep
container tightly closed and sealed until ready for use. Avoid all possible sources of ignition
(spark or flame).
Exposure Controls
Engineering Controls:
Provide exhaust ventilation or other engineering controls to keep the airborne concentrations
of vapors below their respective threshold limit value. Ensure that eyewash stations and safety
showers are proximal to the work-station location.
Personal Protection:
Splash goggles. Lab coat. Vapor respirator. Be sure to use an approved/certified respirator or
equivalent. Gloves.
Personal Protection in Case of a Large Spill:
Splash goggles. Full suit. Vapor respirator. Boots. Gloves. A self contained breathing apparatus
should be used to avoid inhalation of the product. Suggested protective clothing might not be
sufficient; consult a specialist BEFORE handling this product.
Exposure Limits:
CEIL: 152 (mg/m3) Consult local authorities for acceptable exposure limits.
3. Chloroform
Hazards Identification
Potential Acute Health Effects: Hazardous in case of skin contact (irritant), of eye contact
(irritant), of ingestion, of inhalation.
Slightly hazardous in case of skin contact (permeator).
Potential Chronic Health Effects: CARCINOGENIC EFFECTS: Classified + (Proven.) by NIOSH.
Classified A3 (Proven for animal.) by ACGIH, 2B (Possible for human.) by IARC. Classified 2
(Some evidence.) by NTP. MUTAGENIC EFFECTS: Mutagenic for mammalian somatic cells.
Mutagenic for bacteria and/or yeast. TERATOGENIC EFFECTS: Not available.
DEVELOPMENTAL TOXICITY: Not available. The substance may be toxic to kidneys, liver, heart.
Repeated or prolonged exposure to the substance can produce target organs damage.
First Aid Measures
Eye Contact: Check for and remove any contact lenses. In case of contact, immediately flush
eyes with plenty of water for at least 15 minutes. Cold water may be used. WARM water MUST
be used. Get medical attention.
Skin Contact: In case of contact, immediately flush skin with plenty of water. Cover the irritated
skin with an emollient. Remove contaminated clothing and shoes. Wash clothing before reuse.
Thoroughly clean shoes before reuse. Get medical attention.
Serious Skin Contact: Wash with a disinfectant soap and cover the contaminated skin with an
anti-bacterial cream. Seek immediate medical attention.
Inhalation: If inhaled, remove to fresh air. If not breathing, give artificial respiration. If
breathing is difficult, give oxygen. Get medical attention.
Serious Inhalation: Evacuate the victim to a safe area as soon as possible. Loosen tight clothing
such as a collar, tie, belt or waistband. If breathing is difficult, administer oxygen. If the victim is
not breathing, perform mouth-to-mouth resuscitation.
WARNING: It may be hazardous to the person providing aid to give mouth-to-mouth
resuscitation when the inhaled material is toxic, infectious or corrosive. Seek medical attention.
Ingestion: Do NOT induce vomiting unless directed to do so by medical personnel. Never give
anything by mouth to an unconscious person. If large quantities of this material are swallowed,
call a physician immediately. Loosen tight clothing such as a collar, tie, belt or waistband.
Serious Ingestion: Not available.
Fire and Explosion Data
Flammability of the Product: Non-flammable.
Auto-Ignition Temperature: Not applicable.
Flash Points: Not applicable.
Flammable Limits: Not applicable.
Products of Combustion: Not available.
Fire Hazards in Presence of Various Substances: Not applicable.
Explosion Hazards in Presence of Various Substances: Risks of explosion of the product in
presence of mechanical impact: Not available. Risks of explosion of the product in presence of
static discharge: Not available.
Fire Fighting Media and Instructions: Not applicable.
Special Remarks on Fire Hazards: Not available.
Special Remarks on Explosion Hazards: May explode if it comes in contact with aluminum
powder, lithium, perchlorate, pentoxide, bis(dimethylamino)dimethylstannane, potassium,
potassium-sodium alloy, sodium (or sodium hydroxide or sodium methoxide), and methanol
Spills
Small Spill: Absorb with an inert material and put the spilled material in an appropriate waste
disposal.
