Transcription
ÄKTA system Training GuidePurifier 100 / 10hamam ersenbioscihamamerscencebioscieamersham bioscienceamershambi osci ence
rConductivitypHOutlet ValveUV DetectorMixerPumpsPump APump BBuffer switchingvalve
ÄKTApurifier 10 / 100In-linefilter1 7623 4 5Injectionvalve (V1)UV3 H 4765Outletvalve (V4)ÄKTApurifier Sample Pump Sample ValveIn-linefilterInjectionvalve (V1)1 7623 45MixerUV3 psApHBSamplepumppH detectorFractioncollectorFrac-9501 87263451 8726345Sample selectionvalve (V5)Outletvalve (V4)
ÄKTApurifier 100 AirIn-linefilter1 7623 4 5Injectionvalve (V1)UV3 H ensor81234765Outletvalve (V4)ÄKTApurifier Buffer ValveIn-linefilter1 7623 4 5Injectionvalve (V1)UV3 H 1 8726345Bufferselectvalve (V6)4765Outletvalve (V4)
ÄKTApurifier 10 XTIn-linefilter1 7623 4 5Injectionvalve (V1)UV3 H 4765Outletvalve (V4)
PUMP P-900MixerPump APump BProportionValveA2Acid /AlkaliÄKTA 10 mls/minÄKTA 100 mls/minA1 B2BufferB1SaltGRADIENT 6 mls/min6 -14 mls/min 30 mls/min30 - 50 mls/min50 - 100 mls/minWaterMIXER0.6 ml mixer2 ml mixer2 mls mixer5 mls mixer12 mls mixer
PUMP P-900 InletsMixerPump BPump ASwitchingValvesBuffer SelectValve IV-908A2A123A11A12A131 84A14B1Water756A16A15B2A17A18Salt
PUMP P-900 Manually Priming Buffer Inlet LinesPump ACommon inletA2Pump BA1Common inletManualFlowpathManualA1 or A2BufferValveA B1 or B2B1B2FlowpathBufferValveBOpenDraw Airout ofBufferLine
PUMP P-900 Back - Wash LinesPump Back-WashLinesSyringe fittingWash OUTIN-LINEone-wayvalveBufferOUTBall outletcheck valveHigh pressurePiston sealPiston20% Ethanol : WaterBackwash pumpdiaphragmBall inletcheck valveWashINBufferIN
ON-LINE FILTERSP-903 ( 10 ml/min Pump )Low Flow HPLC FilterP-901 ( 100 ml/min Pump )High Flow Fibre FilterPolymer SupportLOAD (sample)INJECT lumnColumnColumn1LOAD (wash loop)Waste5SampleWaste
INJECTION VALVE INV- asteWASTE
SAMPLE LOADING METHOD1COLUMN2LOADSAMPLE7PUMP3654Needle PortWasteWasteAlways tighten by handwith needle insertedDo Not Overtighten1COLUMNLOADSAMPLE72PUMP3645Luer LockAdapterWasteWasteSuperloop
SUPERLOOPS10 ml50 mlBuffer150 USHWASTEPUMP orSYRINGECOLUMNCOLUMNSampleEtchedGrooveVolumeMax. allowed columnpressureCode No.10 ml50 ml150 ml4 Mpa4 Mpa2 Mpa18-1113-8118-1113-8218-1023-85
INV-907 Injection Valve - Injection PortPEEK Valve BodyInjection Needle PortValve SealFerruleNeedleTubeNeedleNeedleAdapter
INV-907 Injection Valve - Luer e
Fluid DynamicsFlowPEEK TubingHigh Resistanceto FlowLow Resistanceto FlowEndBeginning
Fluid DynamicsFlow - Injecting a LoopThe way we imagine it FlowsThe way it Actually FlowsBubble Segmentation100ul LoopWaste50ul Load60ul Load100ul Load200ul Load Partial Loading Loop No more than 1/2 Loop Volume Total Loading Loop Inject 2X Loop Volume
LOADING A SAMPLE LOOPInjection -950Pump65Waste
LOADING A SUPER LOOPInjection eWasteSample
MULTI-LOADING A SAMPLE LOOPInjection /O4576WashSamplesWaste
