Speed: Increase your productivity.
Scalability: achieve direct, dependable scalability while systematically anticipating errors.
Security: reliable results with sample security.

ÄKTA avant is available in two versions; ÄKTA avant 25 is optimized for media screening and method optimization using small columns. ÄKTA avant 150 is designed for scaling up to larger columns, as well as fine tuning and robustness testing of the optimized process.

Speed

Obtain maximum information from a minimum number of experiments in an automated approach using the integrated Design of Experiments (DoE) functionality. Automated on-line buffer preparation using BufferPro enables easy screening for optimal buffer compositions and quick preparation of single buffers.

Scalability

Scale processes quickly and predictably from screening and method optimization using HiScreen columns to fine-tuning and robustness testing on AxiChrom columns. ÄKTA avant is designed to allow you to take full advantage of modern high flow rate BioProcess media like MabSelect and Capto.

Security

Trace column usage and ensure operational security by using UniTag labels for column recognition. Protect samples from contamination and ensure sample integrity using the cooled, integrated fraction collector.

UNICORN 6

ÄKTA avant is operated by UNICORN 6 control software, which has been specially developed to increase productivity and efficiency.

Videos

ÄKTA avant Overview

ÄKTA avant - Speed, Security & Scalability

ÄKTA avant - System Orientation

ÄKTA avant 25 — Technical Specifications

Complete Packsize	1 Piece
Order Information	Includes two default cassettes for fractionation. Up to six cassettes can be placed in the fraction collector for full capacity. Note, ÄKTA avant is not for use with fraction collector Frac-950. For ordering information, please contact your local GE Healthcare representative.
Application	Fast and secure process development and method optimization
Flow rate	0.001 ml/min-25 ml/min
Operating Pressure Max.	20 MPa (2900 psi)
Operating Pressure [Sample Pump] Max.	10 MPa (1450 psi)
Tubing i.d. [Flow Path]	0.75 mm
Multiple Wavelength Detection	Yes, up to 3 wavelengths
UV Wavelength	190-700 nm
Flow Cell Path Length	2 mm (optional 10 mm)
pH Monitoring	0-14
Multiple Sample Injection	Yes, up to 7 samples
Autosampler Injection	No
Automatic Buffer Preparation	BufferPro
Buffer Selection	18 buffer inlets, option up to 46
Multi-Step Purification	No
Relative Humidity	20%-95% (non condensing)
pH Stability Long Term	1-13
pH Stability Short Term	1-14
Viscosity	0.35 to 10 cP (system pump), 0.7 to 10 cP (sample pump)
Width	860 mm
Height	660 mm
Depth	710 mm
Voltage	100-120/220-240 VAC
Power Consumption	800 VA
Protection Class	IP 21, wet side IP 22
Pump Type	Piston pump, metering type
Compliance	The product fulfills valid directives and standards when used within the conditions specified in the user manual. The product must also be used in the same state as it was delivered from GE Healthcare and connected only to other CE labeled GE Healthcare modules or other products as recommended. For regulatory details please see declaration of conformity.

Application note
Fast process development of a single-step purification using ÄKTA avant systems Summary This study demonstrates a rational strategy for rapid process development of a single-step purification. First, initial screening of chromatography media and process conditions was performed using prefilled PreDictor™ 96-well filter plates and Assist software. Next, the Design of Experiments (DoE) functionality, incorporated in UNICORN™ 6 control software, was used with ÄKTA™ avant 25 system to perform a DoE optimization of loading and elution conditions. DoE was also used to confirm that the optimized process conditions were robust. After optimization, scale-up was performed in two steps (40-fold and 400-fold) with HiScale™ and AxiChrom™ columns using ÄKTA avant 150 system. Close
High-throughput screening and process development for capture of recombinant pro-insulin from E. coli Summary describes a complete process development workflow from PreDictor™ plate screening to a 400 mL pilot-scale purification for capturing recombinant pro-insulin expressed in E. coli. PreDictor 96-well prefilled filter plates, Close
Process scale-up from HiScreen to HiScale columns Summary In this study two capture purification steps were scaled up from HiScreen to HiScale columns in order to evaluate process reproducibility and robustness. The two media used, MabSelect SuRe™ and Capto™ Q showed strong reproducibility and high yield (> 96%) during scale-up, demonstrating the excellent performance and consistency of HiScreen and HiScale columns as well as the BioProcess™ media. Close
Rapid method development for native protein purification using ÄKTA™ avant 25 chromatography system Summary ÄKTA avant 25 system controlled by UNICORN™ 6 software was used to develop a three-step chromatography method for purification of native maltodextrin-binding protein from E. coli. Anion-exchange chromatography (AIEX) was used for the initial capture step, hydrophobic interaction chromatography (HIC) for intermediate purification and gel filtration (GF) for final polishing. HiScreen™ columns were used in a Design of Experiments (DoE) setup to determine optimal loading conditions for capture. In developing the capture step, salt concentration and pH during equilibration, wash, and elution were controlled automatically by on-line buffer preparation using BufferPro. Screening of chromatography media for the intermediate purification step was performed using HiTrap™ HIC Selection Kit. Finally, polishing by GF ensured removal of remaining impurities resulting in target protein purity of almost 100%. After optimization of the capture step and screening HIC media for the intermediate step, the whole purification process was performed in a single day. Close
Rapid process development for purification of a MAb using ÄKTA™ avant 25 Summary ÄKTA avant 25 system controlled by UNICORN™ 6 software was used to develop a two-step chromatography process for purification of a monoclonal antibody (MAb). MabSelect SuRe™, a protein A-based chromatography medium (resin) was used for the initial capture step while the multimodal anion exchanger Capto™ adhere was used for reduction of impurities in a second, polishing step. An experimental design was applied to the polishing step to screen loading conditions. Sample pH, conductivity, and load were varied. Using 1 ml prepacked HiTrap™ columns, the design was run in less than 24 h and a reduced design was performed to establish the robustness of the process conditions. Using the Design of Experiments (DoE) functionality incorporated in UNICORN 6 together with HiScreen™ prepacked columns, optimization of the overall process was achieved in approximately one week. Despite the challenging nature of the feed studied, high yield and purity of the target MAb was achieved. Close

Brochure
Your protein is unique Summary Proteins are as different as you and me. When you need to purify them, one size definitely does not fit all. This is where ÄKTA™ avant excels - by efficiently and precisely purifying any protein with the individual respect it deserves. Close
ÄKTA™ Protein purification by design Summary With the ability to purify virtually any biomolecule, the ÄKTAdesign platform can handle the simplest and the toughest of challenges. It gives you speed, ease of use and flexibility whatever your purification application or scale. The platform covers all major chromatographic and cross flow filtration techniques, from the research laboratory to process development and manufacturing. Systems in the ÄKTAdesign platform work with intelligent UNICORN™ software, which makes it simple to control every stage of your purification process. A broad range of prepacked columns, media and filters provides more options for the best results. Close
ÄKTA™ avant - Inspired to Äccelerate your process development Summary ÄKTA is the Swedish word for true or real. And it is the name for GE Healthcare systems that have enabled over 100,000 users to achieve breakthroughs and advances within protein purification and separation. This proud lineage now sky rockets into the next generation with ÄKTA avant. ÄKTA avant is the result of research and close dialog with you. Your realities and needs have been the source of our inspiration. We have listened, learned, and developed one system encapsulating features and functions that work smoothly together to allow you to achieve your goals. ÄKTA avant allows fast and secure development of scalable chromatographic processes for biopharmaceutical production. Close

Data file
UNICORN™ 6.2 control software Summary UNICORN system control software provides built-in knowledge for planning, controlling, and analyzing chromatography systems and results. UNICORN is used on all ÄKTA™ design systems and can be used throughout all stages of process development from the lab bench to full-scale production. UNICORN 6 control software has been specially developed for use with ÄKTA avant liquid chromatography system. ÄKTA avant, together with UNICORN 6, enables you to achieve fast and secure process development. UNICORN 6 features a user friendly graphical user interface (GUI), an improved Method Editor, and an integrated Design of Experiments (DoE) tool. Close
ValidationCare Summary Datafile describing the content and benefit of our new Validation Care Service. Validation Care is a NPI within Validation Support, a IQ/OQ + GxP validation service contract to be sold through our sales force, targeting pharma customers in a regulated environment having the following systems from GEHC; ÄKTA Pilot, ÄKTA Avant, ÄKTA Explorer and later also ÄKTA Process* . Close

