0060.170-11 Optimas Manual PDF

May 2014
OPTIMAS™ manual
Document no. 0060.170-11
BETTER SAMPLE CARE

Other products by Spark Holland B.V.:
 Alias™ Autosampler
 Integrity™ Autosampler
 Mistral™ Column Thermostat
 Reliance™ Autosampler
 Symbiosis™ system
 Symbiosis™ Pro system
 Symbiosis™ Pico system
 SPH1240™ Gradient pump
 Hysphere™ SPE cartridges
Spark Holland B.V.

P.O. Box 388
7800 AJ EMMEN
The Netherlands
P. de Keyserstraat 8
7825 VE EMMEN
The Netherlands
P +31 591 631 700

F +31 591 630 035
www.sparkholland.com
OPTIMAS™ is a registered trademark of Spark Holland B.V. All other brands, company and product names are or may be trademarks of their
respective holders and are used to identify their products or services.
Copyright © 2014 Spark Holland B.V. All rights reserved. No part of this publication may be reproduced, stored in a retrieval system or
transmitted in any form by any means, without written permission from Spark Holland B.V.
Spark Holland B.V. reserves the right to make changes to the design and specifications of the system and this manual without prior notice.
0060.170-11 Optimas manual
Contents
1 About this manual 5

1.1 Pictorials 5
1.2 Safety instructions 6
1.3 Typographical conventions 8
2 Introduction 9
3 Identification of parts 11
3.1 Keyboard and display 14
4 Injection principles 17
4.1 Full loop injections 17
4.2 Partial loopfill injections 19
4.3 µL pickup injections 22
5 Using the OPTIMAS 25

5.1 Example - 10 uL Partial loopfill injection from vial number 1 26
5.2 Example - 3 x 10 µL pickup injections 27
5.3 Example - a 1:10 dilution + 10 µL partial loopfill injection 29
6 Programming reference 31
6.1 Programming overview 31
6.2 Ready screen options 33
6.2.1 <WASH> 33
6.2.2 <MAINTENANCE> 34
6.2.3 <TEMP> 34
6.2.4 <UTILS> 35
6.2.5 <SERVICE> 35
6.3 System settings programming 36
6.3.1 <GENERAL> 36
6.3.2 <METHOD> 38
6.3.3 <TRAY> 38
6.3.4 <IO> 39
6.3.5 <PREP> 39
6.4 Method programming mode 40
6.4.1 <INJECTION> 41
6.4.2 <WASH> 42
6.4.3 <MIX> 43
6.4.4 <TIMED EVENTS> 45
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Optimas manual 0060.170-11
6.5 Series programming mode 47

6.6 Run options 48
6.6.1 [Start/Stop] 48
6.6.2 [Priority] 50
6.6.3 [Hold Cont.] 50
6.6.4 Remote control 51
6.6.5 Programming during the run 51
7 Maintenance 53
7.1 Injection valve and rotor seal 53
7.2 Needle and tubing 54
7.3 Sample needle 57
7.3.1 Sample needle replacement 58
7.3.2 Air needle replacement 58
7.3.3 Sample needle height setting 58
7.4 Syringe 59
8 Specifications 61
9 Trouble shooting 65
9.1 Start up problems 66
9.2 Analytical problems 67
9.2.1 No injection 68
9.2.2 Bad reproducibility 69
10 Installation 71
10.1 Fluid connections 74
10.1.1 HPLC connections 75
10.1.2 Waste tubing 75
10.1.3 Wash solvent and system rinse 76
10.1.4 Syringe 77
10.2 Preparation of vials 77
10.2.1 Types of vials 77
10.2.2 Filling and sealing the vials 81
10.2.3 Loading the sample tray 81
10.3 Control I/O connections 81
10.3.1 Contact closures output (P1) 81
10.3.2 TTL input (P2) 82
Appendix A. Spares and accessories 85
Appendix B. Error codes 87
2
Appendix C. Test procedure 89
Appendix D. Prep option 93

Specifications 93
Installation 94
Injections 94
Appendix E. Log book 97

System settings sample 98
Method log sample 99
Appendix F. Storage, packaging, recycling and disposal 101
Index 103
3
4
1 About this manual
This manual is intended for laboratory technicians who use the OPTIMAS
autosampler for execution of analytical runs.
Please read this manual and the safety instructions carefully before you start to use
the autosampler.
This manual provides the following information on the autosampler:
 section 1 offers a short introduction to the autosampler
 section 2 offers identification of parts and a description of the injection
principles of the autosampler
 section 3 offers procedures for use of the autosampler
 section 4 offers maintenance procedures
 section 5 lists specifications
 section 6 offers trouble shooting information
 section 7 describes the installation procedure for the autosampler
 appendix A lists available spares and accessories
 appendix B lists error codes and how to resolve them
 appendix C offers a test procedures
 appendix D describes the Prep option
 appendix E contains sample log book pages for the OPTIMAS
 appendix F describes procedures for storage, packaging and end-of-life.
An index is provided for easy reference.
1.1 Pictorials
The following pictorials are used in this manual:
The danger sign warns about a hazard. It calls

attention to a procedure or practice which, if not
adhered to, could results in injury or loss of life.
Do not proceed beyond a danger sign until the
indicated conditions are fully understood and met.
5
The warning sign denotes a hazard. It calls attention

to a procedure or practice which, if not adhered to,
could result in severe injury or damage or destruction
of parts or all of the equipment. Do not proceed
beyond a warning sign until the indicated conditions
are fully understood and met.
The caution sign denotes a hazard. It calls attention to

a procedure or practice which, if not adhered to, could
result in damage or destruction of parts or all of the
equipment. Do not proceed beyond a cautions sign
until the indicated conditions are fully understood and
met.
The attention sign signals relevant information. Read

this information, as it might be helpful.
The note sign signals additional information. It

provides advice or a suggestion that may support you
in using the equipment.
This symbol indicates that the waste of electrical and

electronic equipment must not be disposed as
unsorted municipal waste and must be collected
separately. Please contact an authorized
representative of the manufacturer for information
concerning the decommissioning of equipment.
1.2 Safety instructions
Adhere to standard quality control procedures and the following equipment

guidelines when using the system:
Working environment & safety
The intended use of the system is to handle HPLC solvents.
Use this system ONLY for the intended use. Use of the
system for any other purpose may cause unsafe situations.
The system is suitable for use in a GLP-approved
environment. If that is the case, operators using the system
should have an extensive understanding of GLP rules.
6
System Operation
To keep up the high performance of the system we
recommend that the system is checked regularly and
maintenance procedures are carried out.
Do not use the device if there is visible damage.
Note that the OPTIMAS is a high-pressure device. Always
make sure pressure in the device is built up slowly. Reduce
the pressure gradually to 0 before disconnecting any tubing
or before opening the purge valve. Sudden loss of pressure
may damage the pump and the analytical column connected
to it.
Electrical safety
Removal of panels may expose users to dangerous voltages.
For that reason this should only be done by authorized
service engineers. Disconnect the system from all power
sources before removing protective panels.
Replace or repair faulty insulation on power cords
immediately after discovery of the fault.
Check that the actual power voltage is the same as the
voltage for which the instruments are wired. Make sure power
cords are connected to correct voltage sources.
The system must only be used with appliances and power
sources with proper protective grounding to prevent damage
through buildup of static electricity.
Do not place the system in the vicinity of equipment that emit
EMC radiation.
Solvents
The solvents used may be flammable and toxic. The room in
which the system is installed should be well ventilated to
prevent that solvent vapors cause poisoning or ignite and
cause a fire.
Use of open fire in the vicinity of the system must be strictly
prohibited. Do not install the system in the same room with
any other equipment that emits or could potentially emit
sparks.
Provide protective equipment near the instrument. If solvent
gets into the eyes or on the skin, it must be flushed away
immediately. Provide equipment, such as eye wash stations
and safety showers, as close to the system as possible.
Solvents bottles should be sealed to minimize any risks
related to solvent vapors.
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.
Perform periodic leak checks on supply lines.
7
Biohazard
When you analyze biological fluids you need to take all
possible precautions and treat all specimens as potentially
infectious. Always wear protective goggles and gloves when
handling toxic or biologically infectious samples to prevent
bio hazards or toxic hazards while working with the
OPTIMAS.
1.3 Typographical conventions
The following typographical conventions are used to refer to the various controls
and display text of the OPTIMAS autosampler:
control keys and Start/Stop, in square brackets.
Hold/Cont., keys of numerical Examples: [Start/Stop], [Escape]
keypad
function keys in pointed brackets, just like in the

display. Examples: <WASH>, <SYR
END>
8
2 Introduction
OPTIMAS has been designed to meet the challenge of the modern analytical
laboratory: it is robust, cost-effective and easy to implement. Sample cooling is
available for consistent results. High-resolution syringe control guarantees superior
precision for injection and reagent addition. The use of these and many more
features is described in this manual to help you exploit the full potential of
OPTIMAS.
PASA™, a robust injection concept

Loop injection with Pressure Assisted Sample Aspiration (PASA™) is a proven
concept that combines high precision with simplicity and reliability. No moving
around with the sample needle. Reduced risk for bubbles in the sample line. No
needle port that wears and contaminates. There is only intelligent valve switching
and highly accurate syringe control. Yet, three injection modes can be selected: full
loop, partial loop filling and µL Pick-up for maximum precision, maximum flexibility
and zero sample loss.
Figure 1: The PASA™ injection concept
Side-port needle: a good point!

The strong side-port needle combines the optimum point style for piercing with a
minimum risk of blockage by septum particles. For small sample volumes, a spring-
loaded needle is available for maximum sample aspiration independent of
variations in vial dimensions.
9
Integrated sample cooling option

Thermo-electric sample cooling is available as a fully integrated option. No external
device is required and the efficient cooling concept does not restrict access to the
sample tray during operation.
Reagent addition made simple

Internal Standard addition, sample dilution or derivatization can be programmed in
a very simple manner. A single-stage derivatization of a sample in a separate
(destination) vial requires no more than 4 program lines. Multi-reagent addition is
also possible: two large volume reagent vials are available on the sample tray.
Service friendly design

Low instrument down time is accomplished by a high Mean Time Between Failure
and quick instrument service.
10
3 Identification of parts
OPTIMAS is a standalone autosampler of robust design. This section describes

controls and parts that are important for correct use of the autosampler.
Proceed as follows for access to the sampling compartment:
1. Hold the cover with both hands on either side of the autosampler.
2. Carefully lift the cover (see figure 2).
3. Rest the cover on the top of the autosampler.
Figure 2: Access to sampling compartment
11
The following parts are important for use of the autosampler:
Figure 3: Main parts of the autosampler
1. Needle arm
2. Injection valve
3. Wash position
4. Position for transport solvent and reagent vials
5. Wash and waste drain
6. Condensed water and leakage drain
7. Tray fastening knob
8. Sample tray
9. Syringe dispense
10. Display and keyboard (on cover)
12
The back of the autosampler offers the following:
Figure 4: Back of the autosampler
1. CE label
2. I/O connector (inputs)
3. Fan (only when tray cooling option is installed)
4. Fuse box and voltage selector
5. Power connector
6. On/off switch
7. I/O connector (outputs)
13
3.1 Keyboard and display
The OPTIMAS is controlled by way of a keyboard and a display:
Figure 5: Display and keyboard
1. Display
2. Function keys (see below for more information)
3. Numerical keypad (see below for explanation)
4. Start/Stop button (see below for explanation)
5. Hold/Continue button (see below for explanation)
6. Control keys (see below for explanation)
Explanation of keyboard keys:

 Function keys: these keys refer to the function that is displayed directly above
the key concerned. The display will show the key name in <...>; the function of
the key depends on the status of the OPTIMAS. Please note that only four
functions will be displayed; if more functions are available, [MENU] will appear
in the top right corner of the display. Figure 3-4 above displays three function
keys: for <WASH>, <MAINTENANCE> and <TEMP>. As the display also
shows [MENU], you can press the [Menu] control key to show other available
function keys.
 Six general control keys to:
[PRIORITY] Interrupt a sample run to process an emergency
sample prior to the next sample of the
programmed sample Series.
After the priority sample the sample run is
continued.
[SERIES] Enter the Series programming mode. In the
Series, vials are linked to methods. See section
6.5 for more information.
[METHODS] Enter the Methods programming mode. See
section 6.4 for more information.
14
[SYSTEM] Enter the System Settings programming mode. In

the system settings the configuration of the
autosampler is defined. See section 6.3 for more
information.
[ESCAPE] Return to the previous programming level when
you are in programming mode, the programmed
parameters are checked and stored in battery
backup memory.
[MENU] Display other available function keys. This key is
only active when "[MENU]" appears in the right
top corner of the display (see figure 5); it allows
you to navigate to other available function keys.
 Numerical keypad:
[CL] Clear; Removes the actual value of the active
programmable parameter. In some cases the
value NONE will be displayed.
[E] Enter; Stores and checks the values in the
display. If all values are valid the next
programming screen will be displayed.
[0-9] Numeric keys; used to enter values for
programming parameters. Press [E] to store
values; press [ESCAPE] to leave the
programming mode.
 [START STOP]: Start and stop automatic processing.
 [HOLD CONT.]: Hold or continue the countdown of the analysis time. The
analysis time is extended with the time that Hold is active.
15
16
4 Injection principles
A unique sequence of valve switching and syringe dispenser controlled sample

withdrawal enables the OPTIMAS to inject sample volumes in a range from a few
microliters to milliliters in a simple and reproducible manner.
Three different methods for injection can be selected:
Full loop: The sample loop is completely (quantitatively) filled with

sample, resulting in extremely good reproducibility.
Partial loopfill: The sample loop is partially filled with sample; this means
low sample loss and programmable injection volumes.
µL pick-up: After aspiration from the vial the sample volume is
transported into the loop with transport liquid (mobile
phase) from another vial; no sample loss.
The OPTIMAS uses a syringe to aspirate the sample from a vial into the sample
loop. To prevent contamination of the syringe the OPTIMAS is equipped with a
buffer tubing between the syringe and the injection valve.
The wash solvent is used to remove the sample from the buffer tubing and sample
needle and rinse the buffer tubing and sample needle.
4.1 Full loop injections
The switching sequence for a full loop injection is schematically shown below:
1. The Initial situation: the injection valve is in the INJECT position. The sample
needle with air needle has entered the vial. The headspace pressure, applied
through the outer air needle, ensures that no air or vapor bubbles are formed
during sample aspiration.
17
2. The syringe dispenser aspirates the "flush volume" from the sample vial to fill
the sample line with sample and remove wash solvent.
3. The injection valve switches to the LOAD position, placing a distinct sample
plug at the inlet of the sample loop.
4. The sample loop is quantitatively filled by transporting a number of times the

loop volume through the loop, depending on the volume of the loop:
3x loop volumes for loops <= 100 µL
2x loop volumes for loops from 100 µL to 500 µL
1½x loop volumes for loops > 500 µL
5. The injection valve switches into the INJECT position. The sample loop is now
part of the HPLC mobile phase flow path: sample is transported to the column.
The analysis starts.
18
If one injection is to be made out of each vial or a wash routine has to be

performed after every injection: the needle withdraws from the vial immediately
after the injection and, if programmed, immediately performs a wash. After the
analysis time has passed, a new sequence is started.
The next injection sequence will start with a flush of 50% of the programmed flush
volume, in case an injection from the same vial and no wash routine is
programmed. Otherwise it will start with a flush of the programmed flush volume. If
the withdrawal of sample for the next injection will exceed the total volume of the
sample buffer tubing, the buffer tubing is rinsed before the next injection. The next
injection will start with the programmed flush.
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 and will not be injected and therefore will
not influence the injection. The air segment may be selected in the system settings
(ON/OFF, see section 6.3.1).
With a standard needle, the flush volume must be at least 30 µL for injections with
air segment and 35 µL for injections without air segment. If the samples are highly
viscous it may be necessary to program larger flush volumes and reduce the
syringe speed for better performance.
Figure 6: Situation after filling the loop (A=with air segment; B=with air segment)
4.2 Partial loopfill injections
The switching sequence for a partial loopfill injection is schematically shown in the
following figures:
19
needle with air needle has entered the vial. The headspace pressure, applied
through the outer air needle, ensures that no air or vapor bubbles are formed
during sample aspiration.
2. The syringe dispenser aspirates the "flush volume" from the sample vial to fill
the sample line with sample and remove wash solvent.
3. The injection valve switches to the LOAD position, placing a distinct sample
plug at the inlet of the sample loop.
4. The programmed injection volume is now aspirated into the sample loop.
20
5. The injection valve switches to the INJECT position. The sample loop is now
The next injection sequence will start with a flush of 50% of the programmed flush
volume, in case an injection from the same vial and no wash routine is
programmed. Otherwise it will start with a flush of the programmed flush volume. If
the withdrawal of sample for the next injection will exceed the total volume of the
sample buffer tubing, the buffer tubing is rinsed before the next injection. The next
injection will start with the programmed flush.
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 and will not be injected. The air segment
can be selected in system settings (ON/OFF, see section 6.3.1).
With a standard needle the flush volumes must be minimal 30 µL for injections with
air segment and 35 µL for injections without air segment. If the samples are highly
viscous it may be necessary to program larger flush volumes and reduce the
syringe speed for better performance.
Figure 7: Situation after filling the loop (A=with air segment; B=without air segment)
21
4.3 µL pickup injections
The switching sequence for a µL pick-up injection is schematically shown in the

following figures:
needle has entered the vial of transport liquid (mobile phase, this to avoid
disturbance of the chromatogram with an additional peak of the transport
solvent).
2. For the first injection, after a wash or after emptying of the buffer tubing. The
syringe dispenser aspirates transport liquids from the transport vial to fill the
sample line with transport liquid and remove wash solvents.
3. The needle moves from the transport vial to the sample vial. The injection
valve switches to the LOAD position.
4. The programmed injection volume is aspirated from the sample vial.
22
5. The sample needle moves back to the transport vial. The sample is
quantitatively transported into the loop, with transport liquid (mobile phase)
from the transport vial.
6. The injection valve switches into the INJECT position. The sample loop is now
For the next sequence the first withdrawal of transport solvent is omitted, unless a
wash routine is performed or the OPTIMAS has emptied the buffer tubing to the
waste position. In these cases the sequence is completely repeated.
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. An air segment can be
programmed in the system settings (see section 6.3.1).
23
In this mode,the air segment at the front of the sample

plug is injected into the HPLC system.
Figure 8: Situation after filling the loop (A=with air segment; B=without air segment)
In case of µL pick-up injections, there will be no air

pressure (headspace pressure) on the vials, to avoid
sample errors due to air expansion during vial exchange
from the sample vial to the transport solvent vial.
24
5 Using the OPTIMAS
Use this section to get started with the OPTIMAS, after the installation instructions
(see section 10) have been executed.
Three different examples are described below; make sure you adapt values for the
type of application you wish to use.
Installation (for more detailed information see section 10 of this manual):
1. Allow the OPTIMAS to adopt ambient temperature for at least one hour
2. Check OPTIMAS for visual damage resulting from transport; if this is the case,
please contact your distributor immediately.
3. Preferably, place the OPTIMAS autosampler to the left-hand side of your

HPLC system.
4. Remove the safety screw on the right-hand side of the front cover.
Always keep the keyboard front cover closed during
operation.
1. Check fuses and voltage settings on the rear side of the instrument.
2. Connect the power cable.
3. After turning "ON" the power, the Ready screen appears displaying the
firmware revision number.
4. Connect the drain tubing to the wash outlet on the lower front side of the
instrument.
5. Fill the wash solvent bottle with distilled water/ 2-Propanol (80/20 v/v%) or
mobile phase. Only water/organic solvents should be used. Do not use any
crystalline or buffer solutions; these may block the system and cause severe
damage. To avoid air bubbles in the syringe, degas the wash solvent.
6. Fill the wash solvent tubing, syringe and buffer tubing by washing the system 2
or 3 times (press the <WASH> function key from the Ready screen). Check if
there are air bubbles trapped in the syringe; they can be removed by gently
tapping the syringe.
7. Connect the HPLC pump to port 1 of the injection valve.
8. Connect the column (or capillary) to port 6 of the injection valve.
9. Check for leakage and let the system equilibrate for at least 5 minutes.
25
System settings (see section 10 of this manual)

The following settings are standard programmed; to check these settings, proceed
as follows:
Press keys: Description:

[System] Enter the System Settings
<GENERAL> Enter the General Settings
[0100] [Enter] Volume of installed loop: 100 µL.
[015] [Enter] Volume of tubing needle <-> valve:015 µL
<250> [Enter] Syringe volume: 250 µL
<NORMAL> [Enter] Set Syringe speed to Normal.
<YES> [Enter] Skip missing vials.
<YES> [Enter] Air segment: use air segment
<YES> [Enter] Headspace pressure: On
Press [Escape] twice to go back to the Ready screen
Performing injection routines is easy with OPTIMAS:
1. First you define a method such as the injection, mix or wash method. These
methods are stored in battery back up.
2. Subsequently a Run sequence or Series can be programmed in which those

methods are linked to the Series of vials.
3. You can then execute those Series with OPTIMAS.
5.1 Example - 10 uL Partial loopfill injection from vial number 1
This example describes how to execute a 10 uL partial loopfill injection.
Programming the method (see also section 6.4 of this manual)

Injection Program:
[Methods] [1] [Enter] Enter Method number 1.
<INJECTION> <PARTIAL> [Enter] Selecting partial loopfill injection
program
[30] [Enter] Flush volume: e.g. 30 µL
[1] [Enter] Number of injections per vial: 1
[10] [Enter] Injection volume: 10 µL
[100] [Enter] Enter the analysis time e.g. 1 minute
Press [Escape] once to go up one level.
Wash Program:
26

<WASH> Selecting wash program
<NONE> Disable the wash
Press [Escape] three times to go back to the Ready screen.
Programming the Series (see also section 6.5 of this manual)

[Series] [1] [Enter] Enter Series number 1.
[1] [Enter] Enter Method number:1
[1] [Enter] Enter First sample vial: 01
[1] [Enter] Enter Last sample vial: 01
Press [Escape] twice to go back to the Ready screen
Running the Series (see section 6.6 of this manual)

Place a sample vial in position 1 (If you cannot reach position 1, turn the tray
manually).
[Start/Stop]
[1] [Enter] Start at Series number:1
[1] [Enter] Stop after Series number: 01
If you press <START> OPTIMAS will start searching for vial 1 and perform a 10 µL
injection.
5.2 Example - 3 x 10 µL pickup injections
This example describes execution of three 10 µL injections from one vial without
sample loss using the µL Pick-up injection routine, followed by a wash routine
between each injection.
If the Air segment is set ON in System Settings, a small air
segment will also be injected into your HPLC system (for
more detailed information, see section 4.3 of this manual).
For switching OFF the air segment enter the System

Settings and change the setting for the air segment: from
YES to NO.
This is done by pressing:
[System] <GENERAL> [Enter] [Enter] [Enter] [Enter]
[Enter] <NO> [Escape] [Escape]
27

Injection program:
<INJECTION> <PICK UP> [Enter] Selecting µL pick-up injection program
Wash Program:
<WASH> Selecting wash program
<INJECTION> [ENTER} Perform a wash after each injection
[1] [Enter] Enter the number of syringe volumes
to be used for washing, e.g. 1 syringe
volume.
Press [Escape] three times to go back to the Ready screen.

