No 900 Mixer AKTA PURIFIER Manual
User Manual: 900 Mixer
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ÄKTA system Training Guide Purifier 100 / 10 ham am ers en biosci ham amers ce nce bioscie amersham bioscience am ersh am bi os ci en ce ÄKTApurifier Buffers Injector Box Monitor On-line Filter Conductivity pH Outlet Valve UV Detector Mixer Pumps Pump A Pump B Buffer switching valve ÄKTApurifier 10 / 100 In-line filter 1 7 6 2 3 4 5 Injection valve (V1) UV 3 Wavelength detector Cond. Conductivity monitor pH Mixer pH detector System pumps A Fraction collector Frac-950 B 1 8 2 3 4 7 6 5 Outlet valve (V4) ÄKTApurifier + Sample Pump + Sample Valve In-line filter Injection valve (V1) 1 7 6 2 3 45 Mixer UV 3 Wavelength detector Cond Conductivity monitor System pumps A pH B Sample pump pH detector Fraction collector Frac-950 1 87 2 6 345 1 87 2 6 345 Sample selection valve (V5) Outlet valve (V4) ÄKTApurifier 100 Air In-line filter 1 7 6 2 3 4 5 Injection valve (V1) UV 3 Wavelength detector Cond. Conductivity monitor pH Mixer pH detector System pumps A Fraction collector Frac-950 B Air sensor 8 1 2 3 4 7 6 5 Outlet valve (V4) ÄKTApurifier + Buffer Valve In-line filter 1 7 6 2 3 4 5 Injection valve (V1) UV 3 Wavelength detector Cond. Conductivity monitor pH Mixer pH detector System pumps A Fraction collector Frac-950 B 1 8 2 3 1 87 2 6 345 Buffer select valve (V6) 4 7 6 5 Outlet valve (V4) ÄKTApurifier 10 XT In-line filter 1 7 6 2 3 4 5 Injection valve (V1) UV 3 Wavelength detector Cond. Conductivity monitor pH Mixer pH detector System pumps A Fraction collector Frac-950 B 1 8 2 3 4 7 6 5 Outlet valve (V4) PUMP P-900 Mixer Pump A Pump B Proportion Valve A2 Acid /Alkali ÄKTA 10 mls/min ÄKTA 100 mls/min A1 B2 Buffer B1 Salt GRADIENT <6 mls/min 6 -14 mls/min <30 mls/min 30 - 50 mls/min 50 - 100 mls/min Water MIXER 0.6 ml mixer 2 ml mixer 2 mls mixer 5 mls mixer 12 mls mixer PUMP P-900 Inlets Mixer Pump B Pump A Switching Valves Buffer Select Valve IV-908 A2 A1 2 3 A11 A12 A13 1 8 4 A14 B1 Water 7 5 6 A16 A15 B2 A17 A18 Salt PUMP P-900 Manually Priming Buffer Inlet Lines Pump A Common inlet A2 Pump B A1 Common inlet Manual Flowpath Manual A1 or A2 BufferValveA B1 or B2 B1 B2 Flowpath BufferValveB Open Draw A ir out of Buffer Line PUMP P-900 Back - Wash Lines Pump Back-Wash Lines Syringe fitting Wash OUT IN-LINE one-way valve Buffer OUT Ball outlet check valve High pressure Piston seal Piston 20% Ethanol : Water Backwash pump diaphragm Ball inlet check valve Wash IN Buffer IN ON-LINE FILTERS P-903 ( 10 ml/min Pump ) Low Flow HPLC Filter P-901 ( 100 ml/min Pump ) High Flow Fibre Filter Polymer Support LOAD (sample) INJECT (sample) 2 3 2 Pump 7 4 4 6 5 8 4 4 Waste 6 3 4 5 Sample 8 7 6 3 6 5 2 2 5 6 PV-908 1 8 7 7 2 Pump 7 3 PV-908 1 Sample 2 Pump 7 3 6 5 PV-908 4 1 1 1 3 Column Column Column 1 LOAD (wash loop) Waste 5 Sample Waste INJECTION VALVE INV- 907 1 COLUMN 7 2 LOAD SAMPLE 3 PUMP 6 4 5 Waste Waste LOAD 1 COLUMN 7 2 PUMP 3 4 6 5 Waste Waste INJECT 1 COLUMN 7 2 PUMP 3 4 Waste 6 5 Waste WASTE SAMPLE LOADING METHOD 1 COLUMN 2 LOAD SAMPLE 7 PUMP 3 6 5 4 Needle Port Waste Waste Always tighten by hand with needle inserted Do Not Overtighten 1 COLUMN LOAD SAMPLE 7 2 PUMP 3 6 4 5 Luer Lock Adapter Waste Waste Superloop SUPERLOOPS 10 ml 50 ml Buffer 150 ml Plunger