Data file

UNICORN™ 6.2 control software Summary

ValidationCare Summary

Handbook
ÄKTA Laboratory-scale Chromatography Systems - Instrument Management Handbook Summary This handbook is focused on liquid chromatography systems used for protein purification at research laboratory scale. Beginners can use the handbook to obtain an overview of how purification systems work and to learn about important considerations for achieving successful results. Experienced system users will also find valuable and detailed information on different hardware modules. Close

Handbook

ÄKTA Laboratory-scale Chromatography Systems - Instrument Management Handbook Summary

Installation guide
UNICORN 6.x - SQL Server 2008 configuration for UNICORN installations Summary This document describes the necessary settings when preparing a Microsoft™ SQL Server 2008 database installation for use with UNICORN 6.x. By default, a Microsoft SQL Express 2008 database is installed as part of the UNICORN installation. However, if a Microsoft SQL Server 2008 database server installation is preferable, it is possible to install UNICORN 6.x onto that server instead. However, some prerequisite settings must be made for the server before UNICORN is installed. These settings are described below. If nothing else is noted, the settings are made in the SQL Server 2008 Setup wizard. Wizard steps which are not described below can be bypassed without changes by clicking the Next button. This document is primarily intended for GE Healthcare field service personnel, as well as IT administrators of large customers, as a complement to the regular UNICORN 6.x documentation. Close
UNICORN™ 6.0 to 6.1 Post upgrade instruction Summary This document describes steps that may be necessary to perform to ensure proper operation after an upgrade of UNICORN 6.0 to UNICORN 6.1. Specifically, the instruction describes a repair of the OPC Core Components. This document is primarily intended for GE Healthcare field service personnel, as a complement to the regular UNICORN 6.1 documentation. Close

Installation guide

UNICORN 6.x - SQL Server 2008 configuration for UNICORN installations Summary

UNICORN™ 6.0 to 6.1 Post upgrade instruction Summary

Instruction/protocol
Rack, for 50 ml tubes - Rack, for 250 ml bottles Summary This document contains information about Rack for 50 ml tubes and Rack for 250 ml bottles, for use with ÄKTA avant 25 and 150. Close
Spare parts recommended to keep on site for ÄKTA avant 25 Close
Wetted materials ÄKTA avant Close
ÄKTA avant Information Letter Summary Under normal operating conditions, ÄKTA avant will perform as expected and with few problems. However, certain exceptional conditions may impair the performance and may require special procedures in order to resolve the issue. This document includes additional advice and tips for issues regarding: • System • Fraction collector • BufferPro • Intelligent Packing (only available for ÄKTA avant 150) Close

Magazine
Discovery Matters 10 Summary Discovery Matters is our customer magazine targeted to R&D customers in academia, biotechnology, and pharma companies. Discovery Matters 10 covers new products launched from in Q2 and Q3 2009, technical tips, a technology central feature, and scientific articles. The main feature of this issue is content relating to the launch of a new ÄKTA™ system, ÄKTA avant 25 and its companion product, UNICORN™ 6. The article section includes material on fast purity analysis using Superdex™ short gel filtration columns, a feature on ÄKTAmicro™ system for purification of low-abundant proteins, using WAVE Bioreactor system for culture of adherent cells, and much more. Close
Upstream and Downstream issue 3 Summary Upstream and Downstream is GE Healthcare's magazine targeted to customers in the biopharmaceutical industry. Issue 3 (November 2010) contains the following articles: 1) Fast and dependable scalability in process development with ÄKTA™ avant 150, UNICORN™ 6.1, and HiScale™ columns 2) Manufacturing Solutions to enhance filtration and chromatography productivity 3) High-throughput screening methods to speed MAb process development 4) Faster scale-up of a two-step MAb process with AxiChrom™ columns and ReadyToProcess™ solutions 5) ReadyCircuit™ and ReadyKart promote fast, flexible bioprocessing setups 6) Overcoming protein stratification in cross-flow filtration using ReadyCircuit 2-D bags 7) Ways to improve vaccine development and manufacture 8) Determining optimal filtration conditions using ÄKTAcrossflow™ and Design of Experiments. Close

Magazine

Discovery Matters 10 Summary

Upstream and Downstream issue 3 Summary

Software
UNICORN 6.1 (or later versions) Pre Defined Columns File Summary When a new list is imported to UNICORN 6.1 (or later versions), it will replace all the predefined column types in the database with the updated column types. Only predefined column types will be replaced. User defined column types, both personal and global, will remain in the database. The table below describes how to import a new column list into the database: Step Action 1 Save the new column list (.zip file). 2 In the Column Type Parameters tab in the Column Handling dialog in the Method Editor, click the Import button. Result: The Import dialog opens. 2 Locate the zip file with the column list to be imported and click Open. Result: The Import confirmation dialog opens, explaining what will happen when the zip file is imported. 3 Click Yes. Result: The new list of predefined column types is imported into the database. Close
ÄKTA avant Instrument Configurations Summary Description These configurations apply to: • ÄKTA avant 25 • ÄKTA avant 150 UNICORN™ 6.1 is required before the Instrument configuration can be installed. Contact your local GE Healthcare representative for UNICORN software update. IMPORTANT! Read the Installation instruction (28990410AE.pdf) before you download one of the zip files. Download the appropriate zip file to your computer: AKTAavant25_2.3.1.0.zip if you use ÄKTA avant 25 AKTAavant150_1.4.0.0.zip if you use ÄKTA avant 150 Close

Software

UNICORN 6.1 (or later versions) Pre Defined Columns File Summary

ÄKTA avant Instrument Configurations Summary

Software change description
UNICORN™ 6.1 vs UNICORN 6.0 Software Change Description Summary This document describes the implemented changes and improvements in UNICORN 6.1compared to version 6.0. Close

Software information letter
UNICORN 6.1 Information Letter Summary This Information letter contains descriptions of different scenarios that may occur when UNICORN 6.1 is used. If applicable, actions to avoid or correct problems resulting from the described scenarios are suggested. This information is valid for UNICORN 6.1 software (build 6.1.0.495) and ÄKTA™ avant instruments only. Close

User manual
UNICORN™ 6 Summary The purpose of the UNICORN Method Manual is to provide a comrehensive guide to creating methods that can be run on an ÄKTAavant system. It covers the features and tools included in the Method Editor module of the UNICORN software with practical instructions. The manual covers the following: • how to create methods and phases. • how to use BufferPro. • how to design and optimize experiments using Design of Experiments and Scouting. • how to use method queues. • how to handle column types and individual columns. For advanced users, an overview of how to edit methods at the level of individual instructions is also given. Note: The Method Manual does not describe the functions of every command in all panes and dialogs of the user interface. Refer to the online help for information about commands that are not described in this manual. The online help in the Method Editor module is accessed either by clicking Help buttons in software dialogs, by pressing the F1 key, or selecting Help:Help for Method Editor. Close
UNICORN™ 6 Administration and Technical Manual Summary The UNICORN 6 Administration and Technical Manual describes how to install, set up and maintain the UNICORN software. For the most part, the installation is assumed to be in a network environment. A system administrator should be able to refer to this manual for the information that is necessary to ensure that the network and all UNICORN clients operate smoothly. Ordinary users, not assigned as system administrators, may need to refer to parts of this manual. This is explained below. Close
UNICORN™ 6 Advanced system administration Summary The UNICORN 6 Advanced system administration manual describes settings and procedures necessary for operation mainly in a network environment. This manual covers topics that are not included or comprehensively described in the UNICORN 6 Administration and Technical Manual (28944078AC). It should be used as a reference material for system administrators. Close
UNICORN™ 6 Evaluation Manual Close
UNICORN™ 6.1 Administration and Technical Manual Summary The UNICORN 6.1 Administration and Technical Manual provides the necessary information for the UNICORN system administrator. Topics covered include installation and configuration, network and database administration, user account, workstation and server set up. Close
UNICORN™ 6.1 Evaluation Manual Summary The UNICORN Evaluation Manual provides comprehensive information about how to evaluate results from separation runs performed using UNICORN software. The manual also describes how to create and edit reports. Close
UNICORN™ 6.1 Method Manual Summary The UNICORN Method Manual provides comprehensive information about how to create and edit UNICORN methods. This manual also includes information about Design of Experiment using UNICORN 6.1. Close

Technical Support

Find your local representative contact information

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FAQ

Frequently Asked Questions.