Large Spill: Absorb with an inert material and put the spilled material in an appropriate waste
disposal. Be careful that the product is not present at a concentration level above TLV. Check
TLV on the MSDS and with local authorities.
Handling and Storage
Precautions: Do not ingest. Do not breathe gas/fumes/ vapor/spray. Wear suitable protective
clothing. In case of insufficient ventilation, wear suitable respiratory equipment. If ingested,
seek medical advice immediately and show the container or the label. Avoid contact with skin
and eyes. Keep away from incompatibles such as metals, alkalis.
Storage: Keep container tightly closed. Keep container in a cool, well-ventilated area. Sensitive
to light. Store in light-resistant containers.
Exposure Controls/Personal Protection
Engineering Controls: Provide exhaust ventilation or other engineering controls to keep the
airborne concentrations of vapors below their respective threshold limit value. Ensure that
eyewash stations and safety showers are proximal to the workstation location.
Personal Protection: Splash goggles. Lab coat. Vapor respirator. Be sure to use an
approved/certified respirator or equivalent. Gloves.
Personal Protection in Case of a Large Spill: Splash goggles. Full suit. Vapor respirator. Boots.
Gloves. A self-contained breathing apparatus should be used to avoid inhalation of the product.
Suggested protective clothing might not be sufficient; consult a specialist BEFORE handling this
product.
Exposure Limits: TWA: 10 (ppm) [Australia] Inhalation TWA: 2 (ppm) from OSHA (PEL) [United
States] Inhalation STEL: 9.78 (mg/m3) from NIOSH Inhalation STEL: 2 (ppm) from NIOSH
Inhalation TWA: 9.78 (mg/m3) from OSHA (PEL) [United States] Inhalation TWA: 10 (ppm) from
ACGIH (TLV) [United States] [1999] Inhalation TWA: 2 (ppm) [United Kingdom (UK)] Inhalation
TWA: 9.9 (mg/m3) [United Kingdom (UK)] InhalationConsult local authorities for acceptable
exposure limits.
CHEMICAL HIGIENE PLAN -Specific Plan
Laboratory Location: Green Canyon Ecology Center
Supervisor Name: Juan Villalba
Laboratory PI or Supervisor Signoff (required). I certify that I have reviewed and approve the
attached Laboratory-specific Chemical Hygiene Plan with Standard Operating Procedures for
my above laboratory locations.
Signature:________________________________________
Laboratory Worker Training Record: Chemical Hygiene Plan
Principal Investigator/Supervisor: _________________________________
The following lab workers have reviewed and understand the following elements of the
attached Chemical Hygiene Plan for the above PI or supervisor:
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Sec. I: Laboratory-specific Chemical Hygiene Plan with Standard Operating Procedures
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Sec. II: Online Safety Training (https://rgs.usu.edu/ehs/laboratory/)
Name (print)
Signature
Date
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Standard Operating Procedures – Green Canyon Ecology
Center
Standard Operating Procedure – General Care of Livestock
1. Make sure sheep have an ample supply of clean water at all times. Small water troughs
should be cleaned frequently and large troughs should be cleaned at least once a week.
Feed should be clean and mold-free. Feeders should be kept free from fecal material.
Don’t change the type of feed you are feeding abruptly. Abrupt changes in feed can cause
acidosis and/or bloat.
Animals should have access to trace mineralized salt blocks at all times.
Vaccinations. Lambs should be given vaccinated against Clostridium (enterotoxemia) and
tetanus at 30 and 60 days of age.
Lambs should have their feet trimmed and they should receive a drench of antiparasitic drugs in
the spring.
Standard Operating Procedure – Mixing Feed for Rations and Use of Mixer
Operator should be able to demonstrate competent safe operation to supervisor prior to
authorization of use.
Section 1 - Personal Protective Equipment
• Eye protection required
• Long of loose hair must be tied back or contained
• Hearing protection required
• Approved dust mask required
• Work gloves required
• Protective clothing required
• No jewelry, watches, ring, necklaces
• No loose fitting clothing
Section 2 - Potential Hazards + Safety precautions
Crushing hazards when operating the machine. Worker entanglement hazards on moving parts.
