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8.2 Optical unit

1. E Mark vent mar 1 Applies to Manual Run ONLY Run finished no 2 Does NOT apply to Manual Run KTAprime plus User Manual 11 0026 44 Edition AA 179 Reference information el Menu overview cont 5 Set Parameters Print out to Print Out Recorder Computer Method Loggdata Set Rec Out 1 UV Lamp pH Cond B Tmp Pr on Set Rec Out 2 B pH Cond B Tmn Pr Set Drop sync Active Set Rec Out 3 Cond yes pH Cond B Tmo Pr Memory Print Out Autoscaling no Press OK to start Printing Setup Analogue Out Printing Please wait Setup and Calibration Set Rec Out 1 UV Set UV Zero Level pH Cond B Tmn Pr monin Set Rec Out 2 B Set UV Range pH Cond B Tmp Pr 0 0005AUFS Set Rec Out 3 Cond bH Cond B Tmp Pr Set Cond Zero Level Set UV Anal Out LO ONC Larned ESAn OZI Set Cond Full Scale Set Cond Analog Out 50 00mS cm Set
2. Setup UV Set Show UV on Set Show UV On Reference information Set up pH calibration Set up pH temperature compensation See section 9 20 5 Calibrating the pH electrode optional Set up show conductivity 1 From the main menu select menu Set Parameters and press OK 2 Select sub menu Setup and calibration and press OK 3 Select sub menu Setup Cond and press OK 4 Select sub menu Set Show Cond The current status for showing conductivity is shown If on is shown current conductivity is displayed in the running display If off is shown no conductivity is displayed in the running display Press OK to change the setting 5 Change the setting as desired and press OK Setting the conductivity cell constant Calibrating the conductivity Setting the conductivity temperature compensation Setting the conductivity reference temperature See section 9 20 3 Calibrating conductivity Set up show UV Normally UV absorbance is shown in the running display If not required the UV absorbance display can be set to off 1 From the main menu select menu Set Parameters and press OK 2 Select sub menu Setup and calibration and press OK 3 Select sub menu Setup UV and press OK 4 Select sub menu Set Show UV The current status for showing UV is shown If on is shown current UV is displayed in the running display If off is shown no conductivity is displayed in the running display Press OK
3. Edit Breakpoint ml F mi 5 0 Base Set Concentration B a 20 0 B a ies Limit Set Flow Rate 2 0 ml min Folt Breakpoint Set Fraction Size 0 00 ml re ori Set Buffer Valve Pos Pos 1 a B on To a Set Inject Valve Pos LOAD End Method Set Peak Collect no Save Method i ZETO Edit time volume yes yes no E Mark Change Replace New time volume Edit time volume Run data displays 0 0 ml 5 0 ml 5 0 Save Breakpoint 2 RUN 7 8 min 22 6ml min 1 00MPa Delete Breakpoint 0 00002AU pH 8 50 0 00 ml 20 B 22 90mS cm Cond 78 8 Tc 22 4 C 2 Method Complete End run be 0 Om Press OK to complete no yes no Waste V waste BV 1 IV waste Set Concentration B B Set Gradient Set Flow Rate Print out Memory Print Out menus ves no Set Rec Out 1 UV UV pH Cond B Tmp Pr Set Rec Out 2 B UV pH Cond B Tmp Pr Set Rec Out 3 Cond UV pH Cond B Tmp Pr Autoscaling no Set Fraction Size Set Buffer Valve Pos os 1 Set Inject Valve Pos Press OK to start Set Peak Collect Printing 2 Autozero Printing Please wait
4. Injection valve and buffer valve e Check for external or internal leakage Replace channel plate and distribution plate when required Refer to section 9 13 Every 2 years Mixer e Replace the complete mixing chamber Refer to section 9 17 Superloop e Replace O rings Refer to section 4 in the instruction supplied with Superloop When required Monitor e Clean the conductivity flow cell according to section 9 11 e Clean the pH electrode flow cell according to section 9 18 Pump e Replace the check valve O rings according to section 9 16 e Clean the check valves If necessary replace the check valve O rings or the entire check valves according to section 9 16 9 2 Cleaning the system e Wipe the surface regularly with a damp cloth Do not allow spilt liquid to dry on the instrument e Remove dirt from the surface using a cloth and a mild cleaning agent e Let the system dry completely before using it AKTAprime plus User Manual 11 0026 44 Edition AA 121 9 Maintenance 122 9 3 Cleaning the system flow path WARNING When using hazardous chemicals make sure that the entire system has been flushed thoroughly with bacteriostatic solution e g NaOH and distilled water before service and maintenance Section 9 3 3 Monthly cleaning describes how to flush the system with a bacteriostatic solution Usually the column should be by passed with a
5. Measurement data to recorder On off signals to recorder To UV lamp high voltage KTAprime plus User Manual 11 0026 44 Edition AA 163 Reference information a 11 1 3 Fluid handling path The following flow diagram shows the positions of the components in AKTAprime plus fluid handling path Mixer G2 120 Pressure monitor System pump Male Male Flow Flow restrictor restrictor All capillaries are fitted at the factory The figure states the length in millimeters of the pre fabricated capillaries Tubing The table below lists the tubing fitted at delivery of AKTAprime plus Tubing Tubing Material Color Max Volume of Connected designation i d o d pressure 10cm Ai A B 3 2 mm 3 16 Teflon Clear 3 4 MPa 660 5 ul All inlet tubings from buffer vessels to switch valve AB 1 6 mm 1 8 Teflon Clear 2 MPa 201 1 ul From switch valve to pump W1 W3 1 0 mm 1 16 PEEK Brown 4 1 MPa 78 5 ul Waste tubing G1 G6 F1 0 75 mm 1 16 PEEK Green 2 6 MPa 44 2 wl From pump to fraction collector Union i d 1 16 PEEK Black 25 MPa Between conductivity flow cell male male orange and flow restrictor tubing Stop plug i d 1 16 PEEK Black Mixer inlet A 164 KTAprime plus User Manual 11 0026 44 Edition AA as Tubing connectors Reference information The table below lists the connectors and unions used Tubing designat
6. Vv Viewing the result E ths bc cas tel a leer a cue lente apes 29 WwW Wetted materials AKTAprime plus User Manual 11 0026 44 Edition AA Drop Design GSTPrep GSTrap HiLoad HiPrep HisPrep HisTrap HiTrap www ch romatog ra ph y PrimeView RESOURCE Sephacryl Superloop UNICORN KTA and KTAprime amershambiosciences com are trademarks of GE Healthcare Ltd GE tagline and GE monogram are trademarks of General Electric Company Microsoft and Windows are either trademarks or registered trademarks of GE Hea Ithca re Microsoft Corporation in the United States and or other countries i i All goods and services are sold subject to the terms and conditions of sale o Amersham Biosciences AB II goods and servi Id subj h d conditions of sale of fas the company within GE Healthcare which supplies them GE Healthcare Bj rkgata n 30 reserves the right subject to any regulatory and contractual approval if required to make changes in specifications and features shown herein or 751 84U ppsala discontinue the product described at any time without notice or obligation ontact your local ealthcare representative for the most curren d Contact your local GE Health tative for the most t Sweden information Purification and preparation of fusion proteins and affinity peptides comprising at least two adjacent histidine residues may require a license under US pat 5 284 933 and US pat 5 310 663 including corresponding foreign patents assign
7. KTAprime plus User Manual 11 0026 44 Edition AA 193 Reference information SSS Item Quant A C Code no pack Movable seal 1 A 19 7845 01 Protective jacket 50 ml 1 A 19 7849 01 Glass tube with thread and groove 1 A 19 7593 01 10 ml Glass tube with thread and groove 1 A 19 5165 01 50 ml Tubing kit for Superloop 10 ml 1 A 18 1113 83 Tubing kit for Superloop 50 ml 1 A 18 1113 84 Superloop 150 ml Superloop 150 ml complete 1 A 18 1023 85 Movable seal 1 A 18 1029 58 Inner end piece 1 A 18 1029 59 O ring inner end piece 2 C 18 1029 60 O ring movable seal 1 C 18 1134 49 Cables Mains cable 120 V 1 A 19 2447 01 Mains cable 240 V 1 A 19 2448 01 Mains distribution lead 0 3 m 1 A 18 1119 05 Signal cable KTAprime plus 1 A 18 1141 35 Connectors and unions Tubing connector 10 A 18 1112 49 Ti ji c inlet nut for o d 3 16 PEEK Ferrule for 3 16 o d tubing PEEK 10 A 18 1112 48 Tubing connector 10 A 18 1121 17 il iil g inlet nut for o d 1 8 PEEK pi Ferrule for 1 8 o d tubing PEEK 10 A 18 1121 18 Union 1 16 female M6 male PEEK 6 A 18 1112 57 Union luer female 1 16 male PEEK 2 A 18 1112 51 Union M6 female 1 16 male PEEK 8 A 18 1112 58 Union 5 16 female M6 male PEEK 3 A 18 1127 76 Union 1 16 male 1 16 male 10 A 18 1120 92 for 1 16 o d tubing PEEK Union 1 16 female 1 16 female 1 A 18 3855 01 for 1 16 o d tubing titaniu
8. Set Press Analogue Out 2 00 MPa 2 00 116 Setting the Cond analogue output 1 6 Select the Set Cond Analogue Out menu in the Setup Analogue Out menu by using the up and down buttons Current analogue settings are displayed zero and full scale values Press OK to access the settings menu The current setting is displayed Press OK Set the desired zero level value The range is 0 000 999 9 mS cm Press OK to acknowledge Press the down button to access the next settings menu The current setting is displayed Press OK Set the desired full scale value The range is 0 000 999 9 mS cm Press OK to acknowledge Press Esc to return to the Set UV Analogue Out menu Setting the pH analogue output Note The pH values for zero level and full scale must differ by at least 1 pH unit Note The zero level and full scale values can be calibrated in any order 1 6 Select the Set pH Analogue Out menu in the Setup Analogue Out menu by using the up and down buttons Current analogue settings are displayed zero and full scale values Press OK to access the settings menu Press OK Set the desired zero level value The range is pH 0 50 14 30 Press OK to acknowledge Press the down button to access the next settings menu Press OK Set the desired full scale value The range is pH 0 50 14 30 Press OK to acknowledge Press Esc to return to the Set UV Analogue Out menu Setting the Press analogue output
9. 1 Select the Set Press Analogue Out menu in the Setup Analogue Out menu by using the up and down buttons The current analogue setting is displayed full scale value Press OK to access the settings menu Set the desired full scale value The range is 0 00 1 00 MPa Press OK to acknowledge AKTAprime plus User Manual 11 0026 44 Edition AA Installing and modifying components 8 8 6 6 Printing curves directly after a run Selected run data is usually printed directly during the run However if a re print of the result is desired this can be done as a post run print out This procedure is described below The system has three analog output channels for printing the run data curves after the run You can choose to print out UV absorbance pH conductivity theoretical gradient B temperature or pressure Memory Print Out When the run is completed or if it has been aborted the prompt Memory Print yes yes no Out is displayed 1 To print out the run data select yes Otherwise select no Set Rec Out 1 Uv 2 Atthe Set Rec Out 1 menu select the parameter to be printed on channel 1 UV pH Cond B Tmp Pr Press OK Set Rec Out 2 UV 3 At the Set Rec Out 2 menu select the parameter to be printed on channel 2 UV pH Cond B Tmp Pr Press OK Set Rec Out 3 Uv 4 Atthe Set Rec Out 3 menu select the parameter to be printed on channel 3 UV pH Cond B Tmp Pr Press OK Autoscaling 5
10. e Wash the pump for eluent exchange without disturbing the column LOAD pos 1 WASTE pos 3 To To To column 7 From column column um From on From syringe syringe Waste es 158 AKTAprime plus User Manual 11 0026 44 Edition AA Reference information The geometry of the valves ensures that the flow path is completely swept so that solvent or sample memory effect is virtually non existing The switching parts are made of PEEK which ensures both long mechanical and chemical lifetime Monitor This is a high precision on line monitor for handling measurement data from the UV optical unit the conductivity cell and the pH electrode optional In combination with the flow cells the monitor offers fixed wavelengths of 214 nm Zn lamp optional 254 and 280 nm Hg lamp fast response high accuracy and reproducibility and low dead volumes UV optical unit The UV optical unit houses the lamp Zn or Hg the wavelength filter and the UV flow cell There are two flow E cells available optical path length 2 mm or 5 mm E optional The type of flow cell used depends on the j j UV flow cells with 2 sample amount applied and the size of the column and 5 mm path lengths The light beam from the lamp is directed through a double conical or a straight flow through cuvette 6 ul or 2 pl illuminated volume The photodetector current is fed to the signal processing circuitry inside the system The reference sig
11. The following illustration shows the location of the components in the system lt Column holder lt M Fow diversion Fraction valve collector A Monitor and Sample loop controller O _ oA rime plus LCD display Column OO Push buttons oS SOO UV flow cell Pump e zs Flow un restrictor 2 Conductivity cell Pressure Mixer Injection Buffer Switch valve Flow sensor valve valve restrictor 1 Buffer valve This is a rotary valve which has 8 positions The valve has a 360 rotating channel plate As the plate is turned by the motor the central port on the front is connected to one of the peripheral ports 1 8 allowing a clear liquid path The valve switching is controlled by the system by reading the actual position of the channel plate Central port The buffer valve is used on the low pressure side in the flow path before the pump It it used for switching between Peripheral port sample and buffer solutions The switching parts are made of PEEK which ensures long mechanical and chemical lifetime 156 KTAprime plus User Manual 11 0026 44 Edition AA Reference information Switch valve Flow diversion valve This is a 3 port 2 way valve supplied with CoM COM 24 V DC It has one permanently open port D marked COM and two ports marked NC nc NO NC normally closed and NO normally open i A Xe Port COM may be used as an inlet or an nea AINE outlet port There are two switch
12. conductivity between 50 Hz and 50 kHz giving maximum linearity and true conductivity values The conductivity is automatically calculated by multiplying the measured conductance by the flow cell s cell constant The cell constant is pre calibrated on delivery but can be measured with a separate calibration procedure This procedure is described in section 9 20 3 Calibrating conductivity One of the electrodes has a small temperature sensor for measuring the temperature of ent gt the eluent in the flow cell Temperature variations influence the conductivity and in some applications when highly precise conductivity values are required it is possible to program a temperature compensation factor that recalculates the conductivity to a set reference temperature Temperature sensor gt i 160 AkTAprime plus User Manual 11 0026 44 Edition AA Reference information Flow restrictor FR 902 There are two flow restrictors in the system each generating a backpressure of 0 2 MPa One is connected to port 5 on the injection valve to prevent buffer drainage when the injection valve is in position WASTE The second flow restrictor is connected after the 4 conductivity cell It prevents air bubbles being formed after the column when the pressure created in the column is released Compare to a cork in a bottle of champagne This must be allowed for when setting the maximum pressure limit in the methods For example
13. 0 01 0 2 0 02 0 01 0 1 KTAprime plus User Manual 11 0026 44 Edition AA Set Parameters Setup and calibration Setup UV Set Lamp Run Time Hg 2000 h Set Lamp Run Time Hg 2000 h 000 Set Parameters Setup and calibration Setup Temp Set Show Temp on Set Show Temp on on off Reference information Set up lamp run time When the UV lamp is replaced reset the Lamp Run Time counter 1 From the main menu select menu Set Parameters and press OK Select sub menu Setup and calibration and press OK Select sub menu Setup UV and press OK Select sub menu Set Lamp Run Time press OK Set the Lamp Run Time counter to zero with the dial Press OK to acknowledge Set up show temperature The display of the temperature in the conductivity flow cell shown in the running display can be enabled or disabled 1 From the main menu select menu Set Parameters and press OK Select sub menu Setup and calibration and press OK Select sub menu Setup Temp and press OK Select sub menu Set Show Temp The current status for showing temperature is displayed If on is shown current temperature is displayed in the running display If off is shown no temperature is displayed in the running display Press OK to change the setting Change the setting as desired and press OK AkTAprime plus User Manual