Product Related FAQs

Why should I have REGULAR, PLANNED MAINTENANCE on my system?

How can I USE my system in COLD ROOMS?

How can I clean my ÄKTA avant system?

How can I CLEAN my system?

ÄKTA avant

Cleaning the system
The procedures described below are for system cleaning.

1. Clean the UV flow cell

Maintenance interval
Clean the UV flow cell every six months, or when required.
NOTICE: Keep UV flow cell clean. Do not allow solutions containing dissolved salts, proteins or other solid solutes to dry out in the flow cell. Do not allow particles to enter the flow cell, as damage to the flow cell may occur.

Required material
The following material is required:
• Luer connector
• Waste container
• Syringe, 25-30 ml
• 10% surface active detergent solution (e.g., Decon 90, Deconex 11, or RBS 25)
• Distilled water

Instruction
Follow the instruction below to clean the UV flow cell. The UV flow cell can be either mounted or not mounted on the instrument during the cleaning procedure.

Step	Action
1	Disconnect the fingertight connector and the tube from the top of the UV flow cell, and replace the fingertight connector with a Luer connector.

2	Disconnect the tube from the bottom of the UV flow cell, and connect a waste tube to the UV flow cell. Immerse the waste tube in a waste container.
3	Fill a syringe with distilled water, and connect the syringe to the Luer connector.

4	Squirt the distilled water through the UV flow cell in small amounts. Disconnect the syringe.
5	Fill a syringe with a 10% surface active detergent solution like Decon 90, Deconex 11, RBS 25 or equivalent, and connect the syringe to the Luer connector.
6	Squirt the detergent solution through the UV flow cell about five times.
7	Leave the detergent solution in the flow cell for at least 20 minutes.
8	Inject the detergent solution remaining in the syringe into the flow cell. Disconnect the syringe.
9	Fill a syringe with distilled water. Connect the syringe to the Luer connector.
10	Inject the distilled water into the UV flow cell to rinse the flow cell. Disconnect the syringe.
11	Disconnect the Luer connector from the top of the UV flow cell, and reconnect the fingertight connector with the tube.
12	Disconnect the waste tubing from the bottom of UV flow cell. Reconnect the piece of tubing from the Conductivity monitor to the bottom of the UV flow cell.

2. Clean the instrument externally

Maintenance interval
Clean the the instrument externally when required. Do not allow spilled liquid to dry on the instrument.

Required material
The following material is required:
• Cloth
• Mild cleaning agent or 20% ethanol

Instruction
Follow the instruction below to clean the instrument externally.

Step	Action
1	Check that no run is in progress.
2	Switch off the instrument.
3	Wipe the surface with a damp cloth. Wipe off stains using a mild cleaning agent or 20% ethanol. Wipe off the excess.
4	Let the instrument dry completely before using it.

3. Perform System CIP, Cleaning-in-place

Maintenance interval
Perform a System CIP when required, for example between runs where different samples and buffers are used. This is important to prevent bacterial growth in the instrument.

Required material
The following material is required:
• Appropriate cleaning solutions (e.g., NaOH, buffer solution or distilled water).
• Syringe, 25-30 ml

Instruction
Follow the instruction in Section 7.6.2 Perform System CIP in the ÄKTA avant and UNICORN 6.1 User Manual.

4. Clean the Fraction collector

Maintenance interval
Clean the Fraction collector when required, for example if liquid has been spilled in the Fraction collector chamber.

Required material
The following material is required:
• Wash bottle
• Water or 20% ethanol
• Cloth

Instruction
Follow the instruction below to clean the interior of the Fraction collector. See the locations of the components of the Fraction collector in Section 3.4.1 Illustrations of the Fraction collector, in the ÄKTA avant and UNICORN 6.1 User Manual.

Step	Action
1	In System Control, select Manual:Execute Manual Instructions:Fraction collection:Frac cleaning position. Click Execute. Result: The Dispenser head moves to cleaning position, and the Instrument display states System pause.
2	Open the Frac drawer and lift off the Cassette tray.
3	Wash the Cassettes and the Cassette tray with water and a mild cleaning agent.
4	Lift off the Waste funnel and wash it with water and a mild cleaning agent. Refit the Waste funnel.
5	In ÄKTA avant 150 only: Remove the Dispenser head cover and wash it with water and a mild cleaning agent. Refit the Dispenser head cover.
6	Wipe off the Frac chamber using a damp cloth. Wipe off stains using a mild cleaning agent or 20% ethanol. Note: Be careful not to damage the Arm flat cable. TIP: The Frac arm can easily be moved to facilitate cleaning of the Fraction collector. TIP: The Cassette tray positioning discs can easily be removed to facilitate cleaning of the Fraction collector. Make sure to refit the discs before the Cassette tray is inserted in the Frac chamber.
7	Wipe off the Dispenser head and its diode windows using a wash bottle with water or 20% ethanol and a cloth. ÄKTA avant 25 has four diode windows and ÄKTA avant 150 has two diode windows (see the illustrations below). Note: Be careful not to scratch the diode windows.
8	Let the Fraction collector dry completely before starting a run.
9	Close the Frac drawer. Result: Automatic scanning is performed.
10	In the System Control module, press the End icon in the toolbar. Result: The Dispenser head moves to home position, and the Instrument display states Ready.

5. Clean the pH electrode

Maintenance interval
Clean the pH electrode when required. During cleaning, the pH electrode can be either installed in the pH valve, or not installed in the pH valve. The pH electrode has a limited longevity and should be replaced every six months or when the response time is slow, see Section 7.5.2 Replace the pH electrode, in the ÄKTA avant and UNICORN 6.1 User Manual. After cleaning has been performed, re-calibrate the pH monitor, see Section 7.7.1 Calibrate the pH monitor, in the ÄKTA avant and UNICORN 6.1 User Manual.

Required material
• Syringe, 25-30 ml
• Distilled water
• 0.1 M HCl and 0.1 M NaOH

• Liquid detergent

• 1% pepsin solution in 0.1 M HCl

• 1 M KNO₃

Cleaning agents
Clean the pH electrode using one of the following procedures:

Salt deposits
Dissolve the deposits by immersing the electrode in:
• 0.1 M HCl
• 0.1 M NaOH
• 0.1 M HCl
each for five minutes. Rinse electrode tip in distilled water between each solution.

Oil or grease films
Wash the electrode tip in liquid detergent and water. If the films are known to be soluble in a particular organic solvent, wash with this solvent. Rinse the electrode tip in distilled water.
Protein deposits
Dissolve the deposit by immersing the electrode in a 1% pepsin solution in 0.1 M HCl for five minutes followed by thorough rinsing with distilled water. If these procedures fail to rejuvenate the electrode, try the following procedure.

Note: This procedure can be performed only when the pH electrode is not installed in the pH valve.

Step	Action
1	Heat a 1 M KNO₃ solution to 60°C–80°C.
2	Place the electrode tip in the heated KNO₃ solution.
3	Allow the electrode to cool while immersed in the KNO₃ solution before re-testing.

If these steps fail to improve the electrode, replace the electrode, see Section 7.5.2 Replace the pH electrode, in the ÄKTA avant and UNICORN 6.1 User Manual.

6. Clean a pH electrode installed in the pH valve

Follow the instruction below to clean a pH electrode installed in the pH valve. The calibration function is used to switch the pH valve. However, no calibration is performed.

Step	Action
1	Open the System Control module and select System:Calibrate. Result: The Calibration dialog opens.
2	Set Monitor to calibrate by selecting pH from the list.
3	Press the Prepare for calibration button. Result: The pH valve switches to the calibration position.
4	Fill a syringe with approximately 10 ml of chosen cleaning solution. Connect the syringe to the pH valve port Cal. Inject the liquid and wait for 5 minutes. Disconnect the syringe.
5	If several cleaning solutions are to be used, repeat step 4 with distilled water and then with the next solution.
6	As the last step in the cleaning procedure: • Fill a syringe with distilled water. • Connect the syringe to the pH valve port Cal. • Inject the water. • Disconnect the syringe.
7	Press the Close button.
8	Result: The pH valve switches back to the default position and the Calibration dialog closes. No calibration is performed.