BUFFER FLUSH SAMPLE LOAD SAMPLE INJECT SAMPLE FLUSH WASTE PUMP or SYRINGE COLUMN COLUMN Sample Etched Groove Volume Max. allowed column pressure Code No. 10 ml 50 ml 150 ml 4 Mpa 4 Mpa 2 Mpa 18-1113-81 18-1113-82 18-1023-85 INV-907 Injection Valve - Injection Port PEEK Valve Body Injection Needle Port Valve Seal Ferrule NeedleTube Needle Needle Adapter INV-907 Injection Valve - Luer Adapter PEEK PEEKInjection InjectionValve ValveBody Body PEEK PEEKInjection InjectionLuer LuerAdaptor Adaptor Valve ValveSeal Seal Valve ValveChannel ChannelPlate Plate Fluid Dynamics Flow PEEK Tubing High Resistance to Flow Low Resistance to Flow End Beginning Fluid Dynamics Flow - Injecting a Loop The way we imagine it Flows The way it Actually Flows Bubble Segmentation 100ul Loop Waste 50ul Load 60ul Load 100ul Load 200ul Load • Partial Loading Loop = No more than 1/2 Loop Volume • Total Loading Loop = Inject 2X Loop Volume LOADING A SAMPLE LOOP Injection valve INV-907 1 Column 7 2 Load 3 Inject Sample 4 Sample pump P-950 Pump 6 5 Waste LOADING A SUPER LOOP Injection valve INV-907 1 Column 7 2 Load Inject 3 Pump 6 4 5 Buffer Sample Waste Sample MULTI-LOADING A SAMPLE LOOP Injection valve INV-907 1 Column 7 2 Load 3 Inject 4 Pump 6 5 PV-908 1 8 2 3 I/O 4 5 7 6 Wash Samples Waste MULTI-LOADING A SUPER LOOP Column 1 2 Pump Load Inject Injection valve INV-907 7 3 4 Waste 6 5 PV-908 8 1 2 3 I/O 4 5 7 6 Samples Wash S3 S4 S5 S6 S8 DIRECT LOADING ON TO COLUMN Column 1 Pump 7 2 3 Inject Load 6 4 5 Extra Tube 1/16” or 1/8” Tubing Waste Sample REVERSE FLOW VALVE ( INV-907 ) 1 7 2 Pump 3 5 4 Column Extra Tube 6 Stop Plug MONITOR Downward Flow Position 1 1 7 2 Pump 3 4 Column Extra Tube 5 6 Stop Plug MONITOR Upward Flow Position 2 OUTLET VALVE PV-908 DETECTOR To Fraction Collector Default F8 F1 F2 F3 I/O F4 F5 F7 F6 F7 Waste F8 F4 F3 F5 F6 Fraction Collection Flowthrough Fractionation Peak Fractionation Eluate Fractionation 100%B 0%B mAU 100 50 Waste End D N U O B N U D N U O B Start Inject 500 250 %B 0 A1 A2 A3…………………. 0 HiTrap Connections 1/16” Female 1/16” Male 18-1112-55 1/16” Fingertight Fitting 1/16” Female M6 Male 18-1112-57 M6 male / 1/16” Female Phenyl Sepharose HiTrap Column M6 Male M6 Female 18-1112-58 M6 female / 1/16” male Flow Cell Inlet 1/16” Male Multiple HiTrap Arrangment ( Series ) Phenyl Sepharose Phenyl Sepharose Phenyl Sepharose Phenyl Sepharose Phenyl Sepharose Phenyl Sepharose De or t c te t or c e t De or t c te De Traditional FPLC fitting to ÄKTA / HPLC Injector 1/16” Fingertight Fitting 18-1112-55 10 0u 1/16” FEMALE / M6 FEMALE 18-1123-94 l Flange M6 Fitting 1/16” MALE / M6 FEMALE 18-1112-58 Flow Cell Tubing Fittings 18-1112-55 1/16” Fingertight Fitting 1/16” PEEK TM Tubing 18-1112-58 Flange Fitting Adapter FPLC Flange Fitting 18-1112-51 Luer Fitting Adapter 18-1112-52 Fingertight Stop Plug Flangless Inlet Connector 3/16” 18-1112-49 1/8” 18-1121-17 1/16” 18-1127-07 18-1112-57 M6 male / 1/16” Female Luer Syringe 18-1112-50 5/16” Stop Plug Flangless Buffer Inlet Ferrule 3/16” 18-1112-48 1/8” 18-1121-18 1/16” 18-1127-06 18-1017-98 Tubing Connector M6 male / O - ring 18-3955-01 1/16”female X 1/16”female Union Ti Union + 2 Fingertight Fittings 18-1123-94 M6 female X 1/16”female Union FPLC Flange - 1/16” Fingertight COMPRESSION FITTINGS Untightened Correctly