Amersham Pharmacia Biotech:Spark Holland A-900
Contents
1. Mains switch 230115 V __ Mains input BROT OT 220 240 Vy T Fuses and voltage selector Auto Sampler A 900 User Manual 18 1117 50 Edition AC 23 A Reference information A 3 Flushed loop injections The switching sequence for a flushed loop injection is schematically shown in the following figures The Initial position the injection valve is in the INJECT position The sample needle and air needle have entered the vial The head space pressure applied through the outer air needle ensures that no air or vapour bubbles are Nesib formed during sample aspiration The syringe dispenser aspirates the flush volume from coum the sample vial to fill the sample lines with sample and remove wash solvent Pum lt a 2 1 me pe com The injection valve switches to the LOAD position ae placing a sharp sample front at the inlet of the sample Needle loop a E o For flushed loop injections the sample loop is quantitatively filled by transporting two or more times the eume loop volume through the loop depending on the volume of the loop Auto Sampler A 900 24 User Manual 18 1117 50 Edition AC Reference information A The injection valve switches to the INJECT position The sample loop is now part of the AKTA mobile phase flow path Sample is transported to the colum2. Needle Transport liquid T il M Sample The syringe dispenser aspirates transport liquid from the transport vial to fill the sample line with transport liquid and remove wash solvents Transport liquid The needle moves from the transport vial to the sample vial The injection valve iwitches to the LOAD position Transport liquid Sample The programmed injection volume is aspirated from the sample vial 1 il Transport liquid Sample Auto Sampler A 900 28 User Manual 18 1117 50 Edition AC Reference information A The sample needle moves back to the transport vial The sample is quantitatively transported into the loop with 30 pl of transport liquid sane mobile phase from the transport vial Needle Transport liquid The injection valve switches to the INJECT position The sample loop is now part of the AKTA mobile phase flow path Sample is o transported to the column Transport liquid cc Sample Air segment Normally an air segment shall not be programmed for pl Pick up injections since it then is injected with the sample into the system Needle Injection loop Buffer tubing A Transport Sample Fluent Transport ZAS Note For pl pick up injections there will be no air pressure bea
3. Remove most air bubbles from the syringe Connect the plunger of the filled syringe to the syringe drive and connect the syringe with the Luer connector at the syringe valve Screw the syringe firmly into the Luer connector Remove the air from the syringe by issuing the AutosamplerControl SyringeHome command in Manual Flowpath The syringe will move to its HOME position and dispense its contents to the syringe waste If some air remains in the syringe issue the AutosamplerConirol SyringeEnd command in Manual Flowpath again The syringe is filled with wash solvent Issue the AutosamplerControl SyringeHome command in Manual Flowpath to dispense the wash solvent to waste If there is still air in the syringe repeat the previous action and tap gently on the syringe as the wash solvent is dispensed to the syringe waste Issue the AutosamplerControl InitialWash command in Manual Flowpath to perform a standard wash All tubing connected to the syringe valve will be refilled and flushed The A 900 is now ready for use Auto Sampler A 900 User Manual 18 1117 50 Edition AC Troubleshooting 5 5 Trouble shooting WARNING Always disconnect the power supply before attempting to replace any item on the instrument CAUTION Only spare parts approved or supplied by Amersham Pharmacia Biotech may be used for maintaining and servicing the instrument WARNING The instrument must not be opened by the user It
4. contains high voltage circuits that can deliver a lethal electric shock 5 1 Faults and actions If the suggested actions do not correct the fault call Amersham Pharmacia Biotech Fault Bad reproducibility Injection valve switched dry Syringe valve coupling leaking Little or no peak Peaks of different height Air in the syringe Injection valve switches too slowly Syringe action failed motor generates noise Sample needle bent No injection performed Error meassage during initial wash Auto Sampler A 900 User Manual 18 1117 50 Edition AC Action Replace rotor seal Check and tighten coupling Replace if necessary Syringe leaking problem Replace syringe Ferrule not tightened in injection valve Tighten ferrule Check and tighten coupling Remove the air by issuing the AutosamplerControl SyringeEnd and AutosamplerControl SyringeHome commands in Manual Flowpath If there is still air in the syringe remove it according to section 4 5 Rotor seal worn Replace rotor seal Syringe not tightened enough when replaced Tighten syringe Septa type wrong septa deformed or needle depth set too low Replace needle and readjust needle depth if necessary Syringe valve not properly switched Check syringe 19 5 Troubleshooting Fault Needle destroyed in injection using pl vial segments Ghost peaks at high pressure A 900 does not execute commands No sample injected Ghost or unexpec
