1 - GE Healthcare Life Sciences
Contents
1. Time eA13 Man Peak Fractionation Peak_Fractionation enables only peaks to be collected The UV mon itor is used to determine when there is a peak to collect Peak size deter mines max volume to be collected in each tube To collect the peak in lone tube set PeakSize gt peak volume If Peak FracParameters are not set in the method default values in sys tem settings will be used Tube change is delayed with respect to the delay volume To stop Peak Fractionation use the instruction Peak FracStop Parameters Rack The rack used Frac order The fractionation order kan be selected as Serpentine row Row by row Serpentine column or Column by column Tube type The tube type to frac tionate in PeakSize The size of the peak frac tions Start at Where to start the fraction ation if a specific position is desired use Tube number Base The peak fractionation can be performed in time or volume base Rack Frac order Serpentine row Row by row Serpentine column Column by col umn Tube type PeakSize 0 000 99999 999 base StartAt First tube Next tube Next line Next group Tube number Base Volume Time Ignore_LastTube Allows for fractionation after the last tube position has been reached A 2 4 Alarms amp Monitors This group contains the same instructions as the Alarms amp Mon group in method instructions Section A 1 4 um 18 1138 53A2. UVlampOFF Sets the UV lamp OFE This command only influences the specific run A 1 5 Watch Instruction Description Parameters Hold Until Puts the method on Hold Signal until a specific condition UV signal test or value of Cond the instruction is met or ppH the time out limit for the Pressure instruction is reached Test see below Value TimeOut 0 0 999 9 INFINITE um 18 1138 53AB e A10 Watch_UV1 Sets a watch on the signal Test Watch_Cond UV1 for UV to perform see below Watch_Conc la set action if a given test Value Watch_pH condition is fulfilled The Action Watch_Pressure watch is cancelled auto see below Watch_SamplePressure matically when the condi Value Watch_SampleFlow tion has been fulfilled once Watch_Off Cancels a watch on a Signal specified monitor Pressure Conc UV pH Cond Test Action Greater_than Block names Less_than Ready Slope_greater_thanLes Pause s_than_or_ Hold valley End Method Peak max End Block Stable baseline Continue A 1 6 Other Instruction Description Parameters Base Defines the base for a block for calcu Column Any lating breakpoints Each block must column name have a defined base Base Time Volume ml CV Same AsMain Block Calls a block unconditionally Block name Continue Resumes execution of a paused or held method This instruction has the same effect as clic
3. ductivity is displayed as a percentage lof bufferB in run data Cond and as la curve Cond CondValue 0 999 9 mS cm um 18 1138 53AB A18 CondTempComp Relates conductivity to concentration and or compares conductivity values The compensation consists of a com pensation factor together with a refer ence temperature CondRef Temp All conductivity values will then automat ically be converted to the set reference temperature The factor is expressed in percentage increase of conductivity per PC increase in temperature If the CompFactor is unknown a general lapproximate value of 2 can be set for many common salt buffers 0 off CompFactor 0 9 9 CondRefTemp Selects the reference temperature to which the measured conductivity val ues will be converted CondRefTemp is active when a factor of CondTemp Comp is selected RefTemp 0 99 9 C pHTempComp Sets the temperature compensation on lor off For more accurate measure ments during temperature changes the pH measurement can be temperature compensated When using pHTemp Comp it is important that the tempera ture of the pH electrode is the same as that of the conductivity flow cell since that is where the temperature is meas lured pHTempComp ON OFF WatchPar_UV1 WatchPar_Cond WatchPar_Conc WatchPar_pH WatchPar_Pressure WatchPar_Sample Pressure WatchPar_Sample Flow The Delta Peak setting affects
4. 11 A 2 Manual control 000000 cence eee es A 13 AAI PUMPS dorada a ee we aL are te Ree eee aa SE A 13 Aziz FlOWPatrs gas asia a Siete Aine Deven a eae A eae A 13 ADS FAC O da aan A E A 13 A 2 4 Alarms amp Monitors 0 00 00000000000 eee ee A 14 A2 5 OMCs 8 oer a at Be ths A 15 A 3 System settings instructions A 15 ASol Alarms rs 22 s a aula gem toas ahead A 15 A32 Special Sisi ay adn hae Ata eee ee Pees A 17 A33 MONS Avie act a ta Gee iaa A 18 ASA AAA A A A 20 B Technical Specifications B 1 System Requirements o ooococoococn eens B 1 epl e p2 B 1 1 Hardware requirements 0 0 0 eee ee B 1 B 1 2 Software requirements 0 0 0 0000 es B 1 B 1 3 Network requirements 0 0 00 cc ees B 1 B 2 Control Capacity 20 20 e eee eee B 2 B 2 1 Stand alone installatiONS ooooo o o oooooo oo B 2 B 2 2 Network installatiONS ooooo oo ooooooooooo B 2 C Installation of the PCI expansion card C 1 INDEX 2 ua ic i 1 Frac 950 Setup and Documentation 1 1 Introduction UNICORNTM is a control system developed and marketed by Amersham Biosciences AB for real time control of chromatographic separation systems from a personal computer The package operates together with systems in the AKTA design platform referred to as AKTAdesign systems BioPilot System and BioProcess System from Amersham Biosciences Standard configurations are pr