14. 01 HiTrap Butyl FF 1 ml 5x1ml 17 1357 01 HiTrap Butyl FF 5 ml 5x5ml 17 5197 01 HiPrep 16 60 Phenyl FF high sub 1x 20 ml 17 5095 01 HiPrep 16 60 Phenyl FF low sub 1x20ml 17 5094 01 HiPrep 16 60 Octyl FF 1x20ml 17 5097 01 HiPrep 16 60 Butyl FF 1x20ml 17 5096 01 CC AkTAprime plus User Manual 11 0026 44 Edition AA 191 Reference information Connection part schematic drawing 192 11 5 2 Consumables and accessories Item Quant A C Code no pack AkTAprime plus incl REC 112 1 C 11 0013 12 KTAprime plus without REC 112 1 C 11 0013 13 KTAprime plus without REC 112 1 C 11 0013 14 Japanese version PrimeView software kit 1 A 18 1152 84 incl CD manual and serial cable Pump Inlet check valve short 1 1 C 18 1172 43 Inlet check valve long 2 1 C 18 1172 44 Outlet check valve short 3 1 18 1172 45 Outlet check valve long 4 1 C 18 1172 46 O ring kit P 960 incl C 18 1172 53 1 42 x 1 52 mm connection part 7 4 x 1 2 mm check valves 12 Optical unit Hg lamp amp housing complete 1 C 18 1128 22 Zn lamp amp housing complete 1 C 18 1128 23 UV flow cell 5 mm 1 C 18 1128 24 UV flow cell 2 mm 1 C 18 1128 25 Filter 214 nm 1 C 18 0622 01 Filter 254 nm 1 C 18 0620 01 Filter 280 nm 1 C 18 0621 01 Filter 313 nm 1 C 18 0623 01 Filter 365 nm 1 C 18 0624 01 Filter 405 nm 1 C 18 0625 01 Filter 436 nm 1 C 18 0626 01 Filter 546 nm 1 C 18 0627 01 Filter wheel complete 1 A 18 0647 01 Conductivi
15. 1 Check that the conductivity flow cell cable is connected properly to the rear of the system 2 Check that the pump operates properly 3 If temperature compensation is being used check that the temperature sensor is calibrated and that the correct compensation factor is used 4 Check that the column is equilibrated If necessary clean the column 5 Check the operation of the mixer Baseline drift or noisy signal 1 There may be air in the flow cell Use a flow restrictor after the flow cell and check that the flow restrictor gives a back pressure of 0 2 0 05 MPa 2 Check for leaking tubing connections 3 Check that the column is equilibrated If necessary clean the column 4 Check the operation of the mixer and the pump Clean the flow cell according to the procedures in sections 9 11 Conductivity measurement with the same buffer appears to change over time 1 Clean the flow cell according to the procedures in sections 9 11 2 The ambient temperature may have changed Use a temperature compensation factor Waves on the gradient 1 Check that the pump and the valves are operating properly and are programmed correctly 2 Change to a larger mixing volume if necessary 3 Check the operation of the mixer Absolute conductivity value is wrong 1 Turn the flow cell so the end with screws faces the pH flow cell 2 Recalibrate the conductivity cell 3 Calibration solution 1 00 M NaCl not corr
16. 7 Ifthe UV curve is selected the current setting for auto scaling of the UV curve is shown Press OK to access the setting menu Autoscale UV no 8 Select the desired setting and press OK yes no The three selected curves are now printed Setting analog outputs See section 8 6 5 Setting analog outputs Calibrating the flow rate See section 9 20 1 Calibrating the pump Calibrating the pressure offset See section 9 20 2 Calibrating the pressure sensor Set up show pH Normally the pH is displayed in the running window If not required the pH display can be set to off Set Parameters 1 From the main menu select sub menu Set Parameters and press OK Setup and 2 Select sub menu Setup and calibration Press OK calibration Setup pH 3 Select sub menu Setup pH and press OK Set Show pH 4 Select sub menu Set Show pH The current setting for showing pH is on displayed If on is shown current pH is displayed in the running display If off is shown no pH is displayed in the running display Press OK to change the setting Set Show pH 5 Select the desired setting and press OK off on off SSS 168 KTAprime plus User Manual 11 0026 44 Edition AA Set Parameters Setup and calibration Setup Cond Set Show Cond on Set Show Cond on on off Set Parameters Setup and calibration
17. 8 2 5 Changing the lamp assembly optional section below SSS 100 AkTAprime plus User Manual 11 0026 44 Edition AA Installing and modifying components 8 n 8 2 3 Connecting the optical unit to the system If the optical unit has been disconnected from the system connect it as follows 1 Place the optical unit in the holder a 2 Secure it by tightening the screw in the holder 3 Connect the lamp cable to the socket UV I Lamp on the rear panel of the module Holder Optical unit 4 Connect the signal cable to the socket UV on the rear panel of the module 8 2 4 Connection to the column 1 Fix the optical unit directly under the column if possible Note Always position the optical unit with the filter wheel cover facing upwards 2 Connect the column outlet tubing directly onto the top of the optical unit using a fingertight connector and screw to finger tightness 3 Connect the optical unit outlet tubing onto the opposite hole in the flow cell Use fingertight connectors If no outlet tubing exists cut a piece of PEEK tubing i d 0 75 mm o d 1 16 The length should be 170 mm 4 Connect the other end of the tubing to the conductivity flow cell SSS AkTAprime plus User Manual 11 0026 44 Edition AA 101 ED Installing and modifying components 8 2 5 Changing the lamp assembly optional WARNING The system uses high intensity ultra violet light Do not remove the UV lamp while the system is running Before replacing a
18. If auto scaling of the UV curve is required select yes no yes no Otherwise select no 6 Press the set key on the recorder to define the start position for the print out 7 Press OK to print the curves AkTAprime plus User Manual 11 0026 44 Edition AA 117 8 Installing and modifying components Set Parameters Memory Print Out Print out to Recorder Computer 118 8 6 7 Printing curves before the next run The system stores the run data from the latest run Therefore post run print out can be done either at the end of the run as described in the precious section or before the next run using the Set Parameters menu as described below 1 2 10 Set the rec key on the recorder to off Press the home key to get the paper back to the start position Increase the chart speed value about ten times This is due to that the curves are printed ten times faster at the post run print out than during the actual run Set the rec key to on to enable the print out Set the pen key to position down Select main menu Set Parameters and press OK Select menu Memory Print Out and press OK At the Print out to menu select Recorder and press OK Select the desired parameters to be printed as in steps 2 5 on page 117 The print out will now overlay the previous curves Repeat the procedure above to select other parameters and print out the curves More information about the recorder is f
19. Re program the method 83 WARNING temp_cal will be changed 84 WARNING cond_cal will be changed Press OK to accept change 2 Press ESC to skip the change AKTAprime plus User Manual 11 0026 44 Edition AA 151 Troubleshooting Messages Action 85 WARNING conscale 1 The difference between 0 and 100 must be at least 0 1 mS cm 0 100 lt 0 1mS 2 Increase the span between zero and full scale setting See section 11 2 1 86 WARNING cond cell 1 Check that the conductivity cell is connected bad not connected 2 Recalibrate temperature 3 Ifthe problem remains replace the conductivity cell 87 WARNING pH probe 1 Check the pH electrode connection bad not connected 2 Clean the pH electrode 3 If the problem remains change the pH electrode Ba Eiscmical 1 Factory calibration for pH electrode is lost The monitor can still be used but ectrica halal may not meet the specifications for pH measurements Call for service 2 Call service 1 Factory calibration for conductivity is lost The monitor can still be used but 89 Electrical error hid Chat as may not meet the specifications for conductivity measurements Call for service 2 Call service 90 ATTENTION 1 Only visible to service personnel set lt 0mV first 91 WARNING bad pH ad value 92 WARNING electrode 1 Electro
20. Temp calibr will be changed Set Adjust Temp 25 0 C 25 0 AKTAprime plus User Manual 11 0026 44 Edition AA Maintenance ex Set up conductivity reference temperature 1 From the main menu select menu Set Parameters and press OK Select sub menu Setup and calibration and press OK Select sub menu Setup Cond and press OK Select sub menu Set Cond Ref Temp and press OK The current reference temperature value is shown 25 C is the default setting The range is 0 00 99 9 2C The current reference temperature value is displayed as default Adjust the reference temperature value setting as necessary and press OK 9 20 4 Calibrating the temperature sensor Calibration of the temperature sensor in the conductivity flow cell is only necessary if the monitor is used in high accuracy measurement or if the conductivity flow cell is replaced 1 Place the flow cell together with a precision thermometer inside a box or empty beaker to ensure that they are not exposed to draught Leave them for 15 minutes to let the temperature stabilize Read the temperature on the thermometer From the main menu select menu Set Parameters and press OK Select sub menu Setup and calibration and press OK Select sub menu Setup Temp and press OK Selec OK sub menu Set Adjust Temp The current temperature is shown Press A warning message is shown until confirmed by pressing OK The current adjustment value
21. UV lamp ensure that the lamp cable is disconnected from the rear of the system to prevent injury to the eyes If the mercury lamp is broken make sure thot all mercury is removed and disposed according to national and local environmental regulations Lamp housing end plate 1 Dot on lamp housing Symbols on filter housing Symbol on 6 filter wheel below lid 7 A 102 Use a screwdriver to detach the end plate by removing one and loosening the other of the two holding screws on the lamp housing to be removed Slide the lamp housing off the filter housing Detach the end plate as in step 1 above from the lamp housing to be fitted to the optical unit Slide the lamp housing onto the filter housing The lamp and signal cables should be on the same side As you slide the lamp housing into position depress the two pressure pads on the filter housing in sequence to facilitate the installation Refit the lamp housing end plate Slide the lamp housing firmly into place There will be a faint click when the housing is positioned correctly The Hg lamp housing can take up two positions one for 280 nm marked by O on the filter housing and the other marked by for all other wavelengths The Zn lamp housing has only one position Set the wavelength to be used by selecting lamp position indicated by a dot on the lamp housing in combination with the appropriate filter i e the dot on the lamp housing should be
22. a new cell Must be done when changing the flow cell constant PH electrode optional Every day The calibrations are made in the Setup and calibrations menu Set Parameters Setup and calibration 9 20 1 Calibrating the pump Calibrating the flow rate Calibrate the pump after replacing spare parts 1 Make sure no bubbles are trapped in the flow path 2 Immerse the inlet tubing A1 in a vessel filled with degassed buffer 3 Place the waste tubing from port 5 on the injection valve in an empty vessel the flow will automatically be diverted to port 5 during the calibration Start Pump Calibr 4 Select Start Pump Calibration in Setup and calibration menu Press OK 800 pulses Set Flow Rate 5 Enter the intended flow rate as calibration flow rate and press OK 10 ml min 2 0 6 Measure the volume of the water collected in the vessel Enter Collected 7 Enter the measured volume in ml in the Enter Collected Volume menu and Volume ml 2 05 press OK 8 Atthe Pump Calibrated OK menu press OK AkTAprime plus User Manual 11 0026 44 Edition AA 135 9 Maintenance Change Press Offset 1005 mV Set zero pressure to calib Press OK Set Parameters Setup and calibration Setup Cond Set Adj Cell Const 83 56cm Warning This will change cell calibr Set Adj Cell Const 83 56cm 83 55 Eee 136 9 2
23. adjacent to the symbol on the filter housing corresponding to the symbol on the filter wheel for the filter to be used A click will indicate that the filter is in position AKTAprime plus User Manual 11 0026 44 Edition AA Installing and modifying components 8 8 2 6 Changing the filter optional The Hg optics with 254 and 280 nm filters and the Zn optics with the 214 nm filter are delivered with filters installed If other filters are to be used install them as follows 1 If the Zn lamp is attached remove the lamp housing as described in section 8 2 5 Changing the lamp assembly optional 2 Remove the four screws in the filter housing Separate the filter housing from the detector housing Carefully remove the filter wheel from the filter housing If necessary remove the filter s from the filter wheel by pressing it them out e g with a small screwdriver Note Filters are sensitive optical components Never touch the optical surfaces or expose them to temperatures above 60 C Clean them with dry lens cleaning tissue and store them when not in use in the box in which they were supplied Heavy contamination may be removed by using a lens tissue dipped in ethanol Triangular aperture 5 Insert the filter s of choice into the filter wheel maximum 3 filters with the correct orientation the mirror side facing upwards and position over one of the three triangular apertures The filters snap in by pressing
24. and 1 150 ml samples respectively Sealing Internal electrolyte bridge containing 4 M KCI saturated with AgCI Silicone sealing Ag AgCl reference Pa electrode Internal annular coaxial ceramic reference junction External electrolyte bridge containing 1MKNO 3 O ring Glass electrode containing diluted buffered KCI O ring External annular coaxial ceramic reference junction AKTAprime plus User Manual 11 0026 44 Edition AA Reference information 11 2 Menus This section describes the Set parameters and Check menus It also contains an complete overview of the menus in AKTAprime plus 11 2 1 Set Parameters menus Turning the lamp on off We recommend the lamp be turned off to conserve lamp operating time when no measurement is being made A warm up time of 60 minutes is required to achieve full specifications However in most cases a warm up time of 15 minutes is sufficient 1 Select main menu Set Parameters and press OK Lamp 2 Select sub menu Lamp and press OK to access the setting menu on Lamp 3 Switch the lamp on off with the up and down buttons and then press OK on on off Setting drop synchronization If drop synchronization is active tube changes will only occur directly after a drop is registered by the drop counter to minimize spillage between tubes Drop
25. correct the fault call GE Healthcare Messages Action 1 Perform a new start up The preceding message may tell more about the 34 Start up failed cause Retry Call service 2 If not call service 1 Wrong value for averaging time set See section 11 2 1 35 WARNING wrong averaging time set 50 Electrical error 1 Call for service Call for service 57 Electrical error Call for service 75 Electrical error Call for service 1 Check that the fraction collector is not stuck 60 Tube switch a always active 2 Check the tube indicator 3 If the problem remains call service 61 No more tube is 1 Put more tubes in the fraction collector available 1 Check that a tube in the fraction collector touches the tube indicator 62 Check that the tube position is OK 2 Check the cable to the tube indicator 3 If the problem remains call service SRERROR NEA 1 Check the cable to the tube indicator 4 o drops Pa cad check sensor 7 2 Check that it is dripping If it flows continuously reduce the flow or turn off he Drop sync function 3 If the problem remains call service 65 ERROR 1 Restart the system Pump failure 2 If the problem remains call service 66 Too short time between feeds Reduce the flow or increase the fraction size 67 ERROR Injection valve failure Call service 68 ERROR Buffer valve failure Call service 150 AKTAprime plus User Manual 11 0026 44 Edition AA Troub
26. flow cell is recalibrated after cleaning Note The conductivity temperature compensation must not be used when adjusting the cell constant Set the Set Cond Temp Comp to 0 see page 138 The temperature sensor must be calibrated before adjusting the cell constant see page 139 1 Prepare a calibration solution of 1 00 M NaCl 58 44 g l Let the solution stand until it is at room temperature This is important for exact measurements 2 Fill the flow cell completely with the calibration solution by pumping at least 15 ml through the cell with a syringe 3 Stop the flow and wait 15 minutes until the temperature is constant in the range 20 30 C 4 Read the conductivity value displayed and compare it with the theoretical value from the graph below at the temperature of the calibration solution If the displayed value and the theoretical value correspond no further action is required If the values differ proceed with the actions below SSS AkTAprime plus User Manual 11 0026 44 Edition AA 137 9 Maintenance Set Parameters Setup and calibration Setup Cond Set Adjust Cond 80 32mS cm Warning This will change cell calibr Set Adjust Cond 80 32mS cm 83 55 Set Parameters Setup and calibration Setup Cond Set Cond Temp Comp 0 0 Set Cond Temp Comp 0 0 0 0 138 10 From the main menu Conductivity of