7. Clean the check valves of a pump head

Maintenance interval
Clean the check valves when required, for example if solids in the check valve cause irregular or low flow.
Required material
The following material is required:
• Adjustable wrench
• Methanol
• Distilled water
• Ultrasonic bath

Instruction
Follow the instruction below to remove and clean the check valves of the Sample pump or the system pumps.

Step	Action
1	Switch off the instrument.
2	Disconnect the tubing from the pump head and disconnect the pump inlet tubing. Disconnect the tubing of the pump rinsing system.
3	Unscrew the purge valve by turning it counter-clockwise, and lift off the metal ring.
4	Unscrew the plastic nut of the upper check valve using an adjustable wrench, and gently lift off the upper check valve.
5	Unscrew the two white plastic screws located below each pump head. Pull the plastic connectors to the sides to release the inlet manifold.
6	Unscrew the lower check valve using an adjustable wrench.
7	WARNING: Hazardous substances. When using hazardous chemical and biological agents, take all suitable protective measures, such as wearing protective glasses and gloves resistant to the substances used. Follow local and/or national regulations for safe operation and maintenance of the system. Immerse the complete valves in methanol and place them in an ultrasonic bath for a few minutes. Repeat the ultrasonic bath with deionized water.
8	Refit the check valves. Follow the instructions below to fasten the cleaned check valve: • For black plastic valves: Tighten the nut until fully finger-tight and then use the adjustable wrench to tighten a further 90 degrees. • For metal valves: Tighten the nut until fully finger-tight and then use the adjustable wrench to tighten a further 90 degrees.
9	Refit the inlet manifold and reconnect the tubing to the pump head.

8. Clean the conductivity flow cell

Maintenance interval
Clean the Conductivity flow cell when required.

Required material
The following material is required:
• Luer connector
• Waste container
• Syringe, 25-30 ml
• 1 M NaOH
• Distilled water

Instruction
Follow the instruction below to clean the flow cell of the Conductivity monitor.

Step	Action
1	Disconnect the fingertight connector and the tube from the top of the Conductivity monitor, and attach a Luer connector.
2	Disconnect the tube from the bottom of the Conductivity monitor, and connect a waste tube to the Conductivity monitor. Immerse the waste tubing in a waste container.
3	Disconnect the Conductivity monitor from the rails, or slide the Conductivity monitor to the right on the rails.
4	Fill a syringe with distilled water, and connect the syringe to the Luer connector.
5	Squirt distilled water through the Conductivity flow cell in small amounts. Disconnect the syringe.
6	WARNING: Hazardous substances. When using hazardous chemical and biological agents, take all suitable protective measures, such as wearing protective glasses and gloves resistant to the substances used. Follow local and/or national regulations for safe operation and maintenance of the system. Fill a syringe with 1 M NaOH, and connect the syringe to the Luer connector.
7	Squirt 1 M NaOH through the Conductivity flow cell about five times.
8	Leave the liquid in the flow cell for at least 15 minutes.
9	Fill a syringe with distilled water. Connect the syringe to the Luer connector.
10	Inject the distilled water into the Conductivity flow cell to rinse the flow cell. Disconnect the syringe.
11	Disconnect the Luer connector from the top of the Conductivity flow cell, and reconnect the fingertight connector with tubing.

How can I TEST THE PERFORMANCE of my system?

How can I determine the delay volume?

How can I determine the delay volume?

Three different methods are described below:

Method I - Determining the delay volume of your system by measuring different retention volumes

1) Check that the pump is delivering the correct flow at 1 ml/min. If the measured flow rate differs the retention volumes need to be corrected.

2) Mount a small loop, e.g. 100 µl, and prepare a solution of 5% acetone in water to use as sample.

3) Fill the system with clean water. Run manually or make your own method. Start the pump at 1 ml/min and inject the sample. The measured retention volume is called Volume 1.

4) Re-configure your system. Dismount the tubing from the UV flow cell and insert a low dead volume connector, e.g. a female-female Valve connector, in the flow path (replaces the UV flow cell). Mount the “tubing end” of the frac outlet into the UV flow cell and mount a waste-tubing to the bottom of the UV flow cell.

5) Set Frac size to a very large volume, e.g. 100 ml, so that the valve is in the Frac position during the entire run. Start the pump at 1 ml/min and inject the sample. The measured retention volume is called Volume 2. Delay volume = Volume 2 – Volume 1.

Method II - Determining the delay volume of your system with the help of another ÄKTA system

This method is more accurate than the one described above, but demands one more system.

1) Check that the pump on your chosen ÄKTA system is delivering the correct flow at 1 ml/min. If the measured flow rate differs the retention volumes need to be corrected.

2) Mount a small loop, e.g. 100 µl, and prepare a solution of 2-5% acetone to use as sample.

3) Fill the system with clean water. Run manually or make your own method. Start the pump at 1 ml/min and inject the sample. The measured retention volume is called Volume 1.

4) Re-configure your system. In e.g. an ÄKTAprime system dismount the tubing from the UV flow cell to the Fraction collector including the restrictor, the Cond cell and the Frac valve. In the other ÄKTA system, e.g. ÄKTApurifier or ÄKTAexplorer, mount the “prime tubing kit” between the tubing to the UV flow cell and the UV flow cell. To do this a low dead volume connector, e.g. a female-female Valve connector, is needed.

5) In order to have the correct flow path the Frac valve in the “prime tubing kit” must be activated. This can be achieved be connecting it to the Valve B port of the P-900 pump in the ÄKTApurifier or ÄKTAexplorer system. The valve is activated by setting Pump B inlet to the B2 position.

6) Start the pump at 1 ml/min and inject the sample. The measured retention volume is called Volume 2. Delay volume = Volume 2 – Volume 1.

Method III - Determining the delay volume by balancing eluted water

Manually set the flow path to the direction of the fraction collector. Unscrew the tubing that is connected to inlet of the UV flow cell and insert a luer adaptor instead. Fill a syringe with water and inject water into flow cell unless it drops at the outlet of the fraction collector (in which case you have likely exceeded the pressure in the tubing which might be more than 4 bar, depending on configuration and flow restrictor used). Now fill the syringe with air (at least 20 ml because of compression) and displace the water. Collect eluting water in a small cup. Determine the system delay volume by balancing the cup before and after elution. Repeat two times for calculation of a mean value. Enter the mean value in "system settings" in UNICORN.

Which EMPTY COLUMNS are recommended for my ÄKTA avant system?

ÄKTA avant 25	ÄKTA avant 150
Columns	Columns
Tricorn 5, High Performance Column	HiScale 16
Tricorn 10, High Performance Column	HiScale 26
HiScale 16	HiScale 50
HiScale 26	XK 16
HiScale 50	XK 26
XK 16	XK 50
XK 26	AxiChrom 50
XK 50	AxiChrom 70

I want to know the DIMENSIONS and WEIGHT of my system.

I want to know the FLOW and PRESSURE RANGE of my system?

What kind of ORGANIC SOLVENT can I use with my ÄKTA avant system?

What kind of ORGANIC SOLVENT can I use with my ÄKTA avant system?

ÄKTA avant

Chemical resistance guide and chemical compatibility
The chemical resistance of ÄKTA avant to some of the most commonly used chemicals in liquid chromatography is indicated in the table below.

The ratings are based on the following assumptions:
1. The synergistic effects of the chemical mixtures have not been taken into account.
2. Room temperature and limited over-pressure is assumed.

Note: Chemical influences are time and pressure dependent. Unless otherwise stated, all concentrations are 100%.