5. by its edges and as far as possible avoid touching the electronic components Dip switch settings Selected bed pac Control Unit expansion card fz im 1 2 4 onjm 1 E 1 2 2 ON Cm 1 Caj2 3 onNjm 1 1 Caj2 4 ON CM 1 Figure C 1 The PCI expansion card and dipswitch settings for the CU ecl um 18 1138 53AB e C2 Set the control unit CU setting by using a dip switch To check the CU setting compare the expansion card with the settings shown in Fig C 1 Record the number of the set CU During installation of UNICORN software the Setup program will request these card settings Enter the correct value for the selected CU so that UNICORN can find the systems Close the cover of the PC Connect the expansion card to the liquid handling module CU connector using the communication cable provided Additional liquid handling modules up to a total of four may be connected INDEX A alarms and monitors instructions A 9 alarms and monitors manual control instructions A 14 alarms system settings instructions A 15 c chromatography systems used with UNICORN 3 curve system settings instructions A 20 D documentation for manual runs 5 printing 7 viewing the Frac 950 tab 7 F flowpath instructions A 4 manual instructions A 13 frac manual instructions A 13 frac instructions A 6 Frac 950 summary of functional features 3 tab within Documentati
6. chromatography systems This is achieved using the four possible System control windows in UNICORN available on each workstation Each chromatography system may be controlled by only one active System Control window and be viewed by seven different other System control windows in UNICORN C Installation of the PCI expansion card Information for service personnel In most cases UNICORN is delivered pre installed by qualified service personnel The installation procedure described below should not be attempted by unqualified personnel The PCI expansion card replaces the ISA cards for the CU 900 control board supplied in earlier computer models The upgrade is needed only for stand alone installations i e systems not connected to a network and for local computers in network installations Note This expansion card cannot be used with other CU units Note Pentium 2 computers will require a BIOS update available from www compaq com support files desktops us index html 1 Turn off the power to the PC and unplug the main power cables 2 Open the PC cover see manufacturer s instructions 3 Locate an empty PCI expansion slot Caution Electrostatic discharges can permanently damage electronic components on the card Discharge any electricity from your body by touching an earthed metal surface e g a water tap or radiator before handling the card 4 Remove the expansion card from the anti static bag Handle the card
7. cuted and the run is continued min by pressing Continue the sample loading will be stopped and the run will con tinue PumpWashEx PumpWash is used to change InletA1 plorer eluents in the inlet tubings OFF pump and mixer THE 411 418 PUMPS MUST BE PURGED InletA2 PRIOR TO USING PUMP OFF WASH Flow is diverted to ON the waste position of the InletB1 injection valve Up to four OFF inlets can be washed simulta ON neously InletA1 InletA2 InletB2 InletB1 or Inlet B2 One ofthe OFF eight inlets of buffer valve A1 ON can be selected for InletA1 The system is set to Pause during PumpWash and the pressure limit is set to the value given in instruction Wash alarm pressure in Sys tem Settings After Pump Wash the injection valve turns back to its previous position and the flow and pressure lim its are set to their previous values Flow rate 25 ml min Wash volume 30 ml inlet PumpWashPurifier PumpWash instructions for PumpWashBasic AKTApurifier and AKTAbasic respectively um 18 1138 53AB e A2 SystemWash System Wash is used to change eluents in the system includ ing inlet tubings pumps mixer and flow cells THE PUMPS MUST BE PURGED PRIOR TO USING SYS TEMWASH Flow is diverted to the waste at the outlet valve The ColumnPosition is set to Position1Bypass during System Wash One inlet can be selected A11 A18 When run ning BufferPrep methods the System Wash instructio