MULTI-LOADING A SUPER LOOPColumn12PumpLoadInjectInjection S3 S4 S5 S6S8
DIRECT LOADING ON TO COLUMNColumn1Pump723InjectLoad645Extra Tube1/16” or 1/8”TubingWasteSample
REVERSE FLOW VALVE ( INV-907 )172Pump354ColumnExtra Tube6StopPlugMONITORDownward FlowPosition 1172Pump34ColumnExtra Tube56StopPlugMONITORUpward FlowPosition 2
OUTLET VALVE PV-908DETECTORTo F8F4F3F5F6
Fraction UDNUOBStartInject500250%B0A1 A2 A3 .0
HiTrap Connections1/16” Female1/16” Male18-1112-551/16” Fingertight Fitting1/16” FemaleM6 Male18-1112-57M6 male / 1/16” FemalePhenyl SepharoseHiTrap ColumnM6 MaleM6 Female18-1112-58M6 female / 1/16” maleFlow Cell Inlet1/16” Male
Multiple HiTrap Arrangment ( Series )Phenyl SepharosePhenyl SepharosePhenyl SepharosePhenyl SepharosePhenyl SepharosePhenyl SepharoseDeortctet orcetDeortcteDe
Traditional FPLC fitting to ÄKTA / HPLCInjector1/16” Fingertight Fitting18-1112-55100u1/16” FEMALE / M6 FEMALE18-1123-94lFlange M6 Fitting1/16” MALE / M6 FEMALE18-1112-58Flow Cell
Tubing Fittings18-1112-551/16” Fingertight Fitting1/16” PEEKTMTubing18-1112-58Flange Fitting AdapterFPLC Flange Fitting18-1112-51Luer Fitting Adapter18-1112-52Fingertight Stop PlugFlangless Inlet 8-1127-0718-1112-57M6 male / 1/16” FemaleLuer Syringe18-1112-505/16” Stop PlugFlangless Buffer ��18-1127-0618-1017-98Tubing ConnectorM6 male / O - ring18-3955-011/16”female X 1/16”female Union TiUnion 2 Fingertight Fittings18-1123-94M6 female X 1/16”female UnionFPLC Flange - 1/16” Fingertight
COMPRESSION FITTINGSUntightenedCorrectly tightenedObstructedOver tightened Tubing Restricted
PEEK TubingTMGray, 1/16”x 1.0 mm ( 7.85ul / cm )Green, 1/16”x 0.75 mm ( 4.42ul / cm )Orange, 1/16”x 0.5 mm ( 1.96ul / cm )Blue, 1/16”x 0.25 mm ( 0.49ul / cm )Teflon / Tefzel Tubing3/16” OD18-1112-471/8” OD18-1121-161/16” OD18-1119-74
Tubing Kit Changes - ÄKTApurifier 10GENERAL FLOW (at delivery)G5100ulInjection valve V1G9Outlet Valve V4 (7ul)Accumulator(40ul)UV Flow cellG8G60.25 mm i.dBlue PEEKG5 400mmG6 160mmG7 80mmG8 130mmG9 500mmG7ConductivityFlow cell (14ul)FR-904(0.2Mpa, 5ul)NB: Never usethe FR-904 withthe pH electrodepH Flow cell (100ul)Tubing KitAKTApurifier 10 , 0.50 tubing kitAKTApurifier 10 , 0.50 tubing kit pH electrodeAKTApurifier 10 , 0.25 tubing kitNo FracFrac 90123235488Frac 950382504238
Tubing Kit Changes - ÄKTApurifier 10HIGH FLOWG5100ulInjection valve V1H9Outlet Valve V4 (7ul)Accumulator(40ul)UV Flow cellH8H6H7ConductivityFlow cell (14ul)0.50 mm i.dOrange PEEKH6 160mmH7 80mmH8 130mmH9 500mmFR-904(0.2Mpa, 5ul)NB: Never usethe FR-904 withthe pH electrodepH Flow cell (100ul)Tubing KitAKTApurifier 10 , 0.50 tubing kitAKTApurifier 10 , 0.50 tubing kit pH electrodeAKTApurifier 10 , 0.25 tubing kitNo FracFrac 90123235488Frac 950382504238
