Thermo Fisher Scientific FAMOS Well Plate Microautosampler User's manual

FAMOS

Well Plate Microautosampler
User’s Manual
P/N 160557
www.lcpackings.com
D931R1
Notice: The FAMOS Well Plate Microautosampler is covered by a limited
warranty. A copy of this warranty is included with this manual. The customer is
required to perform routine maintenance as described in the User’s Manual on a
periodic basis to keep the warranty in effect.
All information in this manual is subject to change without notice and does not
represent a commitment on the part of LC Packings, BV.
The material included in this manual is provided to assist users in the operation,
maintenance and repair of the FAMOS Well Plate Microautosampler. It is
assumed that the individual using this manual has sufficient training in the use of
analytical instrumentation and is aware of the potential hazards including (but not
limited to) electrical hazards, chemical solvent hazards and the exposure to
pressurized solvents.
© November 2001, LC Packings, BV - A Dionex Company. All rights reserved. No
part of this manual may be reproduced or transmitted in any form or by any
means without the written permission of LC Packings, BV.
The following are registered trademarks of LC Packings - A Dionex Company:
UltiMate, UltiChrom, FAMOS, Thermos, Switchos, Acurate, UZ-View, UVBooster, FluoBoost, Pepmap, µ-Dumper, µ-Fluidics, Fusica, Nano Series, Pico
Series, µ-Guard, µ-Precolumn, NanoPrecolumn.
Cheminert is a Registered Trademark of Valco Instruments Co, Inc.
Windows is a Registered Trademark of Microsoft Corporation.
Printed in the Netherlands.
D931R1
Table of Contents
WARRANTY
v
INSTRUCTIONS FOR RETURNING INSTRUMENTS
vii
WARNINGS AND SAFETY PRECAUTIONS
ix
CE DECLARATION OF CONFORMITY
xi
CHAPTER 1
System Overview
1.1
1.2
1.3
1.4
CHAPTER 2
Features and Design of the FAMOS Well Plate
Microautosampler
Principle of Operation of the Microautosampler
General Design of the FAMOS Well Plate Microautosampler
Contents of this Manual
2-1
2.1
2-1
2.2
2.3
2.4
Installation
Unpacking
The Standard Microautosampler Configuration
Electrical Connections
2.4.1
2.4.2
2.4.3
2.4.4
2.4.5
2.4.6
2.5
2.6
Inputs and Outputs
P4 Connector – MARKERS
P5 Connector - AUXILIARIES
P1, P2, P3 and P6 Connectors
Connector Communication
Power Connector
Connecting the Autosampler to the HPLC System
2.5.1
2.5.2
2.5.3
2.5.4
2.5.5
Preliminary Operations
Connecting Waste Tubing
Filling the Wash Solvent Bottle
Flushing the Syringe
Adjusting the Needle Height
Routine Operation of the System
2.6.1 Sample and Mobile Phase Considerations
2.6.2 Plates and Sample Handling
2.6.3 Placing Reagent Vials/Transport Vials in Position
User’s Manual FAMOS
1-1
1-2
1-3
1-4
Installation and Getting Started
2.1.1 Location of the FAMOS Well Plate Microautosampler in the
Laboratory
CHAPTER 3
1-1
2-1
2-2
2-3
2-5
2-5
2-6
2-6
2-6
2-6
2-7
2-8
2-8
2-9
2-10
2-10
2-11
2-12
2-12
2-12
2-13
The User Interface
3-1
3.1
3.2
3.3
3.4
3.5
3.6
3.7
3-1
3-2
3-3
3-5
3-6
3-7
3-8
Overview
Powering up the FAMOS Well Plate Microautosampler
The (Upper) Front Panel
Menus of the FAMOS Well Plate Microautosampler
General Approach to Entering and Executing a Program
Types of Methods
Executing a Series
D931R1
i
Table of Contents
Series Menu
3-8
3-9
3-10
3-10
3-13
3-13
3-15
3-16
3-16
3-17
3-17
3-17
3-18
3-19
3-20
3-21
3-22
3-25
3-26
Programming Examples
4-1
4.1
4.2
4.3
4.4
4-1
4-2
4-3
4-4
4-4
4-4
3.7.1 Manually Executing a Series
3.7.2 Executing a Series via Remote Control
3.8
Menus and Operating Commands
3.8.1 Ready Menu
3.8.2 The System Menu
3.8.3 The System Menu Item <GENERAL>
3.8.4 The System Menu Item <USAGE>
3.8.5 The System Menu Item <PLATES>
3.8.6 The System Menu Item <IO>
3.8.7 The System Menu Item <CLOCK>
3.8.8 The System Menu Item <COMM>
3.8.9 Methods Menu
3.8.10 The Method Menu Item <INJECTION>
3.8.11 The Method Menu Item <WASH>
3.8.12 The Method Menu Item <TIMEBASE>
3.8.13 The Method Menu Item <MIX>
3.8.14 The Method Menu Item <USER PROGRAM>
3.8.15 The Method Menu Item <TEMPLATE>
3.9
CHAPTER 4
Overview
General System Settings
Installing the Plate
Sample Applications
4.4.1 Conventions
4.4.2 Example 1: Injecting a Single 200 nL Sample from 1 vial
4.4.3 Example 2: Injecting a Single 100 nL sample from 3 vials and
Injecting 5 µL sample from 3 other vials
4.4.4 Example 3: 1µL injection out of a 5 µL sample in vial A1 using
the transport liquid (µL pick-up)
CHAPTER 5
5-1
5.1
5.2
5.3
5-1
5-2
5-3
5-3
5-3
5-5
5-5
5-5
5-6
5.4
Overview
Establishing a Test Protocol
The Test Protocol
Checking System Components
5.4.1 Testing the Accuracy of Delivery of the Syringe
5.4.2 Testing the Loop Volume
5.4.3 Testing the Tray Cooling Option
Maintenance and Troubleshooting
6-1
6.1
6.2
6.3
6-1
6-2
6-3
6-3
6-4
6-4
6-6
6-7
Overview
Maintenance
Replacing Major Components
6.3.1
6.3.2
6.3.3
6.3.4
6.3.5
ii
4-7
Testing the Microautosampler
5.3.1 The Analytical System used for the Test Protocol
5.3.2 The Test Protocol
CHAPTER 6
4-5
Replacing the Syringe
Replacing the Syringe Tip
Needle Assembly
Prepuncturing Needle
Combination Syringe, Sample Loop and Buffer Tubing
D931R1
User’s Manual FAMOS
Table of Contents
6.3.6 Replacing the Main Fuses and Setting the Operation Voltage
6.4
6.5
6.6
Troubleshooting
Error Codes of the Microautosampler
Spare Parts List
6.6.1
6.6.2
6.6.3
6.6.4
6.6.5
CHAPTER 7
Specifications
7-1
7.1
7-1
7-1
7-2
7-2
7-2
7-2
7-3
7-3
7-4
General
7.1.1
7.1.2
7.1.3
7.1.4
7.1.5
7.1.6
7.1.7
7.1.8
APPENDIX A
Major Items
Needle Assembly
Syringe and Buffer Tubing
Injection Valve
Vials and Accessories
Analytical Performance
Programming
Inputs/Outputs
Physical
Electrical
Communication
Options
Plate Dimensions
Injection Principles
A-1
A.1
A-1
A-1
A-3
A-4
A-4
A-5
A-6
A-6
A-8
A-9
A-10
Full Loop Injections
A.1.1 Standard Mode
A.1.2 Using an Air Segment
A.2
Partial Loopfill Injections
A.2.1 Standard Mode
A.2.2 Using an Air Segment
A.3
µL Pick-Up Injections
A.3.1 Standard Mode
A.3.2 Using an Air Segment
A.4
A.5
APPENDIX B
APPENDIX C
B-1
B.1
B-1
Overview of the Operating Program
Additional Programming Examples
C.4
C.5
C.6
C.7
User’s Manual FAMOS
Low Dispersion Injection
Comparing the Various Sample Injection Modes
Programming Charts
C.1
C.2
C.3
APPENDIX D
6-8
6-10
6-11
6-13
6-13
6-13
6-14
6-15
6-15
C-1
C-1
Overview
C-2
A 1.0 µL Partial Loopfill
A 3 x 1.0 µL Injection with µL Pick-Up and Wash between
C-4
Injections
A 1:10 Dilution followed by a 1.0 µL Partial Loopfill Injection C-6
Defining a Template and Adding a Protection Code
C-9
C-10
1µL injections out of 25 wells of a 384 well plate
C-12
Column Switching
The Sample Injector
D-1
D.1
D.2
D.3
D-1
D-2
D-2
Removing the Valve from the Microautosampler
Maintenance
Disassembly/Reassembly of the Valve
D931R1
iii
Table of Contents
D-2
D-3
D.3.1 Disassembly of the Valve
D.3.2 Reassembly of the Valve
APPENDIX E
Digital Inputs and Outputs
E-1
E.1
E.2
E-1
E-2
E-2
E-2
E-3
E-4
E-4
E-4
E-5
E-5
Introduction
Contact Closure Outputs
E.2.1 P1 Connector – OUTPUTS
E.2.2 P4 Connector – MARKERS
E.2.3 P5 Connector – AUXILIARIES
E.3
TTL Outputs
E.3.1 P2 Connector - TTL OUTPUTS
E.3.2 P3 Connector - TIMED OUTPUTS
E.4
TTL Inputs
E.4.1 P6 Connector - TTL Inputs
APPENDIX F
Programming Forms
F-1
APPENDIX G
Connecting the NANO Column (or other Fused Silica
Capillary)
G-1
G.1
G.2
G.3
G.4
G.5
G-1
G-1
G-2
G-3
G-4
Introduction
Using PEEK Fingertights
Fitting Assembly
Using Long PEEK Hex Style Nuts
Spare Parts Lists
I-1
INDEX
iv
D931R1
User’s Manual FAMOS
Warranty
LC Packings (Netherlands) BV, warrants that the products manufactured and sold
by it to be free from defects in material and workmanship for normal use and
service from the date of delivery to original purchaser for a period of one (1) year
from the date of shipment. This limited warranty does not cover, and no
warranty is provided, for parts that by their nature are required to be replaced
periodically as a function of use of the normal operation of the system. These
items include, without limitation: HPLC columns, fuses, tubing, detector sources,
pump piston seals, injector rotors, check valves, filters, any software, etc. In
addition, damage due to corrosion, misuse, negligence, accident, alteration of the
system or repair by an unauthorized individual is not covered by the warranty. It
is understood that the performance characteristics of the instrument require that
the mobile phase be degassed with He as described in the User’s Manual.
This warranty covers products sold under the LC Products trademark. If a
different warranty than the above is indicated in the sales literature, the warranty
indicated in the sales literature will prevail. If the system includes equipment
supplied by LC Packings but manufactured by a third party, LC Packings makes
no warranty of any kind, express or implied, including, without limitation, any
warranty of merchantability or fitness for a particular purpose. LC Packings will
make available to you, to the extent permitted, the warranties of the
manufacturer of the relevant equipment following your timely written request.
If any product covered by this warranty becomes defective during the warranty
period, it will be repaired or replaced by LC Packings at no charge to the
customer (the repair/replace decision is solely at the option of LC Packings). All
warranty requests must be received by LC Packings during the warranty period.
LC Packings will pay for surface transportation to the applicable LC Packings
Office (North America - San Francisco CA, Europe and Asia - Amsterdam, the
Netherlands), if the instrument proves defective within thirty (30) days from the
date of shipment (this does not include air freight, drayage, labor, crating
charges, customs clearance charges, etc.). The user should carefully follow the
directions indicated on the Return Goods Instruction Sheet in the User’s Manual.
After thirty days, all transportation costs will be at the expense of the customer.
Software Warranty
If, at any time during the period ending ninety (90) days after delivery of any
product to you, you report and document any error in any software provided
with such product and developed by LC Packings or any failure of any such
software substantially to conform to LC Packings software description that limits
or prevents use of the software by you, we will use reasonable efforts to correct
any such error or failure, will replace such software or will terminate your license
to use the software and refund the price of the related product. In connection
with any such termination and refund, you will return the related product to LC
Packings upon request.
The warranty will apply only to those portions of the software that were
developed by LC Packings and that incorporated all program corrections and
modifications, if any, delivered to you. It will not apply to any error or failure due
to machine error or to the misuse by or negligence of any person or entity other
than LC Packings or to any software, which is modified by any person, or entity
other than LC Packings.
User’s Manual FAMOS
D931R1
v
Warranty
Liability
Under no circumstances shall LC Packings be liable for damage to persons or
property. This warranty is the only warranty given by LC Packings with respect
to products and software provided with the products and is given in lieu of all
other warranties, express or implied, including, without limitation, any warranty
of merchantability or fitness for a particular purpose.
Your exclusive remedies and LC Packings’s sole liability for any non-conformity
or defect in the products and such software will be those expressed herein.
Under no circumstances will LC Packings’s liability arising from the performance
or failure to perform of any product or software, in contract, in tort (including
negligence), or otherwise, exceed the purchase price of the product and
software. In no event will LC Packings be liable, in contract, in tort (including
negligence), or otherwise for special, incidental, consequential or analogous
damages, including, without limitation, damages resulting from loss of use, loss
of profits, loss of business or loss of goodwill, even if LC Packings has been
advised of the possibility of such damages.
This warranty comprises the entire warranty between LC Packings and the
customer. It overrides any warranty related language that may appear in the
customer purchase order or other documentation provided by the customer.
This warranty shall be governed by, and construed and enforced in accordance
with, the laws of the Netherlands. It is non-transferable and shall run to the
benefit of the original purchaser only. Any change, alteration or amendment to
this warranty is not valid unless it has been approved in writing by an officer of
LC Packings.
vi
Warrantor North America
LC Packings (U.S.A.) Inc.
80 Carolina Street
San Francisco CA 94103
USA
Warrantor Europe and Asia
LC Packings (Netherlands) BV
Abberdaan 114
1046 AA Amsterdam
The Netherlands
Phone: (415) 552-1855
Fax:
(415) 552-1858
Phone: + 31 20 683 9768
Fax:
+ 31 20 685 3452
D931R1
User’s Manual FAMOS
Instructions for Returning Instruments
Before you return any item for repair, please contact the nearest LC Packings
office or its local distributor for instructions and obtain a return authorization
number.
Pack the equipment carefully, preferably in its original carton and ship it to the
LC Packings Service Department, using the appropriate address.
North America
LC Packings (U.S.A.) Inc.
80 Carolina Street
San Francisco CA 94103
USA
Europe and Asia
LC Packings (Netherlands) BV
Abberdaan 114
1046 AA Amsterdam
The Netherlands
Phone: (415) 552-1855
Fax:
(415) 552-1858
Phone: + 31 20 683 9768
Fax:
+ 31 20 685 3452
IMPORTANT:
1) Make certain that the return authorization number is indicated on the address
label of the package so that we can properly track and account for your
system.
2) Please include the following
a) Company letterhead with the following information.
•
•
•
•
•
•
•
Your Name
Complete Mailing Address
Telephone Number, fax number and e-mail address
Return Authorization Number
A detailed description of the problem.
The name of the LC Packings personnel to whom you have
spoken to regarding the problem
Return Shipping Information (if appropriate)
b) Relevant chromatograms
c) A purchase order (if the system is not in warranty)
User’s Manual FAMOS
D931R1
vii
Instructions for Returning Instruments
[This page intentionally left blank]
viii
D931R1
User’s Manual FAMOS
Warnings
The Danger sign, Warning sign and the Hazard sign shown below are included in
various locations in this manual. These signs provide the following information:
DANGER
Danger: The information in a danger statement relates to a procedure, practice
condition or action that if not done correctly or adhered to could lead to personal
injury or loss of life.
WARNING
Warning: The information in a warning statement relates to a procedure, practice
condition or action that if not done correctly or adhered to could lead to severe
injury and/or damage or destruction to parts or all of the equipment.
CAUTION
Caution: The information in a caution statement relates to a condition that could
lead to damage to equipment and/or lead to invalid analytical results.
Note: The information in a note statement relates to important information that
should be read and understood before continuing.
Safety Precautions
Note: The following precautions should be followed to minimize the possibility of
personal injury and/or damage to property.
Note: Make certain that you are familiar with the contents of this manual before
working on the system.
1) The system should be installed in a well-ventilated laboratory. If the mobile
phase includes volatile or flammable solvents, make certain that they are not
allowed to enter the workspace.
2) If the mobile phase includes volatile or flammable solvents, avoid open
flames and sparks.
3) If a leak occurs, turn off power to the instrument and remedy the situation
immediately.
4) All components of the system should be plugged into a common power line
that is directly connected to a true ground.
5) When the panels are removed dangerous electrical connections will be
exposed. Disconnect the autosampler from all power sources before
removing the panels.
6) Always replace blown fuses with fuses of the same size and rating indicated
on the fuse holder and panel. Refer to Section 6.3.6 of this manual for more
information on Fuses
User’s Manual FAMOS
D931R1
ix
Warnings and Safety Precautions
7) Repair or replace faulty power cords and all communication cables.
8) Many organic solvents and buffers are toxic. Make certain that you know the
toxicological properties of all mobile phases that you are using.
9) The toxicological properties of many samples may not be well known. If you
have any doubt about a sample, treat it as if it contained a potentially
harmful substance.
10) Wear protective eye goggles when handling mobile phases or operating the
instrument. An eye wash facility and a sink should be close to the unit. If any
mobile phase splash on the eyes or skin, wash the affected area and seek
medical attention.
11) Dispose of all waste mobile phase in an environmentally safe manner that is
consistent with all local regulations. Do not allow flammable and/or toxic
solvents to accumulate. Follow a regulated, approved waste disposal
program. Never dispose of flammable and/or toxic solvents through the
municipal sewage system
12) PEEK tubing is used in a variety of locations. While this polymer has superb
chemical resistance to most organic solvents, it tends to swell when it is
contact with CHCl3, DMSO and THF. In addition, it is attacked by
concentrated acids such as Sulfuric Acid and Nitric Acid (swelling or attack
by acid is not a problem with short flushing procedures).
Do not use PEEK tubing that is stressed, bent or has a kink.
13) Wear protective eye goggles when handling fused silica tubing (i.e.
installation, cutting etc.)
14) If a buffer is used as a part of the mobile phase, flush the system with
several volumes of a methanol/water (50/50) before it is shut down. This will
prevent salt buildup inside the unit.
15) Do not use the FAMOS Well Plate Microautosampler in ways other than
those indicated in the instructions given in this manual.
16) The following symbols are used on the FAMOS Well Plate Microautosampler:
W A R N IN G
S H A R P N E E D LE
A N D M O V IN G P A R T S
This indicates that care should be taken to prevent
personal injury or damage to parts of the FAMOS
Well Plate.
KEEP H AND S C LEAR
This sticker (with yellow background color) at the
back of the FAMOS Well Plate Microautosampler
calls attention to the fact that you are expected to
consult this manual for instructions on how to
operate the FAMOS Well Plate Microautosampler
x
D931R1
User’s Manual FAMOS
DECLARATION OF CONFORMITY
LC Packings Nederland BV
A Dionex Company
Abberdaan 114
1046 AA Amsterdam
The Netherlands
We
declare that our product
 Well Plate Microautosampler
FAMOS
is in confirmation with the following documents:
EEC directives 89/392, incl. 91/368 and 93/44 (machine safety) and EEC directives
73/23 and 93/68 (low voltage safety), applied with the following standard:
EN61010-1
Safety requirements for laboratory equipment
(Class I, Installation cat. II, Pollution degree II)
WARNING
LC Packings will not accept any liability for damages direct or indirect
caused by connecting this instrument to devices which do not meet relevant
safety standards.
EEC directives 89/336 and
standards:
EN 55011
EN 50082-1
EN 61000-3-2
92/31 (EMC requirements, applied with the following
Radio frequency emission
Voltage fluctuations
Harmonic current emissions
Use shielded cables and connectors for all remote connections.
Amsterdam, January 11, 2001
D932R1
User’s Manual FAMOS
Robert van Ling, QA manager
D931R1
xi
CE Declaration
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xii
D931R1
User’s Manual FAMOS
System Overview
CHAPTER 1
1.1 Features and Design of the FAMOS Well Plate
Microautosampler
The FAMOS Well Plate Microautosampler is a flexible and powerful system for
micro-HPLC and incorporates the following features:
• It can be used with a broad range of plates from various manufacturers
(96 well plate, 96 deep well plate, 384 well plate, 48 1.5 mL autosampler
vials, sealed or open).
• It is capable of executing full loop injections, partial loopfill injections and µL
pickup.
• It supports “Low Dispersion” injection control to optimize injection profiles.
• It can use a broad range of syringes (volumes of 25, 100, 250, 500 or
1000 µL).
• It is fully controlled by UltiChrom software.
• It is fully compatible with other laboratory equipment.
• It can be used for both routine analysis and method development purposes.
• It is menu driven and includes a broad variety of sample-handling programs
such as sampling, diluting, mixing and precolumn derivatization.
A number of options (e.g. Peltier plate cooling and a column oven) are available
for the FAMOS Well Plate Microautosampler. In addition, a broad range of
components to configure the unit to meet your requirements (e.g. loops, injection
needles, syringes, needle holder for nanospray) and disposable items (e.g. vials
and caps) are available. A detailed list of available items is presented in Section
6.6.
User’s Manual FAMOS
D931R1
1-1
System Overview
1.2 Principle of Operation of the Microautosampler
A schematic diagram of the FAMOS Well Plate Microautosampler is presented in
FIGURE 1-1.
Syringe Valve
Buffer tubing
Wash
Solvent
Injection
Valve
Needle pair
Syringe
FIGURE 1-1. Schematic Diagram of the FAMOS Well Plate Microautosampler
The FAMOS Well Plate Microautosampler includes the following components:
• Injection Valve - includes the loop to place the sample in the mobile phase.
• Needle Pair - used to pressurize the sample vial and withdraw sample.
• Buffer Tubing - provided to eliminate contamination of the syringe (especially
useful with small samples).
• Syringe - used to aspirate the sample from a well into the sample loop. To
prevent contamination of the syringe, the autosampler is equipped with buffer
tubing between the syringe and the injection valve.
• Wash solvent is used to remove the sample from the buffer tubing and sample
needle, and to rinse the buffer tubing and sample needle.
The FAMOS Well Plate Microautosampler provides three different methods of
injection for an analytical run:
• Full loop - The sample loop is completely (quantitatively) filled with sample.
This results in extremely good reproducibility.
• Partial loopfill - The sample loop is partially filled with sample. This provides
low sample loss. The operator can select the desired injection volume.
• µL pick-up - After aspiration of sample, the sample is transported into the loop
with transport liquid (mobile phase). In this mode, no sample is lost.
Technical information on the injection principle used by the FAMOS Well Plate
Microautosampler is presented in Appendix A.
1-2
D931R1
User’s Manual FAMOS
System Overview
1.3 General Design of the FAMOS Well Plate Microautosampler
The front view of the FAMOS Well Plate Microautosampler is presented in
FIGURE 1-2.
3a
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
Cover (in open position)
Buffer tubing
Injection valve including the Sample Loop (3a)
Tubing holder
Needle unit
Screw to fasten cover
Transport/reagent vials
Plate
Plate holder
Drain wash-position
Condensed water and leakage
Wash position
Wash solvent bottle
Syringe
Syringe waste tubing
FIGURE 1-2. Front View of the FAMOS Well Plate Microautosampler
User’s Manual FAMOS
D931R1
1-3
System Overview
1.4 Contents of this Manual
Note: This manual covers the standard version of the FAMOS Well Plate
Microautosampler as well as the inert version. If you are using an inert version,
please refer to Appendix G, which includes specific information that relates to
this configuration and the appropriate part numbers for replacement parts.
This manual describes the FAMOS Well Plate Microautosampler and includes the
following information:
Chapter 2:
Installation and Getting Started describes how to install the
FAMOS Well Plate Microautosampler.
Chapter 3:
The User Interface describes the use of the display panel and the
user interaction program to establish methods and series
Chapter 4:
Programming Examples presents details of a variety of
methods/series for that represent common applications of the microautosampler.
Chapter 5:
Testing the Microautosampler includes a protocol that can be
used to verify that the Microautosampler is operating in an acceptable manner.
Chapter 6:
Maintenance and Troubleshooting describes a variety of
maintenance procedures to optimize the performance of the microautosampler. In
addition, it discusses how the operator can determine the cause of a difficulty in
the operation of the autosampler and includes a list of spare/replacement parts
Chapter 7:
Specifications presents the specifications of the FAMOS Well
Plate Microautosampler
In addition, a series of appendices are provided to supply information about the
injection principles, a detailed chart that presents the operation program,
additional programming examples, a discussion about the injection valve and the
various inputs and outputs on the rear panel.
If you are using the FAMOS Well Plate Microautosampler with the LC Packings
UltiMate and/or UltiChrom software, please refer to the documentation provided
with these products for supplemental information. If the microautosampler is
used with other systems, the manuals provided with these systems should be
consulted for interfacing requirements.
1-4
D931R1
User’s Manual FAMOS
Installation and Getting Started
CHAPTER 2
2.1
Installation
The instructions provided below are provided for installation of the FAMOS Well
Plate Microautosampler as a stand-alone component in an HPLC system. When
the FAMOS Well Plate Microautosampler is used in conjunction with a LC
Packings UltiMate system, please refer to the UltiMate User’s Manual for
additional information.