5. available on request Adresses Amersham Pharmacia Biotech UK Limited Amersham Place Little Chalfont Buckinghamshire England HP7 9NA Amersham Pharmacia Biotech AB SE 751 84 Uppsala Sweden Amersham Pharmacia Biotech Inc 800 Centennial Avenue PO Box 1327 Piscataway NJ 08855 USA Amersham Pharmacia Biotech AB 1998 All rights reserved Contents Contents 1 ntroduction 111 General ai A 1 EEE o A r a 1 1 3 Safety symbols on the A 900 ainia aaa asis 1 2 Installation 2 1 Unpaid sites ia d din 2 2 2 the A 9 OO sare SUR 3 2 3 POWEr connectionsuu a a EE EESE 3 LA Floid COMNECHON Siles 4 2 59 Preparation of vial vinos l asas 7 3 Operation s l NN 9 4 Maintenance 41 Injection valia oh 11 AD 1Ncedieand tubinesa init Sead desecesd dans tect aes 14 4 3 Syringe sample loop and buffer tubing combinations 15 44 Replacing thesampl needle ninia it 17 AI L 18 5 Trouble shooting S 19 Reference information A Autosampler A 900 MCS CHIP ROI 21 General sci s s n b b 21 A2 struments da 22 Flushed loop 1nj ctiOnss airis aiii AR ay aasAs 24 4 Partial loopf
6. by using the AutosamplerControl RotateTray command in Manual Flowpath Place the tray segment on the front half of the tray CAUTION Do not place or remove a tray segment at the rear half of the A 900 tray This may damage the tray sensors Auto Sampler A 900 User Manual 18 1117 50 Edition AC Operation 3 3 Operation 3 1 General Auto Sampler A 900 User Manual 18 1117 50 Edition AC e Switch on off the A 900 using the mains switch located above the mains inlet on the rear panel The A 900 is operated from UNICORN control system Operating instructions for the A 900 from UNICORN control system is described in a separate User Manual included in the XT upgrade kit for your AKTA chromatography system e When a 500 ul or 1000 ul syringe is used the AutosamplerSyringSpeed command should be set to LOW in UNICORN System Seitings Special e If samples of high viscosity 5 10 cP is used the AutosamplerSyringSpeed command should be set to LOW and the Scaling parameter to 0 2 in UNICOR System Settings Special e Always run the A 900 with the front cover folded down Never open the front cover unless the A 900 is in idle mode 3 Operation 10 Auto Sampler A 900 User Manual 18 1117 50 Edition AC Maintenance 4 4 Maintenance 4 1 Injection valve Description The Injection valve is a two position six port valve The figure shows the flow diagram of the valve The six small circles repre
7. f Mount the new rotor seal The three pins on the shaft assembly fit into the mating holes in the rotor seal Stator locating pin only one way Mount the seal with the grooves facing the stator g Replace the stator ring so that the body locating pin in the stator ring enters the mating hole in the body h Put the new stator face assembly on the stator The Stator support ring three pins on the assembly fit the mating holes in the PEEK version only stator only one way i Mount the stator and stator face assembly on the Stator screws 3 valve so the stator locating pin in the stator ring enters the mating hole in the stator j Replace the three stator screws Tighten each an equal amount until the screws are tight 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 900 for more than one day with buffer solution in the tubings Rinse the A 900 with distilled water prior to longer stops Aqueous buffers or salt solutions Prevent the formation of salt crystals in the valve by flushing out the flow passages and needle port with water after using salt solutions Formation of salt crystals may otherwise damage the injection valve Auto Sampler A 900 User Manual 18 1117 50 Edition AC Maintenance 4 Calibrating sample loops Sample loop sizes are designated by nomin