27. in 0 1 M NaOH and again in 0 1 M HCI Each immersion is for a period of 5 minutes Rinse the electrode tip in distilled water e Oil or grease films Wash the electrode tip in liquid detergent and water If the film is known to be soluble in a particular organic solvent wash with this solvent Rinse the electrode tip in distilled water e Protein deposits Dissolve the deposit by immersing the electrode in a 1 pepsin solution in 0 1 M HCI for five minutes followed by thorough rinsing with distilled water It these procedures fail to rejuvenate the electrode the problem is most likely a clogged liquid junction Use the following procedure 1 Heata 1M KNO solution to 60 80 C 2 Place the electrode tip in the heated KNO solution 3 Allow the electrode to cool while immersed in the KNO solution before re testing If these steps fail to improve the electrode response replace the electrode 9 19 Replacing the pH electrode optional See section 8 5 pH flow cell and electrode optional AKTAprime plus User Manual 11 0026 44 Edition AA Maintenance ex 9 20 Calibrations Follow the calibration schedule below to maintain accurate measurements Component How often Pump Only necessary after replacing spare parts Pressure offset When required Conductivity flow cell Cell constant Only necessary if specific conductivity with high accuracy is measured Temperature Must be done when changing the flow cell Entering
28. limit The gel packing pressure limit is always unaffected by a flow restrictor The figure below shows an example of how a flow restrictor affects the packed column at a hypothetical flow rate of 1 ml min The flow rate generates a pump pressure reading of 0 3 MPa With no restrictor mounted this pressure equals the pressure drop over both the gel and the column hardware AkTAprime plus User Manual 11 0026 44 Edition AA 161 Reference information SEs j Flow rate 1 ml min Pump pressure fay Pump pressure reading 0 3MPa _ Bs reading 0 5 MPa Gel top pressure 0 3 MPa IL L 05 MPa LL Gel top pressure Column hardware Column hardware Gel pressure drop pressure 0 3 MPa pressure 0 5 MPa Gel pressure drop 0 3 MPa 0 3 0 0 0 3 0 2 0 3 MPa Gel bottom pressure J mme mm Ol Gel bottom pressure 0 0 MPa 0 2 MPa Flow restrictor When a flow restrictor generating a backpressure of 0 2 MPa is mounted after the column and the same flow rate is used the pressure over the column hardware is affected and will be 0 5 MPa Hence the pump pressure gauge reading will be 0 5 MPa However the pressure drop over the packed gel bed is still 0 3 MPa Fraction collector The fraction collector can be used for both small scale and preparative scale purifications It collects up to 175 fractions in 12 mm diameter tubes and up to 95 fractions in 10 18 mm diameter tubes The fraction coll
29. new flow cell the system must be calibrated with the new cell constant written on the flow cell package See section 9 20 3 Calibrating conductivity If the cell constant is not known it can be determined AkTAprime plus User Manual 11 0026 44 Edition AA 127 9 Maintenance 128 9 13 Replacing plates in the rotary valves A replacement kit for each valve is available Refer to 11 5 Ordering information Channel plate Distribution plate Injection valve Buffer valve 1 Make sure that the valve is in position 1 see figure and then disconnect all tubings 2 Remove the four screws on the front using a 3 mm hex wrench Loosen each one equally in turn so that the distribution plate comes off parallel to the valve body 3 Slide the screws out 4 Remove the distribution plate containing the ports 5 Remove the old channel plate and insert the new one 6 Remounta new distribution plate so that the text 3 injection valve or i o buffer valve is horizontal and to the right of the central tubing connection Using a hex wrench tighten the four screws in turn a little at a time until the distribution plate is fixed to the valve body AKTAprime plus User Manual 11 0026 44 Edition AA Maintenance 9 as 9 14 Removing and assembling the pump The connection part should be removed to allow access to the check valves and the O rings 9 14 1 Removing the connection part Bef
30. pressure is assumed Note Chemical influences are time and pressure dependent Unless otherwise stated all concentrations are 100 Chemical Exposure Comments Acetic acid 0 1 M OK Acetone 1 OK Aqueous buffers pH 2 12 OK Decon 90 10 OK For washing only Ethanol 20 OK Ethanol 96 OK For washing only Ethylene glycol OK Formic acid 1 OK Guanidin 6 M OK HCI 0 1 M OK Isopropanol 30 OK Lysozyme 2 mg ml OK Methanol 20 OK NaOH 0 1 M OK NaOH 1 M OK For washing only SDS 10 OK Short term use TFA 0 2 OK Triton X 2 OK Short term use Urea 8 M OK SSS AkTAprime plus User Manual 11 0026 44 Edition AA 187 Reference information 11 5 Ordering information 11 5 1 Recommended columns lon Exchange Columns Column name Pack size Code no HiTrap Q HP 1 ml 5x1lm 17 1153 01 HiTrap Q HP 5 ml 5x5m 17 1154 01 HiTrap SP HP 1 ml 5x1lm 17 1151 01 HiTrap SP HP 5 ml 5x5m 17 1152 01 HiTrap IEX Selection kit 7x1m 17 6002 33 HiTrap Q FF 1 ml 5x1lm 17 5053 01 HiTrap Q FF 5 ml 5x5m 17 5156 01 HiTrap SP FF 1 ml 5x1lm 17 5054 01 HiTrap SP FF 5 ml 5x5m 17 5157 01 HiTrap DEAE FF 1 ml 5x1lm 17 5055 01 HiTrap DEAE FF 5 ml 5x5m 17 5154 01 HiTrap CM FF 1 ml 5x1lm 17 5056 01 HiTrap CM FF 5 ml 5x5m 17 5155 01 HiTrap ANX FF high sub 1 ml 5x1m 17 5162 01 HiTrap ANX FF high sub 5 ml 5x5m 17 5163 01 HiTrap Q XL 1 ml 5x1lm 17 5158 01 HiTrap Q XL 5 ml 5x5m 17 5159 01 HiTrap SP XL 1 ml 5x1lm 17 5160 01 HiTr
31. sensor The pressure in the system is continuously measured by the pressure sensor located next to the pump The pressure is shown on the display during the run A maximum pressure limit can be set to protect the column Mixer This is a dynamic single chamber mixer with interchangeable mixer chambers The system is delivered with a 2 ml chamber The eluents are mixed in two steps t 1 Premixing in a static mixer with a small volume 22 ul 2 2 Dynamic mixing in a chamber with a rotating stirrer A mixer motor inside the system spins a magnet at 600 rpm which 1 causes the stirrer in the mixing chamber to rotate x For optimal gradients at high flow rates a larger mixer chamber is required Other mixer chambers with 0 6 5 and 12 ml mixer volumes are available as accessories When using eluents that are more difficult to mix such as isopropanol and water or if the pH and conductivity readings indicate uneven mixing of your buffers unstable readings a larger mixer volume will give better mixing Injection valve A 7 port motorized rotary valve is used as sample injection valve It has a valve body with a rotating central core the channel plate As the channel plate is rotated by the motor different ports are connected The valve has three different operating positions which make it possible to e Load a sample loop without disturbing column equilibration e Wash the sample loop while the column is in operation
32. setting for channel 1 is displayed first Press the up and down buttons to display the settings for channels 2 and 3 2 Select the desired channel and press OK In this example channel 1 is selected 3 Select the desired parameter and press OK Note The analogue output level for Tmp has a fixed set value 0 C corresponds to 0 Vand 50 C corresponds to 1 0 V Setting the UV analogue output 1 Select the Set UV Analogue Out menu in the Setup Analogue Out menu by using the up and down buttons Current analogue settings are displayed zero and full range values Allowed full range values are 0 0001 0 0002 0 0005 0 001 0 002 0 005 0 01 0 02 0 05 0 1 0 2 0 5 1 0 2 0 and 5 0 Zero level is set as a percentage of full scale 2 Press OK to access the settings menu The current setting is displayed Press OK 3 Set the desired zero level value Press OK to acknowledge 4 Press the down button to access the settings menu The current setting is displayed Press OK 5 Set the desired full range value Press OK to acknowledge 115 8 Installing and modifying components Set Cond Analogue Out 00 00 50 00mS cm Set Cond Zero Level 0 00mS cm 0 00 Set Cond Full Scale 50 00mS cm 50 00 Set pH Analogue Out pH 0 00 14 00 Set pH Zero Level pH 0 00 00 00 Set pH Full Scale pH 14 00 14 00 Set Press Analogue Out 1 00 MPa 1 00
33. synchronization operates in all fraction collection modes time volume and drop Drop synchronization is only possible at flow rates up to 3 ml min If the flow limit is exceeded an error message is displayed 1 Select main menu Set Parameters and press OK Set Drop Sync Active 2 Select sub menu Set Drop Sync Active The current setting is shown The yes setting will apply to all subsequent manual and method controlled operation until a new value is set Press OK to change the setting Set Drop Sync Active 3 Select the desired setting and press OK yes yes no Memory print out Measurement data from the last run can be printed to a recorder or a computer Three channels are available for printing the curves that correspond to UV absorbance pH conductivity concentration of the B buffer temperature and pressure 1 Select main menu Set Parameters and press OK Memory Print Out 2 Select sub menu Memory Print Out and press OK AkTAprime plus User Manual 11 0026 44 Edition AA 167 Reference information es Print out to 3 Select printing to a recorder or a computer and press OK Recorder Computer Set Rec Out 1 Uv 4 The current setting for channel 1 is displayed Press OK to access the setting menu Set Rec Out 1 Uv 5 Select the parameter to be printed on channel 1 and press OK UV pH Cond B Tmp Pr 6 Repeat steps 4 and 5 for channels 2 and 3 Autoscale UV no
34. that are approved or supplied by GE Healthcare may be used for maintaining or servicing the system WARNING Use ONLY tubings supplied by GE Healthcare to ensure that the pressure specifications of the tubings are fulfilled DDD D gt DDD AkTAprime plus User Manual 11 0026 44 Edition AA 119 9 Maintenance WARNING If the system is turned the external capillaries and other tubing may become entangled in nearby objects and be pulled from their connections causing leakage 9 1 1 Preventive maintenance schedule Interval Action Daily System Inspect the complete system for eluent leakage The system can be left filled with buffer overnight If you are not using the separation unit for a few days 1 Wash the flow path with distilled water 2 Remove the column and the pH electrode optional 3 Wash the flow path with 20 ethanol and store it in 20 ethanol Make sure that all tubing and all flow paths used are rinsed Pump Check for leakage If there are signs of liquid leaking from the pump Check the tubing connections Check the O rings in the connection part Replace the O rings if necessary If there are signs of erratic or pressure pulsation flush the pump with 100 methanol and then with distilled water pH electrode Calibrate the pH electrode according to the section Calibrating the pH electrode optional Every week Inlet filters Check
35. them quite firmly Do not touch the filter surface 6 Remove the circular plastic band showing the wavelengths 7 Remove labels from the band if necessary 8 Place the correct labels on the band with the label designation facing outwards Ensure that the label position corresponds to the filter position i e the label should be placed opposite the filter 9 Reassemble the circular plastic band with the filter wheel peg fitting into the band notch 10 Check that all filters are clean Insert the filter wheel back into the filter housing Note The filter wheel can be placed only in one correct position 11 Reassemble the filter housing with the detector housing by fastening the four screws AkTAprime plus User Manual 11 0026 44 Edition AA 103 ED Installing and modifying components 104 8 3 Conductivity cell To install a conductivity cell Conductivity Holder cell Union Flow Lock screw restrictor 1 Connect the conductivity cell to the flow restrictor using the union 2 Insert the cell into the holder and secure it with the lock screw Note When the conductivity flow cell is used with the pH electrode place the conductivity flow cell and select its flow direction so that the screw head end of the flow cell faces the flow restrictor 3 Connect the conductivity cell to the socket Conductivity Flow Cell on the rear panel of the system 4 Connect the tubing with fingertight connec
36. when using a column with a pressure limit of 0 3 MPa the pressure limit in the method should be set to 0 5 MPa This is described below in detail Two types of pressure limits All chromatography columns have two types of pressure limits one for the column hardware and one limit for the gel packing The pressure affecting the column hardware depends on the backpressure generated after the column by for example flow cells flow restrictors and tubing When the pressure limit for the column hardware is exceeded the column will start leaking Empty chromatography columns from GE Healthcare are all supplied with data on the column hardware pressure limit The pressure affecting the gel packing is dependent on the flow rate and viscosity of the buffer When the flow rate is too high and or a high viscosity buffer is used the pressure limit for the packed gel bed can be exceeded the gel pressure limit is the maximum allowable pressure drop over the packed gel bed When the pressure limit is exceeded the gel particles are forced to the bottom of the column and cause the backpressure to increase This leads to a collapse of the packed bed and the risk of poor chromatographic performance The pressure limit data referring to the gel packing is found in the column instructions delivered with all prepacked columns from GE Healthcare How the flow restrictor affects the pressure limits The flow restrictor only affects the column hardware pressure
37. 0 2 Calibrating the pressure sensor Calibrating the pressure offset The zero pressure should be calibrated when required 1 4 Make sure that the pressure sensor is exposed to atmospheric pressure only i e no backpressure Select the Change Press Offset menu in the Setup and calibration menu Press OK At the Set zero pressure to calib menu press OK The calibration only takes a few seconds Press OK at the Calibrating Offset Done menu 9 20 3 Calibrating conductivity Set up adjust cell constant After replacing the flow cell the cell constant has to be set The cell constant is written on the cell packaging 1 From the main menu select menu Set Parameters and press OK Select sub menu Setup and calibration and press OK Select sub menu Setup Cond and press OK Select sub menu Set Adj Cell Const The current cell constant is shown Press OK A warning message is shown until confirmed by pressing OK The current cell constant is displayed as default Enter the new cell constant as read from the packaging and press OK The range is 0 1 300 0 cmt AKTAprime plus User Manual 11 0026 44 Edition AA Maintenance ex Set up adjust conductivity Normally it is not necessary to adjust the cell constant because the flow cell is pre calibrated on delivery Adjustment is only necessary when replacing the conductivity flow cell with a flow cell whose cell constant is unknown We recommend that the conductivity
38. 0 Start RUN 1 Does not apply to To RUN data displays Gel filtration Buffer exchange and Print out menus see page 179 AKTAprime plus User Manual 11 0026 44 Edition AA Reference information Say Menu overview cont 2 Run Stored Method Run Stored Method Number 14 Run Stored Method Run Stored Method 1 From System PC 1 From PC No 78 Run Stored Method 1 From System No 14 Press OK to start run To RUN data displays and Print out menus Copy Method see page 179 Copy Method From Copy Method From 1 System PC 1 PC No 76 Copy Method From System No 16 Copy Method To Copy Method To 7 System PC 1 PC Free No 76 Copy Method To System Free No 16 Method occupied Copy Method Press OK Copy Method Clear it yes no 1 PC 76 gt System 17 Please wait Copy Method Press OK 1 System 17 gt PC 76 Copy Method Press OK 1 PC 53 gt PC 87 Copy Method Press OK 1 These displays appear only if a System 17 gt System 28 computer running PrimeView is connected to the system KTAprime plus User Manual 11 0026 44 Edition AA 177 Reference information Ol Menu overview cont 3 Set Method Base ml Set Concentration B