Usage	Name	Concentration	CAS no and EEC no
Separation	Aqueous buffers, pH 2-12
	Acetic acid	5%
Cleaning-In-Place (CIP)	Acetic acid	70%	64-19-7 200-580-7
Reversed Phase Chromatography (RPC)	Acetonitrile Remarks: Quaternary valve not resistant. Depending on pressure, tubing between pump head, and pressure monitor needs to be changed. See section Prepare inlet tubing, in the User Manual.	100%	75-05-8 200-835-2
RPC	Acetonitrile/Tetrahydrofuran (THF) Remarks: Quaternary valve not resistant.	85/15	109-99-9 (Tetrahydrofuran) 203-726-8
RPC	Acetone	10%
RPC	Acetone	100%	67-64-1 200-662-2
Oligonucleotide synthesis	Ammonia	30%	7664-41-7 231-635-3
Hydrophobic Interaction Chromatography (HIC)	Ammonium chloride	Saturated	12125-02-9 235-186-4
Purification of plasmids	Ammonium sulphate	4 M	7783-20-2 231-984-1
Wash, using Protein A gels, Refolding	Arginine	2 M	74-79-3 200-811-1
Cleaning and storage of columns	Benzyl alcohol	2%	100-51-6 202-859-9
HIC	Citric acid	100%	29340-81-6 249-576-7
Cleaning	Decon 90	10%
RPC, CIP, Cell separation	Dimethyl sulphoxide (DMSO)	5%	67-68-5 200-664-3
Cleaning, Plasmid separation	Dimethylformamide (DMF) Remarks: Quaternary valve not resistant	100%	1968-12-02 200-679-5
Reducing agent	Dithiothreitol (DTT)	0.1 M	4413-31-4 NA
Reducing agent	DTE	0.1 M
Buffer additive	Ethylenediaminetetraacetic acid (EDTA)	100%	60-00-4 200-449-4
CIP	Ethanol	100%	75-08-1 200-837-3
Storage	Ethanol/Acetic acid	20%/10%
HIC	Ethylene glycol	100%	107-21-1 203-473-3
HIC	Ethylene glycol	100%
RPC	Formic acid	100%	64-18-6 200-579-1
HIC	Glycerol	100%	56-81-5 200-289-5
Denaturing of proteins	Guanidinium hydrochloride	6 M
Cleaning of MAb binding media	Glycine	2 M	56-40-6 200-272-2
CIP	Hydrochloric acid	0.1 M	7647-01-0 231-595-7
CIP	Hydrochloric acid	2 M
Buffer preparation	Imidazole	1 M	288-32-4 206-019-2
CIP	Isopropanol	100%	67-63-0 200-661-7
RPC, CIP	Methanol	100%	74-93-1 200-659-6
Reducing agent	Merkaptoethanole	10%	37482-11-4 253-523-3
RPC	n-Propanol	100%	67-63-0 200-661-7
CIP	Phosphoric acid	10%	7664-38-2 231-633-2
	Potassium carbonate	Saturated	584-08-7 209-529-3
pH cell storage	Potassium chloride	Saturated	7447-40-7 231-211-8
Detergent	Sodium dodecyl sulfate (SDS)	10%	151-21-3 205-788-1
	Sodium chloride	Saturated	7647-14-5 231-598-3
CIP	Sodium hydroxide	2 M	1310-73-2 215-185-5
CIP	Sodium hydroxide	1 M, 50°C
	Sodium hydroxide + sodium hypochloride	(1.0 M+500 ppm), 50°C
HIC	Sodium sulphate	Saturated	7757-82-6 231-820-9
	Trichloroacetic acid	1%	76-03-9 200-927-2
	Trifluoroacetic acid	1%	176-05-1 200-929-3
Detergent	Triton-X	1%	9002-93-1 (Triton X 100)
Detergent	Tween	1%	9005-64-5 (Tween 20) 500-018-3
Buffer additive	Urea	8M	57-13-6 200-315-5
	Water	100%	7732-18-5 231-791-2

What DOCUMENTS are supplied with AKTA avant system?

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Flow path schemes

System flow path description.

ÄKTAavant 25 and ÄKTAavant 150

Tubing, Flow path scheme ÄKTAavant 25

Tubing description ÄKTAavant 25

ÄKTAavant 25 and ÄKTAavant 150

Illustration of the liquid flow path
The illustrations below show detailed flow charts for ÄKTAavant 25 and 150. The flow charts show interconnections between instrument components. The blue labels refer to tubing, in the User Manual.

ÄKTAavant 25

ÄKTAavant 150

UNICORN flow scheme
During a run, the current liquid flow path is displayed in the Flow Scheme pane of the SystemControl module in UNICORN. Open flow paths are displayed in green and closed flow paths are displayed in grey. Red modules indicate alarms. Real-time data from monitors are also shown in the flow scheme.
To view and/or issue instructions controlling a module, double-click the module in the flow scheme or right-click the module and select Instructions and the instruction of interest. The Manual Instructions dialog opens with the chosen instruction selected.
To view the ports and the different flow paths of a valve, right-click the valve in the flow scheme and select Show Details. An illustration of ports and available flow paths opens.

Illustration of UNICORN flow scheme
The flow scheme displayed in UNICORN is shown in the illustration below.

Flow paths of Injection valve

The Injection valve can be set to different positions that give different flow paths through the valve.
The illustration and table below describe the different flow paths of the Injection valve.
The method progressing flow, marked in the illustration below, defines the flow from which the volume base is calculated.

#	Product Name	Product Code	Price	Qty

Tubing, Flow path scheme ÄKTAavant 25

Flow path scheme ÄKTAavant 25

#	Product Name	Product Code	Price	Qty

Tubing description ÄKTAavant 25

Inlet tubing

The table below shows the labels, diameters and the standard lengths of the inlet tubing. The inlet tubing is mounted on delivery.

Label	Description	Tubing	Length (mm)
A1 - A7	Inlets to Inlet valve A	PTFE, o.d. 1/8", i.d. 1.6 mm	1500
B1 - B7	Inlets to Inlet valve B	PTFE, o.d. 1/8", i.d. 1.6 mm	1500
Q1 -Q4	Inlets to Quaternary valve	PTFE, o.d. 1/8", i.d. 1.6 mm	1500
Buff	Buffer inlet to Sample inlet valve	PTFE, o.d. 1/8", i.d. 1.6 mm	1000
S1 - S7	Inlets to Sample inlet valve	PTFE, o.d. 1/8", i.d. 1.6 mm	700
InQA	From Quaternary valve to Inlet valve A	PTFE, o.d. 1/8", i.d. 1.6 mm	110
InQB	From Quaternary valve to Inlet valve B	PTFE, o.d. 1/8", i.d. 1.6 mm	110
InA	From Inlet valve A to System pump A	PTFE, o.d. 1/8", i.d. 1.6 mm	220
InB	From Inlet valve B to System pump B	PTFE, o.d. 1/8", i.d. 1.6 mm	220
InS	From Sample inlet valve B to Sample pump	PTFE, o.d. 1/8", i.d. 1.6 mm	220

High pressure tubing

The table below shows the labels, diameters and the standard lengths of the high pressure tubing. The high pressure tubing is mounted on delivery.

Label	Description	Tubing	Length (mm)
1A1	System pump A left to Restrictor A	PEEK, o.d. 1/16", i.d. 0.75 mm	340
1A2	System pump A right to Restrictor A	PEEK, o.d. 1/16", i.d. 0.75 mm	340
2A	Restrictor A to Pressure monitor	PEEK, o.d. 1/16", i.d. 0.75 mm	100
1B1	System pump B left to Restrictor B	PEEK, o.d. 1/16", i.d. 0.75 mm	340
1B2	System pump B right to Restrictor B	PEEK, o.d. 1/16", i.d. 0.75 mm	340
2B	Restrictor B to Pressure monitor	PEEK, o.d. 1/16", i.d. 0.75 mm	100
1S1	Sample pump left to Restrictor S	PEEK, o.d. 1/16", i.d. 0.75 mm	340
1S2	Sample pump right to Restrictor S	PEEK, o.d. 1/16", i.d. 0.75 mm	340
2S	Restrictor S to Pressure monitor	PEEK, o.d. 1/16", i.d. 0.75 mm	100
3	Pressure monitor to Mixer	PEEK, o.d. 1/16", i.d. 0.75 mm	280
3S	Pressure monitor S to Injection valve	PEEK, o.d. 1/16", i.d. 0.75 mm	485
4	Mixer to Injection valve	PEEK, o.d. 1/16", i.d. 0.75 mm	400
5	Injection valve to Column valve	PEEK, o.d. 1/16", i.d. 0.50 mm	180
6	Column valve to UV monitor	PEEK, o.d. 1/16", i.d. 0.50 mm	160
7	UV monitor to Conductivity monitor	PEEK, o.d. 1/16", i.d. 0.50 mm	100
8	Conductivity monitor to pH valve	PEEK, o.d. 1/16", i.d. 0.50 mm	165
9	pH valve to Outlet valve	PEEK, o.d. 1/16", i.d. 0.50 mm	215
1R	To Flow restrictor	PEEK, o.d. 1/16", i.d. 0.50 mm	75
2R	From Flow restrictor	PEEK, o.d. 1/16", i.d. 0.50 mm	75
Frac	Outlet valve to fraction collector	PEEK, o.d. 1/16", i.d. 0.50 mm	1280

Reference capillaries

The table below shows the labels, diameters and the standard lengths of the reference capillaries.

