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ÄKTA avant Operating Instructions

1. Sample Inlet Valve Outlet Valve 36 Holder rails KTA avant Operating Instructions 29 1015 56 AA 3 System description 3 1 AKTA avant instrument overview Available modules The AKTA avant instrument is always delivered with the standard modules installed but one or more optional modules may be added to the flow path The following tables contains information on the standard modules and the optional modules of the AKTA avant 25 and AKTA avant 150 instruments The sections that follows contain descriptions of the modules Note The valves for AKTA avant 25 and AKTA avant 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 AKTA avant 25 will give too high back pressure if used above 50 ml min The larger volumes in the H valves for AKTA avant 150 may decrease resolution and increase peak broadening if used in AKTA avant 25 Standard modules Module Label in KTA avant 25 KTA avant 150 System Pump A System Pump B Sample Pump Pressure Monitor Mixer Injection Valve Quarternary Valve Inlet Valve A Inlet Valve B Sample Inlet Valve Column Valve pH Valve Outlet Valve UV Monitor Conductivity Monitor Built in fraction collector KTA avant Operating Instructions 29 1015 56 AA ar 3 System description 3 1 AKTA avant instrument
2. GE and GE monogram are trademarks of General Electric Company AKTA is a trademark of General Electric Company or one of its subsidiaries BD is a trademark of Becton Dickinson and Company Decon is a trademark of Decon Laboratories Limited Eppendorf is a trademark of Eppendorf AG Microsoft and Windows are registered trademarks of Microsoft Corporation NUNC and Thermo Scientific are trademarks of Thermo Fisher Scientific or its subsidiaries SARSTEDT is a trademark of SARSTEDT AG amp CO Seahorse Bioscience is a trademark of Seahorse Bioscience Inc Triton is a trademark of Union Carbide Chemicals and Plastic Company Inc Tween is a trademark of Unigema Americas LLC VWR is a trademark of VWR International LLC All other third party trademarks are the property of their respective owner Any use of UNICORN is subject to GE Healthcare Standard Software End User License Agreement for Life Sciences Software Products A copy of this Standard Software End User License Agreement is available on request 2015 General Electric Company All rights reserved First published Mar 2015 All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them A copy of these terms and conditions is available on request Contact your local GE Healthcare repre sentative for the most current information GE Healthcare Europe GmbH Munzinger Strasse 5 D 79111 Freiburg German
3. Laboratory equipment The equipment listed in the following table is required during the installation procedure and should be provided at the installation site Equipment Specification Flasks liquid containers For buffers and waste Gloves For protection Protective glasses For protection KTA avant Operating Instructions 29 1015 56 AA 61 4 Installation 4 1 Site preparation 4 1 6 Required materials Fraction collector tubes The tubes used in the built in fraction collector must fulfill the requirements listed in the following table Examples of manufacturers are also listed in the table Tube size Diameter mm Height mm Max Examples of ml volume manufacturers Min Max Min Max ml NUNC NUNC SARSTEDT Thermo Scientific BD Biosciences VWR BD Biosciences BD Biosciences Deep well plates Requirements The deep well plates used in the built in fraction collector must fulfill the requirements listed in the table below Property Specification No of wells 24 48 or 96 Shape of wells Square not cylindrical Well volume 10 5 or 2 ml 62 AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 1 Site preparation 4 1 6 Required materials Approved deep well plates The plates listed in the table below are tested and approved by GE to be used with the built in fraction collector Plate type Manufacturer
4. permanently end the run click the End button Note When ending a method run in advance it is possible to save the partial result AKTA avant Operating Instructions 29 1015 56 AA 129 6 Runamethod 6 2 Applying the sample 6 2 Applying the sample Introduction A number of different sample application techniques are available The sample can be applied either directly onto the column using the sample pump or via a loop A loop can be filled either manually or by use of the sample pump This section describes sample application using a syringe to manually fill a sample loop The two stages of the sample application are described in the following table For detailed instructions and information regarding the different sample applications techniques see AKTA avant User Manual Stage Description Load The sample loop is filled with sample Inject The sample is injected onto the column How to fill a sample loop Follow the instructions to fill the sample loop with sample Step Action 1 Connect a suitable sample loop to the Injection Valve ports LoopF fill and LoopE empty 2 Fill a syringe with sample AKTA avant Operating Instructions 29 1015 56 AA 6 Runa method 6 2 Applying the sample Step Action 3 Connect the syringe to the Injection Valve port Syr 4 Load sample into the sample loop To avoid sample loss due to siphoning leave the syringe in the port until the sample has been injec
5. 7 Close the drawer Make sure that it snaps into closed position Result After the door has been closed the fraction collector arm scans the cassette type code of each cassette to identify the cassette types If deep well plates are used the instrument also identifies the types of deep well plates Rack for 50 ml tubes and rack for 250 ml bottles Step Action 1 Place 50 ml tubes or 250 ml bottles in the corresponding rack 2 Open the fraction collector drawer by pressing the handle upwards and pulling out the drawer 122 AKTA avant Operating Instructions 29 1015 56 AA 5 Prepare the system for a run 5 7 Prepare the built in fraction collector Step Action 3 Place the rack on the tray support of the fraction collector drawer Make sure that the front of the rack marked with the GE monogram faces the front of the drawer and is hooked onto the two pins a fus Ah MF ri Ci ye y ral Note The cassette tray shall not be used when the rack for 50 ml tubes or the rack for 250 ml bottles is placed in the fractionation collector drawer 4 Close the drawer Make sure that it snaps into closed position Cassette and tray identification After the fraction collector drawer has been closed the fraction collector arm scans the cassette type code of each cassette or tray to identify the cassette types If deep well plates are used the instrument also identifies the types of deep we
6. Calibrate the conductivity monitor Calibrate the UV monitor Calibrate pressure monitors Replace the mixer Replace O ring in mixer Replace the UV flow cell Replace the flow restrictor Replace inlet filters Wipe off excess oil from pump heads Clean the check valves See Section 7 5 Clean pump head check valves on page 170 Replace check valves Replace pump piston seal O rings and rinse membrane See Sec tion 7 2 Replace pump piston seal O rings and rinse membrane of Pump P9 or Pump P9H on page 149 and Section 7 3 Replace pump piston seal O ring and rinse membrane of Pump P9 S on page 160 Replace pump pistons See Section 7 4 Replace pump pistons on page 168 KTA avant Operating Instructions 29 1015 56 AA 147 7 Maintenance 7 1 Maintenance program Maintenance action Replace pump rinsing system tubing Replace valve modules 148 KTA avant Operating Instructions 29 1015 56 AA 7 Maintenance 7 2 Replace pump piston seal O rings and rinse membrane of Pump P9 or Pump P9H 7 2 Replace pump piston seal O rings and rinse mem brane of Pump P9 or Pump P9H Introduction Follow the instructions to replace the O rings piston seal and rinse membrane of pumps P9 POH A P9H B or P9H S Note Always replace the O rings piston seals and rinse membranes of both pump heads of a pump at the same time Tip A sign of internal leakage is that the pump rin
7. tion menu Click Insert Select Pumps and Pressures Sample flow and set the Flow rate to 25 0 ml min Click Insert Click Execute Result A sample flow starts KTA avant Operating Instructions 29 1015 56 AA 7 Maintenance 7 2 Replace pump piston seal O rings and rinse membrane of Pump P9 or Pump P9H Step Action 6 Run the flow for 2 hours 7 Discard the used buffer AKTA avant Operating Instructions 29 1015 56 AA 159 7 Maintenance 7 3 Replace pump piston seal O ring and rinse membrane of Pump P9 S 7 5 Replace pump piston seal O ring and rinse mem brane of Pump P9 S Introduction Follow the instructions to replace the O ring piston seal and rinse membrane of Pump P9 S Note Always replace the O rings piston seals and rinse membranes of both pump heads of a pump at the same time NOTICE e Donot disassemble the pump head unless there is good reason to believe that there is an internal leakage A sign of leakage is increasing pump rinsing solution volume Always make sure that sufficient spare components are available before attempt ing to replace a spare part e Replacing spare parts Read the instructions carefully For example some individual parts of the pump head can be as sembled incorrectly Check the orientation of each part before continuing with the next instruction Maintenance interval Replace the O ring piston seal and rinse membrane of Pump P9 S if they are d
8. 186 purge system pumps 83 replace pump pistons 168 replace pump piston seals of Pump P9 or P9H 149 replace pump piston seals of Pump P9 S 160 Purpose of this manual 5 R Recycling disposal 30 Recycling procedures 30 Reference information chemical resistance guide 176 system specifications 174 Regulatory compliance 11 Regulatory information 8 international standards 9 Room requirements introduction 52 Run after run procedures 142 monitor a run 139 start 133 S Safety notices 7 Safety precautions 15 emergency procedures 27 general precautions 15 installing and moving the instrument 18 50 labels 24 system operation 54 Software overview 44 software modules 45 Storage column 143 pH electrode 143 system 143 System Control module description 46 icons 47 process picture 140 System label 25 System preparation before preparation 102 System recommendations computer specifications 59 System storage 143 T Technical specifications system specifications 174 Typographical conventions 5 U UNICORN 44 connect to system 81 Log on 79 Start 79 System Control module 46 User information important 6 W Waste tubing prepare 71 108 KTA avant Operating Instructions 29 1015 56 AA For local office contact information visit www gelifesciences com contact GE Healthcare Bio Sciences AB Bj rkgatan 30 751 84 Uppsala Sweden www gelifesciences com avant
9. Mixer M9 Mixes the buffers delivered from the system pumps to a homogeneous buffer composition Three mixer chambers are available for AKTA avant 25 Available volumes are 0 6 ml 1 4 ml mounted at delivery and 5 ml Three mixer chambers are available for AKTA avant 150 Available volumes are 1 4 ml 5 ml mounted at delivery and 15 ml CAUTION Risk of explosion Do not use Mixer chamber 15 ml with an AKTA avant 25 sys tem configuration The maximum pressure for Mix er chamber 15 ml is 5 MPa 50 bar Inlet Valve A V9 IA or V9H IA Inlet valve for System Pump A with seven inlet ports and integrated air sensor Inlet Valve B V9 IB or V9H IB Inlet valve for System Pump B with seven inlet ports and integrated air sensor Sample Inlet Valve V9 IS or Inlet valve for sample solution with eight inlet V9H IS ports seven sample inlets and one buffer inlet and integrated air sensor AKTA avant Operating Instructions 29 1015 56 AA 39 3 System description 3 1 AKTA avant instrument overview Module Description Injection Valve V9 Inj or Valve which directs sample onto the column V9H Inj Column Valve V9 C or V9H C Column valve which connects up to five columns to the instrument and directs the flow to one column ata time The column valve features two integrated pressure sensors Allows the user to choose flow direction through the column or to bypass the column pH Valve
10. e g 0 2 ml min e Click Set flow rate J 0 200 mimin Conc B E 0 0 B Set B 100 Dy IFFEFE 1ararnh A a Result A system flow of 0 2 ml min starts 6 When buffer leaves the tubing on port 1A if port 1A was chosen in the method to be run in a continuous mode and the top part of the column is filled with buffer connect the tubing to the top of the column AKTA avant Operating Instructions 29 1015 56 AA 113 5 Prepare the system for a run 5 4 Connect a column Step Action 7 Connect a piece of tubing to the bottom of the column 8 When buffer leaves the tubing at the bottom of the column in a continuous mode connect this piece of tubing to the column valve Use the port opposite to the one already connected to the column in this example port 2B 9 Click the End button in the System Control toolbar to end the run 114 AKTA avant Operating Instructions 29 1015 56 AA 5 Prepare the system for a run 5 5 Set pressure alarms 5 5 Set pressure alarms Introduction The columns can be protected by two different types of pressure alarms e The precolumn pressure alarm protects the column hardware e The delta column pressure alarm protects the column media Column Valve label V9 C and V9H C have built in pressure sensors that automatically measure the precolumn and delta column pressure See the instructions in the next topic to set the pressure alarm for the column to be used in the run
11. resulting in liquid spillage and shat tered glass Squeezed tubing may cause increase in pressure or block liquid flow To avoid the risk of knocking over bottles always place bottles on the buffer tray and close the doors before rotating the instrument e Fasten bottles and cassettes Always fasten bottles and cas settes to the rails at the front and side panel Use appropriate holders for bottles Shattered glass from falling bottles may cause injury Spilled liquid may cause fire hazard and personal injury e Electrical shock hazard after spillage If there is a risk that large volumes of spilled liquid may penetrate the casing of the instrument immediately switch off the instrument disconnect the power cord and contact an authorized service engineer e Moving parts in fraction collector Do not open the built in fraction collector door when the instrument is running e Using a Superloop After loading a Superloop always plug the Syr port on the injection valve with a stop plug With a Super loop connected to the valve an over pressure may be created during injection e Over pressure Never block the outlet tubing with for instance stop plugs since this will create over pressure and might result in injury e 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 mainte nance 20 AKTA avant Opera
12. 50 C to 60 C Hz Power consumption 800 VA Enclosure protective class IP 21 wet side IP 22 Tubing and connectors Inlet FEP tubing inner diameter i d 1 6 mm Tubing connector 5 16 Ferrule yellow 1 8 Pump to Injection valve PEEK tubing i d 0 75 mm Fingertight connector 1 16 After Injection valve PEEK tubing i d 0 50 mm Fingertight connector 1 16 Outlet and waste ETFE tubing i d 1 0 mm Fingertight connector 1 16 Environmental ranges Parameter Storage and transport temperature range 25 C to 60 C Chemical environment See Section 8 2 Chemical resistance guide on page 176 174 AKTA avant Operating Instructions 29 1015 56 AA Operating range Parameter Operating temperature range 8 Reference Information 8 1 System specifications 4 C to 35 C Relative humidity Equipment noise level Equipment AKTA avant instrument 20 to 95 noncondensing Acoustic noise level KTA avant Operating Instructions 29 1015 56 AA lt 70 dBA 175 8 Reference Information 8 2 Chemical resistance guide Oz Introduction Chemical resistance guide This section specifies the chemical resistance of the KTA avant instrument to some of the most commonly used chemicals in liquid chromatography Biocompatibility The KTA avant instrument is designed for maximum biocompatibility with biochemically inert flow paths constructed mainly from t
13. 6 202 859 9 Dithioerythritol DTE 3483 12 3 222 468 7 Dithiothreitol DTT 3483 12 3 222 468 7 Ethylenediaminetetraacetic acid EDTA 60 00 4 200 449 4 Mercaptoethanol 37482 11 4 253 523 3 Potassium chloride Other substances Chemical Acetone Concentra tion 7447 40 7 231 211 8 CAS no EC no 67 64 1 200 662 2 Ammonia Dimethyl sulphoxide DMSO 7664 41 7 231 635 3 67 68 5 200 664 3 Ethanol for long term stor age Phosphoric acid 75 08 1 200 837 3 7664 38 2 231 633 2 AKTA avant Operating Instructions 29 1015 56 AA 8 Reference Information 8 3 Check and change the Node ID of a module 8 3 Check and change the Node ID of a module Introduction Node ID is a unit number designation that is used by the instrument to distinguish be tween several units of the same type All standard valves and available optional modules are pre configured to the default function However the function of a valve or module can be changed by changing the Node ID Also in a troubleshooting situation it may be useful to check a valve s or module s Node ID Note The function of a valve or module is defined by its Node ID not by its physical position Node ID for standard modules The following table lists the Node ID for the standard modules Module System Pump A P9 A or P9H A Node ID System Pump B P9 B or P9H B Sample Pump
14. AKTA avant Operating Instructions 29 1015 56 AA 43 3 System description 3 2 UNICORN software 3 2 1 UNICORN software overview 3 2 1 UNICORN software overview Introduction This section gives a brief overview of the UNICORN software a complete package for control supervision and evaluation of chromatography instruments and purification runs From hereon UNICORN refers to compatible versions of the software The examples given in this manual are from UNICORN 6 4 44 KTA avant Operating Instructions 29 1015 56 AA 3 System description 3 2 UNICORN software 3 2 1 UNICORN software overview 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 Mlole UI Main functions Administration Perform user and system setup system log and database administration Method Editor Create and edit methods using one or a combination of e Predefined methods with built in application support e Drag and drop function to build methods with relevant steps e 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 Evaluation Open results evaluate runs an
15. Al A pe aN Col iS k PS M Saye rE B1 com B Sample flow 03 A 1 000 mi min PEEP 0 50 mum Buffer HEE S PUD wheats Buffer Start pump wash e Click Set flow rate Result The sample pump flow starts 6 Connect a 25 to 30 ml syringe to the left purge valve of the sample pump Make sure that the syringe fits tightly into the purge connector 7 Open the purge valve by turning it counter clockwise about three quarters of a turn Draw 5 to 10 ml of liquid slowly into the syringe with a rate of about 1 ml s 8 Close the purge valve by turning it clockwise Disconnect the syringe and discard its contents AKTA avant Operating Instructions 29 1015 56 AA 93 4 Installation 4 5 Prime inlets and purge pump heads 4 5 2 Prime sample inlets and purge Sample Pump Step Action 9 Connect the syringe to the right purge valve on the sample pump and repeat step 6 to 8 10 Check that there is no air left in the pump by following the instructions in Validate prime or purge of System Pump A or B or Sample Pump on page 89 End the run Click the End button in the System Control toolbar to end the run El 94 KTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 5 Prime inlets and purge pump heads 4 5 3 Prime Q inlets 4 5 3 Prime Q inlets Overview The procedure consists of the following stages Stage 1 2 Description Prime all Q inlet tubing Validat
16. C E System 2 In the Connect to Systems dialog box e Select a system check box e Click Control for that system e Click OK Result The selected instrument can now be controlled by the software Tip If UNICORN ts unable to connect to the selected instrument see Chapter Troubleshooting in AKTA avant User Manual AKTA avant Operating Instructions 29 1015 56 AA 81 4 Installation 4 5 Prime inlets and purge pump heads 45 Prime inlets and purge pump heads About this section Before using the sample pump or system pumps it is important to do the following e Prime the inlets fill the inlets with liquid e Purge the pumps remove air from the pump heads This section describes how to prime the buffer inlets sample inlets and Q inlets and how to purge the system pumps and the sample pump In this section This section contains the following subsections Section See page 4 5 1 Prime buffer inlets and purge system pumps 83 4 5 2 Prime sample inlets and purge Sample Pump 90 4 5 3 Prime Q inlets 95 82 AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 5 Prime inlets and purge pump heads 4 5 1 Prime buffer inlets and purge system pumps 4 5 1 Prime buffer inlets and purge system pumps Overview The procedure consists of the following stages Stage 1 2 Description Prime all inlet tubing to be used during the run Validate priming of inlet tubing Purge System Pump B if press
17. C2 or V9H C2 Unused precolumn pressure monitor in N A 4 second Column Valve 182 AKTA avant Operating Instructions 29 1015 56 AA 8 Reference Information 8 3 Check and change the Node ID of a module Module Node ID Unused post column pressure monitor 5 in second Column Valve Second Outlet Valve V9 O2 or V9H 02 Third Outlet Valve V9 03 or V9H 03 External Air Sensor L9 1 2 or L9 1 5 I O box E9 Second O box E9 Second UV Monitor Second Conductivity Monitor Second Fraction Collector KTA avant Operating Instructions 29 1015 56 AA 183 8 Reference Information 8 3 Check and change the Node ID of a module Check and change the Node ID The Node ID of a module is set by the positions of an arrow of two rotating switches at the back of the module Follow the instructions to check or change the Node ID Step Action 1 If applicable remove the module according to the instruction in KTA avant User Manual 2 The Node ID is set by the positions of an arrow of two rotating switches at the back of the module e The first rotating switch labeled A sets the tens e The second switch labeled B sets the units e For example for Node ID 13 the A switch is set to 1 and the B switch to 3 Check the Node ID and compare it with the listed Node IDs in the tables above 4 To change the Node ID use a screwdriver to set the arrows of the switches to the desired number 5 Re
