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User manual AS110, edition 7

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1. Bio compatible sample Inert sample needle coated steel and flow path and valve bio compatible valve PEEK ceramic Air needles 6 different types of air needles are avail able for the AS 110 micro each for a different type of well vial plate However it is not just the type of well vial plate that determines which air needle must be used Refer to the section on Air needles for more information Valve unit Special valve unit that enables easy re placement of valves User manual AS110 edition 7 Injections The autosampler is equipped with a low pressure flow path a compressor a sampling valve and three standard injection modes and an option for user defined programming e Full loop injections for maximum precision e Partial loopfill injections for maximum flexibility e uL Pickup injections for zero sample los e User Defined Program for full flexibility These injection modes accommodate use of a wide variety of applications and are explained in full detail in this chapter Pressure Assisted Sample Aspiration For all injection modes loop injection with Pressure Assisted Sample Aspiration PASA is selectable It is a proven concept that combines high precision with simplicity and reliability e no moving around with the sample needle e reduced risk for bubbles in the sample line e no needle port that wears and contaminates There is only intelligent valve switching and highly accurate syringe buffer tubing
2. control sample loop Air pressure HPLC Pump HPLC Column Figure 9 PASA injection concept Chapter 1 Syringe and buffer tubing The syringe is used to aspirate the sample from a vial into the sample loop Buffer tubing between the syringe and the injection valve prevents contamination of the syringe Wash solvent is used e toremove the sample from the buffer tubing and sample needle e to rinse the buffer tubing and sample needle Three sizes of syringes are available for the AS 110 standard 250 FW 1 26 gt 500 and 2500 uL prep version Using the 500 uL syringe combined with 1000 uL buffer tubing and 100 uL sample loop the following inj volume range is available for the various injection modes e Full loop 100 uL e Partial loopfill 0 50 uL e uL pick up 0 27 uL For the AS 110 micro three sizes of syringes are available 25 uL 50 uL and 100 uL The 25 uL syringe is the standard syringe combined with the 50 uL buffer and the 20uL sample loop the following inj volume range is available for the various injection modes e Full loop 20 uL e Partial loopfill O 20 uL e uL pick up 0 6 4 uL For other combinations of loop syringe and tubing the maximum inj volumes are calculated with the following formulas e Full loop injection volume loop volume e Partial loopfill max inj volume 1 2 x of loop volume e uL Pick up max inj volume loop volume 3 x needle volume
3. AS 110 micro For the 25 uL syringe factory installed the following scale factors apply LOAD L min UNLOAD pL min Scale factor Low Normal High Low Normal High 2 53 80 107 133 160 185 109 218 123 240 137 267 LOAD uL min UNLOAD L min Scale factor Low Normal High Low Normal High Chapter 1 23 For the 100 UL syringe the following scale factors apply LOAD uL min UNLOAD pL min Scale factor Low Normal High Low Normal High 25 50 19 100 125 149 175 100 198 112 223 125 249 AS 110 standard For the 250 uL syringe FW gt 1 26 the following scale factors apply LOAD L min UNLOAD L min Scale factor Low Normal High Low Normal High 137 265 2 3 528 410 800 545 1067 686 1333 813 1600 960 1846 1091 2182 1231 2400 1372 2667 24 User manual AS110 edition 7 For the 500 uL syringe the following scale factors apply LOAD L min UNLOAD pL min Scale factor Low Normal High Low 2 4 545 820 1090 1371 1626 1920 2181 2461 2743 Normal High 930 1055 1600 2133 2666 3200 3691 4363 4800 9333 Chapter 1 25 Mix amp dilute A Mix amp Dilute routine can be created for the AS 110 This routine allows you to process the sample before injection You can program three different types of actions e Add the indicated volume from the Sample Reagent A Reagent B Wash position and dispense it to the Sample Destination position
4. Antec Industrieweg 12 2382 NV Zoeterwoude The Netherlands AS 110 autosampler User manual 191 0010 Edition 7 2015 Ti Q so 9001 T 31 71 5813333 F 3171 5813334 info myantec com www myantec com M certified Copyright 2014 Antec The Netherlands Contents of this publication may not be reproduced in any form or by any means including electronic storage and retrieval or translation into a foreign language without prior agreement and written consent from the copyright of the owner The information contained in this document is subject to change without notice ROXY ALEXYS DECADE DECADE Il INTRO Flexcell ReactorCell SenCell VT 03 u PrepCell SynthesisCell ISAAC HyREF are trademarks of Antec Whatman word and device and Whatrnan word only are trademarks of Whatman International Ltd SOLVENT IFD and AQUEOUS IFD are trademarks of Arbor Technologies Inc Clarity DataApex are trademarks of DataApex Ltd Microsoft and Windows are trademarks of Microsoft Corporation Excel is a registered trademark of the Microsoft Corporation The software and the information provided herein is believed to be reliable Antec shall not be liable for errors contained herein or for incidental or consequential damages in connection with the furnishing performance or use of software or this manual All use of the software shall be entirely at the user s own risk DECLARATION OF CONFORMITY The
5. Doues and Outputs Mic Methods Liar Program Sron Settings tray Long vokse jel Tubang Voie ef 1000 240 Larga Vare p yrange Speed Speed Scat facto Neede Hegri ar l Ae Seperti ELIOT In the Clarity Method window on the AS Systems settings tab set the needle height to the maximum height 6 mm Needle height is measured from the bottom of the sample tray to the needle tip when inserted in a vial Chapter 5 67 Program an injection to be performed from an empty vial that is on the front row of the sample tray and run the injection procedure s soon as the needle is inserted in the vial and reached his lowest point turn off the autosampler and inspect the actual needle height in relation to the bottom of the vial Be quick enough to turn off the autosampler as an actual injection of air should be prevented Turn on the autosampler again Clarity may also need to be restarted after the loss of communication Program a lower needle height inspect the actual needle height and repeat until the desired height is found The optimal value for needle height can now also be programmed adjusted in the user program if applicable 68 User manual AS110 edition 7 Chapter 6 69 CHAPTER 6 Trouble shooting Even though great care was taken in the design of the AS 110 problems may occur e instrument errors these can be caused by a variety of reasons e software errors usually caused by fau
6. In this example three samples are analysed using the full loop injection method Between every injection a needle wash step is performed Method Setup example i x Select Sampler Sampler 1 z I Enabled 45 110 micro autosampler Sampler Method 45 Status Mode Time and Temp Inputs and Outputs Mix Methods User Program System Settings Tray Analysis Time min Injection Mode Flush Volume pl Full Loop 5 00 Wash Volume wl Wash Steps ro o M Tray Thermostatting C o 4 Tray Temperature 4 C Ready AS fc Measurement Acquisition Integration Calculation Advanced Cancel Send method Report Audit Trail Help Status Event Table Figure 25 Injection settings in Mode Time and Temp tab 54 User manual AS110 edition 7 Set the injection mode to full loop injection At the wash pull down selector select wash between injections and set the wash volume to 50 uL In the sample sequence editor 3 injections of 10 uL have to be programmed e Make sure that the sample tray positions A1 to A3 left contains sample vials e Open anew sequence by clicking the sequence icon in the method window e Program three lines with SV 1 to 3 start vial and fill in all other relevant parameters Sample ID sample inj Vol 10 uL method name etc e Click start to execute the sequence 2 Example Sequence Example MODIFIED ioj xi ile Edit Sequence view
7. Integration Calculation Advanced Cancel Send method Report Audit Trail Help Figure 18 AS 110 tray tab Mode Time and Temp Method Setup 100219_1600m xj Select Sampler Sampler 1 7 IV Enabled 45 110 micro autosampler Sampler Method AS Status Mode Time and Temp Inputs and Outputs Mix Methods User Program System Settings Tray Analysis Time min Injection Mode Flush Volume pl 1 0 Ful Loop 15 00 Wash Wash Volume pl Wash Steps between Vials z 50 1 IV Tray Thermostatting C 4 Status Event Table as ic Measurement Acquisition Integration Calculation Advanced Cancel Send method Report Audit Trail Help Figure 19 AS 110 Mode Time and Temp tab The mode Time and Temp tab determines with which injection mode the AS 110 will operate in the method Full loop Partial loopfill u Pickup or User program The other parameters which can be set under this tab Chapter 3 49 e Analysis time this defines the time between two injection cycles when a series of measurements Is programmed in the autosampler e Flush volume is the sample pre flush volume which is used prior to aspiration of sample to fill the sample loop in the full loop and partial loopfill mode e Wash wash between injections or between vials e Wash volume wash volume used for the wash cycle e Tray temperature here the tray cooling temperature can be set
8. To prevent cross contamination the AS 110 will aspirate an additional volume of 25 of the programmed volume to flush the tubing and needle The aspirate and dispense speed depends on the selected syringe and the programmed syringe speed 1 Example ADD 20 uL from Reagent A to Destination will result in the following actions 2 Aspirate an air segment of 0 5 uL to separate the wash solvent in the buffer tubing from Reagent A 3 Aspirate 5 uL Reagent A to flush the tubing and needle 4 Empty the syringe to the syringe waste position 5 Aspirate 20 uL Reagent A and dispense it to the destination vial 6 Rinse buffer tubing and needle with wash solvent e Mix aspirate and dispense a number of the times the programmed volume from the destination vial If no destination vial is available the mix is performed in the sample vial Example Mix 3 times with 25 uL will result in the following actions 1 Aspirate an air segment of 0 5 uL to separate the wash solvent in the buffer tubing from the solvent to be mixed 2 Empty the syringe to the syringe waste position 3 Aspirate 25 uL solvent and dispense it back into the vi al well 4 Repeat step number 3 twice 5 Rinse buffer tubing and needle with wash solvent 26 User manual AS110 edition 7 The mix is performed from the destination position when the previous ADD action is TO DESTINATION When the previous ADD action is TO SAMPLE the mix is performed from
9. User program Under the User program tab a programming interface is available in which fully customized injection methods can be created Dispense steps aspirate steps valve switching IO actions etc can be programmed step by step in an event table for full flexibility With this option advanced mixing programs can be created for automated chemical derivatisation reactions and dilution amp mixing Method Setup INJ userprog 101 e x V Enabled Select Sampler 45 110 micro autosampler Sampler Method AS Status Mode Time and Temp Inputs and Outputs Mix Methods User Program system Settings Tray M Use User Program Injector 2 Reagent A 2 Sample Injector Digital Inject Needle wash 50 00 Status Tray Temperature 4 C Ready AS ic Measurement Acquisition Integration Calculation Advanced Cancel Send method Report Audit Trail Help Event Table Figure 20 User program tab Note for every aspirate dispense step always the syringe speed height has to be defined For more details about the user program steps see chapter 1 paragraph User program 50 User manual AS110 edition 7 Chapter 4 51 CHAPTER 4 Using the AS 110 This chapter describes a number of examples of injection actions that can be performed with the AS 110 using the Clarity software Try to do these examples to learn to work with the AS 110 These examples can be executed