8. only the Less than or valley and Peak max conditions when using the Watch instruction see UNICORN User Manual As a general guideline set the value to 2 3 times the noise level and 5 10 of the smallest expected peak height Delta_Base set ting affects only the Stable_baseline Condition For this condition to be activated the signal may not vary by more than the Delta Base value above lor below the time interval specified in the Stable_baseline condition in the watch instruction Delta_Peak Delta_Base eA19 A ASA Curves Instruction Description Parameters UV1 Set the specific signal curve on off for Store UV2 storing in the Result file The time ON OFF UV3 between samples determines the fre Time between Cond quency with which curve data is samples Cond recorded Conc pH Pressure Flow Temp Sample_Pressure Sample_Flow um 18 1138 53AB e A20 B Technical Specifications B 1 System Requirements B 1 1 Hardware requirements e Pentium 11 333 MHz or later minimum Pentium 90 MHz e 64 Mb RAM minimum 32 Mb for one system 128 Mb RAM minimum 64 Mb for two or more systems e 1 Gb of available hard disk space 150 Mb to run UNICORN NTFS file system e Colour monitor 1024x768 pixels minimum 800x600 small fonts 64k colours e 1ISA or 1 PCI slot per connected system e CD ROM drive e 1 44 Mb 3 5 diskette drive e Mouse option when industrially har
9. width of the narrowest peak to collect Keyboard Sets the mode of the user interface of Mode the instruments Open KeyLocked KeyAndDial Locked e A17 A 3 3 Monitors ductivity signal The relative conduc tivity is displayed as a percentage of bufferB in run data Cond and as a curve Cond Instruction Description Parameters Wavelength Sets the wavelength and number of UV1 wavelengths to monitor The range is 190 700 nm 190 700 nm for UV1 UV2 and UV3 UV2 0 OFF 0 OFF for UV2 and UV3 Wave 0 700 nm lengths lt 190 nm are automatically set UV3 0 OFF to O off 0 700 nm AveragingTimeUV Filters the noise in the UV signal Aver AvTimeUV aging time is the time interval used for 0 01 calculating the moving average of the 0 02 absorbance signal A long averaging 0 04 time will smooth out noise efficiently 0 08 but will also distort the peaks Choose 0 16 an averaging time of the greatest value 0 32 that can be set which is less than 1 0 64 11 60 s min typical peak width ml 1 28 flow rate ml min The averaging 2 56 time should not be less than 1 28s 5 12 when more than one wavelength is monitored A column specific averag ing time is set automatically when a column and a template are selected SetCondScale 0 Pets the 0 scale of the relative con CondValue 0 999 9 mS cm SetCondScale 100 Sets the 100 scale of the relative conductivity signal The relative con
10. 12mmTubes Microplates 3OmmTubes Figure 1 2 The Rack drop down menu within the Frac 950 tab um 18 1138 53AB e p4 A 1 2 2 Fraction order Next use the Fraction order radio buttons to choose the appropriate order for fractionation Figure 1 3 The Fraction order radio buttons 1 3 System control In System control for manual runs the Frac 950 tab cannot be used in Documentation Instead use the manual fractionation instructions starting with Man see also Appendix A Manual control 1 3 1 Last tube The last tube position can only be selected from within Start Protocol before starting a run Click the Define box within the Last tube panel and then move the cursor over the appropriate tube circle within the tube matrix and click again ep5 Frac 950 x in 00090 TY TITO pi R 900000000000 e Row by 0806800060 400 z 8606000000000 r Segpenine f f j 000000000000 E 000000000000 a 1333331337 200000000000 Set default Tisya 000000000000 r 0000 DEM Hep lt Back Cancel Figure 1 4 The Frac 950 tab illustrating the Last tube function If the fractionation process proceeds beyond this position during a run an alarm will be executed The lower right box within the Last tube panel shows the currently selected last tube If a new last tube has been chosen but the user wishes to return to the default last tube position simply click on the Set default button This will
11. Alarm High Alarm will set the system in Pause 0 1 0 MPa Low Alarm 0 1 0 MPa um 18 1138 53AB A16 A 32 Specials Instruction Description Parameters FracParameters Parameter settings for fraction collec DelayVol tor Delay volume is defined as the vol 0 10 ml lume from UV flow cell to the outlet of TubeChange the fraction collector Collection of Accumulator flow during tube change can be han DropSync dled in different ways Accumulator Tube The flow will be collected in the accu mulator during tube change and then be delivered in the next tube DropSync Tube change synchronised to drop release Should only be used at flow rates generating drops below 5 ml min if capillary 0 5 mm id is used and below 3 ml min for 0 25mm id tubing Tube No synchronisation of collection Note The delay volume must be changed when the parameter TubeChange is altered to or from Accumulator see the manual for the correct delay volume Peak_FracPara UV1 parameter settings for StartSlope meters Peak Fractionation To obtain suitable 0 01 10000 values perform a test run and measure mAu min the slope at the beginning and end of EndSlope the smallest and flattest peak you want 0 01 10000 to collect using Operations Differen ImAU min tiate in Evaluation Time min as base MinWidth must be used Minwidth prevents col 0 01 1500 min lection of noise spikes Enter a value of about half the