39. 0 V autorange 50 60 Hz Power consumption Max 90 VA Fuse specification T 1 0 AH 250 VAC approved type not replaceable by operator Dimensions Hx Wx D 530 x 400 x 450 mm Weight 13 kg Environment 4 to 40 C 10 95 relative humidity non condensing 84 106 kPa 840 1060 mbar atmospheric pressure Compliance with standards The declaration of conformity is valid for the instrument only if it is e used in laboratory locations e used inthe same state as it was delivered from GE Healthcare except for alterations described in the User Manual e connected to other CE labelled GE Healthcare modules or other products as recommended Safety standards This product meets the requirement of the Low Voltage Directive LVD 73 23 EEC through the following harmonized standards e EN 61010 1 e IEC 61010 1 e CAN CSA C22 2 No 61010 1 e UL61010 1 AKTAprime plus User Manual 11 0026 44 Edition AA Reference information EMC standards This device meets the requirements of the EMC Directive 89 336 EEC through the following harmonized standards e EN 61326 emission and immunity e N 55011 GR 2 Class A emission e This device complies with part 15 of the FCC rules emission Operation is subject to the following two conditions 1 This device may not cause harmful interference 2 This device must accept any interference received including interference that m
40. 1 00 M NaCl at 20 30 C select menu Set 97 Parameters and press OK Select sub menu Setup and calibration and press OK Select sub menu Setup Cond and press OK 90 Select sub menu Set Adjust Cond The current conductivity value is shown Press OK A warning message is shown until confirmed by pressing OK 85 Conductivity mS cm The current value is displayed as default Enter the theoretical conductivity value according to the graph and press OK The new cell constant is automatically calculated The range is 20 25 30 1 000 999 9 mS cm Temperature tei 80 Set up conductivity temperature compensation 1 From the main menu select menu Set Parameters and press OK Select sub menu Setup and calibration and press OK Select sub menu Setup Cond and press OK Select sub menu Set Cond Temp Comp and press OK The current temperature compensation factor is shown 0 0 means that the compensation is off default setting The range is 0 0 9 9 The current compensation factor is displayed as default Adjust the compensation factor setting as necessary and press OK KTAprime plus User Manual 11 0026 44 Edition AA Set Parameters Setup and calibration Setup Cond Set Cond Ref Temp 25 0 C Set Cond Ref Temp 25 0 C 25 0 Set Parameters Setup and calibration Setup Temp Set Adjust Temp 25 0 C Warning
41. 10 Fax 0165 580 401 e Norway Tel 815 65 555 Fax 815 65 666 e Portugal Tel 21 417 7035 Fax 21 417 3184 e Russia amp other C I S amp N I S Tel 7 095 232 0250 956 1137 Fax 7 095 230 6377 e South East Asia Tel 60 3 8024 2080 Fax 60 3 8024 2090 e Spain Tel 93 594 49 50 Fax 93 594 49 55 e Sweden Tel 018 612 1900 Fax 018 612 1910 Switzerland Tel 0848 8028 12 Fax 0848 8028 13 UK Tel 0800 616928 Fax 0800 616927 e USA Tel 800 526 3593 Fax 877 295 8102 imagination at work User Manual 11 0026 44 AA E Elanders Tofters 2005
42. 11 0026 44 Edition AA 171 Reference information Set up Delay UV to Frac The delay volume in ul is the volume of the flow path between the UV flow cell and the fraction collector If the set delay volume is correct the event marks on the UV absorbance curve will be synchronized with the tube changes Calculate the volume of the tubing from the UV flow cell to the very end of the tubing The volume of 0 75 mm i d tubing is 44 2 ul 10 cm The default delay volume is 380 ul If using a pH electrode 100 ul should be added to the delay volume 480 ul provided that the tubing lengths remain unchanged Set Parameters 1 From the main menu select menu Set Parameters and press OK Setup and 2 Select sub menu Setup and calibration and press OK calibration Set Delay UV to Frac 3 Select sub menu Set Delay UV to Frac The current delay volume is shown 380 ul Press OK to change the volume Set Delay UV to Frac 4 Change the setting as desired and press OK 380 jl 380 Set up mixer chamber volume If the mixer chamber is exchanged for a chamber with another volume the new chamber volume must be set On delivery the chamber volume is set to 2 0 ml If a larger chamber volume is used the wash and priming volumes must be increased accordingly Set Parameters 1 From the main menu select menu Set Parameters and press OK Setup and 2 Select sub menu Setup and calibrat
43. 12 is the ON OFF signal and pin 15 is signal ground 9 Connect the mains cable to a properly grounded mains socket WARNING The system must be connected to a grounded mains socket to prevent system parts from becoming live 10 Turnon the mains power to the recorder AkTAprime plus User Manual 11 0026 44 Edition AA 111 8 Installing and modifying components 8 6 2 Loading the chart paper roll To load the chart paper roll 1 Unpack the paper roll supplied with the recorder and put into place 1 Make sure the paper roll is positioned so that the printed grid faces the user during normal operation Lift the plastic transparent ruler 2 by pushing it gently upwards so that it rests over the top of the recorder Leave it in this position 112 Pull the paper forward It should be parallel to the housing Use your fingertips to fit the holes in the edge of the paper over the sprockets Do this on the right hand side first and then on the lefthand side 3 This is important because the holes in the right side guarantee exact paper positioning whereas the holes on the left side only support transport When the paper is correctly positioned over both sprocket wheels hold the paper in position and lower the ruler Use both hands to gently apply pressure to both the left and right edge of the ruler A double click indicates that the ruler is properly positioned AKTAprime plus User Manual 11 0026 44 Edition
44. 20 B Set Gradient Set Length off Set Flow Rate 3 0 ml min Set Fraction Base ml Set Fraction Size 00 0 ml Set Pressure Limit 1 00 MPa 0 00 ml Set Target 00 B 00 Set Buffer Valve Pos Pos 1 Set Inject Valve Pos Load Start Run 178 Check Communication Check Autozero AZ 0 0000AU Check Lamp Run time Hg00000h Zn00000h Check Lamp Intensity R 5 5 S 5 7 mV Check Pump Run time 123456h Check Pumped Volume 123456789 ml Check Tube Shifts 123456 Check Valve Shifts BV 123456 IV 123456 Check Recorder Check Service Mode Press OK to start run To RUN data displays and Print out menus see page 179 Telephone Service 012345678901234567 Contract Number 012345678901 Serial Number 01234567 YM 012345 AKTAprime VX XX XX Date of maintenance DD MMMM YYYY Buzzer Test AKTAprime plus User Manual 11 0026 44 Edition AA Program Method Free Methods 25 o edit edit clear Reference information Menu overview cont 4 Method Occupied Clear Method 09 yes yes no Sel Method Used 09 Set Method Base
45. 3 of setting pressure gt 0 1 MPa flow rate gt 0 4 ml min Gradient composition accuracy reproducibility 3 at 0 1 50 ml min 5 95 B 1 0 at 0 1 50 ml min 5 95 B Leakage lt 1 0 l min complete system Pressure sensor range scale error offset error 0 1 1 MPa 2 0 02 MPa UV measurement Wavelengths Hg lamp fixed by changing filter option 254 and 280 nm 313 365 405 436 and 546 nm Zn lamp optional 214nm Absorbance range 0 001 5 0 AU Autozero range 0 2 2 0 AU Baseline adjust Adjustable 0 100 of full scale Linearity lt 3 up to 2 AU at 254 nm lt 5 at other wavelengths except 280 nm Static noise short term 40x10 6 AU at 254 nm long term 40x10 AU at 254 nm Static drift 100x10 AU hour at 254 nm Flow sensitivity 2x10 AU min ml AKTAprime plus User Manual 11 0026 44 Edition AA 181 Reference information UV flow cell 2mm Flow rate 0 100 ml min Max pressure 4 0 MPa Max backpressure 0 05 MPa at 100 ml min Liquid temperature range 4 to 60 C Optical path length 2mm Cell volume 2 wl 30 ul detector volume UV flow cell 5 mm optional Flow rate 0 20 ml min Max pressure 4 0 MPa Max backpressure 0 02 MPa at 20 ml min Optical path length 5mm Cell volume 6 ul 10 ul detector volume Conductivity measurement Conductivity range 1
46. 86 01 HisTrap FF crude 5 ml 100 x 5 ml 17 5286 02 GSTrap HP 1 ml 5x1ml 17 5281 01 GSTrap HP 1 ml 100 x 1 mlt 17 5281 02 GSTrap HP 5 ml 1x5ml 17 5282 01 GSTrap HP 5 ml 5x5ml 17 5282 02 GSTrap HP 5 ml 100 x 5 mlt 17 5282 05 GSTrap FF 1 m 5x1ml 17 5130 01 GSTrap FF 1 m 2x1ml 17 5130 02 GSTrap FF 1 m 100 x 1 mlt 17 5130 05 GSTrap FF 5 m 1x5ml 17 5131 01 GSTrap FF 5 m 5x5ml 17 5131 02 GSTrap FF 5 m 100 x 5 mlt 17 5131 05 GSTPrep FF 16 10 1x20ml 17 5234 01 GSTrap FF crude 1 ml 5x1ml 11 0004 60 GSTrap FF crude 1 ml 100 x 1 ml 11 0004 61 GSTrap FF crude 5 ml 5x5ml 17 5284 02 a 190 AkTAprime plus User Manual 11 0026 44 Edition AA Reference information SS _ _ Column name Pack size Code no GSTrap FF crude 5 ml 100 x 5 mlt 17 5284 03 HiTrap Benzamidine FF high sub 1 ml 2ximl 17 5143 02 HiTrap Benzamidine FF high sub 1 ml 5x1ml 17 5143 01 HiTrap Benzamidine FF high sub 5 ml 1x5ml 17 5144 01 1 Pack size available on special order Hydrophobic Interaction Columns Column name Pack size Code no HiTrap HIC Selection kit 5x1ml 17 1349 01 HiTrap Phenyl FF high sub 1 ml 5x1ml 17 1355 01 HiTrap Phenyl FF high sub 5 ml 5x5ml 17 5193 01 HiTrap Phenyl FF low sub 1 ml 5x1ml 17 1353 01 HiTrap Phenyl FF low sub 5 ml 5x5ml 17 5194 01 HiTrap Phenyl HP 1 ml 5x1ml 17 1351 01 HiTrap Phenyl HP 5 ml 5x5ml 17 5195 01 HiTrap Octyl FF 1 ml 5x1ml 17 1359 01 HiTrap Octyl FF 5 ml 5x5ml 17 5196
47. AA Installing and modifying components 8 8 6 3 Installing the pens Recorder REC 112 has a writing system that is very simple and convenient It uses reliable fibre tip pens of the disposable cartridge type 1 which leave a very fine trace and do not bleed To install the pens 1 Remove the cap 2 Gently push the pen into the plastic pen holder 2 avoiding sidewards or upwards pressure Note To prevent the pens from drying when the recorder is not in use cover the tips of the pens using the caps provided 18 1004 23 ZO q W 18 1004 22 KTAprime plus User Manual 11 0026 44 Edition AA 113 ED Installing and modifying components E 8 6 4 Preparing the recorder for a run 8 p S x z 2 i 1 2 Alien 8 s J a 0 16 s ell mae 10 U When J gt a 5 fo me Baise 9 p ae AM 7 z i 13 15 14 11 12 3 2 6 4 5 1 Turn on the mains power to the recorder 2 Set the zero keys 4 to the down position 3 Select a suitable low chart speed 0 5 2 mm min with the chart speed selector 8 and the mm s mm min key 9 4 Set both pens 6 to position down Use the adjust knob 5 to make a coarse zero adjustment to the right hand zero on the chart 5 Set rec 7 to position on and make a final zero adjustment 6 Set rec to position off Use the forward feed key 11 12 to align the short nib pen with a grid line and p
48. ED Installing and modifying components E 8 2 Optical unit 8 2 1 Connecting the optical unit holder Hook the holder into the slot on the right hand side of the system Secure it by pushing up the slide clamp 8 2 2 Changing UV flow cell A preparative 2 mm flow cell is included in the system An analytical 5 mm flow cell is available as an accessory The flow cell can be changed when required for example from 2 mm to 5 mm to increase the sensitivity or from 5 mm to 2 mm to decrease the sensitivity Change the flow cell as follows 1 Disconnect the inlet and the outlet capillaries from the flow cell 2 Loosen the flow cell by turning the locking nut and remove it 3 Remove the protective cover from the Protective cover for old flow cell and transfer it to the new flow cell flow cell 4 Insert the new flow cell into the detector housing from above O ring 2MM Can Note The flow cell can only be placed Can in one correct position 5MM 5 Secure the flow cell by turning the locking nut until it reaches its stop position Note Ifthe locking nut is not tightened rae sufficiently the monitor will function poorly e g drifting base line 6 Place the protective cover around the flow cell to Locking protect the electronics inside the optical unit nut from liquid spillage Note Ensure that the Hg lamp position and the filter are selected according to the wavelength to be used This is described in the
49. KNO3 Do NOT store in water only If you are not using the system for a few days or longer 1 Wash all tubing and flow paths used with deionized water for example by running the System Wash Method with all tubing inlets in water 2 Replace the column with a bypass capillary 3 Replace the pH electrode optional with a dummy pH electrode 4 Wash the system with 20 ethanol and store it in 20 ethanol The UV flow cell can also be stored dry by flushing as above with distilled water and then 20 ethanol through the flow cell Replace the red protective caps Never use compressed air as this may contain droplets of oil pH electrode optional The pH electrode should always be stored in a 1 1 mixture of pH 4 buffer and 2 M KNO when not in use After removing the pH electrode from the flow cell insert a dummy electrode in the flow cell 9 3 3 Monthly cleaning WARNING NaOH is injurious to health Avoid spillage Clean the system every month before service and maintenance or when problems such as ghost peaks occur The system is cleaned as follows 1 Disconnect the column and replace it with a suitable capillary 2 Put all tubing inlets in 1 M NaOH 3 Run System Wash method for all inlet tubings 4 Flush the whole system with 1 M NaOH for 20 minutes 1 ml min 5 Immediately repeat steps 3 and 4 with distilled water to rinse the system of NaOH AkTAprime plus User Manual 11 0026 44 Edition AA 123 9
50. Maintenance 9 3 4 Other cleaning considerations After repeated separation cycles contaminating material may progressively build up in the system and on the columns This material may not be removed by the cleaning step described above The nature and degree of contamination depends on the sample and the chromatographic conditions employed 9 4 Moving the system CAUTION Never lift the system by the components mounted on the system chassis WARNING When using hazardous chemicals make sure that the entire system has been flushed thoroughly with bacteriostatic solution e g NaOH and distilled water before service and maintenance WARNING If the system is turned the external capillaries and other tubing may become entangled in nearby objects and be pulled from their connections causing leakage Check Lamp Intensity R 215 5 214 7mV 124 Before moving the system ensure that all cables and capillaries connected to peripheral equipment and liquid containers are disconnected Lift the system by placing your fingers in the gap between the base and the work bench surface grasping firmly and lifting 9 5 Checking the UV monitor 9 5 1 Checking lamp intensity 1 Select menu Check and press OK 2 Select menu Check Lamp Intensity If R lt 300mV for 254 nm R lt 150mvV for 280 nm or R lt 150mV for 214 nm replace the lamp assembly according to section 8 2 5 Changing the lamp assembly
51. Pressure sensor PrimeView starting Printing a simple report Programming line by line Programming using method templates Pump applying large samples with assembling removing trapped air troubleshooting Purging the pump and inlet tubing Purification work flow R Recommended COMMAS sereia su tes ensertecesbagstta eee e E aneceaey 188 Recorder REC 112 installation preparation Recorder signal cable Removing trapped air from the pump Restart after power failure Running the system manually S Safety inornata a stalin crete atta oleate tes 12 Safety standards Sample application large volumes medium sized volumes small volumes Sample loop eesssssssses complete filling of manual filling of partial filling of Sample preparation KTAprime plus User Manual 11 0026 44 Edition AA 199 200 Self test Set Parameters menus Spring tension Starting a run Storage Stored method COPYING ME AE seers N A hah AEN A UN Rie A ESE 80 editing running a Superloop applying samples with Switch valve System components ESCH OU OM Secssecsessszes cover sszescat snzsraecasasststesneracitectcaee eeastacstattnsldacssthmaset aE 156 location System control System flow path System self test T TrouDIESHOOUNG Lassen aE E AE E a A E R a E EAAS 143 U UV optical unit installing
52. adjust the bottom of the spring hold it near the lower end and lift or prise the bottom of the spring out of the hole e To increase the tension turn the spring counter clockwise e To decrease the tension turn the spring clockwise 10 13 Removing trapped air bubbles If there are large amounts of air in the tubing or if you suspect air in the pump purge the flow path as described in section 5 2 2 10 14 Restart after power failure If the power to the system is interrupted it automatically restarts when power is restored and displays the main operating menu All set values and the data from the latest run are retained and the lamp is switched on AKTAprime plus User Manual 11 0026 44 Edition AA Reference information 11 Reference information 11 1 System description AkTAprime plus is a compact separation unit comprising components for fluid handling and for measuring UV absorption conductivity and PH optional This section gives a brief description of the system and its components It also describes optional components that may be connected to the system A built in power control board supplies the components with power and a controller handles the communication between the components via an internal high speed network AkTAprime plus User Manual 11 0026 44 Edition AA 155 Reference information es 11 1 1 Components description