Label	Description	Tubing	Length (mm)
Ref 1	Reference capillary	PEEK, o.d. 1/16", i.d. 0.25 mm	400
Ref 2	Reference capillary	PEEK, o.d. 1/16", i.d. 0.13 mm	700

Outlet tubing

The table below shows the labels, diameters and the standard lengths of the outlet tubing.

Label	Description	Tubing	Length (mm)
Out1 - Out 32	Outlets from Outlet valve	PTFE, o.d. 1/8", i.d. 1.6 mm	1000

Waste tubing

The table below shows the labels, diameters and the standard lengths of the waste tubing. The waste tubing is mounted on delivery.

Label	Description	Tubing	Length (mm)
W1	System pump waste	Tefzel, o.d. 1/16", i.d. 1.0 mm	1900
W2	Sample pump waste	Tefzel, o.d. 1/16", i.d. 1.0 mm	1900
W3	pH valve waste	Tefzel, o.d. 1/16", i.d. 1.0 mm	1900
W	System waste	Tefzel, o.d. 1/16", i.d. 1.0 mm	1900
N/A	Top tray waste	Silicon, o.d. 12 mm, i.d. 8 mm	2500
N/A	Frac front waste	Silicon, o.d. 12 mm, i.d. 8 mm	2100
N/A	Frac back waste	Silicon, o.d. 12 mm, i.d. 8 mm	1600

#	Product Name	Product Code	Price	Qty

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Maintenance

Maintenance instructions and procedures.

ÄKTAavant

Periodic maintenance program

This section lists the periodic maintenance activities that should be performed by the user of ÄKTA avant, as well as maintenance activities that should be performed when required.

	Warning! Hazardous biological agents during run. When using hazardous biological agents, run System CIP and Column CIP to flush the entire system tubing with bacteriostatic solution (e.g., NaOH) followed by a neutral buffer and finally distilled water, before service and maintenance.
	Warning! Hazardous chemicals during run. When using hazardous chemicals, run System CIP and Column CIP to flush the entire system tubing with distilled water, before service and maintenance.
	Warning! Always use appropriate personal protective equipment during operation and maintenance.

Maintenance operations should be performed by the user at regular intervals

Daily Maintenance

Weekly Maintenance

Monthly Maintenance

6 Month Maintenance

When Required

Maintenance when required

Show info

Troubleshooting

Find solutions to product related issues. For unlisted issues please contact your local GE Healthcare service representative.

General advice to achieve good performance

Before using the system make sure that:

Correct system has been selected in UNICORN System Control
Correct wavelength has been set for UV/UPC monitor
All tubing has been properly connected
All connectors are free from leakage
No tubing is folded or twisted
Online filter, if used, is changed on a regular basis
Correct buffers are used for the chosen columns and proteins
All inlet tubing has been immersed in correct buffer solutions
Enough buffer has been prepared
Buffers have been equilibrated to the environment temperature
Buffers/eluents have been degassed if necessary (e.g., in RPC runs)
Suitable columns have been selected for the target proteins
Columns have been cleaned and prepared according to column instructions
Samples have been clarified by centrifugation and/or filtration prior to sample loading
Samples have been adjusted to binding buffer conditions
Auto sampler (if used) has been prepared according to user manual
The fraction collector has been filled with appropriate number of microtiter plates or tubes
Appropriate arrangement for waste handling has been prepared

Fraction collector

Issues related to Fraction collector

Too many commands pending in the fraction collector. There is a risk that the tube is overfilled and fractionation movements lost.

The tray is loose in the frac

The internal temperature of the fraction collector is too high

The fraction collector failed to detect the code on the cassette

The fraction collector dispenser head failed to detect the flow properly and has switched to tube change with reduced accumulator functionality

The fraction collector dispenser head failed to detect a drop and has switched to tube change without Drop sync

The fraction collector dispenser head failed to detect a drop

The fraction collector cannot be found by the instrument

The accumulator is jammed or there is an internal error in the instrument

Fraction collector arm is blocked or internal fault in the fraction collector

Calibration of the drop sync sensor failed

Inlet valves

Issues related to inlet valves

The inlet valves include Inlet valve A, Inlet valve B and sample inlet valve.

The valve is not switching or is switching to wrong position

The internal valve temperature is too high

The inlet valve is not found by the instrument

Internal leakage

Faulty air sensor in the valve

External leakage

Conductivity monitor

Issues related to Conductivity monitor

Baseline drift of noisy signal

Possible cause	Suggested Remedy
Air in the conductivity flow cell.	Use the flow restrictor. Remove the air by flushing the flow cell with water or buffer.
Leaking tubing connections	Tighten the connectors. If necessary, replace the connectors.
Dirty conductivity flow cell	Clean the conductivity flow cell. See Section 7.5.8, in the User Manual
Poor mixing function	Check the mixer chamber size and change chamber if necessary. See Section 5.4.5, in the User Manual. Check the motor operation of the mixer. Place a magnet close to the mixer chamber during run. The magnet should be moving. Check that the mixer chamber is free from solids. To replace the online filter, see Section 7.7.3, in the User Manual.

Conductivity measurement with the same buffer appears to decrease/increase over time.

Ghost peaks appear in the gradient profile

Incorrect or unstable reading

Possible cause	Suggested Remedy
Poor mixing function	Check that the correct mixer chamber size is used. See Section 5.4.5, in the User Manual, for recommendations. To change the mixer, see Section 7.7.2, in the User Manual. Check the motor operation of the mixer. Place a magnet close to the mixer chamber during run. The magnet should be moving. Check that the mixer chamber is free from solids. To open the mixer, see Section 7.7.3, in the User Manual.
The column is not equilibrated	Equilibrate the column. Use the method phase *Equilibration. If necessary, clean the column. Use the predefined method Column CIP*. See Section 7.5.3, in the User Manual.
The temperature compensation factor is not properly set	Use a temperature compensation factor. The temperature compensation factor is found in *System Control:System Settings:Conductivity*. Instruction regarding the factor is also found in Section 7.6.3, in the User Manual.

Non-linear gradients or slow response to %B changes

Temperature out of range for calibration

The cell constant measurement has been aborted

The Conductivity monitor is not found by the instrument

Unstable conductivity

Waves on the gradient

Injection valve

Issues related to injection valve

The valve is not switching or is switching to wrong position

The internal valve temperature is too high

The Injection valve is not found by the instrument

Leakage

UV monitor and UV detection unit

Issues related to UV monitor and UV detection unit

UV cell path length unreadable

Unstable signal

Too low UV lamp intensity

Possible cause	Suggested Remedy
The detector is not correctly fitted	Check that the detector is fitted correctly. See Section 7.7.5, in the User Manual If this error is recurrent, contact Service.
Dirty optical fiber	Clean the connectors. See Section 7.7.5, in the User Manual.
Dirt on optical sensors in detector	Remove visible dirt on detector photo diodes. Use lint free lens paper winded around a thin wood splinter (e.g., a match or toothpick). Clean the sensors with Isopropanol through the small hole of the metal plate which covers the sensors. Dry the sensors with clean and dry lint free lens paper.
Worn-out or broken lamp	Contact Service.

