ÄKTA
User Manual
™
pure
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Table of Contents
Table of Contents
71 Introduction ......................................................................................................................................................
81.1 Important user information ............................................................................................................................................
101.2 ÄKTA pure overview ...........................................................................................................................................................
121.3 ÄKTA pure user documentation ....................................................................................................................................
142 The ÄKTA pure instrument .............................................................................................................................
152.1 Overview illustrations ........................................................................................................................................................
262.2 Liquid flow path ...................................................................................................................................................................
282.3 Instrument control panel .................................................................................................................................................
332.4 Instrument modules ...........................................................................................................................................................
342.4.1 System pumps ......................................................................................................................................................................................................
382.4.2 Mixer .........................................................................................................................................................................................................................
392.4.3 Valves, overview ..................................................................................................................................................................................................
402.4.4 Inlet valves .............................................................................................................................................................................................................
472.4.5 Mixer valve .............................................................................................................................................................................................................
502.4.6 Injection valve .......................................................................................................................................................................................................
532.4.7 Loop valve ..............................................................................................................................................................................................................
562.4.8 Column valves ......................................................................................................................................................................................................
602.4.9 Versatile valve .......................................................................................................................................................................................................
612.4.10 pH valve ...................................................................................................................................................................................................................
652.4.11 Outlet valves ..........................................................................................................................................................................................................
682.4.12 Pressure monitors ...............................................................................................................................................................................................
702.4.13 UV monitors ...........................................................................................................................................................................................................
732.4.14 Conductivity monitor .........................................................................................................................................................................................
752.4.15 Flow restrictor .......................................................................................................................................................................................................
772.5 Installation of internal modules ....................................................................................................................................
802.6 Accessories ............................................................................................................................................................................
873 ÄKTA pure external modules ..........................................................................................................................
883.1 External air sensors ............................................................................................................................................................
903.2 Fraction collector F9-C .....................................................................................................................................................
913.2.1 Function ..................................................................................................................................................................................................................
933.2.2 Fraction collector F9-C illustrations ............................................................................................................................................................
973.2.3 Cassettes, Cassette tray and racks .............................................................................................................................................................
1033.2.4 Connect tubing to the ÄKTA pure instrument .........................................................................................................................................
1043.3 Fraction collector F9-R ....................................................................................................................................................
1083.4 Sample pump S9 and S9H ..............................................................................................................................................
1123.5 I/O-box E9 ...............................................................................................................................................................................
1133.5.1 Overview of the I/O-box ...................................................................................................................................................................................
1153.5.2 Analog connector and signals .......................................................................................................................................................................
1173.5.3 Digital connector and signals ........................................................................................................................................................................
1193.5.4 Connect external equipment to the I/O-box ...........................................................................................................................................
1223.6 Connection of external modules ..................................................................................................................................
1244 System configuration ......................................................................................................................................
1254.1 Configuration overview ....................................................................................................................................................
1314.2 Configure modules .............................................................................................................................................................
1324.2.1 Configuration of inlet valves ..........................................................................................................................................................................
1334.2.2 Configuration of Mixer valves ........................................................................................................................................................................
1344.2.3 Configuration of Loop valves .........................................................................................................................................................................
1354.2.4 Configuration of column valves ....................................................................................................................................................................
ÄKTA pure User Manual 29119969 AB 3
Table of Contents
1374.2.5 Configuration of Versatile valves ..................................................................................................................................................................
1384.2.6 Configuration of pH valves .............................................................................................................................................................................
1394.2.7 Configuration of outlet valves .......................................................................................................................................................................
1404.2.8 Configuration of UV monitors ........................................................................................................................................................................
1414.2.9 Configuration of Conductivity monitor ......................................................................................................................................................
1424.2.10 Configuration of external air sensors .........................................................................................................................................................
1434.2.11 Configuration of fraction collectors ............................................................................................................................................................
1454.2.12 Configuration of I/O-box ..................................................................................................................................................................................
1464.3 General system settings ..................................................................................................................................................
1495 Operation ..........................................................................................................................................................
1505.1 Before you prepare the system ....................................................................................................................................
1515.2 Prepare the flow path ........................................................................................................................................................
1585.3 Start UNICORN and connect to system ....................................................................................................................
1615.4 Prime inlets and purge pump heads ..........................................................................................................................
1625.4.1 System pumps ......................................................................................................................................................................................................
1685.4.2 Sample pump .......................................................................................................................................................................................................
1735.5 Connect a column ...............................................................................................................................................................
1775.6 Pressure alarms ...................................................................................................................................................................
1805.7 Sample application .............................................................................................................................................................
1815.7.1 Sample application using direct injection onto the column .............................................................................................................
1835.7.2 Sample application using a Superloop™...................................................................................................................................................
1905.7.3 Sample application using a sample loop .................................................................................................................................................
1935.8 Fractionation .........................................................................................................................................................................
1945.8.1 Prepare Fraction collector F9-C ....................................................................................................................................................................
2005.8.2 Prepare Fraction collector F9-R ....................................................................................................................................................................
2075.8.3 Fractionation overview .....................................................................................................................................................................................
2115.9 Create a method and perform a run .........................................................................................................................
2125.9.1 Create a method ..................................................................................................................................................................................................
2175.9.2 Prepare and perform a run .............................................................................................................................................................................
2195.9.3 Monitor a run ........................................................................................................................................................................................................
2215.9.4 After run procedures .........................................................................................................................................................................................
2256 Performance tests ...........................................................................................................................................
2266.1 General performance test actions ..............................................................................................................................
2296.2 Air sensor A and Inlet valve A tests .............................................................................................................................
2316.3 Air sensor B and Inlet valve B tests .............................................................................................................................
2336.4 Air sensor S and Sample inlet valve tests ................................................................................................................
2356.5 Column valve C tests .........................................................................................................................................................
2376.6 Fraction collector F9-C test ............................................................................................................................................
2406.7 Fraction Collector F9-R Test ...........................................................................................................................................
2426.8 Sample pump tests ............................................................................................................................................................
2446.9 System Test UV U9-L (fixed) ............................................................................................................................................
2486.10 System Test UV U9-M (variable) ....................................................................................................................................
2527 Maintenance .....................................................................................................................................................
2537.1 Maintenance Manager .....................................................................................................................................................
2587.2 Maintenance program ......................................................................................................................................................
2617.3 Weekly maintenance .........................................................................................................................................................
2627.3.1 Change pump rinsing solution ......................................................................................................................................................................
2667.3.2 Replace the inline filter .....................................................................................................................................................................................
2677.3.3 Clean the Fraction collector F9-C sensors ................................................................................................................................................
2697.4 Monthly maintenance .......................................................................................................................................................
4 ÄKTA pure User Manual 29119969 AB
Table of Contents
2727.5 Semiannual maintenance ...............................................................................................................................................
2737.5.1 Clean the UV flow cell ........................................................................................................................................................................................
2757.5.2 Replace the pH electrode .................................................................................................................................................................................
2767.6 Maintenance when required ..........................................................................................................................................
2777.6.1 Clean the instrument externally ...................................................................................................................................................................
2787.6.2 Perform System CIP ...........................................................................................................................................................................................
2847.6.3 Perform Column CIP ..........................................................................................................................................................................................
2887.6.4 Clean Fraction collector F9-C .........................................................................................................................................................................
2907.6.5 Clean Fraction collector F9-R .........................................................................................................................................................................
2917.6.6 Storage of the pH electrode ...........................................................................................................................................................................
2937.6.7 Clean the pH electrode .....................................................................................................................................................................................
2967.6.8 Clean the pump head check valves ............................................................................................................................................................
2997.6.9 Wipe off excess oil from the pump head ..................................................................................................................................................
3007.6.10 Clean the Conductivity flow cell ....................................................................................................................................................................
3027.7 Calibration procedures .....................................................................................................................................................
3037.7.1 Calibrate the pH monitor .................................................................................................................................................................................
3057.7.2 Calibrate the pressure monitors ...................................................................................................................................................................
3087.7.3 Calibrate the Conductivity monitor .............................................................................................................................................................
3127.7.4 Calibrate the UV monitors ...............................................................................................................................................................................
3167.8 Replacement procedures ................................................................................................................................................
3177.8.1 Replace tubing and connectors ....................................................................................................................................................................
3197.8.2 Replace internal tubing in Fraction collector F9-C ...............................................................................................................................
3227.8.3 Replace the Mixer ................................................................................................................................................................................................
3237.8.4 Replace the O-ring inside the Mixer ............................................................................................................................................................
3257.8.5 Replace the UV monitor U9-M flow cell .....................................................................................................................................................
3287.8.6 Replace the UV monitor U9-L flow cell ......................................................................................................................................................
3307.8.7 Replace Flow restrictor .....................................................................................................................................................................................
3317.8.8 Replace the inlet filters .....................................................................................................................................................................................
3327.8.9 Replace the pump head check valves ........................................................................................................................................................
3357.8.10 Replace pump piston seals of Pump P9 or P9H ....................................................................................................................................
3447.8.11 Replace pump piston seals of Pump P9-S ...............................................................................................................................................
3517.8.12 Replace pump pistons ......................................................................................................................................................................................
3527.8.13 Replace pump rinsing system tubing .........................................................................................................................................................
3558 Troubleshooting ...............................................................................................................................................
3568.1 Introduction to troubleshooting ...................................................................................................................................
3588.2 Troubleshooting: General Checklist ............................................................................................................................
3608.3 Troubleshooting: Monitors ..............................................................................................................................................
3758.4 Troubleshooting: Valves ...................................................................................................................................................
3778.5 Troubleshooting: Fraction collector ............................................................................................................................
3878.6 Troubleshooting: Pumps ..................................................................................................................................................
3948.7 Troubleshooting: Other components .........................................................................................................................
4018.8 Troubleshooting: Instrument communication .......................................................................................................
4038.9 Troubleshooting: Method development ...................................................................................................................
4048.10 Error codes .............................................................................................................................................................................
4109 Reference information ....................................................................................................................................
4119.1 System specifications ........................................................................................................................................................
4139.2 Module specifications .......................................................................................................................................................
4209.3 Tubing and connectors .....................................................................................................................................................
4289.4 Recommended tubing kits for prepacked columns ............................................................................................
4319.5 Chemical resistance guide ..............................................................................................................................................
4329.5.1 General information about biocompatibility and chemical resistance .......................................................................................
4339.5.2 Chemical resistance specifications .............................................................................................................................................................
4369.6 Wetted materials .................................................................................................................................................................
ÄKTA pure User Manual 29119969 AB 5
Table of Contents
4389.7 Predefined methods and phases .................................................................................................................................
4399.7.1 Predefined purification methods ..................................................................................................................................................................
4459.7.2 Predefined maintenance methods ..............................................................................................................................................................
4479.7.3 Predefined phases ..............................................................................................................................................................................................
4499.8 System settings ....................................................................................................................................................................
4509.8.1 System settings - UV ..........................................................................................................................................................................................
4519.8.2 System settings - Conductivity ......................................................................................................................................................................
4529.8.3 System settings - pH ..........................................................................................................................................................................................
4539.8.4 System settings - Pressure alarms ..............................................................................................................................................................
4559.8.5 System settings - Air sensor ...........................................................................................................................................................................
4579.8.6 System settings - I/O-box ................................................................................................................................................................................
4589.8.7 System settings - Fraction collection ..........................................................................................................................................................
4609.8.8 System settings - Tubing and Delay volumes .........................................................................................................................................
4629.8.9 System settings - Wash settings ..................................................................................................................................................................
4639.8.10 System settings - Watch parameters ........................................................................................................................................................
4649.8.11 System settings - Advanced ...........................................................................................................................................................................
4659.8.12 System settings - Data collection .................................................................................................................................................................
4669.9 Manual instructions ...........................................................................................................................................................
4679.9.1 Manual instructions - Pumps .........................................................................................................................................................................
4719.9.2 Manual instructions - Flow path ..................................................................................................................................................................
4739.9.3 Manual instructions - Monitors .....................................................................................................................................................................
4749.9.4 Manual instructions - Fraction collection .................................................................................................................................................
4789.9.5 Manual instructions - I/O-box ........................................................................................................................................................................
4799.9.6 Manual instructions - Alarms .........................................................................................................................................................................
4829.9.7 Manual instructions - Wash settings ..........................................................................................................................................................
4839.9.8 Manual instructions - Watch parameters ................................................................................................................................................
4849.9.9 Manual instructions - Advanced ...................................................................................................................................................................
4879.9.10 Manual instructions - Other ............................................................................................................................................................................
4889.10 Available Run data ..............................................................................................................................................................
4919.11 Available Curves ..................................................................................................................................................................
4939.12 Injection volumes and peak broadening ..................................................................................................................
