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Amersham Biosciences AKTA design A-905 User Manual

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Autosampler A-905
User Manual
18-1154-42

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Summary of Contents for Amersham Biosciences AKTA design A-905

  • Page 1 Autosampler A-905 User Manual 18-1154-42...
  • Page 3 Unless otherwise agreed in writing, all goods and services are sold subject to the terms and conditions of understand the safe use of Autosampler A-905. sale of the company within the Amersham Biosciences group which supplies them. A copy of these terms and WARNING! conditions is available on request.

  • Page 5: Table Of Contents

    Contents Contents 1 Introduction 1.1 General ..................7 1.2 Safety ..................8 1.3 Safety symbols on Autosampler A-905 ......... 8 2 Installation 2.1 Unpacking ................... 9 2.2 Locating the A-905 ..............10 2.3 Power connections ..............11 2.4 UniNet connections ..............11 2.5 Selecting A-905 in UNICORN ............
  • Page 6 Contents About this manual This manual comprises two parts; a practical part (sections 1–5) and a reference part (section 6). Sections 1–5 contain the necessary information for operating the instrument. Autosampler A-905 User Manual 18-1154-42 Edition AB...

  • Page 7: Introduction

    Introduction 1 Introduction 1.1 General ™ This manual describes Autosampler A-905 for ÄKTA ™ Autosampler A-905 for Ettan . They are automated sampling injectors for use in ÄKTA and Ettan liquid chromatography systems. The autosampler features: • Sample injection volumes as follows: •...

  • Page 8: Safety

    Introduction 1.2 Safety • The instrument is designed for indoor use only. • Do not use in a dusty atmosphere or close to spraying water. • Do not block the air inlet and outlet of the unit. WARNING! Always disconnect the power supply before attempting to replace any item on the instrument.

  • Page 9: Installation

    Installation 2 Installation 2.1 Unpacking Unpack the instrument and check the items against the supplied packing list. Inspect the items for obvious damage that may have occurred during transportation. Keep all packing materials if onward transport of the instrument is expected.

  • Page 10: Locating The A-905

    Installation 2.2 Locating the A-905 The A-905 needs approximately 28 cm of bench space and a mains connection of 220-240 V or 100-120 V, 50/60 Hz, preferably taken from the system base platform. Make sure that none of the ventilation holes are blocked. Blockage of the ventilation holes can cause malfunctioning of the A-905 Autosampler or even damage the electronics.

  • Page 11: Power Connections

    Installation 2.3 Power connections Before connecting the mains cord to your separation unit, check that the voltage setting of the A-905 matches the local mains supply voltage and main fuses. Use only a supply appliance with protective grounding. If the indicated voltage is not correct, select the proper voltage by removing, inverting, and then re-inserting the voltage selector- cartridge.

  • Page 12: Fluid Connections

    Installation 2.6 Fluid connections Autosampler A-905 for ÄKTA and Autosampler A-905 for Ettan are factory-installed with different dimensions on some of the fluid handling parts (see Table 2-1 ). Fluid handling part Autosampler A-905 Autosampler A-905 for ÄKTA for Ettan Syringe 1 ml 100 µl...
  • Page 13 Installation Tubing Material Dimensions × × Sample needle tubing PEEK 135 mm o.d. 0.5 mm i.d. 0.25 mm (Volume: 15 µl) × × Buffer tubing from high Teflon 2500 mm o.d. 1/16" i.d. 1.0 mm pressure valve to syringe valve (Volume: 2 ml) ×...
  • Page 14 Installation 2.6.1 System connections To ensure reproducible injections, make the connections to your chromatography system according to the installation instructions, given in the system documentation. Route the tubings through the tubing guide on the right-hand side. The A-905 has been flushed with 20% isopropanol.
  • Page 15 Installation 2.6.3 Wash solvent The A-905 has a built-in wash solvent reservoir of 250 ml (ÄKTA) or 100 ml Wash solvent (Ettan). tubing To position the wash solvent bottle: Fill the wash solvent bottle with 20% ethanol. Screw the bottle in the wash solvent holder and place the holder on the A-905.
  • Page 16 Installation After the wash solvent tubing and the syringe are filled, use the command in AutosamplerControl_905:InitialWash_905 to perform a standard wash. All tubing connected Manual:Flowpath to the syringe valve will be filled and flushed with wash solvent. The wash solvent bottle is now ready for use. If you have an application that requires more than 100 ml of wash solvent for a complete run, you can replace the wash solvent tubing by a longer tubing with one end flanged for the valve fitting and place a...