53. ap SP XL 5 ml 5x5m 17 5161 01 HiPrep 16 10 Q XL 1x20ml 17 5092 01 HiPrep 16 10 SP XL 1x20ml 17 5093 01 HiPrep 16 10 CM FF 1x20ml 17 5091 01 HiPrep 16 10 DEAE FF 1x20ml 17 5090 01 HiPrep 16 10 Q FF 1x20ml 17 5190 01 HiPrep 16 10 ANX FF high sub 1x20ml 17 5191 01 HiPrep 16 10 SP FF 1x20ml 17 5192 01 SSS 188 KTAprime plus User Manual 11 0026 44 Edition AA Reference information es Buffer Exchange Desalting Columns Column name Pack size Code no HiTrap Desalting 5 ml 5x5ml 17 1408 01 HiTrap Desalting 5 ml 100 x 5 ml 11 0003 29 HiPrep 26 10 Desalting 1x53ml 17 5087 01 HiPrep 26 10 Desalting 4x53 ml 17 5087 02 1 Pack size available on special order Size Exclusion Gel filtration Columns Column name Pack size Code no HiPrep 16 60 Sephacryl S 100 HR 1x 120m 17 1165 01 HiPrep 26 60 Sephacryl S 100 HR 1x 320m 17 1194 01 HiPrep 16 60 Sephacryl S 200 HR 1x120m 17 1166 01 HiPrep 26 60 Sephacryl S 200 HR 1x 320m 17 1195 01 HiPrep 16 60 Sephacryl S 300 HR 1x120m 17 1167 01 HiPrep 26 60 Sephacryl S 300 HR 1x 320m 17 1196 01 Chelating Columns Column name Pack size Code no HiTrap Chelating HP 1 ml 5x1ml 17 0408 01 HiTrap Chelating HP 5 ml 1x5ml 17 0409 01 HiTrap Chelating HP 5 ml 5x5ml 17 0409 03 HiTrap Chelating HP 5 ml 100 x 5 mlt 17 0409 05 1 Pack size available on special order Affinity Columns Column name Pack size Code no HiTrap Protein A HP 1 ml 5x1im 17 0402 01 HiTrap Prote
54. ay cause undesired operation AkTAprime plus User Manual 11 0026 44 Edition AA 185 Reference information 11 11 3 3 Wetted materials The wetted materials in AKTAprime plus are listed below XI XI X x x x x x x DIWDJaD ZWIS PjOD ssojH ZNO aslyddos 3dMWHN Aoj 6 3 X x follo 3d dd 34193 3413 d34 J4ld 33d winjuDyL auajfyjahjod yyBiam Jojnoajow yfy o4N Id MWHN aualAujaoson losojyoaua Auya 34193 aua AujaosJonyosja aua Ayys 3413 Jauufjodos auajfdosdauajAuyeosonysad 34 aua Aujaoson jos a Ajod 341d aUOJayJayjaJayyohjod 433d XI XI XI XI XI XI XI XI XI X XI x auajfidosdfijod dd auauyahjod Jd Jaqqniosonyiad W143 JOJD9UUO SUOIUN s4 14 JO U buign xX dooyadns IOPI MOjJ AJDA UOISJAAIP MOJA J NDA YMS NDA Uoo JANDA png JOSU S JNSS Jd XI JT XIW 40 9a 09 UONDD14 XI JOJIUOW dund W 44 AKTAprime plus User Manual 11 0026 44 Edition AA 186 Reference information 11 4 Chemical resistance guide and chemical compati bility The chemical resistance of AKTAprime plus to some of the most commonly used chemicals in liquid chromatography is indicated in the table below The ratings are based on the following assumptions 1 The synergistic effects of the chemical mixtures have not been taken into account 2 Room temperature and limited over
55. ction are faulty Call GE Healthcare Tubes skipped 1 The spring tension may be insufficient Perform the actions described in section 10 12 Drop synchronization is not functioning 1 Check that the drop synchronization function is turned on See section 11 2 1 2 The drop sensor photocell located above the tube sensor is dirty Clean the photocell with a damp cloth 3 Make sure that the fraction collection tubing is properly connected to the delivery arm See section 8 4 5 10 10 Buffer valve and injection valve Fault Action The valve is switching to wrong position The valve parts may have been incorrectly reassembled after replacement 1 Check that the distribution plate marking i o buffer valve or 3 injection valve is horizontal External leakage 1 Check the tubing connections Tighten or replace if required Internal leakage Internal leakage can be detected at the small hole on the underside of the valve body 1 Internal parts may be worn Change channel plate and distribution plate according to section 9 13 High back pressure 1 Perform cleaning in place by flushing the system with detergent 1 Change channel plate and distribution plate according to section 9 13 AkTAprime plus User Manual 11 0026 44 Edition AA 149 Troubleshooting es 10 11 Error messages If the suggested actions do not
56. de slope is out of range Check buffers and recalibrate slope lt 70 or gt 110 2 Clean the pH electrode and recalibrate 93 pH _cal failed 3 If the message remains call service check electrode 94 WARNING lt 1pH unit 1 ve ld a the pH of the buffers used during calibration must between cal_buff 1 amp 2 E LEOS Prune 95 Temp cal failed 1 Check that the conductivity cell is connected Recalibrate check cond cell 2 The measured temperature value differs from the reference value by more than 5 C or the actual temperature is lower than 8 C Recalibrate 97 WARNING pH scale 1 The difference between the zero level and full scale must be at least 1 pH 0 100 lt pH unit unit Increase the span between zero and full scale settings See section Viv2 A 98 Cal failed Cell 1 Conductivity cell constant is out of range constant not 0 1 300 2 Wrong solution used during calibration Use 1 00 M NaCl and recalibrate 3 Air in conductivity cell during calibration Flush the flow cell with calibration solution and recalibrate 4 Dirty conductivity cell Clean the flow cell and recalibrate If the problem remains change the conductivity cell 152 AKTAprime plus User Manual 11 0026 44 Edition AA Troubleshooting eE Messages Action 99 ERROR Out of 1 Maximum number of breakpoints in memory is 600 Delete a method to get method memory more memory ERROR key 1 Akey was pressed durin
57. e Hoffman La Roche Inc 2005 General Electric Company All rights reserved Amersham Biosciences AB a General Electric company going to market as GE Healthcare GE Healthcare Amersham Biosciences AB Bjorkgatan 30 751 84 Uppsala Sweden GE Healthcare Amersham Biosciences Europe GmbH Munzinger Strasse 9 D 79111 Freiburg Germany GE Healthcare Amersham Biosciences UK Ltd Amersham Place Little Chalfont Buckinghamshire HP7 9NA UK GE Healthcare Amersham Biosciences Corp 800 Centennial Avenue P O Box 1327 Piscataway NJ 08855 1327 USA GE Healthcare Amersham Biosciences KK Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 169 0073 Japan Asia Paci c Tel 852 2811 8693 Fax 852 2811 5251 Australasia Tel 61 2 9899 0999 Fax 61 2 9899 7511 Austria Tel 01 57606 1619 Fax 01 57606 1627 Belgium Tel 0800 73 888 Fax 03 272 1637 Canada Tel 800 463 5800 Fax 800 567 1008 Central East amp South East Europe Tel 43 1 982 3826 Fax 43 1 985 8327 Denmark Tel 45 16 2400 Fax 45 16 2424 e Finland amp Baltics Tel 358 0 9 512 39 40 Fax 358 0 9 512 39 439 e France Tel 01 69 35 67 00 Fax 01 69 41 96 77 Germany Tel 0761 4903 490 Fax 0761 4903 405 e Italy Tel 02 27322 1 Fax 02 27302 212 e Japan Tel 81 3 5331 9336 Fax 81 3 5331 9370 Latin America Tel 55 11 3933 7300 Fax 55 11 3933 7304 Middle East amp Africa Tel 30 210 9600 687 Fax 30 210 9600 693 Netherlands Tel 0165 580 4
58. eck valves next to the outlet upper port 2 Fasten the check valve using the screwdriver 3 Reinstall the connection part according to section 9 14 2 Installing the connection part 132 AkTAprime plus User Manual 11 0026 44 Edition AA Maintenance 9 E 9 17 Replacing mixer chamber WARNING When using hazardous chemicals make sure that the entire system flow path has been flushed thoroughly with distilled water before maintenance 1 Make sure the pump is stopped 2 Place the buffer bottles lower than the mixer to prevent draining and then remove the inlet and outlet tubing 3 Open the chamber lock holding the mixer chamber A spring is securing the chamber in position when the lock is opened 4 Pull out the mixer chamber gently 5 Move the stop plug to the right hand inlet of the new mixer chamber 6 Insert the new mixer chamber and close the lock 7 Replace the inlet and outlet tubing SSS AkTAprime plus User Manual 11 0026 44 Edition AA 133 9 Maintenance 134 9 18 Cleaning the pH electrode optional Note The pH electrode has a limited lifetime and should be replaced every six months or when the response time is slow or the slope is out of range lt 80 WARNING NaOH is injurious to health Avoid spillage Use one of the following procedures to clean the electrode to improve the response e Salt deposits Dissolve the deposit by immersing the electrode first in 0 1 M HCI then
59. ectly prepared Prepare a new calibration solution and recalibrate the conductivity cell Ghost peaks appear in the gradient profile 1 A charged sample has been detected e g protein 2 Air bubbles are passing through the flow cell Check for loose tubing connections If necessary use a flow restrictor after the conductivity cell Non linear gradients or slow response to B changes 1 Check that the tubing has been washed properly and that the pump is operating 2 Change to a smaller mixer volume 146 AKTAprime plus User Manual 11 0026 44 Edition AA Troubleshooting 10 6 pH curve optional Fault Action Incorrect unstable pH reading 1 Check that the electrode cable is connected properly to rear of the system 2 Check that the pump operates properly 3 Check that the electrode is correctly inserted in the flow cell and if necessary hand tighten the nut 4 If airin the flow cell is suspected tap the flow cell carefully or tilt it to remove the air Alternatively flush the flow cell with buffer at 20 ml min for 30 s Use a flow restrictor after the pH electrode 5 Check that the pH electrode is not broken 6 Check that the pH electrode is calibrated Check the slope If it is outside the range 80 105 or if the asymmetry potential deviates more than 60 mV from 0 mV clean the pH electrode Recalibrate If the problem persists replace the pH electrode 8 Clean the pH electrode if
60. ector allows fixed volume fractionation or automatic peak fractionation Fraction marks make it easy to identify fractions and peaks Fast tube change minimizes spills between tubes eliminating it entirely below flow rates of 5 ml min Drop synchronization eliminates sample loss during tube change The flow diversion valve mounted on the fraction collector is used as follows e PortIN Connected to the flow restrictor e Port NO normally open Connected to waste e Port NC normally closed Connected to the tubing holder fitted on the delivery arm on the fraction collector G 162 KTAprime plus User Manual 11 0026 44 Edition AA Reference information 11 1 2 Electrical connections All electrical connections for AKTAprime plus are located at the rear of the system Mains cable D l Ground pH Probe Drop Sensor Frac Vale reorder O o w rs232 o Conductivity Flow Cell LJ O Gp Om Soo eaag eee ars oa ae a oe om fae a Soon ooo eeeSeac Mains vamp T EEO fa C Mains switch From mains outlet One mains input is required for the system Communication cables RS 232 to computer From pH electrode To flow diversion From conductivity valve flow cell From fraction collector ph Ground pHfprobe From optical unit
61. es 2 Repeat the ultrasonic bath with distilled water 9 16 5 Replacing the check valve O rings Liquid appearing at a banjo fitting might indicate that a check valve O ring is damaged It might also cause reduced flow or pressure fluctuation Carefully replace both O rings on the check valve Oring with new ones if you suspect that an O ring is damaged 9 16 6 Installing the check valves If cleaning of the check valve as described in section 9 16 4 Cleaning the check valves off line does not correct the fault replace the check valve with a new one Note Make sure that the check valves are installed in their correct positions By mistake an inlet check valve can be installed incorrectly in an outlet check valve position and vice versa The check valves are of four different types Inlet check valve short 1 pc Inlet check valve long 2 pcs Outlet check valve short 2 pcs Outlet check valve long 1 pc AkTAprime plus User Manual 11 0026 44 Edition AA 131 9 Maintenance ay The inlet check valves have line shaped holes The outlet check valves have round holes Inlet check valve long Line shaped pe o_O Inlet check valve short 1 Outlet check valve long UZIT m Poa holes Outlet check valve short WM To install a check valve 1 Carefully insert the check valve fully Note Make sure that the inlet check valves are installed next to the inlet lower port on the connection part and the outlet ch
62. g self test or is faulty 2 Switch off the system 3 Switch on the system ERROR Number 102 104 1 Switch off the system 2 Check all connections ERROR Number 109 113 3 Switch on the system ERROR Number 119 121 Exc x y in ab c Switch off the system 2 Check all connections Exc DIV 0 in ab c 3 Switch on the system Exc instr in ab c Exc address in ab c AkTAprime plus User Manual 11 0026 44 Edition AA 153 Troubleshooting 154 10 12 Adjusting the spring tension of the delivery arm Incorrect spring tension can cause the fraction collector to skip tubes The effect is greater as the arm moves towards the centre Spring tension is temperature sensitive Low temperature for example in a cold room reduces the spring tension so it may be necessary to adjust the tension 1 Remove the arm bracket from the stand 2 Dismantle the delivery arm from the bracket 3 The top of the spring is fastened in one of two holes in the top of the arm bracket Looking at the arm bracket from the front of the unit moving the spring from the right hand to left hand hole increases the tension and conversely moving from the left hand to the right hand hole decreases the tension Hold the spring near the top and pull or prise it down and out of the top hole Insert the spring in the other hole 4 The bottom of the spring is fastened in one of four holes equally spaced 1 4 turn apart To
63. gth is shown Checking lamp run time The lamp run time can be checked to determine the need for lamp replacement Run times for both Hg and Zn lamps are monitored 1 Select menu Check and press OK 2 Select sub menu Check Lamp Run Time Checking lamp intensity The lamp intensity can be checked to determine the status of the lamp used 1 Select menu Check and press OK 2 Select sub menu Check Lamp Intensity 173 Reference information Check Pump Run Time 246h Check Pumped Volume 3567 ml Check Tube Shifts 3592 Check Valve Shifts BV 642 IV 348 Check Recorder 174 Checking pump run time The pump run time can be checked to determine the need for maintenance 1 Select menu Check and press OK 2 Select sub menu Check Pump Run Time Checking pumped volume The volume delivered by the pump can be checked to determine the need for maintenance 1 Select menu Check and press OK 2 Select sub menu Check Pumped Volume Checking tube shifts The number of tube shifts done by the fraction collector can be checked to determine the need for maintenance 1 Select menu Check and press OK 2 Select sub menu Check Tube Shifts Checking valve shifts The number of shifts done by the buffer valve and the injection valve can be checked to determine the need for maintenance 1 Select menu Check and press OK 2 Select sub menu Check Valve Shifts Checking the rec
64. he mini DIN connector to socket Recorder e The MC connector to socket Rec On off The left hand pin in the socket is the ON OFF signal and the right hand pin is signal ground 00o KTAprime plus User Manual 11 0026 44 Edition AA Installing and modifying components 8 6 Connect the pin connectors at the other Input 2 signal Input 2 ground end of the cable to the signal input plugs supplied with the recorder Note The signal cable is delivered with T Chassis protective covers on each wire Do ground not remove the protective covers from unused connections as this an may disturb the measurement Input 1 signal Input 1 ground 7 Connect the plugs to the signal inputs on the recorder Pin designations for the signals and colors on the corresponding cable wire are as follows Pin no Signal Range 1 Brown Signal 1 0 1V 2 Red Signal 1 ground OV 3 Orange Signal 2 0 1V 4 Yellow Signal 2 ground OV 5 Green Signal 3 0 1V 6 Black thin Signal 3 ground OV 7 Black thick Chassis ground OV 1 The signals have the following default parameter settings Signal 1 UV absorbance Signal 2 B Signal 3 Conductivity These settings can be changed see section 8 6 5 Setting analog outputs 8 Connect the D sub connector to the D sub socket in the recorder Pin
