Autosampler 3950 Manual V1509A

Autosampler 3950 Manual V1509A
Autosampler 3950
Manual
V1509A
3
Table of contents
Note:
For your own safety, read the manual and always observe
the warnings and safety information on the device and in
the manual!
Table of contents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Local area network and automatic configuration . . . . . . . . . . . . . . . . . . . . . . . . 7
Laboratory use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Where is it prohibited to use the device or system? . . . . . . . . . . . . . . . . . . . . . . 7
Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Laboratory regulations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Solvents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Flushing solution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
PEEK connections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Protective measures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Power supply and mains connection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Target group . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Operating the device or device system . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
To what should the user pay particular 
attention? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
What expertise should users have to safely
operate a HPLC device or device system? . . . . . . . . . . . . . . . . . . . . . . . . . . 11
Symbols and labels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Packaging and transport . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Packaging material and shipping boxes . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Scope of supply . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Biocompatible version . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Preparative version . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Checking the scope of supply . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Space requirements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
13
14
14
14
14
Installation site . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
Standard . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Optional . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Biocompatible version . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Preparative version . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Startup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
15
15
15
15
15
Front view of the device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
Front view of device with sample compartment . . . . . . . . . . . . . . . . . . . . . 17
Rear view of the device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
V1509A
4
PASA™ loop injection principle. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Injection modes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Full loop injections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
Air segment with Full loop injections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
Partial loopfill injections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
Air segment with Partial loopfill injections . . . . . . . . . . . . . . . . . . . . . . . . . . 24
Microliter pick-up injection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
Air segment with μL Pickup injections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
μL Pickup injections with 84+3 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
Air segment with μL Pickup injections 84+3 . . . . . . . . . . . . . . . . . . . . . . . . 29
84+3 tray . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
μL Pickup method parameters for the 84+3 tray . . . . . . . . . . . . . . . . . . . . . . . 31
Programming details for the 84 + 3 injection mode . . . . . . . . . . . . . . . . . . 32
Air needles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
Standard air needle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
Choosing the correct air needle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
Calculation example for air needle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Handling the sample vials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
Mixing and diluting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
Example: Add . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
Example: Mix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
Sample positions in mix method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
Processing in columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
Processing in rows . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
Mix method parameters for the 84+3 tray . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
Programming details for the 84 + 3 mix method . . . . . . . . . . . . . . . . . . . . 40
Capillary and tube connections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
Connections on the injection valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
Syringe connections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
Tube guide for flushing solution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
Connecting the drainage tubing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
Connecting the autosampler to other devices . . . . . . . . . . . . . . . . . . . . 45
Controlling the autosampler with chromatography software . . . . . . . . . . . . . . 45
Checking and configuring the parameters of the autosampler . . . . . . . . . . 45
Configuration window of ClarityChrom® . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Autosampler device software . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
System flushing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
System flushing with Smartline Autosampler 3950 Service Manager . . . . . . . 46
I/O connection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Defining the TTL Inputs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
Defining the closed-contact output . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
Configuration of I/O connection (9-pin) . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
5
Device test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
Test intervals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
Devices and components for test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
1. Reproducibility of sample volume . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
Standard setting of autosampler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Method parameters of pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Method parameters of autosampler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Method parameters of UV detector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Configuring repeat runs of autosampler . . . . . . . . . . . . . . . . . . . . . . . . . . .
Starting repeat runs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Analyzing the individual chromatograms . . . . . . . . . . . . . . . . . . . . . . . . . .
Formula for determining the arithmetic
average . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
2. Sample carryover . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
50
50
50
50
50
50
51
Creating a sequence with 6 lines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Analyzing the individual chromatograms . . . . . . . . . . . . . . . . . . . . . . . . . .
Formula for calculating sample carryover . . . . . . . . . . . . . . . . . . . . . . . . . .
3. Linearity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
51
52
52
52
51
51
Analyzing the individual chromatograms . . . . . . . . . . . . . . . . . . . . . . . . . . 52
Formula for determining the correlation coefficient . . . . . . . . . . . . . . . . . . 53
4. Mixture test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
Creating a sequence with 2 lines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Positioning vials for dilution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Analyzing the individual chromatograms . . . . . . . . . . . . . . . . . . . . . . . . . .
Archiving . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
53
53
54
54
Test Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
Operation Qualification (OQ) with ChromGate® . . . . . . . . . . . . . . . . . . . . . . 56
Operation Qualification Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
Maintenance and care . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
KNAUER Technical Support . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
Maintenance contract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
What maintenance tasks can users perform on the device? . . . . . . . . . . . . . . . 58
Leaks in the capillary screw fittings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
Exchanging the fuses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
Exchanging the injection valve and rotor seal . . . . . . . . . . . . . . . . . . . . . . . . . 59
Removing the injection valve and rotor seal . . . . . . . . . . . . . . . . . . . . . . . .
Installing the rotor seal and injection valve . . . . . . . . . . . . . . . . . . . . . . . . .
System flushing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Exchanging the sample loop . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
60
61
61
61
Exchanging the sample needle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
Exchanging the air needle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
Exchanging the syringe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
Exchanging the syringe plunger or plunger tip . . . . . . . . . . . . . . . . . . . . . . 65
Exchanging the syringe valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
Cleaning and caring for the device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
Environmental protection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
Disposal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
6
Decontamination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
Device errors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
Checking the valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
Software fault . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
Analytical errors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
System messages from ChromGate® . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
Technical data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
Standard version . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
Preparative version . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85
Delivery program . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86
Devices and accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86
Spare parts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86
Accessories 84+3 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
Legal information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88
Warranty conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88
Declaration of conformity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89
Abbreviations and terminology. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90
Table of Figures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
7
Overview
Overview
Smartline
Autosampler 3950 HPLC
The Smartline Autosampler 3950 has been developed to meet
the demands of a modern analytical laboratory. It is robust and
compact, easy to install and to operate, and designed for high
throughput rates. For optimal results, sample cooling is possible.
The autosampler is generally installed as a core element in
a Smartline HPLC system.
Local area network and automatic
configuration
Remote control
Normally the autosampler is controlled by means of the
chromatography software, via a local network (LAN).
Automatic configuration
The autosampler connected to the local area network (LAN) is
automatically detected by the chromatography software.
Device status
When used in a local area network (LAN), the system status of
the autosampler can be checked using the chromatography
software.
Laboratory use
 Biochemistry analyses
 Chiral analyses
 Food analyses
 Pharmaceutical analyses
 Environmental analyses
Where is it prohibited to use the
device or system?
DANGER!
Risk of explosion! Never use the device in 
potentially explosive atmospheres without
appropriate protective equipment and 
approval by a notified body!
Inform the technical support department 
of the manufacturer!
8
Safety
Safety
When the door of the autosampler is open, the syringe speed is
reduced automatically. The auto-injection system can cause stab
injuries when handled inappropriately!
Use of autosampler
 Always operate the autosampler with its front door closed!
DANGER!
Risk of stab injuries! Never operate the autosampler when the front paneling is not in place!
Laboratory regulations
Adherence to
laboratory regulations
 Observe national and international regulations pertaining to
laboratory work!
 Good Laboratory Practice (GLP) of the American Food &
Drug Administration
 For development of methods and validation of devices:
Protocol for the Adoption of Analytical Methods in the
Clinical Chemistry Laboratory, American Journal of Medical
Technology, 44, 1, pages 30–37 (1978)
 Accident prevention regulations published by the accident
insurance companies for laboratory work
Solvents
Suitable solvents
Solvents suitable for use in HPLC:
 Acetone
 Acetonitrile
 Benzene
 Chloroform
 Acetic acid (10–50%), at 25 °C (77 ºF)
 Ethyl acetate
 Ethanol
 Hexane/heptane
 Isopropanol
 Methanol
 Phosphoric acid
 Toluol
 Water
Note:
Even small quantities of other substances, such as additives, modifiers, or salts can influence the durability of
the materials. The list of selected solvents was compiled based on research in the pertinent literature and
is only a recommendation by the manufacturer. In the
event of doubt, contact the technical support of the
manufacturer.
9
Solvent tray
Toxicity
Safety
To avoid damage from leaks, always place solvent bottles in
a solvent tray on the device.
Organic solvents are toxic above a certain concentration.
Ensure that work areas are always well-ventilated! Wear
protective gloves and safety glasses when working on the
device!
Flammability
Organic solvents are highly flammable. Since capillaries can
detach from their screw fittings and allow solvent to escape,
it is prohibited to have any open flames near the analytical 
system!
Self-ignition
point
Only use solvents that have a self-ignition point higher than
150 °C (302 ˚F) under normal ambient conditions!
Unsuitable solvents
The following solvents can attack the components of the pump
and are therefore not suitable:
 Mineral and organic acids (except in buffer solutions)
 Bases (except in buffer solutions)
 Liquids containing particles
Suitable to only
a limited extent
The following solvents are suitable to only a limited extent
for use in the pump:
 Methylene chloride
 Tetrahydrofuran (THF)
 Dimethyl sulfoxide (DMSO)
 Slightly volatile solvents
 Fluorinated hydrocarbons
Leaks and
clogged capillaries
 Regularly check for leaks and clogged capillaries – test back
pressure without column!
Flushing solution
Note:
Do not use a salt or buffer solution for flushing!
PEEK connections
 Disposable PEEK fittings
 One-piece disposable polyetheretherketone fittings
(PEEK) for easier installation of flexible, thin capillaries
 Tightening torque of the PEEK screw:
Tightened by hand (approx. 0.5 Nm)
10
Safety
Protective measures
1. You are only permitted to perform the maintenance tasks
described in this manual.
2. All other maintenance tasks are to be performed exclusively
by the manufacturer or a company authorized by the
manufacturer.
Without exception, the following applies to all maintenance
tasks that can be performed by the user:
1. Switch off the device and pull the power plug!
2. Never open the device! High voltage poses a life-threatening
risk!
Power supply and mains connection
The device is intended for use with AC power networks of 
100–240 V. The supplied power cable is to be used to connect
the device to the mains supply.
Note:
To disconnect the device from the mains power, 
disconnect the power plug on the rear of the device.
Fuses
If the fuses blow repeatedly, consult with KNAUER Technical
Support for replacements and help in identifying the cause.
Target group
Operating the device or device system
The device can be operated by means of the chromatography
software at the workstation, the touch screen or the membrane
keyboard at the device.
To what should the user pay particular 
attention?
To make your HPLC or UHPLC separations as efficient as possible, pay close attention to the following:
Avoiding additional
dead volumes
1. Once they have been used, never re-use capillaries in other
areas of the HPLC or UHPLC system.
2. Only use a given PEEK fitting for one specific port and never
re-use it for other ports. Always install new PEEK fittings on
each separate port.
Using special columns
 Use special columns and follow the manufacturer's instructions on caring for the columns!
Checking for clogged
capillaries
 Regularly check for clogged capillaries – test back pressure
without column!
11
Using filtered solvents
Safety
1. Use ultra-pure, filtered solvents – Gradient grade – for the
HPLC or UHPLC.
2. Filtration of substances to be analyzed.
3. Use of inline filters.
Devices are to be
opened by the technical
support department
only
Note:
Only allow the technical support department of the
manufacturer or a company authorized by the manufacturer to open the devices for maintenance and
repair work.
What expertise should users have to safely
operate a HPLC device or device system?