Press [Escape] (twice) to go back to the Ready screen
Running the Series (see also section 6.6 of this manual)

Place a sample vial in position 1 and place a vial with transport solvent (mobile
phase) in position 85.
Make sure the transport vial is filled correctly before starting a new Series.
[Start/Stop]
If you press <START> OPTIMAS starts searching for the transport vial and perform
3 injections of 10 µL. After every injection the needle system will be washed with 1
syringe volume of wash solvent.
28
5.3 Example - a 1:10 dilution + 10 µL partial loopfill injection
This example describes how to perform a 1:10 dilution followed by a 10 µL partial

loopfill injection.
The mix method will perform the following: transfer 360 µL from Reagent A to the
destination vial, add 40 µL sample, mix 3 times with 250 µL and subsequently
inject 10 µL.
From the Ready screen:

Injection Program:
<INJECTION> <PARTIAL> [Enter] Selecting partial loopfill injection
program
[30] [Enter] Flush volume: e.g. 30 µL
Wash Program:
<WASH> Selecting Wash program
<NONE> Disable the wash
Mix Program: (see also section 6.4.3 of this manual)

<MIX> Selecting Mix program
<YES> Confirm the use of the Mix program
<INSERT> Insert the first Mix step:
<ADD> [180] [Enter] Add 180 µL Reagent–A to the
Destination vial.
<REAG-A> [Enter]
<DESTINATION> [Enter]
<INSERT> Insert the second Mix step:
<ADD> [180] [Enter] Add another 180 µL Reagent–A to the
Destination vial.
<REAG-A> [Enter]
29

<INSERT> Insert the third Mix step:
<ADD> [40] [Enter] Add 40 µL from the Sample vial to the
Destination vial.
<SAMPLE>[Enter]
<INSERT> Insert the fourth Mix step:
<MIX> [3] [Enter] Mix the contents of the Destination vial
3 times with a volume of 250 µL
[250] [Enter]
Press [Escape] three times to go back to the Ready screen

[2] [Enter] Enter First destination vial: 02
Press [Escape] (twice) to go back to the Ready screen
Running the Series (see also section 6.6 of this manual)

Place a sample vial in position 1, place an empty sealed destination vial in position
2 and place a filled reagent vial in position 86. Make sure that the reagent vial is
filled correctly before starting a new Series (If you cannot reach one of the tray
positions, turn the tray manually).
[Start/Stop]
If you press <START>, OPTIMAS starts searching for vial 86 and transport 180 µL
to the destination vial twice, subsequently 40 µL of sample will be added and after
mixing 3 times a 10 µL injection will be performed.
30
6 Programming reference
This section describes all programming options for the OPTIMAS autosampler.
6.1 Programming overview
This overview gives a short description of all programming options. Please note
that all options are described in more detail in following sections.
Ready screen options:

<WASH> perform a standard wash
<MAINTENANCE> maintenance functions
<TEMP> <TRAY COOL>
<UTILS> <COPY> copy methods
<ERASE> erase methods
<LOG> logbook
<SERVICE> enter service mode
[SYSTEM] options:
<GENERAL> Loop volume
Needle volume
Syringe volume
Syringe speed
Skip missing vials
Air segment
Headspace pressure
<METHOD> Use calibration vials
<TRAY> Tray type
<IO> Inject marker pulse length
Input edge next injection
Freeze input active
Reset output after last Series
<PREP> Select Prep mode; a number of system settings will be
set to a fixed value.
31
[METHODS] options:
<INJECTION> <FULL> Flush volume
No. Injections/vial
Analysis time
<PARTIAL> Flush volume
No. injections/vial
Injection volume
Analysis time
<PICK-UP> No. injections/vial
Injection volume
Analysis time
<WASH> Wash volume
Wash between: injections/vials/series
<TIMED EV.> timed events program:
4 x AUX-1
4 x AUX-2
END TIME
<MIX> Mix program: Add, Mix, Wait
[SERIES] Method number

First sample vial
Last sample vial
Use calibration vials (depends on system settings)
First calibration vial
Last calibration vial
Vials between calibration
First destination vial (if used in mix method)
[START/STOP] <START>
<REMOTE>
[PRIORITY] Sample vial
[HOLD CONT.] Hold the analysis time of the OPTIMAS
32
6.2 Ready screen options
After power up, the OPTIMAS will run an initialization routine in which it performs a
self-test and every item in the autosampler is set to default positions.
The OPTIMAS then displays the Ready screen:
[MENU]
READY (X.XX)
TRAY:--°C
<WASH> <MAINTENANCE> <TEMP>
In the bottom line of the display, control keys are available that can be used. If
[Menu] is displayed in the top right-hand corner, the [Menu] function key can be
used to navigate to other available control keys.
Press [Menu]. The extra function keys appear :
[MENU]
READY (X.XX)
TRAY:--°C
<UTILS> <SERVICE>
At the right-hand side of the screen the tray temperature is displayed (only when
this option is installed).
This section describes all options available from the Ready screen.
6.2.1 <WASH>
The <WASH> option activates the manual wash of the OPTIMAS. All tubing, the
needle and the syringe will be rinsed.
33
6.2.2 <MAINTENANCE>
<MAINTENANCE> offers access to three options for controlling the needle arm,
the syringe and the injection valve:
<NDL HOR> This function is built-in to facilitate the replacement of the

sample needle, air needle and valve. See also section 7.3
for detailed information. Press <NDL HOR> to move the
needle arm horizontally above the inner sample ring.
When the needle arm reaches the front position. It will
return to its home position by pressing <NDL HOME>.
<SYR END> This function facilitates the exchange of syringes and

filling of the wash solvent tubing. Press <SYR END> to
move the syringe to its end position, thereby aspirating
wash solvent into the tubing and syringe. It stops from
where it will return to its home position by pressing <SYR
HOME>, thereby dispensing the wash solvent to waste.
After replacing a syringe, all tubing can be rinsed using
<WASH> (see section 6.2.1).
<VLV LOAD> With this function the injection valve can be switched.
Press <VLV LOAD> to switch the valve from INJECT to
LOAD.
When the valve is in the LOAD position it can be switched
back to INJECT by pressing <VLV INJ>.
It is not possible to leave this function when the valve is in
the LOAD position, first switch back by pressing <VLV
INJ> and then press [Escape].
To leave <MAINTENANCE> and return to the Ready screen press [Escape].
6.2.3 <TEMP>
Press <TEMP> to enter the temperature control screen for sample tray cooling
(optional).
PROGRAMMING TEMPERATURE SETTINGS
<TRAY-COOLING>
34
If the optional tray cooling is installed, it can be programmed after pressing

<TRAY-COOLING>:
PROGRAMMING COOL SETTINGS

COOLING: OFF SETPOINT: 04°C
<ON> <OFF>
Switch the tray cooling on by pressing <ON> and program the new setpoint. After
pressing [Enter] the new tray cooling setpoint will be activated.
Temperature range tray cooling: 4°C - 15°C

Maximum cool capacity: 20°C below ambient
temperature
Press [Escape] to return to the Ready screen.
6.2.4 <UTILS>
<Utilities> offers the following options:
<COPY> allows you to copy a method.
<ERASE> allows you to erase a method.
<LOG> contains a logbook with information on the number of

switching actions of the injection valve and the syringe valve
and on the number of movements of the syringe dispenser.
6.2.5 <SERVICE>
The service mode can only be entered if you have a service entry code. This code
is only known to authorized service engineers.
1. Press [SERVICE].
2. Enter the code with the help of the keys of the numeric keypad.
3. Press [E] on the numeric keypad. The OPTIMAS is now in Service mode.
35
6.3 System settings programming
Press [System] to enter the System Settings. In this menu all system variables of
the OPTIMAS can be set.
SYSTEM SETTINGS [MENU]

SELECT SETTINGS TO DEFINE
<GENERAL> <METHOD> <TRAY> <IO>
Press [Menu] to navigate to additional options:
SYSTEM SETTINGS [MENU]

SELECT SETTINGS TO DEFINE
<PREP>
The System Settings are divided in five sub-sections: to enter a sub-section, press
the corresponding function key. To step through a sub-section press the [Enter]
key.
After all values have been defined correctly within a sub-section, return the menu
screen of the System Settings by pressing [Escape]. Return to the Ready screen
by pressing [Escape] one more time. The values are stored in battery backup
memory.
The following sections describe the programmable system settings, with their
ranges.
6.3.1 <GENERAL>
Press <GENERAL> to enter general system settings.
Setting Range Comment

Volume of installed loop: 0-5000 µL The volume of the installed loop.
The OPTIMAS uses this value for
calculations of injection volumes.
Volume of tubing needle 0-999 µL The volume of the tubing and
valve needle connected to the injection
valve.
The OPTIMAS uses this value for
calculations of flush and transport
volumes.
Syringe volume <250> The volume of the installed
<1000> syringe.
36
Syringe speed: <LOW> The aspirating and dispensing

<NORMAL> speed are programmable in three
<HIGH> steps:
<NORMAL> for samples with a
viscosity almost equal to the
viscosity of water.
<LOW> for samples with a higher
viscosity and <HIGH> for samples
with a lower viscosity compared to
water.
Syringe speed overview:

250 µL syringe 1000 µL syringe
Syringe Aspirate Dispense Aspirate Dispense
speed:
<LOW> 0.31 mL/min 3.4 mL/min 1.25 mL/min 5.5 mL/min
<NORMAL> 0.62 mL/min 3.4 mL/min 2.50 mL/min 5.5 mL/min
<HIGH> 0.94 mL/min 3.4 mL/min 3.75 mL/min 5.5 mL/min
Wash syringe 3.4 mL/min 13.7 mL/min
speed:
Range Comment
Skip missing vials: <YES> Define if the OPTIMAS has to stop if a vial
<NO> is not found or continue with the next
programmed vial.
Program <NO> to stop the OPTIMAS in
case a vial is not found.
Air segment: <YES> Inject a sample with or without an air
<NO> segment between the sample and the
wash solvent. If the air segment is ON
(YES) the OPTIMAS will draw an air
segment of 5 µL before the flush volume
prior to an injection.
The air segment will not be injected in the
analytical system.
In case of µL pick-up the sample will be
preceded by an air segment, which will be
injected in the analytical system (see also
section 4.3).
Headspace pressure: <YES> The OPTIMAS normally uses headspace
<NO> pressure in combination with the syringe
to transport sample in to the loop. If, for
some reason, no headspace pressure can
be used, switch if off by selecting <NO>.
The compressor will always be used in the
wash position of the OPTIMAS. The
accuracy and reproducibility of the
OPTIMAS may decrease if <NO>
headspace pressure is used.
37
6.3.2 <METHOD>
Press <METHOD> to define the use of calibration vials
Range Comment
Use calibration vials: <ENABLED> Determine if calibration vials can
<DISABLED> be used in Series.
6.3.3 <TRAY>
Press <TRAY> to indicate the type of tray to be used:
Range Comment
Tray type: <84+3> Standard tray with 84 vials, 1.5 mL
and 3 vials of 10 mL for special use.
Vials 85-87, will be used for

transport solvent for the µL pick-up
injections.
When the use of mix item is
enabled in the System Setting the
three 10 mL vials will be used as
follows:
vial 85: transport solvent
vial 86: Reagent A
vial 87: Reagent B
<96> Tray with 96 vials of 1.5 mL
<24> Tray with 24 vials of 10 mL
Vial type: <STANDARD> Standard vials are used
(only when <2.5ml> only 2.5 mL vials are used.
<84+3> or <96> is (Chromacol 2.5-CV, 12 X 40 mm)
selected)
Only with the standard <84+3> tray it is possible to

perform a mix and to use the µL pick-up injection mode.
With the other two trays both the mix and the µL pick-up
injection mode will automatically be disabled.
If 2.5 mL vials are used be sure to fill the vials completely

before starting a RUN.
38
6.3.4 <IO>
Press <IO> to configure Inputs and outputs:
Range Comment (see also section

10.1)
Injection marker pulse 0.0 - 2.0 sec. Define the length of the inject
length: marker pulse. Normally 1.0 sec.
Input edge next <FALLING> Define the edge sensitivity of the
injection: <RISING> next injection input.
Freeze input active: <LOW> Define the level to activate the
<HIGH> freeze function.
If the freeze input is set to <HIGH>, the time base will

freeze if the input is not connected to another device.
Consequently, the OPTIMAS will not start!
Reset outputs after last <YES> Define if the outputs should be