Once you have set up the microautosampler, refer to Section 2.6 for information
about routine operation of the system.
2.1.1 Location of the FAMOS Well Plate Microautosampler in the
Laboratory
The FAMOS Well Plate Microautosampler should be installed in a facility with the
following environmental conditions:
•
The temperature range should be maintained between 10 and 40oC. The
system should be installed in an area in which the temperature is fairly
constant (do not place the system near a window, an air conditioning duct or
a heating duct). The humidity should be maintained between 20 and 80 %
relative humidity.
•
If flammable or toxic solvents are to be used, a suitable ventilation system
should be provided.
•
The use of open flames in the laboratory should be prohibited.
•
Corrosive vapors or dust should not be present as these materials can
adversely affect the long-term performance of the system.
The microautosampler requires approximately 280 mm (11.2”) of linear bench
space. The lab bench should be capable of supporting 100 kg (225 lb.).
The power consumption of the FAMOS Well Plate Microautosampler is 250 VA.
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Installation and Getting Started
DANGER
Danger: The FAMOS Well Plate Microautosampler must be connected to a power
source that is connected to a true ground. In addition, all other components of
the system (e.g. the HPLC pump, the detector) should be connected to the same
ground.
CAUTION
Caution: Do not install the FAMOS Well Plate Microautosampler in areas subject
to shock, dust, or in direct sunlight. Do not place it near a source of heat (this is
especially important if the tray cooling option is installed.
2.2
Unpacking
When the FAMOS Well Plate Microautosampler is received, carefully unpack the
unit and verify receipt of all components according to the packing list (some
components include sub-packing lists). It is recommended that all packing
materials be saved in the event that it is necessary to return any item to the
factory.
Note: When lifting the FAMOS Well Plate Microautosampler from the shipping
container, make sure that the unit is kept upright. Lift the unit by placing your
hands under the microautosampler.
If there is external damage to the shipping box, the damage should be reported
to the shipping agent and LC Packings upon receipt of the goods. If internal
damage is observed or if any items are missing, this should be reported to the
shipping agent and to LC Packings as soon as it is observed.
Note: If there is any apparent damage to the instrument, the user should
investigate the nature of the damage before plugging the unit into the mains to
ensure that powering up of the instrument will not create a hazardous condition
or damage internal components. If the damage appears significant, call LC
Packings or its local representative before connecting the unit to the mains.
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2.3
The Standard Microautosampler Configuration
A number of components of the FAMOS Well Plate Microautosampler are
factory-installed as presented in TABLE 2-1. The standard tubing configuration is
presented in TABLE 2-2.
If desired, these components can be replaced to meet the specific needs of the
analysis; a complete list of accessories and consumables is presented in Section
6.6. Changing the components is described in Section 6.3. Refer to FIGURE 2-1
to identify the various components.
TABLE 2-1. Factory Installed Items
Item
Injection Loop
Syringe
Buffer Tubing
Sample Needle
Wash Solvent Bottle
Fuses
(in power switch)
Description
5 µL
25 µL
50 µL
Fused silica, 2.4 µL
100 mL
115 V (AC) ± 10%:
two 5 AT fuses (slow, ¼” x 1¼”, UL/CSA)
230 V (AC) ± 10%:
two 2.5 AT fuses (slow, 5 x 20 mm, IEC127)
(The fuses used are UL-listed and CSA-certified)
TABLE 2-2. Standard Tubing Configuration
Tubing
Standard sample needle
and tubing
Buffer tubing from high
pressure valve to syringe
valve
Tubing syringe valve to
wash solvent bottle
Tubing syringe valve to
waste
User’s Manual FAMOS
Material and Dimensions
Fused silica tubing; 300 mm x 0.280 mm O.D. x
0.100 mm I.D. (total volume 2.4 µL)
PTFE tubing; 255 mm x 1/16" O.D. x 0.5 mm I.D.
(volume 50 µL)
PTFE tubing; 300 mm x 1/16" O.D. x 1.0 mm I.D.
PTFE tubing; 400 mm x 1/8" O.D. x 1.6 mm I.D.
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Installation and Getting Started
3a
5a
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
Cover (in open position)
Buffer tubing
Injection valve including the Sample Loop (3a)
Tubing holder
Needle unit with Sensor (5a)
Screw to fasten cover
Transport/reagent vials
Plate
Plate holder
Drain wash-position
Condensed water and leakage
Wash position
Wash solvent bottle
Syringe
Syringe waste tubing
FIGURE 2-1. Front View of the FAMOS Well Plate Microautosampler
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Installation and Getting Started
2.4
Electrical Connections
2.4.1 Inputs and Outputs
The FAMOS Well Plate Microautosampler has six standard I/O connectors (P1 P6), five OUTPUT connectors and one INPUT connector. The Communication
connector is a standard RS-232 communication interface connector. The
different configurations of the I/O connectors are described in Appendix E.
All electrical connections are made on the rear panel of the autosampler (FIGURE
2-2).
1
9
2
3
8
4
5
6
7
1
2
3
4
5
6
7
8
9
Serial Communication Connectors (RS232)
CE-mark
I/O connectors P1 - P3
I/O connectors P4 - P6
Power Entry Module with Main Switch
Fuses and voltage selector
Type label
Fan (if plate cooling is installed)
Ventilation holes
FIGURE 2-2. Rear Panel of the FAMOS Well Plate Microautosampler
CAUTION
Caution: Avoid touching the electrical contacts on the terminal strips.
Electrostatic discharges could damage internal components.
CAUTION
Caution: The manufacturer will not accept any liability for damages directly or
indirectly caused by connecting the FAMOS Well Plate Microautosampler to
instruments which do not meet relevant safety standards.
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Installation and Getting Started
The electrical connections that are required depend on the nature of the
instrumentation and the desired application. In this section, we describe a
standard installation with other LC Packings instrumentation.
If the system is interfaced to equipment from other manufacturers, the user
should refer to Appendix E for a detailed discussion of the various inputs and
outputs that are provided.
2.4.2 P4 Connector – MARKERS
If the microautosampler is installed with an UltiMate system, the P4 connector
should be connected to the START IN input of the UV Detector on the rear panel
of the UltiMate system. To connect the P4 MARKERS outputs, use the Inject
Marker Cable.
2.4.3 P5 Connector - AUXILIARIES
If the microautosampler is installed with a Switchos II Advanced Micro Column
Switching Unit and the UltiMate system, the position of micro valves of the
Switchos II as well as the gradient start are controlled via the P5 connector.
To connect the P5 AUXILIARIES outputs to the Switchos II and the UltiMate
system, use the (special) interface cable (P/N 160171).
2.4.4 P1, P2, P3 and P6 Connectors
This connectors are not used during an standard installation with the UltiMate
system (refer to Appendix E for more details when the microautosampler is used
with other systems).
2.4.5 Communication Connector
The FAMOS is equipped with two 9 pin RS-232 serial interfaces. In the standard
setup only the male type connector (S2) is used for digital transfer between the
autosampler and the PC (item 1, FIGURE 2-2). Some communication parameters
can be changed via two DIP switches above the RS-232 connectors. These
switches should be set to the default settings presented in FIGURE 2-3 and
should not be changed.
Default Settings
FIGURE 2-3. Default DIP Switch Settings
Note: Older versions of the FAMOS Well Plate Microautosampler are equipped
with only one 25 pin connector instead of two 9 pin connectors. If the
autosampler includes the DIP switches, they should be set to the default settings
presented and not be changed.
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2.4.6 Power Connector
The FAMOS Well Plate Microautosampler is equipped with a power supply for
input voltages from 110 to 120 VAC and 220 to 240 VAC. Changing of the
setting is only required if the voltage indicated by the voltage selector (FIGURE 24) on the back panel does not match with the mains voltage (FIGURE 2-4
indicates the setting for 220-240 V operation). The voltage indication plate can
be removed with a small screwdriver.
Voltage
Indication
FIGURE 2-4. Power Entry Module
The power cord should be inserted in the socket directly below the Main Power
switch.
CAUTION
•
Check whether local voltage matches voltage indicated on back panel of the
FAMOS Well Plate Microautosampler.
•
Connect the power cord to the FAMOS Well Plate Microautosampler (item 5,
FIGURE 2-2).
•
Switch the FAMOS Well Plate on by using the switch at the back panel (item
5, FIGURE 2-2).
Caution: Make certain that the instrument is properly grounded to a true earth
ground. Connecting the instrument to an ungrounded power line can cause
injuries and/or damage the instrument.
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Installation and Getting Started
2.5
Connecting the Autosampler to the HPLC System
2.5.1 Preliminary Operations
To install the FAMOS Well Plate Microautosampler to an HPLC system:
•
Place the FAMOS Well Plate Microautosampler in its operating location,
preferably on the left-hand side of the HPLC system. Make sure the
ventilation holes are not obstructed. Allow the instrument to acclimatize for 1
hour.
•
Loosen the screw at the right-hand side of the cover (item 6, FIGURE 2-1)
and lift the cover so that you can perform the procedures described in this
chapter.
•
Install the Plate Holder (item 1, FIGURE 2-5) in the FAMOS Well Plate
Microautosampler. It should be placed on the cooling plate (item 3, FIGURE
2-5) underneath the wash solvent bottle, as far to the left and to the back as
possible as shown in FIGURE 2-5. The gear wheel (item 2, FIGURE 2-6) must
fit against the teeth of the plate holder (indicated in white in the center of
FIGURE 2-6).
`400
3
2
1
FIGURE 2-5. Installing the Plate Holder
FIGURE 2-6. Bottom View of Plate Holder
2-8
•
Connect the HPLC pump to port 1 of the VICI-Valco injection valve.
•
Connect HPLC column to port 6 of the VICI-Valco injection valve.
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Installation and Getting Started
Note: The instrument has been flushed with isopropanol before shipment from
the factory. Make sure that the mobile phase of your HPLC system is miscible
with isopropanol, or start up with an intermediate solvent as mobile phase
(disconnect the HPLC column when flushing).
Note: It is very important that the contents of the sample loop are injected in
back flush mode onto the column, therefore: do not exchange column and pump
connections at the injection valve.
•
Power up the system. The display will indicate that the self-test and
initialization have been executed. After completion of this procedure, the
screen shown in FIGURE 2-7 will be presented.
12:04 THURSDAY JANUARY 04-01
READY (2.02)
[MENU]
<EXCHANGE> <WASH> <SYR END> <UTILS>
FIGURE 2-7. The Ready Menu
2.5.2 Connecting Waste Tubing
To connect the three waste tubes:
•
Syringe waste - put the end of the syringe waste tube (item 15, FIGURE 2-1)
in a bottle placed next to the FAMOS Well Plate Microautosampler.
•
Drain wash-position - connect the hose to the drain wash connector of the
FAMOS Well Plate (item 10, FIGURE 2-1); place the other end of the hose in
a bottle placed on the floor. All liquid dispensed to waste at the back of the
plate is removed through this drain.
•
Condensed water and leakage - connect the hose to the drain port of the
FAMOS Well Plate (item 11, FIGURE 2-1). Place the other end of the hose in
a waste container on the floor. All leakage of solvents and condensed water
are drained through this hose (if Peltier cooling option is installed).
Note: Make sure that the flow path of the hoses is not obstructed in any way.
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2.5.3 Filling the Wash Solvent Bottle
The FAMOS Well Plate Microautosampler is shipped with a 100 mL wash solvent
bottle.
To install the wash solvent bottle:
a) Fill the wash solvent bottle with the appropriate wash solvent. Use of mobile
phase in case of isocratic separations and mobile phase A – without any salt
or modifiers added to it – in case of gradients is recommended. Before using
the wash solvent, degas it with Helium or an ultrasonic bath, or degas it on a
continuous basis.
b) Screw the bottle to the cap in the holder.
c) Place the holder in the microautosampler as indicated in FIGURE 2-8.
d) Put the wash solvent tube in the wash solvent.
e) Flush the syringe (Section 2.5.4).
FIGURE 2-8. Wash Solvent Bottle
If you use an application that requires more than 100 mL of wash solvent for a
complete run, use a 250 mL wash solvent bottle (P/N 162033) or install a longer
tube (with flanged end for valve fitting) and place a larger bottle next to the
FAMOS Well Plate Microautosampler. To fill the wash solvent tube, you may
have to repeat the flushing procedure a few times.
2.5.4 Flushing the Syringe
The FAMOS Well Plate Microautosampler is supplied with a 25 µL syringe. It is
also possible to use the microautosampler with a 100 µL, 500 µL, 250 µl or
1000 µL syringe. Refer to Section 6.3.1 for instructions how to replace the
syringe.
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To remove the air and flush the syringe:
a) Check the general settings in the System Menu (Section 3.8) and make sure
that the appropriate syringe and the corresponding buffer tubing is installed.
b) Select soft function key <WASH> in the Ready Menu to perform a standard
wash routine. All tubing connected to the syringe valve is filled and rinsed.
c) If any air remains in the syringe, perform an additional wash cycle and gently
tap the syringe as wash solvent is dispensed to waste.
Note: If there is still air in the syringe gently tap the syringe as wash solvent is
dispensed to waste during the wash cycle.
2.5.5 Adjusting the Needle Height
The sample needle height can be programmed within the general system settings
(used for injection methods only), the mixed methods and within user defined
programs (Chapter 3). This parameter is defined as the distance from the top of
the plate holder to the bottom of the sample needle when the sample needle is
moved all the way down.
This parameter can be adjusted to insert the sample needle as far as possible into
the sample vial to aspirate the whole sample out of the vial (FIGURE 2-9). The
default setting is 2 mm.
Prepuncturing
Needle
Sample Needle
16 mm
Sample Vial
14 mm
10 mm
8 mm
Well Plate
6 mm
4 mm
Needle Height
12 mm
2 mm
Plate Holder
0 mm
FIGURE 2-9. Positioning the Sample Needle
CAUTION
Caution: A sample needle height ‘0’ corresponds to the top of the plate holder!
Operating the FAMOS Well Plate Microautosampler with this value set to ’0’ and
with a well plate installed at the same time may damage the sample needle. This
is especially true if the conventional stainless steel needle is used.
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Installation and Getting Started
2.6
Routine Operation of the System
2.6.1 Sample and Mobile Phase Considerations
The FAMOS Well Plate Microautosampler is used in an HPLC system and the
“standard” operating precautions for HPLC should be employed:
•
Ensure that samples and mobile phases do not contain particulate matter. All
samples and mobile phases should be filtered through a 0.22 µm membrane
filter. If organic solvents are used, make sure that extractable materials are
not present in the filter.
•
The sample should be soluble in the mobile phase. If a gradient is used, make
certain that the sample is soluble in the mobile phase at all mobile phase
compositions to be used in the separation.
•
After you have finished using the system, flush it with a water/methanol or
water/acetonitrile mobile phase before shutting it down.
•
Solvent should be degassed by sparging with He.
2.6.2 Plates and Sample Handling
The FAMOS Well Plate Microautosampler accommodates the following types or
plates:
•
96-low wells
•
96-high wells
•
384-low wells
•
48-vials
A sensor (item 5a, FIGURE 2-1) monitors plate detection, plate height detection
and vial detection.
Because the FAMOS Well Plate Microautosampler uses headspace pressure
during sample injections, it is very important that samples are properly handled.
Note the following:
•
Standard wells can best be filled by means of a narrow-end pipette to allow
air to escape when filling the well.
•
If wells are filled to the rim, the sample might be forced into the
prepuncturing needle, causing cross-contamination of samples and
contamination of the sample needle.
•
If vials are used (with a plate for vials), make sure the seals are airtight to
prevent air bubbles in the sample and prevent evaporation of volatile
samples; check seals after crimping; if the cap can be turned easily, the seal
is not airtight and the handcrimper should be adjusted.
Note: If wells that are not airtight are used, switch off the headspace pressure in
the System Menu (General Menu).
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Note: Check whether the needle height is sufficient for the new type of plate
that is installed and adjust if necessary (System Menu, General Menu).
To replace a plate in the FAMOS Well Plate Microautosampler:
a) Select the <PLATES> soft key in the Ready Menu, then select
<EXCHANGE>. The plate moves to the left.
b) Take out the plate and replace it by another one.
c) Select the <PLATE HOME> soft key. The plate moves to operating position
again.
d) If you have replaced the plate by a plate of the same type, you are now
ready. If you have installed a new type of plate, execute the following steps:
- Press System.
- Select the <PLATES> soft key.
- Press E.
- Select the soft function key for the type of plate concerned.
- Press E and determine whether to process the plate in <ROWS> or in
<COLUMNS>.
- Press Escape twice to return to the Ready Menu.
A message appears to indicate that all programmed series will be reset. The user
will have to redefine series because the settings in the System Menu have been
altered.
2.6.3 Placing Reagent Vials/Transport Vials in Position
To replace reagent vials/transport vials:
a) Select the <PLATES> soft key and then select <EXCHANGE> in the Ready
Menu. The plate holder moves to the left.
b) Take out the reagent vials/transport vials (item 7, FIGURE 2-1) and replace
them by other reagent vials/transport vials.
c) Select the <PLATE HOME> soft key. The plate moves to operating position
again.
e) Press Escape twice to return to the Ready Menu.
Note: Reagent and transport vials can be placed in any of the four positions.
Transport vials must be placed in a continuous row.
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The User Interface
CHAPTER 3
3.1
Overview
This chapter describes the general mode of operation of the FAMOS Well Plate
Microautosampler and explains how an operating program is established. It
includes a discussion on:
•
Powering Up the Microautosampler (Section 3.2)
•
A description of the front panel and the role of the keys on the front panel
(Section 3.3)
•
Menus of the FAMOS Well Plate Microautosampler (Section 3.4)
•
General approach to entering and executing a program (Section 3.5)
•
Types of Methods (Section 3.6)
•
Executing a Series (Section 3.7)
•
Menus and Operating Commands (Section 3.8)
•
Series Menus (Section 0)
Typical programs are presented in Chapter 4 and Appendix C.
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3.2
Powering up the FAMOS Well Plate Microautosampler
a) When the FAMOS Well Plate Microautosampler is powered up via the main
power switch on the rear panel, it will go through an initialization/self-test
protocol. During this period, a number of messages are displayed indicating
that various components are functioning properly.
b) After completion of this procedure, the Ready Menu appears on the display:
14:46
THURSDAY
JANUARY 04-01
READY (2.02)
[MENU]
<PLATES> <WASH> <SYR END> <UTILS>
FIGURE 3-1. The Ready Menu
Note: In this manual, the various display messages and menus shown correspond
to a system with firmware version V2.02. If a different version of the firmware is
used, there may be small differences in the screens and/or actions that occur
when a given command is performed.
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3.3
The (Upper) Front Panel
Communication between the user and the system is provided by the keypad on
the front panel of the FAMOS Well Plate Microautosampler (FIGURE 3-2) or via
the UltiChrom Software. This section describes the display and the use of the
keypad when the microautosampler is used on a local basis (i.e. not controlled by
UltiChrom Software).
Display
Soft Keys
Arrow Keys
Numeric Keys
Function Keys
FIGURE 3-2. Keypad and Display
The front panel includes:
•
Display - indicates a variety of system parameters.
•
Soft Keys - the label assigned to these keys depends on the menu that is
active. The current function of each key is shown in the bottom line of the
display.
•
Arrow keys - used to move to a different field in the display, to move to a
different field in a menu, or to change the value of a field.
•
Numeric Keys (0 to 9) - used to enter values in the various programming
fields.
•
CL(Clear) - used to clear a value in a field or replace it by NONE or AUTO.
•
E (Enter) - used to accept the present value or setting for an entry. The
entered value is checked for validity and then saved.
Function keys:
a) Run control keys:
•
Start/Stop - used to start or stop automatic processing (and data acquisition
when controlled via the output connectors) or to reset the system after an
error has occurred.
•
Hold/Cont.- used to hold or continue the analysis time. The analysis time is
extended by the period that Hold is active.
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•
Interrupt - not used
•
Priority - used to stop a run so that a priority sample is run before analyzing
the rest of the programmed sample series. Before the run is interrupted,
processing of the present sample will be finished. As soon as the priority
sample has been analyzed, the analytical run is resumed. A priority sample is
a series of one well with an injection method, a wash method and a time
base method defined in a template (this run is possible only if the correct
settings are entered in the System Menu).
b) Programming keys:
•
Series - used to enter the Series Menu in which series can be defined for an
analytical run.
•
Methods - used to enter the Methods Menu in which methods can be
programmed for use in an analytical run.
•
Menu - this key can only be used if [MENU] or [MN] is shown in the top
right hand corner of the display. If this key is pressed, additional fields of the
menu are displayed.
•
System - used to enter the System Menu in which system settings can be
entered.
c) General keys:
3-4
•
Escape- allows the user to leave the programming mode or go to a previous
level in the menu. A value that is entered is checked for validity and then
saved.
•
Help - used to display help information (which is available only for a limited
number of functions).
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3.4
Menus of the FAMOS Well Plate Microautosampler
The software of the FAMOS Well Plate Microautosampler is menu-driven. In this
section, we present an overview of the menu structure, as well as a detailed
description and a menu reference (name) for each command (Section 3.8). In
addition, a detailed programming flow chart is presented in Appendix B).
To access a menu, click on the soft key that corresponds to the menu. A few
menus have a sub menu associated with it; in these cases, the desired menu is
chosen in the standard way after the top level menu is accessed. When you are
in a menu, various items are accessed, and you should indicate the appropriate
choice(s) to generate the desired method or series.
The microautosampler menus are:
•
Ready Menu - this menu appears when the FAMOS Well Plate
Microautosampler has been powered up and passed the initialization test. It
includes basic system commands (e.g. washing and plate handling ), a
number of utilities (e.g. copying a method) and commands to establish
communication with other systems.
•
System Menu - this menu is used to select settings that are changed
infrequently (e.g. the loop volume, the type of plate, the system clock, if the
alarm should be set, etc.). In some cases, these settings may have been
made at the factory to suit particular wishes of the user; in this instance, it
will not be necessary to make changes. It is recommended that you enable
only those facilities that you will actually need; this will ensure that menus
are as concise as possible.
•
Methods Menu - this menu is used to program a method to be used in an
analytical run and to assign a number to it. Typical commands include the
analysis time, the wash volume and wash parameters.
•
Series Menu - this menu is used to define a series, and to assign a number
and a method to it for the analytical run.
If [MENU] or [MN] is displayed in the top right hand corner of the screen, you
can press the Menu key on the keypad to display more possibilities offered by
the menu. An explanation of all keys on the keypad is presented in Section 3.3.
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3.5
General Approach to Entering and Executing a Program
After you have determined the type of analytical run that you want to perform,
the best way to generate the order of operation for the FAMOS Well Plate
Microautosampler is:
a) Enter the desired settings in the System Menu. It is probable that the settings
in the System menu have been correctly entered (factory-installed) as these
settings are global in nature.
b) Program a method for the analyses you wish to perform via the Methods
Menu.
c) Define a series and link a programmed method to a range of wells in the
Series Menu.
d) Execute the series.
While you could use a different order than that indicated above, it should be
noted that some of the settings in the System Menu define the options in other
menus. If an invalid selection is made on the System Menu, the desired option
for other menus may not be presented. As an example of this point, the USER
PROGRAM menu (which is defined in Section 3.8.14) will be presented only if
the USER PROGRAM option has been selected in the System Menu (Section
3.8.2).
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3.6
Types of Methods
The FAMOS Well Plate Microautosampler provides the following types of
methods for different aspects of the operation of the microautosampler:
•
injection method - contains information on the injection routine, flush volume
and analysis time.
•
wash method - describes a wash volume and when a wash must be
executed.
•
mix method - a pre-injection method in which additional sample handling can
be performed (e.g. pre-column derivatization).
•
timebase method - a post-injection method with which outputs to other
devices (e.g. integrator or pump) and switching of the ISS valve are
controlled.
•
user program - offers the possibility to program sequences of all actions that
can be executed by the FAMOS Well Plate Microautosampler in separate
steps.
Each programmed method is assigned a number. The FAMOS Well Plate
Microautosampler allows you to store a combination of defined methods in a
template, which is also identified by a number.
Methods must be linked to a series before they can be used. The following
options are available with the FAMOS Well Plate Microautosampler:
•
You can assign an individual method to a series: methods (mix, injection,
wash, timebase) can be linked to wells in a series.
•
You can assign a template to a series: a combination of various programmed
methods (mix, injection, wash, timebase) can be defined in a template. The
template is linked to a range of wells in a series. In this way, all steps in an
analytical run are defined and stored.
•
You can assign a user program to a series: This can be used to combine all
possible steps in the analytical process in one program. The user determines
the order of the separate actions the FAMOS Well Plate Microautosampler
has to perform.
User’s Manual FAMOS
D931R1
3-7
The User Interface
3.7
Executing a Series
3.7.1 Manually Executing a Series
A series can be run from the Ready Menu by pressing the Start/Stop key.
Execution of a series is possible if you have programmed a method and defined a
series for the samples you wish to analyze.
Note: Series are not stored in battery backup memory and exist only for as long
as the FAMOS Well Plate Microautosampler is powered up.