10. B 1E3 However if you were to use a 12 vial tray for the Reagent you could use reagent from the same vial a number of times For example e first line first sample position 2A1 last sample position 2A1 destination position 2E1 Reagent A 1A1 and Reagent B 1E1 e second line first sample position 2A2 last sample position 2A2 destination position 2E2 Reagent A 1A1 and Reagent B 1E1 e third line first sample position 2A3 last sample position 2A3 destination position 2E3 Reagent A 1A1 Reagent B 1E1 etc If you chose to process plates in columns the following positions are available for sample destination reagent A and reagent B OOM OOOOODO sampe MOOOMOOOOH F Destination OOOOOOOC CO Reagenta POOOOOOO reagent OQQOQOOQOOOO OQOQOQOOOOO OQQQOOQOOOO QOQOQOOOOO QOQOOQOOOO OOQQOOQOOOO OOOOOOO m MOOOOOO O OO OOOOO POO OOOO POOOO QOOQOOOO POOOOQOOOO 000000 GOOOO QOQOOOO For example you can now program the following e two 96 low plates processed in columns e first line first sample position 2A1 last sample position 2A1 destination position 2A7 Reagent A 1A1 and Reagent B 1A7 e second line first sample position 2B1 last sample position 2B1 destination position 2B7 Reagent A 1B1 and Reagent B 1B7 e third line first sample position 2C1 last sample position 2C1 destination position 2C7 Reagent A 1C1 Reagent B 1C7 etc The mix method can be executed before the injection
11. after the AS 110 has been successfully installed and after all items described in Chapter 2 and 3 have been correctly set up Example 1 5 uL partial loop fill injection no wash The following example is valid for the AS 110 micro configured with the following items 10 uL sample loop 2 4 uL needle 25 uL syringe 48 positions sample tray Log into the Clarity instrument and open the AS 110 method window and enter the settings as displayed in the next screens xl IV Enabled Select Sampler Sampler 1 AS 110 micro autosampler Sampler Method AS Status Mode Time and Temp Inputs and Outputs Mix Methods User Program System Settings Tray Analysis Time min Injection Mode Flush Volume pl ii Partial Loopfil g Wash Wash Volume yl Wash Steps never z 50 1 IV Tray Thermostatting C 4 Status Tray Temperature 4 C Ready AS ic Measurement Acquisition Integration Calculation Advanced Cancel Send method Report Audit Trail Help Event Table Figure 21 Injection settings in Mode Time and Temp tab In the Mode Time and Temp tab set the injection method to partial loopfill and set a sample flush volume of 5 uL which is about twice the needle volume Set the wash to never and the tray temperature to 4 C Set the analysis time to an appropriate time corresponding to that of your 52 User manual AS110 editio
12. bar for the included 10 port UHPLC valve The following user installable options are available e Bio compatible sample flow path and valve e Prep Kit e Air needles e Valve unit Inert sample needle Silco steel and bio compatible valve PEEK 2 5 mL syringe Prep valve 10 mL sample loop LSV needle and sample tray for 10 mL vials 6 different types of air needles are available for the AS 110 each for a different type of well vial plate However it is not just the type of well vial plate that determines which air needle must be used Refer to the section on Air needles for more information Special valve unit that can be replaced quickly and easily Chapter 1 AS 110 micro The following factory installed option are available for the AS 110 micro autosampler Cooling if installed a cooling fan is visible at the back of the autosampler and a cooling cover is installed inside the sampling compartment Temperature range inside the sampling compartment 4 22 C ISS valve in the sampling compartment This valve can either be used to connect extra solvents or as a column selector SSV valve in the left panel of the autosampler connected to the syringe wash position Allows you to select multiple wash solvents UHPLC version up to 15 000 psi 1000 bar for the included 6 port valve and 10 000 psi 690 bar for the included 10 port UHPLC valve The following user installable options are available
13. be checked Buller tubing LN Pump Detector C fj NOK F ere fl Needle WR Th L Sy Rheodyne valve DY Wash solvent en Syringe waste LL KAAAARKAAAAAKAPLAAKRA AM In the flowcharts on the next pages it is assumed that the AS 110 does not display any error messages Please keep in mind that analytical problems might be caused by external influences like temperature and or light sensitive samples For this reason it is important to make sure that the application was running without problems before and that no changes have been made in the settings System Menu y NO Blockage Disconnect INJECTION in Flow needle from Path Valve Solvent flowing out of inj port Start manual wash Yes No v Disconnect Buffer tubing from valve v Start manual wash Solvent out of open end No y Disconnect Buffer tubing from Syringe valve y Start manual wash Solvent out of syringe valve No y Leakage inside Injection Valve Check for over tightened connections in the entire Flow Path amp Check the syringe valve Disconnect the needle tubing and the buffer tubing Observe ports 3 and 4 for leakage Connect port 1 to an HPLC pump Block port 6 Start the pump at a low flow No Re
14. be programmed on a time base up to 4 times On Off DN Normally closed NC Common COM Normally open NQ Figure 28 Contact closure output Contact closure output Vmax 28 Vdc Vac Imax 0 25 ATTL inputs e Next Injection Input default this input will start the next injection sequence After finishing the injection sequence the AS 110 will wait for the Next Injection Input e Freeze input the AS 110 will freeze the analysis time for the time this input is active If the Freeze Input is activated while the analysis time is not running the AS 110 will perform all programmed pre injection sample handling sample loop But the AS 110 will wait with injecting the sample until the Freeze Input is no longer active e Stop Input with this input the run of the AS 110 is immediately aborted Active high Active l w Autosamel ge re ee Atcsarrpler em f l l YEG 4 En l ai Input Input Lo ei Vr iA ee I Figure 29 TTL input 76 User manual AS110 edition 7 CHAPTER 8 Error Codes The following errors may occur with AS 110 Tray unit ERROR 294 ERROR 295 ERROR 296 ERROR 297 ERROR 298 Needle unit ERROR 303 ERROR 304 ERROR 306 ERROR 307 ERROR 312 ERROR 313 ERROR 315 ERROR 317 ERROR 318 ERROR 319 ERROR 324 ERROR 330 ERROR 331 ERROR 334 ERROR 335 Injection valve ERROR 340 ERROR 341 ERROR 342 Home sensor not reached Deviatio
15. button e Add the AS 110 to the appropriate LC instrument using the gt button in the centre of the System configuration menu e Set the Ext Start Dig Input to device AS 110 micro auto sampler and device 1 in the case you want to use the digital inject trigger of the auto sampler to start a run in Clarity e Another option is to use the analogue start trigger to start a run in that case a detector or A D device should be specified as Ext Start Dig Input to start runs in Clarity For instance the Start input of the DECADE II can be used for this purpose After completing these steps the auto sampler is successfully configured as a part of a Clarity LC instrument and can be accessed via the method menu In case an ALEXYS analyzer or ROXY EC LC system is purchased manual configuration is not required With these products dedicated instrument installers are delivered with pre configured hardware settings Chapter 3 47 AS 110 method menu The Clarity method window contains the following AS 110 settings tabs The most important ones are specified here System Settings Under this tab the following parameters can be set 1 The AS 110 flow path parameters such as loop volume needle volume and syringe volume 2 The injection method parameters such as syringe speed Speed scale factor headspace pressure needle height skip missing vials etc These values affect the standard autosampler injection met
16. its packaging using both hands at the marked position 2 With both hands under the instrument lift the AS 110 to its operating location Keep the instrument upright 34 WARNING CAUTION User manual AS110 edition 7 Position the Alias so that it is possible to access thepower plug Always remove the power cable from the instrument before opening the cover Make sure that the ventilation holes at the back of the autosampler are not blocked Note that if the ventilation holes are blocked this may influence performance and cooling capabilities of the autosampler If objects are placed on top of the AS 110 this may also influence the cooling capabilities Objects can be placed on any side of the AS 110 however make sure that the distance of any objects at the back side of the AS 110 is sufficient to assure optimal ventilation Peltier cooling e 5 cm from the AS 110 if objects are placed at only one side of the AS 110 e 10 cm from the AS 110 if objects are placed on more than one side of the AS 110 Do not place the AS 110 in an area subject to excessive dust or shocks Use the AS 110 indoors only Do not place it near a source of heat or in direct sunlight as this may influence the cooling capabilities of the system 3 Leave the AS 110 to adopt ambient temperature for at least one hour 4 Install AS 110 Service Manager see ASM software on page 45 on your PC 5 Check that fuses and voltage range o
17. method 28 User manual AS110 edition 7 e The injection is performed from the destination position when a mix method is programmed and TO DESTINATION is the last step in the mix method e The injection is performed from the sample position when a mix method is programmed and TO SAMPLE is the last step in the mix method The mix method can also be executed without an injection method Chapter 1 User Program 29 The user program mode is available for the AS 110 micro and for the AS110 standard with FW 1 26 or higher in combination with Clarity software version 4 or higher The User Program offers the possibility to program all possible actions required for a sample handling sequence in separate steps The following User Program steps are available ASPIRATE DISPENSE SYRINGE VALVE SYRINGE NEEDLE WASH a programmed volume from sample well ambient air destination vial wash or one of the reagent vials into the buffer tubing Speed and height of syringe can be entered refer to table below The maximum volume that can be aspirated is the total volume of the syringe a programmed volume from the buffer tubing into the sample well waste destination vial wash or one of the reagent vials Soeed and height of syringe can be entered refer to table below It is not possible to dispense a larger volume than the total volume aspirated in previous actions to program the position of the syringe to one of i
18. reduce the risk of clogging The same applies for the samples make sure you use the appropriate filter material for sample filtration Chapter 3 43 CHAPTER 3 Getting started This chapter describes how to get started with the AS 110 using the DataApex Clarity software The following will be explained e Menus of the AS 110 e Configuration in Clarity e AS 110 method menu e executing a series Menus of the AS 110 Important method service menus in the Clarity AS 110 control module are e Mode Time and Temp here the autosampler analysis time injection mode flush volume wash volume wash type and tray cooling temperature are defined e Input and Outputs In this menu the functionality of the output relay analog inject marker and alarm and inputs next injection Freeze Stop can be defined The IO can be controlled by means of a programmable timed event table e Mix method programming of simple mix amp dilute methods e User program advanced user defined injection programming interface in which fully customized injection methods can be created Dispense aspirate valve switching IO actions can be programmed step by step in an event table e System settings in this tab the AS 110 flow path loop volume needle volume syringe volume etc and injection parameters syringe speed headspace pressure etc are defined for the standard AS 110 injection methods full loop partial loopfill uL pickup e Tray allows you
19. set as the screws close the cap against the valve body Test the valve by pressurizing the system If it does not hold pressure the valve should be returned to Valco for repair 92 User manual AS110 edition 7 AppendixD APPENDIX D Trouble shooting Start up problems oO gt The instrument does not work after switching on of power 93 Power cord Plug in unplugged or Yes replace power faulty cord No Blown fuse Yes Replace fuse No Defective electronics service epresentative Call your r The instrument is working but display doesn t work Display componeni s failure Call your service representative Call your service representative Error code Check Appendix C I appears on the to locate error display message Keyboard componenit s failure No response from keyboard but display works 94 User manual AS110 edition 7 Analytical problems In case of analytical problems you will have to determine whether they are caused by the autosampler or by the rest of the system Quick check e d Replace the valve by a manual injection valve to discriminate between valve problems and other problems Perform a number of manual Flushed loop injections If the results are good the autosampler is faulty if not the HPLC system should