The UV module is not found by the instrument

The internal temperature of the UV monitor is too high

No light transmission through the UV cell

Ghost peaks

Possible cause	Suggested Remedy
Air in the UV flow cell	Use the flow restrictor. Use the pH valve instruction to manually activate the flow restrictor (Flow path:pH valve:Restrictor in-line), or select the flow restrictor in the Method Settings phase of a method. Remove the air by flushing the cell with water or buffer. If persistent, clean the UV cell. See Section 7.4.1, in the User Manual.
Air in buffers	De-gas if necessary
Dirt in the UV flow cell	Clean the UV cell. See Section 7.4.1, in the User Manual.
Dirt in the flow path	Clean the system in accordance to Section 7.5.2, in the User Manual. Clean the column in accordance to Section 7.5.3, in the User Manul.

Baseline drift or noisy signal

Possible cause	Suggested Remedy
Flow restrictor in off-line position	Use the flow restrictor. Use the pH valve instruction to manually activate the flow restrictor (Flow path:pH valve:Restrictor in-line), or select the flow restrictor in the Method Settings phase of a method.
Air in the UV flow cell	Use the flow restrictor. Remove the air by flushing the cell with water or buffer. If persistent, clean the UV cell. See Section 7.4.1, in the User Manual.
Air in buffers	De-gas if necessary.
Impure buffers	Check if the signal is noisy with water.
Dirty optical fiber connectors	Clean the connectors. See Section 7.7.5, in the User Manual.
Dirt in the UV flow cell	Perform a System CIP. See Section 7.5.2, in the User Manual. If necessary, manually clean the UV cell. See Section 7.4.1, in the User Manual.

Autozero out of accepted range

pH monitor and pH valve

Issues related to pH monitor and pH valve

Unstable pH signal

The pH module is not found by the instrument

The internal valve temperature is too high

Temperature reading error

It is not possible to inject calibration solution

Drift of pH signal when the pH electrode has been removed from storage solution

Dummy modules

Issues related to Dummy modules

The instrument is unable to find some of the modules

Quarternary valve

Issues related to Quarternary valve

Wrong mixing ratio for Q-valve

Unexpected values for e.g., pH curve or conductivity curve using a quaternary flow

The valve is not switching or is switching to wrong position

The internal valve temperature is too high

The inlet valve is not found by the instrument

Internal leakage

External leakage

Outlet valve

Issues related to Outlet valve

The internal valve temperature is too high

The Outlet valve is not found by the instrument

Column valve

Issues related to Column valve

The Column valve is not found by the instrument

The internal valve temperature is too high

Pressure monitors

Issues related to Pressure monitors

The pressure monitors are not found by the instrument

The internal temperature of the pressure monitor is too high

Pressure offset

Irregular pressure curves

Possible cause	Suggested Remedy
Air bubbles are passing through or are trapped in the pump.	Check that there is a sufficient volume of buffer present in the flasks. Check all connections for leaks. Check pump pressure curves. See Section 8.5, in the User Manual, for example of pump pressure curves. Purge the pumps, see Section 5.5.2, in the User Manual.
The check valve does not function correctly	Remove any solids in the valves by cleaning the check valves according to the instructions in Section 7.5.7, in the User Manual.
Piston seal is leaking	Replace the piston seal according to the instructions in Sections 7.7.9, in the User Manual.
Blockage of flow path	Use the predefined method Prepare System to flush through to clear blockage. If necessary, replace tubing. See Section 7.7.1, in the User Manual. Check the mixer online filter. It can be clogged if unfiltered buffers or samples are applied. See Section 7.7.3, in the User Manual, for instructions how to replace the mixer online filter. Check the inlet filters. They can be clogged if unfiltered buffers or samples are applied. To replace the filters, see Section 7.7.7, in the User Manual. Check the column. It can be clogged if unfiltered buffers or samples are applied. To clean a column, see Section 7.5.3, in the User Manual.

Excessively high pressure values

System	Height (mm)	Footprint (mm x mm)	Weight (kg)	Flow rate (ml/min)	Pressure limit (MPa)
ÄKTA avant 25	660	860 x 710	116	0.001-25	20
ÄKTA avant 150	660	860 x 710	116	0.001-150 (normal range) 0.001-300 (column packing flow)	5
ÄKTAexplorer 10	620	500 x 460	75	0.001-10	25
ÄKTAexplorer 100	620	500 x 460	75	0.01-100	10
ÄKTAFPLC	470	380 x 480	50	0.05-20	5
ÄKTAmicro	610	480 x 450	55	0.001-2	35
ÄKTApilot	900	750 x 540	114	4-400 (full gradients) 4-800 (limited gradients)	2
ÄKTAprime plus	530	400 x 450	13	0.1-50	1
ÄKTApurifier 10	620	500 x 460	75	0.001-10	25
ÄKTApurifier 100	620	500 x 460	75	0.01-100	10
ÄKTAxpress	660	490 x 250	30	0.1-65	3
Ettan LC	610	480 x 450	55	0.001-2	35
Ettan MDLC	710	700 x 640	105	0.001-2	35
Ettan microLC	1150	650 x 500	77	0.001-2	35
Ettan nanoLC	1150	650 x 500	77	0.001-2	35

System	Height (mm)	Footprint (mm x mm)	Weight (kg)	Flow rate (ml/min)	Pressure limit (MPa)
ÄKTA avant 25	660	860 x 710	116	0.001-25	20
ÄKTA avant 150	660	860 x 710	116	0.001-150 (normal range) 0.001-300 (column packing flow)	5
ÄKTAexplorer 10	620	500 x 460	75	0.001-10	25
ÄKTAexplorer 100	620	500 x 460	75	0.01-100	10
ÄKTAFPLC	470	380 x 480	50	0.05-20	5
ÄKTAmicro	610	480 x 450	55	0.001-2	35
ÄKTApilot	900	750 x 540	114	4-400 (full gradients) 4-800 (limited gradients)	2
ÄKTAprime plus	530	400 x 450	13	0.1-50	1
ÄKTApurifier 10	620	500 x 460	75	0.001-10	25
ÄKTApurifier 100	620	500 x 460	75	0.01-100	10
ÄKTAxpress	660	490 x 250	30	0.1-65	3
Ettan LC	610	480 x 450	55	0.001-2	35
Ettan MDLC	710	700 x 640	105	0.001-2	35
Ettan microLC	1150	650 x 500	77	0.001-2	35
Ettan nanoLC	1150	650 x 500	77	0.001-2	35

Maintenance	See action
Calibrate the pH monitor	Section 7.7.1 Calibrate the pH monitor.

Maintenance	See action
Calibrate pressure monitors	Section 7.7.2 Calibrate the pressure monitors.
Change pump rinsing solution	Section 7.3.1 Change pump rinsing solution.

Maintenance	See action
Check the flow restrictor	Section 7.4.1 Check the flow restrictor.

Maintenance	See action
Clean the UV flow cell	Section 7.5.1 Clean the UV flow cell.
Replace pH electrode	Section 7.5.2 Replace the pH electrode.

Maintenance	See action
Clean the instrument externally	Section 7.6.1 Clean the instrument externally.
Perform System CIP	Section 7.6.2 Perform System CIP.
Perform Column CIP	Section 7.6.3 Perform Column CIP.
Clean the Fraction collector	Section 7.6.4 Clean the Fraction collector.
Replace tubing and connectors	Section 7.8.1 Replace tubing and connectors.
Storage of pH electrode	Section 7.6.5 Storage of the pH electrode.
Clean the pH electrode	Section 7.6.6 Clean the pH electrode.
Clean the conductivity flow cell	Section 7.6.8 Clean the conductivity flow cell.
Calibrate the Conductivity monitor	Section 7.7.3 Calibrate the Conductivity monitor.
Calibrate the UV monitor	Section 7.7.4 Calibrate the UV monitor.
Replace mixer	Section 7.8.2 Replace the mixer.
Replace online filter	Section 7.3.2 Replace the online filter.
Replace o-ring in mixer	Section 7.8.3 Replace the o-ring of the mixer.
Replace UV flow cell	Section 7.8.4 Replace the UV flow cell.
Replace flow restrictor	Section 7.8.5 Replace the flow restrictor.
Replace inlet filters	Section 7.8.6 Replace inlet filters.
Clean the check valves	Section 7.6.7 Clean the pump head check valves.
Replace check valves	Section 7.8.7 Replace the pump head check valves.
Replace pump piston seals	Section 7.8.8 Replace pump piston seals of Pump P9 or P9H. and Section 7.8.9 Replace pump piston seals of pump P9-S.
Replace pump pistons	Section 7.8.10 Replace pump pistons.
Replace pump rinsing system tubing	Section 7.8.11 Replace pump rinsing system tubing.
Replace valve modules	Section 7.8.12 Replace a valve module.