To execute a series:
a) Start programming the series by pressing Start/Stop.
b) Enter the number of the first series to be performed and press E (to confirm
the input and go to the next screen), then indicate the number of the last
series to be performed and press E again.
c) After the range of series is defined, the autosampler will ask for the start
conditions. It is possible to start the series either from the keypad or by
remote control inputs (NEXT INJECTION and NEXT VIAL inputs, connector
P6).
d) Select <START> to manually start the analytical run. The FAMOS Well
Plate Microautosampler will begin execution of the series that you have
defined.
During the run, the display shows information about the current operation.
Note: Series are always executed in numerical order. Empty series will be
skipped.
You can stop a run by pressing the Start/Stop key. The FAMOS Well Plate
Microautosampler will execute a shut down sequence by removing all sample
from the buffer tubing and performing a wash routine.
If you want to stop the microautosampler immediately (panic stop), without
performing a buffer clean up; press the Start/Stop key twice. Remember to
perform a wash routine to clean up the tubing before you start the FAMOS Well
Plate Microautosampler again.
CAUTION
Caution: A panic stop does not remove the sample from the buffer tubing. Before
starting again, you should perform a wash sequence.
After the FAMOS Well Plate Microautosampler has completed the run, the Ready
Menu will appear again.
A number of examples of the use of a series are presented in Chapter 4 and
Appendix C.
It is possible to program a series and/or a method during a run by pressing Series
or Methods. The menus that are presented in this mode are identical to those
offered when the FAMOS Well Plate Microautosampler is idle.
3-8
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User’s Manual FAMOS
The User Interface
If a series or method is changed, the new values become active the next time the
FAMOS Well Plate Microautosampler initiates a series. The series currently
running are not affected by the changes.
3.7.2 Executing a Series via Remote Control
After the range of series has been programmed, the FAMOS Well Plate
Microautosampler will ask for the Start conditions. If you press the <REMOTE>
soft key, the Remote mode of operation will be selected. In this mode, the
autosampler will act as a slave of another device (e.g. an HPLC pump) and can
be controlled with the NEXT INJECTION and NEXT VIAL inputs of connector P6
(see Appendix E).
•
NEXT INJECTION INPUT - this input will start the next injection of the series.
If the series was completed, it starts the next injection from the next series,
until all series is completed.
•
NEXT WELL INPUT - this input will start the first injection from the next vial
in the series. If the series was completed, it starts the first injection from the
first vial of the next series, until all series are executed.
Note: The difference between the NEXT INJECTION and the NEXT WELL INPUT
is that the NEXT WELL INPUT directly starts the first injection from the next vial,
even if not all programmed injections from the previous vial were executed.
To indicate that the remote control mode is activated, the letter “r” is displayed
in the lower left corner of the display.
To execute a series via remote control, execute the following steps:
a) Press Start/Stop.
b) Enter the number of the first and the last series to be performed (each entry
should be confirmed by the E button).
c) Select <REMOTE> to enter the remote control mode. The FAMOS Well
Plate Microautosampler will now operate as slave of another device and can
be controlled with NEXT INJECTION INPUT or NEXT WELL INPUT.
d) Press Escape to return to the Ready Menu.
At the end of the series, the message “Series completed via remote control” is
displayed.
User’s Manual FAMOS
D931R1
3-9
The User Interface
3.8
Menus and Operating Commands
This section describes all the features and possibilities of the soft keys included
in the FAMOS Well Plate Microautosampler firmware, in the order in which they
appear on the screen. An overview of the programming scheme is presented in
Appendix B.
The different levels of the soft function keys (commands) on the different menus
are indicated as follows:
<PLATES>
(<bold with brackets>) represents the top level.
<EXCHANGE>
(<normal type with brackets>) represents second level.
NEEDLE HEIGHT
(normal type) represents the different parameter names.
3.8.1
Ready Menu
The Ready Menu contains the soft function keys presented in FIGURE 3-3.
Overview
<PLATES>
<WASH>
<SYR END>
<UTILS>
Second page of the [Menu]
<SSV>*
<SERIAL>
<COOL>*
<SERVICE>
*) Only if option is installed
FIGURE 3-3. The Ready Menu
All available soft function keys and options on the Ready Menu lead to the
following menu:
<PLATES>
<EXCHANGE>
<PLATE HOME>
<WASH>
<SYR END>
3-10
This key is used to access commands that are used to
exchange well plates.
Press <EXCHANGE> to move the plate to the left; in this
position the plate can be replaced without damage to the
equipment.
Press the soft function key <PLATE HOME> to move the
plate to operating position again.
This key is used to start a standard wash procedure. All tubing
connected to the syringe valve will be filled and rinsed with
wash solvent.
This key is used to move the syringe to the end position (e.g.
if you wish to replace the syringe needle or to simplify filling of
wash solvent tubing). A syringe volume of wash solvent is
aspirated from the wash solvent bottle and the wash solvent
tube is filled. When the key is selected, it is redefined to
<SYR HOME>, which is used to dispense the syringe
contents to syringe waste and to move the syringe to the
standard operating position again.
D931R1
User’s Manual FAMOS
The User Interface
<UTILS>
This key is used to enter the Utilities Menu.
Note: If a method protection code is enabled in the
System Menu, the code must be entered to access the
Utilities Menu.
<COPY>
This key is used to copy a method, select the method type
(mix, injection, timebase, wash), the number of the method to
be copied and enter the number for the destination method. An
existing method stored under that number will be overwritten.
<ERASE>
This key is used to erase a method (Template, Methods, User
Program). If Template and User Program are disabled in the
System Menu, the soft function keys for erasing a standard
Method (mix, injection, wash, timebase) appear. It is not
possible to erase the user program if the protection code for
the user program is enabled in the System Menu.
<LOG>
The FAMOS Well Plate Microautosampler keeps a log of
system-relevant events (<EVENTS>; records error messages
that have been generated) and keeps count of actions of
valves and syringe movements (<COUNT>). The message
“Lifetime of syringe (valve) may be exceeded. Check for
possible leakage!” appears after every 50,000 syringe actions
and after every 200,000 syringe valve actions.
Note: If you do not replace the Syringe when the
message is presented, and tell the system “not to
display this message again”, the message will not be
displayed again until an additional 50,000 more syringe
actions have been counted.
Note: If the Syringe valve is replaced, the counter
should be reset to zero by the service engineer.
<DEFAULTALL>
This key is used to change all software settings to default. All
series, methods, templates and the user program (unless
protected by protection code) will be erased.
Note: If <DEFAULT ALL> is selected, check whether
the hardware configuration is still compatible with the
settings entered in the System Menu.
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The User Interface
<SERIAL>
This key is used to set the FAMOS Well Plate
Microautosampler to serial mode.
Note: If a method protection code was defined in the
system settings, the code must be entered to get
access to serial mode.
<PANIC>
This key is used to initiate a stop sequence. A panic stop does
not remove the sample from the buffer tubing. Before starting
again, you should perform a wash sequence in which all tubing
is rinsed and the valve and I/O ports are reset. At the end of
the sequence, serial mode operation is resumed.
<EXIT>
Press to end serial mode and return to the Ready Menu.
<SSV>
(option)
This soft key is used to start a procedure in which all lines of
the solvent selection valve can be primed (if this option is
included in the system).
<COOL>
(option)
This soft key is used to enter the programming mode for
Peltier plate cooling (if this option is included in the system). If
the cool option is switched <ON>, the following soft function
keys can be selected:
<MANUAL>
The temperature control will remain OFF until it is switched on
again by the user (in this menu).
<AUTOMATIC>
<DATE-TIME>
The temperature control will be switched OFF after all
programmed series have been executed.
The temperature control will be switched OFF at a date and
time that can be user selected.
Note: The programmable temperature range is 4°°C to
40°°C. The maximum cooling capacity is approximately
20°°C below ambient. Make certain that the condensed
water and leakage tubes are connect to a waste
container on the floor to drain condensed water.
Note: The temperature of the sample inside the sample
vial may be slightly different than the temperature set,
due to a variety of effects (e.g. the heat transfer
characteristics of the vial walls). If it is necessary to
precisely set the temperature of the sample, we
recommend that you determine the temperature inside
the vial at various temperature setpoints and set the
cooling option so that the desired internal temperature
is attained.
<SERVICE>
3-12
This key is used to access a variety of service related
commands and it is protected by a code. Use of these
commands is restricted to authorized service personnel.
D931R1
User’s Manual FAMOS
The User Interface
3.8.2
The System Menu
When you press the System key, the display asks whether the autosampler
should operate in the <MICRO> or in the <CONVENTIONAL> mode.
<MICRO>
<CONVENTIONAL>
The ranges and the default settings of some system parameters are dependent
on this selection of the mode of operation.
Note: If you change from one operating mode to the other, all parameters will be
set to their default values for that mode.
In both operating modes the System Menu offers the soft function keys
presented in FIGURE 3-4 and which are discussed in detail in Sections 3.8.3 –
3.8.8 (<CLOCK> and <COMM> are available after pressing the Menu key).
<GENERAL>
<USAGE>
<PLATES>
<IO>
• Loop volume
• Needle tubing
• Syringe volume 1)
• Syringe speed
• Needle height
• Skip missing wells
• Air segment
• Headspace pressure
• Time base display
• Key click
• Error buzzer
• Alarm buzzer
• Method protection
code
• Use time based
method
• Use mix method
• Use user-program
• Use labeled wells
• Use templates
• Use calibration wells
• Type of plate
• Process in
ROWS/COLUMNS
• Number of transport
vial
• Inj. Marker pulse
• Well marker pulse
• Lab. Well marker
pulse
• Input edge next inj.
• Input edge next well
• Freeze input active
• Reset outputs after
series
.. <CLOCK>
• System clock
• Date
• Time
<COMM.>
• Device identifier
FIGURE 3-4 The System Menu
3.8.3
The System Menu Item <GENERAL>
Note: The microautosampler can be fitted with different syringes, tubing lengths,
etc. to meet the specific needs of the analysis. The General Menu is used to
enter the system configuration; it is critical that they are entered so that various
operations are performed properly (e.g. the minimum flush volume is dependent
on the needle volume). Once these settings are established, they are not changed
frequently.
<GENERAL>
LOOP VOLUME
NEEDLE TUBING
SYRINGE VOLUME
SYRINGE SPEED
SCALE FACTOR
…
User’s Manual FAMOS
This soft function key is used to access the following
parameters:
These values are entered by the user so that the system
can determine the appropriate values for certain
operations. As an example, the minimum flush volume is
dependent on the specific needle and needle tubing. The
default flush volume equals two times the volume of
needle and tubing.
The maximum aspiration speed of the syringe used in
injection methods is dependent on the viscosity of the
samples. As an alternative, the syringe speed can be
reduced by entering a scale factor. The actual syringe
speed will be the scale factor multiplied by the syringe
speed. The speed of the syringe during the wash or the
rinsing procedure of the buffer are not affected by this
setting.
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The User Interface
NEEDLE HEIGHT
This parameter refers to the distance between the needle
point and the plate holder (default: 2 mm). The value is
only used in injection methods (for mix methods this
value is programmable in the method itself).
SKIP MISSING VIALS
Appears only if a plate with 48 vials is selected in
System Menu (Plates Menu). YES means that empty
spaces are skipped during the run. NO means that the
Microautosampler will stop if an empty space is observed
during the run and an error code will be generated.
AIR SEGMENT
HEADSPACE
PRESSURE
This command is used to indicate whether an air
segment will be used for analytical runs (for explanation
of air segmentation refer to Appendix A).
This command is used to indicate if the headspace
pressure should be on or off. The FAMOS Well Plate
Microautosampler uses headspace pressure to facilitate
transport of sample into the loop. The compressor will
always be used during a wash procedure.
Note: Accuracy and reproducibility may decrease
if headspace pressure is switched off. However,
headspace pressure will only be useful if sample
wells are airtight
TIME DISPLAY
KEY CLICK
ERROR BEEP
ALARM BUZZER
This command is used to offer a choice between two
modes of representing the time.
These commands are used to allow the user to present a
audible sound when a key is pressed, when an error is
observed or if an alarm is observed.
TABLE 3-1 presents an overview of the default general settings and their ranges.
TABLE 3-1. Overview of General settings
<GENERAL>
Parameter
Loop volume
Needle tubing
Default Setting
Micro
Conventional
5 µL
100 µL
2.4 µL
15 µL
Syringe volume
25 µL
250 µL
low
0.1
normal
0.1
Syringe speed
Scale factor
Needle height
Skip missing vials
Air segment
Headspace
pressure
Time base display
Key click
Error beep
Alarm buzzer
Note:
3-14
2 mm
yes
yes
Range
Micro
Conventional
1 - 20 µL
5 - 1000 µL a)
1 – 99.9 µL
1 - 200 µL
fixed to
100, 250, 500
25 µL
or 1000 µL
low, normal or high
0.1 - 1.0
0 - 40 mm
yes or no
yes or no
no
yes or no
HH:MM:SS
on
on
on
H:MM:SS or H:MM:mm
on or off
on or off
on or off
a) 250/500 µL syringe: use 500 µL buffer tubing
500/1000 µL syringe: use 2000 µL buffer tubing
D931R1
User’s Manual FAMOS
The User Interface
3.8.4
The System Menu Item <USAGE>
<USAGE>
This soft function key is used to access the following
settings:
PROTECTION CODE
This entry is used to enter a code for protection of
methods. A six digit code (000000-999999) can be
selected and is erased by pressing CL. If a code has been
defined, it is not possible to enter the System Menu and
the programming menus without entering the protection
code. Default: none.
TIMEBASE
METHODS
This entry is used to enable/disable the ability to program
timebase methods. The microautosampler controls other
connected equipment during analysis time. Timebase
methods are programmed via the Methods Menu.
Default: disabled.
MIX METHODS
USER PROGRAM
This entry is used to enable/disable the ability to program
mix methods for the microautosampler. Program mix
methods in accessed via the Methods Menu.
Note: The microautosampler cannot analyze
priority samples during a run if the mix method is
enabled. Default: disabled.
This entry is used to enable/disable the possibility to
generate a user program. If this function is enabled, it is
possible to enter a user program protection code (6
digits). A user program is generated via the Methods
Menu. Default: disabled
Note: The microautosampler cannot analyze
priority samples during a run if the user program
is enabled.
LABELED WELLS
This entry is used to enable/disable the possibility to
program labeled wells. The location of labeled wells is
programmed via the Series Menu. Default: disabled
TEMPLATES
This entry is used to enable/disable the possibility to
program templates. Templates are programmed via the
Methods Menu. Default: disabled.
CALIBRATION
WELLS
This entry is used to enable/disable the possibility of
programming calibration wells. The location of the
calibration wells is entered in the Series Menu. Default:
disabled.
Note: We recommend that you disable as many
functions in the Usage Menu as possible to make
sure that other menus do not contain possibilities
that are irrelevant for the type of analyses you are
to perform.
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The User Interface
3.8.5
The System Menu Item <PLATES>
<PLATES>
TYPE OF PLATE
This soft function key is used to access the following
settings:
This entry is used to define the type of plates that will be
used. Four types can be selected: 96-low (default), 96high, 384-low or 48-vials. After a plate type has been
selected, press ENTER:
WELL PROCESSING
METHOD
Indicates if samples should be processed in rows (left to
right) or in columns (top to bottom).
FIRST TRANSPORT
VIAL
Enter a number 1 – 4, or press CL.
LAST TRANSPORT
VIAL:
Enter a number 1 – 4.
Note: Vials can be placed in any of the four
positions. Transport vials must be placed in a
continuous row.
3.8.6
The System Menu Item <IO>
<IO>
INJECT-MARKER
PULSE LENGTH
WELL-MARKER
PULSE LENGTH:
LABELED WELL
MARKER PULSE
LENGTH
INPUT EDGE NEXT
INJECTION
FREEZE INPUT
ACTIVE
RESET OUTPUTS
AFTER LAST SERIES
This soft function key is used to access the IO
configuration mode and define the following (the default
and range are indicated in TABLE 3-2):
This entry is used to define the length of the injectmarker pulse.
This entry is used to define the length of the well-marker
pulse.
This entry is used to define the length of the well-marker
pulse of the labeled well.
This entry is used to define the edge sensitive inputs for
the next injection.
This entry is used to define whether the freeze input is
active when high, or freeze input is active when low.
This entry is used to indicate whether the outputs should
be reset to default after the last series.
Note: Refer to Appendix E for more specific
information on IO connections.
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The User Interface
TABLE 3-2. Overview of IO settings
IO Parameter
Inject-marker pulse length
Well-marker pulse length
Labeled well marker pulse length
Input edge next injection
Input edge next well
Freeze input active
Reset outputs after last series
3.8.7
Range
0.1 - 2.0
0.1 - 2.0
0.1 - 2.0
falling or rising
falling or rising
low or high
yes or no
The System Menu Item <CLOCK>
<CLOCK>
3.8.8
Default
1.0 s
1.0 s
1.0 s
falling
falling
low
no
This entry is used to switch the system clock on or off. If
the ON selection is made, you can set date (yy:mm:dd)
and time (hh:mm). This date and time will be displayed in
the Ready Menu.
The System Menu Item <COMM>
<COMM.>
This entry to define a device identifier for communication
with other equipment (e.g. a PC). An identifier between
20 and 29 can be selected for the FAMOS Well Plate
Microautosampler, default setting is 21.
Note: When controlling the FAMOS Well Plate
Microautosampler by UltiChrom, make certain
that the same setting is used in the hardware
description.
3.8.9
Methods Menu
When entering the Methods Menu (by pressing the Method key), the user can
program various types of methods. It is possible to define up to:
•
24 Separate Injection methods
•
5 Wash methods
•
5 Timebase methods
•
9 Mix methods
•
1 User program
It is possible to program a combination of methods and save them in a template.
The available parameters and options in a method or template may vary
according the settings made in the System Menu.
An overview of the Methods Menu is presented in FIGURE 3-5.
User’s Manual FAMOS
D931R1
3-17
The User Interface
Methods
a)
<INJECTION>
<WASH>
Injection methods
<FULL>
•
•
•
Analysis time
Flush volume
Injections/well
•
Inject. control:
STD/L.D.
Flowrate
L.D. -factor
•
•
<PARTIAL>
c)
•
•
•
•
Analysis time
Flush volume
Injections/well
Inj. Volumes
•
Inject. control:
STD/L.D.
Flowrate
L.D. – factor
•
•
Notes:
<PICK-UP>
•
•
•
Analysis time
Injection/well
Inj. volumes
•
Wash volume
•
Wash
between:
– Series
– Wells
– Injections
•
•
•
•
•
•
•
•
<TIMEBASE> b)
<MIX> b)
Timebase program:
Mix method:
4x AUX 1
4x AUX 2
4x AUX 3
4x AUX 4
4x ISS-A
4 x ISS-B
8x CODE OUT
END TIME
c)
•
•
•
•
•
ASPIRATE
– sample
– destination
– air
– reagent
DISPENSE
– sample
– destination
– waste
– reagent
REPEAT
WAIT
WASH
a) <Template> and <User Program> disabled in System Menu
b) When enabled in System Menu only
c) In micro mode only
FIGURE 3-5. The Methods Menu
Note: The options provided by the Methods Menu depend on the settings made
on the System Menu.
3.8.10 The Method Menu Item <INJECTION>
<INJECTION>
<FULL LOOP>
<PARTIAL LOOPFILL>
<µL PICK-UP>
ANALYSIS TIME
FLUSH VOLUME
This soft function key is used to access commands to
program a method that defines injection methods for full
loop, partial Loop and µL pick-up operation. Up to 24
methods can be programmed.
Select the desired injection method and then enter a
number to define the injection method to be
programmed. If the selected method is locked because of
changes in the settings (System Menu) after
programming the method, the word LOCK is displayed.
The method can be unlocked by programming valid
values in the method itself or by restoring the values in
the System Menu.
This entry defines the time between switching the
injection valve to inject and the start of processing the
next sample.
This entry defines the amount of sample taken from a
vial before the loop is filled with sample. Default value:
5.0 µL (combined with an air segment).
Note: Flush volumes of less than twice the
volume of the needle and tubing will result in
decreased performance.
…
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NUMBER OF
INJECTIONS PER
VIAL
This entry defines the number of injections (up to 9) per
vial.
Indicates the injection volume for each injection from
each vial. The maximum programmable injection volume
is:
partial loopfill: 50% of the programmed loop volume
INJECTION VOLUME
µL pick-up: injection volume = (loop volume – 3 x needle
volume)/2
full loop: This parameter is not programmable, and is
equal to the loop volume. It requires more sample to fill
the loop than other techniques.:
3 x loop volume for loop volumes < 100 µL;
2 x loop volume for loop volumes ≥ 100 µL - 499 µL;
1.5 x loop volume for loop volumes ≥ 500 µL.
INJECTION
CONTROL
Note: Injection controls options are available in
Micro mode and only in conjunction with partial
loopfill or full loop injection only.
Determines whether the injection is performed in the is
standard (<STD>) or in the low dispersion mode
(<L.D.>).
FLOW RATE
Enter the flow rate.
L.D. FACTOR
This entry is used to indicate the low dispersion factor,
range 0.7 - 2.0
Note: These two parameters determine the
switching time of the injection valve, refer to
Appendix A for more details.
3.8.11 The Method Menu Item <WASH>
<WASH>
User’s Manual FAMOS
This soft function key is used to access a series of
commands to program wash methods. You can program
a wash between injections, samples or series. When you
select a wash method, a screen will be presented that
will allow you to indicate the volume of wash solvent can
be defined. The minimum programmable volume is
300 µL.
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The User Interface
3.8.12 The Method Menu Item <TIMEBASE>
<TIMEBASE>
(if enabled in System
Menu, Usage Menu)
<AUX>
This soft function key is used to enable control of the
optional ISS valve and other devices via auxiliary (P5) or
binary TTL outputs (P3).
Up to 5 timebase methods can be programmed. The
menu offers the following soft function keys:
This command scrolls through all program lines by
pressing E or select AUX to move to the next auxiliary.
There are four AUX entries (AUX1-AUX4).
<VALVES>
This command controls the ISS valve and the solvent
selection valve. The ISS valve can only be programmed if
the optional ISS valve is installed. The entries 6-1 and 21 refer to the interconnected ports of the valves. Press E
to scroll through programming lines. Enter the time and
the SSV port number (value between 1 and 6).
<CODE>
Enter a time and a hexadecimal value between1 and 15.
Press E to scroll through the programming lines.
<END TIME>
This command is used to enter the end time for timed
events program; press E to scroll through the
programming lines. If no value is filled in or if CL is
pressed, the FAMOS Well Plate Microautosampler will
automatically generate an end time. The end time is
equal to the analysis time programmed in the injection
method used in the same series.
Note: If end time exceeds the programmed
analysis time, this end time overrules the analysis
time. You can program events after the end time,
but these events are not carried out during a run.
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3.8.13 The Method Menu Item <MIX>
If the MIX option is enabled in the System menu, the soft function key <MIX>
will be presented.
<MIX>
This soft function key is used to access a series of
commands to program a method for:
(if enabled in System
Menu, Usage Menu)
• pre-injection sample handling (e.g. pre-column
derivatization)
• dilution
• adding of internal standard
Up to nine mix methods can be programmed. A total of
240 steps can be included in the 9 mix methods and the
user program. Assign a number to the mix method.
When the Mix Menu is presented:
<EDIT>
Press to edit an existing step or a new step for a new
mix method.
<INSERT>
Press to insert a new step in an existing method before
the displayed step.
<DELETE>
Press to delete the displayed step.
Note: “End of mix method” means that the mix
method is empty; if an existing mix method is
selected, the first line of the mix method is
displayed. Scroll through the steps of the existing
method with the cursor keys and use the soft
function keys to enter changes in an existing
method.
The following types of steps can be programmed for a
mix method:
<ASPIRATE>
Aspirates a programmed volume (sample, air, destination,
reagent A-D). Speed of syringe can be selected from 1-9.
(TABLE 3-1). The indicated height (H) is the distance of
the needle point to the plate holder (default: 2 mm).The
maximum amount which can be aspirated is the total
volume of the syringe.
<DISPENSE>
Dispenses (sample, waste, destination, reagent A-D) a
programmed volume from the buffer tubing. Speed of the
syringe can be selected from 1 - 9 (TABLE 3-1).The
indicated height (H) indicated is the distance of the
needle point to the plate holder (default: 2 mm). It is
possible to dispense a larger volume than the volume
aspirated in previous actions. The aspirated amount will
be complemented with liquid from the wash solvent
bottle to total the programmed dispense volume.
...
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The User Interface
<WAIT>
Defines a pause (H:MM:SS, maximum of 9 hours, 59
minutes and 59 seconds).
Note: During the pause, the needle will move to
the home position (if the previous step is an
aspirate or dispense action). If you want the
needle to stay in the same position, an aspirate or
dispense step of 0 µL must be programmed at the
desired position.
<REPEAT>
Enter the number of steps that must be repeated and
how often they must be repeated.
<WASH>
Enter the volume for needle wash. Buffer is rinsed to
waste.
3.8.14 The Method Menu Item <USER PROGRAM>
If the USER PROGRAM option is enabled in the System menu, the soft key
<USER PROGRAM> is presented.
<USER PROGRAM>
The user program offers the possibility to program all
possible actions required for a sample handling sequence
in separate steps.