8. low sample loss and allowing programmable injection volumes pl pick up After aspiration from the vial the sample volume is transported into the loop with transport liquid mobile phase from another vial resulting in no sample loss The A 900 uses a syringe to aspirate the sample from a vial into the sample loop To prevent contamination of the syringe the A 900 is equipped with a buffer tubing between the syringe and the injection valve The wash solvent removes the sample from the buffer tubing and sample needle and also rinses both components 21 A Reference information A 2 Instrument Cover in open position High pressure valve Tubing guide Needle arm Tray segment Syringe waste Syringe dispenser Wash solvent bottle Drain wash position Condensed water and leakage drain Wash position Auto Sampler A 900 22 User Manual 18 1117 50 Edition AC Reference information A O COMMUNICATION _ 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 FUSE COMME INDIQUE o WARNING RISK OF ELECTRIC SHOCK DISCONNECT POWER BEFORE SERVICING AVERTISSEMENT RISCUE DE CHOC ELECTRIQUE COUPER L ALIMENTATION AVANT LA MAINTENANCE o
9. 1 18 1119 90 Tray segment set 0 5 ml vial O D 7 mm 1 18 1120 48 Consumables Rheflex ferrule 10 18 1119 91 Rheflex nut 10 18 1119 92 Flange tube fitting 1 8 5 18 1119 93 Flange tube fitting 1 16 5 18 1119 94 Plunger replacement tip 100 ul 10 18 1119 96 Plunger replacement tip 500 ul 10 18 1119 97 Plunger replacement tip 250 ul 10 18 1119 98 PEEK sample needle 1 18 1119 99 Buffer tubing 2000 ul 1 18 1120 31 Buffer tubing 500 ul 1 18 1120 32 VVash solvent bottle 250 ml 1 18 1120 33 Syringe 100 ul 1 18 1120 34 Syringe 250 ul 1 18 1120 35 Syringe 500 ul 1 18 1120 36 Syringe 1000 ul 1 18 1120 37 Luer lock connection female 1 18 1120 38 PEEK loop 100 ul 1 18 1120 40 PEEK rotor for Rheodyne 9740 valve 1 18 1120 41 Syringe waste tubing 1 18 1120 44 Syringe tubing 1 18 1120 45 Wash vial 1 18 1120 46 Union nut air 1 18 1120 47 Auto Sampler A 900 User Manual 18 1117 50 Edition AC Reference information C Auto Sampler A 900 User Manual 18 1117 50 Edition AC 33 34 C Reference information Auto Sampler A 900 User Manual 18 1117 50 Edition AC November 1998 av eresddn m1 Aq uapams ul p luud Eyesddn qe uoqewsojul 9L amersham pharmacia biotech
10. AKTA design Autosampler A 900 User Manual EE amersham pharmacia biotech mportant user information A Meaning Consult the instruction manual to avoid personal injury or damage to the product or other equipment WARNING The Warning sign is used to call attention to the necessity to follow an instruction in detail to avoid personal injury Be sure not to proceed until the instructions are clearly understood and all stated conditions are met CAUTION The Caution sign is used to call attention to instructions or conditions that shall be followed to avoid damage to the product or other equipment Be sure not to proceed until the instructions are clearly understood and all stated conditions are met Note The Note sign is used to indicate information important for trouble free or optimal use of the product Should you have any comments on this instruction we will be pleased to receive them at Amersham Pharmacia Biotech AB SE 751 84 Uppsala Sweden Trademarks AKTA and UNICORN are trademarks of Amersham Pharmacia Biotech Limited or its subsidiaries Amersham is a trademark of Nycomed Amersham plc Pharmacia and Drop Design are trademarks of Pharmacia amp Upjohn Inc Terms and Conditions of Sale All goods and services are sold subject to the terms and conditions of sale of the company within the Amersham Pharmacia Biotech group which supplies them A copy of these terms and conditions of sale is
11. VAC 50 60 Hz Auto Sampler A 900 User Manual 18 1117 50 Edition AC Reference information B User Manual 18 1117 50 Edition AC Povver requirement Fuses For 100 120 VAC For 220 240 VAC Communication Serial communication port Standards EMC Standards Safety Standards Environment VVorking condition Sound pressure level nstallation category Pollution degree 250 VA Two 5 0 AT fuses slow x _ UL CSA Two 2 5 AT fuses slow 5 x 20 mm IEC 127 UniNet 1 This product meets the requirement of the EMC Directive 89 336 EEC through the harmonized standards EN 50081 1 emission and EN 50082 1 immunity Note The declaration of conformity is valid for the instrument when it is e used in laboratory locations e used in the same state as it was delivered from Amersham Pharmacia Biotech except for alterations described in the user manual e used as stand alone unit or connected to other CE labelled Amersham Pharmacia Biotech instruments or other products as recommended This product meets the requirement of the Low Voltage Directive LVD 73 23 EEC through the harmonized standard EN 61010 1 4 to 40 C 20 95 relative humidity lt 70 dB A Category Il according to IEC 1010 Degree 2 according to IEC 1010 31 C Reference information C Accessories and consumables 32 Item Quantity per pack Code no Accessories Tray segment set 1 5 ml vial O D 12 mm