18. Instructions 29 1015 56 AA 4 Installation 4 2 Hardware installation 4 2 4 Install the Barcode Scanner 2 D and the pH electrode 4 2 4 Install the Barcode Scanner 2 D and the pH electrode Introduction This section describes how to install the Barcode Scanner 2 D and the pH electrode Install the barcode scanner Connect the cable of the Barcode Scanner 2 D to the scanner head and to a USB port on the computer Install the pH electrode If pH monitoring is to be used you need to replace the dummy electrode mounted at delivery with a pH electrode CAUTION pH electrode Handle the pH electrode with care The glass tip may break and cause injury Follow the instructions to install the pH electrode Step Action 1 Unpack the pH electrode Make sure that the electrode is not broken or dry 2 Unscrew the dummy electrode from the flow cell 3 Pull off the plug from the connector on the front of the pH valve and store the plug together with the dummy electrode 4 Remove the cover from the tip of the pH electrode 5 Carefully insert the electrode in the flow cell Tighten the locking ring by hand to secure the electrode 6 Connect the pH electrode cable to the connector on front of the pH valve KTA avant Operating Instructions 29 1015 56 AA 73 4 Installation 4 2 Hardware installation 4 2 5 Prepare the pump rinsing system 4 2 5 Prepare the pump rinsing system Illustration of the pump piston ri
19. Manual Run 0 862 ml 1 000 ml min 0 000 ml min 0 22 MPa 0 01 MPa 13 746 mAU 43 825 mAU Off amp Cond 0 00 mSiem 1 Chromatogram x i UV 1_280 Cond Conc B Fraction Injection ANN Runlog PreC pressure DeltaC pressure mali Frac temp System flow 1 000 mi min 0 0 xs Sample flow 0 000 y N Prec 0 22 mPa Cond 0 00 mS cm pH off Temp 20 2 c k Al mmm A IAS A alar oe uUVv1 13 746 mau Bsefsss Waste gt 4 a B By pass i mmm UV mm Cond mmm Restrictor mum Frac meee ton a me TA A gt a Deltac 0 01 mPa p igh alarm 20 00 set n Sse Pp 43 825 mau UV 2nd O box ManualRun Block Nowatch Connection Connected in control Controlled by Default HCE C8ZH05 Note On the View menu click Run Log to open the Run Log pane which presents all registered actions 46 AKTA avant Operating Instructions 29 1015 56 AA System Control toolbar buttons The following table shows the System Control toolbar buttons that are referred to in this manual Button Function Open Method Navigator Opens the Method Naviga tor where available meth ods are listed Button 3 System description 3 2 UNICORN software 3 2 2 The System Control module Function Run Starts a method run Hold Suspends the method run while current flow rate and valve positions are sustained Pause Suspends the method run and stops all pumps Continue Resumes f
20. P9 S or P9H S Pressure Monitor system pressure R9 Pressure Monitor sample pressure R9 Mixer M9 Injection Valve V9 Inj or V9H Inj Quaternary Valve Q9 Inlet Valve A V9 IA or V9H IA Inlet Valve B V9 IB or V9H IB Sample Inlet Valve Column Valve V9 IS or V9H IS V9 C or V9H C Precolumn pressure monitor in Column Valve Post column pressure monitor in Column Valve N A N A AKTA avant Operating Instructions 29 1015 56 AA 181 8 Reference Information 8 3 Check and change the Node ID of a module Module Nole lt H B pH Valve V9 pH or V9H pH OH Monitor H9 Note The pH monitor is included in the pH valve module box Outlet Valve V9 O or V9H O UV Monitor U9 M UV detector U9 D 0 Conductivity Monitor C9 0 Built in fraction collector Not settable by the user Node ID for optional modules The following table lists the Node ID for the optional modules Module IN fore lt H B Second Inlet Valve A V9 A2 or V9H A2 Second Inlet Valve B V9 B2 or V9H B2 Extra Inlet Valve X1 V9 IX or VOH IX Extra Inlet Valve X2 V9 IX or VOH IX Second Sample Inlet Valve V9 S2 or V9H S2 Versatile Valve V9 V or V9H V Second Versatile Valve V9 V or V9H V Third Versatile Valve V9 V or V9H V Tertiary Versatile Valve V9 V or V9H V Loop Valve V9 L or V9H L Second Column Valve V9
21. a column type was selected at method creation the Select Columns dialog box opens Continue with the steps outlined in the next topic Select Columns System3 Enter or select a column individual Code Disable column logging for this run C Apply to all methods with the same column type Methods Column Barcode Column type Column Handling Scouting run 1 HiPrep 26 10 Desalting HiPrep 26 10 Desalting HiPrep 26 10 Desalting Column Handling Scouting run 2 Column Handling Scouting run 3 e lf column logging was not chosen at installation of UNICORN and or no column type was selected at method creation the run starts directly 134 AKTA avant Operating Instructions 29 1015 56 AA 6 Runa method 6 3 Start a method run Register a column and start a run The following instructions describe how to register a column and start a run Step Action 1 Is the column to be used already registered e If No continue to step 2 e lf Yes continue to step 5 Select Columns System3 Enter or select a column individual Code Enter ID Select ID Disable column logging for this run C Apply to all methods with the same column type Methods Remark Column Barcode Column type Column Handling Scouting run 1 HiPrep 26 10 Desalting Column Handling Scouting run 2 HiPrep 26 10 Desalting Column Handling Scouting run 3 HiPrep 26 10 Desalting 2 In the Select Columns dialog box click New Re
22. acid TFA 2 TFA Ethanol 100 75 08 1 200 837 3 lsopropanol 100 67 63 0 200 661 7 Methanol 100 74 93 1 200 659 6 Water organic mobile Max 5 phase formic acid formic acid 1 Organic solvents can penetrate weaknesses in PEEK tubing walls more easily than water based buffers Special care should therefore be taken with prolonged use of organic solvents close to pressure limits Note Quaternary valve is not resistant Depending on pressure tubing between pump head and pressure monitor needs to be changed See AKTA avant User Manual for more information 2 Mobile phase system Note It is recommended to replace the mixer sealing ring with the highly resistant O ring product code 29 0113 26 if the system is to be exposed to organic solvents or high concentrations of organic acids such as acetic acid and formic acid for a longer period of time Salts and additives for hydrophobic interaction chromatography HIC Continuous use Chemical Concentra CAS no EC no tion Ammonium chloride 12125 02 9 235 186 4 Ammonium sulfate 7783 20 2 231 984 1 Ethylene glycol 107 21 1 203 473 3 Glycerol 56 81 5 200 289 5 AKTA avant Operating Instructions 29 1015 56 AA 179 8 Reference Information 8 2 Chemical resistance guide Reducing agents and other additives Continuous use Chemical Arginine Concentra tion CAS no EC no 74 79 3 200 811 1 Benzyl alcohol 100 51
23. and if applicable to set the parameters for the tubing dimensions Note Remember to lower the system pressure alarm and sample pressure alarm if the optional UV Monitor U9 L and or the optional second Conductivity Monitor C9 is used on the high pressure side in the system before the column s The UV Monitor U9 L flow cells has a maximum pressure limit of 2 MPa 20 bar and the second Conductivity Monitor C9 flow cell has a maximum pressure limit of 5 MPa 50 bar NOTICE UV and conductivity flow cells on the high pressure side When placing UV and or conductivity flow cells on the high pressure side of the column the UV flow cell has a maximum pressure limit of 2 MPa 20 bar and the conductivity flow cell has a maximum pressure limit of 5 MPa 50 bar NOTICE Remember to lower the system pressure alarm and sample pres sure alarm if the optional UV Monitor U9 L and or the optional second Conductivity Monitor C9 is used on the high pressure side in the system before the column s The UV Monitor U9 L flow cells has a maximum pressure limit of 2 0 MPa 20 bar and the second Conductivity Monitor C9 flow cell has a maximum pressure limit of 5 0 MPa 50 bar KTA avant Operating Instructions 29 1015 56 AA 115 5 Prepare the system for a run 5 5 Set pressure alarms Precolumn pressure alarms It is important that the precolumn pressure alarm is set during all runs where a column is used The pressure ala
24. and Technical Manual for more information about e licenses Step Action 1 Double click the UNICORN icon on the desktop Result The Log On dialog box opens Note If there is no connection to the database it is still possible to log on to UNICORN and control a running system The Log On dialog box will give the option to start System Control without a database Click Start System Control to proceed to the next Log On dialog box KTA avant Operating Instructions 29 1015 56 AA 79 4 Installation 4 4 Start UNICORN and connect to system Step Action 2 In the Log On dialog box e select User Name and e enter Password Note It is also possible to select the Use Windows Authentication checkbox and enter a network ID in the User Name field i Log On UNICORN E Use Windows Authentication User Name Default v Domain Access Access ToEverything v Group Start V Administration V System Control E Method Editor E Evaluation Options lt lt e click OK Result The selected UNICORN modules open 80 AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 4 Start UNICORN and connect to system Connect to system Follow the instructions to connect the instrument to UNICORN Step Action 1 In the System Control module click the Connect to Systems button System name Control View amp System E System4 E B Systems C E System
25. and you can restart the UNICORN client to regain control No data will be lost 28 AKTA avant Operating Instructions 29 1015 56 AA 2 Safety Instructions 2 3 Emergency procedures Uninterruptible power supply UPS A UPS can prevent data loss during a power failure and allow time for a controlled shut down of the AKTA avant instrument For UPS power requirements see Technical specifications on page 174 Remember to also take into account the specifications for the computer and monitor Refer to the manu facturers documentation Restart the instrument after emergency shutdown or power failure Follow the instructions to restart the instrument after an emergency shutdown or power failure Step Action 1 Make sure that the condition that caused the emergency shutdown or power failure is corrected 2 If power to the instrument has been lost restart the instrument 3 e Press the Continue button on the instrument display or e Click the Continue button in the System Control module AKTA avant Operating Instructions 29 1015 56 AA 29 2 Safety Instructions 2 4 Recycling information 2 4 Recycling information Introduction This section describes the procedures for disposal and recycling of the AKTA avant in strument Decommissioning and disposal of the equipment When taking the AKTA avant instrument out of service e The equipment must be decontaminated e The components must be separated and rec
26. bench For Pump P9 unscrew the two screws of the support washer using a star screwdriver T20 For Pump P9H unscrew the four screws of the support washer using a star screwdriver T10 Discard the O ring 1 on the support washer and the discard the piston seal 2 located in the front section of the pump head Note Be careful not to scratch the metal surfaces Pump P9H 7 Unscrew one of the two screws securing the pump membrane housing using a star screwdriver T20 Unscrew the second screw and at the same time push firmly on the front of the pump membrane housing to compensate for the pressure of the piston return spring Wily AL ey tsj vo ti KTA avant Operating Instructions 29 1015 56 AA 153 7 Maintenance 7 2 Replace pump piston seal O rings and rinse membrane of Pump P9 or Pump P9H Step Action 8 Carefully pull off the pump membrane housing together with the piston and return spring A 9 Inspect the piston and return spring for signs of damage If damaged discard the piston and return spring and use a new piston and return spring when assembling the pump head 10 Clean the pump head and pump membrane housing in an ultrasonic bath If there are particles on any surfaces the check valves should be removed and cleaned separately see Section 7 5 Clean pump head check valves on page 170 154 KTA avant Operating Instructions 29 1015 56 AA 7 Maintenance 7 2 Replace pump pi
27. buffer close to the lowest or highest pH Last calibrated on 2009 01 13 10 52 07 01 00 you need to measure as your second point Calibrated electrode slope A pH for buffer 1 Assymetry potential at pH 7 mi BH for buffer 2 v Ta PEP ET ae E eed E 2 Set the pH monitor as the monitor to calibrate by clicking pH on the Monitor to calibrate menu AKTA avant Operating Instructions 29 1015 56 AA 117 5 Prepare the system for a run 5 6 Calibrate the pH monitor Step Action 3 Click Prepare for calibration Result The pH valve switches to the calibration position 4 Enter the pH of the first pH standard buffer in the pH for buffer 1 box 5 Fill a syringe with approximately 10 ml of the first pH standard buffer Con nect the syringe to the Luer connector in pH valve port Cal and inject the buffer 6 When the Current value is stable click Calibrate 7 Wash the pH flow cell by injecting water into pH valve port Cal using a new syringe 8 Enter the pH of the second pH standard buffer in the pH for buffer 2 box 9 Repeat steps 5 to 6 using the second pH standard buffer Result The calibration date and time are displayed in the dialog box and also values for Calibrated electrode slope and Asymmetry potential at pH 7 10 Is the Calibrated electrode slope 2 80 and the Asymmetry potential at pH 7 inside the interval 60 mV e If Yes Click Close to switch the pH valve back to the default position and to
28. by the Chinese standard SJ T11363 2006 Requirements for Concentration Limits for Certain Hazardous Substances in Electronic Information Products The number in the symbol is the Environment friendly Use Period EFUP which indicates the period during which the toxic or hazardous substances or elements contained in electronic information products will not leak or mutate under normal operating con ditions so that the use of such electronic information products will not result in any severe environmental pollution any bodily injury or damage to any assets The unit of the period is Year O In order to maintain the declared EFUP the product shall be operated normally ac cording to the instructions and environmental conditions as defined in the product manual and periodic maintenance schedules specified in Product Maintenance Pro cedures shall be followed strictly Consumables or certain parts may have their own label with an EFUP value less than the product Periodic replacement of those consumables or parts to maintain the declared EFUP shall be done in accordance with the Product Maintenance Procedures This product must not be disposed of as unsorted municipal waste and must be collected separately and handled properly after decommissioning AKTA avant Operating Instructions 29 1015 56 AA 31 2 Safety Instructions 2 5 Declaration of Hazardous Substances DoHS BeA SWART ANAMRES Name and Concentration of Hazardous Substance
29. close the Calibration dialog box e If No Clean the pH electrode and repeat the calibration procedure If this does not help replace the electrode For information about cleaning and replacing the pH electrode see KTA avant User Manual Chapter Maintenance 118 KTA avant Operating Instructions 29 1015 56 AA 5 Prepare the system for a run 5 7 Prepare the built in fraction collector 5 7 Prepare the built in fraction collector Introduction This section describes how to prepare the built in raction collector For information re garding the types of deep well plates tubes and cassettes see AKTA avant User Manual WARNING Fraction collector Do not fractionate flammable liquids in the built in fraction collector When running RPC methods collect fractions through the outlet valve or the optional external Fraction Collector F9 R Prepare the fraction collector Before starting to prepare the built in fraction collector check the fractionation settings in the method to be run Perform the steps described below according to the settings in the method e Insert the cassette tray or a rack for tubes or bottles e Change the System Settings in UNICORN to set the fractionation mode and other settings for fraction collection How to insert a tray or a rack is shown in the following topic For information on how to change the System Settings before a run see UNICORN System Control Manual The available Syst
30. e Spread of biological agents The operator must take all nec essary actions to avoid spreading hazardous biological agents The facility must comply with the national code of practice for biosafety e High pressure The product operates under high pressure Wear protective glasses and other required Personal Protective Equipment PPE at all times AKTA avant Operating Instructions 29 1015 56 AA 17 2 Safety Instructions 2 1 Safety Precautions CAUTION To avoid hazardous situations when working with the AKTA avant system take the following measures for personal protec tion e Always use appropriate personal protective equipment when decommissioning the equipment e Close doors To minimize the risk of exposure to hazardous chemicals and pressurized liquids always close the foldable door and the pump cover before starting a run e Cut injuries The tubing cutter is very sharp and must be handled with care to avoid injuries Installing and moving WARNING To avoid injury when installing and moving the AKTA avant system take the following measures for personal protection e Move transport crates Make sure that the forklift has the capacity to safely lift the crate weight Make sure that the crate is properly balanced so that it will not accidentally tip when moved e Heavy object The KTA avant instrument weighs about 116 kg Use proper lifting equipment or use four or more persons when movi
31. flowpath Introduction The flow path contains tubing valves pumps and monitors This section gives an overview of the flow path and describes how to prepare the flow path before a run CAUTION e Fasten bottles and cassettes Always fasten bottles and cas settes to the rails at the front and side panel Use appropriate holders for bottles Shattered glass from falling bottles may cause injury Spilled liquid may cause fire hazard and personal injury e Max weight on buffer tray Do not place containers with a volume of more than 10 liters each on the buffer tray The total allowed weight on the buffer tray is 40 kg e Avoid spillage and overflow Make sure that the system is prepared according to the settings in the method to be run For example make sure that the waste tubing Is inserted in an appropriate waste container and secured in place 104 AKTA avant Operating Instructions 29 1015 56 AA 5 Prepare the system for a run 5 2 Prepare the flowpath Illustration of the flow path The following illustration shows an overview of the standard flow path Description Pressure Monitor Sample Pump Sample Inlet Valve Quaternary Valve Inlet Valve A Inlet Valve B 7 System Pump A AKTA avant Operating Instructions 29 1015 56 AA 105 5 Prepare the system for a run 5 2 Prepare the flowpath Description System Pump B Pressure Monitor Mixer Injection Valv
32. information is saved and the dialog box closes AKTA avant Operating Instructions 29 1015 56 AA 137 6 Runa method 6 3 Start a method run Step Action 5 In the Select Columns dialog box e Click Enter ID e Use the Barcode Scanner 2 D see step 3 to enter the column ID Select Columns System3 Enter or select a column individual Code lot exp ID Enter ID ks 9288 13 12345678 0000 00 0005 O Select ID 28 9288 13 12345678 0000 00 0005 HiPrep 261 Disable column logging for this run C Apply to all methods with the same column type Methods Column Barcode Column type Column Handling Scouting run 1 28 9288 13 12345678 0000 00 0005 HiPrep 26 10 Desalting Column Handling Scouting run 2 HiPrep 26 10 Desalting Column Handling Scouting run 3 HiPrep 26 10 Desalting e Alternatively click Select ID and click the column individual to be used in the run from the menu Select Columns System3 Enter or select a column individual Code Enter ID Select ID 28 9288 13 12345678 0000 00 0005 HiPrep 2610 Desalting E Disable column logging 28 9288 13 12345678 0000 00 0006 HiPrep Desalting 2610 E 28 9288 13 28928813 0000 00 1234 HiPrep 2610 Desalting E C Apply to all methods with the same column type Methods Remark Column Barcode Column type Column Handling Scouting run 1 28 9288 13 2892881 3 0000 00 1234 HiPrep 26 10 Desalting Column Handling Scouting run 2 HiPr
33. instrument Make sure that there is always at least 20 cm of free space around the AKTA avant instrument to allow for sufficient ventilation and rotation on the swivel foot When ro tating the instrument take care not to stretch or squeeze tubing or cables A disconnected cable may cause power interruption or network interruption Stretched tubing may cause bottles to fall resulting in liquid spillage and shattered glass Squeezed tubing may cause increase in pressure or block liquid flow To avoid the risk of knocking over bottles always place bottles on the buffer tray and close the doors before rotating the instrument Laboratory bench The bench must be clean flat and stable to support the weight of the AKTA avant system see the following table Equipment weight 54 AKTA avant Operating Instructions 29 1015 56 AA Equipment dimensions 4 Installation 4 1 Site preparation 4 1 2 Room requirements The outer dimensions of the AKTA avant instrument are shown in the following illustration 66cm Equipment weight Item KTA avant instrument Computer approximately 9 kg Monitor approximately 3 kg Total KTA avant Operating Instructions 29 1015 56 AA approximately 130 kg 55 4 Installation 4 1 Site preparation 4 1 3 Site environment 4 1 3 Site environment Introduction This section describes the environmental requirements for installation of the the AKTA avant instrume