12. B A14 A 2 5 Other Instruction Description Parameters Block Calls a block Only available when a Block method is running Block name Next breakpoint Jumps to the next breakpoint in the current method only relevant when a method is running BufferPrepRecipe Pelects a recipe for running BufferPrep Recipe Name manually The recipe must be selected las the first instruction for the manual run Set Mark This inserts a specified text into the chromatogram End_Timer This is used to set the time out limit in Accumulated time or volume for the End instruc time or vol tion ume 0 9999 min or 0 9999 l or disabled Record_on Begins recording a run that has been started manually A result file will be generated A 3 System settings instructions A 3 1 Alarms Instruction Description Parameters Alarm UV1 Sets the alarm limits for the signal Mode Alarm_Cond An alarm will set the system in Disabled Alarm pressure Pause Enabled Alarm_pH HighAlarm LowAlarm e A15 Wash Alarm pressure Sets the alarm limits for pressure Mode when instruction PumpWash or Disabled System Wash are used This pres Enabled sure limit only applies when these High Alarm instructions are executed 0 10 MPa Low Alarm 0 5 MPa Alarm SamplePressure Sets the alarm limits for sample Mode pressure An alarm Disabled SamplePressureHighAlarm Enabled SamplePressureLow
13. Q rona chromatography software UNICORN version 3 20 Frac 950 module for UNICORN applications Amersham um 18 1138 53AB 2000 07 e Biosciences to order Asia Pacific Tel 852 2811 8693 Fax 852 2811 5251 Australasia Tel 61 29894 5152 Fax 61 29899 7511 _ Austria Tel 01576 0616 20 Fax 01576 0616 27 Belgium Tel 0800 73 888 Fax 03 272 1637 Canada Tel 1 800 4635800 Fax 1 800 567 1008 Central East South East Europe Tel 43 1 982 3826 Fax 43 1 985 8327 Denmark Tel 45 16 2400 Fax 4516 2424 Finland Tel 095123940 Fax 095121710 France Tel 0169356700 Fax 0169 41 9677 Germany Tel 0761 4903 401 Fax 0761 4903 405 Italy Tel 02 27322 1 Fax 02 27302 212 Japan Tel 813 5331 9336 Fax 81 3 5331 9370 LatinAmerica Tel 55 11 3667 5700 Fax 55 11 3667 8799 Middle East and Africa Tel 30 1 96 00 687 Fax 30 1 96 00 693 Netherlands Tel 0165 580 410 Fax 0165 580 401 Norway Tel 2318 5800 Fax 2318 6800 Portugal Tel 01 417 7035 Fax 01 417 3184 Russian Federation Tel 7 095 232 0250 956 1137 Fax 7 095 230 6377 South East Asia Tel 60 3 724 2080 Fax 60 3 724 2090 Spain Tel 93 594 49 50 Fax 93 594 49 55 Sweden Tel 018 6121900 Fax 0186121910 Switzerland Tel 01 802 81 50 Fax 01 802 81 51 UK Tel 0800 616 928 Fax 0800 616 927 USA Tel 1 800 526 3593 Fax 1 800 329 3593 BioPilot BioProcess UNICORN and AKTA are trademarks of Amersham Biosciences Limited or its subsidiaries Amersham is a trademark of Nycomed Amersha
14. automatically set the last tube to its default position Note When using different sized tubes racks 18mm tubes or Microplates El um 18 1138 53AB e p6 the last tube can be set for both tube sizes Use the Tube type drop down menu to choose the desired tube size and then follow the procedures outline above to select the last tube Note that the total number of tubes sampled may differ according to which Fraction order has been chosen The Number of tubes equation in the bottom left corner of the Frac 950 tab shows the current number of available tubes chosen for fractionation followed by the total possible number in parentheses For example in the figure above 120 of the total 120 small tubes and all 8 of the large tubes will be available for fractionation 1 4 Evaluation 1 4 1 Documentation The Frac 950 tab can be displayed within the Evaluation module by choosing View Documentation or pressing the Documentation icon on the toolbar and pressing the Frac 950 tab You can print out the selected settings by pressing the Print button or return to the main Evaluation window by pressing OK or Cancel Documentation x Variables Scouting Text Method Questions Notes BufferPrep Columns Reference Curves Evaluation Procedures Method Information ResultName StatProtocol Settings Calibration Logbook Evaluation Log Frac 950 Fraction order Help Print DK Figure 1 5 T
15. ave This directs you to click on the File menu and select the command Save As A dialogue called Save As is displayed in which you must locate the destination folder for the saved file and give the file a name You then click on the button called Save in the dialogue to execute the save command ep3 1 2 Method editor Run Setup User defined options for Frac 950 are selected on the Frac 950 tab in Run Setup in the Method editor To reach Method editor from the Main menu create a new method as described in the UNICORN 3 10 manual To access the Frac 950 tab in Run setup either click on the Run setup icon on the toolbar or select View Run Setup from the menu The Frac 950 tab allows the user to choose rack type the fractionation order and the last tube position Note The Frac 950 tab is only available if your strategy supports Frac 950 Variables Scouting Questions Notes Gradient BufferPrep Columns Reference Curves Evaluation Procedures Method Information Result Name Start Protocol Frac 350 Rack 18 amp 30 mmT ubes r Fraction order Setpentine 22222277 o III o 0006000000000 000000000000 cs 1 000000000000 0006000000000 O 4060000000009 000000000000 000000000000 0000 Help Figure 1 1 The Frac 950 tab 1 2 1 Rack type Begin the fractionation set up by choosing the appropriate rack type from the Rack drop down menu Rack 18mmTubes 18mmT ubes