(if enabled in System
Menu, Usage Menu)
Note: The total number of steps for the user
program and all nine mix methods cannot exceed
240. The user program can be protected by a
special user program protection code (System
Settings, Usage Menu). If no user program has
been programmed yet, “end of user program” is
displayed. Otherwise, the first line of the
programmed method appears.
The following entries are presented for the user program:
<EDIT>
Press to edit an existing step or a new step for a new
mix method.
<INSERT>
Press to insert a new step in an existing method before
the displayed step.
<DELETE>
Press to delete the displayed step.
The edit and insert menus offer the following soft
function keys:
<ASPIRATE>
Removes a programmed volume from sample well,
ambient air, destination vial, wash, or one of the reagent
vials into the buffer tubing. The speed and height of the
syringe can be entered (TABLE 3-3). The maximum
volume that can be aspirated is the total volume of the
syringe.
…
3-22
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User’s Manual FAMOS
The User Interface
<DISPENSE>
Delivers a programmed volume from the buffer tubing
into the sample well, waste, destination vial, wash or
one of the reagent vials. Speed and height of syringe can
be entered (TABLE 3-3).
It is not possible to dispense a larger volume than the
total volume aspirated in previous actions.
This command is used to control the connections of the
syringe to one of its three tubes:
<SYR_VALVE>
<NEEDLE>
to connect the syringe to the sample
needle
<WASH>
to connect the syringe to wash solvent
bottle
<WASTE>
to connect the syringe to the waste
tubing.
This command is used to control the movements of the
syringe.
<SYR>
<WASH>
<LOAD>
the syringe with the programmed volume
<UNLOAD
the syringe with the programmed volume
<HOME>
the volume previously aspirated will be
dispensed to the last programmed
position, and the syringe will be initialized
again.
This command is used to execute a needle wash; the
content of the buffer tubing is not rinsed to waste before
the start of the wash. The programmed volume of wash
solvent is used to wash the needle at the wash position.
Note: The wash position may be contaminated
with the contents of the buffer tubing, which may
generate cross-contamination. To prevent
contamination of the wash position, program a
dispense to waste action before programming a
wash action.
<VALVES>
This command is used to program positions of high
pressure valves (ISS, injector valve, SSV). The injector
valve has two positions: <INJECT> and <LOAD>. The
ISS optional valve has positions indicated by 1-6 (left)
and 1-2 (right).
…
User’s Manual FAMOS
D931R1
3-23
The User Interface
<WAIT>
This command is used to program a pause (max. 9 hours,
59 minutes, 59 seconds).
Note: During the pause, the needle will move to
home position (if the previous step is an aspirate
or dispense action). If you want the needle to
stay in the same position, an aspirate or dispense
step of 0 µL must be programmed at the desired
position.
<COMPRES>
<AUX>
<WAIT-IN>
<PROG-OUT>
<CODE>
<MARKERS>
<SSV>
(option)
3-24
This command is used to activate the compressor to put
air pressure on a sample. The compressor will stay active
until it is switched off (in a next programmed step). The
compressor will be automatically switched off at the end
of the needle wash routine if a needle wash is used.
This command is used to control the four standard
auxiliaries (contact closures). Refer to Appendix E for
details.
This command is used to program a pause in which the
FAMOS Well Plate Microautosampler waits for one of the
four inputs to become <HIGH> or <LOW> before
continuing with the next step. Refer to Appendix E for
details.
This command is used to define two programmable
outputs (contact closures). These are similar to the
auxiliaries, but only available in the user program. Refer
to Appendix E for details.
to program the output to the connector P3 TIMED
OUTPUTS. This is a HEX output in the range 0 to 15.
Refer to Appendix E for details.
the markers normally generated in the FAMOS Well Plate
Microautosampler are not active in the user program, but
can be programmed in this screen (refer to Appendix E
for details). Select marker and status (inject, vial,
labeled).
This command is used to define the Solvent Selection
Valve (SSV) port position, range 1 to 6.
D931R1
User’s Manual FAMOS
The User Interface
TABLE 3-3. Syringe Speed
Speed
Syringe
25 µL
100 µL
250 µL
50
125
500 µL
250
1
12.5
2 (low)
31.3
125
315
630
1.3
3 (normal)
62.5
250
625
1250
2.5
4 (high)
93.8
375
940
1880
3.8
5
192.5
770
1920
3840
7.7
6
267.5
1070
2675
5335
10.7
7
342.5
1370
3430
6855
13.7
8
436.3
1745
4365
8725
17.5
9
533.8
2135
5335
10670
µL/min
CAUTION
1000 µL
0.5
21.3
mL/min
Caution: The pressure in the buffer tubing will increase during the dispense
action. To prevent damage of the buffer tubing, the flow should not exceed the
value of 6mL/min for water. A maximum speed of 9 for 25, 100, and 250 µLsyringes, a maximum speed of 6 for a 500 µL syringe and a maximum speed of 4
for 1000 µL syringe should be used. If more viscous liquids are used, the speeds
should be reduced.
3.8.15 The Method Menu Item <TEMPLATE>
If the TEMPLATE option is enabled in the System menu, the soft function key
<TEMPLATE> is presented.
<TEMPLATE >
Use this key to enter a menu in which the contents of a
template can be defined.
First assign a number to the template, then link the
numbers of methods to the template.
(User program
instead of methods)
<YES>
The following items can be entered to fill a template:
The complete template is filled with the user program; no
other methods can be added.
The template can be filled with the following:
mix method number,
<NO>
injection method number,
wash method number,
timebase method number
A maximum of 24 templates can be programmed
User’s Manual FAMOS
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The User Interface
3.9
Series Menu
The Series Menu allows the user to define the run sequence in a series. A
maximum of 24 series can be programmed, each series contains information
about the methods to be used for a range of wells. This can be a template, a
separate method (mix, injection, wash, timebase), or the user program.
Information on location of wells, labeled wells or calibration wells is also
programmed in a series. Series parameters are described in TABLE 3-4.
Note: The settings entered in the System Menu and the methods defined in the
Methods Menu determine which possibilities appear in the Series Menu.
TABLE 3-4. Series parameters
Without templates
With templates
● Template number
❍ Use user program Yes/No
● Injection method number
● Wash method number
❍Time base methods number
❍ Mix method number
Time base and mix method are only available if enabled in the System Menu
❍
❍
❍
❍
Use calibration wells Yes/No
First calibration well
Last calibration well
No. of samples between calibration
Calibration wells are only available if enabled in the System Menu; not available if Mix
Method has been programmed
● First sample well
● Last sample well
Only if a mix method has been programmed, or if used in user program:
❍ First destination well
❍
❍
❍
❍
Vial
Vial
Vial
Vial
Reagent-A
Reagent-B
Reagent-C
Reagent-D
Only if the use of labeled wells has been enabled in the System Menu:
❍
❍
❍
❍
Labeled
Labeled
Labeled
Labeled
well
well
well
well
no.
no.
no.
no.
1
2
3
4
● marked questions are always asked in series,
❍ marked questions depend on the used methods and the settings entered in the System
Menu.
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User’s Manual FAMOS
The User Interface
After you have entered the required settings in the System Menu and after you
have programmed methods to be used for an analytical run, you can press Series
to enter the Series Menu. TABLE 3-4 gives an overview of the items you should
to define for the Series.
•
With Templates - if you are going to execute an analytical run by way of a
template, you will only be asked to enter the template number and to indicate
the location of the first sample well and the last sample well.
•
Without Templates - if you are going to execute an analytical run without
using a template, you will be asked to enter an injection method number and
a wash method number, and you will have to indicate the location of the first
sample well and last sample well.
If you have enabled use of calibration wells in the System Menu (Usage Menu),
you should define whether you will use calibration wells, and indicate the
location of the first and last calibration well, and indicate the number of wells
between calibration wells (FIGURE 3-6).
However, if you have for example enabled use of a Mix Method in the System
Menu (Usage Menu), you will also have to define the location of the First
destination well and Reagent vials.
Note: Series are stored in the microautosampler memory for as long as the power
is on. As soon as power is switched off, all programmed series will be deleted. It
is not possible to leave the Series Menu before all values have been programmed.
FIGURE 3-6. Injection sequence with 3 calibration wells between every 5 wells
User’s Manual FAMOS
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The User Interface
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3-28
D931R1
User’s Manual FAMOS
Programming Examples
CHAPTER 4
4.1
Overview
The commands described in Chapter 3 can be used by the analyst to generate a
broad range of programs for the operation of the microautosampler. These
commands allow the user to configure the unit to meet the specific needs of the
laboratory. A series of blank forms are provided to assist the analyst in
programming (Appendix F).
This chapter describes the editing of general system settings and selection of the
plate. In addition, a few simple programs that show the general approach to
programming (additional programs are presented in Appendix C) are included.
While these programs may not meet the specific needs of the analyst, it is likely
that they can be used with minor modification (e.g. parameters such as the
analysis time and sample volume may need to be changed). An additional
rationale for the presentation of these programs is to provide a description of the
logical processes that are used in generating a program.
It should be noted that the examples described below could be used with
systems that include UltiMate with the UltiChrom software program as well as
for stand-alone systems.
The following programs are included in this chapter:
• Single 200 nL injection (shows partial loopfill) - Section 4.4.2.
• 100 nL injection from of 3 vials and 5 nL injections from 3 other vials (shows
partial loopfill and injections from multiple vials)- Section4.4.3.
• 1 µL injection out of a 5 µL sample in vial 1 using transport liquid. (shows µL
pickup) – Section 4.4.4.
Additional examples are provided in Appendix C.
User’s Manual FAMOS
D931R1
4-1
Programming Examples
4.2
General System Settings
When the microautosampler is initially installed, a variety of general parameters
are set which generally define the configuration of the unit and should be
checked. When you are defining a new method, you should check these values
to make sure that they are relevant.
To check the general system settings, press the System key, then select the soft
function keys <MICRO> and <GENERAL> as appropriate and press the E key
on a sequential basis to view each selection.
Use the E key to go through the general system settings. A set of reasonable
values is presented in TABLE 4-1.
Note: The E key is used to indicate that the user accepts the indicated value for
a parameter and he/she wants to access the next parameter field. For the sake of
brevity, it will be understood that the E key must be pressed when a parameter is
viewed/edited.
TABLE 4-1. Typical General Parameter Settings
Setting
<GENERAL> Parameter
Volume of the installed injection loop
Volume of the tubing ‘needle – valve’
Syringe speed
Scale factor
Sample needle height
Skip missing vials
Air segment
Headspace pressure
Time base display
Key click
Error beep
Alarm buzzer
5 µL
2.4 µL
low
0.2
04 mm
yes
no
yes
HH:MM:SS or H:MM:mm
on
on
on
To go back to the Ready menu, press Escape twice.
Note: Adjust the needle height to the desired depth of insertion. Use 4mm ([04])
for the enclosed 250 µL polypropylene tapered vials.
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D931R1
User’s Manual FAMOS
Programming Examples
4.3
Installing the Plate
To install the plate:
a) Insert the 48 vial holder (FIGURE 4-1). It will be necessary to use the
<EXCHANGE> soft key to move the plate holder to the left to access the
plate holder.
1
2
3
4
5
6
7
8
A
A
B
B
C
C
D
D
E
E
F
F
1
2
3
4
5
6
7
8
FIGURE 4-1. 48 Position Well Plate
b) Press the <PLATE HOME> soft key to move the plate holder back into
operation position and place a sample vial in position A1.
To check the plates setting:
a) Press the System key and then select the soft function keys <MICRO> and
<PLATES>).
b) Select <48 vials>.
c) Select Process Vials <IN ROWS>.
To go back to the Ready Menu, press Escape twice.
User’s Manual FAMOS
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4-3
Programming Examples
4.4
Sample Applications
4.4.1 Conventions
The following typographical conventions are used:
Initial Capitals
represents the names of the menus (e.g. Ready Menu).
<CAPTALS>
represents the soft function key (e.g. <INJECTION>).
bold type
represents the names of the function keys (e.g. E).
[01.23]
indicates that a numerical value has to be entered
4.4.2 Example 1: Injecting a Single 200 nL Sample from 1 vial
This example describes an application in which a single injection is made from a
single vial in position A1.
a) Place a sample in position A1.
b) Program the Injection Method by entering the parameters indicated in TABLE
4-2.
TABLE 4-2. Programming the Injection Method
Press keys
Methods
<INJECTION> [01] E
<PARTIAL> E
[25.00] E
[5.0] E
[01] E
[02] E
<STD> E
Escape Escape
to
to
to
to
to
to
to
to
to
Description
enter the Methods Menu
define the number of the method
select partial loop fill
set analysis time to 25 min
set the flush volume to 5µL
set the number of injections/vial to 1
set injection volume to 0.2 µL
select the standard injection control
return to the Ready Menu
c) Program the Series by entering the parameters indicated in TABLE 4-3.
TABLE 4-3. Programming the Series:
Press keys
Series
[01] E
[01] E
CL E
<ROW A> [01] E
<ROW A> [01] E
Escape
4-4
to
to
to
to
to
to
to
Description
enter the Series Menu
define the Series number
select the injection method number
enter <NONE> for wash method
define location of the first sample vial
define location of the last sample vial
return to the Ready Menu
D931R1
User’s Manual FAMOS
Programming Examples
d) Run the series by entering the parameters indicated in TABLE 4-4 and
pressing <START>.
TABLE 4-4. Running the Series
Press keys
Start/Stop
[01] E
[01] E
<START>
to
to
to
to
Description
start the FAMOS Well Plate Microautosampler
start at series 01
stop after series 01
start execution of the series.
At the end of the defined analysis time the Ready Menu will appear again to
indicate that the FAMOS Well Plate Microautosampler is ready for the next run.
4.4.3 Example 2: Injecting a Single 100 nL sample from 3 vials and
Injecting 5 µL samples from 3 other vials
This example describes an application in which a single injection is made from
each of three vials and a single injection with a different volume made from three
additional vials.
a) Place the appropriate vials in positions A1-A3 (for the 100 nL Injections) and
the appropriate vials in positions C1-C3 (for the 5 µL injections).
b) Program the injection method for the 100 nL injections by entering the
parameters indicated in TABLE 4-5.
TABLE 4-5. Programming the Injection Method for 100 nL Injections
Press keys
Methods
<INJECTION> [01] E
<PARTIAL> E
[25.00] E
[5.0] E
[01] E
[01] E
<STD> E
Escape
to
to
to
to
to
to
to
to
to
Description
enter the Methods Menu
define 1 for the first method
select partial loop fill
set analysis time to 25 min
set the flush volume to 5µL
set the number of injections/vial to 1
set injection volume to 0.1 µL
select the standard injection control
return to the Methods Menu
c) Program the injection method for the 5 µL injections by entering the
parameters indicated in TABLE 4-6.
TABLE 4-6. Programming the Injection Method for 5 µL Injections
Press keys
<INJECTION> [02] E
<FLUSHED> E
[25.00] E
[5.0] E
[01] E
<STD> E
Escape Escape
User’s Manual FAMOS
to
to
to
to
to
to
to
Description
define 2 for the second method
select flushed loop fill
set analysis time to 25 min
set the flush volume to 5µL
set the number of injections/vial to 1
select the standard injection control
return to the Ready Menu
D931R1
4-5
Programming Examples
d) Program the series by entering the parameters indicated in TABLE 4-7.
TABLE 4-7. Programming the Series
Press keys
Series
[01] E
[01] E
CL E
<ROW A> [01]
<ROW A> [03]
Series
[02] E
[02] E
CL E
<ROW C> [01]
<ROW C> [03]
Escape
E
E
E
E
to
to
to
to
to
to
to
to
to
to
to
to
to
Description
enter the Series Menu
define the Series number
select the injection method number
enter <NONE> for wash method
define location A1 as first sample vial
define location A3 as last sample vial
enter the Series Menu
define the next Series number
select the next injection method number
enter <NONE> for wash method
define location C1 as first sample vial
define location C3 as last sample vial
return to the Ready Menu
e) Run the series by entering the parameters indicated in TABLE 4-8 and
pressing <START>.
TABLE 4-8. Running the Series
Press keys
Start/Stop
[01] E
[02] E
<START>
to
to
to
to
Description
start the FAMOS Well Plate Autosampler
start at series 01
stop after series 02
start execution of the series.
At the end of the defined analysis time, the Ready Menu will appear again to
indicate that the FAMOS Well Plate Microautosampler is ready for the following
next run.
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User’s Manual FAMOS
Programming Examples
4.4.4 Example 3: 1µ
µL injection out of a 5 µL sample in vial A1 using the
transport liquid (µ
µL pick-up)
This example describes an application in which a single injection is made from a
vial position A1 using transport liquid for pickup. The µL pick-up routine allows
for an injection to be made from small sample volumes with essentially no
sample loss.
To perform this operation
a) Place a 250 µL tapered sample vial in vial position A1 and one of the 10 mL
capped vials in transport vial position 1 (leftmost position in the rack) as
shown in FIGURE 4-2 (the transport liquid should have a lower strength than
the mobile phase).
Wash Position
Transport Solvent Positions
1
1
2
3
2
4
3
5
6
4
7
8
A
A
B
B
C
C
D
D
E
E
F
F
1
2
3
4
5
6
7
8
FIGURE 4-2. 48 Well Plate Holder and Location of Transport Liquid/Sample Vials
b) Install a 10µL sample loop.
c) Set the loop volume to 10 µL and verify that the right needle height (4 mm)
has been set so that the needle can reach the bottom of the tapered sample
vial (see Section 4.2).
d) Identify the position of the transport liquid vial by entering the parameters
indicated in TABLE 4-9.
TABLE 4-9. Identifying the Location of the Transport Vial
Press keys
System
<MICRO> E
<PLATES> E
<48 > E
<IN ROWS>
[01] E
[01] E
Escape Escape
User’s Manual FAMOS
Description
to enter the system settings
to select micro mode
to select the right plate version
to
to
to
to
process the well in rows
define the location of the first transport liquid vial
define the location of the last transport liquid vial
return to the Ready Menu
D931R1
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Programming Examples
e) Program the Injection Method by entering the parameters indicated in TABLE
4-10.
TABLE 4-10. Programming the Injection Method
Press keys
Methods
<INJECTION> [01] E
<PICK UP> E
[25.00] E
[1] E
[1.00] E
Escape Escape
f)
Description
to enter the Methods Menu
program injection method number 1
to select partial loopfill injection method
to define an analysis time of 25 minutes
to define the number of injections/vial
to set the injection volume to 1.00 µL
to return to the Ready Menu
Define the Series by entering the parameters indicated in TABLE 4-11.
TABLE 4-11. Programming the Series
Press keys
Series
[01] E
[01] E
CL E
<ROW A> [01] E
<ROW A> [01] E
Escape
to
to
to
to
to
to
to
Description
enter the Series Menu
define the Series number
select the injection method number
enter <NONE> for wash method
define location A1 as first sample vial
define location A1 as last sample vial
return to the Ready Menu
g) Run the series by entering the parameters indicated in TABLE 4-12 and
pressing <START>.
TABLE 4-12. Running the Series
Press keys
Start/Stop
[01] E
[01] E
<START>
to
to
to
to
Description
start the FAMOS Well Plate Microautosampler
start at series 01
stop after series 01
start execution of the series.
At the end of the defined analysis time the Ready Menu will appear again to
indicate that the FAMOS Well Plate Microautosampler is ready for the following
next run.
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Testing the Microautosampler
CHAPTER 5
5.1
Overview
This chapter describes a series of activities that can be used to check the
operation of the system and to verify that your FAMOS Well Plate
Microautosampler is operating in an acceptable manner.
This chapter includes:
•
Establishing a Test Protocol (Section 5.2)
•
Running the Test Protocol (Section 5.3)
User’s Manual FAMOS
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5-1
Testing The Microautosampler
5.2
Establishing a Test Protocol
This section describes the parameters for the test protocol that will be performed
in Section 5.3. A detailed discussion of how the parameters are accessed and
edited is presented in Chapter 3.
a) Verify that the microautosampler is configured as listed in TABLE 5-1 and set
the values in TABLE 5-1 for the General Parameters.
TABLE 5-1. General Parameters
Description
Loop volume
Needle tube volume
Syringe volume
Dispenser speed
Sample needle height
Skip missing vial
Air segment
Head space pressure
Value
5 µL
2.5 µL
25 µL
Low/0.2
2 mm
YES
NO
YES
b) Set the well plate type you are using.
c) Program the Partial Loopfill method presented in TABLE 5-2.
TABLE 5-2. Method Parameters
Description
Injection method number
Analyze time
Flush volume
Injections/vial
Injection volume
Low dispersion mode
Value
1
1:00 min
5 µL
9
0.10 µL
off
d) Program the Series presented in TABLE 5-3.
TABLE 5-3. Series Parameters
Description
Series number
Injection method number
Wash method
First vial
Last vial
5-2
Value
1
1
None
ROW A01
ROW A01
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User’s Manual FAMOS
Testing The Microautosampler
5.3
The Test Protocol
5.3.1 The Analytical System used for the Test Protocol
In this test protocol, a LC Packings UltiMate Micropump (with a CAP-300
calibrator) and an UltiMate UV Detector, set at 254 nm are used (FIGURE 5-1).
The UltiChrom software is used to control the system and acquire/present the
data. A 1 m fused silica tube (75 µm I.D.) should be placed between the
autosampler and the detector to create backpressure.
FIGURE 5-1. Analytical System for Test Protocol
If an UltiMate system is not available, any HPLC system with reproducible
microflow capabilities can be used (e.g. a conventional HPLC pump with an
Acurate flow splitter).
Note: The entire flow path from the syringe to the needle should be carefully
inspected before the test protocol is performed. This test should not be
performed if the ambient temperature is below 18oC.
5.3.2 The Test Protocol
Prepare the FAMOS Well Plate Microautosampler as follows:
•
Place a closed vial at position A01, the sample is Uracil in distilled water
~0.1 mg/mL The eluent is distilled water.
•
Fill the wash solvent bottle with 80% H2O / 20% isopropanol.
•
Program a partial loop fill method using the settings in Section 5.2.
Obtain the peak area for each peak and determine the value for σ, as well as the
RSD%, reproducibility, which should be ≤1.0% using the equations presented
below.
Peak area =
σ n −1 =
∑ Peak area
n
∑ ( Peak area
−
Peak area
n −1
σ n −1
RSD% =
× 100%
Peak area
User’s Manual FAMOS
D931R1
)
2
5-3
Testing The Microautosampler
A typical chromatogram from the test is presented in FIGURE 5-2 and sample
data is presented in TABLE 5-4.
FIGURE 5-2. Typical Sample Chromatogram for the reproducibility test
TABLE 5-4. Sample Peak Height Data
Peak number
1
2
3
4
5
6
7
8
9
Height
216.8
259.6
262.8
264.4
265.0
263.0
263.4
262.9
265.3
For this example, the results are:
RSD% = 0.66 %
The specified value is an RSD% < 1%.
If the reproducibility is not within the specification:
a) Check for any air bubbles in the tubing/syringe.
b) Check needle and tube connections between needle and injection valve for
dead volume or blockage.
c) Review the Troubleshooting Section (Section 6.3).
Once you have remedied the problem, repeat the performance test.
5-4
D931R1
User’s Manual FAMOS
Testing The Microautosampler
5.4
Checking System Components
5.4.1 Testing the Accuracy of Delivery of the Syringe
To determine the accuracy of delivery of the syringe, dispense a volume of 25 µL
of water from a sample vial to a destination vial, program the following Mix
Method (TABLE 5-5) and Series (TABLE 5-6).
TABLE 5-5. Mix Method 1
Step
1
2
3
Action
Aspirate 25 µL Sample
Dispense 25 µL to Destination
End of mix method
Speed
2
3
Height
05
03
TABLE 5-6. Series 1
Description
Series Number
Injection method
Wash method
Mix method
First well
Last well
First destination vial
Reagent A vial
Value
1
None
None
1
ROW A 01
ROW A 01
ROW A 02
1
To determine the accuracy of delivery of the syringe:
a) Weigh the destination vial before and after the Run.
b) The difference is the aspirated volume
The specified variation is ± 2%.
5.4.2 Testing the Loop Volume
To determine the volume of the injection loop
a) Disconnect the loop from the injection valve.
b) Remove all liquids from the loop with air.
c) Weight the empty loop on an analytical balance.
d) Fill the loop with minimal 2 times its volume of water.
e) Weight the filled loop again.
The difference in the weight of the loop, divided by the specific weight of water
(1 g/mL) gives the calibrated volume of the loop.
The specified variation is ± 10%.
User’s Manual FAMOS
D931R1
5-5
Testing The Microautosampler
5.4.3 Testing the Tray Cooling Option
To test the tray cooling option (if installed):
a) Remove the plate holder.
b) Place a thermocouple in the middle of the cooling plate (flat surface
underneath the wash solvent bottle, item 3, FIGURE 2-5), making sure that a
good contact is made.
c) Switch on the cooling and program a setpoint of 10°C.
d) Wait minimal 15 minutes for equilibration of the FAMOS Well Plate
Microautosampler.
e) Read out the temperature of the thermocouple.
The value must be in a range of ± 2°C of the programmed setpoint.
Note: When the cooling option is used, the temperature of the sample inside
the sample vial may be slightly different than the temperature set via the
command, due to a variety of effects (e.g. the heat transfer
characteristics of the vial walls). If it is necessary to precisely set the
temperature of the sample, we recommend that you determine the temperature
inside the vial at various temperature setpoints and set the cooling option
so that the desired internal temperature is attained.