65. he tube sensor rests P against tube 1 When the fraction collector is ras started the bowl moves to the correct position to collect the first fraction in tube 1 Sensor control Check that the sensor is in the correct position for the tube size The eluent tubing should be over the centre of the collection tube Use the red sensor control to position the tube holder AkTAprime plus User Manual 11 0026 44 Edition AA 107 ED Installing and modifying components A 8 4 5 Connecting tubing Select the tubing with the required inner diameter 0 75 mm To change the tubing follow steps 1 5 WARNING When using hazardous chemicals avoid spillage during fraction collection and when the delivery arm is moved out 1 Fita41cmlong tubing by lifting out the tubing holder from the delivery arm loosening the nut and then inserting the tubing Note Check that the tubing end is cut leaving a straight edge Note The tubing must be long enough to ensure free movement of the delivery arm Tubing holder 2 Place the tubing holder over the length guide small hole in the delivery arm push the tubing down to the bottom of the guide and tighten the nut This ensures that the correct length of tubing is exposed 3 Re install the tubing holder into the delivery arm 4 Set the red sensor control to position the Sensor Control tubing over the centre of the collection tube 5 Connect the other end of the tubing to por
66. heck that inlet or outlet tubings are not clogged Replace if necessary 3 A check valve in the pump might be clogged or damaged Remove the check valve according to section 9 14 Clean the check valve in an ultrasonic bath If the leakage persists replace the assembly Irregular flow noisy baseline signal Air bubbles passing through or trapped in the pump irregular pressure trace 1 Check that there is sufficient eluent in the reservoirs 2 Check all connections for leakage 3 Use degassed buffers 4 Remove any air bubbles according to section 10 13 Blockage or partial blockage of the flow path 1 Flush the flow path to clear the blockage 2 If necessary replace the tubing 3 Check the inlet tubing filter 4 A check valve in the pump might be clogged or damaged Remove the check valve according to section 9 14 Clean the check valve in an ultrasonic bath If the leakage persists replace the assembly 144 AkTAprime plus User Manual 11 0026 44 Edition AA Troubleshooting 10 4 UV curve Fault Action Noisy UV signal signal drift or instability 1 Select menu Check Autozero to check the autozero absorbance value If the value is between 1 5 and 2 there may be air bubbles in the flow cell or the wrong buffer system is in use 2 Wrong filter for the lamp is being used Check that the lamp is in the proper position and that the correct filter is used 3 The buffer may be impure Check if the signal
67. in A HP 1 ml 2xim 17 0402 03 HiTrap Protein A HP 5 ml 1x5m 17 0403 01 HiTrap Protein G HP 1 ml 5x1im 17 0404 01 HiTrap Protein G HP 1 ml 2xim 17 0404 03 HiTrap Protein G HP 5 ml 1x5m 17 0405 01 HiTrap Heparin HP 1 ml 5x1im 17 0406 01 HiTrap Heparin HP 5 ml 1x5m 17 0407 01 HiPrep 16 10 Heparin FF 1x20ml 17 5189 01 HiTrap rProtein A HP 1 ml 5x1im 17 5079 01 HiTrap rProtein A HP 1 ml 2xim 17 5079 02 as AkTAprime plus User Manual 11 0026 44 Edition AA 189 Reference information Es Column name Pack size Code no HiTrap rProtein A HP 5 ml 1x5ml 17 5080 01 HiTrap Blue HP 1 ml 5x1ml 17 0412 01 HiTrap Blue HP 5 ml 1x5ml 17 0413 01 HiTrap NHS activated HP 1 ml 5x1ml 17 0716 01 HiTrap NHS activated HP 5 ml 1x5ml 17 0717 01 HiTrap NHS activated HP 1 ml 180 x 1 ml 17 0716 30 HiTrap IgM Purification HP 1 ml 5x1ml 17 5110 01 HiTrap IgY Purification HP 5 ml 1x5ml 17 5111 01 HiTrap Streptavidin HP 1 ml 5x1ml 17 5112 01 HisTrap HP 1 ml 5x1ml 17 5247 01 HisTrap HP 1 ml 100 x 1 ml 17 5247 05 HisTrap HP 5 ml 1x5ml 17 5248 01 HisTrap HP 5 ml 5x5ml 17 5248 02 HisTrap HP 5 ml 100 x 5 mlt 17 5248 05 HisTrap FF 1 ml 5x1ml 17 5319 01 HisTrap FF 1 ml 100 x 1 ml 17 5319 02 HisTrap FF 5 ml 5x5ml 17 5255 01 HisTrap FF 5 ml 100 x 5 mlt 17 5255 02 HisPrep FF 16 10 1x20ml 17 5256 01 HisTrap FF crude 1 ml 5x1ml 11 0004 58 HisTrap FF crude 1 ml 100 x 1 ml 11 0004 59 HisTrap FF crude 5 ml 5x5ml 17 52
68. insert the electrode If so loosen the inlet connection while inserting the electrode to allow the liquid to run out from the flow cell Remember to re tighten the connector Electrode Note Ifthe electrode is not fully inserted the system will leak and a dead volume will occur in the holder Flow cell Flow cell 4 Connect the pH electrode cable to the socket holder pH Probe on the rear of the system AkTAprime plus User Manual 11 0026 44 Edition AA 109 ED Installing and modifying components MC D sub Pin connetor E 110 Mini DIN 8 6 Recorder REC 112 AkTAprime plus can be delivered with a recorder REC 112 for simpler data presentation This section describes have to install and use the recorder 8 6 1 Electrical connections 1 Make sure that the mains power switch is in the OFF position Insert the correct fuses into the fuse caps Fuse caps 2 pcs e For 100 120 V operation use the 250 mA fuses supplied e For 220 240 V operation use the 125 mA fuses supplied Fit the fuse caps into the fuse holder sockets on the left side of the recorder Make sure that the mains voltage selector at the rear of the recorder is set to the mains voltage of the laboratory CAUTION The mains voltage selector must be set to the mains voltage of the laboratory If not electronics might be damaged Connect the supplied signal cable to AKTAprime plus as follows e T
69. ion Connector A1 A B ut and ferrule for o d 3 16 a tubing blue ferrule AB Nut and ferrule for o d 1 8 tubing yellow ferrule iii W1 W3 G1 G6 F1 Fingertight connector for o d 1 16 tubing Union male male Union 1 16 male 1 16 male Stop plug Stop plug 1 16 AKTAprime plus User Manual 11 0026 44 Edition AA 165 Reference information 166 11 1 4 Optional components pH flow cell with electrode The pH electrode is of the sealed combination double junction type It contains a sealed Ag AgCl reference which cannot be refilled an internal electrolyte bridge of 4 M KCI saturated with Ag AgCl an outer electrolyte bridge of 1 M KNO3z an annular ceramic reference junction and a low profile pH membrane The pH electrode is delivered in a transparent cover The flow cell is made of titanium It should not be used with any other pH electrode Superloop Superloop allows introduction of larger sample volumes into a pressurized fluid system It is used together with the injection valve and replaces a simple sample loop Superloop consists of a movable seal in a glass tube The seal divides the tube into two separate chambers Depending on the flow direction the seal moves towards either end piece of the glass tube Superloop is available in three sizes 10 50 and 150 ml allowing application of 1 10 1 50
70. ion and press OK calibration Set Mix Chamber Vol 3 Select sub menu Set Mix Chamber Vol The current chamber volume is 2 ml shown Press OK to change the volume Set Mix Chamber Vol 4 Change the setting as desired and press OK Possible values are 0 6 2 0 5 0 2 ml 2 0 and 12 0 ml SSS 172 KTAprime plus User Manual 11 0026 44 Edition AA Check Communication PC App Connected Check Communication PC Driver Connected Check Communication Not Connected Check Autozero AZ 0 00006AU Check Lamp Run Time Hg 1482h Zn 430h Check Lamp Intensity R 215 5 S 214 4mV AKTAprime plus User Manual 11 0026 44 Edition AA Reference information 11 2 2 Check menus Checking communication The communication between the AKTAprime plus system and a computer can be checked 1 Select menu Check and press OK 2 The system immediately checks the communication with the computer PC App Connected The communication is OK PC Driver Connected PrimeView software is not properly installed Not Connected The serial cable between the system and the computer is not properly connected or PrimeView software is not installed on the computer Checking autozero level The module internal absorbance value for autozero can be checked to test the consistency of buffers 1 Select menu Check and press OK 2 Select sub menu Check Autozero The autozero absorbance value for the used wavelen
71. is displayed as default Enter the temperature shown on the thermometer and press OK 139 9 Maintenance Set Parameters Setup and calibration Setup pH Calibrate pH 7 00 12 00 Calibrate pH Buffer 1 Calibrate pH Buffer 1 7 00 Please wait 140 9 20 5 Calibrating the pH electrode optional A good laboratory routine is to calibrate the pH measurement once a day when the electrode is replaced or if the ambient temperature changes The pH electrode is calibrated using standard buffer solutions in a two point calibration The two buffer solutions can have any pH value as long as the difference between them is at least 1 pH unit The calibration procedure can be done with the pH electrode either fitted in or removed from the flow cell Calibrating with the electrode outside the flow cell When calibrating the electrode out of the flow cell and changing from one buffer to another rinse the electrode tip with distilled water and dab it carefully with a soft tissue to absorb the remaining water Do NOT wipe the electrode as this may charge it and give unstable readings The steps below describe the procedure used with the electrode removed from the flow cell 1 Remove the pH electrode from the flow cell and immerse the electrode in the first standard buffer solution normally pH 7 0 2 From the main menu select menu Set Parameters by pressing the up or the d
72. is still noisy with water 4 There may be air in the flow cell Check that the flow restrictor generates a back pressure of 0 2 0 05 MPa Replace it if this iS not within the limits If there is a lot of air in the water degas the buffer before use Check the connections of the optical unit Clean the UV flow cell see sections 9 9 and 9 10 oN DWM Locking nut in optical unit not properly tightened Turn the locking nut to the stop position Ne Air bubbles trapped in the pump Refer to section 10 13 Ghost peaks Check that there is no air in the eluent If necessary degas the eluent 2 Clean the system in accordance with section 9 3 Clean the column in accordance with the column instructions 4 Check that the mixer is functioning properly and that the correct chamber volume is being used 5 Unless you are using a low pressure column try using a flow restrictor FR 904 instead of FR 902 This generates a higher back pressure 0 4 MPa instead of 0 2 MPa Low sensitivity 1 Aging lamp Check the lamp and replace if necessary 2 Wrong lamp position Check that the lamp position and the wavelength used filter position fit together Error in external chart recorder 1 Check the recorder according to the manufacturer s instructions AKTAprime plus User Manual 11 0026 44 Edition AA 145 Troubleshooting 10 5 Conductivity curve Fault Action Incorrect or unstable reading
73. it in buffer overnight 3 Clogged liquid junction Refer to section 9 18 pH values vary with varied back 1 Replace the pH electrode pressure 10 7 Mixer Fault Action Leakage 1 Check the tubing connections Retighten or replace if necessary 2 Check the mixer chamber Replace if liquid has penetrated the mixer chamber walls and sealings 10 8 Pump Fault Action Irregular flow 1 Check the inlet and outlet tubings 2 Remove any air bubbles according to section 5 2 2 3 If still irregular flow calibrate the pump according to section 9 20 1 4 A check valve in the pump might be clogged or damaged Remove the check valve according to section 9 14 Clean the check valve in an ultrasonic bath If still irregular flow replace the assembly Leakage 1 An O ring in a check valve or in the connection part might be damaged Examine the O rings If necessary replace them according to the section 9 15 2 or 9 16 5 SEE 148 AkTAprime plus User Manual 11 0026 44 Edition AA Troubleshooting a 10 9 Fraction collector Fault Action No tube change 1 Press the feed tube key If the motor does not start and an error appears call GE Healthcare 2 Push the delivery arm out to a safety stop Press the feed tube key If the motor starts press the tube sensor together within 2 seconds The motor should stop without an error code reported If an errors appears check the connection in the arm If this is correct the sensor or sensor conne
74. ition AA 197 198 F Faults and actions Filling the buffer inlet tubing Filling the sample loop Final checks Fixed volume fractionation Flow diversion valve F F F F ow path ow path control ow rate range ow restrictor FR 902 column pressure pressure limits Fluid handling path Fraction collection Fraction collector installing troubleshooting Fractions collecting G e atlar StaMed PEE A EA EE E AEDE E E ok khakis oe ete Bat 21 H Histidin tagi roscoe sees ic aera ee A lk a E R 21 I FUJSCTION TU DOE E ie I la a le oe a od as 46 njection needle njection valve replacing parts troubleshooting nstallation nstallation procedure ntegrating peaks 30 L Liquid delivery M Main menu overview 40 Mains cable Maintenance Menu navigation Menu overview Method templates running a Mixer troubleshooting 148 Monitor Monitoring conductivity pH UV absorbance AkKTAprime plus User Manual 11 0026 44 Edition AA Index LAS o Operating data Ordering information P Portiqlifiling sss Tearen a a E E T Peak fractionation Periodic maintenance pH electrode installing Post run printing Power requirement Preparing the fraction collector Preparing the monitors Preparing the system for arun Preparing the tubing and column Pressure limits Pressure range
75. les to enter the flow cell as damage to the flow cell may occur 1 Connect a syringe to the inlet of the flow cell and squirt distilled water through the cell in small amounts Then fill the syringe with a 10 surface active detergent solution like Decon 90 Deconex 11 RBS 25 or equivalent and continue to squirt five more times 2 Leave the detergent solution in the flow cell for at least 20 minutes AKTAprime plus User Manual 11 0026 44 Edition AA Maintenance 9 a 3 Pump the remaining detergent solution through the flow cell 4 Rinse syringe and flush the flow cell with distilled water 10 ml 9 11 Cleaning the conductivity flow cell off line WARNING NaOH is injurious to health Avoid spillage If the conductivity measurements are not comparable to previous results the electrodes in the flow cell may be contaminated and require cleaning To clean the flow cell 1 Pump 15 mlof1MNaOH at 1 ml min through the flow cell either by using the system pump or a syringe 2 Leave for 15 minutes 3 Rinse thoroughly with degassed distilled water Note Ifthe flow cell is totally blocked the blockage can be removed using a needle or a wire with a diameter less than 0 8 mm 9 12 Replacing the conductivity cell The conductivity flow cell can be replaced when required Make sure the system is switched off before connecting disconnecting the cell from the rear of the system If the cell is replaced with a
76. leshooting Messages Action 69 ERROR Stop grad set HOLD or PAUSE 1 Set the system in HOLD or PAUSE or stop the gradient B 70 Lamp disconnected If not call service 1 Connect the lamp or call service 71 WARNING low light intensity Check the cables to the optical unit Check that lamp and filter position correspond Change lamp If the problem remains call service 72 Change lamp or call service 76 Change lamp or call service If used in cold room additional warm up might be needed If the problem remains change the lamp WNW N e N e If the problem remains call service straylight leaks in 73 WARNING Too much 1 Check that the filter wheel cover is closed Check that non transparent tubings are used at the UV flow cell inlet and outlet Check that the optical unit is not exposed to direct sunlight If the problem remains call service 77 WARNING Autozero out of range Autozero is not allowed on a level above 2 AU Check buffers Clean UV flow cell 78 ERROR Over pressure e N PIP WNW Re program the method 79 ERROR Method corrupt in eeprom 1 Check which method is corrupted erased 2 Ifthe problem remains call service 80 ERROR Reading from eeprom 1 Call service 81 ERROR Writing to eeprom 1 Call service 82 ERROR Parameter fail in method 1
77. lists faults observed with specific monitor measurements and the specific components The faults are listed as follows Type Page USE e 2s Ac conse nce E EE E E EE onsen EE EEEE E 144 UM CUNE igu d ates tines dee Re edge soe S 145 GONGUCEVITY CURVE is cede eee daear Ce Sie Let eee eae ce bade ed 146 PH curve optiondl 0 ccc cence cece RESE IVE een es 147 RFESSULE CUVE erei aan aaa E ahaa serena pave oon wine nae an ute O DEO ae 144 210 0 ease ae eee EN ae ea ee eR 148 MIRON tie ca e s aye ett iol E EE EOE mie E pla nga Glee sige nwa plese 148 Fraction collecto si oct ds ceedte ening oy He dened g Metab io NETET avis ed ees 149 Buffer valve and injection valve 2 ccc cence nee e ence enes 149 If the suggested actions do not correct the fault call GE Healthcare AKTAprime plus When contacting GE Healthcare for support state the program version of the V 3 00 system which is shown for a few seconds during start up WARNING The system should not be opened by the user It contains high voltage circuits that can deliver a lethal electric shock AkTAprime plus User Manual 11 0026 44 Edition AA 143 Troubleshooting 10 2 System Fault Action No text on the front display 1 Check that the mains cable is connected and that the power is turned on 10 3 Pressure curve Fault Action Pressure limit exceeded inaccurate 1 Calibrate the pressure monitor reading 2 C