tightened Obstructed Over tightened Tubing Restricted PEEK Tubing TM Gray, 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 Tubing 3/16” OD 18-1112-47 1/8” OD 18-1121-16 1/16” OD 18-1119-74 Tubing Kit Changes - ÄKTApurifier 10 GENERAL FLOW (at delivery) G5 10 0u l Injection valve V1 G9 Outlet Valve V4 (7ul) Accumulator (40ul) UV Flow cell G8 G6 0.25 mm i.d Blue PEEK G5 400mm G6 160mm G7 80mm G8 130mm G9 500mm G7 Conductivity Flow cell (14ul) FR-904 (0.2Mpa, 5ul) NB: Never use the FR-904 with the pH electrode pH Flow cell (100ul) Tubing Kit AKTApurifier 10 , 0.50 tubing kit AKTApurifier 10 , 0.50 tubing kit + pH electrode AKTApurifier 10 , 0.25 tubing kit No Frac Frac 901 232 354 88 Frac 950 382 504 238 Tubing Kit Changes - ÄKTApurifier 10 HIGH FLOW G5 10 0u l Injection valve V1 H9 Outlet Valve V4 (7ul) Accumulator (40ul) UV Flow cell H8 H6 H7 Conductivity Flow cell (14ul) 0.50 mm i.d Orange PEEK H6 160mm H7 80mm H8 130mm H9 500mm FR-904 (0.2Mpa, 5ul) NB: Never use the FR-904 with the pH electrode pH Flow cell (100ul) Tubing Kit AKTApurifier 10 , 0.50 tubing kit AKTApurifier 10 , 0.50 tubing kit + pH electrode AKTApurifier 10 , 0.25 tubing kit No Frac Frac 901 232 354 88 Frac 950 382 504 238 Tubing Kit Changes - ÄKTApurifier 100 HIGH FLOW (at delivery) G5 10 0u l Injection valve V1 G9 Outlet Valve V4 (7ul) Accumulator (40ul) UV Flow cell G8 G6 0.75 mm i.d Green PEEK G6 G7 G8 G9 G7 Conductivity Flow cell (14ul) FR-904 (0.2Mpa, 5ul) 160mm 80mm 130mm 500mm NB: Never use the FR-904 with the pH electrode pH Flow cell (100ul) Tubing Kit AKTApurifier 100 , 0.50 tubing kit AKTApurifier 100 , 0.75 tubing kit AKTApurifier 100 , 1.00 tubing kit AKTApurifier 100 , 0.50 tubing kit + pH electrode AKTApurifier 100 , 0.75 tubing kit + pH electrode AKTApurifier 100 , 1.00 tubing kit + pH electrode AKTAbasic 100 , 0.75 tubing No Frac 290 Frac 901 232 470 769 354 672 955 Frac 950 382 620 809 504 862 995 Tubing Kit Changes - ÄKTApurifier 100 LOW FLOW L5 10 0u l Injection valve V1 L9 Outlet Valve V4 (7ul) Accumulator (40ul) UV Flow cell L8 L6 0.50 mm i.d Orange PEEK L6 L7 L8 L9 L7 Conductivity Flow cell (14ul) 160mm 80mm 130mm 500mm FR-904 (0.2Mpa, 5ul) NB: Never use the FR-904 with the pH electrode pH Flow cell (100ul) Tubing Kit AKTApurifier AKTApurifier AKTApurifier AKTApurifier AKTApurifier AKTApurifier No Frac 100 100 100 100 100 100 , , , , , , 0.50 0.75 1.00 0.50 0.75 1.00 tubing tubing tubing tubing tubing tubing kit kit kit kit + pH electrode kit + pH electrode kit + pH electrode Frac 901 232 470 769 354 672 955 Frac 950 382 620 809 504 862 995 A-900 Autosampler INV-907 1 A-900 2 1 2 7 6 3 4 5 3 MANUAL SAMPLE LOAD LOAD 6 5 4 Waste COLUMN Sample PUMP Waste LOAD Syringe INV-907 1 A-900 2 1 6 3 4 5 3 MANUAL SAMPLE LOAD INJECT Syringe Sample 7 2 4 6 5 Waste Waste COLUMN PUMP LOAD Needle Height Standard Vial Standard Vial + Needle Height Insert volume left 2mm 450ul 1mm 350ul 0mm 200ul 3mm 12ul A-900 Sample Injection ( Flushed Loop Injection ) INJECT INJECT INJECT INJECT LOAD LOAD pump waste INJECT INJECT pump waste waste waste column column pump pump column column Sample needle sample INJECT,sample sample InInINJECT, flushesthe thevalve valve flushes channel. channel. Min.( (30ul 30ul) )with with Min. AirSegment Segment Air Min.