4949.13 Delay volumes ......................................................................................................................................................................
4999.14 Component volumes .........................................................................................................................................................
5019.15 Pressure control ...................................................................................................................................................................
5039.16 Node IDs ..................................................................................................................................................................................
50610 Ordering information ......................................................................................................................................
514Index ...................................................................................................................................................................
6 ÄKTA pure User Manual 29119969 AB
1 Introduction
Purpose of the User Manual
The User Manual provides you with instructions and information to run the ÄKTA pure system. It also
includes relevant guidance for practical handling and maintenance of instrument components.
In this chapter
This chapter contains the following sections:
1 Introduction
See pageSection
81.1 Important user information
101.2 ÄKTA pure overview
121.3 ÄKTA pure user documentation
ÄKTA pure User Manual 29119969 AB 7
1 Introduction
1.1 Important user information
1.1 Important user information
Read this before operating
ÄKTA pure
All users must read the entire ÄKTA pure Operating Instructions before installing, operating, or
maintaining the instrument. Always keepthe ÄKTA pure OperatingInstructions at hand whenoperating
ÄKTA pure.
Do not operate ÄKTA pure in any other way than described in the user documentation. If you do, you
may be exposed to hazards that can lead to personal injury and you may cause damage to the
equipment.
Intended use
ÄKTA pure is intended for purification of bio-molecules, in particular proteins, for research purposes
by trained laboratory staff members in research laboratories.
ÄKTA pure shall not be used in any clinical procedures, or for diagnostic purposes.
Prerequisites
In order to operate the system according to the intended purpose, it is important that:
•
•
•
•
Safety notices
This user documentation contains safety notices (WARNING, CAUTION, and NOTICE) concerning the
safe use of the product. See definitions below.
you have a general understanding of how the computer and the Microsoft®Windows®operating
system work.
you understand the concepts of liquid chromatography.
you have read and understood the Safety instructions chapter in ÄKTA pure Operating Instructions.
a user account has been created according to UNICORN™ Administration and Technical Manual.
WARNING
WARNINGindicates a hazardous situationwhich,if not avoided, could resultin death
or serious injury. It is important not to proceed until all stated conditions are met
and clearly understood.
8 ÄKTA pure User Manual 29119969 AB
Notes and tips
1 Introduction
1.1 Important user information
CAUTION
CAUTION indicatesa hazardous situationwhich, if not avoided,could result inminor
or moderate injury. It is important not to proceed until all stated conditions are met
and clearly understood.
NOTICE
NOTICE indicatesinstructionsthat must befollowed to avoid damageto the product
or other equipment.
Note:
Tip:
A Note is used to indicate information that is important for trouble-free and optimal use of the
product.
A tip contains useful information that can improve or optimize your procedures.
ÄKTA pure User Manual 29119969 AB 9
1 Introduction
1.2 ÄKTA pure overview
1.2 ÄKTA pure overview
Introduction
ÄKTA pure is intended for purification of bio-molecules, in particular proteins, for research purposes
by trained laboratory staff members in research laboratories.
This section gives an overview of the ÄKTA pure instrument and the UNICORN software. For detailed
information about UNICORN, see the UNICORN manuals listed in UNICORN user documentation, on
page 13. For detailed information about the instrument, see Chapter 2 The ÄKTA pure instrument, on
page 14.
Main features
The main features of ÄKTA pure are listed below.
•
ÄKTA pure is a flexible system that allows the user to configure both hardware and software to
meet many purification needs.
•
The instrument can be easily extended with additional valves, detectors and fraction collectors.
•
There are a large number of different hardware modules to choose from. The user can adjust, for
example, the number of columns, inlets, outlets and detectors and choose between different ways
to apply and collect samples.
•
Purification and maintenance methods are easily created using predefined methods and method
phases. Amethod phase refers toaspecific step/part inthe method, suchas column wash orelution.
Method phasesare described inPhaseProperties and are displayedgraphically in amethod outline.
This makes methods and phases easy to understand and edit.
•
ÄKTA pure is controlled by the UNICORN software: a complete package for control, supervision and
evaluation of chromatography instruments and purification runs.
•
UNICORN has different software licence options, such as Design of Experiments (DoE) and Column
logbook, to further add user support.
UNICORN modules overview
UNICORN consists of four modules: Administration, Method Editor, System Control and Evaluation.
The main functions of each module are described in the following table.
Main functionsModule
Administration
10 ÄKTA pure User Manual 29119969 AB
Perform user and system setup, system log and database administration.
1 Introduction
1.2 ÄKTA pure overview
Main functionsModule
Create and edit methods using one or a combination of:Method Editor
•
Predefined methods with built-in application support
•
Drag-and-drop function to build methods with relevant steps
•
Line-by-line text editing
The interface provides easy viewing and editing of run properties.
System Control
Start, monitor and control runs. The current flow path is illustrated
in the Process Picture, which allows manual interactions with the
system and provides feedback on run parameters.
Open results, evaluate runs and create reports.Evaluation
The default Evaluation module includes a user interface optimized
for workflows like quick evaluation, compare results and work with
peaks and fractions.
To perform operations like Design of Experiments, users can easily
switch to Evaluation Classic.
When working with the modules Administration, Method Editor, System Control and Evaluation it is
possible to access descriptions of the active window by pressing the F1 key. This can be especially
helpful when editing methods
ÄKTA pure User Manual 29119969 AB 11
1 Introduction
1.3 ÄKTA pure user documentation
1.3 ÄKTA pure user documentation
Introduction
This section describes the user documentation that is delivered with ÄKTA pure.
User documentation
The user documentation listed in the table below is delivered with ÄKTA pure. It is also available on the
user documentation CD.
Main contentsDocument
ÄKTA pure Operating Instructions
Fraction collector F9-C and F9-R
Operating Instructions
Sample pump S9 and S9H Operating
Instructions
ÄKTA pure User Manual
Instructions needed to install, operate and maintain ÄKTA
pure in a safe way.
Instructions neededto install, operateand maintain Fraction
collectors F9-C and F9-R in a safe way.
Instructions neededto install, operate andmaintainSample
pump S9 and S9H in a safe way.
Detailed instrument and module descriptions and instructions on how torun, maintain and troubleshootthe system.
12 ÄKTA pure User Manual 29119969 AB
UNICORN user documentation
The user documentation listed in the following table is available from the Help menu in UNICORN or
from the UNICORN Online Help and Documentation software accessed by pressing the F1 key in any
UNICORN module.
1 Introduction
1.3 ÄKTA pure user documentation
Main contentsDocumentation
UNICORN Help
Getting started with Evaluation
Note:
Available in UNICORN 7.0 and later.
UNICORN Method Manual
1
UNICORN Administration and
Technical Manual
UNICORN Evaluation Manual
1
1
UNICORN System Control Manual
Descriptions ofUNICORNdialog boxes (availablefrom the Help
menu).
•
Video clips showing common workflows in the Evaluation
module.
•
Overview of features of the Evaluation module.
•
Overview and detailed descriptions of the method creation
features in UNICORN.
•
Workflow descriptions for common operations.
•
Overview and detailed description of network setup and
complete software installation.
•
Administration of UNICORN and the UNICORN database.
•
Overview and detaileddescriptionsof the Evaluation Classic
module in UNICORN.
•
Description of the evaluation algorithms used in UNICORN.
1
•
Overview and detailed description of the system control
features in UNICORN.
•
Includes generaloperation, system settings andinstructions
on how to perform a run.
1
Current UNICORN version is added to the title of the manual.
Additional literature
For practicaltips on chromatography, refer to ÄKTA Laboratory-scale:Chromatography Systems Instrument Management Handbook (product code 29010831).
ÄKTA pure User Manual 29119969 AB 13
2 The ÄKTA pure instrument
2 The ÄKTA pure instrument
About this chapter
This chapterprovides an overview of theÄKTA pureinstrument. It alsodescribes the internalinstrument
components and how these are installed in the instrument.
In this chapter
This chapter contains the following sections:
See pageSection
152.1 Overview illustrations
262.2 Liquid flow path
282.3 Instrument control panel
332.4 Instrument modules
772.5 Installation of internal modules
802.6 Accessories
14 ÄKTA pure User Manual 29119969 AB
2.1 Overview illustrations
1 2
4
3
5
6
Introduction
This section provides an overview of the system and its available modules.
Core module configurations
ÄKTA pure is available with two core module configurations, one for flow rates up to 25 ml/min and
one for flow rates up to 150 ml/min. In this manual they are referred to as ÄKTA pure 25 (25 ml/min)
and ÄKTA pure 150 (150 ml/min).
Illustrations of the main parts of the instrument
The illustrations below show the location of the main parts of the instrument.
2 The ÄKTA pure instrument
2.1 Overview illustrations
ÄKTA pure User Manual 29119969 AB 15
FunctionPart
Wet sides1
Buffer tray2
Holder rails3
Instrument control panel4
Power switch5
Ventilation panel6
5
6
7
8
9
10
11 12
2
3
4
1
2 The ÄKTA pure instrument
2.1 Overview illustrations
Example of a typical configuration of the wet side
A typical configuration of ÄKTA pure is illustrated below.
FunctionPart
Multi-module panel1
Inlet valve2
Pump rinsing liquid tube3
System pump B4
Pressure monitor5
System pump A6
Mixer7
Outlet valve8
Injection valve9
Conductivity monitor10
Column valve11
UV monitor12
16 ÄKTA pure User Manual 29119969 AB
Available modules
The modular design allows the user to customize ÄKTA pure in multiple ways. The system is always
delivered with the core modules of the selected configuration, but optional modules may be added to
the flow path.
The table below lists the available modules forÄKTA pure 25 and ÄKTA pure 150. Core modules are indicated with an asterisk (*).
2 The ÄKTA pure instrument
2.1 Overview illustrations
Note:
The valves for ÄKTA pure 25 and ÄKTA pure 150 are compatible with both systems but for the
best performance the specific valve type should be used. The narrow channels in the valves
for ÄKTA pure 25 will give too high back pressure if used above 50 ml/min. The larger volumes
in the “H” valves for ÄKTA pure 150 may decrease resolution and increase peak broadening if
used in ÄKTA pure 25.
Label inModule
ÄKTA pure 150ÄKTA pure 25
P9H AP9 ASystem pump A*
P9H BP9 BSystem pump B*
R9R9Pressure monitor*
M9M9Mixer*
V9H-InjV9-InjInjection valve*
V9H-IAV9-IAInlet valve A
V9H-IBV9-IBInlet valve B
V9H-IABV9-IABInlet valve AB
V9H-IXV9-IXInlet valve IX
V9H-ISV9-ISSample inlet valve
V9H-MV9-MMixer valve
V9H-LV9-LLoop valve
V9H-CV9-CColumn valves
V9H-CsV9-Cs
V9H-pHV9-pHpH valve
V9H-OV9-OOutlet valves
V9H-OsV9-Os
V9H-VV9-VVersatile valve
U9-LU9-LUV monitors
U9-MU9-M
ÄKTA pure User Manual 29119969 AB 17
3E
3F
V9-L
1
E
2
E
4
E
5
E
5
F
4
F
2
F
1
F
E
F
2 The ÄKTA pure instrument
2.1 Overview illustrations
Illustration convention
Label inModule
ÄKTA pure 150ÄKTA pure 25
C9C9Conductivity monitor
L9-1.5L9-1.5External air sensor
L9-1.2L9-1.2
F9-CF9-CFraction collectors
F9-RF9-R
E9E9I/O-box
S9HS9Sample pump
In the valve illustrations below, the following convention is used to point out the location of the ports
on the valve head. Loop valve V9-L is used as an example.
Core modules
•
Ports located on the valve head rim are indicated outside the black
ring (e.g., 1E, 2E, etc.).
•
Ports located on the pivot part of the valve head are indicated on
the inside of the black ring (e.g., 3E and 3F).
•
Ports located on the valve head front are indicated by an arrow
(e.g., E and F).
DescriptionCore module
A high precision pump, which delivers buffer or sample in purification runs.System pump P9 A or P9H A
For further information, refer to Section 2.4.1 System pumps, on page 34.
18 ÄKTA pure User Manual 29119969 AB
L
o
o
p
E
L
o
o
p
F
S
y
r
W
1
P
a
S
2
W
V9-Inj
2 The ÄKTA pure instrument
2.1 Overview illustrations
DescriptionCore module
A high precision pump, which delivers buffer in purification runs.System pump P9 B or P9H B
For further information, refer to Section 2.4.1 System pumps, on page 34.
Reads the system pressure after System pump A and System pump B.Pressure monitor R9
For furtherinformation, refer to Section 2.4.12 Pressure monitors, on page 68.
Mixer M9
Mixes thebuffers deliveredfrom thesystempumps to a homogeneousbuffer
composition.