20. to define which trays or well plates are present in the autosampler tray well plate processing direction first destination vial and reagent vial A D positions e Device monitor under the AS 110 micro autosampler tab in the device monitor window important functionality is available for service and maintenance on the autosampler flow path Note that the AS 110 standard and AS 110 micro use a different control module in Clarity The AS 110 standard control module does not have the user program interface available 44 User manual AS110 edition 7 Device monitor The device monitor is an important menu for the maintenance and service of the AS 110 by the end user Some important actions can be performed in this menu such as e The injection valve can be checked switched using Load Inject e The syringe can be moved to its exchange position to be able to remove it for service or replacement Syringe operation 1 e Initial wash to remove air bubbles from the flow path e Set tray cooling temperature e Tray position Home Front e Needle position Home End Exchange Instrument 1 Device Monitor File Control View Window Help ea fil fo ES FASE 27780 lv ROXY Potentiostat Detector 1 d Auto Zero Marker s 35 0 C 1 69 ud 0 035 V I 1 60 Y EJ 45 110 micro autosampler Sampler L ai LO Run Time Temperature I cell Y out E Cell Service l Select Service Operati
21. 0 Syringe dispenser syringe plunger amp plunger tip Replacing the Syringe dispenser valve Fuses Needle height adjustment Trouble shooting Instrument errors Software errors Analytical trouble shooting Vil 33 33 35 37 38 40 40 40 41 43 43 44 47 47 47 48 49 51 51 93 55 55 56 56 57 59 62 64 64 66 66 69 69 70 70 viii User manual AS110 edition 7 I O Connections 74 Error Codes 76 Specifications 79 Accessories amp Spares 83 Calibration amp Performance 87 Maintenance injection valve 90 Disassembly 90 Reassembly 91 Trouble shooting 93 Analytical problems 94 Storage packaging recycling and disposal 97 Decontamination 97 General instructions for disposal 97 Hazardous substances 97 Disposal of electrical components 97 Chapter 1 CHAPTER Introduction 1 The AS 110 standard and AS 110 micro autosamplers are high throughput autosamplers of robust design developed to meet the challenge of the modern analytical laboratory It is a very complete autosampler that needs little bench space the AS 110 is designed for indoor use The AS 110 autosampler features among other things PASA injection concept see page 10 High resolution syringe control this ensures very high precision for injection and reagent addition Internal standard addition sample dilution or derivatization can simply be programmed PC control ensures easy to understand operation context se
22. 0 micro tubing set micro 191 0334U AS 110 micro tubing set 1 32 UHPLC 191 0344 AS 110 syringe valve nut 1 16 micro 191 0346 AS 110 syringe valve flangeless ferrule 1 16 micro 191 0348 AS 110 syringe valve nut 1 8 std 191 0350 AS 110 syringe valve flangeless ferrule 1 8 std 86 User manual AS110 edition 7 Appendix B 87 APPENDIX B Calibration amp Performance The Autosampler is tested on the analytical performance with the following procedure and test configuration PLUMP 1 0 mimin UY LDetectar 7 454 nm Integrator 10 mtr SS tubing 0 25 mm ID for peak modelling Settings Mobile phase and wash solvent H20 Volume of installed loop 100 uL syringe volume 500 uL Syringe speed normal Needle valve tubing volume 15 UL Buffer tubing 1000 uL Tray type i High 96 well Head space pressure no Air segment no Method Test 1 uL pick up Injection volume 10uL Wash Between injections Wash volume 500uL Analysis time 01 00 Sample 50 ppm uracil Injections well 3 First sample Al Last sampler A5 88 User manual AS110 edition 7 Test 2 Partial loopfill Injection volume 10uL Flush volume 45uL Wash Between injections Analysis time 01 00 Sample 50 and 1000ppm uracil Blank Mobile phase H20 Injections well 3 First sample A6 Last sample Fill A1 till A10 with 50 pom Uracil Fill A11 with 1000 ppm Uracil and A12 with H2O Determine for uL pick up the RSD Chrom Perf
23. 0 uL Loop Column Buffer tubing Syringe eS Needle tubing Wash Transport position 5 The injection valve switches to the INJECT position The sample loop is not part of the HPLC mobile phase flow path sample is transported to the column The analysis starts 14 User manual AS110 edition 7 Loop Buffer tubing Column Syringe Qa Needle tubing Wash Transport position A wash routine is performed after each injection Air segment with Full loop injections An air segment of 5 uL 0 5 uL for micro version can be used to minimize mixing of sample during aspiration This air segment is at the front of the flush volume and will not be injected With a standard 15 uL needle the flush volumes must be a minimum of 30 uL air segment 5 uL and 30 uL for injections without air segment With a 2 4 uL micro needle the flush volumes must be a minimum of 4 8 uL air segment 0 5 uL and 4 8 uL for injections without air segment lf samples are highly viscous it may be necessary to program larger flush volumes and reduce the syringe speed for better performance Needle Injection loop Buffer tubing A Sample Fluent Flush Air Eluent O Chapter 1 Injection principles Partial loopfill injection Partial loopfill gives maximum flexibility maximum accuracy plus reproducibility better than 0 5 RSD for injection volumes gt 10 uL for the AS 110 standard For the AS 110 micro the reproducib
24. 0300 191 0302 description AS 110 accessory kit AS 110 micro accessory kit Wash bottle 250 mL PP rectangular Silicone tubing 1 meter AS 110 wash bottle adapter AS 110 wash bottle 100 mL glass I O Cable Tubing connector T piece AS 110 serial cable 9M 9F straight Fuse2A Fuse 2 5 A AS 110 Tray cover shell Spacer 2 5 x 2 5 x 30 AS 110 vial holder 96 low start up kit AS 110 sample vials PP start up kit 48 position vial adapter 12 position vial adapter 84 User manual AS110 edition 7 Needles description 191 0304 Air needle 191 0306 Air needle 80 mm 191 0314 AS 110 sample needle 15 uL 1 16 191 0316 AS 110 sample needle bio 15 uL 1 16 191 0314U AS 110 sample needle 15 uL 1 32 191 0315M AS 110 micro sample needle 2 4 UL 1 16 all all std std UHPLC micro 191 0314M AS 110 micro sample needle bio 2 4 uL 1 16 micro 191 0315U AS 110 micro sample needle 2 4 uL 1 32 191 0332M AS 110 micro needle union 1 32 PEEK Syringes and plunger tips description 191 0338M Sampling syringe 25 uL ILS 181 0342 Sampling syringe 100 uL ILS 181 0344 Sampling syringe 250 uL ILS 191 0326 Sampling syringe 500uL ILS 191 0336M Plunger tip 25 uL pck 5 ILS 181 031 1 Plunger tip 100 uL pck 5 ILS 181 0543 Plunger tip 250uL pck 5 ILS 191 0313 Plunger tip 500uL pck 5 ILS Valves rotor seals and stators description 191 0330 AS 110 syringe valve 181 0324 Valve stainless
25. 110 without the front panel If the cooling option is installed slide out the cooling cover by pulling it gently towards you You can now place well plates User manual AS110 edition 7 RR Figure 6 AS 110 with cooling cover Sample trays The sampling compartment of the AS 110 can accommodate two sampling plates Standard high or low well plates or vial trays 12 48 96 and 384 positions can be used Any combination of well plates is allowed except for 384 Low on the left and 96 High on the right A set of two stainless steel 48 positions sample stays are standard supplied with the AS 110 autosampler Small samples For the sampling of small samples lt 20 uL we recommend to use the following combination of vials and trays Vials caps The vials caps combination that fit in the holder shown in Figure 8 are Sample Vials polypropylene 300 uL Microbiotech pn 4001048 8mm Crimp cap with PTFE seal for single use Chromacol pn 8 ACT NOTE There are subtly different shapes of fraction collector vials on the market The types that fit best in the holder shown in Figure 8 are the ones from Microbiotech depicted on the left side in Figure 7 The types depicted on the right in this figure are slightly too wide to fit well in the 96 position tray but such vials can be sampled by placing them in adaptors that fit in the 48 positions tray Adaptors can be purchased at Antec pn 181 0726 Microdialysis co
26. 2 12 User manual AS110 edition 7 Injection principles Full loop injection Full loop gives maximum possible reproducibility lt 0 8 but not maximum accuracy since the loop volume is specified with an accuracy of 10 The sample loop is completely filled quantitatively with sample This type of injection results in extremely good reproducibility 1 The initial situation the injection valve is in INJECT position The sample needle with air needle has entered the well or vial Headspace pressure applied through the air needle ensures that no air or vapour bubbles are formed during sample aspiration Buffer tubing Syringe lt Needle tubing Wash Transport position 2 The syringe dispenser aspirates the flush volume from the sample well vial to fill the sample line with sample and remove wash solvent Loop Column Buffer tubing Syringe a Needle tubing Wash Transport position 3 The injection valve is switched to LOAD position placing a distinct sample plug at the inlet of the sample loop Chapter 1 13 Loop Buffer tubing Column Syringe Qa Needle tubing Wash Transport position 4 The sample loop is quantitatively filled by transporting a number of times the loop volume through the loop depending on the volume of the loop 3 x loop volume for loop lt 100 uL 2 x loop volume for loops 100 uL 500 uL 1 5 x loop volume for loop gt 50
27. 6 waos o o 7 SwrSpeediHeght Speed 2 Height 3Smm ooo Aspirate 9 00 uL Sample Speed 2 Height 3 5 mm wats SyrSpecdMeigh Speed 2 Peas Aspirate 200 uL Ar Speed 2Height 3B wats BE SyrSpecdHeigh Speed 4 Heh 6Omm EEN Dispense 0 00 uL to Waste Speed 4 Height GOmm TT SyrSpecdHeigh Speed 4 Heght 60mm ooo Sye SpeediHeight Speed 4 Height 6 0 mm ee N z Waiomio Needle wash 300 00 uL Aspirate 10 00 uL Air Speed 4 Height 6 0 mm Ill Move sample to the buffer tubing Dispense 10 00 uL to Waste Speed 4 Height 6 0 mm IV Mixing steps Dispense 10 00 uL to Waste Speed 4 Height 6 0 mm ee 8 a 10 ETEN E 13 mu 15 16 cra 18 19 FOR ai EE 23 24 235 26 28 29 30 Chapter 2 33 CHAPTER 2 Installation Unpacking Inspect the transport box for possible damage as it arrives Immediately inform the transport company in case of damage otherwise she may not accept any responsibility Keep the transport box as it is designed for optimum protection during transport and it may be needed again Carefully unpack the autosampler and inspect it for completeness and for possible damage Contact your supplier in case of damage or if not all marked items on the checklist are included Prior to shipment your autosampler has been thoroughly inspected and tested to meet the highest possible demands Execute the following steps for initial installation of the AS 110 autosampler 1 Lift the AS 110 from
28. Do not disassemble the valve unless system malfunction is definitely isolated to the valve UHPLC valves should not be opened at all and serviced by authorized service personnel of the manufacturer Stator screws gt Pa Figure 30 Exploded view of injection valve Disassembly 1 Use a 9 16 hex driver to remove the socket head screws which secure the cap on the valve 2 To insure that the sealing surface of the cap is not damaged rest it on the outer face Or if the tubing is still connected leave it suspended by the tubing 3 With your fingers or small tool gently pry the rotor away from the driver Appendix C 91 Examine the rotor sealing surface for scratches If scratches are visible to the naked eye the rotor must be replaced If no scratches are visible clean all the parts thoroughly with an appropriate solvent taking care that no surfaces get scratched The most common problem in HPLC is the formation of buffer crystals which are usually water soluble It is not necessary to dry the rotor Reassembly 1 Replace the rotor in the driver making sure that the rotor sealing surface with its engraved flow passages is facing out The pattern is asymmetrical to prevent improper placement Replace the cap Insert the two socket head screws and tighten them gently until both are snug Do not over tighten them the screws simply hold the assembly together and do not affect sealing force which is automatically