 Completed degree as chemical laboratory technician or comparable vocational training
 Fundamental knowledge of liquid chromatography
 Participation in an installation of the system performed by the
manufacturer or a company authorized by the manufacturer,
or suitable training on the system and chromatography software
 Basic knowledge of Microsoft Windows®
Knowledge regarding substances that are suitable only
to a limited extent for use in liquid chromatography
12
Symbols and labels
Symbols and labels
Explanations of symbols and labels on the autosampler
Symbol
Explanation
CE (Conformité Européenne) mark for
equipment that complies with the
pertinent EU directives and comes
with a declaration of conformity from
the manufacturer.
For your own safety, read the manual
and always observe the warnings and
safety information on the device and
in the manual!
13
Installation
Installation
Packaging and transport
At the factory, the device is carefully packed for safe transport.
Checking for signs of
damage during
transport
Check the device for signs of damage that occurred during
transport. If the shipment is incomplete or damaged, inform the
manufacturing factory within three workdays. Also inform the
freight carrier about transport damage.
Note:
Without packaging, the autosampler weighs approx.
19 kg (21 kg with cooling option). Therefore, the
autosampler should be transported and set up by 
two people.
Packaging material and shipping boxes
The device is held in place and protected by foam inserts at the
top and bottom. Please keep the transport box and the foam
inserts.
Removing the
packaging material
Remove device
from packaging
 Remove the foam insert on the top of the device.
 Grip the device at its sides near the front and lift it out of the
packaging.
Scope of supply
 Autosampler with injection valve and 500 μl syringe
Accessories
 Device manual
 CD: Smartline 3950 Autosampler Service Manager
 PTFE tube, 300 cm
 Silicon drainage tube, 200 cm
 2x sample plates for 48 vials
 5x microtiter plate, 96 wells
 2x 2.5 A fuses
 1x flushing solution bottle
 Cables
 Power supply cable
 Network cable
 I/O interface cable, 9-pin
14
Installation
Biocompatible version
 Inert sample needle with glass or titanium coating
 PEEK injection valve
Preparative version
 Syringe 2500 μl
 Sample loop 10 ml
 Preparative injection valve
Checking the scope of supply
1. Check whether the supplied devices and accessories are
complete.
2. If anything is missing, consult with KNAUER Technical
Support.
KNAUER Technical Support Hotline
European hotline
E-mail:
Languages: German and English
Available by telephone: 8 am to 5 pm (CET)
Phone:
+49–(0)30–809727–0
Fax:
+49–(0)30–8015010
[email protected]
Space requirements
 Side clearance to other devices:
 If there is a device on one side, min. clearance of 5 cm.
 If there are devices on both sides, min. clearance of
10 cm.
 If cooling fan is present at rear of device, minimum clearance
of 30 cm.
Installation site
Ambient conditions of
the installation site
 Air humidity: below 90% (non-condensing)
 Temperature range: 4–40 °C; 39.2–104 °F
 Set up the autosampler so that it is protected against
exposure to direct sunlight.
15
Features
Features
Standard
 Can be flexibly equipped with microtiter plates or standard
sample plates
 Maximum sample capacity 2x 384 or 96 standard
autosampler vials
 Injection cycle <60 s, incl. cleaning
 Full loop or partial loop injection, as well as μl pick-up mode
 Sample priority function
 Quick-exchange injection valve
 High-resolution syringe controller
Optional
 Sample cooling
Biocompatible version
 Inert sample needle with glass or titanium coating
 PEEK injection valve
Preparative version
 Syringe 2500 μl
 Sample loop 10 ml
 Preparative injection valve
Startup
Note:
Before starting up the autosampler for the first time,
wait approximately one hour until the temperature of
the device has adapted to the ambient temperature.
Front view of the device
The door of the autosampler can be pushed horizontally into 
the device. The front paneling can be completely removed. 
To replace the sample plate, remove the cooler cover.
DANGER!
Risk of stab injuries! Never operate the autosampler while the door is open!
16
Opening the door
Startup
 Lift the door from the side and push it into the interior.
Legend
A LED status indicator
A
Fig. 1
Removing the
front panel
Push the door into the interior
 Press both black buttons on either side (A) of the housing at
the same time and remove the front paneling.
A
Fig. 2
Removing the
cooler cover
Removing the front panel
 Pull out the cooler cover in the direction of the marked
arrows.
Fig. 3
Removing the cooler cover
17
Startup
Front view of device with sample compartment
Legend
A Injection valve
B Syringe
A
C Needle guide
B
C
D
D Collecting container
E Flushing solution
bottle
E
F
F Cooler cover for
sample plates
G Tube connector for
waste liquid
G
H
H Tube connector for
condensed water
Fig. 4
Front view of device with sample compartment
Rear view of the device
The following components, connections and warnings can be
found at the rear of the device:
Legend
A LAN connection
A BC
DE
B I/O connection 
(9-pin)
C Warning A
F
G
H
D Power switch
E Warning B
F CE mark
G Fuse box
I
H Mains connection
I Cooling fan
J
J Serial number 
and year of manufacture 
of device
Fig. 5
Rear view of the autosampler
18
PASA™ loop injection principle
PASA™ loop injection principle
The autosampler uses loop injection with pressure assistance
(Pressure Assisted Sample Aspiration PASA™) as a selectable
option. PASA™ injection has the following features:
 Samples do not have to be degassed.
 No air bubbles in sample loop.
 No clogging or contamination of sample needle.
 Precise control of syringe movement.
Legend
A Buffer tube
A
B
B Capillary to pump
C
C Sample loop
D
E
D Capillary to column
E Syringe
F Connector for compressed air
F
G Air needle
G
H Sample needle
I Sample vial
H
I
Fig. 6
PASA™ loop injection principle
Injection modes
Four different injection modes are available:
 Full loop injection
 Partial loop injection
 Microliter pick-up injection
 Microliter pick-up 84+3 injection
Full loop
injection
In full loop injection mode, the sample loop is completely filled
with the sample. The maximum reproducibility but not the
maximum precision is achieved, because the size of the sample
loop may have a deviation of ±10 %. The maximum injection
volume equals the loop volume. The sample loop is filled with
a multiple of the loop volume:
3 x loop volume for loops up to 100 μl
2 x loop volume for loops from 100 μl to 500 μl
1.5 x loop volume for loops of more than 500 μl
19
PASA™ loop injection principle
The sample loss per injection is the sum of the x-time overfilling
of the sample injection and the specific flush volume of the
employed needle.
Partial loop
injection
In partial loop injection mode, the sample loop is only partially
filled with the sample. This ensures the highest precision of the
sample volume with minimal loss of sample. The maximum
injection volume equals 50 % of the loop volume. The sample
loss per injection equals the specific flush volume of the
employed needle.
Microliter pick-up
injection
In microliter pick-up injection mode, the sample loop is filled
with a very small amount of sample that is transported by
washing solution. This ensures the highest precision of the
sample volume without loss of sample. The maximum sample
volume is 50 % of the loop volume, minus 1.5-times the needle
volume.
Microliter pick-up
injection 84 + 3
If the 84 + 3 tray is selected for the microliter pick-up injection,
the sample is transported by a separate transport liquid instead
of the washing solution. The consumption of transport liquid
depends on the needle volume and corresponds to 2.5-times
the needle volume for the segments before and after the
sample.
The autosampler uses a system of two telescopic needles, one
that pierces through the cap of the sample vial – the air needle,
– and one that extracts the sample – the sample needle.
Using a syringe, the sample is aspirated through the two needles
out of the sample vial while under pressure and into the sample
loop. To prevent the syringe from becoming contaminated,
a buffer tube is situated between the syringe and the valve.
Using washing solution, sample residue is removed from the
sample needle and buffer tube.
20
PASA™ loop injection principle
Full loop injections
The sample loop is completely filled with sample. This type of
injection results in extremely good reproducibility.
Full loop injection
Functional schematic
Fig. 7
Full loop: Initial situation
Explanation
1. The initial situation:
the injection valve is in
INJECT position. The
sample needle with air
needle has entered the
well or vial. Headspace
pressure, applied
through the 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 well/vial to fill
the sample line with
sample and remove
wash solvent.
Fig. 8
Full loop: The needle and
sample lines are flushed
3. The injection valve
switches to LOAD
position, placing a
distinct sample plug at
the inlet of the sample
loop.
Fig. 9
Full loop: Injection valve
switches to LOAD
position
21
PASA™ loop injection principle
Functional schematic
Fig. 10
Fig. 11
Full loop: Sample loop is
filled completely
Full loop: Injection valve
switches to INJECT
position
Explanation
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.
3 x loop volume for
loop 100 μL
2 x loop volume for
loops 100 μL - 500 μL
1.5 x loop volume for
loop 500 μL
5. The injection valve
switches to 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.
Note:
A wash routine is
performed after
each injection.
Air segment with Full loop injections
An air segment of 5 μL 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.
With a standard needle, the flush volumes must be a minimum
of 30 μL for injections with air segment, and 35 μL for injections
without air segment. If samples are highly viscous it may be
necessary to program larger flush volumes and reduce the
syringe speed for better performance.
Fig. 12
Full loop injection with air segment (A), 
without air segment (B)
22
PASA™ loop injection principle
Partial loopfill injections
This type of injection results in maximum accuracy.The
switching sequence for a partial loopfill injection is:
Patial loop injection
Functional schematic
Fig. 13
Partial loop: Initial
situation
Explanation
1. The initial situation:
the injection valve is in
the INJECT position.
The sample needle
with air needle has
entered the vial/well.
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/well to fill
the sample line with
sample and remove
wash solvent.
Fig. 14
Partial loop: The sample
line is filled with sample
3. The injection valve
switches to LOAD,
placing a distinct
sample plug at the
inlet of the sample
loop.
Fig. 15
Partial loop: Injection
valve switches to LOAD
position
23
PASA™ loop injection principle
Functional schematic
Explanation
4. The programmed
injection volume is
now aspirated into the
sample loop.
Fig. 16
Partial loop: Sample
loop is filled partially
5. The injection valve
switches to INJECT.
The sample loop is
now part of the HPLC
mobile phase flow
path: sample is
transported to the
column. The analysis
starts.
Fig. 17
Partial loop: Injection
valve switches to INJECT
position
If an injection from the same vial and no wash routine is
programmed, the next injection sequence will start with a flush
of 50% of the programmed flush volume. Otherwise, it will start
with a flush of the programmed flush volume. If the withdrawal
of sample for the next injection exceeds 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.
24
PASA™ loop injection principle
Air segment with Partial loopfill injections
An air segment can be used to reduce the amount of flush
volume. The air segment is at the front of the flush volume and
will not be injected.
With a standard needle, the flush volumes must be a minimum
of 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.
Fig. 18
Partial loop injection with air segment (A), 
without air segment (B)
25
PASA™ loop injection principle
Microliter pick-up injection
In this injection mode sample is transported into the sample
loop by washing liquid. This type of injection results in
maximum accuracy without sample loss. If you select the 84+3
tray with μL pickup, the three 10 mL vials are automatically used
for transport. The Needle Wash/Transport position in that case
will only be used for the needle wash. That means that a
different routine will be followed with first transport. Refer to the
section on 84+3 tray (see "μL Pickup method parameters for the
84+3 tray" on page 31) for more information.
The switching sequence for μL pickup injections is:
Microliter pick-up
injection
Functional schematic
Explanation
1. In the initial situation,
the injection valve is in
INJECT position. The
sample needle has
entered the transport
position. The transport
container is filled with
wash solvent.