  • Page 17: Preparation Of Microplates And Vials

    Installation 2.7 Preparation of microplates and vials Autosampler A-905 is delivered with the following standard type microplates and transport vials: • 96-well low microplate (1 pc). • 96-well high microplate (1 pc). • 384-well low microplate (1 pc). • 48-vial adapter (1 pc). •...
  • Page 18 Installation 1 2 3 4 5 6 7 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 384-well microplate 48-vial adapter 96-well microplate (High and Low) Fig. 2-2. Microplate types for A-905 Autosampler A-905 User Manual 18-1154-42 Edition AB...
  • Page 19 Installation 2.7.2 Recommended microplates and transport vials The following 96-well microplates are recommended: Greiner nos., low microplates: 655101, 651101, 650101 Greiner nos., high microplates: 780201 Nunc nos., low microplates: 143761, 168055, 156545, 163320 The following transport vials are recommended: Chromacol no: 6 CV (6 ml), 10 CV (10 ml) 2.7.3 Filling and sealing wells and vials The wells, the standard vials, as well as the conical vials, are filled by...
  • Page 20 Installation 2.7.4 Initializing the microplate The plate type used must be selected in UNICORN. , select System Control System:Settings... Select in the field, and then Specials Instructions AutosamplerParameters_905:PlateType The instruction name does not contain “_905” in Autosampler for Ettan. Autosampler A-905 User Manual 18-1154-42 Edition AB...

  • Page 21: Operation

    Operation 3 Operation 3.1 General Switch on/off A-905 with the mains switch on the rear panel. A-905 is controlled from a PC running UNICORN version 4.12 or higher. It cannot be used as a stand-alone module. A-905 can be controlled, either automatically from a pre-made method, or manually via instructions.
  • Page 22 Operation 3.2.1 InjectionPickup_905 instruction selects the µl pick-up injection InjectionPickup_905 mode for A-905. The syringe dispenser aspirates transport liquid from the transport vial into the sample line. The A-905 injection valve is turned to LOAD position. The needle moves from the transport vial to a sample vial and aspirates the selected injection volume.
  • Page 23 Operation 3.2.2 InjectionFlushed_905 instruction selects the flushed loop injection InjectionFlushed_905 mode for A-905. The syringe dispenser aspirates a flush volume of sample into the sample line. The A-905 injection valve is turned to LOAD position, and the sample loop is filled with 1.5–3 times the sample loop volume (depending on the loop volume).

  • Page 24: Manual Instructions And Parameters

    Operation 3.3 Manual instructions and parameters 3.3.1 AutosamplerControl_905 This instruction contains manual commands used for setting up and maintaining Autosampler A-905. The available commands are: • InitialWash_905 • SyringeHome_905/SyringeEnd_905 • PlateHome_905/PlateExchange_905 • NeedleHome_905/NeedleFront_905 The commands are found by selecting in the Manual:Flowpath System module, and clicking...
  • Page 25 Operation PlateHome_905/EPlateExchange_905 commands are used when the PlateHome_905 /PlateExchange_905 plate is replaced. moves the plate to its normal position. PlateHome_905 moves the plate to the leftmost position. PlateExchange_905 NeedleHome_905/NeedleFront_905 commands are used when the NeedleHome_905 /NeedleFront_905 sample needle is replaced. moves the needle to its normal position.
  • Page 26 Operation 3.3.3 AutosamplerParameters_905 This instruction contains hardware related parameters for A-905. Parameter Value Comment LoopVol 5-1000 µl Sample loop volume 1 ml (ÄKTA) and 20 µl (Ettan) are default SyringeVol 100, 250, 500 or 1000 µl Syringe volume 1 ml (ÄKTA) and 100 µl (Ettan) are default FlushVol 10-100 µl...
  • Page 27 Operation 3.3.4 AutoSampler_Temperature_905 instruction is used to enable the AutoSampler_Temperature_905 cooling function and to set the desired temperature. The cooling function is enabled by selecting the check button. Enabled The cooling capacity is maximum 12 °C below the ambient < <...
  • Page 28 Operation 3.3.7 QuantitationData_905 instruction is used to send parameters to the Quantitation_Data_905 evaluation module when creating or updating the quantitation table. Different levels of standards are used for creating and updating the quantitation table. is used for quantification of the sample. Sample The instruction is found by selecting in the...