34. membrane in the hole with 20 ethanol before mounting 3 Place the pump membrane housing onto the locating pins on the front of the pump module 4 Screw one of the two screws securing the pump membrane housing using a star screwdriver T20 Push firmly on the front of the pump membrane housing to compensate for the pressure of the piston and then screw the second screw 5 Make sure that the new seal is wetted with 20 ethanol and then tighten both screws fully 6 Reconnect the tubing of the pump piston rinsing system 7 Reconnect the inlet manifold 8 Reconnect the tubing to the pump head and reconnect the pump inlet tubing Break in the new pump piston seal Follow the instruction below to break in the new pump piston seal of Pump P9 or P9H Step Action iE Fill a buffer vessel with 20 ethanol in water Immerse the inlet tubing for example Al for System Pump A B1 for System Pump B or 1 for the Sample Pump in the buffer vessel Place the buffer vessel on the buffer tray 2 Prime the inlets and purge the pump see Section 4 5 1 Prime buffer inlets and purge system pumps on page 83 AKTA avant Operating Instructions 29 1015 56 AA 157 7 Maintenance 7 2 Replace pump piston seal O rings and rinse membrane of Pump P9 or Pump P9H Step Action 3 e For Pump P9 Connect the reference capillary Ref 2 or an equivalent capillary that gives a backpressure of 6 to 8 MPa 60 to 80 bar to one of the column positions
35. overview Optional modules Module Label in KTA avant 25 KTA avant 150 Second Inlet Valve A Second Inlet Valve B Extra Inlet Valve X1 Extra Inlet Valve X2 Second Sample Inlet Valve Versatile Valve Loop Valve Second Column Valve Second Outlet Valve Third Outlet Valve External Air Sensor L9 1 5 External Air Sensor L9 1 2 I O box Second UV Monitor Second Conductivity Monitor Second Fraction Collector Description of standard modules The following modules are installed in the instrument when delivered Module Description Quaternary Valve Q9 Valve which allows automatic mixing of four dif ferent solutions System Pump A P9 A or P9H A A high precision pump which delivers buffer in purification runs 38 KTA avant Operating Instructions 29 1015 56 AA 3 System description 3 1 AKTA avant instrument overview Module Description System Pump B P9 B or P9H B A high precision pump which delivers buffer in purification runs Sample Pump P9 S or P9H A high precision pump which delivers sample or buffer in purification runs Pressure Monitor R9 Pressure monitor which reads the system pressure after System Pump A and System Pump B Pump flow restrictor Prevents the system from siphoning if the flow path after the pump is open Gives a small back pressure to the pump in extreme low pressure applications
36. required e Adjustable wrench e 100 Methanol e Distilled water e Ultrasonic bath Instruction Follow the instructions to remove and clean the pump head check valves Step Action 1 Before taking the check valve apart always try to clean the check valves by priming the pump heads first with distilled water then with 100 Methanol and then with distilled water again 2 Switch off the instrument 3 Disconnect the tubing from the pump head and disconnect the pump inlet tubing Disconnect the tubing of the pump rinsing system 170 KTA avant Operating Instructions 29 1015 56 AA Step 7 Maintenance 7 5 Clean pump head check valves Action Unscrew the purge valve by turning it counter clockwise and lift off the metal ring Unscrew the plastic nut of the upper check valve using an adjustable wrench and gently lift off the upper check valve Q a 7 J Unscrew the two white plastic screws located below each pump head Pull the plastic connectors to the sides to release the inlet manifold T U7 a z TY o i AKTA avant Operating Instructions 29 1015 56 AA 171 7 Maintenance 7 5 Clean pump head check valves Step Action i WARNING Hazardous substances When using hazardous chemicals 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
37. update of parameters during run e Select Pumps and pressures System flow and set Flow rate to 0 01 ml min e Click Execute 3 Connect a 25 to 30 ml syringe to one of the purge valves of either of the system pumps Make sure that the syringe fits tightly into the purge connec tor 96 AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 5 Prime inlets and purge pump heads 4 5 3 Prime Q inlets Step Action 4 Open the purge valve by turning it counterclockwise about 3 quarters of a turn Draw 10 ml of liquid into the syringe Check that the Q1 inlet is filled with liquid 5 Close the purge valve by turning it clockwise Disconnect the syringe and discard its contents 6 Repeat steps 2 to 5 for Q2 Q3 and Q4 respectively by setting the respective Quaternary start concentration to 100 Tip The inlet tubing that is immersed in distilled water should be the last piece of inlet tubing to be primed Tip If you will perform a BufferPro run end with either Q1 or Q2 7 Check that there is no air left in the pump by following the instructions in Validate prime or purge of System Pump A or B or Sample Pump on page 89 If air bubbles are indicated follow the instructions in Purge Quaternary Valve and the system pumps on page 97 Purge Quaternary Valve and the system pumps If the priming was done thoroughly and the final buffer was drawn all the way into the syringe and the validation of the primi
38. 100 67 63 0 200 661 7 Sodium hydroxide 2M 1310 73 2 215 185 5 Sodium hydroxide ethanol 1 M 40 N A AKTA avant Operating Instructions 29 1015 56 AA 177 8 Reference Information 8 2 Chemical resistance guide 178 Chemical Sodium chloride Sodium hypochlorite Concentra CAS no EC no tion 4M 7647 14 5 231 598 3 10 7681 52 9 231 668 3 1 if hydrochloric acid HCl is used as a cleaning agent when columns are connected to the system the HCI concentration should not exceed 0 1 M in the pressure sensors Remember that the AKTA avant system has pressure sensors in the column valve V9 C For other parts of the system up to 1 M HCl is acceptable for short periods of use See Cleaning chemicals on page 176 Solubilization and denaturing agents Continuous use as additives in separation and purification methods Chemical Guanidinium hydrochloride Concentra CAS no EC no tion 50 01 1 200 002 3 Sodium dodecyl sulfate SDS 151 21 3 205 788 1 TRITON X 100 9002 93 1 Tween 20 9005 64 5 500 018 3 AKTA avant Operating Instructions 29 1015 56 AA 8 Reference Information 8 2 Chemical resistance guide Chemicals used in reversed phase chromatography RPC Continuous use Chemical Concentra CAS no EC no tion Acetonitrile 1 100 75 05 8 200 835 2 Acetonitrile Tetrahydrofu 85 15 109 99 9 203 726 8 1 ran Acetonitrile water Trifluo Max 0 2 N A roacetic
39. 1015 56 AA 77 4 Installation 4 3 Software installation 4 4 Software installation Introduction This section gives an overview of the different UNICORN installation types Detailed information about software installation and configuration is available in the UNICORN Administration and Technical Manual Software installations You can install UNICORN in one of the following configurations as a complete UNICORN installation on a stand alone workstation full installation as a UNICORN database and license server custom installation as a UNICORN software client and instrument server software on a network client station custom installation You can also do the following when installing UNICORN define a system as part of the installation configure E licenses configure Windows settings necessary for the UNICORN Process Picture in a network deployment configure firewall settings when necessary upgrade UNICORN remove UNICORN installations set up a system printer 78 AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 4 Start UNICORN and connect to system 44 Start UNICORN and connect to system Introduction This section describes how to start and log on to UNICORN and how to connect the in strument to UNICORN Start UNICORN and log on Follow the instructions to start UNICORN and log on to the program A valid e license must be available for the workstation See UNICORN Administration
40. 6 AA 59 4 Installation 4 1 Site preparation 4 1 5 Computer requirements e The Windows basic color scheme is recommended e Using the Windows 7 Aero color scheme is not recommended e Windows power save features should be turned off to avoid conflicts with system operations e UNICORN is not compatible with the Windows 7 feature High DPI Aware ness which allows the graphic user interface to be scaled The interface scale must remain at 100 to avoid issues with clipping and misaligning of parts of the UNICORN user interface Normally the scale is set at 100 by default 1 UNICORN must be closed when the color scheme is changed 60 AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 1 Site preparation 4 1 6 Required materials 4 1 6 Required materials Introduction This section describes the accessories required for the installation and operation of the AKTA avant instrument Buffers and solutions The buffers and solutions listed in the following table are required during the installation procedure and should be provided at the installation site Buffer solution Required volume Scope of use Distilled water 1 liter Air sensor test fraction collector test Quaternary Valve test and system test 1 acetone in distilled 0 5 liter Quaternary Valve test water 1 acetone and 1M NaC 0 5 liter System test in distilled water 20 ethanol 200 ml Priming of the pump piston rinsing system
41. 9 S ou 160 7 4 Replace OT TO SUNS aseri a cea ctee tures cites 168 7 5 Clean PUMP head CHECK VAIVE S waceecccsccssssssssseessesssesssecssecssecssecssscssecsssssssesssesssecesecssecssecssesseeseeen 170 8 Reference Information eseeseesesseseeseeseesessesseseeseesorsessesseseeseesersessesseseeseesersesses 173 oL Oyeme 1 6 0 9 srera eT Oe eT OOP ene eae 174 82 Chiemicares 2100 1 1 1 0 6 lt a nee ne nna On Ene ev ENE 176 8 3 Check and change the Node ID Of a module woe ecccessessseesseesssesssessseessecssecssecsseesseessseesseeses 181 Tie E RE eRe ore EEE AE E A rn eS nr nen ner en Ses E EE NE oe EO E 185 AKTA avant Operating Instructions 29 1015 56 AA 3 1 Introduction 1 Introduction About this chapter This chapter contains important user information descriptions of safety notices regula tory information intended use of the AKTA avant instrument and lists of associated documentation Section See page 1 1 About this manual 5 1 2 Important user information 6 1 3 Regulatory information 8 1 4 Associated Documentation 12 4 KTA avant Operating Instructions 29 1015 56 AA 1 Introduction 1 1 About this manual 1 1 About this manual Purpose of this manual The Operating Instructions provide you with the instructions needed to install operate and maintain the product in a safe way Typographical conventions Software items are identified in the text by bold italic text A colon separate
42. 90 to 700 nm 190 to 700 nm KTA avant Operating Instructions 29 1015 56 AA 3 System description 3 1 AKTA avant instrument overview Illustration of the main parts of the instrument The following illustration shows the location of the main parts of the instrument 4 P Part Function Function Fraction collector Buffer tray Instrument display Wet side Foldable door Pump cover Holder rails Swivel foot lock unlock knob Swivel foot Swing out toolbox Power switch Illustration of the wet side modules of the instrument The descriptions of the KTA avant instrument and the work flow in this manual are based on an KTA avant 25 instrument that consists of the modules and parts shown in the following illustration KTA avant 150 is delivered in a corresponding setup The following illustration shows the modules of the wet side of the instrument AKTA avant Operating Instructions 29 1015 56 AA 35 3 System description 3 1 AKTA avant instrument overview 13 12 Function Injection Valve 11 10 Function Flow Restrictor pH Valve Column Valve UV Monitor Conductivity Monitor Inlet Valve B Quaternary Valve System Pump B Pressure monitor of system pumps System pump flow restrictor Inlet Valve A System Pump A Mixer Sample pump flow restrictor Pressure monitor of sample pump Sample Pump Pump rinsing solution tube
43. AKTA avant Operating Instructions Original instructions Table of Contents Table of Contents 1 MoCU fe 9 ameeeeee ae eee eetercn near E 4 TM POM TAN Ole CAMO cassette tates cgece ates cca tecetep cdc E AAAA 5 1 2 mporn userin onma OW nicos an n E A N 6 la REJULO MONNA sit ce pease coer artes o aii aeea KEE eee eee 8 1 4 Associated DOCUMENtOtION cass se ecsesseccsncssncsectacotenusensscosncosvdanstblonsuncsunstevinnssbbjonesententiansseendenostiens 12 Salei NEU CHOS sesana 14 al Salei Fi MONS sesiyasida Esaa E a RENNRS ASNES 15 ea 60 2 eE E A E E E A A me TP 24 2 3 Emergency Procedures siriani re tomes E sae E E EE E E 21 PA REVENO MONA ON ess a 30 2 5 Declaration of Hazardous Substances DOHS ssssssssssssesesieireeserresssrseresessssssssssssssersererrrrrerrn 31 SVS em OESP OI assa 33 3 1 KTA AVANT instrument OVEFVICW s ss ssssssssssssssisssrsssorsssorsstrrosorssserssoorssotsssetrssorantrssonrsstnrssnrsn sens 34 e GS nN cs e 1 aeea nor EAE A rere ner me eerie eer 43 3 2 1 UNICORN software overview sesscsicsssnissssoterssnsvunsooneivaxocandiernnensivatenstossnsyvondoesseoaveideeusevnanssbensnennhs 44 3 2 2 The System Control module sssissrisriresesssssrrriricssisesisesiririsesriissssssesisririrererrsrennsses 46 o ON ar AAA 48 4l Sepe d TiO essien o Ee E E EEEE nanan 49 4 1 1 Dever vand SO Oe seere eee es 50 4 1 2 ROOM regulere WIS wicteisctistsrstrnaacrnaiidienddsnah aero EEEE 52 4 1 3 Si
44. G Protective ground The product must always be connected to a grounded power outlet Power cord Only use power cords with approved plugs deliv ered or approved by GE Access to power switch and power cord with plug Do not block access to the power switch and power cord The power switch must always be easy to access The power cord with plug must always be easy to disconnect Explosion hazard To avoid building up an explosive atmo sphere when using flammable liquids make sure that the room ventilation meets the local requirements Transportation route Doors corridors and elevators must have a minimum width of 75 cm to allow for trans porting the instrument Allow additional space for moving around corners 52 KTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 1 Site preparation 4 1 2 Room requirements Space requirements The following illustration shows the space recommended for the AKTA avant system Allow space on the laboratory bench for e handling of samples and buffers 2 x 30 cm e computer and monitor 80 cm e access for service see the following topic AKTA avant Operating Instructions 29 1015 56 AA 53 4 Installation 4 1 Site preparation 4 1 2 Room requirements Service access To access the rear panel the instrument can be rotated on a swivel foot There must be at least 20 cm additional space on the bench to allow for free rotation WARNING Rotating the
45. Part no 96 deep well plate GE 7701 5200 BD Biosciences 353966 Greiner Bio One 780270 Porvair Sciences 219009 Seahorse Bioscience S30009 Eppendorf 951033405 0030 501 306 48 deep well plate GE 7701 5500 Seahorse Bioscience S30004 24 deep well plate GE 7701 5102 Seahorse Bioscience S30024 AKTA avant Operating Instructions 29 1015 56 AA 63 4 Installation 4 2 Hardware installation 42 Hardware installation About this chapter This section describes the installation procedure of an AKTA avant system Note For information on how to how to unpack the AKTA avant instrument and how to lift the instrument onto a laboratory bench see AKTA avant Unpacking In Structions WARNING e Protective ground The product must always be connected to a grounded power outlet e Power cord Only use power cords with approved plugs deliv ered or approved by GE e Access to power switch and power cord with plug Do not block access to the power switch and power cord The power switch must always be easy to access The power cord with plug must always be easy to disconnect In this section This section contains the following subsections Section See page 4 2 1 Install the computer equipment 65 4 2 2 Connect the system units 66 4 2 3 Prepare waste tubing 70 4 2 4 Install the Barcode Scanner 2 D and the pH electrode 73 4 2 5 Prepare the pump rinsing system 74 4 2 6 Star
46. Prepare the system for a run 5 6 Calibrate the pH monitor 5 6 Calibrate the pH monitor Introduction If pH will be measured during the chromatographic run the pH monitor should be cali brated before the run is started Use two pH calibration buffers with a difference of at least one pH unit Preferably use a pH standard buffer pH 4 or 7 as the first calibration point and a pH standard buffer close to the lowest or highest pH you need to measure as your second point Allow the buffers to reach the operating temperature before use Note Do not run a system flow during pH calibration Calibrate the pH monitor CAUTION pH electrode Handle the pH electrode with care The glass tip may break and cause injury Follow the instructions to perform the calibration Step Action 1 Open the System Control module On the System menu click Calibration Result The Calibration dialog box opens Calibration System3 Monitor to calbvate iT S Curent value Calibration procedure Prepare for calibration Calibration for pH electrode buffer 1 m pH calibrates the pH electrode with two point PH for buffer 1 y calibration i e using two pH calibration solutions It can also be used to fill the pH cell with storage solution since the pH valve is in calibration position PH for buffer 2 For calibration preferably use pH standard buffers PH 4 or 7 as the first calibration point and a pH standard
47. V9 pH or V9H pH Valve which enables the pH electrode to be includ ed in the flow path or bypassed during a run The OH electrode may be calibrated when installed in the pH Valve It also enables the flow restrictor to be included in the flow path default position or bypassed during a run Outlet Valve V9 O or V9H O Valve which directs the flow to the fraction collec tor any of the ten outlet ports or waste UV Monitor U9 M Monitor which measures the UV Vis absorbance at up to three wavelengths simultaneously in the range 190 to 700 nm Conductivity Monitor C9 Monitor which continuously measures the conduc tivity of buffers and sample solutions Built in fraction collector Built in fraction collector A cooling function pro tects the fractions from heat degradation Core modules Core modules need to be installed for the system to run They are mandatory in the software All standard modules except the built in fraction collector are considered core modules 40 AKTA avant Operating Instructions 29 1015 56 AA Description of optional modules 3 System description 3 1 AKTA avant instrument overview The following modules may be added to the flow path Second Inlet Valve A and Inlet Valve B V9 A2 and V9 B2 or V9H A2 and V9H B2 Description Second inlet valve for System Pump A or System Pump B to extend the number of inlets up to 14 Inlet Valve X1 and Inlet Valve X2 V9 IX or V9H I
48. X Inlet valve with eight inlet ports No integrated air sensor Second Sample Inlet Valve V9 S2 or V9H S2 Second inlet valve for the Sample Pump to extend the number of sample inlets up to 14 Versatile Valve V9 V or V9H V A 4 port 4 position valve which can be used to cus tomize the flow path Loop Valve V9 L or V9H L Valve which enables automatic sample application from up to five sample loops or to collect intermediate frac tions in automated two step purification Second Column Valve V9 C2 or V9H C2 Valve which connects five additional columns to the in strument The valve allows the user to choose flow di rection through the column or to bypass the column Second Outlet Valve V9 O2 or V9H O2 Valve which adds 12 outlet ports to the system giving a total of 21 outlets Third Outlet Valve V9 O3 or V9H O3 Valve which adds 12 outlet ports to the system giving a total of 32 outlets External Air Sensor L9 1 5 or L9 1 2 Sensor which prevents air from being introduced into the flow path O box E9 Module which receives analog or digital signals from or transfers analog or digital signals to external equip ment that has been incorporated in the system Second UV Monitor U9 L Monitor which measures the UV absorbance at a fixed wavelength of 280 nm Second Conductivity Mon itor C9 Monitor which measures the conductivity of buffers and s
49. amaged After replacement perform a run to break in the new piston seal NOTICE Advanced maintenance Read the instruction carefully before disassembly of the pump head 160 KTA avant Operating Instructions 29 1015 56 AA 7 Maintenance 7 3 Replace pump piston seal O ring and rinse membrane of Pump P9 S Required material The following material is required e Adjustable wrench e Star screwdriver T20 e Cross headed screwdriver e Hex wrench e Ultrasonic bath e Ethanol 20 e Reference capillary Ref 1 e P9 S Seal kit 65 ml Illustration The illustration below shows the parts of the pump heads of Pump P9 S Description Description 7 9 Tad 1 2 3 4 Piston Piston seal Return spring Outlet check valve Drain plate O ring Rinse membrane Purge valve Rinse chamber Inlet check valve AKTA avant Operating Instructions 29 1015 56 AA 161 7 Maintenance 7 3 Replace pump piston seal O ring and rinse membrane of Pump P9 S Disassemble the pump head Step Action 1 2 162 Make sure that no run is in progress on the instrument Disconnect the tubing from the pump head and disconnect the pump inlet tubing Unscrew the two white plastic screws located below each pump head by hand Pull the plastic connectors to the sides to release the inlet manifold il m Disconnect the tubing of the pump piston rinsing system Unscrew one o