5-6
D931R1
User’s Manual FAMOS
Maintenance and
Troubleshooting
CHAPTER 6
6.1
Overview
This chapter provides information to assist in optimizing the performance of the
FAMOS Well Plate Microautosampler and maintaining it in your laboratory. It
includes the following material:
•
Maintenance - describes a series of activities that should be performed on a
periodic basis to optimize the performance of the system and minimize down
time (Section 6.2).
•
Replacing Components – provides directions for replacing components due to
wear or to re-configure the system (e.g. changing the syringe) to meet the
requirements of a different analytical procedure (Section 6.3).
•
Troubleshooting - discusses a series of activities that should be used to
determine the cause of a problem. (Section 6.4).
•
Error Codes - lists and describes various messages that are presented on the
display to indicate a fault with the system (Section 6.5).
•
Spare Parts/Replacement Parts Lists – Presents a listing of components that
are used to maintain the unit or to change the configuration (Section 6.6).
User’s Manual FAMOS
D931R1
6-1
Maintenance and Troubleshooting
6.2
Maintenance
Maintenance refers to a variety of activities that should be performed on a
routine basis to optimize performance of the system. Many routine maintenance
activities can be readily performed by the user.
In some cases (e.g. replacement of critical components), we recommend that a
factory trained service engineer should be called to perform the operation. This
will ensure optimal long term performance and maximum uptime. LC Packings
provides a broad range of service support activities to ensure that the FAMOS
Well Plate Microautosampler is functioning in a suitable manner. These activities
can be customized to meet the specific needs of the customer. For further
information, please contact your local LC Packings office or representative.
TABLE 6-1. Recommended Maintenance Schedule
Frequency
Every Day
Every 3 months
Every year
Operation
Before operating, check for any air bubbles in
the fluidic lines and degas the wash solvent.
Check that there are no leaks of the fluidics
connections.
Check that salts are not deposited by the
fluidics joints.
When using buffer solutions, flush the system
thoroughly after use with a solvent that contains
does not contain buffers/salts.
Inspect the condition of all tubing (cracks, nicks,
cuts, clogging).
Replace:
Rotor Seal
Sample Needle
Prepuncturing Needle
Syringe tip
Connections on Injection Valve and Syringe
Check:
Stator
Note: The frequency of the various activities described above is a good starting
point. As the user gains experience with the system, it will be found that some
activities can be done less frequently and other need to be done more frequently.
The frequency is dependent on a number of factors including the nature of the
sample and the mobile phase.
6-2
D931R1
User’s Manual FAMOS
Maintenance and Troubleshooting
6.3
Replacing Major Components
A variety of components on the FAMOS Well Plate Microautosampler can be
readily changed by the user as required to ensure that the instrument is
maintained in optimal condition. In some circumstances the analyst may want to
change a component, as an example, four different needles are available (see
Section 6.6.2) and it is possible that the application would be best served with a
silica coated needle if the sample could interact with the stainless steel needle.
6.3.1 Replacing the Syringe
The FAMOS Well Plate Microautosampler is supplied with a 25 µL syringe
(standard configuration). In addition, a 100, 250, 500 or 1000 µL syringe is
available.
To replace the syringe:
a) Press the <SYR END> soft function key in the Ready Menu to move the
syringe to the end position, and then lift the cover.
b) Unscrew the top of the syringe (turn clockwise). Pull the syringe down until
it releases from the syringe valve. Then pull the syringe towards you and
remove it (FIGURE 6-1).
c) Unscrew the Luer Lock adapter (100, 250, 500 or 1000 µL syringe only).
Syringe Valve
Luer Lock
Adapter *)
Syringe
Syringe Tip
Syringe Shaft
*) 100, 250, 500, 1000 uL syringes only
FIGURE 6-1. Replacing the syringe
d) Attach the Luer Lock adapter to the new syringe (100, 250, 500 or 1000 µL
syringe only).
Note: The 100, 250, 500 and 1000 µL syringes require the Luer Lock adapter
(P/N 160137).
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6-3
Maintenance and Troubleshooting
e) Fill the new syringe with wash solvent and make sure that all air bubbles are
removed from the syringe. Connect the bottom of the filled syringe to the
FAMOS Well Plate Microautosampler. Screw the top of the filled syringe to
microautosampler (counter clockwise).
f)
CAUTION
Lower the cover. Press the <SYR HOME> soft function key to remove air
from the syringe. The syringe moves to the home position and the content is
dispensed to waste.
Caution: Before continuing, check if the correct syringe is selected in the System
Menu and the correct buffer tubing is installed (refer to Section 2.3). An
incorrect setting may lead to damage to the system.
g) Select the <WASH>soft key in the Ready Menu to execute a standard wash
routine. All tubing connected to the syringe valve is filled and rinsed.
6.3.2 Replacing the Syringe Tip
To replace the tip of the syringe:
a) Remove the syringe as described in Section 6.3.1, items a-c.
b) Remove the syringe tip.
c) Place a new tip on a flat surface (e.g. workbench), making certain that
attachment point is facing up.
d) Gently push the syringe shaft in the new tip.
CAUTION
Caution: If you are installing a 25 µL syringe tip, be extremely careful as the shaft
is very thin and can be easily bent.
e) Carefully insert the shaft in the syringe body.
f)
Re-install and flush the syringe as described in Section 6.3.1 item d) to g).
g) Check for any leakage.
6.3.3 Needle Assembly
The sampling needle consists of two parts:
6-4
•
Sample needle: placed inside the hollow prepuncturing needle; used for the
actual transport of sample. Different types of needles can be used (see
Section 6.6.2). If a needle with different diameter is used, a different air
outlet nut (item 6, FIGURE 6-2) must be used that matches the injection
needle. The fused silica needle (P/N 160116) is the default needle.
•
Prepuncturing needle: a hollow needle used for puncturing of the septum,
capmat or sealer; also used to put headspace pressure on the sample
(approximately 0.5 bar).
D931R1
User’s Manual FAMOS
Maintenance and Troubleshooting
Note: Most commercially available sealers or capmats cannot be used in
combination with headspace pressure. We recommend that headspace pressure
is switched off in those cases (System Menu, General Menu).
A schematic diagram of the needle assembly is shown in FIGURE 6-2 and a list
of the optional needles is presented in TABLE 6-2 and TABLE 6-3.
1 Nut and Ferrule *)
2 Needle Tubing *)
3 Needle Connection Nut *)
4 Conventional Sample Needle *)
5 Needle Holder
6 Air Outlet Nut *)
7 Prepuncturing Needle
8 Sensor
*) constitute the sample needle
FIGURE 6-2. Needle Assembly
TABLE 6-2. Optional Needles
Needle Type
Fused silica needle
Stainless steel needle
Extended needles
Description
needle with small inner diameter for small injection
volumes
sample needle with large inner diameter for viscous
samples, or in case large volumes are loaded in the loop
needles for switching valve placed in the side panel of
the FAMOS Well Plate Microautosampler
Refer to Section 6.6 for an overview of options available for the FAMOS Well Plate
Microautosampler
To replace the present needle:
a) Loosen the needle connection nut (item 3, FIGURE 6-2).
b) Loosen the VICI-Valco and ferrule nut (item 1, FIGURE 6-2).
c) Carefully pull out sample needle and tubing.
d) Remove the air outlet nut and replace it by a new one.
e) Insert a new sample needle and tube through the needle holder and tighten
the nut.
User’s Manual FAMOS
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6-5
Maintenance and Troubleshooting
f)
Connect the other end of the tube to port 4 of the VICI-Valco injection valve
using a VICI-Valco nut and ferrule (item 1, FIGURE 6-2). Do not overtighten
(to prevent block of tubing).
g) Lower the cover of the FAMOS Well Plate Microautosampler.
h) Check sample needle height (default height: 2 mm from plate). If necessary,
adjust the value in the System Menu (General Menu; refer to Chapter 3).
i)
Select the <WASH> soft key in the Ready Menu to clean the new sample
needle.
To install a different needle:
a) Remove needle as described above.
b) Remove the standard air outlet nut (Item 6, FIGURE 2-3, item 6) and replace
it by the nut supplied with the optional sample needle.
c) Install the optional sample needle as described in Section 6.4.3.
d) Adjust settings in System Menu (General Menu) to the volume of the new
needle tubing (see section User Interface).
TABLE 6-3. Characteristics of Needles for FAMOS Well Plate Microautosampler
Needle
O.D.
(mm)
Fused Silica Needles
160116 / 162000 a) 0.280
160117
b) 0.375
Stainless Steel Needles
160119
0.64
contact LC Packings
b) 0.64
Part Number
I.D.
(mm)
Length O.D.
(mm) (mm)
0.10
0.15
300
585
0.25
0.25
135
135
Tubing
Total
I.D.
Length Volume
(mm)
(mm)
2.4 µL
10.3 µL
one piece
1.6
1.6
0.25
0.25
140
420
15 µL
30 µL
a) inert version, b) extra long version
Refer to Section 6.6 for an overview of options available
for the FAMOS Well Plate Microautosampler
Note: When the injection valve is mounted on the right side, e.g. using the
FAMOS Well Plate Microautosampler in combination with the THERMOS
, the
sample needle should be an extra long version.
6.3.4 Prepuncturing Needle
To replace the prepuncturing needle:
a) Remove sample needle (Section 6.3.3).
b) Unscrew the prepuncturing needle.
c) Install a new needle using a new seal.
d) Reinstall the sample needle (Section 6.3.3).
6-6
D931R1
User’s Manual FAMOS
Maintenance and Troubleshooting
6.3.5 Combination Syringe, Sample Loop and Buffer Tubing
This section will indicate the standard configuration of the FAMOS Well Plate
Microautosampler and appropriate configuration for three commonly selected
modes of using the microautosampler (a detailed discussion of the injection
principles is presented in Appendix A):
- Injection volumes smaller than 1 µL
- Injection volumes up to twice the standard (conventional mode)
- For volumes larger than 200 µL
A) Standard Configuration
The standard configuration for the FAMOS Well Plate Microautosampler includes
a 2.4 µL Fused Silica Needle, a 25 µL syringe; a 50 µL buffer tubing and a 10 µL
sample loop. The injection volume ranges shown in TABLE 6-4 are available for
the various injection modes while TABLE 6-5 presents the maximum injection
volumes for the various injection modes. Five sizes of syringes can be used in the
FAMOS Well Plate Microautosampler, 25, 100, 250, 500 and 1000 µL.
TABLE 6-4. Injection Ranges
Injection Mode
Injection Volume Range
Maximal Injection Volume
Full loop
10 µL
loop volume (injection volume fixed
by loop size)
Partial loopfill
0.01 - 5 µL
50% of loop volume
µL pick-up
0.01 – 1.4 µL
(loop volume - 3 x needle tubing
volume) / 2
The characteristics of each injection mode is described below:
•
Full loop injection - gives maximum reproducibility (RSD < 0.4%).
•
Partial loopfill - gives maximum accuracy (depends on syringe accuracy) and
reproducibility better than 0.6% RSD for injection volumes > 1 µL.
•
µL Pick-up - offers zero sample loss, maximum accuracy (same as partial
loopfill) and RSD better than 1.5% for injection volumes ≥ 1 µL.
A discussion of the appropriate modes for three common scenarios is presented
below.
B) Injection Volumes Smaller than 1 µL
Partial loopfill: For maximum reproducibility and accuracy a 25 µL syringe and a
1 µL sample loop should be used.
µL Pick-up: For optimum accuracy and reproducibility a 25 µL syringe and a 10
µL sample loop should be used. The sample plug is transported into the loop,
preceded by a plug of transport liquid that has a volume of 2.5 times the
programmed needle tubing volume.
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D931R1
6-7
Maintenance and Troubleshooting
C) Injection Volumes up to Twice the Standard
With a 100 µL syringe, a stainless steel needle with tubing (15 µL) and 500 µL
buffer and a 50 µL sample loop, the maximum injection volumes are shown in
TABLE 6-5.
TABLE 6-5. Maximum Injection Volume
Injection Mode
Maximal Injection Volume
Full loop
50 µL (sample loss equals 130 µL; two loop volumes
overfill; 30 µL pre-flush)
Partial loopfill
25 µL
µL pick- up
2.5 µL
D) For Volumes Larger than 200 µL
With the 2000 µL buffer tubing, use the appropriate sample loop size and the
appropriate syringe. The syringe volume should be at least 2 x injection volume.
Note: Injection volumes larger than 500 µL are possible, but the sample may
contaminate the syringe. Make certain that you have programmed a sufficient
wash after use
6.3.6 Replacing the Main Fuses and Setting the Operation Voltage
DANGER
Danger: Disconnect the instrument from the electrical supplies before
inspecting/changing the fuse.
To change the Fuses:
a) Pull out the fuse holder (FIGURE 6-3, item 1).
2
1
Orientation
Marks
FIGURE 6-3. The Fuse Compartment
b) Replace the blown fuse(s) by fuse(s) of the same rating:
115 V (AC) ± 10%:
two 5 AT fuses (slow, ¼” x 1¼”, UL/CSA)
230 V (AC) ± 10%:
two 2.5 AT fuses (slow, 5 x 20 mm, IEC127)
(The fuses used are UL-listed and CSA-certified)
6-8
D931R1
User’s Manual FAMOS
Maintenance and Troubleshooting
c) Re-install the fuse holder.
CAUTION
Caution: Make certain that the fuse holder is inserted in the correct position so
that the appropriate voltage setting is indicated (FIGURE 6-3 indicates the
configuration for 220-240 V).
User’s Manual FAMOS
D931R1
6-9
Maintenance and Troubleshooting
6.4
Troubleshooting
Troubleshooting refers to the determination of the cause of a problem. Since the
FAMOS Well Plate Microautosampler is incorporated into an HPLC system, the
first step to determine if the problem is due to the microautosampler is to remove
the unit from the system, install a manual injector and then perform an injection
and compare the results from the two runs. If the results are fine, the problem is
due to the autosampler.
Analytical problems also might be caused by external influences, like temperature
and/or light sensitive samples. For this reason it is important to be sure the
application was running without problems before and nothing has been changed.
A chart outlining the most common faults and the remedies is presented in
FIGURE 6-4.
BAD
REPRODUCIBILITY
Air in Flow
Path
Start a manual wash
Leaking
Syringe
Replace plunger tip or
syringe
Leaking
Syringe Valve
Replace Valve
Rotor seal
worn out
Replace rotor seal
Check Stator face
Dead volumes
in tubing
connections
Redo connections with
new ferrules and nuts
FIGURE 6-4. Troubleshooting the Microautosampler
Note: It is important to note that analytical problems might be caused by external
influences, such as the temperature and or the analysis of light sensitive
samples. When troubleshooting, make certain that the application was running in
an acceptable fashion before the problems were observed and that nothing has
been changed in the application.
6-10
D931R1
User’s Manual FAMOS
Maintenance and Troubleshooting
6.5
Error Codes of the Microautosampler
The FAMOS Well Plate Microautosampler will display an error message if the
user tries to enter invalid data and information on the allowed range will be
displayed. In addition, several sensors are installed to check for possible
mechanical failures and protect the instrument from severe damage.
If a fault or mechanical failure is observed during the operation of the FAMOS
Well Plate Microautosampler, an error code will be presented on the display.
When a message is indicated, press the Start/Stop key twice to remove the
message and correct the fault.
The error codes are indicated in TABLE 6-6 to TABLE 6-11.
TABLE 6-6. Error Codes for Injection Valve and ISS Unit
Error Code
ERROR 11
ERROR 12
ERROR 13
ERROR 14
ERROR 15
Probable Cause
Injection valve is not in a valid position.
The injection valve did not switch within 1.5
seconds.
The switching time of the injection valve
exceeds 500 msec.
ISS valve is not in a valid position.
The ISS valve did not switch within 1.5
seconds.
Solution
Contact
LC Packings.
Contact
LC Packings.
Contact
LC Packings.
Contact
LC Packings.
Contact
LC Packings.
TABLE 6-7. Error Codes for Syringe Dispenser Unit
Error Code
ERROR 21
The syringe valve did not switch.
Probable Cause
ERROR 22
The syringe did not reach home position in time.
ERROR 23
ERROR 24
The syringe spindle did not make the correct
number of rotations.
The spindle does not rotate.
ERROR 25
The syringe valve did not find a valid position.
Solution
Contact
LC Packings.
Contact
LC Packings.
Contact
LC Packings.
Contact
LC Packings.
Contact
LC Packings.
TABLE 6-8. Error Codes for Plate and Plate Holder
Error Code
ERROR 59
ERROR 90
Probable Cause
ERROR 92
Missing plate.
Plate home time-out, plate did not reach home
position (home error).
Plate did not reach or leave home position during
run.
Plate holder missing.
ERROR 93
Plate holder movement is blocked.
ERROR 91
User’s Manual FAMOS
D931R1
Solution
Check for Plate.
Reinstall Plate
Holder.
Reinstall Plate
Holder.
Check/reinstall
Plate Holder.
Check for
blockage.
6-11
Maintenance and Troubleshooting
TABLE 6-9. Error Codes for Injection needle unit
Error Code
ERROR 30
ERROR 31
ERROR 32
ERROR 34
ERROR 39
ERROR 40
ERROR 41
ERROR 42
ERROR 53
Probable Cause
Solution
The sample needle arm did not reach or leave
home position (vertical).
The sample needle arm is in an invalid horizontal
position while moving down.
The sample needle arm did not reach or leave
destination within a certain time (horizontal).
Sample needle arm not in vertical home position
while moving horizontally.
Vial sensor sticks
The sample needle spindle does not rotate
correctly.
The sample needle did not reach or leave home
position.
The sample needle is not at home position.
The sample needle arm is not in the home
position while moving the plate.
Contact
LC Packings.
Contact
LC Packings.
Contact
LC Packings.
Contact
LC Packings.
Contact
LC Packings.
Contact
LC Packings.
Contact
LC Packings.
Contact
LC Packings.
Contact
LC Packings.
TABLE 6-10. Error Codes for Vials
Error Code
ERROR 60
Probable Cause
ERROR 62
Missing vial. Only available when Skip Missing
Vial is set to NO in the System Settings and
during the execution of the Mix of a sample on
48-vial plate.
Missing transport vial.
ERROR 64
Missing vial for reagent A.
ERROR 65
Missing vial for reagent B.
ERROR 66
Missing vial for reagent C.
ERROR 67
Missing vial for reagent D.
ERROR 68
Missing destination vial.
ERROR 69
Not enough transport liquid available due to
missing transport vials.
Solution
Check/install the
vial.
Check/install
vial.
Check/install
vial.
Check/install
vial.
Check/install
vial.
Check/install
vial.
Check/install
vial.
Check/install
vial.
the
the
the
the
the
the
the
TABLE 6-11. Error Codes for Electronics
Error Code
ERROR 71
Solution
Contact
LC Packings
Contact
LC Packings
ERROR 73
Current limit of the external I/O exceeded.
ERROR 75
Error occurred during initialization, the FAMOS
Well Plate Microautosampler cannot start.
Contact
LC Packings
Contact
LC Packings
ERROR 72
6-12
Probable Cause
Flex PCB of the sample needle is not properly
connected.
Invalid configuration of the FAMOS Well Plate
Microautosampler, PCB missing.
D931R1
User’s Manual FAMOS
Maintenance and Troubleshooting
6.6
Spare Parts List
6.6.1 Major Items
Part Number
Description
Standard Version (Stainless Steel Injection Valve)
160105
161168
161170
161171
FAMOS Well Plate Microautosampler
FAMOS Well Plate Microautosampler,
with Cooling option 10 – 40 °C installed
FAMOS Well Plate Microautosampler,
with Feeder option installed
FAMOS Well Plate Microautosampler,
with Feeder and Cooling option 10 – 40 °C installed
Inert Version (PAEK Injection Valve)
160614
161169
161172
161173
FAMOS Well Plate Microautosampler, inert version
FAMOS Well Plate Microautosampler, inert version,
with Cooling option 10 – 40 °C installed
FAMOS Well Plate Microautosampler, inert version,
with Feeder option installed
FAMOS Well Plate Microautosampler, inert version,
with Feeder and Cooling option 10 – 40 °C installed
Firmware
160108
Firmware, ERPOM and RAMs for FAMOS Well Plate
6.6.2 Needle Assembly
Part Number
160152
160153
160120
162019
Description
Air/pre-puncturing needle, including seal
Needle guide body
Needle guide / Air nut for Fused Silica Needle
Needle guide / Air nut for Conventional Needle
Standard Version (Stainless Steel Injection Valve) - Micro
Mode
160116
160117
Fused silica injection (sample) needle, 100 µm I.D., pre-assembled
(2.4 µL).
Fused silica fraction/injection needle, 75 µm I.D., extra long
version (10.3 µL)
Standard Version (Stainless Steel Injection Valve) Conventional Mode
160119
Standard (conventional) stainless steel sample needle, supplied
with TEFZEL tubing (15µL)
Inert Version (PAEK Injection Valve) - Micro Mode
162000
Fused silica injection (sample) needle, 100 µm I.D., pre-assembled
(2.4 µL), for inert injection valve
Inert Version (PAEK Injection Valve) - Conventional Mode
161474
User’s Manual FAMOS
Inert conventional needle for FAMOS, made of PEEK, for inert
injection valve
D931R1
6-13
Maintenance and Troubleshooting
6.6.3 Syringe and Buffer Tubing
Part Number
160128
160129
160130
160131
160132
160137
160155
160156
160157
160158
160159
162020
162021
162022
160161
160160
Description
Syringe 25 µL
Syringe 100 µL
Syringe 250 µL
Syringe 500 µL
Syringe 1 mL
Luer lock adaptor for FAMOS Well Plate Microautosampler
Plunger replacement tip 25 µL (Set of 10)
Plunger replacement tip 100 µL (Set of 10)
Plunger replacement tip 250 µL (Set of 10)
Plunger replacement kit 500 µL (Set of 10)
Plunger replacement kit 1000 µL (Set of 10)
Syringe waste tubing for FAMOS
Syringe wash tubing for FAMOS
Tubing SSV to syringe valve for FAMOS
Flangeless ferrule 1/16” blue for “buffer” tubing connection to low
dispensing valve
Male nut 1/16” blue for “buffer” tubing connection to low
pressure dispensing valve
Standard Version (Stainless Steel Injection Valve)
160125
160126
160127
160143
160142
160141
160140
Buffer tubing 50 µL
Buffer tubing 500 µL
Buffer tubing 2000 µL
Upgrade kit for FAMOS,
Upgrade kit for FAMOS,
Upgrade kit for FAMOS,
Upgrade kit for FAMOS,
for
for
for
for
100 µL syringe
250 µL syringe
500 µL syringe
1.0 mL syringe
(b)
(b)
(b)
(b)
Inert Version (PAEK Injection Valve)
161018
161264
162023
162024
162025
162026
162027
Buffer tubing 50 µL, for inert injection valve
Buffer tubing 500 µL, for inert injection valve
Buffer tubing 2000 µL, for inert injection valve
Upgrade kit for FAMOS, for 100 µL syringe, inert
Upgrade kit for FAMOS, for 250 µL syringe, inert
Upgrade kit for FAMOS, for 500 µL syringe, inert
Upgrade kit for FAMOS, for 1.0 mL syringe, inert
version
version
version
version
(b)
(b)
(b)
(b)
(b) includes Luer lock adapter (P/N 160137), syringe and buffer tubing
6-14
D931R1
User’s Manual FAMOS
Maintenance and Troubleshooting
6.6.4 Injection Valve
Part Number
Description
Standard Version (Stainless Steel Injection Valve)
160163
160149
160151
160109
160110
160111
160112
160113
160114
161481
161482
161483
160115
VALCO C2, 6-Port Micro Injection Valve ASSY
Replacement rotor for VALCO C2, 6-Port Micro Injection Valve
Stator VALCO C2, 6-Port Micro Injection Valve
1 µL loop, FAMOS, PEEK shielded fused silica tubing
5 µL loop, FAMOS, PEEK shielded fused silica tubing
10 µL loop, FAMOS, PEEK shielded fused silica tubing
20 µL loop, FAMOS, PEEK shielded fused silica tubing
50 µL loop, FAMOS, stainless steel tubing
100 µL loop, FAMOS, stainless steel tubing
125 µL loop, FAMOS, stainless steel tubing
250 µL loop, FAMOS, stainless steel tubing
500 µL loop, FAMOS, stainless steel tubing
Connecting tubing valve/micro column, 50 µl I.D., including low
dispersion union
Inert Version (PAEK Injection Valve)
161265
161003
161004
161015
161016
161017
162028
162035
162029
161263
162030
162031
161499
VALCO C2, 6-Port PAEK Micro Injection Valve ASSY
Replacement rotor for VALCO C2, 6-Port Micro Injection Valve,
inert version
PAEK Stator for VALCO C2, 6-Port Micro Injection Valve, inert
version
1 µL loop for FAMOS inert, PEEK shielded fused silica tubing
5 µL loop for FAMOS inert, PEEK shielded fused silica tubing
10 µL loop for FAMOS inert, PEEK shielded fused silica tubing
20 µL loop for FAMOS inert, PEEK shielded fused silica tubing
50 µL loop for FAMOS inert, PEEK tubing
100 µL loop for FAMOS inert, PEEK tubing
125 µL loop for FAMOS inert, PEEK tubing
250 µL loop for FAMOS inert, PEEK tubing
500 µL loop for FAMOS inert, PEEK tubing
Connecting tubing valve/micro column, 50 µl I.D., including low
dispersion union, for inert injection valve
6.6.6 Vials and Accessories
Part Number
162034
162032
160134
160133
161485
160136
160135
161484
161493
161494
160162
162033
User’s Manual FAMOS
Description
Wash vial for FAMOS
Transport Vial 10 mL (20 x 45), 4 PCS
Polypropylene vials for FAMOS, 250 µL, 1000 PCS
Polypropylene vials for FAMOS, 250 µL, 100 PCS
Polypropylene vials for FAMOS, 250 µL, 50 PCS
Polypropylene caps for FAMOS, 1000 PCS
Polypropylene caps for FAMOS, 100 PCS
Polypropylene caps for FAMOS, 50 PCS
Glass vials, 1.5 mL, 100 PCS
Glass inserts, 200 µL, 100 PCS
Wash solvent bottle 100 mL
Wash solvent bottle 100 mL
D931R1
6-15
Maintenance and Troubleshooting
[This page intentionally left blank]
6-16
D931R1
User’s Manual FAMOS
Specifications
CHAPTER 7
7.1
General
7.1.1 Analytical Performance
Capped and
sealed vials
full loop injections
partial loopfill
injections
µL pick-up
injections
Open vials/plates
[η=1.0 cP]:
full loop injections
partial loopfill
injections
µL pick-up
injections
Memory effect
User’s Manual FAMOS
Reproducibility
RSD ≤ 0.4 %
RSD ≤ 0.6 %, injection volumes ≥ 1 µL, with headspace
pressure on the vial and 5 µL pre-flush
RSD ≤ 1.5 %, injection volumes ≥ 1 µL, without
headspace pressure on the vial
Reproducibility
RSD ≤ 0.3 %
RSD ≤ 1.0%; injection volumes ≥ 1 µL, without
headspace pressure on the well.