Bystander or operator run over. Projectile debris hazards from rotating parts and feed. Noise.
Confined space. No worker should enter the mixer when is not properly checked that the
machine is turned off.
Procedure
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Inspect required personnel protective equipment and replace if required
Pot on all required personnel protective equipment
Start the mixer by pulling the rope attached the engine
Fill and move the lever for mixing operation
After 15-20 minutes (depending on the type of feed) begin discharging the feed material
from mixer to a container
Never climb on mixer during operation.
If an emergency occurs while conducting this task, or there is an equipment malfunction, shut
the equipment off immediately.
Report ant hazardous situation to your supervisor immediately.
Avoid Being Caught in Moving Parts
Feeds should be fed into the mixer safely, to avoid contact with the equipment’s moving parts.
Follow these feeding procedures to keep you safe.
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Insert feeds only when the mixer is at full operating speed.
Keep hands and feet outside of the feed chute.
Let go of feed at least ½ m from the intake and walk away to avoid being hit or dragged
into the mixer by its moving parts.
Safe Fueling Procedures
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Gasoline and other fuels are flammable. Following safe fueling procedures will help
reduce the risk of fuel ignition.
Always shut off the engine and wait at least 5 minutes for the engine to cool before
refueling.
Use only approved fuel containers and store in a well ventilated area, away from direct
sunlight.
Never smoke or have an open flame near fuel.
Touch the fuel nozzle to the chipper/shredder before opening the fuel cap to reduce the
chance that a static spark will ignite the fuel.
Keep the nozzle in contact with the filler neck while fueling.
Replace the cap as soon as you are finished.
Use a funnel or non-spill nozzle when fueling to reduce spillage.
Wipe up all spills immediately, before starting the engine.
Never clean your hands or tools with gasoline. Use a nonflammable solvent instead.
Standard Operating Procedure – Grinding Feed for Rations
Operator should be able to demonstrate competent safe operation to supervisor prior to
authorization of use.
Section 1 - Personal Protective Equipment
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Eye protection required
Long of loose hair must be tied back or contained
Hearing protection required
Approved dust mask required
Work gloves required
Protective clothing required
No jewelry, watches, ring, necklaces
No loose fitting clothing
Close-fitting clothes
Long pants and turdy, non-slip boots
Section 2 - Potential Hazards + Safety precautions
You can be seriously injured or killed if you incorrectly operate a chipper/shredder. The most
commonly reported causes of serious accidents are:
1. Being Caught in Moving Parts
2. Struck by Chipper/Shredder’s Hood
Sometimes you may be tempted to take risky shortcuts. Remember that an accident can leave
you permanently injured or cut your life short. For your safety and the safety of those around
you, do not take unnecessary risks. No deadline is so pressing you can’t take the time to do
your work safely.
Do not operate machinery if you are tired or have taken drugs or alcohol. If you are on
medication, discuss with your doctor or pharmacist if you are capable of safely operating
machinery
Procedure
Place the machine on a level surface that is not concrete, paved or gravel. Operating on these
surfaces can cause thrown material to ricochet and injure or kill the operator or bystanders.
Also keep other workers from falling into the chipper/shredder by ensuring the location of the
machine is not directly in front of or below where other employees are working.
Make sure that there is a tarp below the grinder so you catch all the ground material in the
tarp.
Ensure the hood, enclosing the chipper/shredder’s knives, is closed and latched before
operating. Also, check for loose or missing pins in the hood latch as well as cracked or worn
hinges. If the hood is not properly secured, it could fly off and seriously injure or kill you. The
chipper/shredder should never be operated with the hood open.
Ensure the cutting chamber is free of foreign objects or other debris such as accumulated
woody plant material. Woody plant material and foreign objects can be thrown from the
chipper at high rates of speed.
Fluid leaks can cause fires and breakdowns. Report signs of leaking fluid to your supervisor
immediately.
Move the lever to the “run” position and pull the rope by the engine until the engine starts.