( (35ul 35ul) )without without Min. Airsegment segment Air sample LOADThe Thesample sample InInLOAD drawnup upto to isisdrawn completely completely fillthe theloop. loop. fill noless lessthan thantwice twice ( (no loopvolume volume loop recommended) ) recommended Wash Reservoir sample INJECTsample sample InInINJECT washedonto onto isiswashed thecolumn. column. the sample Finalinternal internal Final needlewash. wash. needle pump waste column INJECT INJECT OptionalExternal External Optional needlewash. wash. needle sample transport fluid A-900 Sample Injection ( Partial Loop Injection ) INJECT INJECT INJECT INJECT LOAD LOAD INJECT INJECT waste pump pump waste waste sample INJECT,sample sample InInINJECT, flushesthe thevalve valve flushes channel. channel. Min.( (30ul 30ul) )with with Min. AirSegment Segment Air Min.( (35ul 35ul) )without without Min. Airsegment segment Air waste waste column Sample needle pump pump column Sample needle sample Thesample sampleisisdrawn drawn The upto topartially partiallyfill fillthe the up loopwith with loop programmedvolume. volume. programmed nomore morethan thanhalf half ( (no loopvolume volume loop recommended) ) recommended column column Sample needle sample injectsample sample InIninject washedonto onto isiswashed thecolumn. column. the Wash Reservoir sample Finalinternal internal Final needlewash. wash. needle pump waste column INJECT INJECT OptionalExternal External Optional needlewash. wash. needle sample transport fluid A-900 Sample Injection ( ul Pick-up Injection ) INJECT INJECT LOAD LOAD pump pump waste transport fluid sample sample Thesample sampleisis The drawnup upfrom from drawn thesample samplevial vial the INJECT INJECT column Sample needle Sample needle sample sample Thesample sampleisis The washedinto into washed theloop loopwith with the transportfluid fluid transport pump injectsample sample InIninject washedonto ontothe theco column isiswashed lumn pump waste waste Finalinternal internal Final needlewash. wash. needle Wash Reservoir pump column Sample needle inject,valve valve InIninject, flusheswith with flushes transportfluid fluid transport pump column column INJECT INJECT LOAD LOAD column sample column INJECT INJECT Optionalneedle needle Optional wash. wash. sample transport fluid System Compensation Volume Equilibration Phase A B COLUMN TOP 0% A B COLUMN TOP 20% Effect 8 mls Command 20%B 0%B 8mls Gradient Delay Volume Gradient Phase A B COLUMN TOP 0% A B COLUMN TOP 100% Effect 16 mls 100%B Gradient Delay Command 100% Column Pressure ( compensation ) Flow Restrictor Column Pressures Column Hardware Pressure Limits Column Shatters DANGEROUS Column Matrix Pressure Limits Matrix Collapses EXPENSIVE 0.3Reading Reading 0.3 Atmos Atmos Press Press 0.0MPa MPa 0.0 Pre-column pressure = 0 MPa Pump Pump 0.3Mpa Mpa 0.3 0.3 MPa GELPress PressDrop Drop GEL 0.3MPa MPa 0.3 (0.3- -0.0) 0.0) (0.3 0.0 MPa 0.0MPa MPa 0.0 0.5Reading Reading 0.5 Pre-column pressure = 0 MPa Gel Top Pressure Pump Pump 0.3Mpa Mpa 0.3 0.3 MPa ColumnPress PressDrop Drop Column GELPress PressDrop Drop GEL Colu mnPress PressDrop Drop Colu mn (0.3++0.0) 0.0) (0.3 (0.3- -0.2) 0.2) (0.3 (0.5++0.2) 0.2) (0.5 0.3MPa MPa 0.3 Gel Bottom Pressure 0.1MPa MPa 0.1 0.2 MPa 0.2MPa MPa 0.2 FlowRestrictor Restrictor Flow 0.5MPa MPa 0.5
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