RSD ≤1.5 %; injection volumes ≥ 1 µL, without
headspace pressure on the vial.
< 0.01 % with programmable needle wash
D931R1
7-1
Specifications
7.1.2 Programming
Full loop injections
Partial loopfill injections
µL pick-up injections
0.01 µL - 1000 µL, full loop, depending on system settings
0.01 - 500 µL, with 0.1 µL increments for partial loopfill
0.1 µL - max. volume, with 0.01 µL increments for µL pickup
max. volume = ( (loop volume - 3 x needle volume) /2)
Max. 9 (volumes are programmable for each injection)
Max. 9 hrs. 59 min. 59 sec.
Programmable (between injections, wells or series)
Freely programmable
Freely programmable, 24 series max
Injection methods
Injection volume
Injections per vial
Analysis time
Needle wash
Priority sample
Series
7.1.3 Inputs/Outputs
Digital Output
Analog Input
Analog Output
2 RS-232
2 EVENT: (Open Collector, Relay)
START-, ERROR IN: Optocoupler
START-, ERROR OUT: Open Collector
1 V (1V/100 nm)
2 ±0.1 V/ ±1 V / ±10 V (for recorder or integrator)
7.1.4 Physical
Dimensions
(WxDxH)
Weight
280 mm (11.0 in) x 400 mm (15.7 in) x 440 mm (17.3 in).
23 kg (50.7 lb.).
7.1.5 Electrical
Power
Requirements
Fuses
7-2
115 VAC; + 15/-20 %; 50 Hz/60 Hz; 250 VA
230 VAC; + 15/-20 %; 50 Hz/60 Hz; 250 VA
115 VAC; two 5.0 AT-fuses; (¼” x ¼”, UL/CSA)
230 VAC; two 2.5 AT-fuses; (5 x 20 mm, IEC 127)
All fuses UL-listed and CSA-certified
D931R1
User’s Manual FAMOS
Specifications
7.1.6 Communication
Outputs
Inputs
Serial
Communication
Inject marker
Well marker
Labeled well marker
Stop I/O
4 Auxiliary outputs
2 Programmable outputs
Alarm output
4 Bit timebase
Next injection input
Next well input
Freeze input
Stop I/O
4 Programmable inputs
via RS232C
7.1.7 Options
Built-in Peltier cooling processing unit
Programmable Range: 4° C - 40° C
Cooling capacity:
Ambient -20° C
(measured on cooling plate, plate
holder removed )
Sample cooling
Column oven
User’s Manual FAMOS
Note: The temperature of the sample inside the sample vial may be
slightly different than the temperature set via the command, due to
a variety of effects (e.g. the heat transfer characteristics of the vial
walls). If it is necessary to precisely set the temperature inside the
vial, we recommend that you determine the temperature inside the
vial at various temperature setpoints and set the cooling option so
that the desired internal temperature is attained.
THERMOS column oven combined with the FAMOS Well
Plate Microautosampler. The injection valve and optional
high pressure valve fit into the column oven.
D931R1
7-3
Specifications
7.1.8 Plate Dimensions
FIGURE 7-1: Titerplate Formats
7-4
D931R1
User’s Manual FAMOS
Injection Principles
APPENDIX A
This appendix describes detailed information about the modes of injection used
by the FAMOS Well Plate Microautosampler.
A.1
Full Loop Injections
A.1.1 Standard Mode
The switching sequence for a full loop injection is described in this section.
At the start of the sequence, the injector is in the INJECT position (FIGURE A-1)
and the sample needle has entered the well after the air needle has pre-punctured
the septum. Headspace pressure is applied through the outer air needle to ensure
that no air or vapor bubbles are formed during sample aspiration.
FIGURE A-1. Injector is in the INJECT Position
The syringe dispenser aspirates the programmed flush volume from the sample
well to fill the sample line with sample and remove wash solvents (FIGURE A-2).
User’s Manual FAMOS
D931R1
A-1
Injection Principles
FIGURE A-2. Sample Line Filled with Sample
When the injection valve is switched into the LOAD position (FIGURE A-3), a
"sharp" sample front is placed at the inlet of the sample loop.
FIGURE A-3. Injection Valve Switched into LOAD Position
For full loop injections, the sample loop is quantitatively filled by transporting two
or more times the loop volume through the loop (FIGURE A-4). The actual volume
that is transported is dependent on the volume of the loop.
FIGURE A-4. Transporting Two (or more) Times the Loop Volume Through the Loop
The injection valve is then switched into the INJECT position (FIGURE A-5). The
sample loop is now part of the HPLC mobile phase flow path so that the sample
is transported to the column and the analysis time starts.
A-2
D931R1
User’s Manual FAMOS
Injection Principles
FIGURE A-5. The Injection Valve Switched into the INJECT Position
If one injection is to be done from each well or if a wash routine has to be
performed after every injection, the needle withdraws from the well directly after
the injection and, immediately washed (if programmed). After the analysis time is
completed, a new sequence is started.
If more than one injection is done from the same well without a wash routine,
the FAMOS Well Plate Microautosampler withdraws a flush volume after the
analysis time to compensate for diffusion of mobile phase from the rotor groove
into the first part of the sample line during the analysis time. The flush volume
between injections is not programmable and is always 50% of the programmed
flush volume. If the total amount of sample withdrawn with the next injection
from the well will exceed the total volume of the buffer tubing, the buffer tubing
is emptied into the wash position before the next injection. The next fill sequence
will then start with a full flush volume.
A.1.2 Using an Air Segment
An air segment can be used to reduce the amount of flush volume. This air
segment is at the front of the flush volume. It will not be injected and it will not
influence the injection. Use of an air segment can be enabled in the System
Menu (General Menu).
A = with air segment
B = without air segment
FIGURE A-6. Full Loop Injection with and without Air Segment
With the FAMOS Fused Silica needle, the flush volume must be at least 5.0 µL
for injections without air segment (if a stainless steel needle is used, the flush
volume should be at least 15.0 µL). If the samples are highly viscous, it may be
necessary to program larger flush volumes and reduce the syringe speed for
better performance.
User’s Manual FAMOS
D931R1
A-3
Injection Principles
A.2
Partial Loopfill Injections
A.2.1 Standard Mode
The switching sequence for a partial loopfill injection is schematically presented
in this section. The first three steps are identical to those for Full Loop injections
(see Section A.1).
For partial loopfill injections, the sample loop is filled by transporting the
programmed injection volume into the sample loop (FIGURE A-7).
FIGURE A-7. Partially Filling the Injection Loop with the Programmed Injection Volume
The injection valve switches into the INJECT position (FIGURE A-8) and the
sample loop is now part of the HPLC mobile phase flow path. At this point, the
sample is transported to the column and the analysis time starts.
FIGURE A-8. The Injection Valve Switched into the INJECT Position
The next injection sequence will start with a flush of 50% of the programmed
flush volume, if the next injection is from the same vial and no wash routine is
programmed. Otherwise it will start with a flush of the programmed flush
volume. If the aspiration of sample for the next injection will exceed the total
volume of the sample buffer tubing, the buffer tubing is emptied before the next
injection. The next injection will start with the programmed flush, as described
for full loop injections.
A-4
D931R1
User’s Manual FAMOS
Injection Principles
A.2.2 Using an Air Segment
An air segment can be used to reduce the amount of flush volume (FIGURE A-9).
This air segment is at the front of the flush volume and will not be injected. Use
of an air segment can be enabled in the System Menu (General Menu).
A = with air segment
B = without air segment
FIGURE A-9. Partial Loop Injection with and without Air Segment
User’s Manual FAMOS
D931R1
A-5
Injection Principles
A.3
µL Pick-Up Injections
A.3.1 Standard Mode
The switching sequence for a µL pick-up injection is schematically presented in
this section.
At the start of the sequence, the injection valve is in INJECT position (FIGURE A10). The sample needle has entered the vial of transport liquid (normally mobile
phase to avoid disturbance of the chromatogram with an additional peak of the
transport solvent) after the air needle has pre-punctured the septum. The
headspace pressure, applied through the outer air needle, ensures that no air or
vapor bubbles are formed during wash solvent aspiration.
FIGURE A-10. Injection Valve in Inject Position, Sample Needle in the Transport Vial
Note: When performing µL pick-up injections, use the mobile phase as the
transport liquid to avoid a disturbance of the chromatogram due to an additional
peak of the transport solvent).
For the first injection after a wash or after emptying of the buffer tubing, the
syringe dispenser aspirates transport liquid from the transport vial to fill the
sample line with transport liquid and remove wash solvent (FIGURE A-11).
FIGURE A-11. The Syringe Dispenser Aspirating Transport Liquid to Fill Sample Line
The needle moves from the transport vial to the sample well (FIGURE A-12). The
injection valve is switched to the LOAD position.
A-6
D931R1
User’s Manual FAMOS
Injection Principles
FIGURE A-12. The Injection Valve is Switched into LOAD Position
The programmed injection volume is aspirated from the sample well (FIGURE A13).
FIGURE A-13. Aspirating the Programmed Injection Volume
The sample needle moves back to the transport vial (FIGURE A-14). The sample
is quantitatively transported into the loop, with transport liquid (mobile phase)
from the transport vial.
FIGURE A-14. Transporting Sample into the Injection Loop (Aspirating the Programmed Injection
Volume)
The injection valve is switched to INJECT (FIGURE A-15). The sample loop is
now part of the HPLC mobile phase flow path and sample is transported to the
column. The analysis time is started.
User’s Manual FAMOS
D931R1
A-7
Injection Principles
FIGURE A-15. The Injection Valve in the INJECT Position (Aspirating the Programmed Injection
Volume)
The next sequence will skip the first withdrawal of transport solvent, unless a
wash routine is performed or the FAMOS Well Plate Microautosampler has
emptied the buffer tubing to waste. In those cases the sequence is completely
repeated.
A.3.2 Using an Air Segment
If an air segment has been programmed, it appears at the front of the first plug
of transport liquid and at the front of every sample plug. Use of an air segment
can be enabled in the System Menu (General Menu).
Note: The air segment at the front of the sample plug is injected into the HPLC
system.
A = with air segment
B = without air segment
FIGURE A-16. µL Pick-up Injections with and without Air Segment
A-8
D931R1
User’s Manual FAMOS
Injection Principles
A.4
Low Dispersion Injection
In addition to the standard injection methods, the FAMOS Well Plate
Microautosampler allows the analyst to control the injection profile via the “Low
Dispersion” routine in the partial loopfill and the full loopfill injection modes.
The injection principle is used to improve the injection profile (and therefore
improve the reproducibility) by switching the injection valve back into load
position after a calculated time and to cut off the tailing part of the sample plug.
Two parameters are used to control the Low Dispersion mode:
Flow Rate: This value determines the flow rate at which the sample is flushed
out of the injection loop. This value corresponds to the flow rate of the LC
system. The valid range is 00.00 to 99.99 µL/min
Note: If the flow rate is set to 00.00 µL/min, a standard injection is performed.
Low Dispersion Factor (L.D. factor): This value determines the amount of sample
volume which is cut off by the back switching injection valve. A value of 1.0
corresponds to the volume of the sample plug. If the value is greater than (less
than) 1.0, the injection valve will switch back after the volume is greater than
(less than) the sample volume which has been transferred through the sample
loop. The valid range is 0.7 to 2.0. As an example, if the value is 1.2, the valve
will switch back after 120 % of the value of the sample plug has been
transferred. The injection profile for various values is shown in FIGURE A-17.
1.3
1.1 1.0 0.9
L.D. factor
FIGURE A-17. Low Dispersion Injection Profiles
The time the injection valve is switched back into the load position is calculated
by the equation A-1.
St =(f) (V1) (60)/F
where:
A-1
St = Switching Time (sec), f = L.D. Factor,
V1 = Injection Volume (µL), F = Flow Rate (µL/min)
As an example: If Vinj = 1 µL, the flow rate is 5 µL and the L.D. factor is 1.1,
the valve switching time St will be 13.2 s.
Note: Under these conditions the loop will be flushed with the wash solvent as
well, which may result in injection problems (e.g. lack of proper focussing). It is
recommended that the mobile phase (typically mobile phase A) is used as the
wash solvent.
Note: The injection time must be at least 10 s. If the calculated injection time is
less than this limit, it will be redefined to 10 s.
User’s Manual FAMOS
D931R1
A-9
Injection Principles
A.5
Comparing the Various Sample Injection Modes
The characteristics of each injection mode is described below:
• Full loop injection - provides maximum reproducibility (RSD < 0.4%), but not
maximum accuracy, since loop volume is specified with an accuracy of ±
10%. Minimum sample loss = 12.4 µL (2 x loop overfill + flush volume for
needle) for the standard 5 µL loop.
• Partial loopfill - provides maximum accuracy (depends on syringe accuracy)
and reproducibility better than 0.6% RSD for injection volumes > 1 µL.
Minimum sample loss (Flush volume) = 5.0 µL. 5.0 µL is the recommended
minimum flush volume; smaller flush volumes can be programmed, but will
result in decreased performance. For maximum reproducibility and accuracy a
25 µL syringe should be used and a 1 µL sample loop should be used to avoid
loss of accuracy due to expansion of the loop content when switching from
inject to load position prior to sample loading. When working with high
pressure (200 bar), this loss may be up to 0.025 µL for a 5 µL loop.
Note: The minimum sample loss in partial loopfill mode is 5 µL (recommended
minimum flush volume) for the first injection and an additional 2.5 µL (always
half the programmed flush volume) for additional injections from the same well. If
a wash between injections has been programmed, sample loss is 5 µL for every
injection. For zero sample loss injections, use the µL-pick injection mode.
• µL Pick-up - means zero sample loss, maximum accuracy (same as partial
loopfill), but slightly diminished reproducibility: RSD better than 1.5% for
injection volumes ≥ 1 µL (sample volume must be larger then 10 µL). For
optimum accuracy and reproducibility, a 25 µL syringe and a 10 µL sample
loop should be used. The sample plug is transported into the loop, preceded by
a plug of transport liquid that has a volume of 2.5 times the programmed
needle tubing volume.
A-10
D931R1
User’s Manual FAMOS
Programming Charts
APPENDIX B
B.1
Overview of the Operating Program
The details of the operating program for the FAMOS Well Plate Microautosampler
is described in detail in Chapter 3, which includes a discussion of each
parameter, the allowable values and its role in the overall operation of the
autosampler.
The tables in this appendix are provided to allow the user to get an overview of
the various commands in each menu and how the various commands are
accessed.
The FAMOS Microautosampler
D931R1
B-1
Programming Charts
Ready Menu
Å [Menu] Æ
<PLATES>
<WASH>
<SYR END>
<UTILS>
<COPY
Exchange
Lift_left ***
Lift_right ***
Plate
[System]
Perform
standard wash
Move syringe
<SERIAL>
<ERASE>
<LOG>
<SSV> **
<COOL> **
<SERVICE>
• Cool ON/OFF
• Setpoint
• Switch ON/OFF
<DEFAULT ALL>
Copy method Erase methods Logbook
All methods
Communication Prime solvent
& Userprog
erased, all settings
protocol
valve
to default
Service mode
System Settings autosampler
<MICRO>
<CONVENTIONAL>
SystemSettings
<GENERAL>
<USAGE>
• Loop volume
• Needle tubing
• Syringe volume 1)
• Syringe speed
• Needle height
• Skip missing wells
• Air segment
• Headspace pressure
• Time base display
• Key click
• Error buzzer
• Alarm buzzer
1) only in Conventional mode
•
•
•
•
•
•
•
<PLATES>
Method protection code
Use time based method
Use mix method
Use user-program
Use labeled wells
Use templates
Use calibration wells
• Type of plate
• Process in
ROWS/COLUMNS
• Number of transport
vial
<IO>
•
•
•
•
•
•
•
Å [Menu] Æ
Inj. Marker pulse
Well marker pulse
Lab. Well marker pulse
Input edge next inj.
Input edge next well
Freeze input active
Reset outputs after
series
<CLOCK>
• System clock
• Date
• Time
<COMM.>
• Device identifier
Methods
[Method]
<INJECTION>
<WASH>
Injection methods
<FULL>
•
•
•
•
Analysis time
Flush volume
Injections/well
Inject. control:
STD/L.D.
• Flowrate
• L.D. -factor
[Series]
•
•
•
•
•
•
•
•
•
[Start/Stop]
[Priority]
[Interrupt]
[Help]
B-2
•
•
•
•
•
Timebase program:
<PARTIAL>
<PICK-UP>
Analysis time
Flush volume
Injections/well
Inj. Volumes
Inject. control:
STD/L.D.
• Analysis time
• Injection/well
• Inj. volumes
• Wash
between:
– Series
– Wells
– Injections
Injection method
Wash method
Time base method
Well number
•
•
•
•
•
•
•
•
4x AUX 1
4x AUX 2
4x AUX 3
4x AUX 4
4x ISS-A
4 x ISS-B
8x CODE OUT
END TIME
• Flowrate
• L.D. -factor
Mix method
Injection method
Wash method
Time base method
Use calibration wells (yes/no)
– calibration wells
– calibration interval
Sample wells
Destination wells
Reagent wells
Labeled wells
<MIX> *
• Wash volume
Mix method:
• ASPIRATE
– sample
– destination
– air
– reagent
• DISPENSE
– sample
– destination
– waste
– reagent
• REPEAT
• WAIT
• WASH
*
*
*
When the conventional autosampler set up is selected in
the system setting, the value ranges of the system settings
and in the methods will be adjusted to the needs of the
conventional autosampler.
Also will the Low Dispersion mode in the injection methods
no longer be available
*
*
*
• First series
• Last series
• <START> <REMOTE>
•
•
•
•
<TIMEBASE> *
Note: *
Note: **
Note: ***
*
Not used
These parameters depend on the contents of
the methods used and/or the System
Settings of the FAMOS
optional Soft-function keys:
only if Feeder option is installed
PRIORITY:
Only available when user-program and mix method have
been disabled in the System Settings.
Context and status sensitive help screen (if available)
D931R1
User’s Manual FAMOS
Programming Charts
Ready Menu
<EXCHANGE>
<.PLATES>***
Å [Menu] Æ
<WASH>
<SYR END>
<UTILS>
<COPY
Exchange
Lift_left
Lift_right
Plate
[System]
Perform
standard wash
Move syringe
<SERIAL>
<ERASE>
<LOG>
<COOL> **
•
•
•
<DEFAULT ALL>
<SERVICE>
Cool ON/OFF
Setpoint
Switch
ON/OFF
Copy method Erase methods Logbook
All methods
Communication
& Userprog
erased, all settings
protocol
to default
Service mode
System Settings autosampler
<MICRO>
<CONVENTIONAL>
System Settings
<GENERAL>
<USAGE>
• Loop volume
• Needle tubing
• Syringe volume 1)
• Syringe speed
• Needle height
• Skip missing wells
• Air segment
• Headspace pressure
• Time base display
• Key click
• Error beep
• Alarm buzzer
1) only in Conventional mode
•
•
•
•
•
•
•
<PLATE>
Method protection code
Use time base method
Use mix method
Use user-program
Use labeled wells
Use templates
Use calibration wells
• Type of plate
• Process in
ROWS/COLUMNS
• Number of transport
vial
<IO>
•
•
•
•
•
•
•
Å [Menu] Æ
<CLOCK>
• System clock
• Date
• Time
Inj. Marker pulse
Well marker pulse
Lab. Well marker pulse
Input edge next inj.
Input edge next well
Freeze input active
Reset outputs after
series
<COMM.>
• Device identifier
Methods programming
[Method]
<TEMPLATE>
<METHODE>
Methods
• Userprog
*
or
• Mix method
*
• Injection method
• Wash method
• Timebase meth.*
<FLUSHED>
Analysis time
Flush volume
Injections/well
Inject. control:
STD/L.D.
• Flowrate
• Delay factor
[Priority]
With templates
• Template
• Use calibration wells (yes/no) *
– calibration wells
– calibration interval
• Sample wells
• Destination wells
*
• Reagent wells
*
• Labeled wells
*
•
•
•
•
•
<PARTIAL>
<PICK-UP>
Analysis time
Flush volume
Injections/well
Inj. volumes
Inject. control:
STD/L.D.
• Analysis time
• Injection/well
• Inj. Volumes
<TIMEBASE> *
• Wash volume
• Wash between:
– Series
– Wells
– Injections
•
•
•
•
•
•
•
•
Without templates
• Userprogram (yes/no)
or
• Mix method
• Injection method
• Wash method
• Time base method
• Use calibration wells (yes/no)
– calibration wells
– calibration interval
• Sample wells
• Destination wells
• Reagent wells
• Labeled wells
Mix method
4x AUX 1
4x AUX 2
4x AUX 3
4x AUX 4
4x ISS-A
4x ISS-B
8x CODE OUT
END TIME
• ASPIRATE
– sample
– destination
– air
– reagent
• DISPENSE
– sample
– destination
– waste
– reagent
• REPEAT
• WAIT
• WASH
•
•
•
•
Injection method
Wash method
Timebase method
Vial number
•
•
•
•
•
•
•
•
•
•
•
•
•
ASPIRATE
DISPENSE
SYR-VALVE
SYRINGE
WASH
VALVE
WAIT
COMPRESS
AUX
WAIT INPUT
PROG-OUT
CODE
MARKERS
*
*
*
*
*
*
*
When the conventional autosampler set up is selected in the
system setting, the value ranges of the system settings and
in the methods will be adjusted to the needs of the
conventional autosampler.
Also will the Low Dispersion mode in the injection methods
no longer be available
Note: *
Note: **
Note: ***
• Template
• Vial number
<MIX> *
Timebase program
• Flowrate
• Delay factor
• First series
• Last series
• <START> <REMOTE>
[Interrupt]
[Help]
<WASH>
Injection methods
•
•
•
•
[Start/Stop]
User program
<INJECTION>
2
[Series}
<USRPROG> *
*
These parameters depend on the contents of
the methods used and/or the System Settings
of the FAMOS
optional Soft-function keys:
only if Feeder option is installed
PRIORITY:
Only available when user-program and mix method have been
disabled in the System Settings.
Not used
Context and status sensitive help screen (if available)
User’s Manual FAMOS
D931R1
B-3
Programming Charts
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B-4
D931R1
User’s Manual FAMOS
Additional Programming
Examples
APPENDIX C
C.1
Overview
The commands described in Chapter 3 can be used by the analyst to generate a
broad range of programs for the operation of the FAMOS Well Plate
Microautosampler. These commands allow the user to configure the unit to meet
the specific needs of the laboratory. A series of blank forms are provided to
assist the analyst in programming (Appendix F).
Chapter 4 describes a number of programs that the user can prepare for activities
that are commonly performed with the microautosampler. In this appendix, we
describe a few programs that are important, but used less frequently. In addition
to the direct use of these programs, they can be used to provide an
understanding of the logical processes that are used in generating a program.
The following programs are included in this appendix.
•
A 1.0 µL partial loopfill (Section C.2)
•
A 3 x 1.0 µL injection with µL pick-up partial loopfill injection (Section C.3)
•
A 1:10 dilution followed by a 1.0 µL partial loopfill injection (Section C.4)
•
Defining a Template and adding a protection code (Section C.5)
•
1 µL injections out of 25 wells using a 384 well plate (Section C.6)
•
Column Switching (Section C.7)
It should be noted that the examples described below could be used with
systems that include UltiMate with the UltiChrom software program as well as
for stand-alone systems.