29. F CONFORMITY i E Electronic 77 Error Codes 76 User manual AS110 edition 7 F Full loop 12 Fuses 66 G Getting started 43 I O connections 74 Injection valve 56 76 Installation 33 instrument errors 69 L Leaking syringe 71 Leaking syringe valve 71 M maintenance valve 90 Manual 43 Methods programming 43 49 Mix 25 Mix amp dilute 25 N Needle height adjustment 66 Needle unit 76 O open the door 5 Options 8 P Partial loopfill 15 Partial loopfill injections 15 parts 83 Pick up 18 Prep version 8 82 Pressure Assisted Sample Aspiration PASA 10 Index R reference 55 Replacing the sample needle 57 Replacing the Syringe dispenser valve 64 Reproducibility 88 ROHS ii rotor seal 56 Rotor seal worn out 71 RS232 connector 74 S Safety practices iv Sample handling 41 Sample loop 56 Sample positions Mix amp Dilute 26 Sample queue 43 47 ship kit of the AS 110 83 software errors 69 Specifications 79 Specifications Prep version 81 Standard air Needle 59 Symbols iii Syringe and buffer tubing 11 Syringe dispenser 62 Syringe dispenser unit 76 99 Syringe plunger 64 System settings 43 T Tray Cooling 43 Tray unit 76 trouble shooting 93 97 Tubing guide 40 U Unpacking 33 using the AS100 51 V valve maintenance 90 W warning sign iii Wash solvent and syringe rinse 40 Waste t
30. Window Help L fil TAL Z a ES im Sample ID E File Method Name Name 1 Sample 1 Full loopfill i 10 000 Q q No i example Instrument 7 2 Sample 2 Full loopfill 10 000 0 q No example Instrument ia 3 Sample 3 Full loopfill 10 000 Q q No example Instrument P E For Help press F1 Single Run Not ready no method sen Vial Inj File Name Active A Chapter 5 55 CHAPTER 5 Maintenance For all maintenance procedures e Open the door of the AS 110 e lf the cooling option is installed remove the cooling cover by Sliding it towards you e Press the two buttons at the top sides of the AS 110 simultaneously and remove the cover by pulling it towards you You need not disconnect the AS 110 from the power source for any of the maintenance procedures In this way software control will still be possible Use the Direct Control function in the AS 110 Service Manager ASM software to check operation of the various parts of the autosampler Cleaning In general the AS 110 autosampler needs very little maintenance You can clean the outside with a damp cloth with non aggressive cleaning liquid Other items that may need periodic cleaning e valve leak bin see AS 110 autosampler front on page 8 a special leak bin is installed underneath the injection valve You can clean this bin with a damp cloth with non aggressive cleaning liquid e sample tray if sample h
31. are wired Make sure power cords are connected to correct voltage sources Replace blown fuses with the size and rating indicated on the fuse panel and as listed in the list of accessories and spares appendix D of this manual The Alias must only be used with appliances and power sources with proper protective grounding to prevent damage through build up of static electricity Solvents The solvents used may be flammable and toxic The room in which the system is installed should be well ventilated to prevent that solvent vapors Cause poisoning or ignite and cause a fire Use of open fire in the vicinity WARNING Of this system must be strictly prohibited Do not install the system in the CAUTION WARNING WARNING same room with any other equipment that emits or could potentially emit Sparks Provide protective equipment near the instrument solvent gets into the eyes or on the skin it must be flushed away immediately Provide equipment such eye wash stations and safety showers as close to system as possible Sample containers vials should be sealed to minimize any risks related to solvent vapor Do not allow flammable and or toxic solvents to accumulate Follow a regulated approved waste disposal program Never dispose of flammable and toxic solvents through the municipal sewage system Perform periodic leak checks on supply lines Biological Hazard When you analyze biological fluids you need possible precautio
32. ars 2 Click Initialize User manual AS110 edition 7 3 Inthe Initial wash group box click Start to start the wash 4 Click Stop to end the wash 5 Click Close to exit the Direct control window Execute the following steps if you are asked to initialize the system 1 Select AS 110 Direct control The Direct control window appears From this window you can control separate parts of the autosampler to check whether they function as intended 2 Click Initialize to reset the system and prepare it for normal use Execute the following steps if you are asked to switch the system off and then on again 1 Check that the communication cable between AS 110 and PC is properly installed 2 Turn the instrument off with the on off switch at the back of the autosampler 3 Turn the system on again with the on off switch The system is initialized and is now ready for use Software errors Software errors usually are caused by faulty installation of the software or by faulty communication between instruments you will be asked to re install the software on the PC that controls the system lf a software error message appears first check if it may be caused by faulty communication between instruments Check all cable connections between instruments Open AS 110 Service Manager Select the communication port Select AS 110 Direct Control Click Initialize ot IS Analytical trouble shooting Analytical problems like bad rep
33. as been spilled on the sample tray clean the tray with a damp cloth with non aggressive cleaning liquid e drain tubing regularly flush the drain tubing with solvent to prevent clogging and to ensure that liquids and condensate are disposed off 56 User manual AS110 edition 7 Injection valve and rotor seal The AS 110 is equipped with an injection valve either with quick connect mounting or with fixed mounting Execute the following steps to remove the injection valve a 00 OE N N Disconnect all tubing from the valve Only the sample loop can stay in place Remove the valve Remove the screws from the stator part of the valve Gently open the valve and take out the rotor seal Clean and or replace the seal Place the stator back on the rotor and fasten the screws Hold the valve for mounting with port 1 pointing upward Place the valve into its slot and fasten it Reconnect all tubing to the valve In Direct control click Initialize to make sure that the valve is in Inject position 10 Perform a standard wash Direct control Initial wash group box The AS 110 is now ready for use Sample loop The AS 110 is standard fitted with a 10 uL micro or 100 uL sample loop A different sample loop size can be installed but note that you will need the proper combination of syringe and tubing see Syringe and buffer tubing on page 13 to ensure good results Take the following into account when you have instal
34. check with manual valve 95 96 User manual AS110 edition 7 BAD REPRODUCIBILITY Air in Flow Path Start manual wash gt Replace plunger tip or Syringe gt Leaking Syringe Leaking Syringe Valve Replace Valve Replace seal gt Rotor seal worn out Check Stator Redo connections with new Ferrules and nuts Dead volumes in tubing connections AppendixE 97 APPENDIXE Storage packaging recycling and disposal If the autosampler needs to be stored for a long time or if it must be shipped to a different location proceed as follows Thoroughly flush the system Disconnect and remove all tubing except for the sample loop Switch off the autosampler In case of biohazard materials remove and throw away the needle and other flow path materials in accordance with an approved waste disposal program 5 Use the original packaging materials to package the autosampler also place the foam block in the tray location 6 Fill in a health amp safety form for the instrument and include the form in the package 7 Dispatch the package make sure any tax import export requirements are met ee 2M Contact support myantec com if you need more information Decontamination The instrument shall be decontaminated before decommissioning and all local regulations shall be followed with regard to sc
35. d connections to a 6 port valve 38 User manual AS110 edition 7 Bottle Column 2 Pump 2 cI INJECT position Ww LOAD position Figure 12 AS 110 fluid connections to a 10 port valve When making connections to the autosampler sampling valve make sure that the tubing is correctly connected For a 6 port valve e HPLC pump to port 1 of the injection valve e HPLC column to port 6 of the injection valve For a 10 port valve parallel analysis set up e HPLC pump to port 10 of the injection valve e HPLC column to port 1 of the injection valve AS 110 tubing details If you need to install new tubing e insert tube ends always flush with ferrule ends e donot overtighten nuts it may cause blockage in the flow path e make sure that you always use tubing volumes that are suitable for use with the other items in the flow path Chapter 2 39 The AS 110 standard is fitted with the following tubing Tubing Materials Dimensions Standard sample needle and tubing SS 97 mm x 0 8 mm OD x 0 25 mm ID label 15 uL ETFE Tefzel 200 mm x 1 16 OD x 0 25 mm ID Buffer tubing from high pressure ETFE Tefzel 1275 mm x 1 16 OD x 1 0 valve to syringe valve label 1000 mm ID HL Syringe valve to wash solvent bottle PTFE 400 mm x 1 8 OD x 1 6 mm ID Syringe valve to waste PTFE 400 mm x 1 8 OD x 1 6 mm ID The AS 110 micro is fitted with the following tubing Tubing Materials Dimensions Standard sample n
36. e Qa Needle tubing Wash Transport position 6 The sample needle moves back to the transport position A second transport plug is aspirated The sample is quantitatively transported into the loop Loop Column Buffer tubing Syringe Sa Needle tubing Wash Transport position 7 The injection valve switches to INJECT The sample loop is now part of the HPLC mobile phase flow path sample is transported to the column The analysis timer starts Chapter 1 21 Loop Column Buffer tubing Syringe aaa Needle tubing Wash Transport position The sequence is repeated for each injection Air segment with uL Pickup injections If an air segment 5 uL or 0 5 uL for an AS 110 standard and AS 110 micro respectively has been programmed it appears at the front of the first plug of transport liquid and at the front of every sample plug In this injection mode e the air segment at the front of the sample plug is injected into the HPLC system e no headspace pressure can be applied on vials wells in this mode to avoid sample errors due to air expansion during exchange from the sample vial well to the transport position Needle Injection loop Buffer tubino nsport Sample Air Transport Eluent 22 User manual AS110 edition 7 syringe speed scale factor The standard injection procedures use a programmable syringe speed The tables below show the flow rates for each code
37. e max 9 hr 59 min 59 sec Wash Programmable Wash between injections and Wash between vials Timed events Programmable 4 x AUX ON OFF Priority sample Programmable Communication 1 programmable relay output programmable as Inject marker default Auxiliary Alarm 2 programmable TTL inputs programmable as Next injection input default Freeze input Stop input Serial communication port RS232C Options factory installed Sample tray cooling Built in Peltier cooling Range 4 C to Ambient 3 C Temp air temperature in sample compartment 4 C 2 at temperature sensor Temperature at relative humidity of 80 and ambient temperature of 25 C Options user installable Bio compatible sample flow Inert sample needle Silco steel and bio path and valve compatible valve PEEK Prep Kit See Specifications 2 5 mL syringe Prep valve 10 mL sample Prep version on page 74 loop LSV needle and sample tray for 10 mL vials User manual AS110 edition 7 PREP VERSION Note that this specification only lists items that are different from the standard AS 110 specification The Prep version of AS 110 is designed for Large Sample Volumes LSV Sampling Sampling capacity 24 vials of 10 mL LSV Vial dimensions cap included Maximum vial height 47 mm Minimum vial height 32 mm Loop volume Not programmable injection volume determines the aspirated sample volume Dispenser syringe 2500 uL syringe Injec