12. a 1 3 Safety symbols on the A 900 The A 900 contains the following safety symbol SHARP NEEDLE AND MOVING PARTS KEEP HANDS CLEAR This sticker denotes the proximity of the sharp needle and the moving needle arm of the A 900 which if not handled with care could result in injury or damage or destruction of these parts Auto Sampler A 900 User Manual 18 1117 50 Edition AC 2 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 Note Do NOT lift the instrument by the front cover Lift the A 900 with both hands under the instrument or with one hand under the front and the other hand grasping the rear top see figure below Auto Sampler A 900 2 User Manual 18 1117 50 Edition AC Installation 2 2 2 Locating the A 900 The A 900 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 AKTA system base platform Be sure that none of the ventilation holes are blocked Blockage of the ventilation holes can cause malfunctioning of the A 900 Autosampler or even damage the electronics Do not install the A 900 in areas subject to excessive dust direct sunlight or shocks and do not place it near a
13. al values that can vary from the actual value by as much as 20 This variation is due to the tolerance of the tubing ID Since standards and unknowns are usually analyzed with the same loop knowledge of absolute loop volume is rarely needed If the actual loop volume must be known calibrate it in place on the valve so that the flow passages in the valve one in the rotor two in the stator are also taken into account The following PEEK sample loops for the injection valve are available from the valve manufacturer Rheodyne Loop ul Bore I D O D Manufacturers catalog no 6 0 007 1 16 9055 020 10 0 010 1 16 9055 021 20 0 010 1 16 9055 022 50 0 020 1 16 9055 023 100 0 020 1 16 9055 024 200 0 020 1 16 9055 025 500 0 030 1 16 9055 026 1000 0 030 1 16 9055 027 Auto Sampler A 900 User Manual 18 1117 50 Edition AC 13 4 Maintenance 4 2 Needle and tubing The A 900 has separate needles for puncturing and sampling the air needle vvhich 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 follovvs uzunu A Initial position B First the air prepuncturing needle 2 Figure A is lovvered by moving the c
14. ample and remove wash solvent EL Sample The injection valve switches to the LOAD position placing a sharp sample front at the inlet of the sample loop EEL Column For partial loopfill injections the sample loop is filled by transporting the programmed injection volume into the sample loop Auto Sampler A 900 26 User Manual 18 1117 50 Edition AC Reference information A The injection valve switches to the INJECT position The sample loop is now part of the AKTA mobile phase flow path Sample is transported to the column Air segment An air segment should be used to reduce the amount of flush volume This air segment is at the front of the flush volume and will not be injected The air segment can be selected when the injection method is selected in Manual Flowpath Needle Injection loop Buffer tubing A i Sample Eluent Flush Air BL ZA o gt B Sample Eluent Flush mi A Auto Sampler A 900 User Manual 18 1117 50 Edition AC 27 A Reference information A 5 ul Pick up injection The switching sequence for a pl pick up injection is schematically shown in the following figures The Initial poaition the injection valve is in the INJECT position The sample needle has entered the vial of 4 Coum transport liquid mobile phase used to avoid disturbing the chromatogram with an additional peak of the transport solvent
15. d space pressure on the sample vial This avoids sample errors due to air expansion during vial exchange from the sample vial to the transport solvent vial Auto Sampler A 900 User Manual 18 1117 50 Edition AC 29 B Reference information B Technical specifications Sampling Sample capacity Vial dimensions cap included Loop volume Dispenser syringe Vial detection Headspace pressure Switching time injection valve Piercing precision needle Wash solvent Wetted parts Analytical performance Reproducibility Memory effect Physical Dimensions W x D x H Weight Electrical Power supply voltage 30 Standard tray 96 vials of 1 5 ml Std 12 vials per segment Micro vial tray 160 vials of 0 5 ml 20 vials per segment Maximum vial height 47 mm Minimum vial height 32 mm 5 1000 ul 100 ul 250 ul standard 500 ul or 1000 ul syringe Missing vial detection by vial sensor Built in compressor Electrically lt 100 msec 0 6 mm 250 ml internal wash solvent bottle PEEK PTFE glass Tefzel alumina ceramic and inert polymer RSD lt 0 3 for flushed loop injections RSD lt 0 5 for partial loopfill injections injection volumes gt 5 ul 5 ul sample 100ul sample loop lt 2 5 MPa 10 ul sample 20 ul sample loop lt 25 MPa RSD lt 1 0 for ul pick up injections injection volumes gt 5 ul lt 0 1 with programmable needle wash 280 x 540 x 440 mm 22 kg 100 240