  • Page 29: Maintenance

    Maintenance 4 Maintenance 4.1 Injection valve 4.1.1 Description The injection valve is a two position, six-port valve. The figure shows the flow diagram of the valve. The six small circles represent the ports in the Shaft assembly Shaft assembly rotor seal. Rotating the valve rotated clockwise rotated counterclockwise 60°...
  • Page 30 Maintenance 4.1.3 Maintenance (Autosampler A-905 for ÄKTA) With normal use, the valve will give many tens of thousands of cycles of trouble-free operation. The main cause of early failure, which is seen as a leak in the valve, is abrasive particles in the sample and/or mobile phase scratching the rotor seal.
  • Page 31 Maintenance 4.1.4 Maintenance (Autosampler A-905 for Ettan) With normal use, the valve will give many tens of thousands of cycles of trouble-free operation. The main cause of early failure, which is seen as a leak in the valve, is abrasive particles in the sample and/or mobile phase scratching the rotor surface.
  • Page 32 Maintenance 4.1.5 Operating suggestions CAUTION! Never operate the injection valve when it is dry. This will reduce the lifetime of the injection valve and probably cause malfunction. CAUTION! Never leave the A-905 for more than one day with buffer solution in the tubings. Rinse the A-905 with distilled water prior to long stops, and fill the tubing with 20% ethanol.

  • Page 33: Needle And Tubing

    Maintenance 4.2 Needle and tubing The A-905 has separate needles for puncturing and sampling; the air needle, which punctures the septum of the vial and provides the headspace pressure in the vial, and the sample needle for transporting the sample from the vial into the loop of the valve. The sample needle enters a vial as follows: Fig.

  • Page 34: Syringe, Sample Loop And Buffer Tubing Combinations

    Maintenance 4.3 Syringe, sample loop and buffer tubing combinations Four sizes of dispenser syringes are available: 100, 250, 500 and 1000 µl. 4.3.1 Injection volumes Table 4-1 shows the injection volume ranges that are available for the the different injection modes. Injection mode Autosampler A-905 for ÄKTA Autosampler A-905 for Ettan...
  • Page 35 Maintenance 4.3.3 Flush volumes The recommended minimum flush volumes are 30 µl (ÄKTA) and 10 µl (Ettan). The default values for the flush volume are 40 µl (ÄKTA) and 10 µl (Ettan). The recommended flush volume with an air segment is 35 µl. Note: When an air segment is selected from the InjectionPickup_905 instruction in...
  • Page 36 Maintenance 4.3.5 Injection volumes smaller than 20 µl Use a 100 µl syringe and a 5 µl sample needle to obtain maximum reproducibility and accuracy. Use a 20 µl sample loop to avoid loss of accuracy due to expansion of the loop content when switching from inject to load position prior to sample loading.
  • Page 37 Maintenance 4.3.7 For volumes larger than 200 µl Select the syringe size according to the recommendation below: Syringe volume > 2 × injection volume. Injection volumes larger than 500 µl are possible, but the sample might contaminate the syringe. Program sufficient wash after use! Follow the recommendations in the table below for choosing the appropriate buffer tubing and sample loop: Syringe...

  • Page 38: Replacing The Sample Needle

    Maintenance 4.4 Replacing the sample needle To replace the sample needle, proceed as follows: 1 Use the AutosamplerControl_905:NeedleFront_905 command in to move the needle to the Manual:Flowpath front. Rheodyne nut 2 Loosen the needle connection nut (Male nut M6) that and ferrule fixes the sample needle.

  • Page 39: Syringe

    Maintenance 4.5 Syringe Autosampler A–905 is factory-equipped with a 1 ml syringe (ÄKTA) or a 100 µl syringe (Ettan). 250 µl and 500 µl syringes can also be used. To install another syringe, proceed as follows: Note: Do not disconnect the power supply of the A–905. It is needed to move the syringe.

  • Page 40: Troubleshooting

    5.1 Faults and actions Note: The instruction names does not contain “_905” in Autosampler for Ettan. If the suggested actions do not correct the fault, call your local Amersham Biosciences representative. Fault Possible cause Corrective action Bad reproducibility Injection valve switched dry...
  • Page 41 Troubleshooting Fault Possible cause Corrective action Syringe action failed, motor Syringe not tightened enough Tighten syringe generates noise when replaced Sample needle bent. No Septa type wrong, septa Replace needle and readjust injection performed deformed or needle depth set needle depth if necessary too low Error message during initial Syringe valve not properly...