Series: <NO> reset to the default settings after
finishing the last Series.
6.3.5 <PREP>
Press <PREP> to enter settings for the Prep Mode (optional; see also Appendix D):
Range Comment
Use Prep Mode: <YES> Define if the OPTIMAS is a standard
<NO> autosampler, or a Prep mode
autosampler.
When choosing for the Prep Mode, some System

Settings are no longer programmable, but set to a fixed
value. See also Appendix D.
In the Prep Mode the mix, the full loop and µL pick-up
injection mode, will automatically be disabled.
39
In the Prep Mode not all of the System Settings are programmable. see table below
for an overview.
Fixed System Settings in Prep Mode:

General System Settings
Volume of the installed loop 10000 µL
Syringe volume 2500 µL
Air segment <NO>
Method System Settings
Use of calibration vials <DISABLED>
Tray System settings
Tray type <24>
For information how to install the Prep option to a standard OPTIMAS refer
appendix D.
Injection volumes Prep Mode

Ensure that the programmed injection volume does not
exceed the 50% of the loop volume, otherwise the
reproducibility and accuracy of the injection can not be
guaranteed.
6.4 Method programming mode
The autosampler offers nine programmable methods; depending on your needs, a

method may consist of:
Injection program Containing information on the type of the
injection with the flush volume, injection volume,
number of injections per vial and the analysis
time.
Wash program Containing information on the wash of the
OPTIMAS.
Mix program Containing information on the sample handling
of the sample prior to the injection.
Timed events program Containing information on the auxiliaries.
A method is any combination of the above and is saved as combination under its
method number.
To enter the method programming mode press [Methods].
40
Enter the method number; OPTIMAS will display the main screen of method
programming:
PROGRAMMING METHOD NUMBER: 1 (I W M T)

SELECT SECTION TO PROGRAM
<INJECTION> <WASH> <MIX> <TIMED EV.>
From this screen the various sections can be programmed after pressing the
corresponding function key.
Existing programs in this method are displayed between brackets on the first line:
 I for the injection program
 W for the wash program
 M for the mix program
 T for the timed events program.
When you have programmed the method, press [Escape] to leave the method
programming mode.
The methods are stored in battery backup memory.
6.4.1 <INJECTION>
Press <INJECTION> to enter the injection programming mode.
METHOD NUMBER: 1 SECTION: INJECTION

INJECTION MODE: PARTIAL LOOPFILL
<FULL> <PARTIAL> <PICK-UP> <NONE>
Select the mode of injection to be used or select <NONE> if no injection is required

in this method (see section 4 for an explanation of the injection principles).
<FULL> Full loop injection mode; see section 4.1
<PARTIAL> Partial loopfill injection mode; see section 4.2
<PICK-UP> µL Pick-up injection mode; see section 4.3
<NONE> No injection program in this method.
Depending on the injection mode you selected, the following parameters can be
entered:
Parameter: Range: Comment:
Flush volume 0-9999 µL The amount of sample used to fill
the needle plus tubing connection
to the injection valve (not the loop-
fill volume). Only for full loop and
partial loopfill injections.
41
Number of 1-9 Number of injections taken from

injections per vial the same vial.
Injection volume 0-2500 µL Only for partial loopfill and µL pick-
up injections
Analysis time Max.9:59:59 Analysis time starting at the
moment of switching the injection
valve from LOAD to INJECT.
Flush volumes smaller than twice the volume of the

needle plus tubing between needle and injection valve
may result in a decrease of performance!
The maximum programmable injection volumes are:
Full loop: Not programmable, is equal to the loop

volume but needs more sample to fill
the loop completely:
3 x loop volume for loops <= 100 µL
2 x loop volume for loops from 100 µL to
500 µL
1½ x loop volume for loops > 500 µL
Partial loopfill: 50% of the programmed loop volumes.
µL pick-up: Max. injection volume = ½ x loop
volume - 1½ x needle tubing
When all parameters have been programmed, return to the main method
programming screen by pressing [Escape].
6.4.2 <WASH>
Press <WASH> to enter the wash programming mode:
METHOD NUMBER: 1 SECTION: WASH

WASH BETWEEN: INJECTION
<INJECTION> <VIALS> <SERIES> <NONE>
42
The following parameters can be set for a wash method:

Parameter Range Comment
Wash between <INJECTION> Wash the needle and tubing directly
after every injection
<VIALS> Wash the needle and tubing directly
after every vial
<SERIES> Wash the needle and tubing directly
after every Series
<NONE> Remove the wash action from the
method.
No. syringe [0-9] Wash volume in number of syringe
volumes volumes
If the wash program is the only program used in a

method, program a wash between Series, otherwise
the Series with this method will not be executed!
6.4.3 <MIX>
Press <MIX> to enter the mix programming mode. A question will appear, to ask if
the mix program must be used in this method. Press <YES> to define the mix
program.
If <NO> is pressed the mix program is not used in this

method and the OPTIMAS returns to the previous screen.
The content of the mix program is NOT removed and will
be available the next time <YES> is pressed.
METHOD NUMBER: 1 SECTION: MIX LINE:01

END OF MIX
<DOWN> <UP> <INSERT> <DELETE>
The mix method may contain a maximum of 15 programmable lines (including the
line for END OF MIX).
 To scroll through the program steps use the <DOWN> and <UP> function keys.
 Press <DELETE> to delete the displayed step.
 Press [Enter] to edit the displayed step.
43
 Press <INSERT> to insert a step before the displayed step.
METHOD NUMBER: 1 SECTION: MIX STEP:01

SELECT ACTION:
<ADD> <MIX> <WAIT>
Three types of actions can be programmed:

<ADD> xxx µL from <SAMPLE> to <SAMPLE>
<REAG-A> <DESTINATION>
<REAG-B>
<WASH>
Aspirate a programmed amount of xxx µL (max.= the
syringe volume) from the sample vial, <SAMPLE>, a
reagent vial, <REAG-A> or <REAG-B> or from the wash
solvent bottle, <WASH>, and dispense it into the sample
vial, <SAMPLE>, or the destination vial,
<DESTINATION>.
To prevent cross contamination the autosampler will
aspirate an additional volume of 25% of the
programmed volume to flush the tubing and needle.
The aspirate and dispense speed depends on the
selected syringe and syringe speeds; programmed in
the System Settings, see section x.
If the mix is programmed the reagents and transport

solvent are positioned on the following places on the
tray:
Transport solvent: vial 85
Reagent A: vial 86
Reagent B: vial 87
Example: ADD 200 µL from Reagent A to Destination.

This will result in the following actions:
Aspirate an air segment of 5 µL, to separate the wash
solvent in the buffer tubing from Reagent A.
Aspirate 50 µL Reagent A; to flush the tubing and
needle.
Unload the syringe to the syringe-waste position.
Aspirate 200 µL Reagent A and dispense it to the
destination vial.
Rinse buffer tubing and needle with wash solvent.
<MIX> x times with xxx µL

Program a mix step: aspirate and dispense a number (x)
of times a programmed amount (xxx µL) from the
destination vial. If no destination vial is used the mix is
performed in the sample vial.
44
Example: MIX 3 times with 250 µL. This will result in the
following actions:
Aspirate an air segment of 50 µL, to separate the wash
solvent in the buffer tubing from solvent to be mixed.
Unload the syringe to the syringe-waste position.
Aspirate 250 µL the solvent and dispense it back into
the vial.
Repeat the last action two times.
Rinse buffer tubing and needle with wash solvent.
<WAIT> x:xx:xx
Wait a programmed period of time before continuing
with the next step (reaction time).
6.4.4 <TIMED EVENTS>

Press <TIMED EV.> to define a timed events program. The OPTIMAS will ask if
the timed events program must be used in this method; press <YES> to start
defining the timed events.
If <NO> is pressed the timed events program is not used

in this method and the OPTIMAS returns to the previous
screen. The content of the timed events program is NOT
removed and will be available the next time <YES> is
pressed.
METHOD NUMBER: 1 SECTION: TIMED EVENTS

1. AUX-1 ON AT TIME: 0:00:00
<AUX-1> <AUX-2> <END>
The timed events program is a list of 20 programmable events to control other

devices via the two auxiliaries (contact closures). Each event can be programmed
on a time base, which starts at the moment of injection.
Press [Clear] to remove the programmed time, the display will show the value:
NONE.
A complete list is shown below, to step through the list use the [ENTER] key or one
of the function keys to step immediately to the first line of a different section.
45
Timed events programming lines overview, as they appear on screen:

<AUX-1> 1. AUX-1 ON AT TIME: 0:00:00
1. AUX-1 OFF AT TIME: 0:00:00
2. AUX-1 ON AT TIME: 0:00:00
3. AUX-1 ON AT TIME: 0:00:00
4. AUX-1 ON AT TIME: 0:00:00
<AUX-2> 1. AUX-2 ON AT TIME: 0:00:00
2. AUX-2 ON AT TIME: 0:00:00
3. AUX-2 ON AT TIME: 0:00:00
4. AUX-2 ON AT TIME: 0:00:00
<END> END OF TIMED EVENTS AT: 0:00:00
If NO initial setpoint is programmed the lines with

additional setpoint changes will not be displayed.
If the END time exceeds the programmed analysis time,

this END time overrules the analysis time.
The next injection will start as soon as the timed events
program has finished!
It is possible to program events after the END time, but

these events are not executed during the RUN.
46
6.5 Series programming mode
In a Series or Run sequence, methods are linked to vials.
To enter the Series programming mode, press [Series]:
PROGRAMMING SERIES
SERIES NUMBER: 1
Enter a number [1-9] and press [Enter].
The first parameter of a Series is a method number. Depending on the content of

this method and on the System Settings, the OPTIMAS will display a number of
questions. Press [Enter] to step through the questions.
Each time a value has to be entered, key in a number [1-9] or press [CL] or 0 to
select NONE and press [Enter] to go to the next screen.
Method number [1-9] or NONE

The value <NONE> can be used for empty Series,
when programmed, the Series will be skipped
during the Run of the OPTIMAS.
This is an easy way to remove a Series or leave
the possibility to add Series during the Run, see
section 6.6.5 for programming during Run.
Should a method consist of only the wash and/or a
timed events program, no more questions will be
asked.
Use calibration vial Only available when use of calibration vials is
enabled in the System settings (see section 6.3)
and if the method does not contain a mix.
<YES>, enter: <NO>
- First calibration vial
- Last calibration vial
- No. of samples between
calibrations
Range of sample Enter:
vials - First sample vial
- Last sample vial
First destination vial Only available if the method contains a mix in
which destination vials are used.
47
Enter the position of the first destination vial.

The last destination vial is calculated on the basis
the the sample vial range defined earlier.
Figure 9: Run sequence with 3 calibration vials between every 5 vials (First calibration vial: 1, last
calibration vial: 3 and number of samples between calibrations: 5).
Sample vials and destination vials must be placed

sequentially.
Sample vial and destination vial ranges must not overlap.
When all parameters have been programmed, return to the Series programming
screen by pressing [Escape]. Another Series can now be programmed. If all Series
have been programmed, return to the Ready screen by pressing [Escape].
Series are not stored in battery backup memory. After

switching ON, the Series are empty (method: <NONE> ).
6.6 Run options
This section describes the options that can be used during a run.
6.6.1 [Start/Stop]
Use [Start/Stop] when the Ready screen is displayed to start a Run.
The OPTIMAS will ask for the Series number to start with and the last Series
number.
Enter each number and press [Enter] to go to the next screen.
48
EXECUTE SERIES 1
<START> <REMOTE>
The autosampler can be started directly by pressing <START>, or press

<REMOTE> to start remote control (see section 6.6.4).
If a Mix or µL Pick-up injection method is used in any

Series, make sure the transport liquid vials and/or reagent
vials are filled completely. If not, the autosampler might
aspirate air into the loop.
To return to the Ready screen without starting the OPTIMAS, press [Escape].
During the Run the OPTIMAS shows the following information:
STATUS: RUNNING TIME: 0:00:00

SERIES:01 SAMPLE:01 INJ: 1/1
METHOD:01 TRAY:--°C
AUX: 12
Explanation of the display:

 The first line shows the status and the elapsed analysis time.
 The second line shows the Series number, the sample vial number, the current
injection number, the total number of injections.
 The third line shows the method number of the current used method, this will
also be the method used when a priority sample is programmed (see section
6.6.2). When the tray cooling is installed the actual tray cooling temperature is
displayed; "- -" indicates the tray cooling is OFF.
 The fourth line shows information on the auxiliaries, when timed events are
programmed in a method. When the number is displayed behind "AUX:", this
indicates that the auxiliary is active.
Series are always executed in numerical order!