Three Mixer chambers are available for ÄKTA pure 25, their volumes are: 0.6
ml, 1.4 ml (mounted at delivery) and 5 ml.
Three Mixer chambers are available for ÄKTA pure 150. Their volumes are:
1.4 ml (mounted at delivery), 5 ml (included in delivery), and 15 ml.
CAUTION
Risk of explosion. Do not use Mixer chamber 15 ml with
an ÄKTA pure 25 system configuration. The maximum
pressure for Mixer chamber 15 ml is 5 MPa.
For further information, refer to Section 2.4.2 Mixer, on page 38.
Directs sample onto the column.Injection valve V9-Inj or V9H-Inj
For further information, refer to Section 2.4.6 Injection valve, on page 50.
ÄKTA pure User Manual 29119969 AB 19
V9-IA
Q
A1
A2
A3
A4
A7
A6
A5
Out
V9-IB
B5
B4
B3
B2
B1
B6
B7
Q
Out
A
B
A1
A2
B1
B2
V9-IS
S5
S4
S3
S2
S1
S6
S7
Buff
Out
2 The ÄKTA pure instrument
2.1 Overview illustrations
Optional modules
DescriptionModule
Inlet valve for SystempumpA with seveninlet ports andintegrated air sensor.Inlet valve V9-IA or V9H-IA
For further information, refer to Section 2.4.4 Inlet valves, on page 40.
Inlet valve for SystempumpB with seveninlet ports andintegrated air sensor.Inlet valve V9-IB or V9H-IB
For further information, refer to Section 2.4.4 Inlet valves, on page 40.
Inlet valve V9-IAB or V9H-IAB
Inlet valve with two A inlet ports and two B inlet ports. No integrated air
sensor.
For further information, refer to Section 2.4.4 Inlet valves, on page 40.
Sample inlet valve V9-IS or V9H-IS
Inlet valve with eight inlet ports (seven sample inlets and one buffer inlet)
and integratedair sensor. The Sampleinletvalve requiresan external Sample
pump module.
For further information, refer to Section 2.4.4 Inlet valves, on page 40.
20 ÄKTA pure User Manual 29119969 AB
V9-IX
5
4
3
2
1
6
7
8
Out
V9-M
Re-Inj
Out
In
Mixer
2 The ÄKTA pure instrument
2.1 Overview illustrations
DescriptionModule
Inlet valve with eight inlet ports. No integrated air sensor.Inlet valve V9-IX or V9H-IX
For further information, refer to Section 2.4.4 Inlet valves, on page 40.
Mixer valve V9-M or V9H-M
Directs the flowto the Injection valve,bypassing the Mixer, orto the Injection
valve via the Mixer.
For further information, refer to Section 2.4.5 Mixer valve, on page 47.
Enables the use of up to five loops connected to the instrument.Loop valve V9-L or V9H-L
For further information, refer to Section 2.4.7 Loop valve, on page 53.
Column valve V9-C or V9H-C
Connects up to five columns to the instrument, and directs the flow to one
column ata time. The Columnvalve featurestwointegrated pressuresensors.
Allows the user to choose flow direction through the column, or to bypass
the column.
For further information, refer to Section 2.4.8 Column valves, on page 56.
ÄKTA pure User Manual 29119969 AB 21
V9-Cs
A
Out
In
B
V9-O
Out2
Out1
Frac
Out10
In
Out4
W
Out6
Out3
Out9
Out5
Out7
Out8
V9-Os
In
Frac
Out
W
2 The ÄKTA pure instrument
2.1 Overview illustrations
DescriptionModule
Connects a single column to the instrument.Column valve V9-Cs or V9H-Cs
Allows theuser to chose flow direction throughthecolumn,or to bypass the
column.
For further information, refer to Section 2.4.8 Column valves, on page 56.
pH valve V9-pH or V9H-pH
Outlet valve V9-O or V9H-O
Enables the pH electrodetobe included in the flow path or bypassed during
a run. The pH electrode may be calibrated when installed in the pH valve.
For further information, refer to Section 2.4.10 pH valve, on page 61.
Directs the flow to the Fraction collector, Fraction collector 2 (out 10), any
of the ten outlet ports, or waste.
For further information, refer to Section 2.4.11 Outlet valves, on page 65.
Outlet valve V9-Os or V9H-Os
Directs the flow to theFraction collector, Fraction collector 2, theoutlet port,
or waste.
For further information, refer to Section 2.4.11 Outlet valves, on page 65.
22 ÄKTA pure User Manual 29119969 AB
V9-V
1
4
2
3
2 The ÄKTA pure instrument
2.1 Overview illustrations
DescriptionModule
Versatile valve V9-V or V9H-V
A 4-port, 4-position valve, which can be used when adding extra features
to the flow path.
For further information, refer to Section 2.4.9 Versatile valve, on page 60.
Measures the UV absorbance at a fixed wavelength of 280 nm.UV monitor U9-L
For further information, refer to Section 2.4.13 UV monitors, on page 70.
UV monitor U9-M
Measures the UV/Visabsorbanceat up to threewavelengths simultaneously
in the range 190-700 nm.
For further information, refer to Section 2.4.13 UV monitors, on page 70.
Measures the conductivity of buffers and eluted proteins.Conductivity monitor C9
For further information, refer to Section 2.4.14 Conductivity monitor, on
page 73.
ÄKTA pure User Manual 29119969 AB 23
2 The ÄKTA pure instrument
2.1 Overview illustrations
DescriptionModule
Prevents air from being introduced into the flow path.External air sensor L9-1.5 or L9-1.2
For further information, refer to Section 3.1 External air sensors, on page 88.
Flexible fraction collector that can collect up to 576 fractions.Fraction collector F9-C
Up to two fraction collectors can be connected at the same time, of which
only one (the primary) can be a Fraction collector F9-C.
I/O-box E9
For furtherinformation, refer to Section 3.2 Fractioncollector F9-C, on page 90.
Round fraction collector that can collect up to 175 fractions.Fraction collector F9-R
Up to two fraction collectors can be connected at the same time.
For further information, refer to Section 3.3 Fraction collector F9-R , on
page 104.
Receives analog or digital signals from, or transfers analog or digitalsignals
to, external equipment that has been incorporated in the system.
For further information, refer to Section 3.5 I/O-box E9, on page 112.
24 ÄKTA pure User Manual 29119969 AB
2 The ÄKTA pure instrument
2.1 Overview illustrations
DescriptionModule
Sample pump S9 or S9H
A high precision pump with an integrated pressure monitor. The sample
pump delivers buffer or sample in purification runs.
For further information, refer to Section 3.4 Sample pump S9 and S9H, on
page 108.
ÄKTA pure User Manual 29119969 AB 25
4
5
6
7
8
9
10
11
12
13
14
15
16
17
W1 W2
W
3
2
1
2 The ÄKTA pure instrument
2.2 Liquid flow path
2.2 Liquid flow path
Introduction
ÄKTA pure is a liquid chromatography system with a flexible flow path.
This section provides an overview of the liquid flow path, and its possibilities.
Example of a typical liquid flow path
The illustration belowshows the flow pathfora typical system configuration.The individual instrument
modules are presented in the table below. The configuration of the system is defined by the user.
DescriptionPart
Pressure monitor1
Sample pump2
Sample inlet valve3
26 ÄKTA pure User Manual 29119969 AB
2 The ÄKTA pure instrument
2.2 Liquid flow path
DescriptionPart
Inlet valve4
System pump B5
System pump A6
Pressure monitor7
Mixer8
Injection valve9
Sample loop or Superloop10
Column valve11
Column12
UV monitor13
Conductivity monitor14
Flow restrictor15
Outlet valve16
Fraction collector17
WasteW, W1, W2
ÄKTA pure User Manual 29119969 AB 27
1
2
3
4
2 The ÄKTA pure instrument
2.3 Instrument control panel
2.3 Instrument control panel
Introduction
This section describes the design and main function of the Instrument control panel B9.
Function of the Instrument control panel
The Instrument control panel shows the current state of the system. The Pause and Continue buttons
can be used to control an ongoing run. It is possible to lock and unlock the Instrument control buttons
from UNICORN.
Location and illustration
The illustration below shows the location and detailed view of the Instrument control panel.
FunctionPart
Power/Communication indicator (white)1
Continue button with a green light indicator2
Pause button with an orange light indicator3
Alarm and error indicator (red)4
28 ÄKTA pure User Manual 29119969 AB
Lock/Unlock function
Follow the instruction below to lock or unlock the Pause and Continue buttons of the Instrument
control panel from UNICORN.
ActionStep
In System Control, select System:Settings.1
Result: The System Settings dialog opens.
In the System Settings dialog:2
•
Select Advanced:Instrument control panel.
•
Select Locked or Unlocked.
•
Click OK.
2 The ÄKTA pure instrument
2.3 Instrument control panel
Buttons
The Instrument control panel includes the following buttons:
FunctionPart
Resumes instrument operation from the following states:
•
Pause
•
Wash
•
Hold
Pauses the run and stops all pumps.
ÄKTA pure User Manual 29119969 AB 29
2 The ÄKTA pure instrument
2.3 Instrument control panel
Status indications
The light indicators on the Instrument control panel indicate the current status of ÄKTA pure.
The table below describes the different states that can be displayed.
DescriptionStateDisplay
The instrument is turned off.OffAll light indicators are off.
flashes slowly.
flashes quickly.
displays a constant light.
Power-onThe Power/Communicationindicator
The instrumenthas no communication
with the Instrument server.
The system is starting up.ConnectingThe Power/Communicationindicator
The instrument is ready to use.ReadyThePower/Communication indicator
30 ÄKTA pure User Manual 29119969 AB
cator and Continue button display a
constant light.
2 The ÄKTA pure instrument
2.3 Instrument control panel
DescriptionStateDisplay
A run is ongoing.RunBoth the Power/Communicationindi-
displays aconstantlightand the Con-
tinue button flashes slowly.
cator and Pause button display a
constant light.
The Power/Communicationindicator
displays a constant light. and the
Alarm and error indicator flashes.
WashThe Power/Communicationindicator
Alarms and errors
A wash instructionora pump synchronization is ongoing.
A run has been put on hold.Hold
A run has been paused.PauseBoth the Power/Communicationindi-
The system has been paused due to
an alarm.To resumethe run, acknowledge the alarm and continue the run
in UNICORN.
ÄKTA pure User Manual 29119969 AB 31
2 The ÄKTA pure instrument
2.3 Instrument control panel
displays a pulsating light.
DescriptionStateDisplay
The system is in power-saving mode.Power-saveThe Power/Communicationindicator
Re-programmingAll indicators are lit in a wave pattern.
A module is being re-programmed to
be compatiblewith the current instrument configuration.
32 ÄKTA pure User Manual 29119969 AB
2.4 Instrument modules
Introduction
This section describes the design and main functions of the instrument modules.
In this section
This section contains the following subsections:
2 The ÄKTA pure instrument
2.4 Instrument modules
See pageSection
342.4.1 System pumps
382.4.2 Mixer
392.4.3 Valves, overview
402.4.4 Inlet valves
472.4.5 Mixer valve
502.4.6 Injection valve
532.4.7 Loop valve
562.4.8 Column valves
602.4.9 Versatile valve
612.4.10 pH valve
652.4.11 Outlet valves
682.4.12 Pressure monitors
702.4.13 UV monitors
732.4.14 Conductivity monitor
752.4.15 Flow restrictor
ÄKTA pure User Manual 29119969 AB 33
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.1 System pumps
2.4.1 System pumps
Introduction
This section describes the design and main functions of the system pumps, and also the pump piston
rinsing systems. The system can also be equipped with an external, optional sample pump, see Sec-
tion 3.4 Sample pump S9 and S9H, on page 108.
Function of the system pumps
The ÄKTA pure instrument is fitted with two high precision system pumps, System pump A andSystem
pump B.The system pumpscan be used individually,orin combination togenerate isocratic or gradient
elution in purification methods.
Each pumpmodule consists oftwopump heads thatworkalternately to givea continuous, lowpulsation,
liquid delivery. Toensure delivery ofcorrect liquid volume,thepumps must befree fromair. Each pump
head is equippedwitha purge valvethat is usedforthis purpose. See Section 5.4Prime inlets and purge
pump heads, on page 161.
The table below contains the operating limits and labels of the system pumps of ÄKTA pure 25 and
ÄKTA pure 150, respectively.
Max. pressureFlow ratePump typeLabelConfiguration
P9P9 A and P9 BÄKTA pure 25
Note:
When running the
Column packing
flow instruction, the
maximum flow rate
is 50 ml/min.
P9HP9H A and P9H BÄKTA pure 150
Note:
When running the
Column packing
flow instruction, the
maximum flow rate
is 300 ml/min.