38. ect and determine the RSD and carry over of the partial loopfill injection by RSD l x100 Carry over poe oo Peak area Peak area Sl ppm The calculated RSD and carry over must be within the following specs Reproducibility RSD lt 1 0 for ul pick up injections RSD lt 0 5 for partial loopfill injections Carry over lt 0 05 with programmable needle wash N B The final test is programmed in AS 110 Service Manager ASM Via service and validation life test there can be chosen between 4 experiments under validation test Those are 1 uL pickup 2 partial loopfill 3 full loop 4 prep mode By choosing the experiment and clicking on start the final test will be automatically processed Appendix B 89 90 User manual AS110 edition 7 APPENDIX C Maintenance injection valve The AS 110 is standard equipped with a Valco C2 2006 injection valve The AS 110 micro with either a Valco C2 1006 or C2 1000 injection valve The AS 110 micro cool UHPLC is equipped with a Valco C72NX 6676 injection valve Cleaning a valve can often be accomplished by flushing all lines with appropriate solvents With normal use the valve will give many tens of thousands of cycles without trouble 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 which can scratch the rotor seal Following is the procedure for changing the rotor seal NOTE
39. ect buffer tubing from valve Start a manual wash If solvent flows from open end check rotor seal if not disconnect buffer tubing from syringe valve Start a manual wash If solvent flows from syringe valve check buffer tubing if not check for over tightened connections in the entire flow path and check the syringe valve CAUTION 73 Clogging of the needle The internal diameter of the AS 110 micro and flow path sample needle is very small and will easily get clogged Filtering the eluent with 0 2 um filter will considerably reduce the risk of clogging The same applies for the samples make sure you use the appropriate filter material for sample filtration Leakage in the Disconnect the needle tubing and injection valve buffer tubing Connect port 1 to an HPLC pump Block port 6 Start the pump at a low flow Observe ports 3 and 4 for leakage If leakage occurs at ports 3 and 4 check rotor seal if not recheck with manual valve Observe the maximum allowed pressure of 350 bar to prevent leakage in the valve In case of an UHPLC version of the AS 110 the maximum allowable pressure is 680 bar for the 6 port valve and 1030 bar for the 10 port valve 74 User manual AS110 edition 7 CHAPTER 7 I O Connections WARNING The AS 110 has two I O connections e RS232 connector for serial communication using the SparkLink protocol e Contact closures output and TTL inputs connector The manufacturer will
40. ed after August 13 2005 are labelled with the crossed out wheelie The symbol on the product indicates that the product must not be disposed as unsorted municipality waste When taking the instrument out of service the different materials must be separated and recycled according to national and local environmental regulations Collection amp recycling information business to business Antec Leyden offers the possibility for disposal and recycling of their instrument at an appropriate recycling facility if requested there may be costs involved with this service Please contact Antec for more information about this service and to register the return and disposal of end of life instruments info myantec com To assure hygienic amp personal safety all instrument should be returned with a signed decontamination form which is available on the website Shipping address for end of life products Antec Industrieweg 12 2382NV Zoeterwoude The Netherlands In case of questions or if further information is required about the collection amp recycling procedure please contact Antec or your local distributor ROHS directive The AS 110 is ROHS compliant and in conformity with Directive 2011 65 EC Restricted use of Hazardous Substances in electrical and electronic Equipment ROHS Antec Leyden is an ISO 9001 2008 certified company About this manual This guide is written for laboratory technicians who use the AS 110 au
41. ee area around instrument Weight Max weight that can be placed on top of AS 110 Power requirements Viscosity range Sampling Sample capacity Vial Plate dimensions incl Cap Loop volume 19 LAeq lt 70 dB 10 40 C indoor use only 25 60 C 20 80 RH According to EC directives cCSAus CSA UL approved II 2 up to 2000 m 300 mm x 510 mm x 360 mm without cooling option 300 mm x 575 mm x 360 mm with cooling option Minimum free distance of 5 cm from obstacles at rear side and air outlets of the cooling units 19 kg without cooling 21 kg with cooling 65 kg 95 240 Volt AC 10 50 60 Hz 200VA 0 1 5cP 2 Micro Titre Plates according to SBS standards 96 well high low and 384 well low formats 48 vial or 12 vial trays any combination of plates is allowed except for 384 Low left and 96 High right Max plate vial height 47 mm incl septa or Capmat Standard 1 5000 UL programmable with 1 uL 10 mL loop optional Micro 0 10 20 00 uL programmable with 0 01uL increments 80 Dispenser syringe Vial detection Headspace pressure Switching time injection valve Piercing precision needle Wash solvent Wetted parts in flow path Injection cycle time Analytical performance Injection modes Reproducibility AS 110 500 uL syringe Reproducibility AS 110 micro 25uL syringe Programming Interface Injection methods Injection volume User
42. eedle and tubing SS 136mm x 0 8 mm OD x 0 1 mm ID label 2 4 uL PEEKsil 165 mm x 1 32 OD x 0 1 mm ID Buffer tubing from high pressure PEEK 260 mm x 1 16 OD x 0 5 mm ID valve to syringe valve label 50 uL Syringe valve to wash solvent bottle PTFE 400 mm x 1 16 OD x 0 75 mm ID Syringe valve to waste PTFE 400 mm x 1 8 OD x 1 6 mm ID The AS 110 micro is fitted with the following tubing Tubing Materials Dimensions Standard sample needle and tubing SS 136mm x 0 8mm OD x 0 1 mm ID label 2 4 uL PEEKsil 165 mm x 1 32 OD x 0 1 mm ID Buffer tubing from high pressure PEEK 260 mm x 1 82 OD x 0 5 mm ID valve to syringe valve label 50 uL Syringe valve to wash solvent bottle PTFE 400 mm x 1 16 OD x 0 75 mm ID Syringe valve to waste PTFE 400 mm x 1 8 OD x 1 6 mm ID 40 User manual AS110 edition 7 Tubing guide To prevent that the wash tubing obstructs the horizontal movement of the needle unit use the tubing guide integrated in the leakage drain Figure 13 Tubing guide I and top view of tubing guide r Waste tubing Make the following connections for disposal of waste liquids e General waste connect the drain tubing in the ship kit of the AS 110 to the right hand drain hose connector see Figure 2 number 2 Place the other end in a bottle for waste on the floor Through this drain all the liquid dispensed to the wash position is removed Sample liquid that is not injected i
43. errules and tubing connections nuts Carry over Causes Solutions Bad match between Check hardware sample characteristics Needle either use an extra wash to and hardware wash the inside and outside needle or install a different type of needle Steel or Silica coated Valve replace rotor in valve Tubing install different tubing Steel Peek between autosampler and column or use different wash solvents Sticky sample Change wash solvents or use multiple wash solvents 72 User manual AS110 edition 7 If a blank gives a peak that is too high for your criteria Causes Solutions Solubility problem You can either modify your sample or accept Carry over Bad match between Check hardware sample Needle either use an extra wash to wash characteristics and the inside and outside needle or install a hardware different type of needle Steel or Silica coated Valve replace rotor in valve by Valco E or H type Tubing install different tubing Steel Peek between autosampler and column or use different wash solvents The blank you use Use a new blank has been soiled Cause not clear Check if you can solve the problem by using more variation in solvents If no injection takes place Causes Solutions Blockage in flow path Disconnect needle from valve Start a manual wash If solvent flows from the injection port check the needle if no solvent flows from the injection port disconn
44. for a 25 uL syringe speed 5 for a 50 uL syringe and speed 4 for a 100 uL syringe If more viscous liquids are used the speeds should be reduced CAUTION Example The following example is valid for the AS 110 micro configured with the listed items below 1 5 uL sample loop 2 4 uL needle 25 uL syringe The example of a user defined program Table 2 will perform an in needle derivatisation procedure e g for GABA Glu analysis followed by an injection according to the schematic principle depicted in Figure 10 User manual AS110 edition 7 Schematics of fluids In sample tubing ao 3 I Aspirate air plug for insulation from wash solution Vv 1 Aspirate Air 2 uL O gt ll Aspirate sample and reagent 2 Aspirate Reagent A 0 5 uL 3 Aspirate Sample 9uL Lo lll Move sample to the buffer tubing i i C D 4 Aspirate Air 12 uL IV Mixing 5 Dispense Waste 10 uL az 6 Aspirate Air 10 uL la 7 Dispense Waste 10 uL E E q V Inject and wash 8 Injector valve LOAD Sample 9 Dispense Waste 7 uL loop 10 Injector valve INJECT g 11 Needle wash 250 uL r Figure 10 Principle of automated in needle derivatization procedure Table 2 User defined program for in needle derivatisation procedure SwrSpeedtieghSesed a ess OOOO TT 2 Asprate 200 uL Ar Speed 4 Height 35mm Aspirate air plug for neuaton T 4 SyrSpeedHeight Speed i Height 60mm 5 Aspiale 050 ul Reagent A Speed 1 Height 50mm N Aspirate reagi
45. he syringe remove them by gently tapping the syringe 14 Connect your HPLC pump to port 1 of the injection valve and the column or the capillary to port 6 of the injection valve Check for leakage and let the system stabilize for at least 5 minutes ASM software A software package is supplied with the AS 110 autosampler AS 110 service manager ASM AS 110 Service Manager allows you to upload AS 110 firmware It offers the following functionality e You can define a port through which you want to do the update e You can access the Communication Settings Direct control Service and Adjustment windows for AS 110 36 User manual AS110 edition 7 e You can access ASM help and About Execute the following steps to install ASM 1 Check the Antec website www myAntec com for the latest version and store it to a convenient location or insert the CD into the CD drive of your PC 2 Ifthe autorun feature is active the installation wizard will appear If autorun is not active use the browser to go to the CD drive and double click install exe to start the installation wizard 3 Answer all questions that pop up in the wizard click Next to go to the next step in the installation procedure 4 Click Finish to end the installation procedure 5 The software is now installed Refer to online help of ASM for more information Upload notification If the upload notification is displayed the ASM has discovered a problem in PC settings tha
46. he syringe valve in the an optional port position before you replace the syringe valve In this position the mounting screws are opposite in line with the holes 1 Move the syringe to exchange position use the Exchange button in Direct control 2 Loosen the lower socket head screw number 2 a full rotation counter clockwise 3 Loosen the upper socket head screw number 1 a full rotation counter clockwise 4 Remove the syringe 5 Remove the syringe valve and install a new one Make sure the flat side of the axle faces forward and make sure that the valve is completely pushed upward 6 Install the new seal 7 Fasten the two socket head screws again fingertight 1 4 turn S Eib eee ee meee ee See Figure 27 Replacing the syringe dispenser valve 66 Fuses User manual AS110 edition 7 Fuses of the following types are installed in the AS 110 2 X 2 5 Disconnect the AS 110 from its power source if you need to replace fuses ELECTRICAL SHOCK If you need to replace the fuses make sure that you install fuses of the same type and rating Fuses are in the fuse box at the back of the autosampler Contact Service if problems with fuses are recurring Needle height adjustment Program a method in Clarity with partial loop fill or full loop fill method SSS en xj Seiet Senge gt 25 D0 mon Sanel Method Mode Tine and Teme