50. ample solutions Second Fraction Collector F9 R AKTA avant Operating Instructions 29 1015 56 AA Round fraction collector that can collect up to 175 fractions 41 3 System description 3 1 AKTA avant instrument overview Illustration of the instrument display The following illustration shows the instrument display with the system state Ready showing AKTAavant Ready Instrument display indicators and buttons The instrument display is a touchscreen that shows the current system status The in strument display includes the following indicators and buttons Indicator Button Description Indicates if the Instrument display buttons are unlocked or locked The buttons can be locked from UNICORN Sys tem Control Pauses the run and stops all pumps Resumes instrument operation from the following states panie e Wash e Pause e Hold 42 AKTA avant Operating Instructions 29 1015 56 AA 3 System description 3 2 UNICORN software 3 2 UNICORN software Introduction This section gives an overview of the UNICORN software It also describes the System Control module To learn more about System Control and the other three modules Administration Method Editor and Evaluation see the UNICORN documentation package In this section This chapter contains the following sections Section See page 3 2 1 UNICORN software overview 44 3 2 2 The System Control module 46
51. ave read and understood the Safety instructions chapter in this manual e Auser account has been created according to the UNICORN Administration and Technical Manual 6 AKTA avant Operating Instructions 29 1015 56 AA Safety notices 1 Introduction 1 2 Important user information This user documentation contains safety notices WARNING CAUTION and NOTICE concerning the safe use of the product See definitions below WARNING WARNING indicates a hazardous situation which if not avoided could result in death or serious injury It is important not to proceed until all stated conditions are met and clearly understood CAUTION CAUTION indicates a hazardous situation which if not avoided could result in minor or moderate injury It is important not to pro ceed until all stated conditions are met and clearly understood E NOTICE NOTICE indicates instructions that must be followed to avoid damage to the product or other equipment Notes and tips Note A note is used to indicate information that is important for trouble free and optimal use of the product Tip A tip contains useful information that can improve or optimize your procedures KTA avant Operating Instructions 29 1015 56 AA 7 1 Introduction 1 3 Regulatory information 1 3 Regulatory information Introduction This section lists the directives and standards that are fulfilled by the AKTA avant instru ment Man
52. buffers and samples to prepare which sample application technique to use that the pH electrode is connected if applicable which cassettes with corresponding deep well plates and or tubes to use in the fraction collector if applicable if it is a reversed phase chromatography RPC run KTA avant Operating Instructions 29 1015 56 AA 5 Prepare the system for a run 5 1 Before you prepare the system WARNING When using flammable liquids with the AKTA avant instrument follow these precautions to avoid any risk of fire or explosion Fraction collector Do not fractionate flammable liquids in the built in fraction collector When running RPC methods collect fractions through the outlet valve or the optional external Fraction Collector F9 R RPCruns with 100 acetonitrile and system pressure above 5 MPa 50 bar in AKTA avant 25 Always replace the green PEEK tubing between the used system pump and the pump pressure monitor with orange PEEK tubing i d 0 5 mm before running RPC with 100 acetonitrile Set the system pressure alarm to 10 MPa 100 bar RPC runs with 100 acetonitrile in AKTA avant 150 Always replace the beige PEEK tubing between the used system pump and the pump pressure monitor before running RPC with 100 acetonitrile Replace it with green PEEK tubing i d 0 75 mm AKTA avant Operating Instructions 29 1015 56 AA 103 5 Prepare the system for a run 5 2 Prepare the flowpath 5 2 Prepare the
53. couple of days or longer also perform the following e Fill the column with storage solution e g 20 ethanol using the Column CIP method pH electrode storage If pH monitoring will not be used for a week or longer perform one of the following ac tions e inject new storage solution into the pH flow cell e Replace the pH electrode with the dummy electrode that is installed in the pH valve on delivery In the following situations in order to increase the lifetime of the pH electrode use the By pass position and store the electrode in storage solution inside the pH flow cell e pH monitoring is not needed during the run e Organic solutions are used e Extremely acidic or extremely basic solutions are used For further information on how to prepare the pH electrode for storage refer to AKTA avant User Manual AKTA avant Operating Instructions 29 1015 56 AA 143 6 Runa method 6 5 After run procedures Log off or exit UNICORN Follow the instructions to log off or exit UNICORN This can be performed from any of the UNICORN modules If you want to log off UNICORN on the File menu click Log off Log off Eric Exit UNICORN Result All open UNICORN modules close and the Log On dialog box opens exit UNICORN on the File menu click Exit UNICORN Log off Eric Exit UNTCORN Result All open UNICORN modules close Note If an edited method or result is open and not saved when yo
54. ctions 29 1015 56 AA 127 6 Runa method 6 1 Before you start Warnings concerning use of hazardous substances WARNING e 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 mainte nance e Hazardous biological agents during run When using haz ardous 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 Hold pause or stop the run 128 At the end of a method the run stops automatically All pumps stop and an acoustic end signal sounds and End is displayed in the Run Log To interrupt a method during a run you may click the Hold Pause or End buttons in System Control A held or paused method run can be resumed by clicking the Continue button See the instructions in the following table If you want to temporarily hold the method with current click the Hold button flow rate and valve positions sustained 0 ee temporarily pause the method and stop click the Pause button all pumps AKTA avant Operating Instructions 29 1015 56 AA 6 Runa method 6 1 Before you start If you want to resume for example a held or paused click the Continue button method run Note An ended method cannot be continued
55. d make sure that the supply voltage at the wall outlet corresponds to the marking on the instrument KTA avant Operating Instructions 29 1015 56 AA 25 2 Safety Instructions 2 2 Labels Meaning This symbol indicates that the product might contain hazardous materials in excess of the limits established by the Chinese standard SJ T11363 2006 Requirements for Concentration Limits for Certain Hazardous Sub stances in Electronic Information Products The system complies with applicable European direc tives The system complies with applicable requirements for Australia and New Zealand 26 This symbol indicates that AKTA avant has been certi fied by a Nationally Recognized Testing Laboratory NRTL NRTL means an organization which is recog nized by the US Occupational Safety and Health Ad ministration OSHA as meeting the legal requirements of Title 29 of the Code of Federal Regulations 29 CFR Part 1910 7 AKTA avant Operating Instructions 29 1015 56 AA 2 Safety Instructions 2 3 Emergency procedures 2 5 Emergency procedures Introduction This section describes how to perform an emergency shutdown of the AKTA avant instru ment including connected equipment This section also describes the results in the event of power failure or network interruption Emergency shutdown In an emergency situation stop the run by either pausing the run or switching off the instrument as described in the f
56. d create reports e The default Evaluation module includes a user inter face 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 Classic it is possible to access descriptions of the active window by pressing the F1 key This can be especially helpful when editing methods KTA avant Operating Instructions 29 1015 56 AA 45 3 System description 3 2 UNICORN software 3 2 2 The System Control module 3 2 2 The System Control module Introduction The System Control module is used to start view and control a manual or method run System Control panes As seen in the following illustration three panes are shown in the System Control module by default The Run Data pane 1 presents current data in numerical values The Chromatogram pane 2 illustrates data as curves during the entire run The current flow path is illustrated in the Process Picture 3 which allows manual inter actions with the system and provides feedback on run parameters B System Control Lo File Edit View Manual System Tools Help ES le O u Buy E E _ 1012 047 X Run Data x A System state Acc volume System flow Sample flow PreC pressure DeltaC pressure UV 1_280 UV 2nd pH
57. d is har ples for design Risk assessmentand monized with EU direc risk reduction tive 2006 42 EC EN IEC 61010 1 Safety requirements for electrical EN standard is harmo UL 61010 1 equipment for measurement control nized with EU directive CAN CSA C22 2 and laboratory use 2006 95 EC No 61010 1 EN IEC 61326 1 Electrical equipment for measure EN standard is harmo Emission accord Ment control and laboratory use nized with EU directive ing to CISPR 11 EMC requirements 2004 108 EC Group 1 class A ETSI EN 301 489 3 Electromagnetic compatibility and EN standard is harmo Radio spectrum Matters ERM Elec nized with EU directives troMagnetic Compatibility EMC 1999 5 EC standard for radio equipment and Services AKTA avant Operating Instructions 29 1015 56 AA 9 1 Introduction 1 3 Regulatory information Standard Description ETSI EN 300 330 2 Electromagnetic compatibility and EN standard is harmo Radio spectrum Matters ERM Short nized with EU directive Range Devices SRD Radio equip 1999 5 EC ment in the frequency range 9 kHz to 25 MHz and inductive loop sys tems in the frequency range 9 kHz to 30 MHZ FCC compliance This device complies with part 15 of the FCC Rules Operation is subject to the following two conditions 1 This device may not cause harmful interference and 2 this device mustacceptany interference received including interference that may cause undesi
58. dations stated in this section General computer specifications The table below describes the recommended computer specifications for a UNICORN system operating with AKTA instruments Installation is supported for Windows 7 Profes sional 32 bit or 64 bit with Service Pack 1 Workstation E License Server installation UNICORN Client Database Server Min free disk space Min available RAM 3 GB 3 GB 3 GB 2 GB Disc format NTFS NTFS NTFS NTFS OS Windows 7 Professional SP1 32 64 bit Windows 7 Professional SP1 32 64 bit Windows Server 2008 R2 64 bit Windows 7 Professional SP1 32 64 bit Windows 7 Professional SP1 32 64 bit Windows Server 2008 R2 64 bit OS language Architecture English U S Code 1033 Intel Dual Core or faster English U S Code 1033 Intel Dual Core or faster English U S Code 1033 Intel Dual Core or faster English U S Code 1033 Intel Dual Core or faster Note e UNICORN is tested using an English operating system version Using other language versions of the operating system may cause errors e A screen resolution of 1280x1024 or higher is recommended Parts of the UNICORN user interface may not be displayed properly using a lower res olution e Changing the default font and font size in Windows may cause problems in the UNICORN user interface AKTA avant Operating Instructions 29 1015 5
59. detailed information about the maintenance procedures Maintenance is divided into e Daily maintenance e Weekly maintenance e Monthly maintenance e Bi annual maintenance e Maintenance when required WARNING Electrical shock hazard All repairs should be done by service personnel authorized by GE Do not open any covers or replace parts unless specifically stated in the user documentation Periodic maintenance program The following periodic maintenance should be performed by the user of the AKTA avant instrument Interval Maintenance action Daily Calibrate the pH monitor Weekly Change pump rinsing solution Weekly Replace inline filter in the mixer Weekly Clean the fraction collector Monthly Check the flow restrictor Bi annual Clean the UV flow cell Bi annual Replace the pH electrode 146 KTA avant Operating Instructions 29 1015 56 AA 7 Maintenance 7 1 Maintenance program Maintenance when required The following maintenance should be performed by the user of the AKTA avant instrument when required Refer to the AKTA avant User Manual for detailed instructions Maintenance action Clean the instrument externally Run System CIP System cleaning in place Run Column CIP Column cleaning in place Clean the fraction collector Replace tubing and connectors Storage of pH electrode Clean the pH electrode Clean the conductivity flow cell
60. e Sample loop or Superloop Column Valve Column UV Monitor Conductivity Monitor pH valve with pH monitor Flow Restrictor Outlet Valve Fraction collector Prepare the inlet tubing Connect inlet tubing to the inlet ports that are to be used and immerse all inlet tubing that is to be used during the method run in the correct buffers AKTA avant Operating Instructions 29 1015 56 AA 5 Prepare the system for a run 5 2 Prepare the flowpath Waste ports The following table shows the waste ports of Injection Valve pH Valve and Outlet Valve Valves and ports Illustrations Injection Valve label V9 Inj and V9H Inj Waste ports e W1 W2 pH Valve label V9 pH and V9H pH Waste port e W3 Outlet Valve label V9 O and V9H O Waste port e W KTA avant Operating Instructions 29 1015 56 AA 107 5 Prepare the system for a run 5 2 Prepare the flowpath Prepare the waste tubing Make sure that the waste tubing is prepared according to the instructions in Section 4 2 3 Prepare waste tubing on page 70 Prepare the outlet tubing Connect outlet tubing to the outlet ports of the outlet valve that are to be used during the run If a fraction collector is to be used make sure that tubing is connected between the outlet valve Frac port and the fraction collector and prepare the fraction collector Otherwise immerse th
61. e Documentation Main contents UNICORN Help Descriptions of UNICORN dialog boxes available from the Help menu 12 KTA avant Operating Instructions 29 1015 56 AA 1 Introduction 1 4 Associated Documentation Documentation Main contents Getting started with Video clips showing common workflows in the Evaluation Evaluation module Note Overview of features of the Evaluation module Available in UNICORN 7 0 and later UNICORN Method Manual Overview and detailed descriptions of the method creation features in UNICORN Workflow descriptions for common operations UNICORN aie Overview and detailed description of network and Technical Manual setup and complete software installation Administration of UNICORN and the UNICORN database a Evaluation Overview and detailed descriptions of the Manual Evaluation Classic module in UNICORN Description of the evaluation algorithms used in UNICORN ee System Control Overview and detailed description of the system Manual control features in UNICORN Includes general operation system settings and instructions on how to perform a run 1 Current UNICORN version is added to the title of the manual AKTA avant Operating Instructions 29 1015 56 AA 13 2 Safety Instructions 2 Safety Instructions About this chapter This chapter describes safety precautions and emergency shutdown procedures for the product The labels on the system and informatio
62. e O ring on the purge valve with a new O ring 3x1 mm and screw the purge valve back into the pump head Note Always use Lubricant 56686700 when exchanging the O ring 3 x 1 mm AKTA avant Operating Instructions 29 1015 56 AA 7 Maintenance 7 3 Replace pump piston seal O ring and rinse membrane of Pump P9 S Step Action 2 Wet a new seal with 20 ethanol Place the new seal in the hole in the front section of the pump head and press it into position 3 With the pump head facing downwards on the bench place the rinse chamber onto the front section of the pump head with the rinse ports in line with the check valves The conical depression in the rinse chamber shall be facing upwards Wet a new membrane with 20 ethanol and place the membrane into the rinse chamber with the conical face upwards Assemble the pump head Make sure to assemble the pump head correctly Refer to ustrations on page 150 Step Action 1 Place the drain plate on top of the assembly Screw the two screws through the drain plate and the rinse chamber using a cross headed screwdriver KTA avant Operating Instructions 29 1015 56 AA 165 7 Maintenance 7 3 Replace pump piston seal O ring and rinse membrane of Pump P9 S Step Action e Wipe clean the piston and remove all finger prints Wet the piston with 20 ethanol and insert the piston into the return spring With the pump head facing downwards on the bench insert the piston
63. e enuirOnmEN scart tastcartse aspciovcnu utara EAE al es te ee 56 4 1 4 Power PERU CLIC E maseerssnrnkhinro ninn A E Ee 57 4 1 5 Computer requirements eccsesssessssseessesssessesssecsuessscssscsusessssucssecsuscsscssscsseasecsuessscasecauscseeseesecesecses 59 4 1 6 kegu a mO aea AE NINE ANAE AEE E EO 61 AC n NO an EEE 64 4 2 1 Install the computer egqUIPMEnt aeccsz secs secs ssczscceszcecac ucts coos ees etecassceectg sa cosssetasetase tape teebseeebees esate 65 4 2 2 Connect the systemi UNIS assisas EEE E ESA A EN REEE EE aE 66 4 2 3 Fel e118 1 Coat U0 4 TUONO sea A A 70 4 2 4 Install the Barcode Scanner 2 D and the PH electrode ou eeeeseecsesssessssessssessseesseeesseessnes 73 4 2 5 Prepare the pump rinsing SYSTE misasi 74 4 2 6 Start the instrument and the COMPUTE ssssssssessssssssirirsssssssssssssssrsrrirrssssssssssrrrsrrrrnnnrs 77 4 3 Software installation ass czsontnstescecutssossectvzvonngsuvennoststvedscestespessSbtidesvvssiesSscannssnsseesneste uadeemieounis 78 Ad Start UNICORN and connect to System ssssssssssssssssssssssssssssssssssssseserssresrssrirsrrrirerrierrrerrnrrnnrnness 79 4 5 Prime inlets and purge pump headS cei cess occa cacesbesiacoastdivensuncd dveaceuessninete Resteddpbvahavtonssers 82 4 5 1 Prime buffer inlets and purge system puUmpsS scvssc cccssseciucsstesscsssdsuschssvenuvsctsauovicvhiaciewadeass 83 4 5 2 Prime sample inlets and purge Sample PUMP ueeseesssessssessssesssessssessssecssesssesssessss
64. e outlet tubing in suitable tubes or flasks Plug unused valve ports It is recommended to plug all unused valve ports with stop plugs before starting a run See AKTA avant User Manual for information about connectors 108 AKTA avant Operating Instructions 29 1015 56 AA 5 Prepare the system for a run 5 3 Prime buffer inlets and purge system pumps 5 3 Prime buffer inlets and purge system pumps Introduction Before you start the system pumps it is important to do the following e Prime the inlets fill the buffer inlets with liquid e Purge the system pumps remove air from the pump heads For instructions on how to prime the inlets and purge the system pumps see Section 4 5 Prime inlets and purge pump heads on page 82 AKTA avant Operating Instructions 29 1015 56 AA 109 5 Prepare the system for a run 5 4 Connect a column 54 Connect a column Introduction This section describes how to connect a column to the instrument using a column holder and without introducing air into the flow path Several types of column holders are available for the KTA avant instrument WARNING Before connecting to a column read the instructions for use of the column To avoid exposing the column to excessive pressure make sure that the pressure limit is set to the specified maximum pressure for the column Methods automatically include a pressure alarm based on the specifications of the chosen column type Howev
65. e priming of Q inlet tubing Purge Quaternary Valve and the system pumps if pressure signal indicates air bubbles Validate purge of Quarternary Valve and system pumps End the run Prime the Q inlets Follow the instructions to prime the Q inlets Step Action 1 Make sure that the pieces of inlet tubing marked A1 B1 and Q1 Q4 are im mersed in the correct buffers The A1 and B1 positions are used for pump synchronization and these lines should already be primed AKTA avant Operating Instructions 29 1015 56 AA 95 4 Installation 4 5 Prime inlets and purge pump heads 4 5 3 Prime Q inlets Step Action 2 In the Manual instructions dialog box e Select Pumps and pressures Quaternary start concentrations e Set Start concentration Q1 to 100 Make sure that the other start concentrations are set to 0 Manual instructions System3 Selected column type L Select Instructions Parameters for Quaternary start concentrations Pumps and pressures e Start concentration 01 0 0 100 0 System flow Be Sample flow 100 0 5 Gradient Start concentration Q2 0 0 100 0 Pump wash as System wash 0 0 2 Quaternary start concentrations Start concentration 03 0 0 100 0 Quaternary gradient 2 BufferPro pH E 0 0 a _ __ Column packing flow Start concentration 04 0 0 100 0 Flow path Bx H Monitors a 0 0 Save result as Browse _ Auto
66. ecsssecsseess 90 4 5 5 Prime Q inlees seses EA AAEE EEE E AAE 95 46 PS TRO NIC teste suuosa A tease ada eens 100 Prepare the system fOr a run cscccsscessessssscscssscssscessseseccessessseesseessesesseesseess 101 5 1 Before YOU prepare the SYSTEM ou ceeecesssesssesssessseessesssecssecsssssssessuecsuecssecssecsseessessssessueessessseesseessees 102 oh A oy gt 0 gt 20910 11 1 810 a E ETA oer CerT ae 104 AKTA avant Operating Instructions 29 1015 56 AA Table of Contents 5 3 Prime buffer inlets ANd purge SYSTEM PUMPS ceccseesssesssessseessssesseessecsseessecssecssesseessseesseessees 109 54 COnnecta COUN seserian AEAEE EERS 110 5 5 Set pressure 6 S eae naan ne iee NAE AR En E OTE ee eR 115 x6 Caliprate the pH MONIO sieniin RAA 117 57 Prepare tne puit miracUon Colector seerste ari 119 5 8 Prepare for a run at cold temperature s sssssssssiisiiisieririirrirrrsrrsrrrsrrsssssssssssssssssssssssssssssrssses 125 C Fona mN n E wees ee eames 126 e EE 21 6 Vou T a E E O E aoe 127 62 PUTAS SOMPE sarisini cen see E EERE 130 OVI TS O T E EEE E 133 I CUMIN E TUN eenn ieo aA E E E 139 os Mammy 110 1 9 OC O e E AA 142 T PONNE aaia E as 145 7 1 OTN INC Progr aiT ssispreedenaniiiarth s retien asne EE EEEN ESAE 146 7 2 Replace pump piston seal O rings and rinse membrane of Pump P9 or Pump BT cxestaa E eset ciara Soars E OA vaste AT N A A A 149 7 3 Replace pump piston seal O ring and rinse membrane of Pump P