Fig. 19
Microliter pick-up:
Washing liquid is
aspirated
2. Washing liquid is
aspirated into needle
and buffer tubing to
place a distinct sample
plug at the inlet of the
sample loop.Please
note that the washing
liquid needs to be
compatible with
eluent.
3. The injection valve
switches to LOAD
position. A transport
plug of washing
solution is aspirated
into the sample loop.
Fig. 20
Microliter pick-up:
Injection valve switches
to LOAD position
26
PASA™ loop injection principle
Functional schematic
Explanation
4. The needle moves
from the transport
position to the sample
vial/well.
Fig. 21
Microliter pick-up: The
needle moves to sample
vial/well
5. The programmed
injection volume is
aspirated from the
sample vial/well.
Fig. 22
Microliter pick-up:
Injection volume of
sample is aspirated
6. The sample needle
moves back to the
washing position. A
second transport plug
of washing solution is
aspirated. The sample
is transported into the
sample loop.
Fig. 23
Microliter pick-up:
Sample is transported
into sample loop
7. The injection valve
switches to INJECT.
The sample loop is
now part of the HPLC
mobile phase flow
path: sample is
transported to the
column. The analysis
timer starts.
Fig. 24
Microliter pick-up:
Sample is transported to
column
27
PASA™ loop injection principle
Air segment with μL Pickup injections
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.
In this injection mode:
 The air segment at the front of the sample plug is injected
into the HPLC system
 No headspace pressure can be applied on vials/wells in this
mode to avoid sample errors due to air expansion during
exchange from the sample vial/well to the transport position.
Fig. 25
Microliter pick-up injection with air segment (A), 
without air segment (B)
28
PASA™ loop injection principle
μL Pickup injections with 84+3
If you select the 84+3 tray with μL pickup, the three 10 mL vials
are automatically used for transport. The Needle Wash position
in that case will only be used for the needle wash. This results in
the following injection routine:
Microliter pick-up
injection with 84+3
Functional schematic
Explanation
1. At the start of the
routine, the needle is
still at the Needle
Wash position. The
valve is in INJECT
position.
Fig. 26
Microliter pick-up 84+3:
Initial situation
2. The syringe dispenser
aspirates a transport
plug from the
transport vial position
to fill the sample line
with transport liquid
and remove wash
solvents.
Fig. 27
Microliter pick-up 84+3:
Sample line is filled with
transport liquid
3. The injection valve
switches to LOAD
position.The
programmed injection
volume is aspirated
from the sample vial/
well.
Fig. 28
Microliter pick-up 84+3:
The sample is aspirated
29
PASA™ loop injection principle
Functional schematic
Explanation
4. The sample needle
moves back to the
transport vial position.
A second transport
plug is aspirated. The
sample is transported
into the sample loop.
Fig. 29
Microliter pick-up 84+3:
Sample is transported
into sample loop
5. The valve switches to
INJECT. The sample
loop is now part of the
HPLC mobile phase
flow path: sample is
transported to the
column. The analysis
timer starts.
Fig. 30
Microliter pick-up 84+3:
Injection valve switches
to INJECT position.
Air segment with μL Pickup injections 84+3
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.
In this injection mode:
 The air segment at the front of the sample plug is injected
into the HPLC system
 No headspace pressure can be applied on vials in this mode
to avoid sample errors due to air expansion during exchange
from the sample vial to the transport position.
30
PASA™ loop injection principle
84+3 tray
A 84+3 tray is available for the Alias autosampler. The tray is
placed over both tray positions in the autosampler.
Note:
Always place the tray with position 87 in the farmost
right corner in the sampling compartment.
The tray may contain a maximum of 84 vials of 1.5 mL + 3 vials
of 10 mL. The tray has been designed in such a manner that the
caps of the 1.5 mL and of the 10 mL vials are at the same height
to make sure that the same injection needle can be used for
both vial types:
Fig. 31
Height 84+3 tray
That means that the sample needle height can only be
programmed for the 1.5 mL vials. For the 10 mL vials two fixed
needle height levels are available, depending on the transport
liquid levels.
Fig. 32
Needle height and fluid levels
31
PASA™ loop injection principle
Please note that the processing order for this tray is fixed in
numerical order if you program a series of samples:
Fig. 33
Processing order
However, if you create sequence tables using one-line-one
sample (1 sample in each run), the sample processing order can
of course be programmed randomly.
μL Pickup method parameters for
the 84+3 tray
To indicate that you wish to use the 84+3 tray, change the tray
settings in the control software.
Note:
If you select the 84+3 tray with μL pickup, the three 
10 mL vials are automatically used for transport. The
Needle Wash position in that case will only be used for
the needle wash.
Sample positions are defined as follows for the 84+3 tray:
First sample position to be processed:
Vial positions 1 - 84
Last sample position to be processed:
Vial positions 1 - 84
First destination position to be
processed:
Vial positions 1 - 84
Transport positions:
Vial positions 85 - 87
Fig. 34
Transport/Reagent positions on 84+3 tray
The range of samples is indicated by the parameters First Sample
to Last Sample. Transport positions are fixed (vials 85, 86, 87).
32
PASA™ loop injection principle
Programming details for the 84 + 3 injection
mode
 The transport vial position can be programmed. Valid
positions are 85, 86 and 87. Position 85 is the default
transport vial position.The system calculates the transport
volume used. Make sure the vial contains at last 8000 μL
whenever starting the system.
 Whenever a run is started, the transport fluid levels are not
reset. In the transport vial, the needle stops at a high level in
the vial, to prevent the air needle from being polluted.
 When a series of samples or one-line-one-sample is
processed, the autosampler keeps track of the remaining
transport volume during the run and moves deeper in the
transport vial when transport volume level is lower than 4000
μL. When the amount of transport liquid reaches 0 μL, the
autosampler reports error 369 (not enough transport liquid).
 The transport volume levels are reset by reprogramming the
mode.
 The needle will not automatically move to a different
transport vial. You will have to program a different transport
position if you want to draw transport liquid from a different
transport vial position for further runs.
33
Air needles
Air needles
Six different lengths of air needles from 50 to 80 mm are
available for the autosampler. The needle holder allows you to
further adjust the needle height by 6 mm.
Standard air needle
The standard air needle is 62 mm long and can be used for a
broad range of deep and shallow sample plates.
When 10 ml sample vials are used, the needle penetrates deeply
into the sample vial. If this is not filled to more than 60%, 
the needle can be used in the typical manner. The same applies
to using deep microtiter plates.
For non-standard settings, use the corresponding needle types.
Legend
A 10 ml sample vial
A
B
B 1.5 ml sample vial
Fig. 35
Standard air needle with 10 and 1.5 ml sample vial
Note:
The PASA™ loop injection principle is not suitable for
shallow microtiter plates. The function of the air needle
is only ensured when it pierces through the closure.
Legend
A Deep Greiner
microtiter plate with
closure
A
B
B Shallow Greiner
microtiter plate
Fig. 36
Standard air needle with Greiner microtiter plates
34
Air needles
Choosing the correct air needle
 To choose the correct air needle, take the following
dimensions into consideration.
Ht = height of sample
plate
Dw = hole depth
Cd = thickness of
closure
Nh = set needle height
Ac = distance of the 
air needle tip to the
closure (min. 2 mm)
? = excess length
Fig. 37
Calculating the correct air needle
Condition:
Ht - Dw = 2 to 6 mm.
Protrusion length of the sample needle:
Ht - Cd - Nh - Ac = ?
 Choose the correct needle type on the basis of the protrusion
length:
Air needle type
Protrusion length
50 mm, yellow
34–40 mm
56 mm, red
28–34 mm
62 mm, white 
(standard needle)
22–28 mm
68 mm, blue
16–22 mm
74 mm, green
10–16 mm
80 mm, black
4–10 mm
35
Air needles
Legend
A 10 ml sample vials,
50 mm air needle
A
B
B 1.5 ml sample vials,
62 mm air needle
Fig. 38
Selected air needles with sample vials
Legend
A Deep Greiner
microtiter plate 
with closure, 
56 mm air needle
A
B
B Shallow Greiner
microtiter plate,
80 mm air needle
Fig. 39
Selected air needles with Greiner microtiter plates
Calculation example for air needle
Starting point:
 Autosampler with standard setting for needle height.
 Depth of Greiner microtiter plate with closure.
Dimensions:
Ht = 41.4 mm
Dw = 37.8 mm
Cd = 3.8 mm
Nh = 6.0 mm (standard)
Ac = 2.0 mm (minimum)
36
Air needles
Condition:
Ht - Dw = between 2 and 6 mm.
Ht - Dw = 41.4 mm - 37.8 mm = 3.6 mm. Condition has been met.
Ht - Cd - Nh - Ac = Protrusion length of the sample needle
41.4 mm - 3.8 mm - 6.0 mm - 2.0 mm = 29.6 mm
Air needle type
Protrusion length
56 mm, red
28–34 mm
An air needle length of 56 mm is required.
Handling the sample vials
When handling the sample vials, observe the following:
 Fill the sample vials using a pipette to allow air to escape.
 To prevent the sample from contaminating the air needle, 
do not fill the sample vials to the very top.
 Do not use sample vials that are unclosed.
 Only use air-tight closure seals to prevent air bubbles from
forming and volatile components from evaporating.
 Do not use sample vials with hard closures that the injection
needle cannot pierce.
37
Mixing and diluting
Mixing and diluting
A mix method can be programmed for the autosampler to mix
or thin the sample fluid.
 Configure the mix method and syringe speeds using the
chromatography software.
 A maximum of 15 steps can be programmed for a mix
method.
Three types of actions are possible:
1. Add
2. Mix
3. Wait
Add
When adding, the defined volume is aspirated from either the
sample vial, the vial with reagent A or reagent B or flushing fluid
and then dispensed into the destination vial.
Note:
To prevent carryover, the autosampler removes 125%
of the given volume from the corresponding sample
vial and uses the additional 25% to flush the tube and
needle.
Mix
With the Mix command, the contents of a specific sample vial
are mixed by aspirating and dispensing the defined volume 
n times. If a destination vial has not been defined, mixing is
performed in the current sample vial.
Wait
With the Wait command, the system waits until the
programmed delay time has elapsed before executing the next
line of the program.
Example: Add
The ADD 200 μl from Reagent A to Destination command triggers
the following steps:
1. An air segment of 5 μl is aspirated to separate the flushing
solution in the buffer tube from reagent A.
2. 50 μl of Reagent A are aspirated to flush the tube and needle.
3. Syringe is emptied into the waste container through the
drainage tube.
4. 200 μl of Reagent A are aspirated and then dispensed into
the destination vial.
5. Tube and needle are flushed with flushing solution.
Example: Mix
If this is preceded by an ADD … to Destination command,
mixing is performed in the destination vial. If this is preceded by
an ADD … to Sample command, mixing is performed in the
sample vial.
38
Mixing and diluting
The MIX 3 times with 250 μl command triggers the following
steps:
1. An air segment of 50 μl is aspirated to separate the flushing
solution in the buffer tube from the sample solution to be
mixed.
2. Syringe is emptied into the waste container through the
drainage tube.
3. 250 μl solution are aspirated and dispensed into the same
sample vial.