16. dened screens are required B 1 2 Software requirements Microsoft Windows NT Workstation 4 0 with Service Pack 5 or later B 1 3 Network requirements These are the recommended network requirements for running UNICORN in a network installation e Supported Network cards 3COM Etherlink III Compaq Netelligent 10 100 TX Embedded UTP Controller Compaq Integrated NetFlex 3 Controller AMD PCNET PCI Ethernet Adapter Integrated e Novell NetWare version 4 50 189 or later or Microsoft Windows NT Server 4 0 The UNICORN software works on earlier versions as well even though some versions of the Novell Netware driver have known problems e A valid network connection e B1 B2 Control Capacity B 2 1 Stand alone installations Simultaneous control of up to four chromatography systems Each module is separately configured in a system strategy supplied by Amersham Biosciences B 2 2 Network installations A few basic facts about a network installation um 18 1138 53AB e B2 Chromatography systems must be locally linked to a workstation and then the workstation is linked to the network i e the chromatography systems are not directly linked up to the network Each local workstation can be connected to four separate chromatography systems A network can support up to 90 chromatography systems which are connected locally to the workstations in the network A workstation can locally or remotely actively control up to four
17. e length is monitored A column specific averaging time is auto matically set when a column and template are selected Alarm UV1 Sets the alarm limits for the sig Mode Alarm Cond nal An alarm will set the system Disabled Alarm_pH in Pause Enabled Alarm Pressure HighAlarm Alarm SamplePressure LowAlarm SetCondScale 0 Sets the 0 scale of the relative CondValue conductivity signal The relative 0 999 9 conductivity is displayed as a per mS cm centage of bufferB in run data Cond and as a curve CondY SetCondScale 100 Sets the 100 scale of the rela CondValue tive conductivity signal The rela 0 999 9 tive conductivity is displayed as a mS cm percentage of bufferB in run data Cond and as a curve CondY WatchPar_UV1 The Delta Peak setting affects Delta Peak WatchPar cond only the Less than or valley and Delta Base WatchPar_Conc Peak_max_conditions when WatchPar_pH using the Watch instruction see WatchPar_Pressure UNICORN User Manual chap WatchPar_SamplePressure ter 5 8 8 As a general guideline WatchPar_SampleFlow set the value to 2 3 times the noise level and 5 10 of the smallest expected peak height Delta_Base setting affects only the Stable_baseline condition For this condition to be acti vated the signal may not vary by more than the Delta Base value above or below beyond the time interval specified in the Stable baseline condition in the watch instruction
18. he Frac 950 tab in Documentation in the Evaluation module 1 4 2 Fraction text alignment In the Evaluation module click on the chromatogram with the right hand mouse button select Properties from the menu and then select the Curve Style and Colour tab This will allow you to select Vertical Horizontal or Fly Over orientation for the Fraction text mark ep um 18 1138 53AB e p8 Chromatogram layout 1 07 id2301 1_Pressure id2301 1_Flow 10 id2301 1_Temp idIINI 1 _Cesoticma Figure 1 6 The chromatogram layout dialogue with the Fraction text alignment box highlighted To change the fraction text alignment from the default option horizontal select the appropriate radio button and then click on the OK button PA id2301 1 id2301 1_UV1_280nm 142301 1_UV2_Onm id2301 1 UV3 Onm id2301 1 Cond id2301 1 Cond id2301 1_ id2301 1 Flow id2301 1 Temp id2301 1_Fractions id2301 1_Inject Vertical fraction marks Figure 1 7 A chromatogram window in the Evaluation module showing Vertical fraction text mark alignment A Strategy for AKTAexplorer 100 A dedicated strategy is available for each system in the AKTAdesign platform The instructions available differ slightly between strategies although the majority are common To find a description of any instruction in your particular strategy mark the instruction and click on lt F1 gt To obtai