Empty Series will be skipped!
Press [Start/Stop] again to stop the Run. The OPTIMAS will execute a shutdown
sequence by removing all the sample from the buffer tubing and performing a
standard wash routine.
49
6.6.2 [Priority]
OPTIMAS offers the possibility to analyze an emergency sample without stopping
the current Run.
Press [Priority], to enter the priority-programming mode.
Enter the position of the priority sample.
PROGRAMMING PRIORITY
PRIORITY SAMPLE: 12
Return to the Run screen by pressing [Escape], the priority sample will be
processed after the current sample has finished with the method that is running at
the moment of entering the vial number.

p AUX: 12
To indicate the priority sample has been activated, a blinking "p" is displayed in the
Run screen.
As long as the "p" is blinking it is possible to change or
remove the priority sample.
The priority mode is only available during the execution

of a Series with a method, which does NOT contain a
Mix.
If calibration vials are used in the current Series, the

priority sample is included in the calibration interval!
6.6.3 [Hold Cont.]

Press [HOLD CONT.] to hold or continue the countdown of the analysis time. The
analysis times is extended with the time that Hold is active.
Press the button again to Continue the analysis time.
50
6.6.4 Remote control

After programming the range of Series to be Run, press <REMOTE> to start the
OPTIMAS in Remote Control mode. The OPTIMAS will now operate as slave of
another device and can be controlled with the Next Injection Input.
With the Next Injection Input OPTIMAS will start the next injection in the Series.
When the Series is completed, it starts the next injection from the next Series, until
all Series have been completed.
During the Run, OPTIMAS shows the following information:

r AUX: 12
A blinking "r" in the lower left corner of the Run screen indicates that the
autosampler is running in Remote Control mode.
After completion of all Series the OPTIMAS will display the following screen:
SERIES COMPLETED VIA REMOTE CONTROL
As long as this screen is displayed, it is not possible to

re-start the autosampler with a Next Injection Input.
To return to the Ready screen press [Escape]. From the Ready screen the
OPTIMAS can be re-started by pressing the [Start/Stop] key.
6.6.5 Programming during the run

During Run of the OPTIMAS it is possible to program Series and Methods:
Press [Series] or [Methods].
The programming possibilities are the same as described in sections 6.4 and 6.5.
51
If a Series or Method is changed, the new values are active

for Series after the current Series.
The current Series is NOT affected by changes.
It is not possible to add Series to the Run, but you can program empty Series
between the first and last Series in a Run. As long as the empty Series have not
been executed, they can be (re-) programmed. In this way Series with vials can be
added later to the Run.
52
7 Maintenance
For all maintenance procedures, open the cover of the autosampler.
You need not disconnect the autosampler from the

power source for any of the maintenance procedures.
7.1 Injection valve and rotor seal
The autosampler is equipped with a valve with quick connect mounting.
Execute the following steps to remove the injection valve:
1. Disconnect all tubing from the valve. Only the sample loop can stay in place.
2. Place the needle arm in the front position by pressing <MAINTENANCE>

<NDL HRZ> (see also section 6.2.2).
3. Remove the valve: push the pushers A backward to release the valve.
4. Pull the valve out of the instrument while turning the valve counter-clockwise.
Figure 10: Remove the valve
To remove the rotor seal:
1. Remove the screws from the stator part of the valve.
2. Gently open the valve and take out the rotor seal.
3. Clean and/or replace the seal.
53
4. Place the stator back on the rotor and fasten the screws.
To install the valve, proceed as follows:
1. Hold the valve for the mounting with the port 1 position facing to the top.
2. Push the valve into the mounting and turn the valve clockwise till it clicks in the
fitting.
3. Re-connect all tubing to the valve facing to the top.
4. Insure that the valve is in the INJECT position. Switching the OPTIMAS OFF
and ON can do this, all Series will be reset.
5. Perform a standard wash.
Figure 11: Install the valve
7.2 Needle and tubing
Two sizes of syringes are available for the dispenser: 250 and 1000 µL.
The 250 µL syringe is the standard syringe; combined with the standard 500 µL
buffer and the standard 100 µL sample loop, the following injection volume range is
available for the various injection modes:
 Full loop : 100 µL
 Partial loopfill: 1 - 50 µL
 µL pick-up : 1 - 27 µL
The maximum injection volumes are calculated with the following formulas:
 Full loop : injection volume =loop volume
 Partial loopfill: max. inj. volume =½ x of loop volume
 µL pick- up : max. inj. volume =½ x loop volume - 1½ × needle
volume
54
Full loop gives maximum possible reproducibility, but not

maximum accuracy, since loop volume is specified with an
accuracy of ± 10%.
Minimum sample loss = 230 µL (2 x loop overfill + flush
volume for needle).
Partial loopfill gives maximum accuracy (dependent on

syringe accuracy) plus reproducibility better than 0.5%
RSD for injection volumes > 10 µL
Minimum sample loss (Flush volume) = 30 µL
30 µL is the recommended minimum flush volume, smaller
flush volumes can be programmed, but will result in
decreasing performance.
µL Pick-up offers zero sample loss, maximum accuracy

(same as partial loop fill), but slightly less reproducibility:
RSD better than 1% for injection volumes > 10 µL.
5 µL of AIR is injected together with the sample, in case an
air segment is selected in the System Settings.
Sometimes other combinations of syringe, loop and/or buffer are advised:
Injection volumes smaller than 5 µL:

Partial loopfill Preferably install a 20 µL sample loop to avoid loss of
accuracy due to expansion of the loop content when
switching from inject to load position prior to sample
loading. Especially when working with high pressure (200
bar), this loss may be in the order of 0.1 - 0.5 µL for a 100
µL loop.
Note that the minimum sample loss in partial loop fill mode
is 30 µL (recommended minimum flush volume) for the first
injection and an additional 15 (always half the programmed
flush volume) for additional injections from the same vial. If
a wash between injections has been programmed, sample
loss is 30 µL for every injection. For zero sample loss
injections, use the µL-pick injection mode.
µL Pick-up Do not install a smaller sample loop!

The sample plug is transported into the loop, preceded by a
programmable air segment of 5 µL, with a plug of transport
liquid, which equals 2.5 times the programmed needle
tubing volume. The 20 µL loop is too small to guarantee
quantitative injection.
55
Injection volumes up to twice the standard:

With the standard 250 µL syringe, standard needle with tubing (15 µL) and
standard 500 µL buffer, but with a 200 µL sample loop, the maximum injection
volumes are:
Full loop: 200 µL Sample loss remains 230 µL since loops > 100
µL need only one loop volume overfill
Partial loopfill: 100 µL
µL Pick-up: 77 µL
Smaller volumes than 10 µL may be injected, but

reproducibility and accuracy may not be < 0.5% for partial
loop fill or < 1% for µL pick-up!
For volumes larger than 200 µL:

Install the 2000 µL buffer, install the appropriate sample loop size and the
appropriate syringe: Syringe volume > 2 x injection volume. Injection volumes
larger than 500 µL are possible, but the sample may contaminate the syringe.
Program sufficient wash after use!
Buffer tubing:
Syringe: Buffer tubing:
250 µL 500 µL
1000 µL 2000 µL
56
7.3 Sample needle
Figure 12: Sample needle
1. Ferrule
2. Nut
3. Tefzel tubing
4. Needle connection nut
5. Ferrule (PTFE)
6. Conical ring
7. Sample needle
8. Silicon air seal
9. Needle holder
10. Air needle
11. Vial sensor
12. Needle penetration depth adjustment screw
13. Air tubing to provide headspace pressure
57
7.3.1 Sample needle replacement

To replace the sample needle proceed as follows (see figure 12):
1. Place the sample needle in the front position, press <MAINTENANCE>, <NDL-
HOR>.
2. Loosen needle connection nut (4) that fixates the sample needle (7).
3. Loosen the nut (1) that connects the PTFE tubing (3) to the injection valve.
4. Remove sample needle by pulling it out of its fitting with the PTFE tubing that
usually sticks to the needle.
5. Put in a new needle assembly, watch the air seal (6) is around the needle.
6. Tighten the needle assembly with needle connection nut (4),
7. Connect loose end of needle connection tubing to port 4 of injection valve.

Do not tighten excessively; it may block the tubing!.
8. Place the sample needle back in the home position and leave the maintenance
mode, press <NDL HOME> and [Escape].
9. Check the needle penetration depth, see section and adjust if necessary.
10. Perform a wash routine to clean the new needle by pressing <WASH>.
7.3.2 Air needle replacement

To replace the air needle proceed as follows (see figure 12):
1. Remove sample needle as described in section 7.3.1.
2. Remove air needle (10) and install new air needle.
3. Re-install sample needle as described in section 7.3.1.
7.3.3 Sample needle height setting

To keep rest volume small, the sample needle should stop close to the bottom of
the sample vial. If the needle penetration depth needs adjustment, proceed as
follows (see figure 12):
1. Place an uncapped sample vial in position 2 of the tray.
2. Lower the sample needle in the vial:

a. Program a method with a full loop injection and an analysis time of at least
10 seconds.
58
b. Program a Series with the programmed method and vial 2 as first and last
vial.
c. Start the Series.
d. As soon as the sample needle has entered the vial, switch off the power
supply.
3. Loosen adjustment screw one turn (counterclockwise).
4. Use the needle connection nut (4) as a grip, adjust needle penetration depth.
Needle should stop not less than 1 mm above the bottom of a standard vial
and not less than 2 mm above the bottom of a conical vial.
5. Fasten adjustment screw (clockwise).
6. Switch on the power supply; the OPTIMAS will initialize and is ready for use.
7.4 Syringe
The autosampler is factory equipped with a 250 µL syringe, but can also be
equipped with a 1000 µL syringe.
Figure 13: Removing the syringe
To install another syringe proceed as follows:
Do not disconnect the power supply to the

autosampler, it is needed to move the syringe.
1. Press <MAINTENANCE> to get access to the maintenance function for moving

the syringe.
59
2. Move the syringe to the end position by pressing <SYR END>.
3. Unscrew the syringe from syringe valve.
4. Disconnect the plunger from the syringe drive.
5. Fill the new syringe with wash solvent, be sure most air bubbles are removed
from the syringe.
6. Connect the plunger of the filled syringe to the syringe drive and connect the
syringe at the syringe valve.
7. Screw the syringe firmly into the connector.
8. Remove the air from the syringe by pressing <SYR HOME>. The syringe will
move to its HOME position and dispense its contents to the syringe waste.
9. If some air remains in the syringe press <SYR END> again. The syringe is
filled with wash solvent.
10. Press <SYR HOME> to dispense the wash solvent to waste. If there is still air
in the syringe, repeat the previous action and tap the syringe gently as the
wash solvent is dispensed to the syringe waste.
11. Leave the maintenance screen by pressing [Escape] and press <WASH> to
perform a standard wash routine. All tubing connected to the syringe valve will
be refilled and flushed.
The autosampler is now ready for use.
60
8 Specifications
General
Sound pressure level LAeq <70 dB
Working temperature 10 - 40°C, indoor use only
Humidity 20 - 80% RH, indoor use only
Safety and EMC compatibility According to EC-directives
Installation class II
Pollution degree 2
Altitude up to 2000m
Dimensions (W x D x H) 300 mm x 500 mm x 340 mm
11.8". x 19.7" x 13.4"
Weight 19 kg (42 lb.)
21 kg (46 lb.) with cool option
Power requirements 115/230 VAC; ±15%; 50 Hz / 60 Hz; 250 VA
Fuses For 115 VAC: two 5.0 AT-fuses
(slow, ¼" x 1¼", UL/CSA)
For 230 VAC: two 2.5 AT-fuses
(slow, 5 x 20 mm, IEC 127)
Sampling
Sample capacity: Standard tray:
84 vials of 1.5 mL (Std) and
3 vials of 10 mL (LSV)
Vial dimensions (cap Maximum vial height: 47 mm
included): Minimum vial height: 32 mm
Loop volume 1 - 5000 µL
Dispenser syringe 250 µL (standard) or 1000 µL syringe