Start the chipper/shredder at the lowest possible speed and listen for any noise or vibration
that could indicate loose or broken parts. If heard, turn off the machine.
Be aware at all times of the location of coworkers. Keep all others out of the area. Check the
operator’s manual for additional procedures.
Begin delivering the feed you want to grind to the intake of the grinder.
You can push the machine in the flat area once there is an accumulation of ground feed
underneath the grinder.
Once you finished grinding, disconnect the power supply by moving the power lever to the off
position. You can tell the machine has stopped when no noise or vibration is heard. Check the
operator’s manual for additional procedures.
Avoid Being Caught in Moving Parts
Materials should be fed into the chipper/shredder safely, to avoid contact with the equipment’s
moving parts. Follow these feeding procedures to keep you safe.
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Feed material only when the chipper/shredder is at full operating speed.
Feed branches from the side of the chute, not in front of it. This will reduce the risk of
you being caught and dragged into the machine. Standing to the side of the equipment
will also make it easier for you to reach the emergency shut-off switch in the event of an
accident.
Keep hands and feet outside of the feed chute.
Use a push stick to help feed small pieces through the chipper/shredder to keep you at a
distance from the machine’s moving parts. Do not push materials into the chute with
your hands or feet, pitch forks, shovels, etc.
Let go of material as soon as it begins to be pulled into the machine and walk away to
avoid being hit or dragged into the chipper by limbs you are feeding.
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Feed the branches butt end first to keep the chipper from being jammed and to reduce
the kickback of material.
Lay shorter pieces of material on top of longer pieces and feed through the machine.
Collect small materials such as leaves and twigs and put them with the chipped material
instead of feeding them through the chipper/shredder.
Safe Fueling Procedures
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Gasoline and other fuels are flammable. Following safe fueling procedures will help
reduce the risk of fuel ignition.
Always shut off the engine and wait at least 5 minutes for the engine to cool before
refueling.
Use only approved fuel containers and store in a well ventilated area, away from direct
sunlight.
Never smoke or have an open flame near fuel.
Touch the fuel nozzle to the chipper/shredder before opening the fuel cap to reduce the
chance that a static spark will ignite the fuel.
Keep the nozzle in contact with the filler neck while fueling.
Replace the cap as soon as you are finished.
Use a funnel or non-spill nozzle when fueling to reduce spillage.
Wipe up all spills immediately, before starting the engine.
Never clean your hands or tools with gasoline. Use a nonflammable solvent instead.
Transportation & Traffic
Never move, load or transport a chipper/shredder while the engine is running. First, shut off
the power supply, wait for all moving parts to come to a complete stop and then disconnect the
spark plug wire.
Standard Operating Procedure – Grinding Feed for Chemical Analyses using a Wiley Mill
Laboratory mills are devices used for grinding of plant tissue or other similar materials. Use of
laboratory mills may present a variety of potential safety hazards.
Mill Set-up
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Use the right tool for the job. Laboratory mills such as the Thomas Scientific Wiley Mill
are intended for grinding plant samples; they should not be used to grind soil or other
hard materials.
Before using, always inspect the laboratory mill for damage or disrepair.
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If a laboratory mill fails the pre-use inspection, notify your supervisor and remove the
mill from service by attaching a red tag that states “DO NOT USE.” Complete red tag
with appropriate information.
Make sure the mill is unplugged before making any adjustments or cleaning.
With the mill unplugged, remove the cover and inspect the feed chute, grinding blades,
screen and delivery chute.
Confirm that the proper screen and delivery chute are attached and change as
necessary.
If parts of the mill need cleaning, use a small brush, a vacuum, and/or compressed air
(See Safety Note #153). Ethanol may be used if necessary to prevent contamination of
samples.
Operating Tips
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Make sure the sample is dry before milling; samples containing excessive moisture or
oils will stick to the walls of the mill chamber.
Use in a well ventilated area. If samples are excessively dusty, use the mill under a
ventilated hood, or use a fan to exhaust the dust outside the work area.
In general, respiratory protection should not be necessary when using a laboratory mill.