4. Step 3 is repeated twice.
5. Tube and needle are flushed with flushing solution.
Sample positions in mix method
When configuring a mix method, the positions of the sample
vials depend on whether the sample plates are to be processed
in rows or columns.
Processing in columns
When column processing is used, the following positions are
possible for the sample, destination, reagent A and reagent B:
Fig. 40
Sample positions in columns
39
Mixing and diluting
Processing in rows
When row processing is used, the following positions are
possible for the sample, destination, reagent A and reagent B:
Fig. 41
Sample positions in rows
40
Mixing and diluting
Mix method parameters for the 84+3
tray
To indicate that you wish to use the 84+3 tray, change the tray
settings in the control software.
Sample positions are defined as follows for the 84+3 tray:
First sample position to be processed:
Vial positions 1 - 84
Last sample position to be processed:
Vial positions 1 - 84
First destination position to be
processed:
Vial positions 1 - 84
Reagent positions:
Vial positions 85 - 87
Fig. 42
Transport/Reagent positions on 84+3 tray
The range of samples is indicated by the parameters “First
Sample to Last Sample”. The same range can be used for
“Destination Vials”. Reagent positions are fixed (vials 85, 86,
87).
Programming details for the 84 + 3 mix method
 The transport vial position can be programmed. Valid
positions are 85, 86 and 87. Position 85 is the default
transport vial position.The system calculates the transport
volume used. Make sure the vial contains at last 8000 μL
whenever starting the system.
 Whenever a run is started, the reagent fluid levels are not
reset. In the reagent vial, the needle stops at a high level in
the vial, to prevent the air needle from being polluted.
 When a series of samples or one-line-one-sample is
processed, the autosampler keeps track of the remaining
reagent volume during the run and moves deeper in the
41
Mixing and diluting
reagent vial when reagent volume level is lower than 4000
μL. When the amount of reagent liquid reached 0 μL, the
autosampler reports error 370 (not enough reagent).
 The reagent volume levels are reset by reprogramming the
mode.
 Reagent is used from the programmed reagent position only.
The needle will not automatically move to a different reagent
vial. If you want to use reagent from one of the other two
vials, you will have to program that setting.
42
Capillary and tube connections
Capillary and tube connections
Note:
Flush the system before connecting the column.
The schematic diagram inside the autosampler shows the
correct connections.
Fig. 43
Connection schematic for capillaries and tubing
Connections on the injection valve
Explanation
Component
A Stainless steel capillary
to pump
A
B Sample loop
C Plastic capillary to
syringe
D Plastic capillary to injection needle
E Sample loop
F Stainless steel capillary
to column
B
Fig. 44
C
D
E F
Connections on the
injection valve
43
Capillary and tube connections
Syringe connections
Explanation
Component
A For waste tube
AB C
B For buffer tube
C For washing solution
tube
Fig. 45
Syringe connections
Tube guide for flushing solution
Note:
Legend
A Tube guide for flushing solution
Use the tube guide in the collecting container for the
flushing solution to prevent the needle unit from
moving horizontally.
A
B
B Hole in collecting
container (top view)
Fig. 46
Tube guide for flushing solution tube
Connecting the drainage tubing
The waste drainage removes all flushing fluids and non-injected
sample solutions.
 Connect the condensed water and drainage tubing to the
front of the device.
 Connect the adapter and insert the end of the tube into
a container on the floor.
Note:
Make sure that the drainage tubing is not pinched so
that the liquid can flow away.
44
Capillary and tube connections
Legend
A Tube for waste liquid
A
B
C
B Tube for condensed
water
C Adapter for drainage tube
Fig. 47
Connecting the drainage tubing
45
Connecting the autosampler to other devices
Connecting the autosampler to other
devices
Controlling the autosampler with
chromatography software
The autosampler is controlled directly using chromatography
software, e.g. ChromGate® and ClarityChrom® from KNAUER.
 Using the LAN connection at the rear of the device, connect
the autosampler to the network.
Checking and configuring the parameters of
the autosampler
You can configure the autosampler parameters using the
chromatography software, e.g. ClarityChrom®:
1. Select autosampler in LAN.
2. Set the syringe volume to either 250 ml (default) or 500 ml.
3. Enter the serial number of the autosampler.
4. Select the cooling option if the autosampler is equipped wih
sample tray cooling.
Configuration window of ClarityChrom®
Legend
A Serial number
AB C
D
E
B Device detection in
local network
C Manual search for
device in network
D Volume of sample
loop
E Volume of syringe
Fig. 48
Autosampler configuration with ClarityChrom®
46
Connecting the autosampler to other devices
Autosampler device software
A software CD with Smartline Autosampler 3950 Service Manager
is included with the autosampler. Using this software, you can
check and control the device. 
These settings can also be made with the chromatography
software, so that it is not absolutely necessary to install the
Service Manager.
System flushing
The system should be flushed before the column is connected.
System flushing can be controlled using the chromatography
software or Smartline Autosampler 3950 Service Manager.
Note:
KNAUER recommends using a mixture of water and
isopropanol (80%/20%) or the mobile phase as the
flushing solution.
The following steps are explained for when the Service Manager
is used.
System flushing with Smartline Autosampler
3950 Service Manager
1. Install Smartline Autosampler 3950 Service Manager.
2. Fill the flushing solution into a solvent bottle and degas it
using helium or an ultrasonic bath.
3. Insert the flushing solution tube into the solvent bottle.
4. Select the Alias Direct Control menu.
5. In the Syringe field, click End. One syringe volume is aspirated
into the syringe through the flushing solution tube.
6. In the Syringe field, click Home. The syringe content is
emptied into the drainage tube.
7. Repeat step 5 and 6 until the syringe and the flushing
solution tube have been filled completely.
8. In the Initial wash field, click Start. All tubes that are
connected to the syringe are flushed.
9. In the Initial wash field, click Stop.
10. Click Close to exit the Direct Control window.
47
Practical tip!
Connecting the autosampler to other devices
Perform additional flushing to remove all of the air from the
syringe.
Fig. 49
System flushing with the Service Manager software
I/O connection
By default, the autosampler has an I/O connection that supports
TTL inputs (low-active) and a closed-contact output. Devices
without LAN connection that require a trigger signal for the
injection can be connected by means of the I/O connection.
The TTL inputs allow you to control the autosampler using other
devices and are defined using the instrument methods of the
chromatography software.
Note:
Only connect the autosampler to devices that fulfill the
required safety standards!
48
Connecting the autosampler to other devices
Defining the TTL Inputs
 Next Injection Input: The injection sequence is started. 
After ending the injection sequence, the autosampler waits
for the next start signal.
 Freeze Input: The analysis time is paused. The autosampler
runs the configured program up to the filling of the sample
loop. The injection is not performed until the input is
deactivated.
 Stop Input: Immediately halts the analysis.
Defining the closed-contact output
 Inject Marker: The closed-contact output is activated when
the injection valve switches from LOAD to INJECT.
 Alarm: The closed-contact output is activated when there is
an autosampler fault.
 Auxiliary: -
Configuration of I/O connection (9-pin)
Explanation
Cable color
1. Output,
start injection
Red in three-conductor cable
2. Output,
start injection
Black in three-conductor cable
3. Input 1,
programmable input
for stopping
injections (lowactive)
Red in four-conductor cable
4. Input 2,
programmable input
for stopping
injections (lowactive)
Black in four-conductor cable
5. Not occupied.
-
6. Output
Brown in three-conductor cable.
7. Not occupied.
-
8. Ground, for inputs 1
and 2
Orange in four-conductor cable
9. Ground, for inputs 1
and 2
Brown in four-conductor cable
49
Device test
Device test
The reproducibility of the sample volume is a critical factor for
maintaining high-quality analysis results.
 Test the autosampler features with the chromatography
software on a regular basis.
 If the device test determines that the autosampler does not
fulfill the requirements, mark the device as defective and do
not continue using it.
 Do not re-use the autosampler until it has been repaired and/
or serviced.
Test intervals
Run the device test at the following time intervals:
 Average use of 1 to 5 days/week: device test every 6 months
 Average use of more than 5 days/week or 24 hours/day:
device test every 3 months
 Operation with buffer solutions or other salt solutions: device
test every 3 months
Devices and components for test
 Smartline Autosampler 3950, standard version with 100 μl
sample loop and 500 μl syringe
 HPLC pump, 1 ml/min flow rate
 UV detector (with a data rate of 50 Hz if possible, else 10 Hz,
flow cell: 10 mm path length)
 Chromatography software
 Eluent: 90% water, 10% methanol (HPLC quality)
 Test solution (sample):
 a: 50 ppm uracil dissolved in water (HPLC quality)
 b: 250 ppm uracil dissolved in water (HPLC quality)
 Flushing solution:
 80% water, 20% isopropanol (HPLC quality)
 Alternative: 80% water, 20% methanol (HPLC quality)
 Restriction capillary:
 Inner diameter 0.25 mm
 Length 200 cm
Note:
Degas the eluent to prevent malfunctions caused by
the presence of air bubbles.
50
Device test
1. Reproducibility of sample volume
The variation coefficient must not exceed 0.5%.
 Configure the HPLC pump, UV detector and autosampler
using the chromatography software
 Inject 10 μl of test solution a (50 ppm uracil, dissolved in
water)
Standard setting of autosampler
 Loop volume: 100 μl
 Tubing volume: 15 μl
 Syringe volume: 500 μl
Method parameters of pump
 Flow: 1 ml/min
 Time: 1 min
Method parameters of autosampler
 Injection method: Partial loopfill
 Syringe speed: normal
 Flush volume: 30 μl, (40 μl for microtiter plates)
 Needle wash: active 2 times
 Air segment: yes
 Headspace pressure: yes
 Injections/vial: 9 (7 for microtiter plates)
 Inj. volume: 2 μl
 Vial position: 1A1. Start the single run with
Method parameters of UV detector
 Wavelength: 254 nm
 Sampling rate: 50 Hz if possible, otherwise 10 Hz
 Time: 0.5 min
Configuring repeat runs of autosampler
 Injections/vial: 9x (7x for microtiter plates)
 Inj. volume: 10 μl
Starting repeat runs
1. Put a vial with at least 500 μl test solution on position 1A1 of
the sample plate.
2. Start the repeat runs.
51
Device test
Analyzing the individual chromatograms
1. Calculate the average of the measuring values of the 
Peak areas.
2. Calculate the variation coefficient VK1.
3. Enter the results into the Test Report form.
Formula for determining the arithmetic
average
 Formula for determining the standard deviation (i = 1-9):
 Formula for determining the variation coefficient:
2. Sample carryover
The percentage of sample carryover must not exceed 0.3%.
 Alternately inject 10 μl of test solution b (250 ppm uracil,
dissolved in water) followed by eluent.
 Position of sample vial: 1A1
 Position of eluent: 1A2
 Injection volume: 10 μl
Creating a sequence with 6 lines
Note:
For microtiter plates, select 6 consecutive positions
that are alternately to be filled with test solution and
eluent.
 Test solution: Position 1A1
 Eluent: Position 1A2
 Injection volume: 10 μl
 Repeats: 1
52
Device test
Analyzing the individual chromatograms
1. Calculate the average of the measuring values of the 
Peak areas.
2. Put the average of the eluent injection in relation to the
average of the test solution injection.
3. Enter the results into the Test Report form.
Formula for calculating sample carryover
3. Linearity
To determine the linearity, the correlation coefficient of the
regression lines is determined from the measured values for the
Peak areas and the injection volume.