User’s Manual FAMOS
D931R1
C-1
Additional Programming Examples
C.2
A 1.0 µL Partial Loopfill
This example describes an application in which a 1.0 µL injection is made from a
single vial in position 1. In this example, we assume that a 5 µL loop, a 2.4 µm
needle and a 25 µL syringe have been installed.
a) Place a sample in position A1.
b) Program the System Parameters by entering the parameters indicated in
TABLE C-1.
TABLE C-1. Programming System Parameters
Press keys
System
<MICRO> E
<GENERAL> E
[05.0] E
[02.4] E
<LOW> E
[02] E
<NO> E
<YES> E
Escape
<PLATES> E
<96-LOW>
<IN ROWS>
Escape Escape
to
to
to
to
to
to
to
to
to
to
to
to
to
to
Description
enter the System Menu
select the micro mode
enter the General Menu
define the volume of the installed loop
define the volume of the needle tubing
set syringe speed to low
set sample needle height to 2 mm
enable use of air segment
switch on headspace pressure
return to the System Menu
enter the Plates Menu
define the type of plate to be used
define the processing order of wells
return to the Ready Menu
For this example, all other settings used will be the default value. In the tables
presented in this appendix the name of the soft key to be selected is indicated in
angle brackets (e.g. <PLATES>), and the value which is indicated in square
brackets (e.g. [05.0]). The bold letters indicates that you have to press a
function key (e.g. E which represents Enter).
c) Program the method by entering the parameters indicated in TABLE C-2.
TABLE C-2. Programming Method Parameters
Press keys
Methods
<INJECTION> [01] E
<PARTIAL> E
[100] E
[05.0] E
[1] E
[1.00] E
Escape Escape
C-2
Description
to enter the Methods Menu
program injection method number 1
to select partial loopfill injection method
to define an analysis time of 1 minute
to define a flush volume of 5.0 µL
to define the number of injections per well
to set the injection volume at 1.00 µL
to return to the Ready Menu
D931R1
User’s Manual FAMOS
Additional Programming Examples
d) Program the series by entering the parameters indicated in TABLE C-3.
TABLE C-3. Programming Series Parameters
Press keys
Series
[01] E
[01] E
CL E
<ROW A> [01] E
<ROW A> [01] E
Escape
Description
to enter the Series Menu
to define the Series number
to define the injection method number
to enter <NONE> for wash method
to define location of the first sample well
to define location of the last sample well
to return to the Ready Menu
e) Run the series by entering the parameters indicated in TABLE C-4 and
pressing <START>.
TABLE C-4. Running the Series
Press keys
Start/Stop
[01] E
[01] E
<START>
to
to
to
to
Description
start the FAMOS Well Plate Microautosampler
start at series number 1
stop after execution of series number 1
start the analytical run
The FAMOS Well Plate Microautosampler will now locate well A01 and perform a
1.0 µL partial loopfill injection. The display of the FAMOS Well Plate
Microautosampler will indicate the status (Checking tray, Flushing, Loopfill,
Running, Rinse buffer, Running). The display also indicates the number of the
defined series (01), the method number (01) and the well on which the analysis
is performed (A01).
At the end of the defined analysis time, the Ready Menu will be displayed again
to indicate that the microautosampler is ready for the next analytical run.
User’s Manual FAMOS
D931R1
C-3
Additional Programming Examples
C.3
A 3 x 1.0 µL Injection with µL Pick-Up and Wash between
Injections
This example describes an application where three 1.0 µL injections are made,
with a wash between each injection.
a) Place a vial with transport solvent (mobile phase) in transport vial position 1
(left) and place the sample in position A1. Make sure the transport vial is
correctly filled before starting a new series.
b) Program the System Parameters by entering the parameters indicated in
TABLE C-5.
TABLE C-5. Programming System Parameters
Press keys
System
<MICRO> E
<GENERAL> E
E until ‘Air segment’
appears
<NO> E
Escape
<PLATES> E
EE
[01] E
[01]
Escape Escape
to
to
to
to
Description
enter the System Menu
select the micro mode
enter the General Menu
go to the Air segment field
to
to
to
to
to
to
to
switch off air segment
return to the System Menu
enter the Plates Menu
go to transport vials field
define position of the first transport vial
define position of the last transport vial
return to the Ready Menu
e) Program the method by entering the parameters indicated in TABLE C-6.
TABLE C-6. Programming Method Parameters
Press keys
Methods
<INJECTION> [02] E
<PICK-UP> E
[100] E
[3] E
[100] E
[100] E
[100] E
Escape
<WASH>
[01] E
<INJECTION> E
[50]
Escape Escape
to
to
to
to
to
to
to
to
to
to
to
to
to
to
Description
enter the Methods Menu
define method number 02
select the injection mode for this method
define the analysis time
define the number of injections per well
define volume of 1.0 µL for 1st injection
define volume of 1.0 µL for 2nd injection
define volume of 1.0 µL for 3rd injection
return to the Methods Menu
enter the Wash Menu
define wash method number 01
select wash between injections
define the wash volume
return to the Ready Menu
Note: Make certain to indicate that a 10 µl loop is used in the system settings
when µL pick-up injections are selected (Section 3.8.2).
C-4
D931R1
User’s Manual FAMOS
Additional Programming Examples
f)
Program the series by entering the parameters indicated in TABLE C-7.
TABLE C-7. Programming Series Parameters
Press keys
Series
[01] E
[02] E
[01] E
<ROW A> [01] E
<ROW A> [01]
Escape
to
to
to
to
to
to
to
Description
enter the Series Menu
define the series number
define the injection method for this series
define the wash method for this series
define the location of the first sample well
define the location of the last sample well
return to Ready Menu
g) Run the series by entering the parameters indicated in TABLE C-8 and
pressing <START>.
TABLE C-8. Running the Series
Press keys
Start/Stop
[01] E
[01] E
<START>
to
to
to
to
Description
start the FAMOS Well Plate Microautosampler
start at series 01
stop after series 01
start execution of the series.
At the end of the defined analysis time the Ready Menu will appear again to
indicate that the FAMOS Well Plate Microautosampler is ready for the following
next run.
User’s Manual FAMOS
D931R1
C-5
Additional Programming Examples
C.4
A 1:10 Dilution followed by a 1.0 µL Partial Loopfill Injection
This example describes how to let the FAMOS Well Plate Microautosampler
transfer 9 µL from Reagent A to the destination vial, add 1 µL of sample,
followed by subsequently inject 1.0 µL (3 times). In addition, a 96 deep well
plate is used and some wait steps are included to increase the performance.
a) Place a sample vial in position A 01, place an empty sample vial in position
B 01 as destination vial. Place a filled reagent vial in position 1. Make sure
the reagent vial is filled correctly before starting a new series.
b) Program the System Parameters by entering the parameters indicated in
TABLE C-9.
TABLE C-9. Programming System Parameters
Press keys
System
<CONVENTIONAL> E
<USAGE> E
EE
<ENABLED>
Escape
<PLATES> E
<96-HIGH>
<COLUMNS>
CL
Escape Escape
to
to
to
to
to
to
to
to
to
to
to
Description
enter the System Menu
select the conventional mode
enter the Usage Menu
go to the Mix field
enable use of mix methods
return to the System Menu
enter the Plates Menu
select 96 deep well plate
select processing in columns (A1, B1, etc)
disable use of transport vials
return to the Ready Menu
Note: As soon as a change has been entered in the System settings, the
message “ALL SERIES DEFAULT” appears. The user will have to redefine series
because the settings have been changed.
c) Program the method by entering the parameters indicated in TABLE C-10.
TABLE C-10. Programming Methods Parameters
Press keys
Methods
<INJECTION> [03] E
<PARTIAL> E
[100] E
[50] E
[3] E
[100] E
[100] E
[100] E
Escape
to
to
to
to
to
to
to
to
to
to
Description
enter the Methods Menu
enter the Injection Menu
select partial loopfill injection mode
define the analysis time
define the flush volume
define the number of injections per well
enter the injection volume for 1st injection
enter the injection volume for 2nd injection
enter the injection volume for 3rd injection
return to the Methods Menu
d) Program the mix method by entering the parameters indicated in TABLE C11.
C-6
D931R1
User’s Manual FAMOS
Additional Programming Examples
TABLE C-11. Programming Mix Methods Parameters
Press keys
<MIX> [1] E
<INSERT>
<ASPIRATE> [100] E
Menu <REAG-A> E
<INSERT>
<DISPENSE> [90]
<DESTINATION>
<INSERT>
<WAIT> [02] E
<INSERT>
<DISPENSE> [10]
<WASTE>
<INSERT>
<WAIT> [02] E
<INSERT>
<ASPIRATE> [20] E
<AIR>
<INSERT>
<WAIT> [02] E
<INSERT>
<ASPIRATE> [20] E
<SAMPLE>
<INSERT>
<WAIT> [02] E
<INSERT>
<DISPENSE> [10] E
<DESTINATION>
<INSERT>
<WAIT> [02] E
<INSERT>
<DISPENSE> [30] E
<WASTE>
<INSERT> <MENU>
WASH [300] E
<INSERT>
<ASPIRATE> [20] E
<AIR>
<INSERT>
<ASPIRATE> [50]
<DESTINATION>
<INSERT>
<DISPENSE> [50]
<DESTINATION>
<INSERT>
<REPEAT> [2] 3[2] E
<INSERT>
<DISPENSE> [20] E
<WASTE>
<INSERT> <MENU>
<WASH> [300] E
User’s Manual FAMOS
Description
to enter the Mix Menu and define Mix method no. 1
to define mix method step number 1
to aspirate 10.0 µL from reagent vial A
to define mix method step number 2
to dispense 9.0 µL to destination well
to
to
to
to
define mix method step number 3
wait 2s
define mix method step number 4
dispense 1.0 µL to waste
to
to
to
to
define mix method step number 5
wait 2s
define mix method step number 6
aspirate 2.0 µL air
to
to
to
to
define mix method step number 7
wait 2s
define mix method step number 8
aspirate 2.0 µL sample
to
to
to
to
define mix method step number 9
wait 2s
define mix method step number 10
dispense 1.0 µL to the destination well
to
to
to
to
define mix method step number 11
wait 2s
define mix method step number 12
dispense 3.0 µL to waste
to
to
to
to
define mix method step number 13
perform a wash with 300 µL
define mix method step number 14
aspirate 2.0 µL air
to define mix method step number 15
to aspirate 5.0 µL from the destination well
(note: step will improve
to define mix method step number 16
to dispense 5.0 µL to the destination well
(step 15 and 16 are used to mix the destination)
to define mix method step number 17
to repeat last 2 steps 2 times
to define mix method step number 18
to dispense 2.0 µL to waste
to define mix method step number 19
to perform a wash with 300 µL
D931R1
C-7
Additional Programming Examples
e) Program the series by entering the parameters indicated in TABLE C-12.
TABLE C-12. Programming Series Parameters
Press keys
Series
[01] E
[01] E
[03] E
CL E
<ROW A> [1] E
<ROW A> [1] E
<ROW B> [1] E
[1] E
Escape
f)
to
to
to
to
to
to
to
to
to
to
Description
enter the Series Menu
define series number 1
select Mix method number 1 for this series
select Injection method number 3
select <NONE> for wash method
define location of first sample well
define location of last sample well
define location of first destination well
define position 1 for the Reagent A vial
return to the Ready Menu
Run the series by entering the parameters indicated in TABLE C-13 and
pressing <START>.
TABLE C-13. Programming System Parameters
Press keys
Start/Stop
[01] E
[01] E
<START>
to
to
to
to
Description
start the FAMOS Well Plate
start at series number 1
stop after series number 1
start processing of sample
The FAMOS Well Plate will now start searching for the Reagent vial and
transport 90 µL to the destination well once, then 1 µL of sample will be added
and after mixing 3 times a 1 µL injection will be performed.
C-8
D931R1
User’s Manual FAMOS
Additional Programming Examples
C.5
Defining a Template and Adding a Protection Code
This example describes how to incorporate an injection method (02) and a wash
method (01) which were defined in Section C.3 into a template. A protection
code will be added.
After use of templates has been enabled, the message “ALL SERIES DEFAULT”
appears. The user will have to redefine the series because the settings have been
changed.
a) To select the methods to be incorporated in the template, enter the
parameters described in TABLE C-14:
TABLE C-14. Programming the Method
Press keys
Methods
[123456] E
<TEMPLATE>
[01] E
[02] E
[01]
Escape Escape
Description
to enter the Methods Menu
to enter the methods protection code
to enter the Template Menu
to define the number for the template
to define the injection method for this
template
to define the wash method for this
template
to return to the Ready Menu
b) Program the series by entering the parameters indicated in TABLE C-15.
TABLE C-15. Programming Series Parameters
Press keys
Series
[01] E
[01] E
<ROW A> [01] E
<ROW B> [01] E
Escape
to
to
to
to
to
to
Description
enter the Series Menu
define the Series number
define the Template method number
define the first sample well
define the last sample well
return to the Ready Menu
c) Run the series by entering the parameters indicated in TABLE C-16 and
pressing <START>.
TABLE C-16. Running the Series
Press keys
Start/Stop
[01] E
[01] E
<START>
to
to
to
to
Description
start the Microautosampler
start analysis at series 01
stop after analysis of series 01
start the analytical run
The FAMOS Well Plate Microautosampler now performs the same actions as in
Example 2, except that analysis is performed on two wells: A 01 and B 01.
Note: Select <DEFAULT ALL> in the Ready Menu (Utilities Menu) to erase all
series and methods defined in these examples and to default all settings.
User’s Manual FAMOS
D931R1
C-9
Additional Programming Examples
C.6
1µ
µL injections out of 25 wells of a 384 well plate
This example describes the injection of 1 µL from out of 25 wells from a 384
well plate.
a) Place the samples in well positions B5-B14, G14-G22 and M3-M8.
b) Enter the systems settings command from the <GENERAL> menu and
select the 384 well plate.
c) Program the injection method as shown in TABLE C-17.
TABLE C-17. Programming the Injection Method
Press keys
Methods
<INJECTION> [01] E
<PARTIAL> E
[25.00] E
[05.0] E
[1] E
[1.00] E
<STD> E
Escape Escape
Description
to enter the Methods Menu
program injection method number 1
to select partial loopfill injection method
to define an analysis time of 25 minutes
to define a flush volume of 5.0 µL
to define the number of injections/vial
to set the injection volume to 1.00 µL
to select the standard injection control
to return to the Ready Menu
d) Program the three series as shown in TABLE C-18.
TABLE C-18. Programming Series Parameters
Press keys
Series
[01] E
[01] E
CL E
<ROW B> [05] E
<ROW B> [14] E
Series
[02] E
[02] E
CL E
<ROW G> [14] E
<ROW G> [22] E
Series
[03] E
[03] E
CL E
<ROW M> [03] E
<ROW M> [08] E
Escape
C-10
to
to
to
to
to
to
to
to
to
to
to
to
to
to
to
to
to
to
to
Description
enter the Series Menu
define the Series number
select the injection method number
enter <NONE> for wash method
define location B5 as first sample vial
define location B14 as last sample vial
enter the Series Menu
define the next Series number
select the next injection method number
enter <NONE> for wash method
define location G14 as first sample vial
define location G22 as last sample vial
enter the Series Menu
define the next Series number
select the next injection method number
enter <NONE> for wash method
define location M3 as first sample vial
define location M8 as last sample vial
return to the Ready Menu
D931R1
User’s Manual FAMOS
Additional Programming Examples
e) Run the series by entering the parameters indicated in TABLE C-19 and
pressing <START>.
TABLE C-19. Running the Series
Press keys
Start/Stop
[01] E
[03] E
<START>
to
to
to
to
Description
start the FAMOS Well Plate Microautosampler
start at series 01
stop after series 01
start execution of the series.
At the end of the defined analysis time, the Ready Menu will reappear to indicate
that the FAMOS Well Plate Microautosampler is ready.
User’s Manual FAMOS
D931R1
C-11
Additional Programming Examples
C.7
Column Switching
The FAMOS Well Plate Microautosampler is also available with an additional ISS
valve (Integrated Stream Switch option) which can be used for column switching
experiments. In this section we provide an example of the use of a pre-column
prior to the capillary HPLC analysis.
The following equipment is needed to perform this column switching example:
•
A FAMOS Well Plate Microautosampler with:
- the ISS-A valve (ISS in position A)
- a 1000 µL syringe (cat. no. FMS-SY-1000)
- a 2000 µL buffer tubing (cat. no. FMS-BT-2000)
- a 250 µL injection loop
- a standard injection needle assembly (cat. no. FMS-NDL-CNV) installed
•
An additional HPLC pump for loading the sample at a flow rate of
0.5 mL/min.
•
A µ-precolumn holder (cat. no. HD-05) and precolumns, e.g. 500 µm I.D. x 5
mm, 5 µm C18 (cat. no. MCA-50-C18).
To prepare the system:
•
Replace the injection needle assembly (Section 6.3.3).
•
Replace the 25 µL syringe for the 1000 µL syringe (Section 6.3.1) and
replace the buffer tubing.
•
Connect the contact closure input of the data system to the AUX 1 contact
of the autosampler (P5 connector) in order to start the data acquisition at the
moment the pre-column is switched on-line.
•
Set the system up according to FIGURE C-1.
FIGURE C-1. Column Switching Setup
C-12
D931R1
User’s Manual FAMOS
Additional Programming Examples
•
Set the FAMOS Well Plate Microautosampler in <CONVENTIONAL> mode
and adjust the general system parameters according to the modified setup
(e.g. syringe, loop volume (Section 4.2).
•
Enable the use of “time based methods” to allow programming of the AUX 1
contact closure (Section 3.8.4 and 3.8.12).
•
Program “injection method 01” with the following parameters:
”full loop” injection
”flush volume”: 30 µL
”number of injections per vial”: 1
•
Program “Time Base 01” with the parameters shown in TABLE C-20.
TABLE C-20. Programming Time Base
Parameter
Time
ISS-A 1-6
00:45
ISS-A 1-2
14:50
AUX-1
AUX-1
00:45
00:46
Comment
switches pre-column on-line after sampler
loading
switches the ISS-A valve back and the
pre-column off-line again for the next
sample
starts data acquisition of the data system
resets the contact closure output
•
Program “Series 01” using “Injection Method 01” and “Time Base Method
01”
•
Start the FAMOS Well Plate Microautosampler.
Note: The above example shows how to perform column switching with the
FAMOS Well Plate Microautosampler equipped with one ISS valve. For advanced
column switching applications (e.g. on-line digestion of proteins, on-line removal
of non-ionic detergents from protein/peptide samples) LC Packings offers the
SWITCHOS I (equipped with two valves) and the Switchos II (equipped
additionally with a Loading Pump and Solvent Selection Valve) Advanced Micro
Column Switching Units. Please contact LC Packings for additional information.
User’s Manual FAMOS
D931R1
C-13
Additional Programming Examples
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C-14
D931R1
User’s Manual FAMOS
The Sample Injector
APPENDIX D
D.1
Overview
The Cheminert Model C2 Injection Valve is a 6-port external loop injector
manufactured by Valco Instruments, Co. Inc. It is designed to offer optimal
results with any of the injection principles used in the FAMOS Well Plate
Microautosampler, including the partial filling and the µL Pick-Up methods (in
which the injection volume is determined by the syringe) and full loop injection
(in which the volume is determined by the size of the loop). The design prevents
any contact between the needle and the stator and rotor faces.
Note: A detailed discussion on the Installation, Use and Maintenance of the valve
is presented in Technical Note 801 from Valco Instruments, Co. Inc. and can be
obtained at the Valco website (www.Valco.com)
User’s Manual FAMOS
D931R1
D-1
The Sample Injector
D.2
Maintenance
In most instances, the only maintenance that is required is cleaning of the valve.
Cleaning can often be accomplished by flushing all the lines with an appropriate
solvent(s). The selection of the solvent is dependent on the nature of the sample
and the mobile phases that are used. Typically solvents such as methanol,
acetonitrile, methanol/water (80/20) or acetonitrile/water (80/20) should be
used.
D.3
Disassembly/Reassembly of the Valve
Note: Do not disassemble the valve unless system malfunction is definitely
isolated to the valve.
D.3.1 Disassembly of the Valve
To disassemble the valve:
a) Use a 9/16” hex driver to remove the socket head screws which secure the
cap to the valve (FIGURE D-1)
Driver
Rotor
Stator
FIGURE D-1. Exploded View of the Valco Model C2 Valve
b) To insure that the sealing surface of the cap is not damaged, rest it on its
outer face. If the tubing is still attached, leave it suspended by the tubing.
c) Gently pry the rotor away from the driver with your fingers or a small
screwdriver.
d) Examine the rotor sealing surface for scratches.
•
If scratches are visible to the naked eye, the rotor must be replaced.
•
If no scratches are visible, clean all parts thoroughly with an
appropriate solvent. Take care that no surfaces get scratched while
you are cleaning the components.
Note: The most common problem in the use of the valve with HPLC is the
formation of buffer crystals, which are usually water soluble. After cleaning, it is
not necessary to dry the rotor.
D-2
D931R1
User’s Manual FAMOS
The Sample Injector
D.3.2 Reassembly of the Valve
To reassemble the valve:
a) Replace the rotor in the driver, making sure that the rotor sealing surface
with its engraved flow passages is facing out. The pattern is asymmetrical to
prevent improper placement.
b) Replace the cap. Insert the two socket head screws and tighten them gently
until both are snug.
CAUTION
Caution: Do not overtighten the screws-they simply hold the assembly together
and do not affect the sealing force, which is automatically set as the screws
close the cap against the valve body.
c) Test the valve by pressurizing the system. If the valve does not hold pressure
it should be returned for repair.
Note: When re-installing the valve, make certain that the proper tubing is
attached to the appropriate fitting. The loop is connected to ports 1 and 4, the
pump is connected at port 2 and the column is connected at port 3.
User’s Manual FAMOS
D931R1
D-3
The Sample Injector
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D-4
D931R1
User’s Manual FAMOS
Digital Inputs and Outputs
APPENDIX E
E.1
Introduction
The rear panel of the FAMOS Well Plate Microautosampler includes a series of
I/O connectors that allow the user to interface the microautosampler to other
components in their analytical system.
If the microautosampler is used with a LC Packings UltiMate Capillary and Nano
HPLC systems, refer to the User’s Manual for that system for specific
installation/interfacing information.
User’s Manual FAMOS
D931R1
E-1
Digital Inputs and Outputs
E.2
Contact Closure Outputs
I/O Connector P1 (programmable outputs), P4 (marker outputs) and P5 (auxiliary
outputs) are contact closure outputs (floating NO/NC contact) as shown in
FIGURE E-1.
Normally Closed
(NC)
Common
(COM)
Normally Open
(NO)
FIGURE E-1. Contact Closure Output
E.2.1 P1 Connector – OUTPUTS
TABLE E-1. P1 Connector Pin Descriptions
PIN #
1
2
3
4
5
6
13
Name
OUT 1
OUT 2
Function
Normally open
Common
Normally closed
Normally open
Common
Normally closed
24 V DC
Normally Closed
(NC)
Common
(COM)
Normally Open
(NO)
PIN #
7
8
9
10
11
12
14
15
Name
(Spare)
Alarm
output
Function
Normally open
Common
Normally closed
Normally open
Common
Normally closed
Power ground
Note:
VMAX = 28 VDC / VAC, IMAX = 0.25 A
Connector P1 OUTPUTS includes 2 programmable outputs and an alarm output
Note: The Alarm output will be activated whenever an error occurs. A description
of the error codes of the FAMOS Well Plate Microautosampler is presented in
Chapter 6.
E.2.2 P4 Connector – MARKERS
TABLE E-2. P4 Connector Pin Descriptions
PIN #
1
2
3
4
5
6
13
Name
Inject
Marker
Well
Marker
Function
Normally open
Common
Normally closed
Normally open
Common
Normally closed
24 V DC
Common
(COM)
Normally Closed
(NC)
Normally Open
(NO)
PIN #
7
8
9
10
11
12
14
15
Name
Labeled
Well
Marker
Stop I/O
Function
Normally open
Common
Normally closed
Normally open
Common
Normally closed
Power ground
Note:
VMAX = 28 VDC / VAC, IMAX = 0.25 A
VMAX =28 VDC/VAC, IMAX = 0.25 A
E-2
D931R1
User’s Manual FAMOS
Digital Inputs and Outputs
E.2.3 P5 Connector – AUXILIARIES
TABLE E-3. P5 Connector Pin Descriptions
PIN #
1
2
3
4
5
6
13
Name
AUX 1
AUX 2
Function
Normally open
Common
Normally closed
Normally open
Common
Normally closed
24 V DC
Common
(COM)
Normally Closed
(NC)
Normally Open
(NO)
PIN #
7
8
9
10
11
12
14
15
Name
AUX 3
AUX 4
Function
Normally open
Common
Normally closed
Normally open
Common
Normally closed
Power ground
Note:
VMAX = 28 VDC / VAC, IMAX = 0.25 A
VMAX =28 VDC/VAC, IMAX = 0.25 A
User’s Manual FAMOS
D931R1
E-3
Digital Inputs and Outputs
E.3
TTL Outputs
The following tables show the marker outputs (P2) and a 4 bit time base code
output (P3), programmable in a time base method. Both connectors are TTL level
outputs (FIGURE E-2).