19. he Outlet valve Feed switches the outlet valve one step forward from its actual posi tion The OutletValve gives fraction marks Position WasteF1 FracF2 F3 F8 Feed BufferValveA1 Selects the position for Buffer Valve A1 Position A11 corresponds to position 1 on the Buffer Valve A1 position A12 to position 2 etc The flow rate is reduced to O ml min when the valve is switched Position A11 A18 PumpAlnlet Sets the position of the pump A inlet The instruction is not allowed in a method when running BufferPrep gives an alarm in BufferPrep mode Position A1 A2 PumpBlInlet Sets the position of the pump B inlet The instruction is not allowed in a method when running BufferPrep gives an alarm in BufferPrep mode Position B1 B2 SampleValve Selects the position for the Sample Valve Position S1 S8 FlowDirection Sets the flow direction for the col umn The instruction is useful for example when cleaning columns The instruction switches the Flow Direc tion Valve to position 2 with parame ter UpFlow and to position 1 with parameter DownFlow The flow rate is reduced to O ml min when the valve switches Direction Downflow Upflow InjectionMark The instruction gives an injection mark The instruction is useful when the sample is introduced onto the col umn with the gradient pump A 1 3 Frac Instruction Descripti
20. king on the Cont button in System control End_Method Terminates method execution equiva lent to clicking on the End button in System control e All Evaluate Calls an evaluation procedure The procedure must be stored together with the current method Procedure name Hold Places the system in Hold state This instruction has the same effect as clicking on the Hold button in System control Loop Runs the instructions between Loop and Loop end for the specified number of times No of loops 1 9999 Loop end Specifies the end of the instructions contained within a loop Message Generates a user defined message which is recorded in the log book and may be displayed on the screen Message Mode Screen INoscreen INew_ chromato gram A new chromatogram is created within the result file and all data col lected after this instruction is stored within the newly created chromato gram until another New chromato gram instruction is issued Name Pause Places the system in the Pause state for the specified length of time Time 1 infinite 9999 9 in min utes Ready Indicates that the next step in a MethodQueue may start Set_Mark Inserts a specified text into the chromatogram End_Block Terminates a block and returns con trol to the point from which the block was called Inserts a comment in the me
21. m plc Microsoft and Windows NT are trademarks of the Microsoft Corporation Amersham Biosciences AB Bj rkgatan 30 SE 751 84 Uppsala Sweden Amersham Biosciences UK Limited Amersham Place Little Chalfont Buckinghamshire HP7 9NA England Amersham Biosciences Inc 800 Centennial Avenue PO Box 1327 Piscataway NJ 08855 USA Amersham Biosciences Europe GmbH Munzinger Strasse 9 D 79111 Freiburg Germany All goods and services are sold subject to the terms and conditions of sale of the company within the Amersham Biosciences group that supplies them A copy of these terms and conditions is available on request O Amersham Biosciences AB 2000 All rights reserved CONTENTS Frac 950 Setup and Documentation 1 14 IntroduCii nh 0 00 a cd 3 1 2 Method editor Run Setup ooocoooomcocommmomo 4 FZI AAA A E 4 1252 la 5 1 3 System control oocoococco eee 5 RES A E AA A RR E RE RN 5 1 4 Evaluation ocana eee Satay A ES a e 6 1 4 1 Documentation a ss 3 4 bd dd it ld 6 1 4 2 Fraction text alignment 0 0 0 0 eee 7 A Strategy for AKTAexplorer 100 A 1 Method instructions 0000 cee e eee A 1 ALI PUMP ae crt ease as avr nape Da sewed exe de A 1 ATEZ FlOWPatl ire ses o dA CREU etna Bint acerca eee Ve A 4 ARES FRAC tl cote ffs cies ange punish ed pda Bes A 6 A 1 4 Alarms amp Monitors 2 0 00 0000000000 eee eee A 9 ALS Wal harder ok eet e Rey ape atta etd A 10 ALO A ok ete se neat Seed boheme deed ae en es A
22. n a printout of the instructions select File Print in the Method editor and click on the Instruction set option box As an example this appendix lists the instructions for methods manual control and system settings supported by the strategy for AKTAexplorer 100 with Frac 950 E100F200 A 1 Method instructions A 1 1 Pump Instruction Description Parameters Flow Starts the flow for the gradi Flow rate ent pump at the set flow rate 0 100 ml min Gradient Defines a linear gradient for Target gradient pump pump A and 0 100 B B Reaches the target value Length after the specified length 0 9999 base Length is expressed in time volume or column volume depending on selected base BufferPrep_pH Selects pH for a defined Buff pH erPrep recipe When Buffer 0 14 Prep is not used the instruction should be set to zero otherwise an alarm will be given SampleFlow Starts the flow of the sample Flow rate pump at the set flow rate The 0 5 ml min sample pump can be cali brated in the calibration mod ule Al DirectLoad The sample is directly loaded SampleFlow onto the column ReduceFlow 0 50 makes it possible to automati Volume cally reduce the flow during 0 1000 sample loading to avoid a ReduceFlow pressure alarm An alarm will No be received if the flow is lower Yes than the flow specified in MinFlow MinFlow If an alarm is exe OFF 0 1 49 9 ml