20 MPa0.001 – 25 ml/min
5 MPa0.01 – 150 ml/min
34 ÄKTA pure User Manual 29119969 AB
Location and illustration
1 2 3
45678
The illustrationbelow shows thelocationof System pumpA and System pumpB,together with a detailed
view of a system pump.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.1 System pumps
DescriptionPart
Purge valve: Used to remove air from the pump1
Outlet port with check valve2
3
Connections to pump piston rinsing system: Tubing is connected between the pumps
and the Pump piston rinsing system tube (6)
Inlet port with check valve4
Pump head: Encapsulates the inner parts of the pump5
Pump piston rinsing system tube6
System pump B7
System pump A8
The pump piston rinsing system
A sealprevents leakage betweenthe pump chamber andthe drive mechanism.The seal is continuously
lubricated by the presence of liquid. The pump piston rinsing system continuously flushes the low
pressure chamber behind the piston with a low flow of 20% ethanol. This prevents any deposition of
salts from aqueous buffers on the pistons and prolongs the working life of the seals.
The pump piston rinsing system tubing is connected to the rearmost holes on the pump heads.
For instructions on how to fill the rinsing system, see Prime the system pump piston rinsing system, on
page 263.
ÄKTA pure User Manual 29119969 AB 35
1
2
3
4
5
1
2
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.1 System pumps
Illustration of the pump piston rinsing system
DescriptionPart
Rinsing system tube holder, top1
Rinsing system tube2
Rinsing system tube holder, bottom3
Outlet tubing4
Inlet tubing5
System pump rinsing systems flow path
The illustration below shows the tubing configuration of the pumppiston rinsing system of the system
pumps.
36 ÄKTA pure User Manual 29119969 AB
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.1 System pumps
DescriptionPart
System pump A1
System pump B2
ÄKTA pure User Manual 29119969 AB 37
1
2
3
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.2 Mixer
2.4.2 Mixer
Function of the Mixer
Mixer M9 is located after System pump A and System pump B, and before the Injection valve. The
Mixer is a dynamic mixer for high-performance gradients. It is used to make surethat the buffers from
the System pumps are mixed to give a homogenous buffer composition.
The Mixer has a built-in filter that prevents particles from reaching the column.
Note:
Replace the sealing ring of the mixer with a highly resistant O-ring if the system is going to be
exposed to organic solvents or high concentrations of organic acids for longer periods of time.
Refer to Section 7.8.4 Replace the O-ring inside the Mixer, on page 323 for how to change the
ring.
Location and illustration
The illustration below shows the location, together with a detailed view of Mixer M9.
DescriptionPart
Outlet1
Mixer chamber (0.6, 1.4, 5 or 15 ml)2
CAUTION
Risk of explosion. Do not use Mixer chamber 15 ml with
Inlet3
38 ÄKTA pure User Manual 29119969 AB
an ÄKTA pure 25 configuration. The maximum pressure
for Mixer chamber 15 ml is 5 MPa.
2.4.3 Valves, overview
General design and function of rotary valves
The valves of the ÄKTA pure instrument allow flexibility in the liquid flow path.
All valves used in the ÄKTA pure instrument are rotary valves. The motorized rotary valve consists of
a Valve connection block with a number of defined bores with channels to the inlet and outlet ports
of the valve. The Rotary disc, mounted on the motor, has a number of defined channels. The pattern
of channelsof the Rotarydisc together withthe pattern and locationofthe ports oftheValve connection
block, define the flow path and function of each type of valve. When the Rotary disc turns, the flow
path in the valve changes.
Illustration of inlet valve components
The illustration below shows the components of a disassembled Inlet valve A or a disassembled Inlet
valve B.
Inlet valve AB is built up by the same parts but have another configuration of defined bores.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.3 Valves, overview
DescriptionPart
Valve connection block (stator)1
Rotary disc (rotor)2
Defined channel(s) in the Rotary disc3
Defined bores in the Valve connection block4
Note:
ÄKTA pure User Manual 29119969 AB 39
Inlet and outlet ports are not visible in the picture. They are located on the opposite side of the
Valve connection block.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.4 Inlet valves
2.4.4 Inlet valves
Function of the inlet valves
The inlet valves are used to select which buffers or samples to use in a run. The inlet valves available
for ÄKTA pure and their functions are described in the table below.
FunctionLabel inInlet valve
ÄKTA pure 150ÄKTA pure 25
V9H-IAV9-IAInlet valve A
Enables automaticchange between different buffers
and wash solutions (seven inlet ports) .
Can be used together with Inlet valve B to generate
gradients by mixing bufferfrom System pump A and
buffer from System pump B.
V9H-IBV9-IBInlet valve B
Enables automaticchange between different buffers
and wash solutions (seven inlet ports).
Can be used together with Inlet valve A to generate
gradients by mixing bufferfrom System pump A and
buffer from System pump B.
V9H-IABV9-IABInlet valve AB
Enables automaticchange between different buffers
and wash solutions (two A and two B inlet ports).
Can be used to generate gradients by mixing buffer
from System pump A and buffer from System pump
B.
V9H-ISV9-ISSample inletvalve
Enables automatic loading of up to seven samples
when used together with a sample pump.
Increases the total number of inlets to the system.V9H-IXV9-IXInlet valve X
Can be used in two different configurations X1 and
X2. The configurations are called V9-X1 and V9-X2
for ÄKTA pure 25 and, V9H-X1 and V9H-X2 for ÄKTA
pure 150.
The modular design of ÄKTA pure allows the use of several combinations of inlet valves.
The possible combinations of Inlet valve A, Inlet valve B and Inlet valve AB are:
•
one Inlet valve A
•
one Inlet valve B
•
Inlet valve A and Inlet valve B,
•
Inlet valve AB together with Inlet valve A or Inlet valve B,
•
one Inlet valve AB,
or
40 ÄKTA pure User Manual 29119969 AB
•
1 2
no installed inlet valves.
The sample inlet valve can be used together with any of the combinations listed above.
The air sensors integrated in Inlet valve A, Inlet valve B, and Sample inlet valve detect the presence of
air and prevent the air from entering the pump.
Inlet valve AB and Inlet valve IX lack built-in air sensors, but can be used together with external air
sensors.
Location of inlet valves
The locations of the inlet valves are described in the following table:
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.4 Inlet valves
LocationInlet valve
Before System pump AInlet valve A
Before System pump BInlet valve B
Before both System pump A and System pump BInlet valve AB
Illustration of Inlet valve A and Inlet valve B
The illustrationbelow shows adetailed view ofInletvalve A andInlet valve B, in this examplewithlabels
V9-IA and V9-IB.
DescriptionPart
Integrated air sensor of Inlet valve A (located under the plug)1
Before the sample pumpSample inlet valve
For example, before another inlet valveInlet valve IX
Integrated air sensor of Inlet valve B (located under the plug)2
ÄKTA pure User Manual 29119969 AB 41
Out
Out
1 2
34
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.4 Inlet valves
Ports of Inlet valve A and Inlet valve B
The illustration below shows the ports of Inlet valve A and Inlet valve B, in this example with labels V9IA and V9-IB.
DescriptionPort
Illustration of Inlet valve AB
The illustration below shows a detailed view of Inlet valve AB.
Buffer inlets of Inlet valve AA1-A7
Buffer inlets of Inlet valve BB1-B7
Not used for ÄKTA pureQ
To the respective System pumpOut
42 ÄKTA pure User Manual 29119969 AB
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.4 Inlet valves
DescriptionPart
A inlet ports1
B inlet ports2
Outlet port to System pump B3
Outlet port to System pump A4
Note:
Inlet valve AB does not have any integrated air sensor.
Ports of Inlet valve AB
The illustration below shows the ports of Inlet valve AB.
A1-A2
DescriptionPort
Inlet ports A1 and A2 of Inlet valve AB are used when buffers or samples
should be delivered to System pump A
B1-B2
Inlet ports B1 and B2 of Inlet valve AB are used when buffers or samples
should be delivered to System pump B
Outlet port A of Inlet valve AB leads to System pump AA
Outlet port B of Inlet valve AB leads to System pump BB
ÄKTA pure User Manual 29119969 AB 43
1
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.4 Inlet valves
Illustration of Sample inlet valve
The illustration below shows a detailed view of the Sample inlet valve.
DescriptionPart
Integrated air sensor (located under the plug)1
Ports of Sample inlet valve
The illustration below shows the ports of Sample inlet valve, in this example labeled V9-IS.
DescriptionPort
Sample inletsS1-S7
Buffer inletBuff
To Sample pumpOut
44 ÄKTA pure User Manual 29119969 AB
Ports of Inlet valve IX
The illustration below shows the ports of Inlet valve IX.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.4 Inlet valves
DescriptionPort
Note:
Connect tubing
The table below shows the tubing and connectors that is delivered together with the optional inlet
valves.
label
A1-A2
and
B1-B2
A1-A7
Inlets to
Inlet valve AB
Inlets to
Inlet valve A
Inlets1-8
For example, to another inlet valveOut
Inlet valve IX does not have an integrated air sensor.
FEP, o.d. 1/8",
i.d. 1.6 mm
FEP, o.d. 3/16",
i.d. 2.9 mm
Tubing connector, 5/16" with
Ferrule (yellow),
1/8"
FEP, o.d. 1/8",
i.d. 1.6 mm
FEP, o.d. 3/16",
i.d. 2.9 mm
Tubing connector, 5/16" with
Ferrule (yellow),
1/8"
ConnectorTubingConnectionTubing
Tubing
length
ÄKTA pure 150ÄKTA pure 25ÄKTA pure 150ÄKTA pure 25
(mm)
1500Tubing connector, 5/16" with
Ferrule (blue),
3/16"
1500Tubing connector, 5/16" with
Ferrule (blue),
3/16"
B1-B7
Inlets to
Inlet valve B
ÄKTA pure User Manual 29119969 AB 45
FEP, o.d. 1/8",
i.d. 1.6 mm
FEP, o.d. 3/16",
i.d. 2.9 mm
Tubing connector, 5/16" with
Ferrule (yellow),
1/8"
tor, 5/16" with
Ferrule (blue),
3/16"
1500Tubing connec-
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.4 Inlet valves
label
InA
InB
S1-S7
Buff
From Inlet valve
A or
Inlet valve AB to
System pump A
From Inlet valve
B or
Inlet valve AB to
System pump B
Sample inlets to
Sample inlet
valve
Buffer inlet to
Sample inlet
valve
FEP, o.d. 1/8",
i.d. 1.6 mm
FEP, o.d. 1/8",
i.d. 1.6 mm
FEP o.d. 1/8",
i.d. 1.6 mm
FEP o.d. 1/8",
i.d. 1.6 mm
FEP, o.d. 3/16",
i.d. 2.9 mm
FEP, o.d. 3/16",
i.d. 2.9 mm
FEP, o.d. 3/16",
i.d. 2.9 mm
FEP, o.d. 3/16",
i.d. 2.9 mm
ConnectorTubingConnectionTubing
Tubing connector, 5/16" with
Ferrule (yellow),
1/8"
Tubing connector, 5/16" with
Ferrule (yellow),
1/8"
Tubing connector, 5/16" with
Ferrule (yellow),
1/8"
Tubing connector, 5/16" with
Ferrule (yellow),
1/8"
ÄKTA pure 150ÄKTA pure 25ÄKTA pure 150ÄKTA pure 25
tor, 5/16" with
Ferrule (blue),
3/16"
tor, 5/16" with
Ferrule (blue),
3/16"
tor, 5/16" with
Ferrule (blue),
3/16"
tor, 5/16" with
Ferrule (blue),
3/16"
Tubing
length
(mm)
300Tubing connec-
300Tubing connec-
1000Tubing connec-
1000Tubing connec-
InS
Sample inlet
valve to
Sample Pump
Note:
FEP o.d. 1/8",
i.d. 1.6 mm
FEP, o.d. 3/16",
i.d. 2.9 mm
Tubing connector, 5/16" with
Ferrule (yellow),
1/8"
tor, 5/16" with
Ferrule (blue),
3/16"
580Tubing connec-
Narrow inlet tubing is available for S1-S7. Refer to Section 9.3 Tubing and connectors, on
page 420 for more information.
46 ÄKTA pure User Manual 29119969 AB
2.4.5 Mixer valve
Function of Mixer valve
Mixer valve (V9-M or V9H-M) allows the user to bypass the mixer. It is intended to be used when the
System pump is used for sample application or when a sample is re-injected.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.5 Mixer valve
Note:
Mixer valve (V9-M or V9H-M) cannot be used together with Sample pump S9 or Sample pump
S9H.
Location and illustration of Mixer valve
The illustration below shows the recommended location, together with a detailed view of the Mixer
valve.