78. lue in the display with the up and down buttons so that it corresponds to the known pH value of the second buffer solution Press OK 12 After the calibration with buffer 2 the system automatically Voltage mV The calibration curve enters the Calibrated Electrode 500 mV shows the relationship menu This menu shows the Asymmetry between pH and the i potential output signal from the slope of the calibration curve atpH7 monitor in mV where 100 corresponds to 59 16 mV per pH step at 25 C The asymmetry potential at pH 7 is shown as a mV value Press Esc repeatedly to return to the 500 mV Set Parameters menu 13 Before use rinse the electrode using distilled water A new electrode typically has a slope of 95 102 and an asymmetry potential within 30 mV As the electrode ages the slope decreases and the asymmetry potential increases As arule when an electrode has an asymmetry potential outside 60 mV and a slope lower than 80 and no improvement can be made by cleaning the electrode should be changed An electrode is still usable at lower slopes and higher asymmetry potentials but the response will be slower and the accuracy diminished Calibrating with the electrode in the flow cell When calibrating with the electrode fitted in the flow cell follow the above procedure but let at least 30 35 ml with 2 ml mixer of standard buffer solution be pumped through the system to stabilize pH Leave the
79. m 194 KTAprime plus User Manual 11 0026 44 Edition AA Reference information Sy Item Quant A C Code no pack Fingertight connector 1 16 10 A 18 1112 55 ar PEEK tubing o d 1 16 Stop plug 1 16 PEEK 5 A 18 1112 52 Stop plug 5 16 PEEK 5 A 18 1112 50 Tubing Teflon tubing i d 2 9 mm o d 3 16 IN 3m A 18 1112 47 PEEK tubing i d 0 50 mm 2m A 18 1113 68 o d 1 16 PEEK tubing i d 0 75 mm 2m A 18 1112 53 o d 1 16 G PEEK tubing i d 1 0 mm o d 1 16 W 2m A 18 1115 83 Sample tubing kit 1 A 18 1115 77 Purge kit 1 A 18 1153 28 Miscellaneous Inlet filter assembly 2 A 18 1113 15 Inlet filter set 10 18 1114 42 On line filter 1 A 18 1112 44 On line filter kit 10 C 18 1027 11 Screw lid GL45 incl cap membrane 1 A 11 0004 10 Flow restrictor FR 902 1 A 18 1121 35 Flow restrictor FR 904 1 A 18 1119 63 Cramp for column holder KTAprime plus 1 A 18 1142 71 Column holder for one column short 1 A 18 1113 17 Column holder for one column long 1 A 18 1126 32 Tubing cutter 1 A 18 1112 46 U wrench M6 1 A 19 7481 01 U wrench 1 4 1 A 18 1112 45 Hex wrench 2 5 mm 1 A 19 4442 01 Chart recorder REC 112 2 channel 1 A 18 1132 33 Computer laptop 1 A 18 3535 35 KTAprime plus User Manual 11 0026 44 Edition AA 195 Reference information 196 Item Quant A C Code no pack User Documentation AKTAprime plus User Manual AKTAprime plus Cue Cards Related li
80. n part O ring x7 9 16 Cleaning and replacing check valves in the pump Faulty operation of the check valves is usually indicated by irregular flow very low flow or unstable pressure traces Probable causes are air dirt or a damage in a check valve preventing it from closing to seal and hold the pressure Liquid appearing at a banjo fitting might indicate that a check valve O ring is damaged 9 16 1 Required spare parts e Check valve there are four different check valves See Ordering information for code nos e O ring kit see Ordering information for code no 9 16 2 Cleaning the check valves in place Try to clean the check valves in place by pumping 100 ethanol for approximately 10 min If this does not correct the problem follow the instructions below to remove and then clean the valves If necessary a check valve or O rings might need to be replaced AKTAprime plus User Manual 11 0026 44 Edition AA Maintenance 9 9 16 3 Removing the check valves If the condition of the check valve is not improved by in place cleaning remove it as follows 1 Remove the connection part according to section 9 14 1 Removing the connection part 2 The check valves in the connection part are locked in position with banjo fittings Remove the check valves using the screwdriver Check valve 9 16 4 Cleaning the check valves off line 1 Immerse the check valves in 100 ethanol and place in an ultrasonic bath for 5 10 minut
81. nal comes from the same point in the lamp as the signal measuring the sample thus assuring a stable baseline by eliminating the effects of variations in lamp intensity Optical Unit Inlet Filter Lens Beam splitter Flow cell Photodetector _ Outlet Photodetector vr Vs System housing Front panel Microprocessor High voltage power supply AkTAprime plus User Manual 11 0026 44 Edition AA 159 Reference information The Hg lamp emits light only at certain wavelengths It does not emit light at 280 nm so for this wavelength the light is converted at a fluorescent surface before it passes the filter On the lamp housing there is a special exit for 280 nm light which means that the lamp position needs to be changed when working with this wavelength For 214 nm wavelength a Zn lamp is used This lamp is larger than the Hg lamp and is therefore mounted in a larger lamp housing The lamp connectors are keyed to inform the monitor software which lamp type is connected Conductivity flow cell The flow cell has two cylindrical titanium electrodes positioned in the flow path of the cell An alternating voltage is applied between the electrodes and the resulting current is measured and used to calculate the conductivity of the eluent The monitor controls the AC frequency and increases it with increasing
82. on 1 is directly above position 1 in the tube guide Push the flexible bowl out at each rib and snap the holder under the top of the rib SE AkTAprime plus User Manual 11 0026 44 Edition AA 105 ED Installing and modifying components n Do not force the holder into place as this may damage the lip The surface of the holder should be level 8 4 2 Mounting the tube rack 1 Gently move the delivery arm out to the second stop 2 Place the rack over the central spindle and pull the spring loaded drive sleeve out so that the rack comes to rest Drive sleeve 8 4 3 Inserting the collection tubes Insert the sufficient collection tubes into the rack starting in position 1 pushing each one down as far as it will go All the tubes must be of the same length and diameter and there should be no empty spaces in the sequence SSS 106 AkTAprime plus User Manual 11 0026 44 Edition AA Installing and modifying components 8 ae 8 4 4 Adjusting the delivery arm 1 Lower the arm and allow it to move so that the tube sensor touches the collection tubes of the outer track Tube sensor 2 Adjust the arm bracket so that the bottom of the tube sensor is about 5 mm below the top ee of the tubes The tubes should always be below the horizontal mark on the tube sensor 3 Lock the arm bracket at this height with the lock knob 4 Rotate the rack counter clockwise by hand until the rear half of t
83. optional or contact GE Healthcare for lamp replacement AKTAprime plus User Manual 11 0026 44 Edition AA Maintenance ex ae 9 5 2 Checking lamp run time Check Lamp Run Time 1 Select menu Check and press OK Hg 2300h Zn 340h 2 Select menu Check Lamp Run Time e The lifetime of a Hg lamp at 254 nm is typically 7000 hours in room temperature in coldroom typically 2000 h e The lifetime of a Hg lamp at 280 nm is typically 3500 hours in room temperature e The lifetime of a Zn lamp is typically 2000 hours in room temperature When necessary replace the lamp assembly according to section 8 2 5 Changing the lamp assembly optional or contact GE Healthcare for lamp replacement 9 5 3 Checking autozero The internal absorbance value for autozero can be checked to test the consistency of buffers 1 Select menu Check and press OK Check Autozero 2 Select menu Check Autozero The autozero absorbance value for the AZ 0 0001 AU wavelength used is shown 9 6 Checking the pump 9 6 1 Checking pump run time 1 Select menu Check and press OK Check Pump Run Time 2 Select menu Check Pump Run Time 00014h The lifetime of the wearing parts glass tube piston sealings and O rings is typically 2000 hours at normal use When necessary contact GE Healthcare for replacement 9 6 2 Checking pumped volume 1 Select menu Check and press OK Check Pumped volume 2 Select menu Check Pum
84. order The function of a connected chart recorder can be tested 1 Select menu Check and press OK 2 Select sub menu Check Recorder 3 Press OK to start the test The test will ramp the signal on each channel up to 1 V and then immediately 1v decrease the signal back to 0 V The test is run continuously Compare the diagram of the chart recorder with the figure Signal to recorder 4 Stop the test by pressing OK or Esc AKTAprime plus User Manual 11 0026 44 Edition AA Check Service Mode Telephone Service 012345678901 Contract number 01234567801 Serial Number 0123456 YM 012345 KTAprime V3 00 Date of maintenance DD MMMM YYYY Buzzer Test Reference information Checking service mode Service information relevant to the module can be checked Information may not be available in all menus 1 2 Select main menu Check and press OK Select menu Check Service Mode and press OK The service telephone number is displayed Press OK The service contract number is displayed Press OK The module serial number is displayed Press OK The system name and software version are displayed Press OK The date of the last service is displayed Press OK A test of the system buzzer is performed Press OK AkTAprime plus User Manual 11 0026 44 Edition AA 175 Reference information _ 11 2 3 Templates Menu overview Ap
85. ore disassembling the pump move the input liquid bottle below the level of the pump to prevent siphoning 1 Turn off the system with the mains power switch 2 Disconnect the inlet and the outlet tubing IT from the connection part 3 Release the pump from the system _ 4 Unscrew the two attachment screws using a 4mm hex wrench 5 Remove the connection part l L J Connection part Screws 9 14 2 Installing the connection part 1 Wipe the back of the connection part and the other parts behind the connection part with a clean cloth 2 Check that none of the seven O rings on the rear side of the connection part has come loose 3 Fit the connection part in position Fasten the two attachment screws using the 4 mm hex wrench 4 Reconnect the inlet and the outlet tubing 5 Purge the pump carefully and check that the fault is corrected See section 5 2 2 Purging pump and inlet tubing SSS AkTAprime plus User Manual 11 0026 44 Edition AA 129 9 Maintenance 130 9 15 Replacing O rings in the pump 9 15 1 Required spare parts e O ring kit see Ordering information for code no 9 15 2 Replacing an O ring To replace an O ring 1 Remove the connection part according to section 9 14 1 Removing the connection part 2 Remove the faulty O ring carefully to avoid making scratches on the connection part 3 Fit the new O ring in position 4 Reinstall the connection part according to section 9 14 2 Installing the connectio
86. ound in the Recorder REC 112 User Manual supplied AKTAprime plus User Manual 11 0026 44 Edition AA Maintenance ex 9 Maintenance This chapter contains a periodic maintenance schedule and instructions for maintenance replacing components and calibrations 9 1 Periodic maintenance Regular maintenance will help to keep your AKTAprime plus running smoothly Follow the recommendations in this chapter to keep the system in good working order WARNING Remove liquid or dirt from the system surface using a cloth and if necessary a mild cleaning agent WARNING Always disconnect the power supply before attempting to replace any item on the system during maintenance WARNING If there is a risk that spilt liquid have penetrated the casing of the instrument and come into contact with the electrical components immediately switch off the system and contact an authorised service technician WARNING When using hazardous chemicals take all suitable protective measures such as wearing protective glasses and gloves resistant to the chemicals used Follow local regulations and instructions for safe operation and maintenance of the system WARNING When using hazardous chemicals make sure that the entire system has been flushed thoroughly with bacteriostatic solution e g NaOH and distilled water before service and maintenance WARNING NaOH is injurious to health Avoid spillage WARNING Only spare parts
87. own button Press OK 3 Select menu Setup and calibration Press OK 4 Select menu Setup pH and press OK 5 Select menu Calibrate pH Current calibration values are displayed buffer 1 buffer 2 Buffer 1 fixed lower calibrated pH value Range 0 00 14 00 Buffer 2 fixed higher calibrated pH value Range 0 00 14 00 Note The values for buffer 1 and 2 must differ by at least 1 pH unit 6 Press OK to access the settings menu The order of calibration buffer 1 or buffer 2 is optional Press OK to start with buffer 1 or press the down button to start with buffer 2 In this example we start with buffer 1 7 This text disappears when the reading is stable and the following text is then shown Calibrate pH Buffer 1 7 00 7 00 AKTAprime plus User Manual 11 0026 44 Edition AA Calibrate pH Buffer 2 Calibrate pH Buffer 2 9 00 Please wait Calibrate pH Buffer 2 9 00 9 00 Calibrated Electrode Slope 98 5 9 5 mV Maintenance 9 8 Adjust the pH value in the display with the up and down buttons so that it corresponds to the known pH value of the first buffer solution Press OK 9 Atthe buffer 2 calibrating menu rinse the electrode tip with distilled water and then immerse the electrode in the second buffer solution e g pH 4 0 or 9 0 Then press OK 10 The text opposite disappears when the reading is stable and the text below is then shown 11 Adjust the pH va
88. pH Analogue Out Set pH Zero Level dH 13 00 3 00 RIEAN Set Press Analog Out Set pH Full Scale 1 00 MPa pH 13 00 Calibrate pH Calibr pH Buffer 1 Set pH Temp Comp Calibr pH Buffer 2 Set Show pH Calibrated Electrode on Slope 100 0 12 5 mV Set Cond Temp Comp Set Comd Ref Temp LS Setup pH Set Adjust Cond Setup Cond Set Adj Cell Const 4 Setup UV Set Show Cond Set Show UV on Set Lamp Run Time Setup Temp Set Averaging 1 3 s Start Pump Calibr Set Adjust Temp _ 683 pulses m Change Press Offset i Set show Feme Pump Calibrated OK 1005 mV on Press OK to continue Set Delay UV to Frac I 650 ul Set Flow Rate Enter Collected Volume Set Mixer Chamber Vol 3 0 ml min 3 0 0 00 ml 2 0 ml Set zero pressure Calibrating Offset to calib Press OK Done Press OK Ool 180 KTAprime plus User Manual 11 0026 44 Edition AA Reference information 11 3 Technical specifications Relevant system and component specifications are listed below 11 3 1 Operating data Pump Flow rate range 0 1 50 ml min in steps of 0 1 ml min Operating pressure range 0 1 0 MPa 10 bar 145 psi Pressure pulsation lt 10 of mean value in entire range PH stability range 1 14 spec valid between pH 2 12 Viscosity lt 10 ml min gt 10 ml min Max 10 cP Max 5 cP Flow rate reproducibility rsd lt 1 during one day pressure gt 0 1 MPa flow rate gt 0 4 ml min Flow rate accuracy 5
89. ped Volume 194529452ml When required contact GE Healthcare for replacement AkTAprime plus User Manual 11 0026 44 Edition AA 125 9 Maintenance Check Tube Shifts 17564 Check Valve Shifts BV 17564 IV 28143 126 9 7 Checking the fraction collector 9 7 1 Checking tube shifts 1 Select menu Check and press OK 2 Select menu Check Tube Shifts When required contact GE Healthcare for replacement 9 8 Checking the rotary valves 1 Select menu Check and press OK 2 Select menu Check Valve Shifts The value after BV shows the number of buffer valve shifts The value after IV the number of injection valve shifts One shift means the shifting between two adjacent positions The lifetime of the valves is gt 50 000 shifts When necessary replace the plates according to section 9 13 Replacing plates in the rotary valves or contact GE Healthcare for sealing replacement 9 9 Cleaning the UV flow cell in place WARNING NaOH is injurious to health Avoid spillage Pump a cleaning or sanitizing agent through the flow cell The standard recommendation is to pump 1 M NaOH for 30 minutes and then wash out immediately with buffer 9 10 Cleaning the UV flow cell off line A clean flow cell is essential for correct operation of the UV monitor CAUTION Do not allow solutions that contain dissolved salts proteins or other solid solutes to dry out in the flow cell Do not allow partic