67. em Settings are described in KTA avant User Manual AKTA avant Operating Instructions 29 1015 56 AA 119 5 Prepare the system for a run 5 7 Prepare the built in fraction collector Prepare and insert the cassette tray Follow the instructions to prepare the fraction collector before a run Cassettes and cassette tray Step Action 1 If you are to use cassettes with the QuickRelease function first open the cassettes 2 Place the tubes and deep well plates in the cassettes Make sure that the deep well plates are rotated so that the well marked A1 is positioned above the A1 marking on the cassette 120 AKTA avant Operating Instructions 29 1015 56 AA 5 Prepare the system for a run 5 7 Prepare the built in fraction collector Step Action Close the cassettes that have the QuickRelease function Place the cassettes on the cassette tray Make sure that the cassette type code see the illustration faces the front of the tray marked with the GE monogram Open the fraction collector drawer by pressing the handle upwards and 5 pulling out the drawer KTA avant Operating Instructions 29 1015 56 AA 5 Prepare the system for a run 5 7 Prepare the built in fraction collector Step Action 6 Place the cassette tray on the tray support of the fraction collector drawer Make sure that the front of the tray marked with the GE monogram faces the front of the drawer and is hooked onto the two pins
68. end the run inal click the button Note When ending a method run in advance it is possible to save the partial result More information regarding UNICORN capabilities during the method run is available in the UNICORN System Control Manual AKTA avant Operating Instructions 29 1015 56 AA 139 6 Runa method 6 4 Monitor the run Process Picture The Process Picture displays the current flow path run parameters and real time data from monitors during a run It also allows manual interactions with the system Tubing colors indicate flow path states as shown in the following illustration and de scribed in the following table Modules without a fixed place in the system are shown in a panel below the process picture modules are called components in the process picture System flow 1 000 mi min 0 0 8 Sample flow 0 000 Prec MPa Co S c gt Manual load rec 0 23 MPa Cond 0 05 mS cm pH Off Temp 20 5 C de High alarm 0 50 uvi 593 845 mau EEANN mu A1 E A 2 E w1 e tr OY P M E SyP F 1 a UV mmm Cond mmm Restrictor mmm Frac mum EE Et pa A es Baas a Bi E r EET EIA f Deltac 0 01 MPa High alarm 0 30 f Bulfin S i P A Available components without a fixed place 43 839 mau UV 2nd 1 0 box Indication Open flow path with flow Closed flow path or an open path without flow Syringe port in loop open for manual injection 140 KTA avant Operating Instructions 29 1015 56 AA 6 Runa me
69. ep 26 10 Desalting Column Handling Scouting run 3 HiPrep 26 10 Desalting Result The run starts All necessary actions occur automatically according to the method including ending of the run 138 AKTA avant Operating Instructions 29 1015 56 AA 6 Runa method 6 4 Monitor the run 6 4 Monitor the run Introduction You may follow the on going method run in the System Control module The current system status is shown in the System state panel in the Run Data pane For example it may state Run Wash or Hold The same information is also shown on the Instrument display e Selected curves are shown in the Chromatogram pane e All registered actions during the run are displayed in the Run Log pane e The current flow path is shown in the Flow Scheme pane For an overview of the System Control interface see Section 3 2 2 The System Control module on page 46 Monitor the run To interrupt a method during a run you may click the Hold Pause or End buttons in System Control A held or paused method run can be resumed by clicking the Continue button See the following table If you want to temporarily hold the method with current ON flow rate and valve positions sustained click the It button temporarily pause the method and stop all pumps A E click the button resume for example a held or paused ra method run click the lt 4 button Note An ended method cannot be resumed permanently
70. er when running manual runs you have to set the pressure limits yourself Also to protect the column media special settings are needed See Sec tion 5 5 Set pressure alarms on page 115 for more information on pressure alarms Note Do not overtighten when connecting columns Overtightening might rupture the connectors or squeeze the tubing and thereby result in high back pressure 110 AKTA avant Operating Instructions 29 1015 56 AA 5 Prepare the system for a run 5 4 Connect a column Attach a column holder and connect a column Follow the instructions to connect a column to the instrument Always use a column holder The column is connected to two opposite parts of the column valve using appro priate tubing and connectors Step Action 1 Attach an appropriate column holder to the rail on the instrument AKTA avant Operating Instructions 29 1015 56 AA 111 5 Prepare the system for a run 5 4 Connect a column Step Action 3 Connect a suitable tubing to a column valve port for example port 1A if column position 1 was chosen in the method to be run 4 In the Process Picture e Click the Column valve icon e Click e g Position 1 and Down flow Result The column valve switches to position 1 112 AKTA avant Operating Instructions 29 1015 56 AA 5 Prepare the system for a run 5 4 Connect a column Step Action 5 In the Process Picture e Click the System pumps icon e Enter alow System flow
71. f the selected system pump Make sure that the syringe fits tightly into the purge connector 98 AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 5 Prime inlets and purge pump heads 4 5 3 Prime Q inlets Step Action 5 Open the purge valve by turning it counterclockwise about 3 quarters of a turn Draw 10 ml of liquid slowly into the syringe with a rate of about 1 ml per second 6 Close the purge valve by turning it clockwise Disconnect the syringe and discard its contents 7 Repeat steps 3 to 5 for the other three purge valves of the system pumps to get rid of air in all pump heads Keep the system flow running during this procedure 8 Check that there is no air left in the pump by following the instructions in Validate prime or purge of System Pump A or B or Sample Pump on page 89 End the run Click the End button in the System Control toolbar to end the run mans KTA avant Operating Instructions 29 1015 56 AA 99 4 Installation 4 6 Performance tests 46 Performance tests Introduction Before taking the AKTA avant instrument into use run performance tests to check the function of the equipment See AKTA avant User Manual for further instructions 100 AKTA avant Operating Instructions 29 1015 56 AA 5 Prepare the system for a run 5 Prepare the system for a run About this chapter This chapter describes the preparations necessary before starting a run In thi
72. f the two screws of the pump head using a hex wrench Un screw the second screw and at the same time push firmly on the front of the rinse chamber to compensate for the pressure of the piston return spring KTA avant Operating Instructions 29 1015 56 AA 7 Maintenance 7 3 Replace pump piston seal O ring and rinse membrane of Pump P9 S Step Action 6 Place the pump head face down on the bench Pull out the piston together with the return spring 7 Inspect the piston and return spring for sign of damage If damaged discard the piston and return spring and use a new piston and return spring when assembling the pump head 8 Unscrew the two screws securing the drain plate and the rinse chamber Lift off the drain plate and discard the membrane located between the drain plate and the rinse chamber TI AKTA avant Operating Instructions 29 1015 56 AA 163 7 Maintenance 7 3 Replace pump piston seal O ring and rinse membrane of Pump P9 S Step Action 9 10 Lift off the rinse chamber Gently pull off the piston seal Discard the used seal Clean the pump head rinse chamber and drain plate in an ultrasonic bath If there are particles on any surfaces the check valves should be removed and cleaned separately see Section 7 5 Clean pump head check valves on page 170 Replace O ring piston seal and rinse membrane Step 1 164 Action Unscrew the purge valve of the pump head Replace th
73. fer Inlet valve B e 81 B2 83 B4 B5 B6 87 Result The inlet valve switches to the selected port Connect a 25 to 30 ml syringe to the purge valve of one of the pump heads of System Pump B Make sure that the syringe fits tightly into the purge connector oa Te Wii Open the purge valve by turning it counter clockwise about three quarters of a turn Draw liquid slowly into the syringe until the liquid reaches the pump Close the purge valve by turning it clockwise Disconnect the syringe and discard its contents Repeat steps 3 to 6 for each piece of inlet tubing that is to be used during the run In the final inlet position draw liquid into the syringe through both purge valves AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 5 Prime inlets and purge pump heads 4 5 1 Prime buffer inlets and purge system pumps Step Action 8 Check that there is no air left in the pump by following the instructions in Validate prime or purge of System Pump A or B or Sample Pump on page 89 If air bubbles are indicated follow the instructions in Purge System Pump B on page 85 Purge System Pump B If the priming was done thoroughly and the final buffer was drawn all the way into the syringe and the validation of the priming showed that there was no air left in the pump it is not necessary to purge System Pump B However if the pressure signal indicated air bubbles left in t
74. fore continuing with the next instruction Maintenance interval Replace the pump pistons if they are damaged Required material The following materials are required e Adjustable wrench e Star screwdriver T20 e Piston kit 168 KTA avant Operating Instructions 29 1015 56 AA 7 Maintenance 7 4 Replace pump pistons Replace pump pistons of Pump P9 and P9H If a damaged piston has been in operation the piston seal will be destroyed and should also be replaced To replace the piston and the seal of a system pump see Section 7 2 Replace pump piston seal O rings and rinse membrane of Pump P9 or Pump P9H on page 149 Section 7 2 Replace pump piston seal O rings and rinse membrane of Pump P9 or Pump P9H on page 149 Replace pump pistons of Pump P9 S If a damaged piston has been in operation the piston seal will be destroyed and should also be replaced To replace the piston and the seal of Pump P9 S see Section 7 3 Replace pump piston seal O ring and rinse membrane of Pump P9 S on page 160 AKTA avant Operating Instructions 29 1015 56 AA 169 7 Maintenance 7 5 Clean pump head check valves 7 5 Clean pump head check valves Introduction Clean the check valves when required for example if particles like dust or salt crystals in the check valve cause irregular or low flow The cleaning procedure is the same for the system pumps and the sample pump Required material The following materials are
75. he KTA avant instrument in any other way than described in the AKTA avant and UNICORN manuals e Only properly trained personnel may perform operation and user maintenance of the product e Before connecting to a column read the instructions for use of the column To avoid exposing the column to excessive pressure make sure that the pressure limit is set to the speci fied maximum pressure for the column AKTA avant Operating Instructions 29 1015 56 AA 15 2 Safety Instructions 2 1 Safety Precautions WARNING e Do not use any accessories not supplied or recommended by GE e Do not use KTA avant if it is not working properly or if it has suffered any damage for example damage to the power cord or its plug damage caused by dropping the equipment damage caused by splashing liquid onto it NOTICE Avoid condensation If KTA avant is kept in a cold room cold cabinet or similar keep it switched on in order to avoid condensa tion Flammable liquids and explosive environment WARNING When using flammable liquids with the AKTA avant system follow these precautions to avoid any risk of fire or explosion e Fire Hazard Before starting the system make sure that there is no leakage e Explosion hazard To avoid building up an explosive atmo sphere when using flammable liquids make sure that the room ventilation meets the local requirements e Fraction collecto
76. he pump follow these in structions to purge both pump heads of System Pump B Step Action 1 Make sure that the piece of waste tubing connected to the injection valve port W1 is placed in a waste vessel 2 In the Process Picture e Click the Injection valve icon and then click System pump waste Result The injection valve switches to waste position This is necessary to achieve a low back pressure during the purge procedure System pump waste Loope Loop valve pret System pump By pass waste wi i om syp Injection valve Manual load j System Inject Direct inject Sample pump waste J KTA avant Operating Instructions 29 1015 56 AA 85 4 Installation 4 5 Prime inlets and purge pump heads 4 5 1 Prime buffer inlets and purge system pumps Step Action 3 In the Process Picture e Click the Inlet valve B icon e Click the position of one of the inlets that will be used at the beginning of the run Yi od B1 r a Inlet valve B x B2 B3 B4 B5 B 6 B7 Result The inlet valve switches to the selected port 4 In the Process Picture e Click the System pumps icon e Set Conc B to 100 B and click Set B System flow 4 1 000 mimin Set flow rate Conc B Pump wash A B Al v Start pump A wash B1 v Start pump B wash Qi v Start Q inlet wash System wash System out W Injection valve W 1 15
77. install the module in the instrument if applicable 184 AKTA avant Operating Instructions 29 1015 56 AA Index A AKTA avant preparation 119 AKTA avant system specifications 174 C CE conformity 8 CE marking 9 Cleaning column 143 Cold room precautions 125 Column cleaning 143 pressure alarm 115 storage 143 Connect system units 66 D Documentation 12 E Emergency procedures emergency shutdown 27 power failure 28 Explosive environment precautions 16 F FCC compliance 10 Flammable liquids precautions 16 Fraction collector requirements on deep well plates 62 requirements on tubes 62 G General precautions 15 Inlet tubing prime inlet tubing B 83 KTA avant Operating Instructions 29 1015 56 AA Index Installation software 78 Installing and moving precau tions 18 Instrument overview 34 modules 37 L Labels safety symbols 25 system label 25 Log on UNICORN 79 M Maintenance precautions 22 Maintenance program 146 Manufacturing information 8 N Node ID change Node ID 184 check Node ID 184 description 181 Notes and tips 7 O Outlet tubing prepare 108 P Personal protection 17 18 pH monitor storage of the pH elec trode 143 Prepare the system KTA avant 119 Start UNICORN 79 Pressure alarm set 115 Prime inlet tubing B 83 Prime sample inlets 90 Process picture 140 Pumps clean check valves 170 185 Index
78. instructions 22 KTA avant Operating Instructions 29 1015 56 AA 2 Safety Instructions 2 1 Safety Precautions WARNING e Electrical shock hazard All repairs should be done by service personnel authorized by GE Do not open any covers or replace parts unless specifically stated in the user documentation e Disconnect power Always disconnect power from the instru ment before replacing any component on the instrument un less stated otherwise in the user documentation e Corrosive chemicals during maintenance If the system or column is cleaned with a strong base or acid flush with water afterwards and wash with a weak neutral buffer solution in the last step or phase NOTICE To avoid damage to the KTA avant instrument or other equipment when performing maintenance on the KTA avant instrument follow these instructions e Cleaning Keep the exterior of the instrument dry and clean Wipe regularly with a soft damp tissue and if necessary a mild cleaning agent Let the instrument dry completely before use e Advanced maintenance Read the instruction carefully before disassembly of the pump head KTA avant Operating Instructions 29 1015 56 AA 23 2 Safety Instructions 2 2 Labels 2 2 Labels Introduction This section describes the safety labels that are attached to the AKTA avant instrument For information about marking of the computer equipment refer to the manufacturer s
79. instructions Labels on the AKTA avant instrument The following illustrations show the labels that are attached to the AKTA avant instrument N3732 L T Made in Sweden Conforms to ANSI UL Std 61010 1 Cert to CAN CSA 24 KTA avant Operating Instructions 29 1015 56 AA 2 Safety Instructions 2 2 Labels System label Note The specific data shown on this system label is only an example Actual data is specific for each individual system and may vary from system to system 28930842 AKTA avant 25 Code no 28924630 Voltage 100 240 V Serial no Frequency 50 60 Hz Mfg Year Max Power 800 VA Protection Class Gy Wet Side IP22 V LEDER ms TT GE Healthcare Bio Sciences AB Made in Sweden 751 84 Uppsala Sweden B Conforms to ANSI UL Std 61010 1 Cert to CAN CSA Std C22 2 No 61010 1 Safety symbols The following safety symbols are used in the labels Meaning Warning Fraction collector Do not fractionate flammable liq uids in the built in fraction collector When running RPC methods collect fractions through the outlet valve or the optional external Fraction Collector F9 R Warning Read the Operating Instruction before using the system Electrical shock hazard All repairs should be done by service personnel authorized by GE Do not open any covers or replace parts unless specifically stated in the user documentation Supply voltage Before connecting the power cor
80. into the pump head by pushing it gently but firmly vertically downwards into the seal Place the complete pump head over the locating pins on the front panel of the sample pump module Turn the pump head so that the text UP on the drain plate is facing upwards Push firmly on the front of the pump head and at the same time screw one of the screws to fasten the pump head onto the front of the module using a hex wrench Screw the second screw of the pump head Make sure to tighten both screws fully Reconnect the tubing of the pump piston rinsing system Reconnect the inlet manifold Reconnect the tubing to the pump head and reconnect the pump inlet tubing Break in the new pump piston seal see instruction below Break in the new pump piston Follow the instruction to break in the new pump piston seal of Pump P9 S Action seal Step 1 2 3 4 166 Fill a buffer vessel with 20 ethanol in water Immerse a piece of sample inlet tubing for example S1 in the buffer vessel Place the buffer vessel on the buffer tray Prime the inlets and purge the pump see Section 4 5 2 Prime sample inlets and purge Sample Pump on page 90 Connect the reference capillary Ref 1 or an equivalent capillary that gives a backpressure of 2 to 3 MPa 20 to 30 bar to one of the column positions of the column valve e g ports 1A and 18 Immerse the waste tubing in the buffer vessel to recirculate the liquid AKTA avant Operating Instruct
81. ions 29 1015 56 AA 7 Maintenance 7 3 Replace pump piston seal O ring and rinse membrane of Pump P9 S Step Action 5 In the Manual instructions dialog Select Flow path Column position and select the Position of the capillary connected to the column valve Click Insert Select Flow path Sample inlet and select a Position Click Insert Select Flow path Injection valve and click Direct inject on the Position menu Click Insert Select Pumps Sample flow and set the Flow rate to 25 0 ml min Click Insert Click Execute Result A sample flow of 25 0 ml min starts 6 Run the flow for 2 hours 7 Discard the used buffer KTA avant Operating Instructions 29 1015 56 AA 167 7 Maintenance 7 4 Replace pump pistons 7 4 Replace pump pistons Introduction Follow the instructions to replace the pump pistons of the pumps P9 P9H and P9 S Note Always replace the O rings piston seals and rinse membranes of both pump heads of a pump at the same time NOTICE e Donot disassemble the pump head unless there is good reason to believe that there is an internal leakage A sign of leakage is increasing pump rinsing solution volume Always make sure that sufficient spare components are available before attempt ing to replace a spare part e Replacing spare parts Read the instructions carefully For example some individual parts of the pump head can be as sembled incorrectly Check the orientation of each part be
82. itanium PEEK and highly resistant fluoropoly mers and fluoroelastomers Titanium is used as far as possible to minimize contribution of potentially deactivating metal ions such as iron nickel and chromium There is no standard stainless steel in the flow path Plastics and rubber materials are selected to avoid leakage of monomers plasticizers or other additives Cleaning chemicals Strong cleaning works well with 2 M sodium hydroxide 70 acetic acid or the alcohols methanol ethanol and isopropyl alcohol Complete system cleaning using 1 M hydrochlo ric acid should be avoided in order to not damage the pressure sensors If you are cleaning separation media using 1 M hydrochloric acid use loop injections of the acid and make sure that the column is not mounted on the Column Valve V9 C The Column Valve V9 C contains a pressure sensor which can be damaged by 1 M hydrochloric acid Long time use of 0 2 M HCI connected to the Quaternary Valve Q9 as part of a BufferPro recipe is acceptable The solution becomes diluted further down in the system If sodium hypochlorite is used as sanitizing agent instead of 2 M sodium hydroxide use a concentration up to 10 Organic solvents 176 Reversed phase chromatography of proteins works well with 100 acetonitrile and ad ditives trifluoroacetic acid TFA up to 0 2 or formic acid up to 5 Strong organic solvents like ethyl acetate 100 acetone or chlorinated organic solvents should be av