Vial detection Missing vial detection by vial sensor.
Headspace pressure Built-in compressor
Switching time injection valve Electrically <100 msec

Piercing precision needle ± 0.6 mm
Wash solvent External wash solvent bottle

Wetted parts in flow path SS316, PTFE, TEFZEL®, VESPEL®, Glass,
Teflon, PEEK
61
Analytical performance
Reproducibility RSD <= 0.3% for full loop injections
RSD <= 0.5% for partial loopfill injections,
injection volumes >10 µL
RSD <= 1.0% for µL pick-up injections,
injection volumes >10 µL
Memory effect < 0.05% with programmable needle wash
Programming
Methods 9 programmable methods
Injection method Injection mode:
Full loop injection
Partial loopfill injection
µL pick-up injection
Injection volume 1 µL - 5000 µL, with 1 µL increment
Full loop injection: max. 5000 µL
Partial loopfill injection: max. 2500 µL
µL pick-up injection: max. 2477 µL
Injections per vial max. 9 injections
Analysis time max. 9 hr 59 min 59 sec
Wash Programmable:
Wash between injections
Wash between vials
Wash between Series
Mix max. 15 programmable steps
Timed events Programmable:
4 x AUX-1 ON/OFF
4 x AUX-2 ON/OFF
Series 9 programmable Series
Priority sample Programmable
Communication
Outputs Inject marker (Relay & TTL)
2 Auxiliary outputs (Relay)
Alarm output (Relay)
Inputs Next injection input (TTL)
Freeze input (TTL)
Stop input (TTL)
Options (user-installable)
Prep option Including software, large volume tray, 2.5 mL
syringe and large bore Valco injection valve
with 10 mL loop.
Bio-compatible valve Special PEEK injection valve, quick connect
mounting.
62
Option (factory-installed)
Sample tray cooling Built-in Peltier cooling
Range: 4°C - 15°C
Accuracy: ± 2°C (at temperature sensor)
(Temperature at relative humidity of 80% and
ambient temperature of 30°C)
Cooling 20°C below ambient.
capacity:
63
64
9 Trouble shooting
This section offers basic troubleshooting information. Contact your service provider
if you come across a problem that you cannot solve.
65
9.1 Start up problems
66
9.2 Analytical problems
In case of analytical problems, determine whether the cause of the problem is the
autosampler, or the rest of the system.
Quick check!
Replace the valve by a manual injection valve to
discriminate between valve problems and other problems.
Perform some manual Full loop injections. If the results are
fine the fault has to be found in the autosampler, if not the
HPLC system should be checked.
Figure 14: Optimas flow path
In the diagrams on the next pages it is assumed that the

unit is working without Errors.
Please keep in mind that analytical 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 is changed.
67
9.2.1 No injection
68
9.2.2 Bad reproducibility
69
70
10 Installation
This section describes installation procedures for the Optimas autosampler.
Unpacking
Inspect the OPTIMAS for signs of damage. Damage that occurs to the OPTIMAS
as in transit, indicated by damaged containers, is the responsibility of the carrier
and should be reported to the carrier immediately. Shipping containers should be
inspected by the carrier if a claim is filed.
For contents of shipping containers refer to the packing list in the container.
Before using the OPTIMAS allow the instrument to

acclimatize for a period of 1 hour.
Store the shipping container and packaging materials of the OPTIMAS; we

recommend that you reuse the packaging materials and container if the OPTIMAS
must be shipped for service purposes.
Lift the OPTIMAS as shown in figure 15:
Figure 15: Lifting instruction
Lift the OPTIMAS with both hands under the instrument or with one hand under the
front while you hold the top rear of OPTIMAS with your other hand.
71
Do NOT lift the OPTIMAS at the front cover of the

autosampler.
Power connections
Before plugging in the power cable, check voltage setting of the OPTIMAS at the
input socket on the rear-panel. Make sure that the voltage setting is identical with
the voltage of your local power supply.
Understanding power surges

Power surges, line spikes, and transient energy sources
can impede instrument operation. Ensure that the
instrument's electrical supply is adequately protected
from these conditions and properly grounded.
If the indicated voltage is not correct select the proper voltage by removing,
inverting, and then re-entering the voltage selector-cartridge.
Check if the right fuses are installed, if not replace them with the right fuses.
For 115 VAC ±15%, use two 5 AT-fuses (slow).

For 230 VAC ±15%, use two 2.5 AT-fuses (slow).
All fuses must be UL listed and CSA certified!
RISK OF FIRE,
REPLACE FUSES AS MARKED.
Location
The OPTIMAS needs approximately 30 cm (12 inches) of bench space and one
power supply (230 VAC or 115 VAC, 50/60 Hz).
The best place to put your OPTIMAS is at the left-hand side of your HPLC system,
next to your HPLC pump and detector. From the right-hand side you have the
shortest connection to the injection valve of the OPTIMAS.
72
Keep the keyboard front cover closed during

operation.
Make that none of the ventilation holes are blocked.

Blockage of the ventilation holes can cause
malfunctioning of the OPTIMAS autosampler or even
damage the electronics inside.
Do not install the OPTIMAS in areas subject to

excessive dust, direct sunlight or shocks and do not
place it near any other source of heat, this will disturb
the cooling of the tray when installed.
Installation steps
Execute the following steps to install the OPTIMAS (if fluid connections have been
created already):
1. Place the OPTIMAS in the desired location and leave it there to adopt ambient
temperature for at least one hour.
2. Check OPTIMAS for visual damage; if this is the case, please contact your
distributor immediately.
3. Preferably, place the OPTIMAS autosampler on the left-hand side of your

HPLC system.
4. Remove the safety screw on the right-hand side of the front cover.
5. Check fuses and voltage settings on the rear side of the instrument.
6. Connect the power cable.
7. Switch on the OPTIMAS. The display shows the firmware revision number.
8. Connect the drain tubing to the wash outlet (figure 3, number 5).
9. Fill the wash solvent bottle with distilled water/ 2-Propanol (80/20 v/v%) or
mobile phase. Only use water/organic solvents; do not use any crystalline or
buffer solutions, as these may cause damage to the system. Degas wash
solvents to prevent air bubbles in the syringe.
10. Fill the wash solvent tubing, syringe and buffer tubing by washing the system 2
or 3 times: press <WASH> in the Ready screen.
11. Check if there are any air bubbles trapped in the syringe. Remove them by
gently tapping the syringe.
12. Connect your HPLC pump to port 1 of the injection valve.
13. Connect the column (or capillary) to port 6 of the injection valve.
14. Check for leakage and let the system equilibrate for at least 5 minutes.
73
System settings (see also section 6.3 of this manual).

These settings are factory-installed. To check the standard settings, proceed as
follows:
[System] Enter the System Settings
<GENERAL> Enter the General Settings
[0100] [Enter] Volume of installed loop: 100 µL.
[015] [Enter] Volume of tubing needle « valve:015 µL
<250> [Enter] Syringe volume: 250 µL
<NORMAL> [Enter] Set Syringe speed to Normal.
<YES> [Enter] Skip missing vials.
<YES> [Enter] Air segment: use air segment
<YES> [Enter] Headspace pressure: On
Press [Escape] twice to go back to the Ready screen.
10.1 Fluid connections
The OPTIMAS is factory installed with a 250 µL syringe, a 100 µL loop, a 500 µL
buffer tubing and a stainless steel sample needle.
A schematic presentation of all parts that have contact with any kind of fluid is
shown below. This sticker can be found on the inside of the front cover.
To obtain an optimal performance of the OPTIMAS, a correct combination of

syringe, loop and buffer tubing should be installed.
Figure 16: Fluid connections for HPLC
74
Overview of installed standard tubing:

Tubing Material and dimensions Label
Standard sample SS tubing; 70 mm x 0.65 mm OD x 0.25 15 µL
needle and tubing mm ID
Tefzel tubing; 155 mm x 1/16" OD x 0.25
mm ID
Buffer tubing from high- PTFE tubing; 640 mm x 1/16" OD x 1.0 500 µL
pressure valve to mm ID
syringe valve
Tubing syringe valve to PTFE tubing; 300 mm x 1/16" OD x 1.0
wash solvent bottle mm ID
Tubing syringe valve to PTFE tubing; 400 mm x 1/16" OD x 1.6
waste. mm ID
10.1.1 HPLC connections

To ensure reproducible injections the following connections to your HPLC system
should be made:
1. Connect the HPLC pump to port 1 of the injection valve.
2. Connect the HPLC column to port 6 of the injection valve.

The instrument has been flushed with Isopropanol: make sure that the mobile
phase of your HPLC system is miscible with Isopropanol. If not, start up with an
intermediate solvent as mobile phase (disconnect the HPLC column).
It is essential that the content of the sample loop is injected

in back flush onto the column, therefore: do not exchange
column and pump connections at the injection valve.
10.1.2 Waste tubing

Connect the drain tubing (in the shipkit of the OPTIMAS) to the right-hand
connector of the OPTIMAS and put it in a bottle. Through this drain all the liquid
dispensed to the wash position is removed.
Sample that is not injected will also be removed through

this tubing.
75
Syringe waste
Place the syringe waste tubing into a small bottle next to the OPTIMAS. If no
injection volumes are programmed that are larger than the buffer tubing can
contain, the syringe waste will only be wash solvent.
Condense water and leakage drain

Through the left-hand connector all leakage solvents and condensed water (if the
Peltier cool option is installed) is drained.
If the optional Peltier tray cooling is installed, it is advisable to connect this hose
connector (in the shipkit of the OPTIMAS) to a waste container on the floor.
Make sure that the drain and waste tubing are not twisted
to prevent an obstruction of the flow path.
10.1.3 Wash solvent and system rinse

Use a clean bottle for the wash solvent and place it at the left-hand side of the
OPTIMAS.
It is recommended to use a mixture of distilled water /

Isopropanol (80/20 v/v%) or mobile phase as wash
solvent. Before using the wash solvent, degas the solvent
with Helium or an ultrasonic bath.
Do not use salts or buffer solutions; crystals may block or
damage the system.
To fill the wash solvent tubing use the following procedure:
1. Put the wash solvent tubing in the filled wash solvent bottle.
2. Press <MAINTENANCE> to go to the OPTIMAS maintenance functions.
3. Fill the tubing using the OPTIMAS soft-function keys <SYR END> and <SYR
HOME>.
With <SYR END> a syringe volume of wash solvent is aspirated from the wash
solvent bottle and the wash solvent tubing is filled.
With <SYR HOME> the syringe contents will be dispensed to the syringe
waste.
4. Repeat this action until the wash solvent tubing and the syringe are completely
filled.
5. Press [Escape] to leave the maintenance screen.
76
After the wash solvent tubing and the syringe are filled, press <WASH> to perform
a standard wash routine. All tubing connected to the syringe valve will be rinsed
with wash solvent.
10.1.4 Syringe
The OPTIMAS is factory equipped with a 250 µL syringe, but can also be equipped
with a 1000 µL syringe. See section 7.4 for more information on how to replace the
syringe.
The OPTIMAS will give the best results if all air is removed
from the syringe.
Use <WASH> to remove air from the syringe, if this is not
satisfactory, see section 7.4 for more tips on removing air.
10.2 Preparation of vials
The OPTIMAS is standard equipped with a tray which can contain 84 standard
sample and three large volume vials for reagents and transport liquid.
Vials used should meet the following dimensions:

Maximum vial height, including cap: 47 mm
Minimum vial height, including cap: 32 mm
Maximum vial diameter:
Standard sample vial: 12 mm
Reagent / transport liquid vials: 22 mm
Large sample volume vials: 22 mm
The following section contains examples of Chromacol vials witch can be used in
the OPTIMAS. See appendix A, List of accessories, for the corresponding article
numbers for the vials, seals and caps.
10.2.1 Types of vials

The following vial types can be used in OPTIMAS:
 Chromacol standard sample vials
 Chromacol reagent and transport liquid vials
 Eppendorf micro centrifuge tubes
All types will be further described below.
77
Chromacol standard sample vials overview:
Figure 17: Standard sample vials
Figure 18: Conical vials with support
Figure 19: Plastic vials
78
To reduce the volume of standard vials, use a Chromacol insert (02–MTV, 02–
MTVWG or 03–MTV) in combination with the appropriate vial and support sleeve or
spring.
Figure 20: Standard vials with insert
It is possible to use the Chromacol 2.5-CV vials in the OPTIMAS.