The correlation coefficient must not exceed 0.998%.
 Inject 10, 20, 30, 40 and 50 μl of test solution b (250 ppm
uracil, dissolved in water) respectively.
 Position of sample vial: 1A1
 Injection volume: 10 μl, 20 μl, 30 μl, 40 μl, 50 μl
 Repetition: 3
Note:
Fill consecutive positions on the microtiter plates with
test solution.
Analyzing the individual chromatograms
1. Calculate the correlation coefficient r of the regression lines
from the measured values for the Peak areas and the injection
volume.
2. Enter the results into the Test Report form.
53
Device test
Formula for determining the correlation
coefficient
yi = Y value of measured value i (injection volume)
xi = X value of measured value i (peak area)
= Arithmetic average of Y across all n measured values
= Arithmetic average of X across all n measured values
n = Number of measuring value pairs
4. Mixture test
To create a mixing method, follow the instructions in the
manual of the chromatography software.
 The variation coefficient VK2 must not exceed 0.5%.
 The variation coefficient VK3 for dilution must not exceed 
0.5%.
 The dilution factor F10 has to be within the range of 
9.85 < x < 10.25.
 Test solution: 10 μl (50 ppm uracil, dissolved in water)
 For injecting the dilution, go to Mix methods and create
a mixing method in which 40 μl of the test solution is mixed
with 360 μl eluent.
 Dilution: 10 μl, 5 ppm uracil, dissolved in deionized water
 Inject 10 μl test solution and und 10 μl dilution three times
each.
Creating a sequence with 2 lines
 Injection volume: 10 μl
 Repeats: 3
Positioning vials for dilution
Note:
Observe the plate assignments for the vials (sample,
reagent A, reagent B, destination).
 To position the vials on the sample plates, select the Columns
option in the chromatography software.
 Put a vial with test solution (sample) at position 2A1 of the
sample plate.
54
Device test
 Put an empty vial (destination) at position 2A5 of the sample
plate.
 Put a vial with eluent (reagent A) at position 1A1 of the
sample plate.
Analyzing the individual chromatograms
1. Calculate the average of the measuring values of the 
Peak areas for the test solution and the dilution.
2. Calculate the variation coefficient VK2.
3. Calculate the variation coefficient VK3.
4. Calculate the dilution factor F10 from the ratio of the
averages of the Peak areas of the test solution and the
dilution.
5. Enter the results into the Test Report form.
Archiving
 Enter all test results into the Test Report form.
 Enter the serial number, date of the test, date of the next test
and name of the tester.
 File the Test Report form in the device logbook.
55
Device test
Test Report
Module
Autosampler
Smartline 3950
Standard
Sample cooling
Biocompatible
version
Preparative
version
Serial number
No.
Test
Setting
Specification
1
Reproducibility
 Inject 10 μl test
solution nine times.
VK10.5%
 Microtiter plate:
Inject 10 μl test
solution seven
times.
2
Carryover
Alternately inject 10 μl
test solution and 10 μl
eluent three times.
PV 0.3%
3
Linearity
Inject 10, 20, 30, 40
and 50 μl test solution
three times each.
r 0.998
4
Mixture test
Inject 10 μl test
solution and 10 μl of
the dilution created by
the autosampler, three
times each.
VK2 0.5%
VK3 0.5%
F10: 9.85 < x < 10.25
Date:
Date of the next device test:
Tester:
Signature:
Result
56
Device test
Operation Qualification (OQ) with
ChromGate®
Extensive
functionality test
Start OQ
Extensive test of autosampler functionality using ChromGate®
chromatography software.
1. Select Control Qualification procedures. In the window 
that opens, select a performance qualification under 
Available Procedures.
2. In the Available procedures window, choose an OQ test for the
autosampler and choose Begin New Qualification Session.
3. Select [Start Session]. The Introduction window opens for the
function test.
4. Select [Continue] to display the required materials in the
chromatography software.
5. Select [Continue] to display the settings for the OQ test in 
the chromatography software.
Choose automatic
test
6. Select Perform all tests automatically.
7. Select Active channel of the detector.
8. Select Pump and click [Continue] to apply the settings.
Start OQ
Printing out the results
of the function test
Quit OQ
9. Click [Continue] to perform the function test. 
The chromatography software sets the flow rate to 
1 ml/min and the wavelength of the detector to 274 nm. 
You can print out the test results.
10. Select [Print report] to print the function test of the
autosampler.
11. Select [Exit] to end the function test of the device.
57
Device test
Operation Qualification Report
Legend
A Detailed information on autosampler
B Detailed function
test
A
C Date and fields for
signature and other
information
B
C
Fig. 50
Operation Qualification Report
58
Maintenance and care
Maintenance and care
KNAUER Technical Support
Contact data for
KNAUER Technical
Support
European hotline
E-mail:
If you have any technical questions regarding KNAUER hardware
or software, please use one of the contact options below:
KNAUER Technical Support Hotline
Languages: German and English
Available by telephone: 8 am to 5 pm (CET)
Phone:
+49-(0)30-809727-0
Fax:
+49-(0)30-8015010
[email protected]
Maintenance contract
The following maintenance work on the device may only be
performed by KNAUER or a company authorized by KNAUER
and is covered by a separate maintenance contract:
 Opening the device or removing housing parts
What maintenance tasks can users
perform on the device?
Users may perform the following maintenance tasks themselves:
 Exchanging the injection valve
 Exchanging the rotor seals
 Exchanging the syringe on the autosampler
 Exchanging the sample needle
 Exchanging the air needle
DANGER!
When covers are removed, potentially dangerous live components are exposed!
Switch off the device prior to maintenance tasks
and pull the power plug to completely isolate it
from the supply voltage!
The only time when the Smartline Autosampler
3950 need not be disconnected from the power
supply is if it is being checked exclusively by
means of the control software at the PC!
59
Maintenance and care
Leaks in the capillary screw fittings
Caution!
If leaks occur on the capillary screw fittings 
after maintenance and proper assembly, do not
tighten them further; instead replace them with
new connection capillaries.
Exchanging the fuses
1. Switch off the autosampler and remove the power plug to
completely disconnect the device from the power supply.
2. Remove the fuses from the fuse box at the rear of the device.
3. Insert new fuses (2x 2.5 A).
4. Plug in the power plug.
Exchanging the injection valve and
rotor seal
 Regularly clean the rotor seal of the injection valve.
 Regularly replace the rotor seal (approx. every three years).
 Remove the front panel of the autosampler.
 Remove the capillary connections, except the sample loop,
from the valve.
 During removal, consecutively loosen all screws by half a turn
respectively, until they can be removed.
Note:
Do not remove the screw in the cover plate hole
diagonal to the valve!
60
Maintenance and care
Removing the injection valve and rotor seal
Removing the injection
valve and rotor seal
Steps
1. With a screwdriver,
remove the screw
(A) on both sides of
the injection valve
support.
Figure
A
B
C
2. Remove the
injection valve.
3. With an Allen
wrench, remove
the screws (B) from
the stator (C).
Fig. 51
4. Carefully remove
the stator.
Removing the injection
valve
D
5. Remove the rotor
seal (D) from the
rotor (E).
6. Clean or exchange
the rotor seal.
Fig. 52
Removing the rotor seal
E
Fig. 53
Valve stator and rotor seal
61
Maintenance and care
Installing the rotor seal and injection valve
 During installation, hold the injection valve in such a way that
the bore hole for connecting the steel capillary to the pump
(port 1) is facing upward.
 Alternately tighten all screws by half a turn, until all screws
have been fully tightened.
Legend
A Labeling, port 1
A
B Connection from
capillary to pump
B
Fig. 54
Procedure
Installing the injection valve
1. Insert the rotor seal.
2. Place the valve stator block on the valve rotor and use an
Allen wrench to tighten the screws.
3. Insert the injection valve and use a screwdriver to tighten
the screw on both sides of the valve housing.
4. Connect the capillary again.
System flushing
Procedure
1. Connect the autosampler to the power supply.
2. Establish a connection to the PC.
3. Start the Smartline Autosampler 3950 Service Manager.
4. Select the Alias Direct Control menu.
5. Click Initialize to check whether the valve is correctly
positioned at the Inject position.
6. In the Initial wash field, click Start to flush the system.
7. In the Initial wash field, click Stop to stop flushing the system.
Exchanging the sample loop
By standard, the autosampler is equipped with a 100 μl sample
loop.
 When assembling a sample loop with a different injection
volume, make sure to use the correct combination of syringe
and capillaries and configure the controller software
appropriately.
 Always connect the sample loop to ports 2 and 5 of the
injection valve.
62
Maintenance and care
Exchanging the sample needle
 When using sample plates with 12 or 48 sample vials, 
make sure that the needle height setting is >2 mm to prevent
the needle from contacting the bottom of the sample vial.
 Only tighten the screw fitting until it is finger-tight, to
prevent the plastic capillary from becoming blocked.
Legend
A Screw fitting
A
B Plastic capillary
B
C Union nut
D Sample needle
C
D
Fig. 55
Procedure
Exchanging the sample needle
1. Start the Smartline Autosampler 3950 Service Manager.
2. Select the Alias Direct Control menu.
3. In the Needle field, click Exchange. The needle moves to the
replacement position.
4. Loosen the union nut (3).
5. Loosen the screw fitting (1) of the plastic capillary (2) on the
injection valve.
6. Remove the sample needle (4) with the plastic capillary.
7. Install a new sample needle unit. Make sure that the air seal
fully surrounds the sample needle.
8. Fasten the sample needle with the union nut.
9. Fasten the plastic capillary using the screw fitting on the
injection valve.
10. In the Direct Control window, click Initialize. The needle
moves to the initial position.
11. In the Initial wash field, click Start to flush the system.
12. In the Initial wash field, click Stop to stop flushing the
system.
13. Click Close to exit the Direct Control window.
63
Maintenance and care
14. Select the Alias Adjustments menu.
15. On the Needle-Tray tab, update the settings for the sample
plates.
Exchanging the air needle
 When exchanging the air needle, make sure that the thread
of the new height adjustment screw is flush with the lower
edge of the retaining nut.
 Make sure that the sealing ring is located in the retaining nut.
Legend
A Union nut
A
B Retaining nut
C Height adjustment
screw
D Air needle
B
E Sample needle
C
D
E
Fig. 56
Procedure
Exchanging the air needle
1. Start the Smartline Autosampler 3950 Service Manager.
2. Select the Alias Direct Control menu.
3. In the Needle field, click Exchange. The needle moves to the
replacement position.
4. Loosen the union nut (1).
5. Loosen the screw fitting of the plastic capillary on the
injection valve.
6. Remove the sample needle (5) with the plastic capillary.
7. Loosen the retaining nut (2) and pull it downwards together
with the air needle (4).
8. Unscrew the retaining nut from the height adjustment
screw (3).
9. Screw a new air needle with a new height adjustment screw
into the retaining nut.
10. Screw in the retaining nut.
11. Insert the sample needle and fasten with the union nut.
12. Fasten the plastic capillary using the screw fitting on the
injection valve.
64
Maintenance and care
13. In the Direct Control window, click Initialize. The needle
moves to the initial position.
14. In the Initial wash field, click Start to flush the system.
15. In the Initial wash field, click Stop to stop flushing the
system.
16. Click Close to exit the Direct Control window.
17. Select the Alias Adjustments menu.
18. On the Needle-Tray tab, update the settings for the sample
plates.