Ports and flow paths of the mixer valve
The illustrationand tables belowdescribe the ports of and different flowpathsthrough the Mixervalve.
The Mixervalve has two availableflow paths; By-pass, and Mixer(default).If the Mixer valve isinstalled
in the recommendedlocation before the Mixer, By-pass allows the flow to bypassthe Mixer, and Mixer
directs the flow to the Mixer.
ÄKTA pure User Manual 29119969 AB 47
Primary flow path
Alternative flow path
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.5 Mixer valve
DescriptionPort
In
Out
Connect tubing
The table below shows recommended tubing and connectors.
label
3-1
Pressure monitor R9
to
Mixer valve V9-Mport
In
Port in which the flow enters the valve. Should be connected to the System pressure
monitor outlet.
Port from which the flow leaves the Mixervalve and bypassesthe Mixer. Connect to the
injection valve SaP port.
Port for advanced configurations including re-injection.Re-inj
Port from which the flow leaves the valve and is directed to the Mixer.Mixer
ConnectorTubingConnectionTubing
Tubing
length
PEEK, o.d. 1/16",
i.d. 0.75 mm
ÄKTA pure 150ÄKTA pure 25
PEEK, o.d. 1/16",
i.d. 1.00 mm
1/16"
(mm)
160Fingertight connector,
3-2
Mixer valve V9-Mport
Mixer
PEEK, o.d. 1/16",
i.d. 0.75 mm
PEEK, o.d. 1/16",
i.d. 1.00 mm
1/16"
330Fingertight connector,
to Mixer M9
3-3
Mixer valve V9-Mport
Out to
PEEK, o.d. 1/16",
i.d. 0.75 mm
PEEK, o.d. 1/16",
i.d. 1.00 mm
1/16"
260Fingertight connector,
Injection valve V9-Inj
port SaP
48 ÄKTA pure User Manual 29119969 AB
Flow paths through Mixer valve
Primary flow path
Alternative flow path
The Mixer valve (V9-M, V9H-M) has two available flow paths; By-pass, and Mixer. If the Mixer valve is
installed in the recommended location before the Mixer, By-pass allows the flow to bypass the Mixer,
and Mixer directs the flow to the Mixer.
The illustration below shows the different flow paths through the Mixer valve V9-M.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.5 Mixer valve
DescriptionFlow path
The flow from the system pumps bypasses the mixer.By-pass
The flowfrom the systempumps is directed to the Mixer. Mixer is thedefaultflow path.Mixer
ÄKTA pure User Manual 29119969 AB 49
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.6 Injection valve
2.4.6 Injection valve
Function of the Injection valve
The Injection valve is used to direct sample onto the column. The valve enables usage of a number of
different sample application techniques.
The injection valve is labeled V9-Inj for ÄKTA pure 25 and V9H-Inj for ÄKTA pure 150.
A sample loop or a Superloop can be connected to the injection valve and filled either automatically,
using aSample Pump or System pumpA, or manually, usinga syringe. Thesample can also be injected
directly onto the column using a Sample pump, or System pump A together with the mixer valve.
For instructions on how to connect and use loops, see Section 5.7 Sample application, on page 180.
Location and illustration of the Injection valve
The illustration below shows the location, together with a detailed view of the Injection valve, in this
exampel labeled V9-Inj.
Ports and flow paths of the Injection valve
The illustration and tables below describe the ports of and different flow paths through the Injection
valve.
The Injection valve can be set to different positions that give different flow paths through the valve.
50 ÄKTA pure User Manual 29119969 AB
Primary flow path
Flow path for manual load
Closed flow path
Alternative flow path
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.6 Injection valve
ÄKTA pure User Manual 29119969 AB 51
DescriptionPort
Inlet fromSaP
•
sample pump, or
•
system pump via the Mixer valve Out port.
Inlet from the System pumps via the MixerSyP
Syringe connectionSyr
Outlet to one of the Column valves or to the column.Col
Port for connection of a loop. Used to fill the loop.LoopF
Port for connection of a loop. Used to empty the loop into the flow path.LoopE
Loop and System pump wasteW1
Sample flow wasteW2
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.6 Injection valve
DescriptionFlow path
Manual load - Default position of the valve
Inject
System pump waste
Direct inject
Sample pump load
The systemflow is directed ontothe column orcolumn valve. Sample
can be manually injected into the loop.
Excess sample leaves the valve through waste port W1.
The system flow is directed through the loop and onto the column
or column valve. If a mixer valve or the sample pump is used, the
flow entering the SaP port is directed to waste port W2.
The system flow is directed to waste port W1. If a mixer valve or the
sample pump is used, the flow entering the SaP port is directed to
the column or the column valve.
The flow entering the SaP port is directed to the column or the column valve. This position is used with:
•
the sample pump, or
•
a mixer valve and System pump A, bypassing the mixer.
Flow entering the SyP port is directed to waste port W1.
The flow entering the SaP port is directed to the loop. This position
is used with:
•
the sample pump, or
•
a mixer valve and System pump A, bypassing the mixer.
Sample pump waste
Note: •
In order to avoid sample carry-over when switching techniques for loading samples, wash
the injection valve with buffer between the loading of two different samples. For example,
when switching from loading sample onto the loop to loading sample directly onto the
column with the valve in Direct inject position.
•
Make sure that the SaP port is plugged with a stop plug if neither the sample pump nor
the mixer is used.
Excess sample leaves the valve through waste port W1. The flow
entering the SyP port is directed to the column or the column valve.
The flow entering the SaP port is directed to waste port W2. This
position is used with:
•
the sample pump, or
•
a mixer valve and System pump A.
The flowentering the SyPport is directedto the columnvia the loop.
52 ÄKTA pure User Manual 29119969 AB
2.4.7 Loop valve
Function of the Loop valve
The Loop valve allows the user to connect several loops simultaneously to the instrument. It can for
example be used for storing intermediate fractions in multi-step purifications, for storing samples to
be used in scouting runs, or for storing eluents needed in low volumes. The valve also has a built-in
bypass function that enables bypassing all loops. The Loop valve is labeled V9-L or V9H-L.
Location and illustration of Loop valve
The illustration below shows the recommended location, together with a detailed view of Loop valve
V9-L.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.7 Loop valve
Ports and flow paths of the Loop valve
The illustration and tables below describe the ports and different flow paths through the Loop valve.
In the Position 4 example, the
loop is connected to loop position 4 and the loop is being
emptied.
ÄKTA pure User Manual 29119969 AB 53
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.7 Loop valve
DescriptionPort
Port connected to the LoopF port of the Injection valve.F
Ports for connection to loop 1.1F and 1E
Ports for connection to loop 2.2F and 2E
Ports for connection to loop 3.3F and 3E
Ports for connection to loop 4.4F and 4E
Ports for connection to loop 5.5F and 5E
Port connected to the LoopE port of the Injection valve.E
Note:
Connect tubing
The table below shows recommended tubing and connectors.
label
L1
L2
Injection valve position LoopF to
Loop valve position F
Injection valve position LoopE to
Loop valve position E
Ports denoted by the letter F are used for filling the loop and ports denoted by the letter E are
used for emptying the loop.
DescriptionFlow path
The flow direction depends on the Injection valve position.Position 1-5
The flow bypasses the loop(s). By-pass is the default flow path.By-pass
ConnectorTubingConnectionTubing
Tubing
length
PEEK,
o.d. 1/16",
i.d. 0.50 mm
PEEK,
o.d. 1/16",
i.d. 0.50 mm
ÄKTA pure 150ÄKTA pure 25
PEEK,
o.d. 1/16",
i.d. 0.75 mm
PEEK,
o.d. 1/16",
i.d. 0.75 mm
1/16"
1/16"
(mm)
160Fingertight connector,
160Fingertight connector,
54 ÄKTA pure User Manual 29119969 AB
Connect a Loop valve
The Loop valve is connected to the Injection valve instead of a loop, as described below.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.7 Loop valve
ActionStep
Connect port E on the Loop valve to port LoopE on the Injection valve.1
Connect port F on the Loop valve to port LoopF on the Injection valve.2
3
Note:
Connect one or many loops to the Loop valve. See Section 5.7 Sample application, on
page 180.
Note:
Always use the the first positions of the valve for the connected loops (e.g., if three loops
will be used, use port 1F-3F and the corresponding ports 1E-3E) to avoid cross-contamination.
It is possible to place the Loop valve in other positions in the flow path than the one described
above. However, the volume used for washes will then be incorrect, just as the system configuration shown in the process picture.
ÄKTA pure User Manual 29119969 AB 55
1 2
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.8 Column valves
2.4.8 Column valves
Function of the Column valves
The Column valvesare used to connect columnsto the system, and to directthe flow onto the column.
The Column valves available for ÄKTA pure and their functions are described in the table below.
FunctionLabel in
ÄKTA pure 150ÄKTA pure 25
V9H-CV9-C
V9H-CsV9-Cs
The inletand outlet ports of Columnvalves V9-C and V9H-Chave built inpressure sensors thatmeasures
the actual pressure over the column. For further information on the pressure sensors, see Function of
pressure monitors integrated in Column valves V9-C or V9H-C, on page 69.
Column valves V9-Cs and V9H-Cs lack built-in pressure sensors. See Section 5.5 Connect a column, on
page 173 for information on how to set pressure alarms that protects the column when no Column
valve V9-C or V9H-C is installed.
Location and illustration of Column valves
The illustration below shows the recommended location of the Column valves and a detailed view of
the valves. In this example the valves are labeled V9-Cs and V9-C.
Connects columnsto the system andallows the user
to choosecolumn, flow directionthrough the column,
or to bypass the columns.
Up to five columns can be connected to the valve.
Connects columnsto the system andallows the user
to choose flow direction through the column, or to
bypass the column.
One column can be connected to the valve.
FunctionPart
Column valve V9-Cs or V9H-Cs (no integrated pressure sensors)1
56 ÄKTA pure User Manual 29119969 AB
By-pass
By-pass
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.8 Column valves
FunctionPart
Column valve V9-C or V9H-C (integrated pressure sensors)2
Note:
Maximum one column valve can be installed in ÄKTA pure at any given time.
Ports and flow paths of
Column valves V9-C and V9H-C
The illustration and tables below describethe different ports ofand flow paths through Column valves
V9-C and V9H-C. In the example below the column is connected to column position 4 and the valve is
labeled V9-C.
DescriptionPort
Down flow
Inlet from Injection valve via a built-in pressure monitor.In
Ports for connection to the top of columns.1A-5A
Ports for connection to the bottom of columns.1B-5B
Outlet to UV monitor via a built-in pressure monitor.Out
DescriptionFlow path
The flow bypasses the column(s). By-pass is the default flow path.By-pass
The flow direction is from the top of the column to the bottom of the column.
Down flow is the default flow direction.
The flow direction is from the bottom of the column to the top of the column.Up flow
ÄKTA pure User Manual 29119969 AB 57
1A
1B
1A
1B
1A
1B
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.8 Column valves
Ports and flow paths of Column valves V9-Cs and
V9H-Cs
The illustration and tables below describe the different ports and flow paths of Column valve V9-Cs.
DescriptionPort
path
Down
flow
Connect tubing
The table below shows recommended tubing and connectors.
Inlet from Injection valve.In
Port for connection to the top of a column.1A
Port for connection to the bottom of a column.1B
Outlet to UV monitor.Out
DescriptionFlow
The flow bypasses the column. By-pass is the default flow path.By-pass
The flowdirection is from the top of thecolumnto the bottomofthe column. Downflow
is the default flow direction.
The flow direction is from the bottom of the column to the top of the column.Up flow
ConnectorTubingConnectionTubing
label
ÄKTA pure 150ÄKTA pure 25
5
Injection valve to
Column valve
6
Column valve to
UV monitor
58 ÄKTA pure User Manual 29119969 AB
PEEK, o.d. 1/16",
i.d. 0.50 mm
PEEK, o.d. 1/16",
i.d. 0.50 mm
PEEK, o.d. 1/16",
i.d. 0.75 mm
PEEK, o.d. 1/16",
i.d. 0.75 mm
1/16"
1/16"
Tubing
length
(mm)
100Fingertight connector,
160Fingertight connector,
Follow the instructions below to connect tubing to the column valves.
ActionStep
If no Column valve is installed, remove the Union F/F between tubing 5 and tubing 61
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.8 Column valves
2
Note:
Connect tubing between Injection valve, Column valve and UV monitor
according to the table above.
The built-in pressure sensors for Column valve V9-C and V9H-C have to be re-calibrated after
installation. See Calibrate the monitors, on page 305.