16. ect the loose end of needle connection tubing to port 4 of the injection valve use a ferrule made of PEEK e Check the sample needle height The default value for the needle height is 2 mm To check the needle height place a vial in tray segment A1 position 1 Remove tray segment A2 Do an injection and check the needle height in the vial If a tapered vial is used an alternative way of checking the needle height can be useful Do a large injection until air is injected then check the sample level and adjust the needle height e The needle height is adjusted in System Settings Specials Needlehght in UNICORN Wash to clean the new needle by issuing the AutosamplerControl InitialWash command in Manual Flowpath 17 4 Maintenance 4 5 SYRINGE 18 The A 900 is factory equipped with a 250 ul syringe but can also be equipped with a 100 pl 500 pl or 1000 pl syringe To install another syringe proceed as follows Note Do not disconnect the power supply of the A 900 It is needed to move the syringe Note After the syringe change the syringe must be in HOME position Move the syringe to the end position by issuing the AutosamplerControl SyringeEnd command in Manual Flowpath Unscrew the syringe from the syringe valve Make sure that the Luer connector in the valve remains in place Disconnect the plunger from the syringe drive Fill the new syringe with wash solvent
17. ections at the injection valve Waste tubing Y Syringe Waste Condensed water and leakage Drain wash position Auto Sampler A 900 User Manual 18 1117 50 Edition AC General waste Connect the supplied drain tubing to the right hand drain hose connector of the A 900 and put the other end in a bottle placed on the floor This drain removes all the liquid dispensed into the wash position Note Sample that is not injected is also removed through this tubing Syringe waste Place the syringe waste tubing in a small bottle below the A 900 If injection volumes no larger than the volume of the buffer tubing are programmed the syringe waste will only be wash solvent Leakage drain All leakage solvents are drained through the left hand hose connector CAUTION Make sure that the drain and waste tubings are not twisted and obstructing the flow path 2 Installation Wash solvent The A 900 has a built in wash solvent reservoir of 250 ml To position the wash solvent bottle e Fill the wash solvent bottle with the appropriate wash solvent Note Use methanol or a mixture of one part iso propanol in four parts of water as wash solvent Before using the wash solvent degas the solvent with Helium or an ultrasonic bath e Screw the bottle in the wash solvent holder and place the holder on the A 900 Put the wash solvent tubing in the wash solvent bottle e Fill the t
18. er Manual 18 1117 50 Edition AC 15 4 Maintenance 16 e If for some reason an air segment is selected from the InjectionulPickup instruction in Manual Flowpath 5 ul of air is injected in the system together with the sample Injection volumes smaller than 5 ul Partial loopfill Install a 100 pl syringe to obtain maximum reproducibility and accuracy Preferably install a 20 pl 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 This is especially recommended when working with high pressures 15 0 25 0 MPa Note that the minimum sample loss in partial loopfill mode is 30 pl recommended minimum flush volume for the injection For zero sample loss injections use the pl pick injection mode ul Pick up Install a 100 pl syringe for optimum accuracy and reproducibility Note To guarantee quantitative injections do not install a smaller sample loop than 100 pl Injection volumes up to 200 ul With the standard 250 pl syringe standard needle with tubing 15 ul and standard 500 pl buffer tubing but with a 200 pl sample loop the maximum injection volumes are Flushed loop 200 pl sample loss remains 230 nl since loops gt 100 pl need 2 x loop volumes plus 30 pl flushed volume Partial loopfill 100 pl ul pick up 77 ul CAUTION Smaller volumes than 5 pl may be injected but reproducibility and accuracy may not be lt 0 5 for