When used, this vial type must be programmed in the System Settings, see section
6.3.
OPTIMAS uses two aspiration heights to aspirate liquid

from the 2.5-CV vials.
Therefore the vials need to be full, when OPTIMAS is
started.
When a vial is not full the OPTIMAS might aspirate air!
Chromacol reagent and transport liquid vial

The following Chromacol vial can be used for Reagents and transport liquid, this
vial can also be used in the optional LSV tray.
Figure 21: Reagent and transport liquid vial
79
Only use the following seal in combination with the 10-CV

vials: Chromacol: 20-ST101
together with one of the following caps:

Chromacol: 20-ACB (Blank aluminum cap)
20-MCB (Blank tinplate cap)
Do not use the 10-CV vials without being capped.

The OPTIMAS vial sensor will NOT detect uncapped vials!
OPTIMAS uses two aspiration heights to aspirate liquid from

the reagent and transport liquid vials.
Therefore the vials need to be full, when OPTIMAS is
started.
When a vial is not full the OPTIMAS might aspirate air!
Eppendorf micro centrifuge tubes

Eppendorf micro centrifuge tubes may be used instead of the standard vials.
To avoid cross-contamination prevent the air needle to make contact with the
sample.
Figure 22: Eppendorf micro centrifuge tubes with support sleeve
When using Eppendorf micro centrifuge tubes in the

OPTIMAS do not leave open spaces on the tray. The vial
detection system may not always detect a missing tube
because of the shape of the tubes.
80
10.2.2 Filling and sealing the vials

The standard vials, as well as the conical vials, can best be filled by means of a
narrow-end pipette to allow air to escape when filling the vial.
Do not fill vials to the edge to prevent that sample is forced into the air needle,
risking extra cross-contamination of samples and fouling of the needle pair.
It is important that the seal is airtight, to maintain a pressure on the vial for air
bubble prevention and to prevent evaporation of volatile samples.
When using uncapped vials, the performance of the

injections may not meet the specifications anymore
(precision).
Do not re-use a sample vial frequently without replacing
its cap or septum.
Check the seal after crimping: if the cap can be turned easily, the seal is not airtight
(re-adjust the hand–crimper).
10.2.3 Loading the sample tray

Place vial in the tray and link them to methods in the Series (see section 6.5).
To get access the tray can be rotated manually when OPTIMAS is not running.
10.3 Control I/O connections
OPTIMAS has standard two I/O connectors:

 Contact closures output connector (P1)
 TTL input connector (P2).
The manufacturer will not accept any liability for damages

directly or indirectly caused by connecting this machine to
instruments which do not meet relevant safety standard.
10.3.1 Contact closures output (P1)

Connector P1: Contact closure outputs
Pin Description: Pin no: Description:
no:
1 Inject marker 1 - Normally open 10 Alarm output - Normally open
81
2 Inject marker 1 - Common 11 Alarm output - Common

3 Inject marker 1 - Normally 12 Alarm output - Normally closed
closed
4 Auxiliary 1 - Normally open 13 Auxiliary 2 - Normally open
5 Auxiliary 1 - Common 14 Auxiliary 2 - Common
6 Auxiliary 1 - Normally closed 15 Auxiliary 2 - Normally closed
7 Inject marker 2 - Normally open
8 Inject marker 2 - Common
9 Inject marker 2 - Normally
closed
Vmax = 28 Vdc / Vac , Imax = 0.25 A
An Inject marker output (1 and 2) will be generated

when the injection valve switches from LOAD to
INJECT.
The Alarm Output will be activated whenever an error
occurs, see appendix B for a description of the error
codes of the OPTIMAS.
Figure 23: Contact closure output
10.3.2 TTL input (P2)

This connector is an active high or active low TTL input, user definable in the
System Settings. The Next Injection Input can be used when the OPTIMAS works
in Remote Control. The Freeze Input and Stop Input can be used to control the
OPTIMAS by other devices.
Connector P2: TTL inputs.

Pin no: Description:
1 Next Inject Input
2 Signal Ground
3 Freeze Input
4 Stop Input
5-9 Signal Ground
82
Figure 24: TTL input
Next Injection Input:

This input will start the next injection sequence when the OPTIMAS is started in
Remote Control. After finishing the injection sequence the OPTIMAS will wait for
the Next Injection Input.
From the Ready screen a Next Injection Input will start the latest programmed
Series. In this case the OPTIMAS will not wait for the Next Injection Input before
continuing with the next injection. The OPTIMAS will execute the complete Run as
if it was started with the [Start/Stop] key.
Freeze input:
The OPTIMAS will freeze the analysis time for the time this input is active. If the
Freeze Input is activated while the analysis time is not running, the OPTIMAS will
perform all programmed pre-injection sample handling (mix method and sample
loop). But the OPTIMAS will wait with injecting the sample until the Freeze Input is
no longer active.
Stop Input:
With this input the run of the OPTIMAS is immediately aborted and return to the
Ready screen. In case the OPTIMAS is in Remote Control, the Run of the
OPTIMAS is immediately aborted but the OPTIMAS remains in Remote Control
and can not be re-started with a Next Injection Input.
83
84
Appendix A. Spares and accessories
Article no. Description

0820.836 PREP option
0820.832 BIO kit
0820.316 Large capacity tray option, 96 position for 1.5 mL vials (12 mm OD)
0820.835 Large volume tray option, 24 position for 10 mL vials (22 mm OD),
with 1 mL syringe and LSV needle
0700.010 Flanged-tube fitting 1/8" (pck/5)
0700.011 Flanged-tube fitting 1/16" (pck/5)
0820.718 Air needle (incl. seal) OPTIMAS
0820.303 Sample needle OPTIMAS
0820.304 Serum sample needle OPTIMAS
0820.318 Silc/steel sample needle OPTIMAS
0820.711 Air needle for serum needle
0900.710 Buffer tubing 500 µL
0900.711 Buffer tubing 2000 µL
0900.712 Syringe waste tubing
0900.713 Syringe wash tubing
0830.811 Inject marker cable
0820.738 Injection valve, including loop and bayonet pins
0820.739 Injection valve PEEK with bayonet pins
0700.004 Valco Ferrule (pck/10)
0700.005 Valco Nut (pck/10)
3796.046 Valco loop 5 µL with fittings
3796.088 Valco loop 20 µL PEEK with fittings
3796.085 Valco loop 100 µL PEEK with fittings
3796.083 Valco fingertight nut PEEK
3796.084 Valco Ferrule PEEK
3796.045 Socket wrench for 1/16"nut
4400.250 250 µL syringe
4400.000 1000 µL syringe
0820.850 Wash bottle bracket for 2 x 250 mL wash bottles, incl wash bottles
0700.901 Preventive maintenance kit
85
86
Appendix B. Error codes
Injection valve
ERROR 211 Injection valve is not in a valid position.
ERROR 212 The injection valve did not switch within 1.5 seconds.
ERROR 213 The switching time of the injection valve exceeds 500
msec
Syringe dispenser unit

ERROR 221 The syringe valve did not switch.
ERROR 222 The syringe did not reach the home position within time.
ERROR 223 The syringe spindle did not make the correct number of
rotations.
ERROR 224 The spindle does not rotate.
ERROR 225 The syringe valve did not find a valid position.
Injection needle unit

ERROR 230 The sample needle arm did not reach or leave the
home position (vertical).
ERROR 231 The sample needle arm is in an invalid horizontal
position while moving down.
ERROR 232 The sample needle arm did not reach its destination
within a certain time (horizontal).
ERROR 233 Too many or not enough steps needed to reach
destination of horizontal needle movement.
ERROR 234 Sample needle arm not in vertical home position while
moving horizontally.
ERROR 235 The sample needle arm is at an invalid horizontal
position
ERROR 237 The sample tray is not at a valid position while moving
the sample needle arm down.
ERROR 239 Vial sensor sticks.
Tray unit
ERROR 251 Incorrect tray rotation
ERROR 252 No sample tray positioned in the OPTIMAS.
ERROR 253 The sample needle arm is not in the home position
while rotating the tray.
Vials
ERROR 260 Missing vial. Only available when Skip
Missing Vial is set to NO in the
System Settings (see section )
and during the execution of the
87
Mix of a method.
ERROR 262 Missing transport vial. Only available when Skip
Missing Vial is set to NO in the
System Settings (see section ).
ERROR 264 Missing vial for reagent A
ERROR 265 Missing vial for reagent B
ERROR 268 Missing destination vial.
ERROR 269 Not enough transport liquid available due to missing
transport vials.
ERROR 270 Not enough reagent A available.
ERROR 271 Not enough reagent B available.
Electronics
ERROR 275 Error occurred during initialization, the OPTIMAS
cannot start.
88
Appendix C. Test procedure
The OPTIMAS is factory tested for reproducibility, carry-over and mixing according
the following test procedure.
Analytical parameters:
Pump: Flow: mL/min
Eluent: distilled water
Detector: Wave length: 254 nm
Sample: Uracil in distilled water (50 ppm)
Figure 25: Configuration analytical system
Relative standard deviation (RSD%)

The formulas for calculating the Relative Standard Deviation (RSD%) are:
Reproducibility and carry-over test

Seven vials are placed in the OPTIMAS tray, vial 1 and vials 3 till 7 are filled with
sample (Uracil 50 ppm) and vial 2 is filled with eluent.
The wash solvent bottle is filled with 80% H2O / 20% Isopropanol.
89
The vials are run with the following method:

Injection
Injection mode: Partial loopfill
Flush volume: 30 µL
No of inj./vial: 3
Injection volume: 10 µL
Analysis time: 0:01:00
Wash
Wash between: None
An example chromatogram of the reproducibility test:
Peak Area
number
1 206342
2 204773
3 205425
4 -
5 -
6 -
7 205203
8 205566
9 205568
10 206156
11 206174
12 205956
13 206165
14 205765
14 205960
90
16 206296
17 205617
18 205619
19 205648
20 206141
21 207070
From the integration results the Relative Standard Deviation (RSD%) and the
carry-over can be calculated. For this example the results are:
RSD% = 0.25%
Carry over < 0.5%
Mix test
For the mix test one sample vial (50 ppm Uracil), one empty vial and the full
Reagent-A vial, filled with eluent, are placed in the OPTIMAS tray. A 1:10 dilution
of the sample will be made in the empty destination vial. After the dilution from both
vials (sample and destination) 10 µL will be injected. The wash solvent bottle is
filled with 80% H2O / 20 % Isopropanol.
The vials are run with the following methods:

Method 1:
Injection
No of inj./vial: 3
Wash
Wash between: None
Mix
1. ADD 180 µL REAG-A to DESTINATION
2. ADD 180 µL REAG-A to DESTINATION
3. ADD 40 µL SAMPLE to DESTINATION
4. MIX 3 times with 250 µL
5. END OF MIX METHOD
Method 2:
Injection
No of inj./vial: 3
91

Wash
Wash between: None
Program the following Series:

Series 1 Series 2
Method: 1 Method: 2
First vial: 01 First vial: 01
Last vial: 01 Last vial: 01
Destination vial: 02
An example chromatogram of the mixing test:
From the integration results the Relative Standard Deviation (RSD%) for the two
vials can be calculated.
Peak Area
number
1 38802
2 38968
3 39088
4 395258
5 394090
6 394471
For this example the results are:
RSD% (vial 1, 1:10 dilution) = 0.37%
RSD% (vial 2, sample) = 0.15%
1:10 dilution = 1 : 10.13
92
Appendix D. Prep option
By just choosing PREP in your system settings, you can use the OPTIMAS to inject
all of your sample into a Preparative LC system or in other areas where large
injection volumes are required. Also in combination with the PROSPEKT On-line
Solid Phase Extraction System, the OPTIMAS can be your ideal autosampler.
The combination of large sample vials (10 mL) a large sample volume needle and
a 2.5 mL syringe enable you to inject large volumes very reproducible with high
speeds and only 45 µL of sample loss. The installed large bore valve (0.75 mm)
with 10 mL sample loop enables you to inject from microliters to milliliters with the
same autosampler. Flow rates up to 200 mL per minute are no problem for the
OPTIMAS autosampler, used in Prep mode.
If the Prep option is factory-installed the part on installation can be skipped.