However, a paper dust mask (not an N-95 respirator) may be used voluntarily for
nuisance dust. If samples are suspected of containing molds or other disease-causing
agents, contact ANR Environmental Health & Safety (http://ucanr.org/askehs) to
conduct an exposure assessment to determine the appropriate personnel protective
equipment.
Wear eye protection (safety glasses or goggles) when using a laboratory mill.
Do not put your hands or fingers inside the feed chute when feeding samples into the
mill. If necessary, use a feeder stick to push material into the grinding chamber.
If the feed chute becomes clogged or material becomes jammed, immediately shut off
the mill.
Cleaning and Shutdown
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Clean the mill as described above in Mill Set-up.
It is a good practice to unplug the mill when not in use.
Consult the operator’s manual for maintenance procedures such as lubrication or blade
replacement.
Standard Operating Procedure – Building Pens for housing sheep
Section 1 - Personal Protective Equipment
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Long of loose hair must be tied back or contained
Work gloves required
Protective clothing required
No jewelry, watches, ring, necklaces
No loose fitting clothing
Close-fitting clothes
Long pants and turdy, non-slip boots
Section 2 - Potential Hazards + Safety precautions
You can be cut with the panels or the wire used to tie the panels. Work gloves are required as
well as non-slip boots, long pants and long-sleeve shirts to avoid skin cuts.
Standard Operating Procedure – Safe Sheep Handling
Catching sheep
It is easier to catch a sheep if it is part of a group, rather than by itself. Catching sheep is also
easier in a pen. The smaller the pen, the better. If you can, use gates or hurdles to make the
pen as small as possible. If you are in the paddock, bunch the sheep into a corner for catching.
Get the sheep into a corner, extending your arms to make a visual barrier. Approach the sheep
slowly and calmly. The sheep may try to escape but will probably not move away from the wall
or fence, so its moves can be anticipated.
If you can, come up quickly through the ‘blind spot’ directly behind the sheep. Grab it under the
chin. Do not grab the wool as this causes pain, distress and bruising.
Do not chase sheep around the pen or paddock. It’s tiring and potentially dangerous for both
the sheep and the catcher. Sheep that are repeatedly chased become flighty, stressed and
difficult to work with.
You can use a neck or leg crook, which gives you a longer reach, for catching sheep. You can
also use it to catch a sheep by the hind leg, but sometimes this can damage the legs and udder.
Controlled holding and walking
The easiest way to keep a sheep still is to stand it against rails or a fence and hold it with your
knees and a hand under the chin.
To walk a sheep, stand over the sheep’s shoulders with a leg either side of the sheep and your
hand under the chin. You can control the sheep with pressure from your knees. Walk the sheep
forward by squeezing it with your knees, and/or squeezing the top of the tail with one hand,
keeping your other hand under the sheep’s chin. It can be difficult for a short person to walk a
tall sheep in this way, so make sure it is within your capabilities.
Sitting a sheep up
The best way to sit a sheep up is to turn the sheep’s head on to its shoulder.
In a clear area, hold the sheep against your braced knees with one hand under its chin and one
on its rump. Turn the sheep’s head to the rear with one hand while forcing the rump down
against your leg with the other hand. When the sheep is no longer standing, lift the front leg
and sit the sheep securely on its rump.
Manually handling sheep
Manually handling sheep is a physically demanding job. Due to the size and strength of sheep,
you face problems like back strain and knee injuries when handling and restraining them.
Injuries also happen when sheep run into you and/or knock you over. Sheep are not usually
aggressive but, if they are frightened, they can hurt people when trying to get away. For
example, they may jump when stressed. They can jump with enough force to break a handler’s
leg, or high enough to strike a handler in the chest or face and knock them over.
Lifting sheep
Sheep are large and heavy animals and can fight against being lifted. This creates the potential
for strains and back twisting injuries.
Avoid lifting sheep if possible. Use gates and ramps where available. If you have to lift a sheep,
use your legs, not your back.
Only physically fit and strong people should lift sheep. No one should lift a sheep if it is too
heavy for them.