19. ith two types of tray segments each for vials with a specific diameter see figure Vials used should have the following dimensions Maximum vial height including cap 47 mm Minimum vial height including cap 32 mm Maximum vial diameter Standard vial 12 mm Micro vial 7 mm Auto Sampler A 900 User Manual 18 1117 50 Edition AC 7 2 Installation Filling and sealing the vials The standard vials as well as the conical vials can best be filled by a narrow end pipette to allow air to escape when filling the vial Do not fill vials to the edge If you do sample will be forced into the air needle risking extra cross contamination of samples and fouling of the needle pair It is important that the seal is airtight This maintains a pressure on the vial that prevents air bubbles and to stop evaporation of volatile samples CAUTION Do not re use a sample vial without replacing its cap or septum Note Check the seal after crimping If the cap can be turned easily the seal is not airtight re adjust hand crimper The seal must not be deformed after crimping since this may also deform the needle Loading the sample tray The tray segment types used must be entered in System Settings It is not necessary that all segments are available on the tray at the start of a run To place a segment on the tray there must be an open position in the front half of the tray The tray can be rotated if the A 900 is not running
20. n If one injection is to be made from each vial a wash routine has to be performed after every injection Air segment An air segment should be used to reduce the amount of flush volume This air segment is at the front of the flush volume It will not be injected and will therefore not influence the injection The air segment is selected when the injection method is selected in Manual Flowpath With a standard needle the flush volumes must be minimum 30 pl for injections with an air segment and 35 nl for injections without If the samples are highly viscous it may be necessary to program larger flush volumes and reduce the syringe speed to optimize performance Needle Injection loop Buffer tubing A i i Sample Eluent Flush Air D gt ZZZ 7 ZA E gt B Sample Eluent Flush IZ Z A Auto Sampler 900 User Manual 18 1117 50 Edition AC 25 A Reference information A 4 Partial loopfill injections The switching sequence for a partial loopfill injection is schematically shown in the following figures The Initial position the injection valve is in the INJECT position The sample needle and air needle have entered the vial The head space pressure applied through the outer air needle ensures that no air or vapour bubbles are formed during sample aspiration The syringe dispenser aspirates the flush volume from the sample vial to fill the sample lines with s
21. njection volume 50 of loop volume ul pick up max injection volume loop volume 3 x needle volume 2 standard needle volume is 15 ul Sample volume requirements Partial loopfill iniection volume flush volume using tapered or standard vial ul pick up injection volume using tapered or standard vial Flushed loop loop volume lt 100 ul minimum 3 x loop volume flush volume loop volume 100 499 ul minimum 2 x loop volume flush volume loop volume 500 1000 ul minimum 1 5 x loop volume flush volume The first injection from a vial requires additional sample volume A tapered vial needs 4 ul extra volume and a standard vial needs 300 ul These volumes must be added to the sample volumes given above Note that Flushed loop gives maximum reproducibility but not maximum accuracy vvich depends on the loop used Minimum sample loss 230 pl 2 x loop overfill flush volume for needle Partial loopfill gives maximum accuracy and reproducibility better than 0 5 RSD for injection volumes gt 5 ul Minimum sample loss flush volume 30 pl 30 pl is the recommended minimum flush volume 35 pl is the recommended flush volume with an air segment 40 pl is the default value for flush volume e pl Pick up offers zero sample loss and maximum accuracy same as partial loop fill but slightly less reproducibility RSD is better than 1 for injection volumes gt 5 pl Auto Sampler A 900 Us
22. omplete needle arm dovvnvvards The vial sensor strip 3 Figure A detects if there is a vial and at the same time measures the vial height If no vial is detected the needle arm is moved upwards and the A 900 will stop When the A 900 detects a vial the lovvering speed of the arm is decreased to provide just enough force to puncture the septum of the vial After forcing the air prepuncturing needle into the vial air is applied to the vial An air outlet nut reduces the pressure in the vial to approximately 0 5 bar The pressure in the vial is a function of the diameter of the air outlet nut the air needle and the sample needle C Finally the sample needle 1 Figure A is lowered into the vial to the depth programmed in System Settings Now the A 900 is ready for the first injection from this vial Auto Sampler A 900 14 User Manual 18 1117 50 Edition AC Maintenance 4 4 3 Syringe sample loop and buffer tubing combinations Four sizes of syringes are available for the dispenser 100 250 500 and 1000 ul The 250 pl syringe is the standard syringe When combined with the standard 500 pl buffer tubing and the standard 100 pl sample loop the following injection volume range is available for the various injection modes Flushed loo 100 ul e Partial 1 50 pl ul pick up 1 25 pl The maximum injection volumes are calculated as follows Flushed loop injection volume loop volume Partial loopfill max i