47. hods only full loop partial loopfill uL pickup These settings will not apply for the user program Method Setup example i xj V Enabled Select Sampler Sampler 1 45 110 micro autosampler Sampler Method AS Status Mode Time and Temp Inputs and Outputs Mix Methods User Program System Settings Tray Loop Volume pl Tubing Volume pl 10 2 40 Syringe Volume pl Syringe Speed Speed Scale Factor Needle Height mm 25 Low v 0 5 5 0 IV Air Segment IV Reset Output Skip Missing Vials JV Headspace Pressure Status Not Ready No loaded method AS uc Measurement Acquisition Integration Calculation Advanced i i Cancel Send method Report Audit Trail Help Event Table Figure 17 AS 110 system settings tab Tray The tray tab allows you to define which trays or well plates are present in the autosampler tray well plate processing direction first destination vial and reagent vial A D positions 48 User manual AS110 edition 7 Method Setup example x Select Sampler Sampler 1 V Enabled AS 110 micro autosampler Sampler Method AS Status Mode Time and Temp Inputs and Outputs Mix Methods User Program System Settings Tray Left Tray Right Tray Plate Processing Rows I Reagent A I Reagent B l Reagent C I Reagent D Status Not Ready No loaded method Event Table as uc Measurement Acquisition
48. ility is better 1 0 RSD for injection volumes gt gt 0 5 uL Minimum sample required Flush volume 30 uL AS 110 std or 4 8 uL AS 110 micro These are the recommended minimum flush volume smaller flush volumes can be programmed but will result in decreasing performance The switching sequence for a partial loopfill injection is The initial situation the injection valve is in the INJECT position The sample needle with air needle has entered the vial well Headspace pressure applied through the outer air needle ensures that no air or vapour bubbles are formed during sample aspiration Loop Column Buffer tubing Syringe Qa Needle tubing Wash Transport position The syringe dispenser aspirates the flush volume from the sample vial to fill the sample line with sample and remove wash solvent 16 User manual AS110 edition 7 Loop Column Syringe Qa Needle tubing Wash Transport position The injection valve switches to LOAD placing a distinct sample plug at the beginning of the sample loop Loop Column Buffer tubing Syringe Sa Needle tubing Wash Transport position The programmed injection volume is now aspirated into the sample loop Loop Buffer tubing Column Syringe lt Needle tubing Wash Transport position Chapter 1 17 5 The injection valve switches to INJECT The sample loop is now part of the HPLC mobile phase f
49. illed for more than 60 the air needle can be applied as usual The same applies for the deep wells If you need to deviate from these standard settings use one of the optional needle types Which air needle for which titre plate or vial To determine which air needle to use the following dimensions need to be considered e the height of the titre plate in mm Ht FO e well depth in mm Dw e thickness of capmat or seal in mm Cd e set needle height in mm Nh e distance air needle point through the capmat or seal in mm min 2 mm Ac The following must be true Ay Ht Dw must be between 2 and 6 mm If this is true the protrusion length of the sample needle can be calculated this is the distance between the point of the sample needle and the point of the air needle It can be calculated as as follows 1 Protrusion length Ht Cd Nh Ac Protrusion length Air needle type from to 50 mm yellow 56 mm red 62 mm white std needle 68 mm blue 74mm green 80 mm black Chapter 5 61 You can select the most suitable air needle on the basis of the protrusion length 10 mL vial 50 mm air needle 2 mL vial 62 mm air needle Greiner deep well M53000 Greiner low well 80 mm air needle with capmat 56 mm air needle Example You have a Greiner deep well with Micronic capmat M53000 the AS 110 has a standard need
50. le height setting Calculations will be as follows Ht 41 4 mm The following is true Dw 37 8 mm 41 4 37 8 3 6 is between 2 and 6 mm Cd 3 8 mm Protrusion length 41 4 3 8 6 0 2 0 29 6 Nh 6 0 mm standard Ac 2 0 mm minimum An air needle of 56 mm is required 62 User manual AS110 edition 7 Air needle replacement Execute the following steps to replace the air needle 1 Remove see Replacing the sample needle on page 29 the sample needle Unscrew the chrome locking nut to remove the air needle Unscrew the chrome locking nut from the adjustment nut Get the new air needle Screw the height adjustment nut to the chrome locking nut thread of the height adjustment nut must be level with the lower part of the locking nut Make sure the O ring seal is in the locking nut Install the air needle Install the sample needle 8 Program the proper needle height for the new needle in the ASM settings window Go to Adjustments to adapt Needle Tray settings if necessary oa ee ND If you use trays with 12 vials or 48 vials make sure that the needle height settings is gt 2mm to prevent the needle from touching the bottom of the vials CAUTION 9 Doan initial wash from Direct control to rinse the needle Syringe dispenser The AS 110 is standard supplied with a 500 uL syringe but a 2500 uL syringe can also be installed for the Prep version The AS 100 micro is standard supplied
51. led a sample loop connect the loop between ports 2 and 5 of the injection valve go to the configuration settings and adapt settings in the flow path group box if you have installed a loop with a different volume Chapter 5 57 Replacing the sample needle Take great care when replacing the 2 4 uL micro sample needle tubing to the needle has a very small internal diameter and over tightening may result in blocked tubing Execute the following steps to replace the sample needle 1 Open Direct control AS 110 Service Manager 2 Click Exchange in the Needle group box The needle moves to exchange position 3 Loosen the needle connection nut number 3 58 User manual AS110 edition 7 1 Turn the needle clockwise to loosen it from its bracket number 4 2 Loosen the nut number 1 that connects the tubing number 2 to port 4 of the injection valve 3 Remove the sample needle by pulling it out of its fitting by the tubing 4 Install anew needle assembly 5 Bend the needle tube towards you and turn the needle counter clockwise to hook it into the needle bracket number 4 6 Tighten the needle assembly with the needle connection nut 7 Connect the other end of the needle connection tubing to port 4 of the injection valve Do not tighten too much as this may block the tubing 8 Click Initialize in Direct control The sample needle moves back to home position 9 Perform a wash routine to clean the new
52. ll but slightly lower reproducibility AS 110 standard RSD better than 1 for injection volumes gt 10 uL AS 110 micro RSD better than 2 5 for injection volumes gt 0 5 uL The switching sequence for UL pickup injections is 1 In the initial situation the injection valve is in INJECT position The sample needle has entered the transport position Buffer tubing Syringe Ss Needle tubing Wash Transport position 2 The transport reservoir is filled with wash solvent the programmed amount of times the syringe volume after a wash or after emptying of buffer tubing The injector valve remains in INJECT position during fill transport Please note that the transport solvent needs to be compatible with eluent Column Buffer tubing Syringe Bay Needle tubing Wash Transport position Chapter 1 19 3 For the first injection the syringe dispenser aspirates a transport plug from the transport position to fill the sample line with transport liquid and remove wash solvents Loop Buffer tubing Syringe eee Needle tubing Wash Transport position 4 The needle moves from the transport position to the sample vial The injection valve switches to LOAD position Loop Buffer tubing Syringe Needle tubing Wash Transport position 5 The programmed injection volume is aspirated from the sample vial 20 User manual AS110 edition 7 Column Buffer tubing Syring
53. ll vial adapters 100pcs Chapter 1 Figure 7 Picture of two slightly different fraction collector vials Tray Form a stabile holder for narrow fraction collector vials by inserting the following two plates Figure 8 into each other transparent 96 positions plate Greiner bio one pn 652280 96 positions flat bottom plate Greiner bio one pn 655101 A set for four plates for one autosampler can also be ordered at Antec AS 110 vial holder 96 low start up kit p n 191 0600 Desesered teter 144 p oer vig petet y perre Bal reed Hd FYN 4 ns OSes Figure 8 Left transparent 96 positions plate and right 96 positions flat bottom PCR plate The two plates can form a stabile holder for narrow fraction collector vials lower picture showing 5 inserted vials Configuration options AS 110 standard User manual AS110 edition 7 The following factory installed options are available for the AS 110 autosampler e Cooling if installed a cooling fan is visible at the back of the autosampler and a cooling cover is installed inside the sampling compartment e Prep version AS 110 suitable for large volume sampling Because larger volumes must be injected for the Prep mode see Specifications Prep version on page 52 AS 110 is fitted with a 2500 uL syringe and a 10000 uL sample loop e UHPLC version up to 15 000 psi 1000 bar for the included 6 port valve and 10 000 psi 690
54. low path sample is transported to the column The analysis starts Loop Column Buffer tubing Syringe Ga Needle tubing Wash Transport position If an injection from the same vial and no wash routine is programmed the next injection sequence will start with a flush of 50 of the programmed flush volume Otherwise it will start with a flush of the programmed flush volume If the withdrawal of sample for the next injection exceeds the total volume of the sample buffer tubing the buffer tubing is rinsed before the next injection The next injection will start with the programmed flush Air segment with Partial loopfill injections An air segment of 5 uL 0 5 uL for micro version can be used to minimize mixing of sample during aspiration This air segment is at the front of the flush volume and will not be injected With a standard 15 uL needle the flush volumes must be a minimum of 30 uL air segment 5 uL and 30 uL for injections without air segment With a 2 4 uL micro needle the flush volumes must be a minimum of 4 8 uL air segment 0 5 uL and 4 8 uL for injections without air segment If samples are highly viscous it may be necessary to program larger flush volumes and reduce the syringe speed for better performance Eluent Flush Air Eluent fff 18 User manual AS110 edition 7 Injection principles UL Pickup injection Pick up offers no sample loss maximum accuracy Same as partial loopfi
55. ls lil Safety instructions iV Working environment amp safety IV System Operation iV Electrical safety IV Solvents iV Biological Hazard V Waste disposal V Applications quality control V Introduction 1 Autosampler configurations 2 Instrument description 3 AS 110 autosampler front 3 AS 110 autosampler back 4 Access 5 Sample trays 6 Configuration options 8 AS 110 standard 8 AS 110 micro 9 Injections 10 Pressure Assisted Sample Aspiration 10 Syringe and buffer tubing 11 Injection principles Full loop injection 12 Air segment with Full loop injections 14 Injection principles Partial loopfill injection 15 Air segment with Partial loopfill injections 17 Injection principles uL Pickup injection 18 Air segment with uL Pickup injections 21 Syringe speed scale factor 22 AS 110 micro 22 AS 110 standard 23 Mix amp dilute 25 Sample positions Mix amp Dilute 26 User Program 29 Example 31 Installation Unpacking ASM software AS 110 flow path AS 110 tubing details Tubing guide Waste tubing Wash solvent and syringe rinse Sample handling Getting started Menus of the AS 110 Device monitor AS 110 method menu system Settings Tray Mode Time and Temp User program Using the AS 110 Example 1 5 uL partial loop fill injection no wash Example 2 three full loop injections 10 uL including wash Maintenance Cleaning Injection valve and rotor seal Sample loop Replacing the sample needle Air needles for AS 11