23. n will deliver buffer with the selected composition Recipe and BufferPrep pH The sys tem is set to Pause during Sys temWash and the pressure limit is set to the value given in instruction Wash alarm pressure in System Set tings After System Wash the OutletValve and Column Valves turn back to their pre vious positions and the flow and pressure limit are set to their previous values Flow rate 25 ml min Wash vol ume 75 ml InletA1 A11 A18 MethodBase Sets the method volume calcu lation from either Gradient Pump or SamplePump Base GradientPump SamplePump um 18 1138 53AB e A4 A 1 2 Flowpath Instruction Description Parameters Injection Valve Sets the Injection Valve positions The Injection Valve is switched to position 1 with parameter load to position 2 with parameter inject and to position 3 with parameter waste The instruction gives an injection mark when the injection valve switches to the inject position Position Load Inject Waste ColumnPosition Selects the position for the column ColumnPosition switches the two col umn valves at the same time Position1Bypass corresponds to a tubing connected to valve port 1 and Position2 corresponds to a column connected to valve port 2 etc The flow rate is reduced to O ml min when the valves switch Position Position1 Bypass Position2 8 OutletValve Selects the position for t
24. ngs will be used Tube Next group change is delayed with respect to Tube number the delay volume To stop Peak Base Fractionation use the instruction Volume Peak_FracStop Time Parameters Tube type The tube type to frac tionate in PeakSize The size of the peak frac tions Start at Where to start the fraction ation if a specific position is desired use Tube number Base The peak fractionation can be performed in time or volume base Peak FracStop Stops the peak fraction collection after the set delay volume has been collected Peak Frac Parame UV1 Parameter settings for StartSlope ters Peak Fractionation To obtain suit 0 01 10000 able values perform a test run and mAu min measure the slope at the beginning EndSlope and end of the smallest and flattest 0 01 10000 peak you want to collect using mAU min Operations Differentiate in Eval MinWidth uation Time min as base must be 0 01 1500 used Minwidth prevents collection min of noise spikes Enter a value of about half the width of the narrow est peak to collect Reset_Frac_Position Resets the position of the fraction collector for small and large tubes and for start at next tube OutletFractions Enables fraction collection in subse How many quent outlets from the Outlet Valve 0 7 Fractionation starts at the selected Volume outlet If the number of pro 5 100000 ml grammed fractions exceeds the Star
25. on 6 tab within Run setup 4 Frac 950 tab within Documentation 7 fraction order options 5 fraction text alignment changing 8 illustrated 8 location 7 installation network B 2 PCI expansion card C 1 um 18 1138 53AB e ii stand alone B 2 instructions manual control A 13 pump A 1 system settings A 15 instructions manual control alarms and monitors A 14 flowpath A 13 frac A 13 miscellaneous A 15 pump A 13 instructions system settings alarms A 15 curve A 20 monitors A 18 specials A 17 introduction to UNICORN 3 L last tube illustration 6 options 5 setting default position 6 manual control instructions A 13 method instructions alarms and monitors A 9 flowpath A 4 Frac A 6 miscellaneous A 11 pump A 1 watch A 10 monitors system settings instructions A 18 N network installation B 2 P PCI expansion card installation C 1 pump manual control instructions A 13 pump instructions A 1 R rack type options 4 S specials system settings instructions A 17 stand alone installation B 2 strategy finding description of instruction A 1 for AKTAexplorer 100 A 1 system requirements hardware B 1 networks B 1 software B 1 system settings instructions A 15 T technical specifications B 1 typeface conventions 3 U UNICORN chromatography systems 3 introduction 3 Ww watch instructions A 10