23. partial loopfill or lt 1 for pl pick up For volumes larger than 200 ul Install the 2000 pl buffer tubing Install the appropriate sample loop size and the appropriate syringe Syringe volume gt 2 x injection volume Injection volumes larger than 500 pl are possible but the sample may contaminate the syringe Program sufficient wash after use BUFFER TUBING Syringe Buffer tubing Loop volume 100 ul 500 ul buffer tubing 5 20 ul 250 ul 500 ul buffer tubing 20 200 ul 500 ul 2000 ul buffer tubing 200 500 ul 1000 ul 2000 ul buffer tubing 500 1000 ul Auto Sampler A 900 User Manual 18 1117 50 Edition AC Maintenance 4 4 4 Replacing the sample needle Rheodyne nut and ferrule Teflon sleeve Male nut M6 Needle holder Hex screw Peek sample needle Air outlet nut Air prepuncturing needle Vial sensor strip W Auto Sampler A 900 User Manual 18 1117 50 Edition AC To replace the sample needle proceed as follows e Use the AutosamplerControl NeedleFront command in Manual Flowpath to move the needle to the front the needle connection nut Male nut M6 that fixes the sample needle CAUTION Do not loosen or otherwise unscrew the Hex screw the Rheodyne Rheflex nut that connects the tubing to the injection valve Remove the sample needle by pulling it out of its fitting Put in the new needle assembly Conn
24. sent the ports in the rotor seal Rotating the valve 60 switches Shaft assembly Shaft assembly the valve from one rotated clockwise rotated counterclockwise position to another Specifications e Maximum temperature is 50 C e The valve withstands 34 5 MPa 5000 psi e Wetted surfaces are PEEK alumina ceramic and an inert polymer CAUTION Use only plastic ferrules in the PEEK stator ports Metal ferrules can cause irreparable damage to the plastic stator WARNING Do not operate the A 900 with the front cover open Auto Sampler A 900 User Manual 18 1117 50 Edition AC 11 4 Maintenance Body Shaft assembly Maintenance 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 The procedure for changing the rotor seal stator face assembly and isolation seal is as follows Refer to the figure and proceed a Remove the three stator screws with a 9 64 inch hex key b Remove the stator stator face assembly and Isolation seal stator ring from the valve body c Pull the rotor seal off the pins e Rotor seal d Remove the isolation seal gt e Mount the new isolation seal with the spring side 12 Body locating pin Stator face assembly facing away from the rotor seal Stator ring
25. source of heat 2 3 Power connections Before connecting the mains cord to your AKTA separation unit check that the voltage setting of the A 900 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 reinserting the voltage selector cartridge Check that the right fuses are installed If not replace them with the fuses stated below For 110 120 VAC use two 5 AT fuses slow For 220 240 VAC use two 2 5 AT fuses slow WARNING For continued protection against risk of fire replace only with fuses of specified type and rating See Technical specifications for fuse data When the voltage selection and fuses are correct for your power source plug in the power cord in a free outlet in the base platform of your AKTA separation unit Auto Sampler A 900 User Manual 18 1117 50 Edition AC 3 2 Installation 2 4 Fluid connections WASTE The A 900 is factory installed with a 250 pl syringe a 100 pl PEEK loop a 500 pl buffer tubing and a PEEK sample needle A schematic presentation of all parts that have contact with fluid is shown below To get access to the fluid parts it is necessary to open the front cover of the A 900 therefore remove the screw on the right hand side of the cover The table below lists the dimensions of the standard installed t