56. lty communication between instruments or by faulty installation of the software e analytical problems see Analytical trouble shooting these may occur e g as a result of wear of parts errors in injection settings and methods or a wrong combination of sample loop buffer tubing and syringe AS 110 Service Manager contains a Service option select AS 110 Service Note that an access code is required for this option and that the service option is intended for service engineers only Contact your supplier if a problem occurs that you cannot solve Instrument errors Incidental fault conditions may occur in any instrument The AS 110 will generate an instrument error message with an error number a short description of the error and instructions on how to proceed In most cases you will be asked to either initialize the system or to switch the system off and then on again Always click OK and follow the instructions to resolve the error status Use AS 110 Direct control in AS 110 Service Manager to monitor the error Initialize the system in the AS 110 Direct control window Make sure the AS 110 is connected to a grounded power source If the LED is not lighted a fuse may have blown WARNING Checking a valve implies that you remove the valve and check all parts for wear and dirt Execute the following steps after any problem with a valve has been resolved 1 Select AS 110 Direct control The Direct control window appe
57. manual AS110 edition 7 Standard 500 uL standard or 2500 uL for Prep option Micro 25 uL standard or 50 uL and 100 uL optional Missing vial well plate detection by sensor Built in compressor but only for vials with septa Electrically lt 100 msec 0 6mm Integrated wash solvent bottle SS316 PTFE TEFZEL VESPEL Glass Teflon Optional PEEK lt 60 sec in all injection modes for 1 injection lt 100 uL including 300 uL wash Full loop partial loopfill and uL pickup PASA pressure assisted sample aspiration RSD lt 0 3 for full loop injections RSD lt 0 5 for partial loopfill injections injection volumes gt 10 uL RSD lt 1 0 for uL pickup injections injection volumes gt 10 uL valid at 1 0 cP RSD lt 0 3 for full loop injections RSD lt 1 0 for partial loopfill injections injection volumes gt 0 5 UL RSD lt 2 5 for uL pickup injections injection volumes gt 0 5 UL lt 0 05 with 100 uL needle wash AS 110 Service Manager software DataApex Clarity control module Full loop partial loopfill and UL pickup Standard 1 uL 5000 uL with 1 uL increment depending on system settings Micro 0 10 uL 20 00 uL with 0 01 uL increment depending on system settings Chapter 9 81 Max injection volume Full loop loop volume Partial loopfill 1 2 x of loop volume uL Pick up loop volume 3 x needle volume 2 Injections per vial well max 9 injections Analysis tim
58. manufacturer hereby declares that the product AS 110 auto sampler type 191 To which this declaration relates is in conformity with the following directives Machinery directive 2006 42 EC Low voltage directive 2006 95 EC applied with the following standards EN 61010 1 2010 EN 61010 2 081 2001 A1 Equipment Class I Installation cat Il Pollution degree 2 EMC directive 2004 108 EC applied with the following standards EN 61326 1 2006 ROHS directive 2011 65 EU Restriction of Hazardous Substances FCC 47 CFR 15 unintentional radiator class B Digital Devices Attention Use manufacturer supplied cable s only to connect all I O s with other devices Thoroughly connect the shielding to common Manufacturer will not accept any liability for damage direct or indirect caused by connecting this instrument to devices which do not meet relevant safety standards WARNING March 26 2015 RoHS User manual AS110 edition 7 Intended use For research purposes only While examples of clinical applications may be shown this instrument is not tested by the manufacturer to comply with the In Vitro Diagnostics Directive WEEE directive Antec is a Business to Business producer of analytical analysis equipment which fall under WEEE Annex IA categories 8 and 9 includes medical devices and monitoring and control instruments All equipment of Antec Leyden which are subjected to the WEEE directive shipp
59. n 7 analysis In the example it is set to 10 minutes All other values are the default values Method Setup example ha IV Enabled Figure 22 Flow path and aspiration settings in System settings tab Method Setup example 0900000 900000 000000 9090000 000000 900000 89090000 0090000 Figure 23 Tray configuration settings in Tray tab Click Ok and re save the method file to store all changed settings Analysis of the sample can be achieved by programming a sample sequence with a single line Note that the option single analysis in Clarity is intended for injections with a manual sampling valve Chapter 4 53 e Make sure that sample tray position A1 left contains a sample vial e Open anew sequence by clicking the sequence icon in the method window e Program one line with SV 1 start vial and fill in all other relevant parameters sample ID sample inj Vol 5 uL method name etc e Click start to execute the sequence RE File Edit Sequence View Window Help L fl TAL w El ES BETTE Inj Vol Method Report Partial loopfill 5 000 0 q No example Instrument Of a es For Help press F1 Single Run Not ready no method sen Vial f Inj File Name Active A Figure 24 sample sequence for a single injection of 5 uL sample with partial loopfil method Example 2 three full loop injections 10 uL including wash
60. n of more than 2mm towards home Home sensor not de activated Home sensor activated when not expected Tray position is unknown Horizontal needle position is unknown Horizontal home sensor not reached Horizontal home sensor not de activated Horizontal home sensor activated when not expected Vertical needle position is unknown Vertical home sensor not reached Vertical home sensor not de activated Vertical stripper did not detect plate or wash waste Vertical stripper stuck Vertical The sample needle arm is at an invalid position Syringe dispenser unit Syringe valve did not find destination position Syringe home sensor not reached syringe home sensor not de activated Syringe position is unknown Syringe rotation error Destination position not reached Wear out limit reached Illegal sensor readout Chapter 8 Cooling unit 77 ERROR 347 Temperature above 48 C at cooling ON Electronic ERROR 280 ERROR 282 ERROR 283 ERROR 284 ERROR 290 EEPROM write error EEPROM error in settings EEPROM error in adjustments EEPROM error in log counter Error occurred during initialization the AS 110 cannot Start 78 User manual AS110 edition 7 Chapter 9 CHAPTER 9 Specifications General Sound pressure level Working temperature Storage temperature Humidity Safety and EMC compatibility Installation class Pollution degree Altitude Dimensions Fr
61. n the Initial wash group box to perform a standard wash routine All tubing connected to the syringe valve will be rinsed with wash solvent Click Close to leave the Direct control screen The AS 110 is initialized In case an air bubble is trapped at the tip of the syringe or in the tubing perform these steps with the wash solvent tubing end in 100 methanol When the bubble is removed repeat again with the final original wash solvent Sample handling Take the following into account when handling samples Standard vials can best be filled by means of a narrow end pipette to allow air to escape when filling the vial Do not fill vials wells to the edge If you do sample will be forced into the air needle risking cross contamination of samples and soiling the needles It is important that seals and capmats are airtight to prevent air bubbles from forming and to block evaporation of volatile samples We recommend use of the following seal types o for standard low well plates sealing tape o for deep well plates pierce able capmats Pre slit or silicon or sealing tape o for vials standard septa thin types do not use vials with hard caps that are not designed for being pierced by an 42 User manual AS110 edition 7 injection needle do not use e g Eppendorf SafeLock micro test tubes When you use uncapped vials wells injection performance may not be to specification Filtering the eluent with 0 2 um filter will considerably
62. n the rear side of the instrument match that for the power outlet to be used Understanding power surges Power surges line spikes and transient energy sources can impede instrument operation Ensure that the instrument s electrical supply is adequately protected from these conditions and properly grounded Chapter 2 35 6 Connect the AS 110 to the PC COM port with the cable provided with the AS 110 7 Connect the power cable between the AS 110 and the power outlet Power supply and protective earth The system must be connected to a suitable mains power supply with a correctly installed protective earth conductor Never use the system without a properly connected protective earth conductor ELECTRICAL SHOCK 8 Switch on the AS 110 9 On your PC open AS 110 Service Manager and enter the required settings 10 Connect the drain tubing to the waste outlet 11 Fill the wash solvent bottle inside the sampling compartment of the AS 110 with distilled water and propanol 80 20 v v or mobile phase Only water or organic solvents should be used Do not use crystalline or buffer solutions as these may block the system and cause severe damage Degas the wash solvent to prevent air bubbles from forming in the syringe 12 Fill the wash solvent tubing syringe and buffer tubing by washing the system two or three times Use 100 IPA for better degassing or removing of air bubbles 13 Check if air bubbles are trapped in t
63. needle by clicking Start in the Initial wash group box of Direct Control 10 Use the Autosampler Adjustments option to adapt Needle Tray settings If you use trays with 12 vials or 48 vials make sure that the needle height settings is gt 2mm to prevent the needle from touching the bottom of the vials CAUTION Chapter 5 59 Air needles for AS 110 Six types of air needles are available for the AS 110 autosampler all different in length difference of 6 mm These air needles are required to accommodate use of different plate heights in the AS 110 For every well vial plate the correct air needle is available Apart from the 6 mm difference in length between the air needle types the needle holder allows for an extra 6 mm variation in needle height Standard air Needle k A 2 mL vial Greiner deep well with capmat Greiner low well The standard air needle is a 62 mm needle no 0045 505 This air needle accommodates use of a wide range of high and low plates See the illustrations above for the puncturing depth of the needle 60 User manual AS110 edition 7 Note that no PASA should be used for low wells as the sample needle sufficiently punctures the seal to prevent vacuum the function of the air needle will be insignificant for the low well plates If the 10 mL vials are used the air needle is lowered pretty far into the vial If the vial is not f
64. not accept any liability for damages directly or indirectly caused by connecting this machine to instruments which do not meet relevant safety standard The IO connector contains active high or active low TTL inputs and one contact closure output user definable in the System Settings The two inputs can be programmed as Next Injection Input Freeze Input or Stop Input The Next Injection Input Freeze Input and Stop Input can be used to control the AS 110 by other devices The contact closure output can be programmed as nject Marker Auxiliary or Alarm output Table 3 IO connector Contact closure output and TTL inputs Pin no Description Cable colors Output Common RED 3 wired Output Normally open BLACK 3 wired Input 1 RED 4 wired Input 2 BLACK 4 wired GND Output Normally BROWN 3 wired closed GND GND ORANGE 4 wired GND BROWN 4 wired Contact closure output e Inject Marker Output default an Inject marker output will be generated when the injection valve switches from LOAD to INJECT Status duration of the Inject Marker is the same as for the setting for the SparkLink Inject marker pulse Range of the adjustment of the inject marker pulse is 0 1 2 0 seconds Chapter 7 75 e Alarm Output the Alarm Output will be activated whenever an error occurs see appendix C for a description of the error codes of the AS 110 e Auxiliary the contact closure output can be used as an Auxiliary which can
65. ns and treat all soecimens potentially infectious Always wear protective and gloves when handling toxic or biologically infectious samples to prevent bio hazards or hazards while working with the Alias Waste disposal Do not allow flammable and or toxic solvents to accumulate Follow a regulated approved waste disposal program to dispose waste sample solvents device Never dispose of flammable and toxic solvents or sample through the municipal sewage system Applications quality control It is recommended that you routinely run several quality control samples Quality control samples should represent low average and high levels of a compound Make sure that quality control sample results are within an acceptable range and evaluate precision from day to day and run to run Data collected when quality control samples are out of range may not be valid Do not report this data until you are certain that system performance is acceptable Apart from use of quality control samples we recommend that you use blanks The blanks will help you assess whether carry over is within an acceptable range and monitor the integrity of your data Using the AS 110 in other ways than indicated in the manual or defined by good laboratory practice may result in unsafe operation vi User manual AS110 edition 7 Table of contents DECLARATION OF CONFORMITY i Intended use ii WEEE directive ii ROHS directive ii About this manual iii Symbo