26. on Parameters Fractionation Starts fraction collection if the frac TubeType tion size specified by the FracSize FracSize parameter is gt 0 Tube change is 0 000 delayed with respect to the set delay 99999 999 volume FractionCollector is base stopped with the instruction Frac StartAt tionationStop First tube Parameters Next tube Tube type The tube type to frac Next line tionate in Next group FracSize The size of the fractions Tube number Start at Where to start the fraction Base ation if a specific position is Volume desired use Tube number Time Base The fractionation can be per formed in time or volume base FractionationStop Stops fraction collecting after the set delay volume has been collected The flow is diverted to the waste of the outlet valve FeedTube During Fractionation or Peak Fractionation FeedTube moves the tube rack forward one tube after the set delay volume has been collected A fraction mark is given um 18 1138 53AB e A6 Peak Fractionation Peak_Fractionation enables only TubeType peaks to be collected The UV mon FracSize itor is used to determine when there 0 000 is a peak to collect PeakSize deter 99999 999 mines max volume to be collected base in each tube To collect the peak in StartAt one tube set PeakSize gt peak volume First tube If Peak_FracParameters are not set Next tube in the method default values in sys Next line tem setti
27. ovided for AKTAdesign systems and BioPilot System while configurations for process scale separation with BioProcess System may be customised for a particular installation UNICORN runs under the operating system Microsoft Windows NT version 4 0 This manual describes the functions of the fraction collector Frac 950 within UNICORN 3 20 For a detailed background on other UNICORN functions the user is referred to the UNICORN 3 10 manual for a detailed description of the Frac 950 functions see the Fraction collector Frac 950 user manual Functional features of Frac 950 include e User defined rack choice including tubes and microtubes e A variety of sampling orders during the fractionation process e Method templates providing method frameworks for fractionation applications eliminating the need to program methods from scratch For the purposes of this user manual examples are displayed for an AKTAexplorer 100 system You should refer to the user instructions specific to the chromatography system that you are using In this manual menu commands the names of dialogue boxes and windows the contents of dialogue boxes windows and option buttons are written with a bold helvetica typeface Menu commands are written in the order of the menu name and then the command separated by a colon For example Select File Save As to display the Save As dialogue Locate the destination drive and folder and enter a file name Click on S
28. t at F2 F8 number of outlets the fractionation will resume in position F3 After completing the last fraction the posi tion selected before executing Out letFractions instruction will be executed AccumulatorWash The accumulator makes 10 strokes OFF with 1 ml in each The flow must be ON gt 0 ml min When using the AccumulatorWash instruction for AKTAdesign systems use manual control to begin flow and turn the Outlet valve to the frac position um 18 1138 53AB e A8 A 1 4 Alarms amp Monitors Instruction Description Parameters Wavelength Sets the wavelength and number UV1 lof wavelengths to monitor The 190 700 range is 190 700 nm for UV1 nm UV2 UV2 and UV3 0 OFF for UV2 0FF and UV3 Wavelengths lt 190 nm 0 700 nm are automatically set to O off UV3 0 OFF 0 700 nm AutoZeroUV Sets the relative AU to zero Averaging TimeUV Filters the noise in the UV signal AvTimeUV Averaging time is the time inter 0 01 val used for calculating the mov 0 02 ing average of the absorbance 0 04 signal A long averaging time will 0 08 smooth out noise efficiently but 0 16 will also distort the peaks 0 32 Choose an averaging time of the 0 64 greatest value that can be set 1 28 which is less than 1 11 60 s 2 56 min typical peak width ml 5 12 flow rate ml min The averaging time should not be less than 1 28s when more than one wav
29. thod below the marked instruction Comment um 18 1138 53AB e A12 A 2 Manual control A 2 1 Pump This group contains the same instructions as the Pump group in method instructions Section A 1 1 A 2 2 Flowpath This group contains the same instructions as the Flowpath group in method instructions Section A 1 2 A 2 3 Frac This group contains the same instructions as the Flowpath group in method instructions section A 1 3 with the following instructions replacing the Frac instructions parameter is gt 0 Tube change is delayed with respect to the set delay volume FractionCollector is stopped with the instruction Frac tionationStop Parameter s Rack The rack used Frac_order The fractionation order kan be selected as Serpentine row Row by row Serpentine Column lor Column by column Tube type The tube type to frac tionate in FracSize The size of the fractions Start at Where to start the fraction ation if a specific position is desired use Tube number Base The fractionation can be per formed in time or volume base Instruction Description Parameters Man Fractionation Starts fraction collection if the frac Rack tion size specified by the FracSize Frac_order Serpentine row Row by row Serpentine column Column by col umn Tube type FracSize 0 000 99999 999 base StartAt First tube Next tube Next line Next group Tube number Base Volume
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