ÄKTA pure User Manual 29119969 AB 59
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.9 Versatile valve
2.4.9 Versatile valve
Function of the Versatile valve
The Versatile valve is a 4-port, 4-position valve, which can be used to add extra features to the flow
path. For example, the valve can be used to connect external equipment to the flow path during parts
of a run. The versatile valve can be installed in any position. The versatile valve is labeled V9-V or V9H-
V.
It ispossible to installup to fourversatilevalves simultaneously in ÄKTA pure.The configuration isdefined
by the module's Node ID.
Ports and flow paths of the Versatile valve
The illustration and table below describe the different ports of and flow paths through the Versatile
valve. The valve has four ports (1-4).
The Versatile valve has four available sets of flow paths; two where a single flow channel is used and
two where the flow can be directed through two different channels simultaneously.
DescriptionFlow
path
A single flow channel where the flow is directed between port 1 and port 3.1-3
A single flow channel where the flow is directed between port 2 and port 4.2-4
1-4 and
2-3
1-2 and
3-4
Two simultaneously used flow channels where the flow is directed between port 1 and
port 4 and between port 2 and port 3.
Two simultaneously used flow channels where the flow is directed between port 1 and
port 2 and between port 3 and port 4.
60 ÄKTA pure User Manual 29119969 AB
2.4.10 pH valve
1
2
3
4
Function of the pH valve
The pH valve is usedto direct the flow to a pH electrode when inline monitoring of pH is desired during
a run. The pH valve is labeled V9-pH for ÄKTA pure 25 and V9H-pH for ÄKTA pure 150.
The pH valve has an integrated flow cell in which the pH electrode can be installed.
It is recommended to connect a Flow restrictor to the pH valve. The flow restrictor is used to generate
a back pressure high enough to prevent the formation of air bubbles in the UV flow cell.
The valve directs the flow to the pH electrode and to the flow restrictor, or bypasses one or both.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.10 pH valve
Note:
The Flow restrictor is normally included in the flow path after the Conductivity monitor. When
installing the pH valve on ÄKTA pure, the flow restrictor should be moved from its normal position on the conductivity monitor to the pH valve.
Location and illustration of the pH valve
The illustration below shows the recommended location, together with a detailed view of
the pH valve.
DescriptionPart
pH valve1
pH electrode2
pH flow cell3
Flow restrictor4
ÄKTA pure User Manual 29119969 AB 61
By-pass
Primary flow path
Flow path for calibrations
Closed flow path
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.10 pH valve
Ports and flow paths of the pH valve
The illustration and table below describe the different ports of and flow paths through the pH valve,
in this example labeled V9-pH.
DescriptionPort
From Conductivity monitorIn
To Flow restrictorToR
From Flow restrictorFrR
To Outlet valveOut
Calibration portCal
To WasteW3
62 ÄKTA pure User Manual 29119969 AB
and pH
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.10 pH valve
DescriptionFlow path
Both pH electrode and Flow restrictor are bypassed.By-pass
Flow restrictor is in use and pH electrode is bypassed.Restrictor
Both pH electrode and Flow restrictor are in use.Restrictor
pH electrode is in use and Flow restrictor is bypassed.pH
pH monitor
Calibration
Flow path used when calibrating the pH monitor and when filling the pH flow cell
with storage solution. The Cal port is used to inject solution into the flow cell using
a syringe.
Excess solution leaves the valve through port W3.
The pHmonitor continuously measuresthe pH of thebufferand eluted proteinswhen the pH electrode
is inline. A pH electrode can be installed in the pH flow cell. For instruction on how to install the pH
electrode, see Section 7.5.2 Replace the pH electrode, on page 275. The pH electrode should not be exposed to more than 0.5 MPa during a normal run, but can withstand pressure spikes of 0.8 MPa. It is
therefore important to place it after the column.
The illustration below shows the location of the pH flow cell and a pH electrode installed in the pH
valve.
DescriptionPart
pH flow cell1
pH electrode2
ÄKTA pure User Manual 29119969 AB 63
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.10 pH valve
Connect tubing
The table below shows recommended tubing.
label
8pH
1R
2R
9pH
Conductivity monitor
to port In
Flow restrictor to
port ToR
Flow restrictor to
port FrR
Port Out to
Outlet valve
Note:
The dimensions of the PEEK tubing depends on which tubing kit that is used. See Section 9.3
Tubing and connectors, on page 420 for more information.
PEEK, o.d. 1/16",
i.d. 0.50 mm
PEEK, o.d. 1/16",
i.d. 0.50 mm
PEEK, o.d. 1/16",
i.d. 0.50 mm
PEEK, o.d. 1/16",
i.d. 0.50 mm
ÄKTA pure 150ÄKTA pure 25
PEEK, o.d. 1/16",
i.d. 0.75 mm
PEEK, o.d. 1/16",
i.d. 0.75 mm
PEEK, o.d. 1/16",
i.d. 0.75 mm
PEEK, o.d. 1/16",
i.d. 0.75 mm
ETFE I.D. 1 mmETFE I.D. 1 mmPort W3 to WasteW3
ConnectorTubingConnectionTubing
1/16"
1/16"
1/16"
1/16"
1/16"
Tubing
length
(mm)
180Fingertight connector,
80Fingertight connector,
80Fingertight connector,
160Fingertight connector,
N/AN/AN/AN/AtoCalibrationport CalN/A
1500Fingertight connector,
64 ÄKTA pure User Manual 29119969 AB
2.4.11 Outlet valves
Function of the outlet valves
The outlet valve is used to direct the flow to the fraction collector, to an outlet port, or to waste. The
table below shows the labeling of the outlet valves for ÄKTA pure 25 and ÄKTA pure 150.
ÄKTA pure 150ÄKTA pure 25
Location and illustration of outlet valves
The illustration below shows the recommended location, together with a detailed view of the outlet
valves. In this example the valves are labeled V9-Os and V9-O.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.11 Outlet valves
DescriptionLabel in
10 outlet portsV9H-OV9-O
1 outlet portsV9H-OsV9-Os
Ports of Outlet valves V9-O and V9H-O
The illustration below shows the ports of Outlet valve V9-O and V9H-O, in this example labeled V9-O.
DescriptionPort
Inlet portIn
ÄKTA pure User Manual 29119969 AB 65
In
Out1Frac
W
V9-Os
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.11 Outlet valves
Out10
DescriptionPort
Outlet ports 1 - 10Out1 -
Frac
Port to Fraction collector
Note:
If a secondary Fraction collector F9-R is used it should be connected to port Out10.
Waste portW
Ports of Outlet valves V9-Os and V9H-Os
The illustration below shows the ports of Outlet valve V9-Os.
DescriptionPort
Inlet portIn
Outlet portOut1
Frac
Port to fraction collector
Note:
If a secondary FractioncollectorF9-Ris used it should be connected to port
Out1.
Waste portW
66 ÄKTA pure User Manual 29119969 AB
Connect tubing
The table below shows recommended tubing and connectors.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.11 Outlet valves
label
9
9pH
Out1 Out10
Flow restrictor to
Outlet valve
pH valve to
Outlet valve
Outlets from the
outlet valves
Note:
If ÄKTA pure previously has been configured without an Outlet valve, remove the Union F/F
between tubing 9 and tubing W before installation of the Outlet valve. Then connect tubing 9
to the In port on the Outlet valve and the waste tubing W to the W port on the Outlet valve.
PEEK, o.d. 1/16",
i.d. 0.50 mm
PEEK, o.d. 1/16",
i.d. 0.50 mm
ETFE, o.d. 1/8",
i.d. 1 mm
ÄKTA pure 150ÄKTA pure 25
PEEK, o.d. 1/16",
i.d. 0.75 mm
PEEK, o.d. 1/16",
i.d. 0.75 mm
ETFE, o.d. 1/8",
i.d. 1 mm
ConnectorTubingConnectionTubing
1/16"
1/16"
1/16"
Tubing
length
(mm)
135Fingertight connector,
160Fingertight connector,
1500Fingertight connector,
ÄKTA pure User Manual 29119969 AB 67
1
2
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.12 Pressure monitors
2.4.12 Pressure monitors
Introduction
This section describes the location and function of the pressure monitors.
Up to four pressure monitors are included in ÄKTA pure.
Function of the system pump and the sample
pump pressure monitors
Up to four pressure monitors are included in ÄKTA pure. One pressure monitor is always connected
to the system pumps and one pressure monitor is part of the Sample pump (S9 or S9H). The monitors
of the pumps are both labelled R9.
The System pump pressure monitor measures the pressure after the system pumps, the system
pressure. The pressure monitor of the sample pump measures the pressure after the sample pump,
the sample pressure. To provide a reading of the highest pressure in the system there are connections
to all pump cylinders.
The pressure monitor also contains pump flow restrictors in orderto stop siphoning effects.Siphoning
effects can occur if tubing in the system is disconnected while the inlet tubing is immersed in buffers
that are placed on a higher level than the pump.
Location and illustration of System pressure
monitor
The illustration below shows the location, together with a detailed view of System pressure monitor.
DescriptionPart
Pressure monitor1
Pump flow restrictors2
68 ÄKTA pure User Manual 29119969 AB
Function of pressure monitors integrated in
Column valves V9-C or V9H-C
If Column valve C, labeled V9-C or V9H-C, is used, the two pressure monitors integrated in
the valve can be used to monitor the column pressure.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.12 Pressure monitors
Note:
Column valve V9-Cs and V9H-Cs does not contain pressure monitors.
The pressure sensors integrated in the inlet and outlet ports of Column
valve C measure the pressure before the column, Pre column pressure (A),
and the pressure after the column, Post column pressure (B).
The Delta columnpressure (C),which is thepressure dropover the column,
is calculated as the difference between the Pre column pressure and the
Post column pressure.
Pressure alarmscan be setforboth the Precolumn pressureand the Delta
column pressure. Pressure control of the flow may be based on either the
Pre column pressure or the Delta column pressure.
ÄKTA pure User Manual 29119969 AB 69
1 2
34
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.13 UV monitors
2.4.13 UV monitors
Introduction
This section describes the design and function of UV monitors U9-M and U9-L.
The modules include a monitor unit and a detector with a UV flow cell.
Function of UV monitor U9-M
UV monitor U9-M measures the UV absorbance at a wavelength range of 190 to 700 nm.
A flip-mode enables measuring of UV/Vis absorbance at three wavelengths simultaneously during a
run. The second and third wavelength can be turned off or on in method phase properties, by manual
instructions or in system settings.
Note:
Note:
Installation of UV monitor U9-M should only be performed by GE Service personnel.
The resolution is decreased when more than one wavelength is used simultaneously due to
lower sampling frequency per wavelength. Do not use more wavelengths than necessary.
Location and illustration of UV monitor U9-M
The illustration below shows the location of UV monitor U9-M, together with a detailed view of the
monitor unit and detector. The monitor unit is labelled U9-M and the detector U9-D.
Note:
When UV monitor U9-M is used, the entire Multi-module panel shown in the illustration is replaced by U9-M.
DescriptionPart
Inlet1
2
70 ÄKTA pure User Manual 29119969 AB
UV flow cell. Three different path lengths are available:
0.5 mm, 2 mm (default) and 10 mm
UV detector3
Outlet4
Function of UV monitor U9-L
1 2 3
45
The UV monitor U9-L measures the UV absorbance at the fixed wavelength of 280 nm.
It is not possible to vary the wavelength,or turn on or off the U9-L monitor. This is therefore not shown
in the Phase Properties pane in Method Editor.
Location and illustration of UV monitor U9-L
The illustration below shows the recommended location of UV monitor U9-L, together with a detailed
view ofthe monitor anddetector. TheUV monitor U9-Lrequires thatthe Multi-module panelis installed,
see Example of a typical configuration of the wet side, on page 16. The UV monitorcan also be installed
in an Extension box, see Extension box, on page 86.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.13 UV monitors
DescriptionPart
Inlet1
UV monitor U9-L2
3
UV flow cell. Two different path lengths are available:
2 mm (default) and 5 mm
Outlet4
UV detector5
Using two UV monitors
It is possible to use two UV monitors in ÄKTA pure, in two combinations. The configuration is defined
by the module's Node ID.
•
UV monitor U9-M together with UV monitor U9-L, 2nd
•
UV monitor U9-L together with UV monitor U9-L, 2nd
ÄKTA pure User Manual 29119969 AB 71
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.13 UV monitors
Note:
Note:
UV monitor U9-L, 2nd canbe located anywhere in the flowpath and is therefore shown in the Process
Picture in UNICORN as a component without a fixed place. This means that it is possible to place U9-
L, 2nd before the other UV monitor in the flow path.
Note:
Connect tubing
The table below shows the tubing and connectors to be used with UV monitor U9-L.