Exchanging the syringe
By standard, the autosampler is equipped with a 500 μl syringe.
 Use isopropanol as flushing solution to remove air bubbles
from the new syringe.
Legend
A Syringe valve
B Syringe
C Syringe drive
A
D Syringe plunger
B
C
D
Fig. 57
Procedure
Exchanging the syringe
1. Start the Smartline Autosampler 3950 Service Manager.
2. Select the Alias Direct Control menu.
3. In the Syringe field, click Exchange. The syringe plunger is
lowered.
4. Unscrew the syringe (2) by rotating it counterclockwise; 
leave the adapter in the syringe valve (1).
5. Remove the syringe plunger (4) from the syringe drive (3).
6. Fill new syringe with flushing solution.
7. Insert the syringe plunger into the syringe drive.
8. Tighten the syringe in the syringe valve by rotating it
clockwise.
9. In the Syringe field, click Home. The syringe content is
emptied into the drainage tube.
65
Maintenance and care
10. If there is still air in the syringe, click End in the Syringe field.
One syringe volume is aspirated into the syringe through the
flushing solution tube.
11. In the Syringe field, click Home. The syringe content is
emptied into the drainage tube.
12. Slightly tap the body of the syringe if it still contains air. 
If necessary, repeat step 10 and 11.
13. In the Initial wash field, click Start to flush the system.
14. In the Initial wash field, click Stop to stop flushing the
system.
15. Click Close to exit the Direct Control window.
Exchanging the syringe plunger or plunger tip
Procedure
1. Start the Smartline Autosampler 3950 Service Manager.
2. Select the Alias Direct Control menu.
3. In the Syringe field, click Exchange. The syringe plunger 
is lowered.
4. Remove the syringe (see above).
5. Pull the syringe plunger out of the glass cylinder of the
syringe.
6. Use a pair of tweezers to remove the plunger tip.
7. Wet the new plunger tip with isopropanol.
8. Mount the new plunger tip onto the syringe plunger.
9. Push the syringe plunger into the glass cylinder of the
syringe.
10. Install the syringe (see above.)
11. In the Syringe field, click Home. The syringe content is
emptied into the drainage tube.
66
Maintenance and care
Exchanging the syringe valve
The syringe valve has four connections, one of them remains
unused.
 Hand-tighten all fittings on the connections to the syringe
valve.
 To exchange the valve, set it to the Waste position, 
because the Allen screws are only accessible if the valve 
is in this position.
Legend
A Upper Allen screw
A
B
C
D
B Lower Allen screw
C Connector for flushing solution tube
(hidden)
E
F
D Connector for buffer tube
E Connector for
syringe
F Unused connection
Fig. 58
Procedure
Exchanging the syringe valve
1. Start Smartline Autosampler 3950 Service Manager
2. Select the Alias Direct Control menu.
3. In the Syringe field, click Exchange. The syringe plunger 
is lowered.
4. Loosen with two turns the lower Allen screw.
5. Loosen with two turns the upper Allen screw.
6. Pull out the upper part of the syringe.
7. Remove the syringe.
8. Exchange the syringe valve.
9. Insert a new syringe.
10. Tighten the Allen screws.
67
Maintenance and care
Cleaning and caring for the device
DANGER!
Risk of electrical shock or short circuit if cleaning
solution enters the device's interior! Only lighty
moisten the cleaning cloth!
All smooth surfaces of the device can be cleaned with a mild,
commercially available cleaning solution, or with isopropanol.
 Use a soft cloth to clean the collecting container and to
remove any spilled sample solution from the sample plate
 To remove deposits, clean the drainage tube regularly using
solvent
68
Environmental protection
Environmental protection
Disposal
Drop the devices off at the local municipal waste facilities or
send the devices back to the manufacturer where it will be disposed of properly.
Decontamination
Contamination of devices with toxic, infectious or radio-active
substances poses a hazard for all persons during operation,
repair, sale and disposal of a device.
DANGER!
Danger caused by toxic, infectious, or radioactive substances! A contaminated device must
never be submitted for repairs, sold, or disposed
of!
Contract a specialist company to decontaminate
the device or perform the decontamination
yourself if you have the required expertise!
All contaminated devices must be properly decontaminated by
a specialist company or the operating company before they can
be recommissioned, repaired, sold, or disposed of.
All materials or fluids used for decontamination must be collected separately and disposed of properly.
69
Troubleshooting
Troubleshooting
Device errors
One possible cause of device errors is a malfunctioning valve.
Checking the valve
Remove the valve and check all parts for wear and
contamination. After the problem has been eliminated and 
the valve reinstalled, perform the following steps:
Procedure
1. Select the Alias Direct Control menu.
2. In the Direct Control window, click Initialize. The needle
moves to the initial position.
3. In the Initial wash field, click Start to flush the system.
4. In the Initial wash field, click Stop to stop flushing the system.
5. Click Close to exit the Direct Control window.
Software fault
Software errors can occur due to faulty communication between
the devices or incorrect installation of the software.
Procedure
1. Check the cable connections.
2. Start PLATINblue Autosampler AS-1 Service Manager.
3. Select Alias Direct Control.
4. In the Direct Control window, click Initialize.
Analytical errors
Possible causes:
 Wear due to errors in the injection and method settings.
 Unsuitable combination of sample loop, buffer tube and
syringe.
 External effects such as temperature, and light-sensitive
samples being exposed to light.
Solutions:
 Check whether the application has run previously without
errors and that no changes have been made to the analytical
system.
 Determine whether the fault is caused by the autosampler or
other devices in the system.
70
Troubleshooting
If the required degree of reproducibility is not achieved, 
check the following possible sources of error and instigate 
steps to eliminate them:
Cause of fault
Elimination
Air in liquid path
Initialize the Smartline Autosampler
3950.
Leaking syringe
 If the syringe is leaking at the top,
check whether it has been installed
correctly.
 If the syringe is leaking at the
bottom, exchange the syringe
plunger tip.
Leaking syringe
valve
Check valve and exchange if required.
Rotor seal worn
Exchange the rotor seal and check the
stator block of the valve.
Dead volume in
capillary
connections
Install new fittings onto capillary
connections.
If an empty sample run returns an excessively large peak, 
check the following possible causes of error and instigate 
steps to eliminate them:
Cause of fault
Elimination
Solubility problems
Either modify sample or accept
carryover.
Interaction between
the empty sample
and the hardware
 Check hardware:
 Flush needle (inside and
outside) or install a different
needle type (steel, PEEK or with
glass coating).
 Injection valve: Exchange rotor
seal (other material).
 Capillaries and tubing: Use
other connections between the
autosampler and the columns
(steel, PEEK) or other flushing
solutions.
Empty sample
contaminated
Use new empty sample.
Cause unknown.
Attempt to solve problem by using
different solvents and eluents.
71
Troubleshooting
If no injection is performed:
Cause of fault
Elimination
Liquid path
blocked
1. Disconnect the plastic capillary of the
needle from the injection valve.
2. Start system flushing.
3. If solvent escapes at the injection valve
connection to the needle, check the
needle.
4. If no solvent escapes at the injection
valve connection to the needle,
disconnect the buffer tube from the
injection valve.
5. Start system flushing.
6. If solvent flows out at the open end of
the buffer tube, check the rotor seal.
7. If no solvent flows out of the open end
of the buffer tube, disconnect the buffer
tube from the syringe valve.
8. Start system flushing.
9. If solvent flows out of syringe valve,
check the buffer tube.
10. If no solvent flows out of the syringe
valve, check whether the connections of
the liquid path have been tightened too
much.
Leaking
injection valve
1. Unscrew the plastic capillary leading to
the needle from the injection valve.
2. Disconnect the plastic capillary leading
to the syringe from the injection valve.
C Connect the HPLC pump to the injection valve.
D Close the connection to the column at
the injection valve.
E Start the pump at a low flow rate.
F Check the connections to the syringe
and to the needle at the injection valve
to ensure that they are tight.
G If liquid escapes there, check the rotor
seal.
H If no liquid escapes there, check the
HPLC with a manual valve.
72
Troubleshooting
System messages from ChromGate®
The various system messages of the ChromGate®
chromatography software from KNAUER are explained below.
The system messages are sorted alphabetically.
System message
Explanation
Autosampler is in run mode.
 Quit the controller software
and restart.
 Switch the devices off and on
again.
Autosampler is not
responding. Please check
communication settings
and ensure the device is
online.
 Switch the devices off and on
again.
Cannot run autosampler.
 Switch the devices off and on
again.
 Check the network settings. If
the system message appears
again, notify KNAUER
Technical Support.
 Check the network settings. If
the system message appears
again, notify KNAUER
Technical Support.
Cannot set destination vial
to %d.
Check parameters in controller
software and correct entry.
Cannot set first transport
vial to %d.
Check parameters in controller
software and correct entry.
Cannot set last transport
vial to %d.
Check parameters in controller
software and correct entry.
Cannot stop autosampler.
Check the network settings.
If the system message appears
again, notify KNAUER Technical
Support.
Communication port for
autosampler was not
initialized. Please check the
configuration settings.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Configuration settings do
not match with the device.
Run cannot start.
Check configuration and
settings.
Destination position not
reached.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
73
Troubleshooting
System message
Explanation
Deviation of more than
+/-2 mm towards home.
 Look for visible obstructions
in area of sample plate.
 Check the belt tension of the
tray.
Dispenser error.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
EEPROM error in
adjustments.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
EEPROM error in log
counter.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
EEPROM error in settings.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
EEPROM write error.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Electronics error.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error 369
Not enough transport liquid. 
Fill up transport liquid.
Error 370
Not enough reagent. 
Fill up reagent.
Error by setting Mix&Dilute
vials.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error occurred during
initialization, the
autosampler cannot start.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error resetting output.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
74
Troubleshooting
System message
Explanation
Error running user defines
program.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting injection
mode.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting injection
mode.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting needle height.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting syringe speed.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting the analysis
time.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting the auxiliaries.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting the flush time.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting the flush
volume.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting the injection
volume.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting the loop
volume.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
75
Troubleshooting
System message
Explanation
Error setting the prep.
mode.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting the syringe
volume.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting the tray
configuration.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting the tray
temperature.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting the vial
number.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting timed events.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting tubing volume.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Error setting wash volume.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Flush volume error.
Check parameters in controller
software and correct entry.
Home sensor activated
when not expected.
 Check parameters in
controller software and
correct entry.
 Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
76
Troubleshooting
System message
Explanation
Home sensor not 
de-activated.
 Check whether there are
visible obstructions impairing
the sample plate tray.
 Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Home sensor not reached.
 Check whether there are
visible obstructions impairing
the sample plate tray.
 Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Horizontal: Home sensor
activated when not
expected.
 Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Horizontal: Home sensor
not de-activated.
 Check whether there are
visible obstructions impairing
the needle unit.
 Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Horizontal: Home sensor
not reached.
 Check whether there are
visible obstructions impairing
the needle unit.
 Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Horizontal: Needle position
is unknown.
Initialize the needle unit using
the controller software.
Illegal sensor readout.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Incorrect first destination
vial.
Check parameters in controller
software and correct entry.
77
Troubleshooting
System message
Explanation
Injection needle unit error.
 Check whether there are
visible obstructions impairing
the needle unit.
 Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Injection valve or ISS unit
error.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Injection volume %.2f is
invalid. For specified
injection method, volume
should be within the range
%.2f μl-%.2f μl, with
%2f. μl increments.
Check parameters in controller
software and correct entry.
Injection volume error.
Check parameters in controller
software and correct entry.
Invalid %s vial position
%02d. The vial position
must be between 01 and
%02d.\n.
Check parameters in controller
software and correct entry.
Invalid combination of the
trays. The combination of
different trays for the
Mix&Dilute mode is not
allowed.
 Insert the correct sample
plate.
Invalid combination of the
trays. The combination of
plates 384 low and 96 high
is not allowed.
 Insert the correct sample
plate.
Invalid configuration. 
ISS option not installed on
autosampler. Please switch
off this option in
configuration dialog.
Check parameters in controller
software and correct entry.
Invalid configuration. 
SSV option not installed on
autosampler. Please switch
off this option in
configuration dialog.
Check parameters in controller
software and correct entry.
 Check parameters in
controller software and
correct entry.
 Check parameters in
controller software and
correct entry.
78
Troubleshooting
System message
Explanation
Invalid flush volume %2f μl.
The flush volume should be
between 0 and %2f μl.
Check parameters in controller
software and correct entry.
Invalid flush volume %d μl.
The flush volume should be
between 0 and %d μl.
Check parameters in controller
software and correct entry.
Invalid input. Only values
with increments of %.2f
allowed.
Check parameters in controller
software and correct entry.
Invalid instrument is
detected.
Check parameters in controller
software and correct entry.
Invalid integer number.
Check parameters in controller
software and correct entry.
Invalid loop volume %2f μl.
The loop volume should be
between 0 and %2f μl.
Check parameters in controller
software and correct entry.
Invalid loop volume %d μl.
The loop volume should be
between 0 and %d μl.
Check parameters in controller
software and correct entry.
Invalid mix program: No
Destination vial is specified
in configuration dialog.
Check parameters in controller
software and correct entry.
Invalid mix program: No
reagent A vial is specified in
configuration dialog.
Check parameters in controller
software and correct entry.
Invalid mix program: No
reagent B vial is specified in
configuration dialog.
Check parameters in controller
software and correct entry.
Invalid mix times. The time
should be between 1 and 9.
Check parameters in controller
software and correct entry.
Invalid needle height
%d mm. The needle height
should be between %d and
%d mm.
Check parameters in controller
software and correct entry.
Invalid time-based method.
Several AUX events have
the same time.
Check parameters in controller
software and correct entry.
Invalid time-based method.
Several SSV events have the
same time.
Check parameters in controller
software and correct entry.
Invalid tray configuration:
two or more vial positions
are the same.
Check parameters in controller
software and correct entry.
79
Troubleshooting
System message
Explanation
Invalid tray temperature
%d ˚C. The temperature
should be between 4 and
22 ˚C.
Check parameters in controller
software and correct entry.
Invalid tubing volume
%2f μl. The tubing volume
should be between %2f and
%2f μl.
Check parameters in controller
software and correct entry.
Invalid tubing volume
%d μl. The tubing volume
should be between 0 and
%d μl.
Check parameters in controller
software and correct entry.
Invalid volume %d μl. 
The volume should be
between the 0 and the
syringe volume (%d μl).
Check parameters in controller
software and correct entry.
Invalid wait time. The time
should be between 0 and
9 h 50 min 59 sec. Invalid
wash volume %d μl. The
wash volume should be
between %d and %d μl
Check parameters in controller
software and correct entry.
ISS valve error.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
ISS-A option not installed
on autosampler. Please
switch off ISS-A option in
configuration dialog.
Check controller software
configuration and correct entry.
ISS-B option not installed
on autosampler. Please
switch off ISS-B option in
configuration dialog.
Check controller software
configuration and correct entry.
Missing destination vial.
 Check position of sample vial.
 Check parameters in
controller software and
correct entry.
Missing reagent vial.
 Check position of sample vial.
 Check parameters in
controller software and
correct entry.
80
Troubleshooting
System message
Explanation
Missing transport vial.
 Check position of sample vial.
 Check parameters in
controller software and
correct entry.
Missing vial.
 Check position of needle unit.
 Switch the devices off and on
again.
Missing wash vial error.
 Check position of needle unit.
 Switch the devices off and on
again.
Needle movement error.
 Check position of needle unit.
 Switch the devices off and on
again.
No destination vial is
specified in the
configuration.
Check parameters in controller
software and correct entry.
No reagent A vial is
specified in the
configuration.
Check parameters in controller
software and correct entry.
No reagent B vial is
specified in the
configuration.
Check parameters in controller
software and correct entry.
No transport vials are
defined in the tray
configuration. It is not
possible to use the μl pickup injection mode.
Check parameters in controller
software and correct entry.
No user defined or mix
program is running.
Check parameters in controller
software and correct entry.
Not enough reagent liquid.
Check volume of liquid and
change as required.
Not enough transport liquid
available due to missing
transport vials.
Check volume of liquid and
change as required.
Oven option not installed
on autosampler. Please
switch off oven option in
configuration dialog.
Check controller software
configuration and correct entry.
Please specify inject marker
or AUX event to be able to
trigger the run.
Check parameters in controller
software and correct entry.
81
Troubleshooting
System message
Explanation
Selecting transport position
failed.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Serial number is not valid.
Please check the
configuration.
Check parameters in controller
software and correct entry.
Setting mix program error.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Setting service mode failed.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Syringe dispenser unit error.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Syringe home sensor not
de-activated.
 Needle flushing with
controller software.
 Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Syringe home sensor not
reached.
 Needle flushing with
controller software.
 Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Syringe position is
unknown.
Initialize the syringe unit using
the controller software.
Syringe rotation error.
 Needle flushing with
controller software.
 Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Syringe valve did not find
destination position.
 Needle flushing with
controller software.
 Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
82
Troubleshooting
System message
Explanation
Temperature above 48 ˚C
at cooling ON.
 Switch off the cooling and
check whether ambient
temperature sensor is
properly functioning.
 If the system message
appears again, notify KNAUER
Technical Support.
The 10 ml syringe cannot
be used for standard
injections.
Exchange the syringe.
The autosampler has
detected another tray than
that which is currently
configured. Please select the
correct tray in configuration
dialog.
Check controller software
configuration and correct entry.
The autosampler is not
ready. Please try later.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
The injection volume of
%2f μl is invalid. For the
specified injection method,
volume should equal
%2f μl.
Check parameters in controller
software and correct entry.
The sample needle is not in
the home position while the
tray is rotating.
Check parameters in controller
software and correct entry.
Trace from tray cooling
cannot be acquired. Tray
cooling is off.
Check parameters in controller
software and correct entry.
Tray advance is not
available at this time.
Check parameters in controller
software and correct entry.
Tray error.
Check parameters in controller
software and correct entry.
Valve error.
Check parameters in controller
software and correct entry.
Vertical: Home sensor not
de-activated.
 Check whether there are
visible obstructions impairing
the needle unit.
 Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
83
Troubleshooting
System message
Explanation
Vertical: Home sensor not
reached.
 Check whether there are
visible obstructions impairing
the needle unit.
 Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Vertical: Needle position is
unknown.
Initialize the instrument in the
controller software.
Vertical: stripper did not
detect plate (or wash/
waste). Missing vial.
 Check sample vial and plate.
Vertical: stripper stuck.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Vertical: The sample needle
arm is at an invalid
position.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
Vial number error.
Check parameters in controller
software and correct entry.
Wear-out limit reached.
Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support. The injection
valve must be replaced.
Wrong loop volume. 
The largest loop volume for
standard injections is
1000 μl.
Check parameters in controller
software and correct entry.
Wrong tubing volume. 
The largest tubing volume
for standard injections is
200 μl.
Check parameters in controller
software and correct entry.
 Switch the devices off and on
again. If the system message
appears again, notify KNAUER
Technical Support.
84
Technical data
Technical data
Ambient
conditions
Temperature range
10–40 ˚C; 39–104 ˚F
Air humidity
20–80% air humidity
(non-condensing)
Smartline
Autosampler 3950
Standard version
Sample capacity
Microtiter plates for max.
768 samples or sample plates 
for max. 96 vials
Injection volume
1–5000 μl
Standard sample loop
100 μl
Syringe
500 μl
Injection time
<60 s, incl. cleaning
Injection modes
PASA™ loop injection principle:
 Full loop injection
 Partial loop injection
 Microliter pickup injection
Reproducibility
RSD (Relative Standard Deviation):
 full loop injection: <0.3%
 partial loop injection at an
injection volume >10 μl: <0.5%
 microliter pickup injection at an
injection volume >10 μl: <1.0%
Carryover
<0.05% with needle cleaning
Sample cooling
4–22 °C
Weight
 19 kg
 With cooling: 21 kg
Dimensions
(length x width x
height)
 510 mm x 300 mm x 360 mm
Supply voltage range
95–240 V
Supply frequency
50–60 Hz
 With cooling:
575 mm x 300 mm x 360 mm
85
Technical data
Preparative version
Sample capacity
2x sample plate with 12 vials
respectively (10 ml)
Vial height
32–47 mm
Injection volume
1–10000 μl
Standard sample loop
10 ml
Syringe
2500 μl
Injection time
<60 s, incl. cleaning
Injection modes
PASA™ loop injection principle:
Partial loop injection
Reproducibility
RSD:
partial loop injection at an injection
volume >10 μl: <1%
Carryover
<0.1% with needle cleaning
Weight
19 kg
Dimensions
(length x width x
height)
510 mm x 300 mm x 360 mm
Supply voltage range
100–240 V
Supply frequency
50–60 Hz
86
Delivery program
Delivery program
Devices and accessories
Name
Order number
Smartline Autosampler 3950,
standard version, injection valve,
accessories
A5005-1
Smartline Autosampler 3950,
standard version incl. sample cooling,
accessories
A50051-1
Smartline Autosampler 3950,
biocompatible version, accessories
A50052-1
Smartline Autosampler 3950,
biocompatible version incl. sample
cooling, accessories
A50053-1
Smartline Autosampler 3950,
preparative version, incl. vial plates for
12 x 10 vials, accessories
A50054-1
Manual
V1509A
Spare parts
Name
Order number
Autosampler accessory kit:
A0664
 Sample vial with closure and septa
(1.5 ml, 1000 pcs.)
 Opening and closing pliers
2x sample plate for 48 vials, 1.5 ml
A50050
Vial plate for 12 vials,
10 ml (preparative version)
M2072
Microtiter plate 96 wells,
U sanitized, 0.35 ml
A1823
Microtiter plate 96 wells,
U sanitized, 1.2 ml
A1823V1
PTFE tube, 3.2 mm AD
1.5 mm ID, 300 cm
A0732
Silicon tube
8.0 mm ID, 200 cm
M20702
Flushing solution bottle
M2054
87
Delivery program
Name
Order number
Syringe 500 μl
M2070
Syringe 2500 μl
M20701
Sample needle kit for SPARK valve 1/16"
A64700
Sample needle
A0646
Sample needle incl. tube and connector,
biocompatible version
A15086
Air needle, white, 62 mm
A50058
Set of air needles
A50059
Air needle, yellow, 50 mm
M20401
Air needle, red, 56 mm
M20402
Air needle, blue, 68 mm
M20403
Air needle, green, 74 mm
M20404
Air needle, black, 80 mm
M20405
2x 2.5 A fuse
M2040
Network cable
A5255
RS-232 cable
A0895
Manual
V1509A
Accessories 84+3
Name
Order number
Vial tray for 84x1.5ml and 3x10ml vials
A500501
125 Vials 10 ml, 500 crimp caps and 500
septa, ø 22 mm
A1662
88
Legal information
Legal information
Warranty conditions
The factory warranty for the device is valid for 12 months after
the date of dispatch. All warranty claims shall expire in the event
that any unauthorized changes are made to the device.