90. piece of i d 0 75 mm PEEK capillary before cleaning the system flow path If not make sure that the column withstands the expected flow and pressures For column cleaning procedures and storage instructions please refer to the column instructions 9 3 1 Betweens runs Buffers not containing any salt can be left in the system for a short time after a run even overnight not in the pH electrode see instructions below If a buffer containing salt has been used the flow path should be flushed with deionized water This is especially important if an organic solvent will be used in the next run To flush the flow path 1 Filla syringe with five times the sample loop volume of deionized water 2 Rinse the sample loop by injecting the water through the fill port on the injection valve 3 Put all used inlet tubings in water 4 Inthe Templates menu select Application Template and then System Wash Method 5 Select the used inlet ports Inlets A1 and B will always be washed 6 Press OK to start the method The system flow path is automatically flushed AKTAprime plus User Manual 11 0026 44 Edition AA Maintenance 9 9 3 2 Weekend and long term storage CAUTION Never leave the pH electrode in the electrode holder for any period of time when the system is not used since this may cause the glass membrane of the electrode to dry out Store the pH electrode fitted in the end cover filled with a 1 1 mixture of pH 4 buffer and 2 M
91. plication template Method template Run Stored Method see page 177 Manual Run see page 178 Program Method see page 179 Copy Method see page 177 Set Parameters see page 180 Check see page 178 Gel filtration Sample Appl Volume 0 0 ml 0 0 Buffer Exchange lon Exchange Gradient elution HIC Gradient elution Affinity Step Gradient 176 Desalting i HiPrep De na IMAC Purification IgM Purification a i gai divi P ifi d Mab Purification Affinity Purification Tag Purification System Wash Method Select Buffer V Pos B A 77 OK Press OK to Start RUN Sample inject by To RUN data displays Injv Pump and Print out menus see page 179 Set Pressure Limit 1 00 MPa 1 00 Set Flow Rate Method Ready 1 0 ml min 1 0 Press OK Set Fraction Size 0 0 ml 0 0 Save Method Set Equilibr Volume yes yes no 0 0 ml 0 0 Set Sample Inj Vol Free Methods 25 0 0 ml 0 0 Sel Method used 09 Set Wash 1 Volume Clear Method 09 1 0 0 ml 0 0 yes ves n Set Elution Volume 0 0 mI 0 0 Set Wash 2 Volume Press OK to 1 0 0 ml 0
92. pump running while calibrating Switch to the second standard buffer solution and repeat the procedure AkTAprime plus User Manual 11 0026 44 Edition AA 141 9 Maintenance aR Set up pH temperature compensation The relationship between pH and the output signal from the pH electrode is temperature dependent For accurate measurements during temperature changes the pH measurement can be temperature compensated In normal applications when the temperatures of the buffers and calibration buffers are identical temperature compensation is not necessary When using temperature compensation it is important that the temperature of he pH electrode is the same as that of the conductivity flow cell since that is where the temperature is measured Set Parameters 1 From the main menu select sub menu Set Parameters and press OK Setup and 2 Select sub menu Setup and calibration Press OK calibration Setup pH 3 Select sub menu Setup pH and press OK Set pH Temp Comp 4 Select sub menu Set pH Temp Comp The current setting for showing pH is off displayed If on is shown Tc is displayed in the running display If off is shown default Tc is not displayed Press OK to change the setting Set pH Temp Comp 5 Select the desired setting and press OK off on off Ea 142 AkTAprime plus User Manual 11 0026 44 Edition AA Troubleshooting 10 Troubleshooting 10 1 Faults and actions This section
93. required see section 9 18 Compare the response of the pH electrode with that of another pH electrode If the response differs greatly the electrode may require cleaning or replacement 10 There may be interference from static fields Connect the pH flow cell to the rear of the system using a standard laboratory 4 mm banana plug cable 11 Check that the pH electrode has been calibrated at the correct temperature 12 Inorganic solvents such as ethanol methanol and acetonitrile stable pH measurements are not possible since dehydration of the membrane will occur We recommend that the pH electrode is not used in applications using organic solvents 13 Clogged liquid junction Refer to section 9 18 No response to pH changes 1 Check that the electrode cable is connected properly to rear of the system 2 The electrode membrane may be cracked If so replace the electrode Small response to pH changes 1 Clean the pH electrode according to section 9 18 and recalibrate 2 If the problem persists replace the pH electrode AkTAprime plus User Manual 11 0026 44 Edition AA 147 Troubleshooting el Fault Action Slow pH response or calibration 1 Check the electrode glass membrane If it is contaminated impossible clean the electrode according to the instructions in section 9 18 2 Ifthe membrane has dried out the electrode may be restored by soaking
94. ress the set key 13 i e set start position 7 Set the zero keys to the up position 8 Select a suitable chart speed 2 20 mm min according to the length of the purification 9 Select 1 V with the range V mV selector 1 10 The recorder is made ready to use by setting the rec key to on The chart paper starts rolling when the run starts sl 114 KTAprime plus User Manual 11 0026 44 Edition AA Set Parameters Set Analogue Out Set Rec Out 1 UV Set Rec Out 1 UV UV pH Cond B Tmp Pr Set UV Analogue Out 0 005AUFS 10 Set UV Zero Level 10 0 10 0 Set UV Range 0 005AUFS 0 0002 i i KTAprime plus User Manual 11 0026 44 Edition AA Installing and modifying components 8 8 6 5 Setting analog outputs The Set Analogue Out menu is used to select which measurement parameter that should be associated to each channel The menu is also used for setting measurement parameters zero and full range values for UV Cond pH and Press on the analogue output channels 1 Select the Set Parameters menu in the main menu and press OK 2 Select the Setup Analogue Out menu and press OK to enter the settings menu Setting parameters for the channels UV pH conductivity concentration of buffer B temperature and pressure are measurement parameters that can be associated to the analogue output channels 1 When entering the Setup Analogue Out menu the
95. t NC of the flow diversion valve Tube Sensor 108 AkTAprime plus User Manual 11 0026 44 Edition AA Installing and modifying components 8 8 5 pH flow cell and electrode optional 8 5 1 Mounting the flow cell holder In the AKTAprime plus system the pH electrode is optional 1 Hook the flow cell holder on the right hand side of the housing Secure it with the slide clamp If the flow cell holder is not used the flow cell must still be installed at an angle of 30 from the vertical with the outlet placed higher than the inlet to prevent air bubbles S being trapped in the cell ra The flow direction is marked on the flow cell Flow direction 2 Connect the tubing with finger tight connectors 8 5 2 Inserting the pH electrode Note Handle the pH electrode with care CAUTION The tip of the pH electrode consists of a thin glass membrane Protect it from breakage contamination and drying out or the electrode will be destroyed Always store the electrode with the end cover filled with a 1 1 mixture of pH 4 buffer and 2 M KNO3 Do NOT store in water only 1 Unpack the pH electrode Ensure that it is not broken or dry 2 Before using the electrode remove the electrode end cover and immerse the glass bulb in buffer for 30 minutes 3 Carefully insert the electrode in the flow cell Tighten the nut by hand To system rear panel Note Ifthe flow cell is full of liquid it is not possible to
96. terature Protein Purification Handbook Affinity Chromatography Principles and Methods Gel Filtration Principles and Methods Hydrophobic Interaction Chromatography Principles and Methods lon Exchange Chromatgraphy amp Chromatofocusing Principles and Methods Antibody Purification Handbook AKTAprime plus User Manual 11 0026 44 Edition AA 11 0026 44 11 0027 48 18 1132 29 18 1022 29 18 1022 18 18 1020 90 11 0004 21 18 1037 46 Index Index A Accessories and consumables Application templates B Breakpoints editing inserting deleting Buffer preparation Buffer valve replacing parts troubleshooting Cc COlIDatIONS a a a acd a carn ab hd ih ahs nda RE S Changing a parameter value Check menus Check valves Checking communication Chemical resistance Cleaning the flow path Cleaning the system Collecting fractions Column pressure Columns and tubing Communication cable Complete filling Components 05g 01110 AE E EEOAE A ENEE EEE AE ENET 156 location Conductivity flow cell installing Connecting columns Control keys Copying a method D Data presentation PrimeView Recorder REC 112 During a run changing parameters completing a run interrupting a run viewing progress E Editing a stored method Electrical connections Error messages AkTAprime plus User Manual 11 0026 44 Ed
97. the inlet filters visually and replace them if necessary Every month Monitor Check the monitor according to section 9 5 Flow restrictor Check that the flow restrictor generates the following back pressure 0 2 0 05 MPa Check the back pressure as follows 1 Disconnect the flow restrictor 2 Connect a capillary to port 1 of the injection valve 3 Run the pump manually at 10 ml min with water Note the backpressure on the display 4 Connect the flow restrictor to the open end of the capillary 5 Runthe pump at 10 ml min with water Note the back pressure on the display 6 Calculate the backpressure generated by the flow restrictor Replace it if it is not within limit 120 AKTAprime plus User Manual 11 0026 44 Edition AA Maintenance 9 Interval Action Every 6 months or more often if required Monitor e Clean the UV flow cell according to section 9 9 or 9 10 e Change the pH electrode Refer to section 8 5 Fraction collector e Check the drive sleeve on the tube rack Replace if worn e Check the number of tube shifts according to section 9 7 Superloop e Check that the top bottom and moveable seal O rings are in good condition Replace if necessary e Check that the bottom end pieces are clean and undamaged Mixer e Check that the mixer chamber is clean and without damage e Check the tubing connectors Replace if required Refer to section 9 17 Yearly
98. to change the setting 5 Change the setting as desired and press OK AkTAprime plus User Manual 11 0026 44 Edition AA 169 Reference information Set Parameters Setup and calibration Setup UV Set Averaging 1 3 s Set Averaging 1 3 s 0 64 Se 170 Set up UV averaging filter constant To filter the noise in the UV signal a moving average filter is used The averaging time is the time interval used for calculating the moving average of the absorbance signal A long averaging time will smooth out noise efficiently but it will also distort the peaks Peaks narrower than the minimum peak width value according to the table below may be distorted Because of this the averaging time should be as short as possible On delivery the averaging time is set to 1 3 s 1 From the main menu select menu Set Parameters and press OK 2 Select sub menu Setup and calibration and press OK 3 Select sub menu Setup UV and press OK 4 Select sub menu Set Averaging The current set averaging time is shown Press OK to change the setting 5 Set the desired value and press OK Values allowed are 0 02 0 04 0 08 0 16 0 32 0 64 1 3 2 6 5 and 10s Averaging time s Corresponding time Min peak width constant s at half height s approximately 10 0 5 50 5 1 2 32 2 6 1 16 1 3 0 5 8 0 0 64 0 2 3 2 0 32 0 1 1 6 0 16 0 05 0 8 0 08 0 03 0 5 0 04
99. tors 8 4 Fraction collector 8 4 1 Assembling the tube rack There are three types of tube racks Rack Max no tubes Tube diam Tube length 12mm 175 12mm 50 180 mm 18 mm 95 10 18 mm 50 180 mm AkTAprime plus is delivered with the 18 mm rack mounted Each rack is made up of a combination of a bowl tube supports tube guide and tube holder The 12 mm rack is available as accessory Also available for use with the 12 mm tube rack is a double ended Eppendorf tube holder 18 8522 01 One end holds 1 5 ml Eppendorf tubes the other holds 0 5 ml tubes AKTAprime plus User Manual 11 0026 44 Edition AA Installing and modifying components ey Single cutout L shaped cutout f Note that the tube guide has both single Tube Guide and L shaped cutouts while the tube Tube Support BON holder only has single cutouts When assembling a rack different cutouts are used for the various items depending on the length of the tubes The cutouts are summarized in the tables below Tube racks 12 and 18 mm Item 50 85 mm tubes 85 100 mm tubes Tube support L shaped cutout Not required Tube guide Single cutout L shaped cutout Tube holder Single cutout Single cutout 1 Insert the tube support if required in the bowl The circular marks on the support should face down 2 Insert the tube guide with the tube numbers facing upwards The guide should rest about 1 cm above the support N 3 Insert the tube holder checking that positi
100. ty flow cell Conductivity flow cell pH electrode pH electrode round tip incl flow cell and holder pH electrode round tip pH flow cell round tip incl dummy electrode Dummy electrode round tip 1 18 1111 05 1 C 18 1134 84 1 C 18 1111 26 1 A 18 1112 92 1 A 18 1111 03 KTAprime plus User Manual 11 0026 44 Edition AA Reference information SS Item Quant A C Code no pack Injection valve Repair kit INV 907 1 C 18 1109 05 Injection fill port 0 7 mm 1 C 18 1127 66 Buffer valve Repair kit IV 908 1 C 18 1109 07 Mixer Mixing chambers 0 6 ml 1 A 18 1118 90 2ml 1 A 18 1118 91 5ml 1 A 18 1118 92 12 ml 1 A 18 1118 93 Fraction collector Tube racks complete with bowl tube support holder and guide 12 mm 1 A 19 8684 03 18mm A 18 3050 03 Tube support 1 A 18 3054 02 Tube holder and guide 12 mm 1 A 19 7242 02 18 mm 1 A 19 8689 02 Eppendorf tube holder for 12 mm rack 100 A 18 8522 01 Flow diversion valve FV 903 1 A 18 1114 50 incl mounting bracket Tubing holder A 18 6464 01 Drive sleeve C 19 6067 02 Sample loops 10 ul 1 C 18 1120 39 100 ul 1 C 18 1113 98 500 ul 1 C 18 1113 99 1ml 1 C 18 1114 01 2ml 4 C 18 1114 02 5ml 1 G 18 1140 53 Superloop 10 ml 50 ml Superloop 10 ml complete 1 A 18 1113 81 Superloop 50 ml complete 1 A 18 1113 82 Inner end piece 1 A 19 7846 01 Outer end piece 1 A 19 5167 01 O ring inner end piece 5 G 19 7595 01 O ring movable seal 2 G 18 1104 97 n
101. uS cm to 999 9 mS cm Reproducibility short term Max 1 or 5 uS cm whichever is greater long term Max 3 or 15 uS cm whichever is greater Noise Max 0 5 of full scale calibrated range Response time lt 3 s 0 95 of step Temperature sensor accuracy 2 0 C drift 0 5 C per 10 h Flow rate sensitivity 1 within 0 100 ml min Conductivity flow cell Flow rate 0 100 ml min Max pressure 5 MPa 50 bar 725 psi Max backpressure 0 01 MPa at 100 ml min Fraction collection Tube capacity 95 in tube rack 18 mm 175 in tube rack 12 mm optional A 182 KTAprime plus User Manual 11 0026 44 Edition AA Reference information pH measurement pH range 0 to 14 spec valid between 2 and 12 Accuracy temperature compensated 0 1 pH within 4 to 40 C not compensated 0 2 pH within 15 to 25 C 0 5 pH within 4 to 15 C and 25 to 40 C Response time lt 10 s 0 95 of step Long term stability Dev max 0 1 pH per 10 h at constant conditions 4 40 C Flow rate sensitivity Dev max 0 1 pH units pH cell Flow rate 0 1 100 ml min Max pressure 0 5 MPa 5 bar 72 psi Max backpressure 0 02 MPa at 100 ml min KTAprime plus User Manual 11 0026 44 Edition AA 183 Reference information 11 3 2 Physical data Control Via membrane keyboard and display 2x20 characters Degree of protection housing IP 20 flow cells IP 44 Power requirement 100 120 220 24
102. valves in the system one is used for gradient formation and the other one for flow diversion during fractionation Pump The pump is equipped with a single input and double outputs only one is used It contains three internal plunger pumps which in combination with check valves create a smooth flow from the pump The pump delivers flows up to 50 ml min and pressures up to 1 0 MPa In the connection part the inlet and outlet flow paths are split into three separate flow paths one to from each plunger pump chamber All six flow paths are equipped with non return check valves Outlet port Check valve x6 Plunger pump Connection chamber x3 part Leakage hole Inlet port _ The plungers pump the liquid through the chambers while the check valves prevent the liquid from flowing backwards The pump phases of three plunger pumps are displaced by 120 which results in a sequential motion of the plunger pumps and thereby a smooth liquid delivery Leakage between the connection part and the pump chambers is prevented by O rings Any leakage behind the plungers is diverted through the drainage hole in the front of the connection part The check valves are also equipped with O rings The pump is equipped with automatic speed control to reduce pulsation and with pressure compensation AkTAprime plus User Manual 11 0026 44 Edition AA 157 Reference information Pressure

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