83. letely emptied AKTA avant Operating Instructions 29 1015 56 AA 6 Runa method 6 3 Start a method run 6 3 Starta method run Introduction This section describes how to start a run using a previously created method If Column Logbook was enabled during installation of the software registration and selection of individual columns is possible at method start For further information on method creation please refer to UNICORN Method Manual Choose and start a method The following instructions describe how to open a method and start a run Step Action 1 Open the System Control module and click the Open Method Navigator button E Result Tne Method Navigator pane opens Method Navigator ax amp Methods Method fe Folder name System Last modified Created by DoeMethod 2014 10 06 14 3 Default Bevis 2012 11 27 14 5 Default H AutoTests 2013 05 31 2013 05 31 12 0 Default E Evis 2012 11 27 14 5 Default 2 Select the method to run and click the Run button Result The Start Protocol dialog box opens 3 Step through the displayed pages in the Start Protocol add requested input and make appropriate changes if necessary Click Next KTA avant Operating Instructions 29 1015 56 AA 133 6 Runa method 6 3 Start a method run Step Action 4 Click Start on the last page of the Start Protocol Result e If column logging was chosen at installation of UNICORN and
84. liance of connected equipment Any equipment connected to AKTA avant should meet the safety requirements of EN IEC 61010 1 or relevant harmonized standards Within EU connected equipment must be CE marked AKTA avant Operating Instructions 29 1015 56 AA 11 1 Introduction 1 4 Associated Documentation 14 Associated Documentation Introduction This section describes the user documentation that is delivered with the AKTA avant in strument AKTA avant user documentation The user documentation listed in the following table is delivered with the AKTA avant instrument Document Main contents KTA avant Unpacking Instruction Instructions for unpacking the instrument and how to lift the instrument onto a bench KTA avant Operating Instructions Instructions needed to install operate and maintain the system in a safe way KTA avant User Manual Instructions for handling the system Descrip tions of components Information about how to run and maintain the system KTA avant 25 Product System specification and declaration of material Documentation conformity OR KTA avant 150 Product Documentation t 1 The instrument is delivered with the relevant document 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 modul
85. ll plates WARNING Moving parts in fraction collector Do not open the fraction collec tor drawer when the fraction collector is active If you need to ac cess the fraction collector press Pause and make sure that the movement has stopped before opening the drawer AKTA avant Operating Instructions 29 1015 56 AA 123 5 Prepare the system for a run 5 7 Prepare the built in fraction collector View fraction collector content To view the content of the Fraction collector open the System control module On the View menu click Fraction Collector Content Fraction Collector Content System3 OPP PPP rPaPrrre gt e EPP rr Rr rer rrr Fe Oe Oe ee ee Seeteeren nae O Ok cau D 6 fe 2 i e r m g e D D z j 124 KTA avant Operating Instructions 29 1015 56 AA 5 Prepare the system for a run 5 8 Prepare for a run at cold temperature 5 8 Prepare for a run at cold temperature Introduction To fit the AKTA avant instrument in a cold cabinet the foldable door and pump cover can be removed For instructions see AKTA avant User Manual When using the instrument in a cold room or cold cabinet make sure to follow the precautions listed in the next topic Precautions concerning runs in a cold temperature NOTICE e Avoid condensation If KTA avant is kept in a cold room cold cabinet or similar keep it switched on in order to avoid conden sation e Av
86. llation 4 2 5 Prepare the pump rinsing system Step Action 6 Connect a 25 to 30 ml syringe to the outlet tubing of the system pump piston rinsing system Draw liquid slowly into the syringe 7 Disconnect the syringe and discard its contents 8 Immerse the outlet tubing in the rinsing solution tube where the inlet tubing of the system pump piston rinsing system is immersed 9 Connect a 25 to 30 ml syringe to the outlet tubing from the sample pump piston rinsing system Draw liquid slowly into the syringe 10 Disconnect the syringe and discard its contents 11 Immerse the outlet tubing in the rinsing solution tube where the inlet tubing of the sample pump piston rinsing system is immersed 12 Fill the rinsing solution tubes so that each of the tubes contains 50 ml of 20 ethanol 76 KTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 2 Hardware installation 4 2 6 Start the instrument and the computer 4 2 6 Start the instrument and the computer Introduction This section describes how to start the instrument and the computer Instruction Follow the instructions to start the instrument and the computer Step Action 1 Switch on the instrument by pressing the Power switch to the I position Result The instrument starts and the Instrument display states Not connect ed 2 Turn on the computer and monitor according to the manufacturer s instruc tions AKTA avant Operating Instructions 29
87. lways be easy to disconnect e Supply voltage Before connecting the power cord make sure that the supply voltage at the wall outlet corresponds to the marking on the instrument Requirements The following table specifies the power requirements Parameter Requirement Supply voltage 100 240 VAC Frequency 50 60 Hz Transient level Overvoltage category Max power consump 800 VA tion Number of sockets 1 socket per instrument up to 3 sockets for computer equipment Type of sockets EU or US plugs Grounded mains sockets fused or protect ed by equivalent circuit breaker Location of sockets Maximum 2 m from the instrument due to length of mains cable Extension cables can be used if required AKTA avant Operating Instructions 29 1015 56 AA 57 4 Installation 4 1 Site preparation 4 1 4 Power requirements Quality of power The mains power supply must be stable and conform to specifications at all times to ensure reliable operation of the AKTA avant instrument There should be no transient or Slow changes in average voltage outside the limits specified above 58 AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 1 Site preparation 4 1 5 Computer requirements 4 1 5 Computer requirements Introduction AKTA avant systems are controlled by UNICORN software running on a PC The PC can be part of the delivery or be supplied locally The PC used must fulfill the recommen
88. maintenance of the product Immerse the valves completely in methanol and place them in an ultrasonic bath for a few minutes Repeat the ultrasonic bath with deionized water 9 Refit the check valves 10 Tighten the nut until fully finger tight and then use the adjustable wrench to tighten a further 90 degrees 11 Refit the inlet manifold and reconnect the tubing to the pump head 172 AKTA avant Operating Instructions 29 1015 56 AA 8 Reference Information 8 Reference Information About this chapter This chapter lists the technical specifications of the AKTA avant instrument The chapter also includes a chemical resistance guide See also AKTA avant 25 Product Documentation and AKTA avant 150 Product Documentation for detailed technical specifications In this chapter This chapter contains the following sections Section See page 8 1 System specifications 174 8 2 Chemical resistance guide 176 8 3 Check and change the Node ID of a module 181 AKTA avant Operating Instructions 29 1015 56 AA 173 8 Reference Information 8 1 System specifications 8 1 System specifications Technical specifications Parameter System configuration Benchtop system external computer Control system UNICORN 6 0 or later version Connection between PC and instrument Ethernet Dimensions Length x Depth x Height 860 x 710 x 660 mm Weight excluding computer 116 kg Power supply 100 C to 240 C VAC
89. ml Startsystem wash e Set the System flow to 1 0 ml min for KTA avant 25 or 5 0 ml min for AKTA avant 150 e Click Set flow rate Result Only System Pump B is active and a system flow through injection valve waste starts 86 AKTA avant Operating Instructions 29 1015 56 AA Step 4 Installation 4 5 Prime inlets and purge pump heads 4 5 1 Prime buffer inlets and purge system pumps Action 9 Connect a 25 to 30 ml syringe to the purge valve of the left pump head of System Pump B Make sure that the syringe fits tightly into the purge con nector Open the purge valve by turning it counter clockwise about three quarters of a turn Draw 5 to 10 ml of liquid slowly into the syringe with a rate of about 1 ml s Close the purge valve by turning it clockwise Disconnect the syringe and discard its contents Connect the syringe to the purge valve on the right pump head of System Pump B and repeat steps 6 to 8 Keep the system flow running Check that there is no air left in the pump by following the instructions in Validate prime or purge of System Pump A or B or Sample Pump on page 89 AKTA avant Operating Instructions 29 1015 56 AA 87 4 Installation 4 5 Prime inlets and purge pump heads 4 5 1 Prime buffer inlets and purge system pumps Purge System Pump A 88 Purge both pump heads of System Pump A by following the same procedure as in Purge System Pump B on page 85 but replace step 3 a
90. n e Select the position of the inlet to be filled Start at the inlet position with the highest number and end at the position with the lowest number or the buffer position assuming that the first sample to run is connected to inlet 1 etc Buf Sample inlet valve Ls Butter 1 S2 3 S4 s5 fess asz Result The sample inlet valve switches to the selected port Connect a 25 to 30 ml syringe to one of the purge valves of the pump heads in the sample pump Make sure that the syringe fits tightly into the purge connector 6 Open the purge valve by turning it counter clockwise about three quarters of a turn Draw slowly with the syringe until the sample just passes the Sample inlet valve 7 Close the purge valve by turning it clockwise Disconnect the syringe and discard its contents 8 Repeat steps 2 to 5 for each sample inlet that is to be used in the method run The final sample or the buffer from the buffer position should be drawn all the way through both pump heads into the syringe AKTA avant Operating Instructions 29 1015 56 AA 91 4 Installation 4 5 Prime inlets and purge pump heads 4 5 2 Prime sample inlets and purge Sample Pump Step Action 9 Check that there is no air left in the pump by following the instructions in Validate prime or purge of System Pump A or B or Sample Pump on page 89 If air bubbles are indicated follow the instructions in Purge Sample Pump on page 92 Purge Sample Pum
91. n regarding recycling are also described Important WARNING Before installing operating or maintaining the product all users must read and understand the entire contents of this chapter to become aware of the hazards involved In this chapter Section See page 2 1 Safety Precautions 15 2 2 Labels 24 2 3 Emergency procedures 27 2 4 Recycling information 30 2 5 Declaration of Hazardous Substances DoHS 31 14 KTA avant Operating Instructions 29 1015 56 AA 2 Safety Instructions 2 1 Safety Precautions 2 1 Safety Precautions Introduction The safety precautions in this section are grouped in the following categories General precautions on page 15 Flammable liquids and explosive environment on page 16 Personal protection on page 17 Installing and moving on page 18 System operation on page 20 Maintenance on page 22 General precautions WARNING Risk assessment Perform a risk assessment for any risks due to the process or process environment Evaluate the effects the use of the product and the operational processes may have on the classification of the hazardous area The process might cause the area to increase or the zone classification to change Implement the risk reduction measures needed including use of personal protective equipment WARNING Always follow these General precautions to avoid injury when using the AKTA avant instrument e Do not operate t
92. nd 4 with the following Step Action 3 In the Process Picture e Click the Inlet valve A icon e Click the position of one of the inlets that will be used at the beginning of the run aj Inlet vake A at Ad a2 Azlan asl gel az Result The inlet valve switches to the selected port 4 In the Process Picture e Click the System pumps icon e Set Conc B to 0 B and click Set B System flow 4 1 000 mi min Set flow rate Conc B 0 0 B Set B Pump wash A B Al v Start pump A wash B1 v Start pump B wash Q1 v Start Q inlet wash System wash System out W Injection valve W1 15 m Start system wash 10 999 Result Only System Pump A is active AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 5 Prime inlets and purge pump heads 4 5 1 Prime buffer inlets and purge system pumps Validate prime or purge of System Pump A or B or Sample Pump Follow these instructions to check that there is no air left in the pump after performing a prime or a purge Step Action il In the Process Picture e Click on the Injection valve and select Manual load Result The injection valve switches to manual load position Manual load System pump waste Inject Sample pump load Direct inject Sample pump waste 2 Make sure that the pump flow is on 3 In the Chromatogram pane e Check the PreC pressure curve e lf the PreC pressure do not stabilize within a few minutes the
93. nect the system units Connect to network Follow the instructions to make network connections Step Action 1 Connect a network cable between the network connector Ethernet on the back of the instrument and the computer network card dedicated to AKTA The illustration shows the symbol of the Ethernet connector g If the computer is to be connected to an external network connect a network cable between the main network card of the computer and a network wall outlet Note If the computer has not been supplied by GE and if network configuration is to be used see UNICORN Administration and Technical Manual for further information on network settings AKTA avant Operating Instructions 29 1015 56 AA 69 4 Installation 4 2 Hardware installation 4 2 3 Prepare waste tubing 4 2 3 Prepare waste tubing Location of waste tubing All waste tubing is found on the rear of the instrument see the following illustration i eee ee ee aes a a 2 222 S2EE886EE6E2668E 568 Se cveeneenoaaeon nen 2eqaeeaesneeaen Bee Description Waste tubing from the injection valve the pH valve and the outlet valve pieces of tubing marked W W1 W2 and W3 Waste tubing from the fraction collector and the buffer tray 70 AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 2 Hardware installation 4 2 3 Prepare waste tubing Prepare the waste tubing Follow the instructions to prepare the waste tubi
94. ng Step Action 1 Place the four pieces of waste tubing from the injection valve the pH valve and the outlet valve pieces of tubing marked W W1 W2 and W3 ina vessel placed below the bench NOTICE The maximum level of the waste vessel for the waste tubing from the valves must be lower than 30 cm above the lab bench Place the three pieces of waste tubing from the fraction collector and the buffer tray in a waste vessel placed below the bench NOTICE The maximum level of the waste vessel for the waste tubing from the fraction collector and the buffer tray must be lower than the bench height Cut the waste tubing from the fraction collector and the buffer tray to appro priate length It is important that the tubing is not bent and will not be sub merged in liquid during the run q KTA avant Operating Instructions 29 1015 56 AA 71 4 Installation 4 2 Hardware installation 4 2 3 Prepare waste tubing Note If the tubing is too short replace it with new tubing Do not lengthen the tubing as this might cause obstruction of the tubing and flooding in the fraction col lector chamber CAUTION Make sure that the waste vessels will hold all the produced volume of the run For AKTA avant 25 a suitable waste vessel should typi cally have a volume of 2 to 10 liters For AKTA avant 150 a waste vessel should have a volume of 40 liters 72 AKTA avant Operating
95. ng sections Section See page 6 1 Before you start 127 6 2 Applying the sample 130 6 3 Start a method run 133 6 4 Monitor the run 139 6 5 After run procedures 142 126 KTA avant Operating Instructions 29 1015 56 AA 6 Runa method 6 1 Before you start 6 1 Before you start Introduction Before starting a run it is necessary to read and understand the information in this section and to perform the checks listed in the next topic WARNING e Always use appropriate Personal Protective Equipment PPE during operation and maintenance of this product e Hazardous substances When using hazardous chemicals take all suitable protective measures such as wearing protec tive glasses and gloves resistant to the substances used Follow local and or national regulations for safe operation and maintenance of the product e High pressure The product operates under high pressure Wear protective glasses and other required Personal Protective Equipment PPE at all times Checklist Make sure that the system is correctly prepared Prepared the system according to the settings in the method to be run Select a suitable column for the application Immerse the buffer inlet tubing in correct buffer vessels Immerse all waste tubing in appropriate waste vessels consider vessel size placement and material Make sure no tubing is twisted and that the flow path is free from leakage AKTA avant Operating Instru
96. ng showed that there was no air left in the pump it is not necessary to purge Quaternary Valve and the system pumps However if the pressure signal indicated air bubbles left in the valve or the pump follow these instructions to purge Quaternary Valve System Pump A and System Pump B Note that both pump heads of each system pump have to be purged AKTA avant Operating Instructions 29 1015 56 AA 97 4 Installation 4 5 Prime inlets and purge pump heads 4 5 3 Prime Q inlets Step Action l In the Manual instructions dialog box e Select Pumps and pressures Pump wash and click All on the BufferPro Q inlets menu Manual instructions System3 Selected column type __ Select Instructions p Parameters for Pump wash Pumps and pressures n Inlet amp System flow Sample flow Ott Mi Gradient Inlet B Pump wash System wash orf he Quaternary start concentrations BufferPro Q inlets Quaternary gradient BufferPro pH x All Mi Column packing flow Sample inlet Flow path Monitors a Off be Save result as Browse Auto update of parameters during run e Click Execute Result A simultaneous pump wash of all the Q inlets is started This will re move air from Quaternary Valve 2 Wait until the pump wash is completed 3 Select Pumps and pressures System flow and set Flow rate to 0 01 ml min 4 Connecta 25 to 30 ml syringe to the left purge valve o
97. ng the instrument All lifting and moving must be performed in accordance with local regulations e Moving the product horizontally Three people are required to move the product horizontally e Supply voltage Before connecting the power cord make sure that the supply voltage at the wall outlet corresponds to the marking on the instrument e Protective ground The product must always be connected to a grounded power outlet e Power cord Only use power cords with approved plugs deliv ered or approved by GE 18 AKTA avant Operating Instructions 29 1015 56 AA 2 Safety Instructions 2 1 Safety Precautions WARNING Access to power switch and power cord with plug Do not block access to the power switch and power cord The power switch must always be easy to access The power cord with plug must always be easy to disconnect Installing the computer The computer must be installed and used according to the instructions provided by the manufac turer of the computer AKTA avant Operating Instructions 29 1015 56 AA NOTICE To avoid damage to the instrument when installing and moving the KTA avant system take the following measures Make sure that the waste vessels will hold all the produced volume of the run For AKTA avant 25 a suitable waste vessel should typically have a volume of 2 to 10 liters For AKTA avant 150 a waste vessel should have a volume of 40 liters The maximum level of the
98. nsing systems The following illustration shows the tubing configuration of the pump piston rinsing systems Part Description Inlet tubing to the sample pump piston rinsing system Outlet tubing from the sample pump piston rinsing system Inlet tubing to the system pump piston rinsing system Outlet tubing from the system pump piston rinsing system 74 KTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 2 Hardware installation 4 2 5 Prepare the pump rinsing system Prime the pump piston rinsing system Follow the instructions to fill the pump piston rinsing systems with rinsing solution See the tubing configuration of the rinsing systems in Illustration of the pump piston rinsing systems on page 74 Step Action 1 Unscrew the rinsing system tubes from the holders 2 Fill each of the rinsing system tubes with 50 ml of 20 ethanol 3 Screw the rinsing solution tubes back in the holders 4 Immerse the inlet tubing to the system pump piston rinsing system in one of the rinsing solution tubes Note Make sure that the inlet tubing reaches close to the bottom of the rinsing so lution tube 5 Immerse the inlet tubing to the sample pump piston rinsing system in the other rinsing solution tube Note Make sure that the inlet tubing reaches close to the bottom of the rinsing so lution tube KTA avant Operating Instructions 29 1015 56 AA 75 4 Installation 4 2 Hardware insta
99. nt Room climate The following requirements must be fulfilled e The instrument is intended for indoor use only e The room must have exhaust ventilation e The instrument should not be exposed to direct sunlight e Dustin the atmosphere should be kept to a minimum Allowed temperature and humidity ranges are specified in the following table Parameter Ambient temperature operating Allowed range 4 C to 35 C Ambient temperature storage 25 C to 60 C Relative humidity operating 20 to 95 noncondensing Altitude Maximum 2000 m Pollution degree Heat output 2 The heat output data is listed in the following table Component AKTA avant instrument Heat output 800 W Computer incl monitor and printer Typically 300 W Total heat output 1100 W AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 1 Site preparation 4 1 4 Power requirements 4 1 4 Power requirements Introduction This section describes the power supply requirements for the AKTA avant instrument WARNING e Protective ground The product must always be connected to a grounded power outlet e Power cord Only use power cords with approved plugs deliv ered or approved by GE e Access to power switch and power cord with plug Do not block access to the power switch and power cord The power switch must always be easy to access The power cord with plug must a