Autosampler
Ext. device
FIGURE E-2. TTL outputs
E.3.1 P2 Connector - TTL OUTPUTS
TABLE E-4. P2 Connector Pin Descriptions
PIN #
Function
INJECT MARKER
1
VIAL/WELL MARKER
2
LABELED WELL MARKER
3
STOP I/O
4
not connected
5
not connected
6
7
not connected
not
connected
8
PIN #
Function
not connected
9
not connected
10
not connected
11
not connected
12
Signal ground
13
Signal ground
14
15
Signal ground
All markers are active low (logical 0).
VMAX = 5.5 V, logical 1 > 3.5 V , logical 0 < 1.0 V.
DC output source / sink current ± 20 mA.
Note: A marker output pulse will be generated when the injection valve switches
from LOAD to INJECT. However, in a User Program markers have to be
programmed by the user.
E.3.2 P3 Connector - TIMED OUTPUTS
TABLE E-5. P3 Connector Pin Descriptions
Connector P3 TIMED OUTPUTS; 4 bit time base code output
PIN #
Function
TB 0 (HEX) (1)
1
TB 1 (HEX) (2)
2
TB 2 (HEX) (4)
3
TB 3 (HEX) (8)
4
not used
5
PIN #
Function
Signal ground
6
7
Signal ground
Signal ground
8
Signal ground
9
All markers are active low (logical 0).
VMAX = 5.5 V, logical 1 > 3.5 V , logical 0 < 1.0 V.
DC output source / sink current ± 20 mA.
E-4
D931R1
User’s Manual FAMOS
Digital Inputs and Outputs
E.4
TTL Inputs
The TTL input connector (FIGURE E-3) is an active high or active low TTL input;
it can be defined in the System Menu. The NEXT INJECTION INPUT and the
NEXT WELL INPUT can be used when the FAMOS Well Plate works in REMOTE
CONTROL. The FREEZE INPUT and STOP I/O input can be used to control the
FAMOS Well Plate by other devices. The four inputs (INPUT 1 to 4) can only be
used in the user program, e.g. to control the sequence of the steps in this
method. A connection diagram is shown in FIGURE E-3.
Autosa mpler
Autosa mpler
FIGURE E-3. TTL Input
E.4.1 P6 Connector - TTL Inputs
TABLE E-6. P6Connector Pin Descriptions
PIN #
Function
NEXT INJECTION INPUT
1
NEXT WELL INPUT
2
FREEZE INPUT
3
STOP I/O
4
INPUT 1
5
INPUT 2
6
INPUT 3
7
INPUT 4
8
PIN #
Function
Signal ground
9
Signal ground
10
Signal ground
11
Signal ground
12
Signal ground
13
Signal ground
14
Signal ground
15
Next injection input: This input will start the next injection sequence when the
FAMOS Well Plate Microautosampler is started in remote control. When the
injection sequence is finished the FAMOS Well Plate Microautosampler will wait
for the next input.
From the Ready Menu, a NEXT INJECTION INPUT will start the last programmed
series. In this case the FAMOS Well Plate Microautosampler will not wait for the
NEXT INJECTION INPUT before continuing with the next injection. The FAMOS
Well Plate Microautosampler will execute the complete RUN as if it was started
with the Start/Stop key.
Next well input: With this input the FAMOS Well Plate will perform the next
injection from the next well, even if not all injections from that well in the
programmed injection method have been executed.
User’s Manual FAMOS
D931R1
E-5
Digital Inputs and Outputs
Freeze input: The FAMOS Well Plate Microautosampler will freeze the analysis
time for the time that this input is active. If the FREEZE INPUT is activated while
the analysis time is not running, the FAMOS Well Plate Microautosampler will
perform all programmed pre-injection sample handling (mix method and loading
part of the injection method). It should be noted that the FAMOS Well Plate
Microautosampler will not inject the sample until the FREEZE INPUT is no longer
active.
Stop I/O: With this input the run of the FAMOS Well Plate Microautosampler is
immediately aborted and the Ready Menu appears in the display. If the FAMOS
Well Plate Microautosampler is in remote control, the run of the FAMOS Well
Plate Microautosampler is immediately aborted but it remains in remote control
and cannot be restarted via a NEXT INJECTION input.
INPUT 1-4: Programmable inputs, can be used in the user program.
E-6
D931R1
User’s Manual FAMOS
Programming Forms
APPENDIX F
The powerful programming features described in Chapter 3 allow the user to set
sampling parameters to meet the precise needs of the analyst. On the following
pages, we provide blank sheets which can be used for hard copy records of
System Menu Settings, Templates, Injection Methods, Wash Methods, Timebase
Methods, Mix Methods and User Programs.
In addition, we provide a form which can be used to describe the FAMOS Well
Plate Microautosampler. This latter form should be faxed to LC Packings when
you are enquiring about your system as it includes information that may assist in
identifying your unit.
Please feel free to photocopy the forms in this section.
User’s Manual FAMOS
D931R1
F-1
Programming Forms
System Menu settings
<GENERAL>
Loop volume
Needle tubing
volume
Syringe
volume
Syringe speed
Needle height
Skip missing
vials
Air segment
Headspace
pressure
Time display
Key click
Error beep
Alarm buzzer
....
....
....
....
<USAGE>
µL
Protection code
µL
Timebase
methods
enabled
disabled
µL
Mix methods
enabled
disabled
User program
enabled
disabled
factor ....
mm
....
no
Labeled wells
enabled
disabled
yes
no
enabled
disabled
yes
no
Templates
Calibration
wells
enabled
disabled
HH:MM:SS
H:MM:mm
on
off
on
off
on
off
processing in:
ROWS
COLUMNS
position first
transport vial:
position last
transport vial:
Off
......
yes
<PLATES>
96-low
96-high
384-low
48-vials
Type:
User program
protection code
......
....
....
<CLOCK>
On (yy/mm/dd and hh/mm)
<IO>
Inject-marker
pulse length
Well-marker
pulse length
Labeled wellmarker pulse
length
Input edge next
injection
Input edge next
well
Freeze input
active
Reset outputs
after last series
....
sec
....
sec
....
sec
falling
rising
falling
rising
low
high
yes
no
<COMM.>
Device identifier:
2….
Comments:
F-2
D931R1
User’s Manual FAMOS
Programming Forms
Templates
Template Injection
Mix
Wash
number method method method
1
Timebase
method
User program
Y/N
Comments
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
Comments:
User’s Manual FAMOS
D931R1
F-3
Programming Forms
Injection Methods
Injection
Method
Number
Type
a full
b partial
c pick-up
Anal.
Time
Flush
Vol.
Inj. /
Well
Injection volumes:
1
2
3
4
5
6
7
8
9
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
Comments:
F-4
D931R1
User’s Manual FAMOS
Programming Forms
Wash Methods
Wash method
number
Wash between
1
injections
wells
series
2
injections
wells
series
3
injections
wells
series
4
injections
wells
series
5
injections
wells
series
Wash volume
Comments:
Comments:
User’s Manual FAMOS
D931R1
F-5
Programming Forms
Timebase Methods
Method number:
Action
AUX 1 1 AUX-1 ON
At Time
Action
ISS-A
1 ISS-A POSITION 6-1
1 AUX-1 OFF
1 ISS-A POSITION 1-2
2 AUX-1 ON
2 ISS-A POSITION 6-1
2 AUX-1 OFF
2 ISS-A POSITION 1-2
3 AUX-1 ON
3 ISS-A POSITION 6-1
3 AUX-1 OFF
3 ISS-A POSITION 1-2
4 AUX-1 ON
4 ISS-A POSITION 6-1
4 AUX-1 OFF
4 ISS-A POSITION 1-2
AUX 2 1 AUX-2 ON
SSV
1 SSV PORT:
1 AUX-2 OFF
2 SSV PORT:
2 AUX-2 ON
3 SSV PORT:
2 AUX-2 OFF
4 SSV PORT:
3 AUX-2 ON
5 SSV PORT:
3 AUX-2 OFF
6 SSV PORT:
4 AUX-2 ON
7 SSV PORT:
4 AUX-2 OFF
8 SSV PORT:
AUX 3 1 AUX-3 ON
CODE
At Time
1 CODE-OUT:
1 AUX-3 OFF
2 CODE-OUT:
2 AUX-3 ON
3 CODE-OUT:
2 AUX-3 OFF
4 CODE-OUT:
3 AUX 3 ON
5 CODE-OUT:
3 AUX 3 OFF
6 CODE-OUT:
4 AUX 3 ON
7 CODE-OUT:
4 AUX 3 OFF
8 CODE-OUT:
AUX 4 1 AUX 4 ON
1 AUX 4 OFF
2 AUX 4 ON
3 AUX 4 OFF
3 AUX 4 ON
3 AUX 4 OFF
4 AUX 4 ON
4 AUX-4 OFF
END
END OF TIMED EVENTS
Comments:
F-6
D931R1
User’s Manual FAMOS
Programming Forms
Mix Method
Method number:
Line
Action
Value
Position
Speed
Height
Line
1
41
2
42
3
43
4
44
5
45
6
46
7
47
8
48
9
49
10
50
11
51
12
52
13
53
14
54
15
55
16
56
17
57
18
58
19
59
20
60
21
61
22
62
23
63
24
64
25
65
26
66
27
67
28
68
29
69
30
70
31
71
32
72
33
73
34
74
35
75
36
76
37
77
38
78
39
79
40
80
Action
Value
Position
Speed
Height
Comments:
User’s Manual FAMOS
D931R1
F-7
Programming Forms
User Program
Line
Action
Value
Position
Speed
Height
Line
1
41
2
42
3
43
4
44
5
45
6
46
7
47
8
48
9
49
10
50
11
51
12
52
13
53
14
54
15
55
16
56
17
57
18
58
19
59
20
60
21
61
22
62
23
63
24
64
25
65
26
66
27
67
28
68
29
69
30
70
31
71
32
72
33
73
34
74
35
75
36
76
37
77
38
78
39
79
40
80
Action
Value
Position
Speed
Height
Comments:
F-8
D931R1
User’s Manual FAMOS
Programming Forms
User Information
Name of user
Company
Department
Address
Telephone
Telefax
FAMOS Well Plate Microautosampler Information
Serial number
Firmware version
Purchase date
Installed options
Local dealer
Address
Telephone
Fax
Service engineer
Address
Telephone
Fax
Comments:
User’s Manual FAMOS
D931R1
F-9
Programming Forms
[This page intentionally left blank]
F-10
D931R1
User’s Manual FAMOS
Connecting the NANO Column
(or other Fused Silica Capillary)
APPENDIX G
G.1
Introduction
These directions should be used when a NANO column or a fused silica capillary
is to be connected to the inert (PAEK) injection valve of the FAMOS Microautosampler or the inert (PAEK) switching valves of the Switchos II Advanced
Microcolumn Switching Unit.
CAUTION
Caution: Do not use a stainless steel nut and/or ferrule with inert (PAEK)
injection/switching valves. The use of stainless steel nuts or ferrules may
damage the valve. Use only the supplied fittings (PEEK) and follow the
instructions below.
G.2
Using PEEK Fingertights
The PEEK sleeve connection is created using the components shown in
FIGURE G-1:
• PEEK fingertight nut,
• PEEK ferrule,
• PEEK sleeve (with an appropriate I.D.),
• NANO column (or any other fused silica capillary)
• The female fitting port of the inert valve
(FIGURE G-1 shows a stainless steel union, which is used only for the?
assembly of the connection).
Union
Sleeve
Fingertight Nut
NANO Columnn
Ferrule
FIGURE G-1 Parts of a PEEK sleeve connection
User’s Manual FAMOS
D931R1
G-1
Connecting the Nano Column
G.3
Fitting Assembly
To assemble the PEEK sleeve fitting (using the supplied stainless steel union):
a) Slide the fingertight nut and the ferrule onto the sleeve as shown in
FIGURE G-1.
b) Insert this assembly into the union that is provided, screwing the nut in two
or three turns by hand.
c) Push the sleeve all the way forward into the union so that the sleeve seats
firmly. This is essential for a proper zero dead volume connection!
FIGURE G-2 Attaching a PEEK Sleeve connector to a NANO column
d) Manually turn the nut into the union until it is finger tight. Carefully turn the
nut 1/4 turn (90°) past the point where the ferrule starts to grab the sleeve
(FIGURE G-2). Use a ¼” wrench to retain the union.
e) Remove the pre-assembled sleeve fitting and inspect it. The ferrule should be
firmly attached to the tubing (i.e. you should not be able to move it laterally
along the tubing axis). If the ferrule is loose, reinstall the tubing in the fitting
in the union and tighten it another 1/8 of a turn past finger tight.
f)
Push the NANO column (capillary) all the way forward into the pre-assembled
sleeve so that the sleeve and column seat firmly. This is essential for a
proper zero dead volume connection!
Note: It is essential that the sleeve and the NANO column (capillary) are inserted
all the way forward into the union for a proper zero dead volume connection!
g) Manually turn the nut into the union until it is finger tight. Carefully turn the
nut 1/4 turn (90°) past the point where the pre-assembled sleeve first start
to grab the tubing (FIGURE G-2). Use a ¼” wrench to retain the union.
Note: The amount of force to turn the nut can vary considerably due to the
friction between the nut and the threads and as well as the composition and wall
thickness of the tubing used. Because of these variables, we do not provide a
torque specification.
G-2
D931R1
User’s Manual FAMOS
Connecting the Nano Column
h) Remove the fitting and inspect it. The PEEK sleeve should be firmly attached
to NANO column or the capillary, respectively (i.e. you should not be able to
move it laterally along the tubing axis). If the PEEK sleeve is loose, reinstall
the fitting in the union and tighten it another 1/8 of a turn past finger tight.
i) Remove, re-inspect, and repeat, if necessary.
FIGURE G-3 Installing the Pre-assembled Fitting
j)
CAUTION
Install the pre-assembled fitting in the inert (PAEK) injection/switching valve
and manually turn the nut into the valve until it is finger tight (FIGURE G-3).
Caution: Never use the inert (PAEK) injection/switching valves to pre-assemble a
fitting and never use any tool to tighten the fingertight nut. This may lead to a
damage of the inert (PAEK) valve.
G.4
Using Long PEEK Hex Style Nuts
In case the supplied fingertight fittings cannot be used due to limited space, use
the long PEEK hex style nuts (P/N 161007) supplied with the instruments instead
(FIGURE G-4).
FIGURE G-4 The Long PEEK Hex Style Nut
To use the supplied long hex nuts:
a) Assemble the fitting as described in Section G.3.
b) Install the pre-assembled fitting in the inert (PAEK) injection/switching valve
and manually turn the nut into the inert (PAEK) valve until it is finger tight
(FIGURE G-3).
c) Carefully tighten the hex style nut using a ¼” wrench.
CAUTION
Caution: Never use the inert (PAEK) injection/switching valves to pre-assemble a
fitting and never use any tool to tighten the fingertight nut. This may lead to a
damage of the inert (PAEK) valve.
User’s Manual FAMOS
D931R1
G-3
Connecting the Nano Column
G.5
Spare Parts Lists
G.5.1
Tubing and Fittings
Part Number
161000
161007
G-4
Description
PEEK Fingertight nuts and ferrules for inert injection/switching valves,
set of 10
PEEK Hex nuts and ferrules for inert injection/switching valves, set of
10
D931R1
User’s Manual FAMOS
Index
Note: Items indicated in Brackets and Capital Letters Refer to Commands on the
Display
<µL PICK-UP> 3-18
µL pick-up Injections 1-2
Air Segmentation A-8
Standard Mode A-6
Connections (Electrical) 2-5
Auxiliaries (P5) 2-6, E-3
Communication 2-6
Markers (P4) 2-6, E-2
Outputs E-2
Power 2-7
Contact Closure E-2
Contents (of Manual) 1-4
<COOL> 3-12
Cooling 3-12
Testing 5-6
<CONVENTIONAL> 3-13
<COPY> 3-11
<COUNTS> 3-11
Cursor Keys
(see: Arrow Keys 3-3)
A
<AIR SEGMENT> 3-14
Air Segmentation
µL Pick-Up Injections A-6
Full Loop Injections A-3
Partial Loop Injections A-5
<ALARM BUZZER> 3-14
<ANALYSIS TIME> 3-18
Arrow Keys 3-3
<ASPIRATE>
Mix Method 3-21
User Program 3-22
<AUTOMATIC> 3-12
<AUX>
Timebase 3-20
User Program 3-24
Auxiliaries (P5 Connector) 2-6
D
<DATE-TIME> 3-12
<DEFAULT ALL> 3-11
<DELETE>
Mix Method 3-21
User Program 3-22
<DESTINATION> C-7
Device Identifier (COMM Menu) 3-17
Digital Inputs and Outputs E-1
<DISPENSE>
Mix Method 3-21
User Program 3-23
Display 3-3
Drain Wash-position 2-9
B
Buffer Tubing 2-3, 2-4, 6-7
C
<CALIBRATION WELLS> 3-15
CE-mark 2-5
CE Declaration xi
Checking
Loop Volume 5-5
Syringe Accuracy 5-5
Tray Cooling (Option) 5-6
CL (Clear key) 3-3
<CLOCK> 3-17
<CODE>
Timebase 3-20
User Program 3-24
Column Switching C-12
<COMM.> 3-17
Communication Connector 2-6
<COMPRES>3-24
Condensed water and leakage 2-9
Configuration
Standard 2-3
Injection < 1 µL 6-7
Injection > 200 µL 6-8
Standard Injection 6-8
Connecting to an HPLC System 2-8
User’s Manual FAMOS
E
E (Enter key) 3-3
<EDIT>
Mix Method 3-21
User Program 3-22
END TIME <3-20>
<ERASE> 3-11
Electrical Connections 2-5
Entering a Program 3-6
<ERROR BEEP> 3-14
Error Codes 6-11
Escape (Key) 3-4
<EXCHANGE> 3-10
<EXIT> 3-12
Executing a Program 3-6
Executing a Series 3-8
Remote Control 3-9
D931R1
I-1
Index
F
Factory Installed Items 2-3
Fan 2-5
Features of System 1-1
<FIRST TRANSPORT VIAL> 3-16
<FLOW RATE> 3-19
Fluid Connections 2-8, 2-9
<FLUSH VOLUME> 3-18
<FREEZE INPUT ACTIVE> 3-16
Front Panel 3-3
<FULL LOOP> 3-18
Full Loop Injections 1-2, A-1
Function Keys 3-3
Fused silica needle 6-4
Fuses (replacing) 6-8
<KEY CLICK> 3-14
Keypad 3-3
Keys 3-3
L
<LABELED WELLS> 3-15
<LABELED WELL MARKER PULSE
LENGTH> 3-16
<LAST TRANSPORT VIAL> 3-16
<L.D. FACTOR> 3-19
Location in Laboratory 2-1
<LOAD> 3-23
<LOG> 3-11
<LOOP VOLUME> 3-13
Loop Volume 6-7
Testing 5-6
Low Dispersion Injection A-9
G
<GENERAL> 3-13
General Design 1-3
General Menu 3-13
General Settings 3-14
M
Mains Input 2-7
Mains Switch 2-7
Maintenance 6-2
<MANUAL> 3-12
<MARKERS>
User Program 3-24
Markers (P4 Connector) 2-6, E-2
[MENU] 3-5
Menu (Key) 3-4
Methods (Key) 3-4
Methods (Menu) 3-5, 3-17
<METHODS>
<INJECTION> 3-18
<MIX> 3-21
<TEMPLATE>3-25
<TIMEBASE> 3-20
<USER PROGRAM> 3-22
<WASH> 3-19
Methods Menu 3-5, 3-17
Method Types 3-7
<MICRO> 3-13
<MIX> 3-21
Mix Menu 3-21
Mix Method 3-7
enable/disable 3-5
<MIX METHODS> 3-15
Mobile Phase Considerations 2-12
[MN] 3-5
H
<HEADSPACE PRESSURE>3-14
Help (key) 3-4
Hold/Cont. (key) 3-3
<HOME> 3-23
HPLC connections 2-8
I
Initialization 3-2
<INJECTION> 3-18
Injection Principle 1-2, A-1
Injection Method 3-7, 3-18
number 3-5
<INJECTION CONTROL> 3-19
<INJECTION VOLUME> 3-19
<INJECT-MARKER PULSE LENGTH> 3-16
INPUTS 2-5
TTL E5
<INPUT EDGE NEXT INJECTION> 3-16
<INSERT>
Mix Menu 3-21
User Program 3-22
Installation 2-1
Plate 4-3
Interrupt (key) 3-4
Introduction
(see: System Overview) 1-5
IO Menu 3-16
IO connections 2-5
IO Defaults 3-17
ISS
Timebase 3-20
User Program 3-23
N
Needle
Height, Adjusting 2-11
Optional 6-5
Prepuncturing 6-4,6-6
Replacing 6-4
Sample 6-4
<NEEDLE> 3-23
<NEEDLE HEIGHT 3-14
K
I-2
D931R1
User’s Manual FAMOS
Index
<NEEDLE TUBING> 3-13
<NO> 3-25
<NUMBER OF INJECTIONS PER
VIAL> 3-19
Numeric Keys 3-3
Safety Precautions ix
Sample
Considerations 2-12
Handling 2-12
Injector D-1
Injection Modes
Comparison A-10
Sample loop 6-7
Sample Needle
Replacing 6-4
<SCALE FACTOR> 3-13
<SERIAL> 3-12
Series (Key) 3-4
Series Menu 3-5,3-26
without templates 3-26
with templates 3-26
<SERVICE> 3-12
Silica-coated Needle 6-4
<SKIP MISSING VIALS> 3-14
Soft Function Keys 3-3
Spare Parts 6-13
Specifications 7-1
<SSV>
Ready Menu 3-12
Timebase Program 3-24
Standard System Configuration 2-3, 6-7
Start/Stop (key) 3-3
<SYR> 3-23
<SYR END> 3-10
<SYR HOME> 3-10
Syringe
replacing 6-3
types of 6-4
waste 2-9
Accuracy (Checking) 5-5
Syringe Speed 3-25
<SYR_VALVE> 3-23
<SYRINGE SPEED> 3-13
Syringe Tip
Replacing 6-4
<SYRINGE VOLUME> 3-13
Syringe Waste Tubing 2-9
System (Key) 3-4
System Menu 3-5, 3-13
System Settings 4-2
O
Outputs 2-5
Contact Closure E-2
Timed E-4
TTL E-4
P
<PANIC> 3-12
<PARTIAL LOOPFILL> 1-2, 3-18
Partial Loopfill A-4
Standard Mode A-4
Air Segmentation A-5
Peltier Cooling 1-1, 3-12
Plates
Handling 2-12
Menu 3-16
<PLATES> 2-13, 3-10, 3-16
Plate Holder
Installation 2-8
<PLATE HOME> 2-13, 3-10
Power Connector 2-7
Powering Up 3-2
Preparations for Use 2-5
Prepuncturing Needle 6-4
Replacing 6-6
Principle of Operation 1-2
Priority (key) 3-4
Priority Sample 3-4
<PROG-OUT> 3-24
Programming
Charts B-1
Examples 4-1, C-1
Forms F1
<PROTECTION CODE> 3-15
R
Ready Menu 3-2, 3-5, 3-10
Reagent Vials (placing in position) 2-13
Rear Panel 2-5
<REMOTE> 3-9
Remote Control 3-9
<REPEAT>
Mix Method 3-22
Replacing
Needle Assembly 6-4
Prepuncturing Needle 6-6
Syringe 6-3
Syringe Tip 6-4
Returning Instruments vii
T
Temperature Control 3-12
<TEMPLATE> 3-15
Template
Definition 3-7
Create 3-25
Enable/disable 3-15
Menu 3-25
Testing 5-1
Test Protocol 5-2, 5-3
<TIMEBASE> 3-20
S
User’s Manual FAMOS
D931R1
I-3
Index
Timebase method 3-7, 3-20
<TIME DISPLAY> 3-14
Timed Outputs E-4
Transport
liquid 1-2, A-3
Vials (first, last) 3-16, 4-7
TTL Inputs E-5
TTL Outputs E-4
Tray Cooling
Testing 5-6
Troubleshooting 6-10
<TYPE OF PLATE>3-16
U
<UNLOAD> 3-23
Unpacking 2-2
Usage Menu 3-15
<USER PROGRAM> 3-15, 3-22
User Interface 3-1
User Program 3-7
enable/disable 3-15
<UTILS> 3-11
V
Valco Valve D-1
<VALVES>
Timebase Method 3-20
User Program 3-23
Voltage Selector 6-8
W
<WAIT>
Mix Method 3-22
User Program 3-24
<WAIT-IN> 3-24
Warnings ix
Warranty v
<WASH> 2-11
Mix Method 3-22
Ready Menu 3-10
User Program 3-23
Wash Method 3-19
Wash Menu 3-19
Wash Method 3-7
Wash Solvent Bottle 2-10
<WASTE> 3-23
Waste Tubing 2-9
<WELL-MARKER PULSE LENGTH> 3-16
<WELL PROCESSING METHOD> 3-16
Wells see Plates
Y
<YES> 3-25
I-4
D931R1
User’s Manual FAMOS