Standard Operating Procedures – Lewiston Irrigated
Pasture Facility
Standard Operating Procedure – Cattle Managing Procedures
Section 1 - Personal Protective Equipment
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Long of loose hair must be tied back or contained
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Work gloves required
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Protective clothing required
•
No jewelry, watches, ring, necklaces
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No loose fitting clothing
•
Close-fitting clothes
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Long pants and turdy, non-slip boots
Loading, unloading and transportation- Potential Hazards + Safety precautions
• Cattle sorting and holding pens should allow handling without undue stress, and be located
near the loading/unloading facility.
• All vehicles used to transport cattle should provide for the safety of personnel and cattle
during loading, transporting and unloading.
• Strictly adhere to safe load levels with regard to animal weight and space allocation.
• Personnel hauling cattle in farm and ranch trailers must ensure that adequate space is
provided so that cattle have sufficient room to stand with little risk of being forced down
because of overcrowding.
• When the vehicle is not full, safely partition cattle into smaller areas to provide load stability
for the cattle and the vehicle.
• No gap which would allow injury to an animal should exist between the ramp, its sides,
flooring, and the vehicle.
• Vehicle doors and internal gates should be sufficiently wide to permit cattle to pass through
easily without bruising or injury.
• Cattle should be loaded, unloaded, and moved through facilities with patience and as quietly
as possible to reduce stress and injury.
Animal Weighing- Potential Hazards + Safety precautions
• Measurement of live weight gain requires the fasting of animals to minimize the
variation caused by ‘gut-fill’ from digesta.
• Any cattle fasting for 12-24 hours must have water available in the holding pen.
• Weights of cattle are obtained on an individual basis to determine their daily gain. One
other purpose for weighing cattle individually is to use the weights to block the cattle
into weight groups or to sort animals into treatment groups.
• On weigh days, cattle are brought to the group pen by the scale located at the animal
working facility. The scale is zeroed, and it is checked periodically as animals within a
group are weighed to ensure that the indicator returns to zero when no animal is on the
scale. After all animals from a group have been weighed, the scale is checked for zero,
cattle are returned to their respective pastures, and the next group is weighed.
• Cattle should be handled quietly before, during and after the procedure.
• Adjust hydraulic chute to the appropriate size of cattle to be handled. Regular cleaning
and maintenance of working parts is imperative to ensure the system functions properly
and is safe for the cattle and handlers.
• Floors should be properly drained and barns and handling alleys should provide
adequate traction to prevent injuries to animals and handlers.
• Handling alleys and housing pens should be free of sharp edges and protrusions to
prevent injury to animals and handlers.
• Design and operate alleys and gates to avoid impeding cattle movement.
• When operating gates and catches, reduce excessive noise, which may cause distress to
the animals.
• Avoid slippery surfaces, especially where cattle enter a single file alley leading to a chute
or where they exit the chute. Grooved concrete, metal grating (not sharp), rubber mats
or deep sand can be used to minimize slipping and falling.
• Quiet handling is essential to minimize slipping.
• Under most conditions, no more than 2% of the animals should fall outside the chute.
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Take advantage of cattle’s flight zone and point of balance to move them. For safety and
welfare reasons, minimize the use of electric prods. Non-electric driving aids, such as
plastic paddles, sorting sticks, flags or streamers (affixed to long handles) should be used
to quietly guide and turn animals. When cattle continuously balk, cattle handlers should
investigate and correct the reason rather than resort to overuse of electric prods.
Tips for better cattle handling
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Cattle have wide angle vision in excess of 300°.
Position yourself adjacent to the head of the lead animal and at about 90° to the
direction the group is to be moved.
Be at a distance appropriate for the particular group (i.e., where your presence is
recognized but the lead animal does not attempt to move away from you).
Be on the side of the lead animal that you wish the group to turn away from (cattle are
uncomfortable losing eye contact with you and are reluctant to turn with their back
towards you).
Moving forward 90° will cause the lead animal to slow down, and moving behind 90° will
either speed the animal or encourage it to turn towards you.
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