Note:
label
When using two UV monitors, the signal from the first UV monitor is by default used for peak
fractionation. This can be changed by editing the text instruction Fraction Collection:Peak
fractionation parameters:Signal source and choosing UV 2nd as Signal source.
When using two UV monitors withdifferent cell lengths to increase the UV absorption dynamic
range, the U9-L signal comes from the real cell length and has to be calibrated for exact calculations. The U9-M signal is automatically calibrated to nominal cell length.
If U9-L 2nd is placed on the high pressure side of the column, pressure limits have to be considered. See UV monitor options, on page 416
If a second UV monitor is used, the tubing for this has to be cut manually.
ConnectorTubingConnectionTubing
Tubing
length
ÄKTA pure 150ÄKTA pure 25
(mm)
6
Tubing to
UV monitor U9-L
7
Tubing from
UV monitor U9-L
Note:
To perform a run with the flow in reverse direction through UV monitor U9-L, a longer tubing
PEEK, o.d. 1/16",
i.d. 0.50 mm
PEEK, o.d. 1/16",
i.d. 0.50 mm
PEEK, o.d. 1/16",
i.d. 0.75 mm
PEEK, o.d. 1/16",
i.d. 0.75 mm
1/16"
1/16"
160Fingertight connector,
170Fingertight connector,
7 is needed. Replace the 170 mm tubing from the UV monitor with tubing that is 210 mm long
and adjust the delay volume accordingly. For example, changing from 170 mm to 210 mm for
0.5 mm i.d. tubing increases the delay volume with 8 µl.
72 ÄKTA pure User Manual 29119969 AB
2.4.14 Conductivity monitor
1
2
3
4
Function of the Conductivity monitor
The Conductivity monitor continuously measures the conductivity of buffers and eluted proteins. The
monitor is labelled C9.
The Conductivity flow cell has two electrodes positioned in the flow path of the cell. An alternating
voltage is applied between the electrodes and the resulting current is measured and used to calculate
the conductivity of the eluent.
The conductivity is automatically calculated by multiplying the measured conductance by the cell
constant of the flow cell. The cell constant is factory-calibrated on delivery but can be re-calibrated if
needed, see Section 7.7.3 Calibrate the Conductivity monitor, on page 308.
As variation in temperature influences conductivity readings, the conductivity flow cell is fitted with a
temperature sensorthat measures thetemperature oftheeluent. A temperaturecompensation factor
is used to report the conductivity in relation to a set reference temperature.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.14 Conductivity monitor
Location and illustration of the
Conductivity monitor
The illustration below shows the recommended location of Conductivity monitor C9, together with a
detailed view of the monitor.
DescriptionPart
Inlet1
Conductivity flow cell2
Conductivity monitor3
Outlet4
ÄKTA pure User Manual 29119969 AB 73
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.14 Conductivity monitor
Connect tubing
The table below shows recommended tubing and connectors.
label
7
8
UV monitor U9-L to
Conductivity monitor C9
Conductivity monitorC9 to
Flow restrictor
PEEK, o.d. 1/16",
i.d. 0.50 mm
PEEK, o.d. 1/16",
i.d. 0.50 mm
ÄKTA pure 150ÄKTA pure 25
PEEK, o.d. 1/16",
i.d. 0.75 mm
PEEK, o.d. 1/16",
i.d. 0.75 mm
ConnectorTubingConnectionTubing
1/16"
1/16"
Tubing
length
(mm)
170Fingertightconnector,
95Fingertight connector,
74 ÄKTA pure User Manual 29119969 AB
2.4.15 Flow restrictor
1
4 3
2
Function of Flow restrictor FR-902
The Flow restrictor is included in the flow path to generate a steady back pressure of approximately
0.2 MPa, to prevent formation of air bubbles in the UV flow cell.
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.15 Flow restrictor
Note:
Do not remove the flow restrictor to lower the pressure in the system. There is a risk that air
bubbles cause large disturbances in the UV flow cell. Use the automatic pressure control
function to avoid pressure alarms, see Recommended pressurecontrol parameters, on page502
Location and illustration of Flow restrictor FR-902
The Flow restrictor is normally included in theflow path after the Conductivity monitor. The Conductivity monitor is equipped with a special holder for the Flow restrictor.
The illustration below shows Flow restrictor FR-902 fitted on the Conductivity monitor.
FunctionPart
Flow restrictor1
Holder2
Inlet3
If ÄKTA pure is fitted with pH valve V9-pH or V9H-pH the Flow restrictor has to be moved from the
Conductivity monitor to the pH valve.
ÄKTA pure User Manual 29119969 AB 75
Outlet4
1
2
3
2 The ÄKTA pure instrument
2.4 Instrument modules
2.4.15 Flow restrictor
The illustration below shows Flow restrictor FR-902 fitted on the pH valve.
FunctionPart
Flow restrictor1
Flow restrictor inlet connection from pH valve ToR port2
Flow restrictor outlet connection to pH valve FrR port3
76 ÄKTA pure User Manual 29119969 AB
2.5 Installation of internal modules
A B
Introduction
Optional modules and valves are easy to install in the instrument. The existing module or Module
Panel isremoved with aTorx T20 screwdriverand the cableis disconnected. The cableis then connected
to the optionalmodule,which is subsequentlyinserted intothe instrument. The newly installedmodule
is then added to the System properties in UNICORN.
Node ID
All of the available optional modules are preconfigured to give the desired function. However, the
function of a module or valve can be changed by changing its Node ID. Node ID is a unit number designation that is used by the instrument to distinguish between several units of the same type.
In a troubleshooting situation it may be useful to check a valve's or module's Node ID. Refer to Sec-
tion 9.16 Node IDs, on page 503 for a list of Node IDs for valves and modules.
2 The ÄKTA pure instrument
2.5 Installation of internal modules
Note:
The Node ID is set by positioning the arrows of the one or two rotating switches at the back of the
valve. Use a screwdriver to set the arrows of the switches to the desired number.
The function of a valve or module is defined by its Node ID, not by its physical position.
Hardware installation of a module
The instruction below describes how to install a module in the instrument.
Note:
The illustrations show the principle how to install an optional module. The position of the
module on the instrument and the used type of module will depend on the module being installed.
The illustration shows an example of a valve module with
two rotating switches.
•
The first rotating switch, labeled A, sets the tens
•
The second switch, labeled B, sets the units.
CAUTION
Disconnect power. Always switch off power to the ÄKTA pure instrument before
replacing any of itscomponents, unless statedotherwise inthe user documentation.
ÄKTA pure User Manual 29119969 AB 77
2 The ÄKTA pure instrument
2.5 Installation of internal modules
ActionStep
Disconnect power from the instrument by switching off the instrument power switch.1
2
Loosen the connectors and remove the tubing from the existing module.
Note:
This step does not apply for a Module Panel.
Loosen the module with a Torx T20 screwdriver.3
Remove the module.4
Disconnect the cable and secure it in the slit.5
78 ÄKTA pure User Manual 29119969 AB
2 The ÄKTA pure instrument
2.5 Installation of internal modules
ActionStep
Connect the cable to the module to be installed.6
Insert the module.7
Fasten it with a Torx T20 screwdriver.8
Note:
A warning message is displayed at start up if a module has been installed in the instrument
but not added to the current system configuration in UNICORN.
Install internal modules in the Extension box
The Extension box can be used to install extra modules on the ÄKTA pure instrument. See Extension
box, on page 86 for more information.
ÄKTA pure User Manual 29119969 AB 79
1 2
1 2
3 4 3
2 The ÄKTA pure instrument
2.6 Accessories
2.6 Accessories
Introduction
This sectiondescribes the holders andother available accessories.Theseare used toattach and organize
columns, tubing and bottles to the ÄKTA pure instrument. The holders are attached to the instrument
using the holder rails on the left side and the front of the instrument.
Rail extension
The Rail extension rod can be used to attach accessories, eg., column holders or a Multi-purpose
holder. The rod has extra rails on both sides. Push the button of the rod to attach it to a holder rail.
The illustration below shows the Rail extension rod.
Multi-purpose holder
The Multi-purpose holder can be used to attach accessories, eg., a Loop holder or a cassette. Attach
the holder to a holder rail.
The illustration below shows the Multi-purpose holder.
FunctionPart
Button1
Extension rod2
FunctionPart
Attachment point for accessories1
Snap-in to holder rails2
80 ÄKTA pure User Manual 29119969 AB
3
Attachment points for tubing holders
Tab4
Loop holder
1
2
1
2
3
4
2 The ÄKTA pure instrument
2.6 Accessories
The Loop holder can be used to attach up to five 10 ml sample loops. Use two Multi-purpose holders
to attach the holder to a holder rail.
The illustration below shows the Loop holder.
FunctionPart
Upper attachment to multi-purpose holder1
Lower attachment to multi-purpose holder2
Column holder
The Columnholder has one positionfor medium sizedcolumns and one positionfor small sizedcolumns.
The Column holder can also be used for bottles. Use two holders to attach long columns.
The illustration below shows the Column holder.
DescriptionPart
ÄKTA pure User Manual 29119969 AB 81
1
Position for a medium sized column
or bottle
Position for a small sized column2
Tab3
Snap-in to holder rails4
1
2
1
2 3
2 The ÄKTA pure instrument
2.6 Accessories
Column clamp
The columnclamp can be used to attachsmall sized columns.Use two clamps to attach longcolumns.
The illustration below shows the Column clamp.
DescriptionPart
Position for a column1
Inner end tabs2
Column holder rod
The Columnholder rod can be used to attachseveralHiTrap™ columns. Theholder has threaded ports
for HiTrap columns and tubing connectors. Push the button of the holder to attach the holder to a
holder rail.
The illustration below shows the Column holder rod.
DescriptionPart
Snap-in to holder rails1
Button2
Column holder rod3
82 ÄKTA pure User Manual 29119969 AB
Flexible column holder
2
3
4
5
7
6
9
1
8
1
2 3
The Flexible column holder can be used to attach, for example, HiScreen™ columns.
The illustration below shows the Flexible column holder.
2 The ÄKTA pure instrument
2.6 Accessories
FunctionPart
Lower tubing1
Lower part2
Snap-in-strips3
Attachment part4
Upper tubing5
Lever6
Upper part7
Upper connector8
Lower connector9
Tubing holder spool
The Tubing holder spool is used to hold and arrange tubing.
The illustration below shows the Tubing holder spool.
DescriptionPart
Positions for tubing1
ÄKTA pure User Manual 29119969 AB 83
Tab2
Snap-in to holder rails3
1
2
3
1
2
1
2
3
2 The ÄKTA pure instrument
2.6 Accessories
Tubing holder comb
The Tubing holder comb is used to hold and arrange tubing.
The illustration below shows the Tubing holder comb.
Bottle holder
The Bottle holder is used for holding bottles. For example, the Bottle holder can be attached to the
holder rails to hold a sample bottle.
DescriptionPart
Positions for tubing1
Tab2
Snap-in to holder rails3
The illustration below shows the Bottle holder.
Adapter for air sensor
The adapter for air sensor is used to hold an optional air sensor.
The air sensor with adapter is connected to the Bottle holder, see illustrations below.
DescriptionPart
Position for bottle1
Snap-in to holder rails2
DescriptionPart
Air sensor1
Air sensor adapter2
Bottle holder3
84 ÄKTA pure User Manual 29119969 AB
Module Panel
Description
All positions in ÄKTA pure must be occupied. Positions not used for core or optional modules must be
fitted with a Module Panel.
Module Panels are installed in the same way as the other optional modules and the cable inside must
be connected to the Module Panel, see Hardware installation of a module, on page 77.
When an optional module is replaced by a Module panel, the removed module has to be deselected
in the software configuration, see Edit system properties, on page 128.
Illustration
The illustration shows the Module Panels installed in the example configuration used in this manual.
2 The ÄKTA pure instrument
2.6 Accessories
ÄKTA pure User Manual 29119969 AB 85
2 The ÄKTA pure instrument
2.6 Accessories
Extension box
Description
The Extensionbox can be used toinstallextra modules onthe ÄKTA pureinstrument outside the system
chassis when the positions on the chassis are filled. It is possible to install up to six Extension boxes
with extra modules when using ÄKTA pure. See the Extension box instruction for more information.
Location
The illustration below shows an Extension box with a module mounted on the side of ÄKTA pure.
The Extension box can be mounted in three ways.
•
On the side of ÄKTA pure.
•
Standing on top of or next to ÄKTA pure.
•
On a Rail extension rod (29-0113-52) on ÄKTA pure.
86 ÄKTA pure User Manual 29119969 AB
3 ÄKTA pure external modules
3 ÄKTA pure external modules
About this chapter
This chapter provides an overview of the external modules that can be connected to the ÄKTA pure
instrument. A brief description of how to connect external modules is also provided.