26. ted peaks 20 installation Refit if necessary Action e Segments badly oriented or septa bent down Replace needle Bad capillary connections Replace connectors if necessary Syringe in END position Order the syringe to HOME position using the AutosamplerControl SyringeHome command in Manual Flowpath Syringe in END position Order the syringe to HOME position using the AutosamplerControl SyringeHome command in Manual Flowpath All segments in vial tray are not of the same type Change to same type Wrong vial position programmed Program correct position e Washing not performed properly due to Run out of washing liquid VVashing liquid flovvs back in vvashing position beacause vvaste flask not located belovv A 900 Auto Sampler A 900 User Manual 18 1117 50 Edition AC Reference information A Reference information A Description Auto Sampler A 900 User Manual 18 1117 50 Edition AC A 1 General A unique sequence of valve svvitching and syringe dispenser controlled sample withdrawal enables the A 900 to inject sample volumes in a range from a few microliters to 1 ml in a simple and reproducible manner Three different methods for injection can be selected Flushed loop The sample loop is completely quantitatively filled with sample resulting in extremely good reproducibility better than 0 3 Partial loopfill The sample loop is partially filled with sample giving
27. till injections ra R R asias ases 26 AS ul pick up jet miii 28 Technical Specifications triada dida 30 Accessories and consumablesiiinniaai anar 32 User Manual 18 1117 50 Edition AB 1 Contents About this manual This manual comprises two parts a practical part sections 1 5 and a reference part sections A C Sections 1 5 contain information for operating the instrument Autosampler A 900 User Manual 18 1117 50 Edition AC Introduction 1 1 Introduction 1 1 General Autosampler A 900 is an automated sampling injector for use with AKTA explorer and AKTApurifier in liquid chromatography The A 900 features e Sample injection from a few microlitres to 1 ml e Three different methods for injection Biocompatibility e Different sizes of sample vials 1 2 Safety e The instrument is for indoor use only Do not use in a dusty atmosphere or close to spraying water e Do not block the air inlet and outlet of the unit WARNING Do not operate the A 900 with the front cover open WARNING Always disconnect the power supply before attempting to replace any item on the instrument WARNING The instrument must not be opened by the user It contains high voltage circuits that can deliver a lethal electric shock WARNING For continued protection against risk of fire replace only with fuses of specified type and rating See Technical specifications for fuse dat
28. ubing For optimal performance of the A 900 a correct combination of syringe loop and buffer tubing should be installed Loop Pump Dispenser Column Needle WASH BOTTLE INJECT position LOAD position Tubing Material Dimensions Standard sample needle tubing PEEK 135 mm x 0 5 mm OD x 0 25 mm ID Volume 15 ul Buffer tubing from high pressure valve to syringe valve Tefzel 640 mm x 1 16 OD x 1 0 mm ID Volume 500 ul Tubing syringe valve to vvash solvent bottle Tefzel 300 mm x 1 16 OD x 1 0 mm ID Tubing syringe valve to vvaste solvent bottle Tefzel 400 mm x 1 16 OD x 1 6 mm ID 100 pIPEEK loop PEEK 1 16 OD x 0 5 mm ID Tubing general waste Silicon 2x1 2m 10 mm OD x7 mm ID Auto Sampler A 900 User Manual 18 1117 50 Edition AC Installation 2 AKTA connections To ensure reproducible injections make the connections to your AKTA chromatography system according to the installation instructions given in the User Manual included in the XT upgrade kit for your AKTA chromatography system The A 900 has been flushed with 1so propanol make sure that the mobile phase of your AKTA chromatography system is miscible with iso propanol If not start up with an intermediate solvent as mobile phase disconnect the column CAUTION It is essential that the contents of the sample loop are injected in back flush onto the column Therefore do not exchange column and pump conn
29. ubing using the AutosamplerControl SyringeEndand SyringeHomecommands in a Manual Flowpath SyringeEnd aspirates a syringe volume of wash solvent from the wash solvent bottle and fills the wash solvent tubing SyringeHome dispences the syringe contents to the syringe waste Repeat this action until the wash solvent tubing and the syringe are completely filled Syringe waste Wash solvent Inj valve After the wash solvent tubing and the syringe are filled use the AutosamplerControl InitialWash command in Manual Flowpath to perform a standard wash All tubing connected 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 250 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 larger bottle outside the A 900 To fill this wash solvent tubing repeat the previous fill procedure a few times Auto Sampler A 900 6 User Manual 18 1117 50 Edition AC Installation 2 2 5 Preparation of vials Vial dimensions The A 900 is very flexible in the use of vials and septa Almost every type of vial with an OD of 12 or 7 mm can be used from micro vials 0 5 ml to standard vials 1 ml By using inserts the vial volumes of standard vials can be reduced To hold all types of vials the A 900 can be equipped w
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