100. of the column valve e g ports 1A and 1B e For Pump P9H Connect the reference capillary Ref 1 or an equivalent capillary that gives a backpressure of 2 to 3 MPa 20 to 30 bar to one of the column positions of the column valve e g ports 1A and 1B 4 Immerse the waste tubing in the buffer vessel to recirculate the liquid 5 e fbreaking in a pump piston seal of a system pump follow the instruction below In the Manual instructions dialog box Select Flow path Column valve and select the position of the capil lary connected to the column valve Click Insert Select Flow path Inlet A for System Pump A or Flow path Inlet B for System Pump B and select a Position Click Insert Select Pumps and Pressures Gradient and set Target to 0 B for System Pump A or 100 B for system pump B For Pump P9 Select Pumps and Pressures System flow and set the Flow rate to 5 0 to ml min Click Insert For Pump P9H Select Pumps and Pressures System flow and set the Flow rate to 25 0 to ml min Click Insert Click Execute Result A system flow starts e If breaking ina pump piston seal of a sample pump follow the instruction below In the Manual instructions dialog box Select Flow path Column valve and select the position of the capil lary connected to the column valve Click Insert Select Flow path Sample inlet and select a Position Click Insert Select Flow path Injection valve and click Direct inject on the Posi
101. oid overheating If KTA avant is kept in a cold cabinet and the cold cabinet is switched off make sure to switch off AKTA avant and keep the cold cabinet open to avoid overheating e Place the computer in room temperature If the KTA avant instrument is placed in a cold room use a cold room compat ible computer or place the computer outside the cold room and use the Ethernet cable delivered with the instrument to connect to the computer Note When the instrument is kept in a cold room it is important to tighten all tubing connectors also the inlet manifold connectors Otherwise air might get into the flow path Note Make sure that the instrument buffers and sample have had time to reach the ambient temperature When the instrument has reached the ambient temperature calibrate all pressure sensors Tip When runs are performed in a cold cabinet make sure to adjust the target temperature of the built in fraction collector temperature control function The target temperature is 20 C by default Settings for the temperature control function can be edited in the System Settings dialog box of System Control or in the Text Instructions pane in Method Editor AKTA avant Operating Instructions 29 1015 56 AA 125 6 Runamethod 6 Runa method About this chapter This chapter describes how to start up and run a method and also how to handle the system after the run In this chapter This chapter contains the followi
102. oided These might cause swelling of plastic material and reduce the pressure tolerance of PEEK tubing For this reason flash chromatography and straight normal phase chromatography is generally not recommended on the system KTA avant Operating Instructions 29 1015 56 AA 8 Reference Information 8 2 Chemical resistance guide Assumptions made The ratings are based on the following assumptions e Synergy effects of chemical mixtures have not been taken into account e Room temperature and limited overpressure is assumed Note Chemical influences are time and pressure dependent Unless otherwise stated all concentrations are 100 List of chemicals Note A user can be exposed to large volumes of chemical substances over a long time period Material Safety Data Sheets MSDS provide the user with informa tion regarding characteristics human and environmental risks and preventive measures Make sure that you have the MSDS available from your chemical distributor and or databases on the internet Aqueous buffers Continuous use Chemical Concentra CAS no EC no tion Aqueous buffers N A N A pH 2 12 Strong chemicals and salts for CIP Up to 2 h contact time at room temperature Chemical Concentra CAS no EC no tion Acetic acid 75 05 8 200 835 2 Decon 90 N A Ethanol 75 08 1 200 837 3 Methanol 100 67 56 1 200 659 6 Hydrochloric acid 1 0 1M 7647 01 0 231 595 7 lsopropanol
103. ollowing table If you want to pause the run e Press the Pause button on the instrument display This will stop all pumps in the instrument AKTAavani or e To pause the run from UNICORN click the Pause button in the System Control module tt System Control File Edit view Manual System Tools Help Result All pumps in the instrument are stopped AKTA avant Operating Instructions 29 1015 56 AA 27 2 Safety Instructions 2 3 Emergency procedures If you want to switch offthe Push the Power switch to the O position instrument or e disconnect the power cord from the wall socket Result The run is interrupted immediately Note The sample and data may be lost as a result of switching off the power Power failure The result of a power failure depends on which unit is affected Power failure to will result in AKTA avant instrument The run is interrupted immediately The data collected up to the time of the power failure is available in UNICORN Computer The UNICORN computer shuts down The instrument display shows status Not connected The run is interrupted immediately Data generated up to 10 seconds before the power failure can be recovered Note The UNICORN client may lose connection to the instrument during a temporary overload of the processor and display an error message This may appear as a computer failure The run continues
104. or ex ample a held or paused method run End Permanently ends the method run Customize Opens the Cus tomize dialog box where curve settings run data groups and run log con tents can be set KTA avant Operating Instructions 29 1015 56 AA Connect to Systems Opens the Connect to Systems di alog box where systems can be connected and cur rently connected users are displayed 47 4 Installation 4 Installation About this section This section provides the instructions necessary to enable users and service personnel to install the instrument install the computer and install the software Read the entire Installation chapter before starting to install the AKTA avant instrument Note For information on how to how to unpack the AKTA avant instrument and how to lift the instrument onto a laboratory bench see AKTA avant Unpacking In Structions In this section This sections contains the following subsections Section See page 4 1 Site preparation 49 4 2 Hardware installation 64 4 3 Software installation 78 4 4 Start UNICORN and connect to system 19 4 5 Prime inlets and purge pump heads 82 4 6 Performance tests 100 48 AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 1 Site preparation 4 1 Site preparation Introduction This subsection describes the site planning and the preparations necessary to perform before installation of an AKTA avant system The purp
105. ose is to provide planners and technical staff with the data needed to prepare the laboratory for the installation The laboratory site must be planned and prepared before installing the AKTA avant system The performance specifications of the system can be met only if the laboratory environment fulfills the requirements stated in this chapter The time spent in preparing the laboratory will contribute to the long term performance of the systems In this subsection Section See page 4 1 1 Delivery and storage 50 4 1 2 Room requirements 52 4 1 3 Site environment 56 4 1 4 Power requirements 57 4 1 5 Computer requirements 59 4 1 6 Required materials 61 KTA avant Operating Instructions 29 1015 56 AA 49 4 Installation 4 1 Site preparation 4 1 1 Delivery and storage 4 1 1 Delivery and storage Introduction This section describes the requirements for receiving the delivery box and storing the instrument before installation WARNING Heavy object The AKTA avant instrument weighs about 116 kg Use proper lifting equipment or use four or more persons when moving the instrument All lifting and moving must be performed in accordance with local regulations When you receive the delivery e Record on the receiving documents if there is any apparent damage on the delivery box Inform your GE representative of such damage e Move the delivery box to a protected location indoors Delivery box AKTA avant ins
106. p If the priming was done thoroughly and the final buffer was drawn all the way into the syringe and the validation of the priming showed that there was no air left in the pump it is not necessary to purge the sample pump However if the pressure signal indicated air bubbles left in the pump follow the instruction below to purge both the pump heads of the sample pump Step Action 1 Make sure that all sample inlet tubing that is to be used during the method run is immersed in the correct buffers 2 Make sure that the waste tubing connected to injection valve port W2 is immersed in a waste vessel 3 Open the System Control module 4 In the Process Picture e Click the Injection valve icon and then click Sample pump waste amp E w1 Co POA op SyP F I en Injection valve x Manual load System pump waste Inject Sample pump load Direct inject Sample pump waste Result The injection valve switches to waste position This is necessary to achieve a low back pressure during the purge procedure 92 AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 5 Prime inlets and purge pump heads 4 5 2 Prime sample inlets and purge Sample Pump Step Action 5 In the Process Picture e Click the Sample inlet icon then click Buffer e Click the Sample pump icon Set the Sample flow to 1 0 ml min for AKTA avant 25 or 5 0 ml min for AKTA avant 150 Inlets Pumps Sample De pump waste ry gt ei
107. r Do not fractionate flammable liquids in the built in fraction collector When running RPC methods collect fractions through the outlet valve or the optional external Fraction Collector F9 R 16 AKTA avant Operating Instructions 29 1015 56 AA 2 Safety Instructions 2 1 Safety Precautions WARNING e RPC runs with 100 acetonitrile and system pressure above 5 MPa 50 bar in AKTA avant 25 Always replace the green PEEK tubing between the used system pump and the pump pressure monitor with orange PEEK tubing i d 0 5 mm before running RPC with 100 acetonitrile Set the system pressure alarm to 10 MPa 100 bar e RPCruns with 100 acetonitrile in KTA avant 150 Always replace the beige PEEK tubing between the used system pump and the pump pressure monitor before running RPC with 100 acetonitrile Replace it with green PEEK tubing i d 0 75 mm Personal protection WARNING To avoid injury when working with the AKTA avant system take the following measures for personal protection e Always use appropriate Personal Protective Equipment PPE during operation and maintenance of this product e Hazardous substances and biological agents 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 AKTA avant
108. rations The illustrations below show the parts of the pump heads of the pumps P9 and P9H Pump P9 150 AKTA avant Operating Instructions 29 1015 56 AA 7 Maintenance 7 2 Replace pump piston seal O rings and rinse membrane of Pump P9 or Pump P9H Pump P9H Description Piston Description Piston seal Return spring Outlet check valve Pump membrane housing Star screws O ring Purge valve O ring Star screws Support washer Inlet check valve Star screws Disassemble the pump head Step Action 1 Make sure that no run is in progress on the instrument 2 Disconnect the tubing from the pump head and disconnect the pump inlet tubing AKTA avant Operating Instructions 29 1015 56 AA 151 7 Maintenance 7 2 Replace pump piston seal O rings and rinse membrane of Pump P9 or Pump P9H Step Action 3 Unscrew the two white plastic screws located below each pump head by hand Pull the plastic connectors to the sides to release the inlet manifold KS ds 1 4 Disconnect the tubing of the pump piston rinsing system 5 Unscrew the two screws of the front section of the pump head using a star screwdriver T20 and pull off the front section 152 AKTA avant Operating Instructions 29 1015 56 AA 7 Maintenance 7 2 Replace pump piston seal O rings and rinse membrane of Pump P9 or Pump P9H Step Action 6 Place the front section of the pump head face down on the
109. re may be air left in the pump See AKTA avant User Manual End the run Click the End button in the System Control toolbar to end the run AKTA avant Operating Instructions 29 1015 56 AA 89 4 Installation 4 5 Prime inlets and purge pump heads 4 5 2 Prime sample inlets and purge Sample Pump 4 5 2 Prime sample inlets and purge Sample Pump Overview The procedure consists of the following stages Stage 1 2 Description Prime all sample inlet tubing to be used during the run Validate priming of inlet tubing Purge the sample pump if pressure signal indicates air bubbles Validate purge of the samle pump End the run Note To increase life length of the pump sealing rings make sure that the pump rinsing system is filled with fresh rinsing solution Prime sample inlets Follow the instructions below to fill all sample inlet tubing to be used in the run with appropriate buffer or sample solution Step Action 1 90 Make sure that all sample inlet tubing that is to be used during the method run is immersed in the correct samples Make sure that the waste tubing connected to injection valve port W2 is immersed in a waste vessel Open the System Control module AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 5 Prime inlets and purge pump heads 4 5 2 Prime sample inlets and purge Sample Pump Step Action 4 In the Process Picture e Click the Sample inlet valve ico
110. red operation Note The user is cautioned that any changes or modifications not expressly approved by GE could void the user s authority to operate the equipment This equipment has been tested and found to comply with the limits for a Class A digital device pursuant to part 15 of the FCC Rules These limits are designed to provide rea sonable protection against harmful interference when the equipment is operated in a commercial environment This equipment generates uses and can radiate radio frequen cy energy and if not installed and used in accordance with the instruction manual may cause harmful interference to radio communications Operation of this equipment in a residential area is likely to cause harmful interference in which case the user will be re quired to correct the interference at his own expense Environmental conformity 10 This product conforms to the following environmental requirements Requirement 2011 65 EU Restriction of Hazardous Substances RoHS Directive 2012 19 EU Waste Electrical and Electronic Equipment WEEE Directive ACPEIP Administration on the Control of Pollution Caused by Electronic Information Products China Restriction of Hazardous Sub stances RoHS Regulation EC Registration Evaluation Authorization and restriction of No 1907 2006 CHemicals REACH AKTA avant Operating Instructions 29 1015 56 AA 1 Introduction 1 3 Regulatory information Regulatory comp
111. rm can be set in the method to be run the System Settings dialog box or during a manual run Precolumn pressure alarm limits are automatically set in the method when a column from the column list is selected in the method Refer to UNICORN Method Manual for more information on pressure alarms Set pressure alarms Pressure alarm limits may be set manually in System Control The example below de scribes how to set the high pressure limit for the column Other alarms are set in a cor responding way Step Action 1 In the System Control module on the Manual menu click Execute Manual Instructions Result The Manual instructions dialog box opens 2 In the Instructions box select Alarms Alarm pre column pressure Manual instructions System 1 Selected column type Parameters for Slarm pre column pressure Mode Instructions Instruction execution list Pumps Flow path S Honiara O Disabled Enabled Fraction collection High alarm 0 02 20 00 S Alarms Alarm system pressure 0 50 MPa Alarm pre column pressure Low alarm 10 00 20 00 Alarm UY z Alarm conductivity 0 30 gt MPa Wash settings Watch parameters Advanced v Save result as L_ Browse V Auto update of parameters during run 3 Click Enabled in the Mode field 4 e Enter the high pressure limit in the High alarm box e Click Execute 116 AKTA avant Operating Instructions 29 1015 56 AA 5
112. ruc tions e 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 dam age to the flow cell may occur e Glass tube splinter Make sure to set the sample pressure below the maximum pressure of the Superloop before execut ing a flow in the Manual instructions dialog box when the Superloop is connected e Avoid condensation If KTA avant is kept in a cold room cold cabinet or similar keep it switched on in order to avoid conden sation e Avoid overheating If KTA avant is kept in a cold cabinet and the cold cabinet is switched off make sure to switch off KTA avant and keep the cold cabinet open to avoid overheating e Place the computer in room temperature If the KTA avant instrument is placed in a cold room use a cold room compat ible computer or place the computer outside the cold room and use the Ethernet cable delivered with the instrument to connect to the computer e UV and conductivity flow cells on the high pressure side When placing UV and or conductivity flow cells on the high pressure side of the column the UV flow cell has a maximum pressure limit of 2 MPa 20 bar and the conductivity flow cell has a maximum pressure limit of 5 MPa 50 bar Maintenance WARNING To avoid personal injury when performing maintenance on the KTA avant instrument follow these
113. s Pras ASWANTtANEhMReS Table of Hazardous Substances Name and Concentration bE E ABA SWAN Component name Hazardous substance R 7K tf KER BRERA Hg Cd Cr6 PBB 28 9308 42 O O O 28 9763 37 O O O 29 0113 60 O O O 29 0113 61 O O O 29 0113 62 O O O 29 0906 89 O O O 29 0906 91 O O O 29 0113 53 O O O 29 0113 58 O O O 0 RRZASAS MAE Aa MAMA YS B55 ESJ T11363 2006 VEAN TE AY BR BE KAP x RRZASASVRSY EZRA YRMS HA BBB wSJ 111363 2006 ie 7E BY BR ee BE SR e We Fe PIT ON BHA Ac Hh BY PT ESA 1S BY Be 1S 0 Indicates that this toxic or hazardous substance contained in all of the homogeneous ma terials for this part is below the limit requirement in SJ T11363 2006 X Indicates that this toxic or hazardous substance contained in at least one of the homoge neous materials used for this part is above the limit requirement in SJ T11363 2006 Data listed in the table represents best information available at the time of publication 32 KTA avant Operating Instructions 29 1015 56 AA 3 System description 3 System description About this chapter This chapter gives an overview of the AKTA avant instrument software and accessories In this chapter This chapter contains the following sections Section See page 3 1 AKTA avant instrument overview 34 3 2 UNICORN software 43 Illustration of the system The following illustration shows the AKTA avant instrument with UNICORN sof
114. s chapter This chapter contains the following sections Section See page 5 1 Before you prepare the system 102 5 2 Prepare the flowpath 104 5 3 Prime buffer inlets and purge system pumps 109 5 4 Connect a column 110 5 5 Set pressure alarms 115 5 6 Calibrate the pH monitor 117 5 7 Prepare the built in fraction collector 119 5 8 Prepare for a run at cold temperature 125 101 AKTA avant Operating Instructions 29 1015 56 AA 5 Prepare the system for a run 5 1 Before you prepare the system 5 1 Before you prepare the system Introduction It is important to prepare the system in accordance with the settings in the method to be run Before preparing the system check the settings in the Method Editor and make sure that all accessories to be used are available WARNING e Do not use KTA avant if it is not working properly or if it has suffered any damage for example damage to the power cord or its plug damage caused by dropping the equipment damage caused by splashing liquid onto it e Always use appropriate Personal Protective Equipment PPE during operation and maintenance of this product e Do not use any accessories not supplied or recommended by GE e Fire Hazard Before starting the system make sure that there is no leakage Checklist Remember to check the following 102 which valve ports to use for inlets and outlets which column type to use which column position to use which
115. s items in a group thus Flowpath Injection valve refers to the Injection valve item in the Flowpath group Hardware items are identified in the text by bold text for example the Power button AKTA avant Operating Instructions 29 1015 56 AA 5 1 Introduction 1 2 Important user information 1 2 Important user information Read this before operating the product All users must read the entire Operating Instructions before installing operating or maintaining the product Always keep the Operating Instructions at hand when operating the product Do not operate the product 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 of the product KTA avant is a liquid chromatography system intended for method and process devel opment in purification of biomolecules The system can be used to screen for optimal choice of columns media and running parameters to purify selected proteins The AKTA avant system is intended for research use only and shall not be used in any clinical procedures or for diagnostic procedures Prerequisites In order to follow this manual and use the system in the manner it is intended it is im portant that e You have a general understanding of how the computer and Microsoft Windows work e You understand the concepts of liquid chromatography e You h