If the Prep option is bought as a kit, carry out the installation instructions.
Specifications
The Specifications of the Prep option differ in some ways from the standard
autosampler. The different specifications are listed below.
Sampling
Sample capacity LSV tray: 24 vials of 10 mL (LSV)
Loop volume Not programmable, injection volume
determines the aspirated sample volume
Dispenser syringe 2500 µL syringe
Analytical performance
Reproducibility RSD <= 1.0% for partial loopfill injections,
injection volumes >10 µL up to 50% of the
installed sample loop.
Memory effect < 0.1% with programmable needle wash
Programming
Methods 9 programmable methods
Injection method Partial loopfill only
Injection volume 1 µL – 19,999 µL, with 1 µL increment
Mix Not available
93
Option
Bio-compatible valve Not available
Installation
To install the Prep option on the standard autosampler perform the following
installation steps:
1. Replace the standard injection valve with the special Prep valve (VALCO #C2-
3006 SPHMI).
2. Replace standard sample needle, air needle and buffer tubing with the ones
supplied in the Prep kit. See section 7.3 for detailed information.
3. Connect tubing to the injection valve.
4. Replace the standard syringe with the 2500 µL syringe; see section 7.4 for
detailed information.
5. Install the 24 vials (LSV) tray.
6. Choose Prep Mode in the system settings, see section 6.3 .
Tubing of the Prep option:

Tubing Material and dimensions Label
LSV sample needle and SS tubing; 70 mm x 0.81 mm OD x 0.51 mm ID 45 µL
tubing. Tefzel tubing; 155 mm x 1/16" OD x 0.50 mm
ID
Buffer tubing from high- PTFE tubing; 2550 mm x 1/16" OD x 1.0 mm 2000 µL
pressure valve to syringe ID
valve.
Injections
The injection routine differs from the standard autosampler. The Prep version has
only the Partial loopfill injection routine, which differs from the partial loopfill
injection routine of the standard autosampler.
Therefore note the following for the best injection performance:
94
Ensure that the programmed injection volume does not exceed

the 50% of the loop volume, otherwise the reproducibility and
accuracy of the injection cannot be guaranteed.
The software does not check the injection volume with the
installed loop volume. It is therefore possible to program
injection volumes larger than 50% of the loop volume.
The preflush is no longer executed, expansion of the liquid in

the sample loop will remove sample from the sample tubing and
needle when switching from INJECT to LOAD. To compensate
for the volume of the needle and tubing, an additional amount of
the sample, equal to the programmed sample needle – tubing
volume (default 45 µL) is aspirated in the sample loop together
with the programmed sample volume.
95
96
Appendix E. Log book
Customer information:
Name of user
Company
Department
Address
Telephone
Fax
OPTIMAS information:
Serial number
Firmware revision
Purchase date
Installed options
Local Dealer
Service Engineer
Address
Telephone
Fax
E-mail
The following pages contain tables that can be used to

make your own OPTIMAS logbook.
Keep a record of your System Settings and programmed
methods. Therefore make copies of the following pages
(before filling in the forms).
97
System settings sample
General
Loop volume: µL
Needle volume: µL
Syringe volume:  250-µL syringe
 1000-µL syringe
Syringe speed:  Low
 Normal
 High
Skip missing vials:  Yes
 No
Air segment:  Yes
 No
Headspace pressure:  Yes
 No
Method
Use calibration vials:  Enabled
 Disabled
Tray
Tray type:  <84+3>, standard tray.
 <96>, tray with 96 standard vials
 <24>, large volume tray.
Vial type:  <STANDARD>, standard 1.8 mL vials.
(only when <84+3> or <96> is selected)  <2.5 mL>, 2.5 mL vials.
I/O configuration
Inject marker pulse length: sec.
Input edge next injection:  Falling
 Rising
Freeze input active:  Low
 High
Reset outputs after last Series:  Yes
 No
Prep Mode
Use Prep Mode:  Enabled
 Disabled
98
Method log sample
No:
 INJECTION  WASH
Injection Type:  Full loop Wash:  between injections
 Partial loopfill  between vials
 µL Pick-up  between Series
 None  None
Flush volume: µL Wash volume: X syringe volume
Injection / vial:
Injection µL
volume:
Analysis time:
 TIMED EVENTS
1. AUX-1 ON AT TIME:
1. AUX-1 OFF AT TIME:
END OF TIMED EVENTS AT:
 MIX
1.
2.
3.
4.
5.
99
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
100
Appendix F. Storage, packaging, recycling and disposal
If the autosampler needs to be stored for a long time, or if it must be shipped to a

different location, proceed as follows:
1. Thoroughly flush the system.
2. Disconnect and remove all tubing, except for the sample loop.
3. Switch off the autosampler.
4. In case of biohazard materials: remove and throw away the needle and other
flow path materials, in accordance with an approved waste disposal program.
5. Use the original packaging materials to package the autosampler.
6. Fill in a health & safety form for the instrument and include the form in the
package.
7. Dispatch the package; make sure any tax/import/export requirements are met.
Contact your local distributor if you need more information.
Decontamination
The instrument shall be decontaminated before decommissioning and all local
regulations shall be followed with regard to scrapping of the equipment.
General instructions for disposal

When taking the instrument out of service, the different materials must be
separated and recycled according to national and local environmental regulations.
Hazardous substances
The instrument contains hazardous substances. Contact the manufacturer for more
detailed information.
Disposal of electrical components

Waste of electrical and electronic equipment must not be disposed as unsorted
municipal waste and must be collected separately. Please contact an authorized
representative of the manufacturer for information concerning the decommissioning
of equipment.
101
102
E
Emergency sample • 50
Index
Example - 10 uL Partial loopfill injection from
vial number 1 • 26
Example - 3 x 10 µL pickup injections • 27
[
Example - a 1
[Hold Cont.] • 50
10 dilution + 10 µL partial loopfill injection •
[Priority] • 50
29
[Start/Stop] • 48
F
<
Filling and sealing the vials • 81
<GENERAL> • 36
Fluid connections • 74
<INJECTION> • 41
Full loop injections • 17
<IO> • 39
H
<MAINTENANCE> • 34
Hold analysis time • 50
<METHOD> • 38
HPLC connections • 75
<MIX> • 43
I
<PREP> • 39
Identification of parts • 11
<SERVICE> • 35
Injection principles • 17
<TEMP> • 34
Injection valve and rotor seal • 53
<TIMED EVENTS> • 45
Injections • 94
<TRAY> • 38
Installation • 71, 94
<UTILS> • 35
Introduction • 9
<WASH> • 33, 42
K
µ
Keyboard and display • 14
µL pickup injections • 22
L
A
Loading the sample tray • 81
About this manual • 5
M
Air needle replacement • 58
Maintenance • 53
Analytical problems • 67
Method log sample • 99
Appendix A. Spares and accessories • 85
Method programming mode • 40
Appendix B. Error codes • 87
N
Appendix C. Test procedure • 89
Needle and tubing • 54
Appendix D. Prep option • 93
No injection • 68
Appendix E. Log book • 97
P
Appendix F. Storage, packaging, recycling
Partial loopfill injections • 19
and disposal • 101
Pictorials • 5
B
Preparation of vials • 77
Bad reproducibility • 69
Priority sample • 50
C
Programming during the run • 51
Contact closures output (P1) • 81
Programming overview • 31
Continue analysis time • 50
Programming reference • 31
Control I/O connections • 81
R
Ready screen options • 33
103
Remote control • 51
Rotor seal • 53
Run options • 48
S
Safety instructions • 6
Sample needle • 57
Sample needle height setting • 58
Sample needle replacement • 58
Series programming mode • 47
Specifications • 61, 93
Start up problems • 66
Syringe • 59, 77
System settings programming • 36
System settings sample • 98
T
Trouble shooting • 65
TTL input (P2) • 82
Types of vials • 77
Typographical conventions • 8
U
Using the OPTIMAS • 25
W
Wash solvent and system rinse • 76
Waste tubing • 75
104
Spark Holland B.V.
P.O. Box 388
7800 AJ Emmen
The Netherlands
Spark Holland -
Main office and factory:
P. de Keyserstraat 8
7825 VE Emmen
The Netherlands
Phone: +31 591 631700
Fax: +31 591 630035
E-mail: sales@sparkholland.com

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May 2014

BETTER SAMPLE CARE

Spark Holland B.V.

P +31 591 631 700

1 About this manual 5

5 Using the OPTIMAS 25

6.5 Series programming mode 47

Appendix A. Spares and accessories 85

Appendix B. Error codes 87

Appendix C. Test procedure 89

Appendix D. Prep option 93

Appendix E. Log book 97

Appendix F. Storage, packaging, recycling and disposal 101

1 About this manual

The following pictorials are used in this manual:

The danger sign warns about a hazard. It calls

The warning sign denotes a hazard. It calls attention

The caution sign denotes a hazard. It calls attention to

The attention sign signals relevant information. Read

The note sign signals additional information. It

This symbol indicates that the waste of electrical and

1.2 Safety instructions

Adhere to standard quality control procedures and the following equipment

1.3 Typographical conventions

function keys in pointed brackets, just like in the

PASA™, a robust injection concept

Figure 1: The PASA™ injection concept

Side-port needle: a good point!

Integrated sample cooling option

Reagent addition made simple

Service friendly design

OPTIMAS is a standalone autosampler of robust design. This section describes

Proceed as follows for access to the sampling compartment:

2. Carefully lift the cover (see figure 2).

3. Rest the cover on the top of the autosampler.

Figure 2: Access to sampling compartment

The following parts are important for use of the autosampler:

Figure 3: Main parts of the autosampler

4. Position for transport solvent and reagent vials

5. Wash and waste drain

6. Condensed water and leakage drain

7. Tray fastening knob

10. Display and keyboard (on cover)

The back of the autosampler offers the following:

Figure 4: Back of the autosampler

2. I/O connector (inputs)

3. Fan (only when tray cooling option is installed)

4. Fuse box and voltage selector

7. I/O connector (outputs)

3.1 Keyboard and display

The OPTIMAS is controlled by way of a keyboard and a display:

Figure 5: Display and keyboard

2. Function keys (see below for more information)

3. Numerical keypad (see below for explanation)

4. Start/Stop button (see below for explanation)

5. Hold/Continue button (see below for explanation)

6. Control keys (see below for explanation)

Explanation of keyboard keys:

[SYSTEM] Enter the System Settings programming mode. In

A unique sequence of valve switching and syringe dispenser controlled sample

Three different methods for injection can be selected:

Full loop: The sample loop is completely (quantitatively) filled with

4.1 Full loop injections