In this chapter
This chapter contains the following sections:
See pageSection
883.1 External air sensors
903.2 Fraction collector F9-C
1043.3 Fraction collector F9-R
1083.4 Sample pump S9 and S9H
1123.5 I/O-box E9
1223.6 Connection of external modules
ÄKTA pure User Manual 29119969 AB 87
3 ÄKTA pure external modules
3.1 External air sensors
3.1 External air sensors
Introduction
Up to four external air sensors can be added to ÄKTA pure, and there are two different versions to
choose from. They differ in internal diameter and optimal position on the instrument.
The airsensors can beattachedto the instrumentusing the railsand holders, seeAdapter forair sensor,
on page 84. No Module Panels need to be removed.
In additionto be used for preventing air fromenteringthe system, the externalairsensors canbeused
together with System pump A, or an Sample pump to load the entire sample volume.
Air sensor L9-1.5
L9-1.5 has a 1.5 mm inner diameter and is designed for i.d. 1.6 mm FEP, ÄKTA pure 25, and for i.d. 2.9
mm FEP tubing, ÄKTA pure 150, at the low pressure side before the pumps. It is installed in the flow
path before the system pumps or the samplepumpandis used to preventair entering thesubsequent
module.
Air sensor L9-1.2
L9-1.2 has a 1.2 mm inner diameterand is designedfor o.d. 1/16"tubing at the high pressureside after
the pumps. It is installed after the injection valve and is used to prevent air entering the column.
Note:
Tubing connections
Air sensor L9-1.5
between...
L9-1.5 and inlet
valves
FEP, o.d. 1/8", i.d.
1.6 mm
Note:
L9-1.2 can be used in the same fashion as L9-1.5 if O.D. 1/16" ETFE tubing is used between
the air sensor and the inlet valve.
ConnectorTubingConnection
ÄKTA pure 150ÄKTA pure 25ÄKTA pure 150ÄKTA pure 25
FEP, o.d. 3/16", i.d.
2.9 mm
When sample is loaded at high flow rate and the external air sensor is placed before the pump
that is used for loading the sample, it is necessary to use longer tubing to ensure that no air
reaches the pump.Use tubing with the minimum lengths givenbelow between the valve located
before the pump and the external air sensor. The length applies for maximum flow rate, but
shorter tubing can be used at lower flow rates.
Tubing connector,
5/16" + Ferrule
(yellow), 1/8"
5/16" + Ferrule
(blue), 3/16"
Tubing
length
(mm)
See noteTubing connector,
•
Sample pump S9: 40 cm.
•
Sample pump S9H: 20 cm.
88 ÄKTA pure User Manual 29119969 AB
Air sensor L9-1.2
3 ÄKTA pure external modules
3.1 External air sensors
between...
connected column
ConnectorTubingConnection
Tubing
length
(mm)
PEEK, o.d. 1/16"Injection valve and L9-1.2
100Fingertight connector,
1/16"
PEEK, o.d. 1/16"L9-1.2 and Column valve/the
100Fingertight connector,
1/16"
ÄKTA pure User Manual 29119969 AB 89
3 ÄKTA pure external modules
3.2 Fraction collector F9-C
3.2 Fraction collector F9-C
About this section
This section shows an overview of Fraction collector F9-C.
Technical details are found in the ÄKTA pure User manual.
In this section
This section contains the following subsections:
See pageSection
913.2.1 Function
933.2.2 Fraction collector F9-C illustrations
973.2.3 Cassettes, Cassette tray and racks
1033.2.4 Connect tubing to the ÄKTA pure instrument
90 ÄKTA pure User Manual 29119969 AB
3.2.1 Function
Introduction
Fraction collector F9-C can collect fractions in deep well plates, tubes of different sizes or bottles. Up
to six cassettes for deep well plates and tubes can be used. The cassettes can be used in any combination and are placed on the Cassette tray. A rack for 50 ml tubes and a rack for 250 ml bottles are
also available.
Scanner functions detect which types of trays, racks, cassettes and deep well plates that are used in
each run.
The Cassette tray or one of the racks is placed inside the fraction collector. A height exclusion bar ensures that the bottles, tubes and deep well plates are correctly positioned and cannot damage the
Dispenser head. The Tray catch and positioning discs on the floor of the fraction collector guide the
Cassette tray or the rack into the correct position.
Scanning of Cassettes
3 ÄKTA pure external modules
3.2 Fraction collector F9-C
3.2.1 Function
When thedoor of the fractioncollectoris closed automatic scanningisperformed. There are twotypes
of scanning procedures:
•
Full scan: Scanning of Cassette type codes to determine which types of Cassettes are used, and
scanning of rows and columns in deep well plates to identify which types of plates are used (24,
48, or 96 wells). Full scan is performed only when the system is in state Ready.
•
Quick scan: Scanning of Cassette type codes to determinewhich type of Cassettes are used. Quick
scan isperformed duringthe run toensure that correctCassettes are placedin the Fraction collector.
Fractionation modes to avoid spillage
Three fractionation modes are available, all of which avoid spillage between wells or tubes during
fractionation:
•
Accumulator: The accumulator is used to collect liquid during movement between wells, tubes or
bottles. The liquid is then dispensed in the next well or tube. Fractionation with accumulator can
be used at all flow rates.
•
DropSync: When using DropSyncthe sensors inthe Dispenser head detectwhen a drop isreleased
from the nozzle. The Dispenser head moves to the next well or tube just after a drop is released.
Fractionation with DropSync can be used at flow rates up to 2 ml/min. If the cassettes are placed
near the waste funnel the DropSync mode can be used at higher flow rates. Volatile solutions and
solutions with low surface tension may require a lower flow.
•
Automatic: The fraction collectoruses the Drop Sync mode for flow rates up to 2 ml/minand automatically switches to Accumulator mode for higher flow rates.
ÄKTA pure User Manual 29119969 AB 91
3 ÄKTA pure external modules
3.2 Fraction collector F9-C
3.2.1 Function
Fractionation arm positions
•
Home position: The homeposition is used when thefractioncollector is idle. The Fractionationarm
is positionedin the front ofthe interior ofthe fraction collector andtheDispenser head is positioned
over the waste funnel. This position is called Waste (Frac) in UNICORN.
•
Frac cleaning position: The Fraccleaningposition is used for convenient cleaningof the Dispenser
head. The Fractionation arm is positioned in the front of the interior of the fractionation collector
and the Dispenser head is moved to the center of the Fractionation arm.
92 ÄKTA pure User Manual 29119969 AB
3.2.2 Fraction collector F9-C illustrations
1 2 3 5 64
Introduction
This section provides illustrations of Fraction collector F9-C. The main features and components are
indicated.
Front view
The illustration below shows the main parts of the exterior of Fraction collector F9-C.
3 ÄKTA pure external modules
3.2 Fraction collector F9-C
3.2.2 Fraction collector F9-C illustrations
ÄKTA pure User Manual 29119969 AB 93
DescriptionPart
Fractionation indicator1
Symbol indicatingthat fractionation is ongoing. Donotopen the doorwhile the indicator
is lit.
Door2
Window3
Door handle4
Tubing connector for outlet valve tubing5
Vents6
1
1
23
3 ÄKTA pure external modules
3.2 Fraction collector F9-C
3.2.2 Fraction collector F9-C illustrations
Rear view
The illustration below shows the rear view of Fraction collector F9-C.
DescriptionPart
Vents1
UniNet-9 D-type connector (for communication and power supply)2
Waste tube3
94 ÄKTA pure User Manual 29119969 AB
Interior
1
2
3 4 5 6 7 8
9
10
11
12
1314
3 ÄKTA pure external modules
3.2 Fraction collector F9-C
3.2.2 Fraction collector F9-C illustrations
The illustration below shows the main parts of the interior of Fraction collector F9-C.
DescriptionPart
Fractionation arm guide rail1
Fractionation arm main rail2
Lamp3
Tubing guide4
Tubing connection5
Dispenser head6
Fractionation arm7
Tubing guide8
Height exclusion bar9
Waste funnel10
Waste tube11
Tray catch12
Waste groove, in case of overflow13
ÄKTA pure User Manual 29119969 AB 95
Tray guides14
1 2 3
4
56
3 ÄKTA pure external modules
3.2 Fraction collector F9-C
3.2.2 Fraction collector F9-C illustrations
Dispenser head
The illustration below shows the Dispenser head of Fraction collector F9-C.
DescriptionPart
Dispenser head1
Nozzle2
Dispenser head cover3
Accumulator (back part of Dispenser head)4
Drop sync sensor5
Type code reader6
96 ÄKTA pure User Manual 29119969 AB
3.2.3 Cassettes, Cassette tray and racks
Introduction
Fractions can be collected in deep well plates and in tubes of different sizes. A number of cassettes
and racks for different tubes and deep well plates are available. The cassettes are placed on a rack
with sixcassette positions. TheCassette type codesare scanned by the Cassette code readerto determine the type of Cassette.
Available cassettes, trays and racks
The following Cassettes and racks are available:
•
Cassette 3 ml tubes (for 40 tubes)
•
Cassette 5 ml tubes (for 40 tubes)
•
Cassette 8 ml tubes (for 24 tubes)
•
Cassette 15 ml tubes (for 15 tubes)
3 ÄKTA pure external modules
3.2 Fraction collector F9-C
3.2.3 Cassettes, Cassette tray and racks
•
Cassette 50 ml tubes (for 6 tubes)
•
Cassette for deep well plate (24, 48, 96 wells)
•
Cassette tray (for six cassettes)
•
Rack for 50 ml tubes (for 55 tubes)
•
Rack for 250 ml bottles (for 18 bottles)
For information on dimension requirements for tubes and deep well plates to be used in the fraction
collector, seeFraction collector tubes and bottles, on page 100 andDeep well plates, on page101 respectively.
Illustrations of Fraction collector F9-C tray and
racks
The illustrations belowshow the Cassettetray, the Rackfor 50 mltubesand the Rackfor 250 mlbottles.
The fronts of the tray and the racks are marked with the GE-logotype.
In the Cassette tray, the cassette positions are marked 1 to 6.
Rack for 250 ml bottlesRack for 50 ml tubesCassette tray
ÄKTA pure User Manual 29119969 AB 97
3 ÄKTA pure external modules
3.2 Fraction collector F9-C
3.2.3 Cassettes, Cassette tray and racks
Note:
Note:
The tray and racks areinserted into the fraction collector with the GE-logotypefacing outwards.
Do not use the Cassette tray when a rack for tubes or bottles is placed in the fraction collector.
Illustration of Cassettes on the Cassette tray
The illustration below shows Cassettes placed on the Cassette tray.
DescriptionPart
Cassette tray1
Cassette position number2
Cassette3
Cassette type code4
Tubes or deep well plates placed in a Cassette5
98 ÄKTA pure User Manual 29119969 AB
QuickRelease function
The Cassettes for the smaller tube sizes (3, 8, and 15 ml) have a built-in QuickRelease function. The
QuickRelease function enables easy handling of tubes in the Cassettes. With the QuickRelease device
in lockposition the tubesare fastened in theCassetteand can easilybe emptied. With the QuickRelease
function in release position, the Cassette can easily be loaded with tubes and used tubes can easily
be discarded.
ActionStep
Load the Cassette with tubes before fractionation:1
•
•
3 ÄKTA pure external modules
3.2 Fraction collector F9-C
3.2.3 Cassettes, Cassette tray and racks
Pull the QuickRelease device to the release position.
Load theCassette with tubes,and press theQuickRelease device tothe locked position.
Locked positionRelease position
3
After fractionation, pull the QuickRelease device to the release position and remove the
tubes containing the fractions of interest.
ÄKTA pure User Manual 29119969 AB 99
3 ÄKTA pure external modules
3.2 Fraction collector F9-C
3.2.3 Cassettes, Cassette tray and racks
ActionStep
Empty and discard the remaining tubes:4
•
Press the QuickRelease device to the lock position, and empty the remaining tubes.
•
Pull the QuickRelease device to the release position, and discard the tubes.
Fraction collector tubes and bottles
The tubes and bottles used in Fraction collector F9-C must fulfill the requirements listed in the table
below. Examples of manufacturers are also listed in the table.
size (ml)
250 ml bottle
L: 55
W: 55
1
W: 64
Discard the tubesEmpty the tubes
Height (mm)Diameter (mm)Tube or bottle
Examples of manufacturers
Max.Min.Max.Min.
NUNC™565011.510.53
VWR™76701210.55
BD™ Biosciences, VWR1029613.3128
BD Biosciences120114171615
BD Biosciences116110302850
Nalgene™, Kautex™121-L: 64.5
1
1
Length and width of the rectangular bottle base
100 ÄKTA pure User Manual 29119969 AB
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