116. sing solution volume starts to increase NOTICE e Do not disassemble the pump head unless there is good reason to believe that there is an internal leakage A sign of leakage is increasing pump rinsing solution volume Always make sure that sufficient spare components are available before attempt ing to replace a spare part e Replacing spare parts Read the instructions carefully For example some individual parts of the pump head can be as sembled incorrectly Check the orientation of each part before continuing with the next instruction Maintenance interval Replace the O rings piston seals and rinse membranes the pumps P9 P9H A P9H B and P9H S if they are damaged After replacement perform a run to break in the new piston seals NOTICE Advanced maintenance Read the instruction carefully before disassembly of the pump head KTA avant Operating Instructions 29 1015 56 AA 149 7 Maintenance 7 2 Replace pump piston seal O rings and rinse membrane of Pump P9 or Pump P9H Required material The following materials are required e Adjustable wrench e For Pump P9 Star screwdriver T20 e For Pump P9H Star screwdriver T10 and T20 e Ultrasonic bath e Ethanol 20 e For Pump P9 Tubing giving a back pressure of 6 to 8 MPa 60 to 80 bar e For Pump P9H Tubing giving a back pressure of 2 to 3 MPa 20 to 30 bar e For Pump P9 P9 Seal kit 25 ml e For Pump P9H P9H Seal kit 150 ml Illust
117. ston seal O rings and rinse membrane of Pump P9 or Pump P9H Replace O rings piston seal and pump membrane housing Step Action 1 Unscrew the purge valve of the pump head Replace the O ring on the purge valve with a new O ring and screw the purge valve back into the pump head Note Always use Lubricant 56686700 when exchanging the O ring 3 x 1 mm 2 Wet a new seal with 20 ethanol Place the new seal in the hole in the front section of the pump head and press it into position Pump P9 Pump P9H AKTA avant Operating Instructions 29 1015 56 AA 155 7 Maintenance 7 2 Replace pump piston seal O rings and rinse membrane of Pump P9 or Pump P9H Step Action 3 Place the support washer on top of the new seal in the front section of the pump head Screw the two or four screws of the support washer Make sure to tighten the screws fully Pump P9H 4 Wet a new O ring 21 4 x 1 6 mm with 20 ethanol Fit the O ring around the support washer Pump P9 Pump P9H AKTA avant Operating Instructions 29 1015 56 AA 7 Maintenance 7 2 Replace pump piston seal O rings and rinse membrane of Pump P9 or Pump P9H Assemble the pump head Make sure to assemble the pump head correctly Refer to Illustrations on page 150 Step Action 1 Insert the piston into the return spring Insert piston and return spring into hole in the pump module Note Do no touch the ceramic or glass part of the pump piston 2 Wet the
118. sult The first New Column dialog box opens New Column Code lot Exp ID C The Column has a uniT ag has a fixed Code and exp AKTA avant Operating Instructions 29 1015 56 AA 135 6 Runa method 6 3 Start a method run Step Action 3 Register the column using the Barcode Scanner 2 D as follows e Make sure that the pointer is placed in the first position of the Code box e Point the Barcode Scanner 2 D towards the data matrix tag on the col umn e Press and hold the trigger to create a beam e When the scanner beeps the column ID Is registered and displayed in the dialog box e Alternatively manually enter the column ID that you find on the column label in the dialog box using your keyboard e Click Continue Result The expanded New Column dialog box opens New Column Column ID Alias optional Technique Desatting Column type HiPrep 26 10 Desalting C Use medium batch ID 136 AKTA avant Operating Instructions 29 1015 56 AA 6 Runa method 6 3 Start a method run Step Action 4 In the expanded New Column dialog box e Enter an column alias in the Alias box optional e Click a chromatography technique on the Technique menu e Click a column type on the Column type menu e Select the Set medium expiration date check box and click a date on the menu Tip Alias can be used for easy identification of a column Result The entered
119. t the instrument and the computer 77 64 AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 2 Hardware installation 4 2 1 Install the computer equipment 4 2 1 Install the computer equipment Introduction The computer is supplied as a part of the AKTA avant delivery or supplied locally Unpacking and installing Unpack and install the computer according to the manufacturer s instructions NOTICE Any computer used with the equipment must comply with IEC 60950 and be installed and used according to the manufacturer s instructions KTA avant Operating Instructions 29 1015 56 AA 65 4 Installation 4 2 Hardware installation 4 2 2 Connect the system units 4 2 2 Connect the system units Introduction The following interconnections must be made e power supply to the KTA avant instrument e power supply to the computer equipment e network connection between the computer and the KTA avant instrument WARNING e Power cord Only use power cords with approved plugs deliv ered or approved by GE Supply voltage Before connecting the power cord make sure that the supply voltage at the wall outlet corresponds to the marking on the instrument Illustration The following illustration shows the location of the connectors Function 1 Power input connector 2 Network connector Ethernet 66 KTA avant Operating Instructions 29 1015 56 AA 4 Installa
120. ted onto the column during the run Tip It is recommended to overload the loop to make sure that the loop is com pletely filled Excess of sample will leave the valve through port W1 AKTA avant Operating Instructions 29 1015 56 AA 131 6 Runa method 6 2 Applying the sample Sample application through a sample loop 132 A sample loop is manually filled with sample using a syringe connected to the Injection Valve port Syr During the method run the sample is automatically injected onto the column The loop is emptied and washed out using buffer from the system pumps The total buffer volume to be used for emptying and washing the sample loop is set in the Phase Properties tab of the Sample Application phase in the Empty loop with box Phase Properties Text Instructions T Sample Application Use the same flow rate as in Method Settings Flow rate 10 000 ml min 0 000 25 000 Inject sample from loop Fill the loop using Manual load v l l Loop type Capillary loop v ml Inject sample directly onto column ae Sample inlet Fill loop wi 1 00 ml ml Empty loop with Sample volume C Use the same inlets as in Method Settings Inlet amp Al v InletB BY a oo x _ Fill the system with the selected buffer Tip Empty the sample loop with a buffer volume that exceeds the volume of the loop This will ensure that the loop is comp
121. thod 6 4 Monitor the run Actions in the Process Picture pane It is possible to interact with the Process Picture pane e To open a related instruction click the component icon The example below shows the pop up toolbar for the Injection valve icon Instructions can be given from the pop up toolbar of each component Icon P Injection valve uJ Manual load system pump waste Inject Sample pump load Direct inject Sample pump waste e To display a detailed picture with explanations for example for a valve right click the component icon and click Detailed picture AKTA avant Operating Instructions 29 1015 56 AA 141 6 Runa method 6 5 After run procedures 6 5 After run procedures Introduction This section describes how to clean the instrument and columns after a chromatographic run and how to prepare the system for storage The instrument and the columns should be cleaned between the runs This will prevent for example sample contamination protein precipitation and column clogging If the instrument is not going to be used for a couple of days or longer the instrument columns and the pH flow cell should be filled with storage solution For further information about cleaning and maintenance procedures see Chapter 7 Maintenance on page 145 Tip To clean and fill the instrument and columns with storage solution use the System CIP and Column CIP methods Either as separate predefined methods or as phases incl
122. ting Instructions 29 1015 56 AA 2 Safety Instructions 2 1 Safety Precautions WARNING e Hazardous biological agents during run When using haz ardous biological agents run System CIP and Column CIP to flush the entire pump with bacteriostatic solution e g 1M NaOH followed by a neutral buffer and finally distilled water before service and maintenance CAUTION To avoid hazardous situations when operating the AKTA avant system follow these instructions e Risk of breaking test vials Do not use excessive force to press vials of wrong dimensions into the fraction collector cassettes Glass vials may break and cause injuries e Hazardous chemicals in UV flow cell Make sure that the entire flow cell has been flushed thoroughly with bacteriostatic solu tion for example NaOH and distilled water before service and maintenance e pH electrode Handle the pH electrode with care The glass tip may break and cause injury e Do not fasten bottles with a volume of more than 1 liter in the front panel rails e Max weight on buffer tray Do not place containers with a volume of more than 10 liters each on the buffer tray The total allowed weight on the buffer tray is 40 kg AKTA avant Operating Instructions 29 1015 56 AA 21 2 Safety Instructions 2 1 Safety Precautions NOTICE To avoid damage to the KTA avant instrument or other equipment when operating the instrument follow these inst
123. tion 4 2 Hardware installation 4 2 2 Connect the system units Function UniNet 9 connectors Note Termination plugs must be connected to the connectors that are not in use Other connectors are for use by authorized service engineers only NOTICE Misuse of UniNet 9 connectors The UniNet 9 connectors at the rear panel should not be mistaken for Firewire connectors Do not connect any external equipment to the UniNet 9 connectors Do not disconnect or move the UniNet 9 bus cable AKTA avant Operating Instructions 29 1015 56 AA 67 4 Installation 4 2 Hardware installation 4 2 2 Connect the system units Connect power to the AKTA avant instrument Follow the instructions to connect power to the AKTA avant instrument Step Action 1 Select the correct power cord to be used Each instrument Is delivered with 2 alternative power cords e Power cord with US plug 2 m e Power cord with EU plug 2 m Discard the unused power cord 2 Connect the power cord to the Power input connector on the back of the instrument and to a grounded wall outlet 100 to 240 VAC 50 to 60 Hz 3 Attach the power cord to the rear of the instrument using the cable clip Connect power to computer equipment Follow the manufacturer s instructions to connect power to the computer monitor and local printer if used 68 AKTA avant Operating Instructions 29 1015 56 AA 4 Installation 4 2 Hardware installation 4 2 2 Con
124. truments are shipped in a delivery box with the following dimensions and weight Contents Dimensions mm Weight AKTA avant instrument with ac 1000 x 900 x 800 155 kg cessories width x height x depth Storage requirements The delivery boxes should be stored at a protected place indoors The following storage requirements must be fulfilled for the unopened boxes Parameter Allowed range Ambient temperature storage 25 C to 60 C Relative humidity 20 to 95 noncondensing 50 KTA avant Operating Instructions 29 1015 56 AA Equipment for transportation 4 Installation 4 1 Site preparation 4 1 1 Delivery and storage The following equipment is recommended for handling the delivery boxes Equipment Pallet mover Specifications Suitable for a lightweight pallet 80 x 100 cm Cart for transporting the instru ment to the lab Unpacking the AKTA avant instrument Dimensioned to accommodate the size and weight of the instrument For information on how to how to unpack the AKTA avant instrument and how to lift the instrument onto a laboratory bench see AKTA avant Unpacking Instructions AKTA avant Operating Instructions 29 1015 56 AA 51 4 Installation 4 1 Site preparation 4 1 2 Room requirements 4 1 2 Room requirements Introduction This section describes the requirements for the transportation route and the room where the AKTA avant instrument is placed WARNIN
125. tware in stalled on a computer AKTA avant Operating Instructions 29 1015 56 AA 33 3 System description 3 1 AKTA avant instrument overview 3 1 AKTA avant instrument overview Introduction This section shows an overview of the AKTA avant instrument Technical details about the instrument and the individual modules are found in AKTA avant User Manual Exterior design The AKTA avant instrument has a modular design with all the liquid handling modules placed on the exterior of the instrument Buffer vessels are placed on the buffer tray on top of the instrument An instrument display is located on the front From this side the built in fraction collector is handled as well as the sample The remaining modules are placed on the right hand side of the instrument This side can be covered by a foldable door and a pump cover By rotating the instrument using the swivel foot any side is easily accessed Operating ranges The following table show some of the operational limits of AKTA avant 25 and AKTA avant 150 Parameter Flow rate Limits KTA avant 25 0 001 to 25 ml min Note When running the Column packing flow instruction the maximum flow rate is 50 ml min KTA avant 150 0 01 to 150 ml min Note When running the Column packing flow instruction the maximum flow rate is 300 ml min Max operating pressure 20 MPa 200 bar 5 MPa 50 bar UV monitor wavelength 34 1
126. u try to exit or log off UNICORN you will see a warning Click Yes to save No to exit without saving or Cancel to stay logged on Shut down the instrument Switch off the instrument by pressing the Power switch to the O position 144 AKTA avant Operating Instructions 29 1015 56 AA 7 Maintenance 7 Maintenance About this chapter This chapter provides schedules for preventive maintenance that should be performed by the user of the AKTA avant instrument Regular maintenance is essential for reliable function and results Refer to the AKTA avant User Manual for detailed instructions Pro cedures that require special attention are also described in this chapter WARNING Always use appropriate Personal Protective Equipment PPE during operation and maintenance of this product In this chapter This chapter contains the following sections Section See page 7 1 Maintenance program 146 7 2 Replace pump piston seal O rings and rinse membrane of Pump 149 P9 or Pump P9H 7 3 Replace pump piston seal O ring and rinse membrane of Pump 160 P9 S 7 4 Replace pump pistons 168 7 5 Clean pump head check valves 170 AKTA avant Operating Instructions 29 1015 56 AA 145 7 Maintenance 7 1 Maintenance program 7 1 Maintenance program Introduction An overview of the preventive maintenance to be performed on the AKTA avant instru ment is outlined in the following list See AKTA avant User Manual for
127. uded in a chromatographic method WARNING Corrosive chemicals during maintenance If the system or column is cleaned with a strong base or acid flush with water afterwards and wash with a weak neutral buffer solution in the last step or phase System cleaning After a method run is completed perform the following e Rinse the instrument with one or several cleaning solution s e g NAOH buffer solu tion or distilled water using the System CIP method e lf applicable empty the fraction collector e Clean all spills on the instrument and on the bench using a moist tissue e Empty the waste vessel e Clean the manual injection port of the injection valve see KTA avant User Manual for detailed instructions e f applicable clean the pH electrode manually and make sure to leave it in an appro priate buffer See AKTA avant User Manual for detailed instructions 142 AKTA avant Operating Instructions 29 1015 56 AA 6 Runa method 6 5 After run procedures System storage If the instrument is not going to be used for a couple of days or longer also perform the following e Fill the system and inlets with storage solution e g 20 ethanol using the System CIP method Column cleaning After a method run is completed perform the following e Clean the column with one or several cleaning solution s using the Column CIP method Column storage If the column is not going to be used for a
128. ufacturing information The table below summarizes the required manufacturing information For further infor mation see the EU Declaration of Conformity DoC document Requirement Content Name and address of manufacturer GE Healthcare Bio Sciences AB Bj rkgatan 30 SE 751 84 Uppsala Sweden Conformity with EU Directives This product complies with the European directives listed in the table by fulfilling the corresponding harmonized standards A copy of the EU Declaration of Conformity is included in the documentation package Directive 2006 42 EC Machinery Directive MD 2004 108 EC Electromagnetic Compatibility EMC Directive 2006 95 EC Low Voltage Directive LVD 1999 5 EC Radio Equipment and Telecommunications Terminal Equipment R amp TTE Directive 8 AKTA avant Operating Instructions 29 1015 56 AA 1 Introduction 1 3 Regulatory information CE marking The CE marking and the corresponding EU Declaration of Conformity is valid for the in strument when it is e used asa stand alone unit or e connected to other products recommended or described in the user documentation and e used in the same state as it was delivered from GE except for alterations described in the user documentation International standards This product fulfills the requirements of the following standards Standard Description EN ISO 12100 Safety of machinery General princi EN ISO standar
129. ure signal indicates air bubbles Validate purge of System Pump B Purge System Pump A if pressure signal indicates air bubbles Validate purge of System Pump A End the run Note Tip To increase life length of the pump sealing rings make sure that the pump rinsing system is filled with fresh rinsing solution The procedures for purging the pump heads and priming the inlets using the Process Picture are described in the following topic It is also possible to perform the procedures from the Manual instructions dialog box Prime inlet tubing Follow the instructions to fill all A and B inlet tubing to be used in the run with appropriate buffer solution Step Action 1 Make sure that all inlet tubing that is to be used during the method run is placed in the correct buffer Open the System Control module AKTA avant Operating Instructions 29 1015 56 AA 83 4 Installation 4 5 Prime inlets and purge pump heads 4 5 1 Prime buffer inlets and purge system pumps Step Action 5 84 In the Process Picture e Click the inlet valve icons Click both the Inlet A and Inlet B icons if both inlets are to be primed e Click the position of the inlet to be filled Fill the positions in reverse al ohabetical order and start with the highest number For example if all the seven inlets in Inlet Valve B are to be filled click them in the following order B7 B6 B1 assuming that B1 is the starting buf
130. waste vessel for the waste tubing from the valves must be lower than 30 cm above the lab bench The maximum level of the waste vessel for the waste tubing from the fraction collector and the buffer tray must be lower than the bench height Vents on the AKTA avant instrument To ensure adequate ventilation Keep papers and other objects away from the vents of the instrument Disconnect power To prevent equipment damage always disconnect the power from the product before an instrument module is removed or installed or a cable is connected or disconnected Misuse of UniNet 9 connectors The UniNet 9 connectors at the rear panel should not be mistaken for Firewire connectors Do not connect any external equipment to the UniNet 9 con nectors other than instrument modules designed for AKTA avant See AKTA avant User Manual Do not disconnect or move the UniNet 9 bus cable 19 2 Safety Instructions 2 1 Safety Precautions System operation WARNING To avoid personal injury when operating the AKTA avant system follow these instructions e Rotating the instrument Make sure that there is always at least 20 cm of free space around the AKTA avant instrument to allow for sufficient ventilation and rotation on the swivel foot When rotating the instrument take care not to stretch or Squeeze tubing or cables A disconnected cable may cause power interruption or network interruption Stretched tubing may cause bottles to fall
131. y GE Healthcare UK Limited Amersham Place Little Chalfont Buckinghamshire HP7 9NA UK GE Healthcare Bio Sciences Corp 800 Centennial Avenue PO Box 1327 Piscataway NJ 08855 1327 USA GE Healthcare Japan Corporation Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 169 0073 Japan 29 1015 56 AA 03 2015 a24
132. ycled according to national and local environmental regulations CAUTION Always use appropriate personal protective equipment when decommissioning the equipment Disposal of electrical components Waste comprising electrical and electronic equipment must not be disposed of as un sorted municipal waste and must be collected separately Please contact an authorized representative of the manufacturer for information concerning the decommissioning of equipment 30 AKTA avant Operating Instructions 29 1015 56 AA 2 Safety Instructions 2 5 Declaration of Hazardous Substances DOHS 2 5 Declaration of Hazardous Substances DoHS AB HESJ T11364 2006 8 FERS mR AMER PERM RAK RABIES The following product pollution control information is provided according to SJ T11364 2006 Marking for Control of Pollution caused by Electronic Information Products BSS m RE Blip in Explanation of Pollution Control Label Aine WAT i SAIS I T11363 2006 E FERA Pas SVAN EZR PRENABASUNA MERKATA AA MHRA AAA PmELREAWAE ABASMAFAREREMAE APEA oo RANMA RP BSARNEAR UPGKrSRSHN RR BUA 7A PRUE Pir FR BA AY R52 FB BRR DR oF PER RET E mie FAP eT oS FT PAE AY ASM RAE on PRAM ES eA RB SIR EA Ris HARARE AHRS A ELEN ASR EAR WEIR melee SRA LOTR REPENS ARTE DARE PIT AR AR AY 2 7 on BY oe EA BBR ZS a Te 52 FA A on 25 RNT PE SE A Bh I SS Sh This symbol indicates the product contains hazardous materials in excess of the limits established

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