  • Page 42: Reference Information

    Reference information 6 Reference information 6.1 Description 6.1.1 General A unique sequence of valve switching and syringe dispenser controlled sample withdrawal enables Autosampler A-905 to inject sample in a simple and reproducible manner. Sample volumes down to 1 µl (Autosampler A-905 for ÄKTA) or 0.1 µl (Autosampler A-905 for Ettan) can be injected .
  • Page 43 Reference information 6.1.2 Autosampler A-905 Cover in open position Buffer tubing High-pressure valve Tubing guide Syringe waste Needle arm Syringe dispenser Transport solvent/ wash vial Microplate Wash solvent bottle Wash position Drain wash position Support plate Condensed water and leakage drain Fig.
  • Page 44 Reference information 6.1.3 Rear view COMMUNICATION UniNet 1, UniNet1 computer network Ventilation holes WARNING: MAKE SURE THAT AIR CIRCULATION WILL NOT BE IMPEDED WARNING RISK OF FIRE REPLACE FUSE AS MARKED AVERTISSEMENT RISCUE DE FEU REMPLACEZ FUS COMME INDIQU WARNING RISK OF ELECTRIC SHOCK DISCONNECT POWER BEFORE SERVICING AVERTISSEMENT RISCUE DE CHOC ELECTRIQUE COUPER L’ALIMENTATION AVANT LA MAINTENANCE...
  • Page 45 Reference information 6.1.4 µl pick-up injections The switching sequence for a µl pick-up injection is schematically shown in the following figures. The default position of the injection valve is always INJECT. The injection valve is initially in the INJECT position. The sample needle Pump has entered the vial of transport liquid (mobile phase used to avoid...
  • Page 46 Reference information The programmed injection volume is Pump aspirated from the sample vial. Syringe Column LOAD Needle The sample needle moves back to the Pump transport vial. The sample is quantitatively transported into the loop with transport liquid (mobile Syringe Column LOAD phase) from the transport vial.
  • Page 47 Reference information 6.1.5 Flushed loop injections The switching sequence for a flushed loop injection is schematically shown in the following figures. The default position of the injection valve is always INJECT. The injection valve is initially in the INJECT position. The sample needle Pump and air needle have entered the vial.
  • Page 48 Reference information For flushed loop injections, the sample loop is quantitatively filled by Pump transporting two or more times the loop volume through the loop, Syringe Column LOAD depending on the volume of the loop. Needle The injection valve switches to the INJECT position.
  • Page 49 Reference information 6.1.6 Partial loopfill injections The switching sequence for a partial loopfill injection is schematically shown in the following figures. The default position of the injection valve is always INJECT. The injection valve is initially in the Pump INJECT position. The sample needle and air needle have entered the vial.
  • Page 50 Reference information For partial loopfill injections, the sample loop is filled by transporting Pump the programmed injection volume into the sample loop. Syringe Column LOAD Needle yy y The injection valve switches to the Pump INJECT position. The sample loop is now part of the system mobile phase flow path.

  • Page 51: Technical Specifications

    Reference information 6.2 Technical specifications Sampling Sample capacity 96-well microplate (Low/High) Direct filling or 96 micro vials 384-well microplate Direct filling or 384 micro vials 48-vial adapter 48 vials Maximum total plate height 47 mm (plate + vial + cap/sealing) Minimum total plate height 12.5 mm (plate + vial + cap/sealing) Vial dimensions (cap included)
  • Page 52 Reference information Physical data 280 × 400 × 440 mm Dimensions (W x D x H) Weight 26 kg Electrical data Power supply voltage 100–240 V~, 50–60 Hz Power requirement 250 VA Fuses For 100–120 VAC Two 5.0 AT-fuses (slow, _”x”_, UL/CSA) For 220–240 VAC Two 2.5 AT-fuses (slow, 5 ×...

  • Page 53: Accessories And Consumables

    Reference information 6.3 Accessories and consumables Item Quant./pack Code no. Rheflex ferrule 18-1119-91 Rheflex nut 18-1119-92 Flange tube fitting 1/8" 18-1119-93 Flange tube fitting 1/16" 18-1119-94 Plunger replacement tip 100 µl 18-1119-96 Plunger replacement tip 250 µl 18-1119-98 Sample needle, PEEK, 15 µl 18-1119-99 Sample needle, fused silica, 5 µl 18-1158-68...
  • Page 54 18115442ab_Manual.fm Page 54 Thursday, July 3, 2003 5:42 PM Reference information Item Quant./pack Code no. PEEK tubing, i.d. 0.15 mm, o.d. 1/16" 18-1156-59 PEEK tubing, i.d. 0.25 mm, o.d. 1/16" 18-1120-95 PEEK tubing, i.d. 0.50 mm, o.d. 1/16" 18-1113-68 Tefzel tubing, i.d. 0.25 mm, o.d. 1/16" 18-1121-36 Tefzel tubing, i.d.

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