During the warranty period, any components with material or
design-related defects will be replaced or repaired by KNAUER
free of charge.
This warranty excludes the following:
1. Accidental or willful damage
2. Damage or errors caused by third parties that are not
contractually related to KNAUER at the time the damage
occurs
3. Wear parts, fuses, glass parts, columns, light sources, cuvettes
and other optical components
4. Damage caused by negligence or improper operation of the
devices and damage caused by clogged capillaries
5. Packaging and transport damage
In the event of device malfunctions, contact:
Manufacturer
Wissenschaftliche Gerätebau
Dr. Ing. Herbert KNAUER GmbH
Hegauer Weg 38
14163 Berlin, Germany
Phone:
+49–(0)30–809727–0
Fax:
+49–(0)30–8015010
E-Mail:
[email protected]
Internet: www.knauer.net
The packaging of our devices provides the best possible
protection against transport damage. However, immediately
inspect each delivery for signs of transport damage. If the
shipment is incomplete or damaged, inform the manufacturing
factory within three workdays. Also inform the freight carrier
about transport damage.
89
Legal information
Declaration of conformity
Manufacturer name
and address
Smartline
Autosampler 3950
Wissenschaftliche Gerätebau
Dr. Ing. Herbert KNAUER GmbH
Hegauer Weg 38
14163 Berlin, Germany
Order number A5005-1, A50051-1, A50052-1,A50053-1,
A50054-1
complies with the following requirements and product
specifications:
 DIN EN 60799 (June 1999) Electrical accessories – Cord sets
and interconnection cord sets
 DIN EN 61010-1 (August 2002) Safety requirements for
electrical equipment for measurement, control and
laboratory use
 Low voltage directive (2006/95/EC)
 DIN EN 61000-3-2 (March 2010) Electromagnetic
compatibility (EMC) Part 3-2
EN 61000-3-2:2006 + A1:2009 + A2:2009
 EMC standarts (2004/108/EC)
 DIN EN 61326-1 (October 2006) Electrical equipment for
measurement, control and laboratory use – EMC
requirements
 DIN EN 61326-1 Corrigendum 2 (April 2011)
 Directives for an environmentally sound use of electrical and
electronic equipment
 RoHS directive 2002/95/EC (February 2003) on the
restriction of the use of certain hazardous substances in
electrical and electronic equipment
 WEEE directive 2002/96/EC (February 2003) on waste
electrical and electronic equipment
Date
Berlin, 2012-01-04
Dr. Alexander Bünz (Managing Director)
The mark of conformity has been applied to the rear panel of
the device.
90
Abbreviations and terminology
Abbreviations and terminology
Here you can find information on the abbreviations and
terminology used in this device manual for the Smartline
Autosampler 3950.
Terminology
Explanations
GLP
Good Laboratory Practice – quality assurance
for laboratories.
HPG
High Pressure Gradient (HPG). Operating
mode of an HPLC or UHPLC system.The solvent is mixed on the high-pressure side of the
pump.
HPLC
High Pressure Liquid Chromatography
(HPLC).
IP address
Unique address of transmitter or receiver in
local network or Internet (Internet protocol).
Solvent
Mobile phase (eluent) or carrier for liquid
chromatography.
LPG
Low-pressure gradient, LPG.Operating mode
of an HPLC or UHPLC system. The solvent is
mixed on the low-pressure side of the pump.
Remote
The chromatography software controls the
autosampler.
UHPLC
Ultra High-Pressure Liquid Chromatography
(UHPLC). Ultra high-pressure liquid
chromatography.
91
Table of Figures
Fig. 1:
Fig. 2:
Fig. 3:
Fig. 4:
Fig. 5:
Fig. 6:
Fig. 7:
Fig. 8:
Fig. 9:
Fig. 10:
Fig. 11:
Fig. 12:
Fig. 13:
Fig. 14:
Fig. 15:
Fig. 16:
Fig. 17:
Fig. 18:
Fig. 19:
Fig. 20:
Fig. 21:
Fig. 22:
Fig. 23:
Fig. 24:
Fig. 25:
Fig. 26:
Fig. 27:
Fig. 28:
Fig. 29:
Fig. 30:
Fig. 31:
Fig. 32:
Fig. 33:
Fig. 34:
Fig. 35:
Fig. 36:
Fig. 37:
Fig. 38:
Push the door into the interior . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Removing the front panel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Removing the cooler cover . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Front view of device with sample compartment . . . . . . . . . . . . . . .
Rear view of the autosampler . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
PASA™ loop injection principle . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Full loop: Initial situation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Full loop: The needle and sample lines are flushed . . . . . . . . . . . . .
Full loop: Injection valve switches to LOAD position . . . . . . . . . . . . .
Full loop: Sample loop is filled completely . . . . . . . . . . . . . . . . . . . .
Full loop: Injection valve switches to INJECT position . . . . . . . . . . . .
Full loop injection with air segment (A), 
without air segment (B) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Partial loop: Initial situation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Partial loop: The sample line is filled with sample . . . . . . . . . . . . . .
Partial loop: Injection valve switches to LOAD position . . . . . . . . . .
Partial loop: Sample loop is filled partially . . . . . . . . . . . . . . . . . . . .
Partial loop: Injection valve switches to INJECT position . . . . . . . . . .
Partial loop injection with air segment (A), 
without air segment (B) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Microliter pick-up: Washing liquid is aspirated . . . . . . . . . . . . . . . .
Microliter pick-up: Injection valve switches to LOAD position . . . . . .
Microliter pick-up: The needle moves to sample vial/well . . . . . . . .
Microliter pick-up: Injection volume of sample is aspirated . . . . . . .
Microliter pick-up: Sample is transported into sample loop . . . . . . .
Microliter pick-up: Sample is transported to column . . . . . . . . . . . .
Microliter pick-up injection with air segment (A), 
without air segment (B) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Microliter pick-up 84+3: Initial situation . . . . . . . . . . . . . . . . . . . . .
Microliter pick-up 84+3: Sample line is filled with transport liquid . .
Microliter pick-up 84+3: The sample is aspirated . . . . . . . . . . . . . . .
Microliter pick-up 84+3: Sample is transported into sample loop . .
Microliter pick-up 84+3: Injection valve switches to INJECT position.
Height 84+3 tray . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Needle height and fluid levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Processing order . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Transport/Reagent positions on 84+3 tray . . . . . . . . . . . . . . . . . . . .
Standard air needle with 10 and 1.5 ml sample vial . . . . . . . . . . . .
Standard air needle with Greiner microtiter plates . . . . . . . . . . . . . .
Calculating the correct air needle . . . . . . . . . . . . . . . . . . . . . . . . . .
Selected air needles with sample vials . . . . . . . . . . . . . . . . . . . . . . .
16
16
16
17
17
18
20
20
20
21
21
21
22
22
22
23
23
24
25
25
26
26
26
26
27
28
28
28
29
29
30
30
31
31
33
33
34
35
92
Fig. 39:
Fig. 40:
Fig. 41:
Fig. 42:
Fig. 43:
Fig. 44:
Fig. 45:
Fig. 46:
Fig. 47:
Fig. 48:
Fig. 49:
Fig. 50:
Fig. 51:
Fig. 52:
Fig. 53:
Fig. 54:
Fig. 55:
Fig. 56:
Fig. 57:
Fig. 58:
Selected air needles with Greiner microtiter plates . . . . . . . . . . . . .
Sample positions in columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Sample positions in rows . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Transport/Reagent positions on 84+3 tray . . . . . . . . . . . . . . . . . . . .
Connection schematic for capillaries and tubing . . . . . . . . . . . . . . .
Connections on the injection valve . . . . . . . . . . . . . . . . . . . . . . . . .
Syringe connections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Tube guide for flushing solution tube . . . . . . . . . . . . . . . . . . . . . . .
Connecting the drainage tubing . . . . . . . . . . . . . . . . . . . . . . . . . . .
Autosampler configuration with ClarityChrom® . . . . . . . . . . . . . . .
System flushing with the Service Manager software . . . . . . . . . . . .
Operation Qualification Report . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Removing the injection valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Removing the rotor seal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Valve stator and rotor seal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Installing the injection valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Exchanging the sample needle . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Exchanging the air needle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Exchanging the syringe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Exchanging the syringe valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
35
38
39
40
42
42
43
43
44
45
47
57
60
60
60
61
62
63
64
66
93
Index
A
abbreviations 90
air needle 33
air needle types 34
standard air needle 33
C
Care 67
Cleaning 67
connection
drainage tubing 43
I/O connection 47
closed-contact output 48
TTL inputs 48
injection valve 42
LAN 7, 45
syringe 43
D
Dead volume 10
declaration of conformity 89
Decontamination 68
device test 49
archiving 54
linearity 52
mixture test 53
operation qualification with ChromGate® 56
reproducibility 50
sample carryover 51
test interval 49
Disposal 68
E
Environmental protection 68
F
filter 11
Flammability 9
flashpoint 9
G
gradient grade, filtered solvents 11
I
installation 13
K
KNAUER technical service 58
L
laboratory use 7
Leaks
on capillary screw fittings 59
M
mains connection 10
maintenance 58
exchange syringe valve 66
exchanging air needle 63
exchanging fuses 59
exchanging injection valve and rotor seal 59
exchanging sample loop 61
exchanging sample needle 62
exchanging syringe 64
exchanging syringe plunger or plunger tip 65
maintenance contract 58
mixing and thinning
Add 37
Mix 37
sample positions 38
O
operation
System 10
P
PASA™ loop injection principle 18
microliter pick-up injection 25
PEEK 9
power supply 10
protective measures 10
S
safety 8
laboratory regulations 8
scope of supply 13
self-ignition point 9
software 45
ClarityChrom® 45
Smartline Autosampler 3950 Service Manager
46
solvents 8
flushing solution 9
solvent tray 9
toxicity 9
unsuitable 9
Solvents, suitable 8
symbols 12
system flushing 46
Smartline Autosampler 3950 Service Manager
94
46
T
target group 10
technical data 84
toxicity
solvents 9
Troubleshooting 69
troubleshooting
analytical errors 69
ChromGate® system messages 72
software errors 69
U
UHPLC system
operation 10
unsuitable solvents 9
W
warranty conditions 88
© Wissenschaftliche Gerätebau
Dr. Ing. Herbert Knauer GmbH
All rights reserved.
The information in this document is subject to change without prior notice.
Translation of the original German edition of this manual.
2012-01-04
Printed in Germany.
® ChromGate and ClarityChrom
are registered trademarks of
Wissenschaftliche Gerätebau
Dr. Ing. Herbert Knauer GmbH
www.knauer.net
HPLC · SMB · Osmometry
Wissenschaftliche Gerätebau
Dr. Ing. Herbert Knauer GmbH
Hegauer Weg 38
D-14163 Berlin, Germany
Phone:+49-(0)30-809727-0
Telefax:+49-(0)30-8015010
E-Mail:[email protected]
Internet:www.knauer.net
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