Possible cause	Suggested Remedy
Too many commands are pending in the fraction collector	The reason could be that too many fraction collector instructions have been sent. Wait for a while and try again.

Possible cause	Suggested Remedy
The cassette tray positioning discs in the frac chamber are missing	Replace the cassette tray positioning discs in the frac chamber. See Section 7.5.4, in the User Manual.

Possible cause	Suggested Remedy
Hot surroundings	Decrease the room temperature. Maximum operating temperature is 35°C.
Hardware error	Switch off the instrument and wait until the temperature has decreased. Restart the instrument. If this error is recurrent, generate a System error report and contact Service.

Possible cause	Suggested Remedy
Dirty cassette type code reader	Clean the dispenser head and its four diode windows using a cloth and a mild cleaning agent or 20% ethanol. See Section 7.5.4, in the User Manual, for more information.
Dirty cassette type codes	Clean the cassette type codes. See Section 7.5.4, in the User Manual, for more information. If this error is recurrent, set the fraction collector configuration manually in UNICORN. In System Control, select System:Settings. Navigate to Fraction collector:Cassette configuration and select Manual.

Possible cause	Suggested Remedy
Too high flow rate	Decrease the flow rate.
Air in the flow path	Check the flow path for air. Fill system and purge pumps according to Section 5.5, in the User Manual. If this error is recurrent, generate a System error report and contact Service.
Dirty drop sync sensor diode windows	Clean the drop sync sensor diode windows. See Section 7.5.4, in the User Manual. If this error is recurrent, generate a System error report and contact Service.

Possible cause	Suggested Remedy
The cable between the fraction collector and the ICU is not connected	Generate a System error report and contact Service.
A fuse in the instrument ICU is broken	The ICU needs to be changed. Generate a System error report and contact Service.

Possible cause	Suggested Remedy
Salt crystals or protein residuals block the accumulator.	Restart the instrument and perform an accumulator wash.
Mechanical error	If this error is recurrent, generate a System error report and contact Service.

Possible cause	Suggested Remedy
Obstruction inside the fraction collector.	Switch off the instrument and check for obstruction inside the fraction collector. Try to move the Frac arm by hand. Switch on the instrument. If this error is recurrent, generate a System error report and contact Service.

Possible cause	Suggested Remedy
The air intake is covered.	Uncover the air intake.
Hot surroundings	Decrease the room temperature. Maximum operating temperature is 35°C.
Hardware error	Switch off the instrument and wait until the temperature has decreased. Restart the instrument. If this error is recurrent, generate a System error report and contact Service.

Possible cause	Suggested Remedy
Wrong Node ID	Check the module´s Node ID. If necessary, change the Node ID. See Section 9.3.4, in the User Manual.
The cable between the inlet valve and the ICU is not connected	Remove the valve and make sure that the cable is connected. See Section 9.3.1, in the User Manual.

Possible cause	Suggested Remedy
Broken valve.	Replace the valve. See Section 11, in the User Manual, for ordering information and Section 9.3.2, in the User Manual, for installation instructions.

Possible cause	Suggested Remedy
Leaking tubing connectors.	Check the tubing connectors. Tighten or replace the connectors. To order connectors, see Ordering information in Chapter 11, in the User Manual.

Possible cause	Suggested Remedy
The conductivity monitor needs to be calibrated	Check calibration with a solution with known conductivity. Calibrate the conductivity monitor. See Section 7.6.3, in the User Manual.
The ambient temperature may have decreased/increased	Use a temperature compensation factor. The temperature compensation factor is found in *System Control:System Settings:Conductivity*. Instruction regarding the factor is also found in Section 7.6.3, in the User Manual.
Dirty conductivity flow cell	Clean the conductivity cell. See Section 7.5.8, in the User Manual.

Possible cause	Suggested Remedy
Air bubbles are passing through the flow cell	Check for loose tubing connections. Use the flow restrictor.
A charged particle has been detected	Prepare the sample so that charged particles are eliminated.

Possible cause	Suggested Remedy
The mixer chamber is too large	Change to a mixer chamber with a smaller volume. See Section 5.4.5, in the User Manual, for recommendations. To change the mixer, see Section 7.7.2, in the User Manual.
Bad pump function	Make sure that the pump operates properly. See Section 8.5, in the User Manual for example of pump pressure curves.
Make sure that the tubing has been washed properly.	Dirt in the tubing

Possible cause	Suggested Remedy
This error could only occur when the temperature compensation is turned on. The error will occur when the temperature is outside the range 2°C to 40°C.	Make sure the temperature of the calibration solution is within 2°C and 40°C.

Possible cause	Suggested Remedy
Internal errors	See error log. Restart instrument and retry. If this problem recurs, generate a System error report and contact Service.

Possible cause	Suggested Remedy
The conductivity monitor cable has come loose, for example when turning the instrument	Switch off the instrument. Reconnect the conductivity monitor cable (the black cable in the illustration below). Restart the instrument.

Possible cause	Suggested Remedy
Solids in the conductivity flow cell	Clean the conductivity cell. See Section 7.5.8, in the User Manual.
Air in the conductivity flow cell	Flush the conductivity flow cell with water.

Possible cause	Suggested Remedy
The cable between the Injection valve and the ICU is not connected.	Remove the valve and make sure that the cable is connected. See Section 9.3.1, in the User Manual.
Wrong Node ID.	Check the module´s Node ID. If necessary, change the Node ID. See Section 9.3.4, in the User Manual.

Possible cause	Suggested Remedy
Leaking connectors.	Check the tubing connectors. Tighten or replace the connectors. Make sure to have a syringe or a stop plug in the injection port.

Possible cause	Suggested Remedy
No UV flow cell is attached	Attach UV cell. See Section 7.7.5, in the User Manual.
UV flow cell is not correctly installed.	Please verify that the UV cell is correctly installed. See Section 7.7.5, in the User Manual.
The UV flow cell is broken	Replace the cell. See Section 7.7.5, in the User Manual.

Possible cause	Suggested Remedy
Communication problem	Contact Service

Possible cause	Suggested Remedy
The air intake is covered	Uncover the air intake.
Hot surroundings	Decrease the room temperature. Maximum operating temperature is 35°C.
Hardware error	Switch off the instrument and wait until the temperature has decreased. Restart the instrument. If this error is recurrent, generate a System error report and contact Service.

Possible cause	Suggested Remedy
Wrong wavelength for current buffer	Change wavelength or buffer.
Dirt in the UV flow cell	Clean the UV cell, see Section 7.4.1, in the User Manual.
Dirty optical fiber connectors	Clean the connectors. See Section 7.7.5, in the User Manual.
Broken UV flow cell	Replace UV flow cell. See Section 7.7.5, in the User Manual.

Possible cause	Suggested Remedy
Calibration time out	Check the connections between pH electrode and pH monitor. Regenerate the pH electrode. Place the electrode in deionized water for 30 minutes followed by 30 minutes in a buffer with pH 4. If persistent, replace the pH electrode. See Section 7.4.2, in the User Manual.
Bad pH electrode	Regenerate the pH electrode. Place the electrode in deionized water for 30 minutes followed by 30 minutes in a buffer with pH 4. Clean the pH electrode. See Section 7.5.6, in the User Manual If persistent, replace the pH electrode. See Section 7.4.2, in the User Manual.
Wrong mixer size for the used flow rate	Use the recommended mixer size for the used flow rate. See Section 5.4.5, in the User Manual.

Possible cause	Suggested Remedy
The cable between the pH valve and the ICU is not connected	Remove the pH valve and make sure that the cable is connected. See Section 9.3.1, in the User Manual.

Possible cause	Suggested Remedy
The temperature compensation of the pH monitor is turned off.	Contact Service.