Tubing Kit Changes - ÄKTApurifier 100HIGH FLOW (at delivery)G5100ulInjection valve V1G9Outlet Valve V4 (7ul)Accumulator(40ul)UV Flow cellG8G60.75 mm i.dGreen PEEKG6G7G8G9G7ConductivityFlow cell (14ul)FR-904(0.2Mpa, 5ul)160mm80mm130mm500mmNB: Never usethe FR-904 withthe pH electrodepH Flow cell (100ul)Tubing KitAKTApurifier 100 , 0.50 tubing kitAKTApurifier 100 , 0.75 tubing kitAKTApurifier 100 , 1.00 tubing kitAKTApurifier 100 , 0.50 tubing kit pH electrodeAKTApurifier 100 , 0.75 tubing kit pH electrodeAKTApurifier 100 , 1.00 tubing kit pH electrodeAKTAbasic 100 , 0.75 tubingNo Frac290Frac 901232470769354672955Frac 950382620809504862995
Tubing Kit Changes - ÄKTApurifier 100LOW FLOWL5100ulInjection valve V1L9Outlet Valve V4 (7ul)Accumulator(40ul)UV Flow cellL8L60.50 mm i.dOrange PEEKL6L7L8L9L7ConductivityFlow cell (14ul)160mm80mm130mm500mmFR-904(0.2Mpa, 5ul)NB: Never usethe FR-904 withthe pH electrodepH Flow cell (100ul)Tubing rAKTApurifierAKTApurifierNo 00tubingtubingtubingtubingtubingtubingkitkitkitkit pH electrodekit pH electrodekit pH electrodeFrac 901232470769354672955Frac 950382620809504862995
A-900 465WasteWasteCOLUMNPUMPLOAD
Needle HeightStandard VialStandard Vial Needle Height Insertvolume left2mm450ul1mm350ul0mm200ul3mm12ul
A-900Sample Injection ( Flushed Loop Injection olumnSample hethevalvevalveflusheschannel.channel.Min.( (30ul30ul) )withwithMin.AirSegmentSegmentAirMin.( (35ul35ul) sthanthantwicetwice( (noloopvolumevolumelooprecommended) ortfluid
A-900Sample Injection ( Partial Loop Injection Min.( (30ul30ul) )withwithMin.AirSegmentSegmentAirMin.( (35ul35ul) olumnSample needlepumppumpcolumnSample anhalfhalf( (noloopvolumevolumelooprecommended) )recommendedcolumncolumnSample id
A-900Sample Injection ( ul Pick-up Injection mple needleSample ReservoirpumpcolumnSample ortfluid
System Compensation VolumeEquilibration PhaseABCOLUMN TOP0%ABCOLUMN TOP20%Effect8 mlsCommand20%B0%B8mls
Gradient Delay VolumeGradient PhaseABCOLUMN TOP0%ABCOLUMN TOP100%Effect16 mls100%BGradient DelayCommand 100%
Column Pressure ( compensation ) Flow RestrictorColumn PressuresColumn Hardware Pressure LimitsColumn Shatters DANGEROUSColumn Matrix Pressure LimitsMatrix Collapses .0MPaMPa0.0Pre-column pressure 0 MPaPumpPump0.3MpaMpa0.30.3 MPaGELPressPressDropDropGEL0.3MPaMPa0.3(0.3- -0.0)0.0)(0.30.0 MPa0.0MPaMPa0.00.5ReadingReading0.5Pre-column pressure 0 MPaGel Top PressurePumpPump0.3MpaMpa0.30.3 DropGELColumnPressPressDropDropColumn(0.3 0.0)0.0)(0.3(0.3- -0.2)0.2)(0.3(0.5 0.2)0.2)(0.50.3MPaMPa0.3Gel Bottom Pressure0.1MPaMPa0.10.2 Pa0.5
AKTApurifier 100 , 0.50 tubing kit 232 382 AKTApurifier 100 , 0.75 tubing kit 470 620 AKTApurifier 100 , 1.00 tubing kit 769 809 AKTApurifier 100 , 0.50 tubing kit pH electrode 354 504 AKTApurifier 100 , 0.75 tubing kit pH electrode 672 862 AKTApurifier 100 , 1.00 tubing kit pH electrode 955 995 AKTAbasic 100 , 0.75 tubing 290