66. nsitive online help is available with every window and dialog Special attention has been paid to ensure a service friendly design To enhance safety speed of operation of the AS 110 will decrease when the door is opened Optional sample cooling ensures consistent results Read this chapter to help identify parts of the AS 110 auto sampler and to learn more about injection principles User manual AS110 edition 7 Autosampler configurations There are differently configured AS 110 autosampler with the details listed in Table 1 All replaceable parts are easily accessible Refer to the List of accessories and spares Appendix I for more information Table 1 Different AS 110 autosampler configurations with details of installed parts Abbreviations St standard m micro 6p 10p 6 10 port U UHPLC T pae O L 3 a g D 3 3 la S 0 ab Cc O Q Q t D 5 5 O O D O Cc a 5 a a S a 2 z 2 3 E 2 P E f gt gt 5 Q O U gt ep U Mm 0 191 0035 St 6p 1 16 15 500 1000 100 191 0035u St 6p U 1 32 15 250 200 5 191 0036 St s 6p 1 16 15 500 1000 100 191 0037 m 6p 1 16 2 4 25 200 20 191 0037u m 6p U 1 32 2 4 25 50 5 191 0038 m 10p 1 16 2 4 25 200 2 2x 191 0038u m 10p U 1 32 2 4 25 50 2 2X 191 0041 St 10p 1 16 15 500 1000 5 2x 191 0041u St 10p U 1 32 15 250 200 5 2x Chapter 1 Instrument description AS 110 autosampler front The AS 110 sampling com
67. on Tray position Tray Home Set Springe position Home Set Valve position Inject Set Chapter 3 45 Configuration in Clarity The appropriate AS 110 control module under the AS modules can be selected and configured in the Clarity configuration menu FEE ERE Fite A A Name Vendor Module Info Comment AS El 1100 ALS Agilent G13134 G13294 G1330B as part of 1100 1200 syste ESA Grace Alltech Grace Alltech CTC Spark Holland Developed by Knauer Dani Shimadzu A40C 20i AOC 20s AOC 20d 4S 100 Antec 45110 Antec Developed by Knauer AS 3800 Knauer 4S 3900 Knauer 4S 3950 Knauer Developed by Knauer AS 54 Ecom 452000 Analptical Tech Endurance Spark Holland HP 7673 Agilent HP 7673 HP 7673 II HP 7683 in emulation mode HP 7683 Agilent part of 6890 HT 200H HTA HT 200H HT 250D HT 2807 HT 3004 HTA Euro 45300 HT 300L HT 3104 HT 310L HT 2807 HT 800L HTA Developed by HTA LaChrom Elite 4S Merck Hitachi L 2200 as part of LaChrom Elite system LC 10 20 45 Shimadzu SIL 104Dvp SIL HTA SIL HTC SIL 204 SIL 204C Marathon Spark Holland al Lana Aidaa Camel bere al Ra Figure 14 Available AS control modules in the Clarity configuration The AS 110 micro control module is highlighted in blue System Configuration i x Setup Control Modules Number of Instruments AA Instrument 1 xe Instrument 2 x Instrument 3 x Instrument 4 F as O Antec 45 110 micro Instrument Ty
68. partment houses the following parts Figure 1 AS 110 sampling compartment Syringe Needle arm Injection valve Valve leak bin Sample compartment Needle wash position Wash liquid bottle SM S DE he IN User manual AS110 edition 7 Tubing guide Wash waste Condensed water leakage Cooling cover ey s Figure 2 AS 110 with cooling front side AS 110 autosampler back The back of the autosampler has the following items 1 2 3 Figure 3 AS 110 with cooling back side 9 pin male connector inputs output 9 pin female connector serial interface warning label see I O connections on page 74 on off switch fuse box power connector cooling fan if cooling option is installed do not obstruct type label oo SE Sh SN r Chapter 1 Access To open the door 1 Get hold of the door handle 2 Gently pull it towards you and push it upward until it is in horizontal position 3 Slide the door into the autosampler Figure 4 Opening the door For easier access to e g the syringe the whole front cover of the AS 110 can be removed figure 4 To remove the cover 1 Press the two black buttons on either side top of the autosampler simultaneously 2 Gently pull the cover towards you N Figure 5 Removing the AS 110 front cover The black arrows indicate the location of black push buttons Right picture shows the AS
69. pe SN Sampler 1 Instrument 1 fic 7 Name Example LC EI Gc Image For Closed Instrument Image for Opened Instrument H O Detector MS Balance Thermostat Fy alve Fraction Collector Capillary i Electrophoresis i Auxiliary Antec 45 110 micro iL Cane Detector MS Z Thermostat alve Fa Fraction Collector Auxiliary Device Number Ext Start Dig Input Jas 110 micro autosampler ie In Fa Ready Dig Output Add Remove About Setup Cancel Help Figure 15 System configuration with AS 110 control module configured and added to LC instrument 1 in Clarity 46 User manual AS110 edition 7 Serial Humber fi 0300 ok Communication ID EH Cancel Interface AS 232 si COM Part Jcomio Figure 16 In the AS 110 control module set up screen the corresponding COM port can be selected To add an AS 110 auto sampler to a LC instrument in Clarity and configure it please follow the steps below e Add the AS 110 control module to the Set up control modules box in the Clarity configuration window using the add button e Double click on the AS 110 control module to access the set up menu and set the appropriate COM port e Click the to check the communication with the connected AS 110 In case communication is established a new box will open with serial number device and Firmware version information Close the box using the Close
70. rammed at the desired position to activate the compressor to put air pressure on a sample The compressor will stay active until it is switched off in a next programmed step The compressor will be automatically switched off at the end of the needle wash routine if a needle wash is used to control the standard auxiliary see Control I O connections contact closures to program a pause in which the Sineas waits for one of the two inputs see Control I O connections to become lt HIGH gt or lt LOW gt before continuing with the next step to control the marker only possible when output is programmed as Inject Marker see Control I O connections option to define the Solvent Selection Valve SSV port position range 1 to 6 Note the total number of steps for the user program and the mix method cannot exceed 240 Chapter 1 r 2 3 4 5 6 7 8 9 LOW NORMAL HIGH SYRINGE 25 UL 3 uL min 13 uL min 31 uL min 62 uL min 94 uL min 192 uL min 267 uL min 343 uL min 436 uL min 50 UL 6 uL min 25 uL min 63 uL min 125 L min 188 uL min 384 uL min 533yuL min 686 uL min 873 uL min 100 uL 13 uL min 50 uL min 126 uL min 250 uL min 377 uL min 768 uL min 1067 uL min 1371 uL min 1745 uL min During the dispense action the pressure in the buffer tubing will increase To prevent damage of the syringe valve the flow should not exceed the value of 250 L min for water Maximum speed 6
71. rapping of the equipment BIOHAZARD General instructions for disposal When taking the instrument out of service the different materials must be separated and recycled according to national and local environmental regulations Hazardous substances The instrument contains hazardous substances Contact the manufacturer for more detailed information Disposal of electrical components Waste of electrical and electronic equipment must not be disposed as unsorted municipal waste and must be collected separately Please contact an authorized representative of the manufacturer for information concerning the decommissioning of equipment INDEX u uL pick up 53 uL Pickup injections 18 A Accessories amp Spares 83 Add 25 Air in flow path 71 Air needle replacement 62 Air needles 8 Air needles for Alias 8 Air needles for AS 110 59 Air segment with uL Pickup injections 21 Air segment with Full loop injections 14 Air segment with Partial loopfill injections 17 Alarm output 74 ALEXYS control menu s 43 analytical problems 69 Analytical problems 94 AS 110 fluid connections 37 AS 110 tubing 38 ASM software 35 B Black push buttons 5 C Calibration amp Performance 87 carry over 88 Cleaning 55 Contact closure output 74 Contact closures output and TTL inputs 74 Cooling 8 Cooling unit 77 D danger sign iii Dead volumes in tubing connections 71 DECLARATION O
72. roducibility or carry over may occur in any HPLC system It may be hard to find the cause you may have to try out several procedures The first thing to do is to determine whether the problem is caused by the autosampler or by the rest of the system Chapter 6 71 1 Replace the valve by a manual injection valve to discriminate between valve problems and other problems 2 Doanumber of Full loop injections If the results are fine the fault is in the autosampler if not check the rest of the HPLC system Please bear in mind that analytical problems may also be caused by external influences like temperature or light sensitive samples Make sure that the application was running trouble free before and that no changes have been made to the system A number of causes and possible solutions for analytical problems is listed below Contact service if you need further help If reproducibility is not according to specifications check the following possible causes Causes Solutions Air in flow path Do an initial wash select AS 110 Direct Control in AS 110 Service Manager Leaking syringe If leakage occurs at the top of the syringe check whether it has been properly mounted If leakage occurs at the bottom of the syringe replace plunger tip or syringe Leaking syringe Check or replace valve valve Rotor seal worn out Replace seal Check stator Syringe valve seal Check if installed Dead volumes in Redo connections with new f
73. s also removed through this tubing e Condensation water and leakage drain through the left hand hose connector see Figure 2 number 3 all leaked solvents and condensation from cooling are drained If the cooling is used you are advised to connect this hose connector in the ship kit of the AS 110 to a waste container on the floor e Make sure that none of the drain or waste tubes is twisted this might obstruct the flow path Wash solvent and syringe rinse Use aclean bottle for the wash solvent and place it on the left hand side of the AS 110 You are recommended to use electrochemically clean water Chapter 2 41 R lt 18MOhm cm or a mixture of such water and iso propanol 80 20 as wash solvent Before using the wash solvent degas the solvent with helium or an ultrasonic bath Do not use salts or buffer in this solution crystals may block or damage the system To fill the wash solvent tubing execute the following steps i Place the end of the wash solvent tubing in the filled wash solvent bottle Open Direct Control in AS 110 Service Manager In the Syringe group box click End A syringe volume of wash solvent is aspirated from the wash solvent bottle and the wash solvent tubing is filled Click Home The syringe contents is dispensed to syringe waste Repeat steps 3 and 4 until the wash solvent tubing and the syringe are completely filled When wash solvent tubing and syringe are completely filled click Start i
74. steel Valco C2 2006 181 0332 Rotor seal for Valco C2 2006 181 0336 Stator for Valco C2 2006 181 0362 Valve stainless steel Valco C2 1006 181 0368 Rotor seal for Valco C2 1006 181 0372 Stator for Valco C2 1006 181 0392 Valve stainless steel Valco C2 1000 181 0388 Rotor seal for Valco C2 1000 181 0390 Stator for Valco C2 1000 191 0352U UHPLC valve C72NX 6676 191 0354U Rotor seal UHPLC valve C72N 191 0356U Stator UHPLC valve C72NX 6676 UHPLC micro UHPLC micro UHPLC micro UHPLC std FW gt 1 26 std micro UHPLC micro UHPLC std FW gt 1 26 std all 6 p std 6 p std 6 p std 6 p micro 6 p micro 6 p micro 10 p micro 10 p micro 10 p micro 6 p UHPLC 6 p UHPLC 6 p UHPLC APPENDIX A 85 description 250 1200 Loop 5 ul SS Valco C type 1 16 std micro 250 1201 Loop 2 uL SS Valco C type 1 16 std micro 250 1202 Loop 10ul SS Valco C type 1 16 std micro 250 1204 Loop 20ul SS Valco C type 1 16 std micro 250 1206 Loop 50ul SS Valco C type 1 16 std micro 250 1208 Loop 100ul SS Valco C type 1 16 std micro 250 1214 Loop 1 mL SS Valco 1 16 std micro 250 1220 Loop 1 5 uL SS Valco 1 32 UHPLC 250 1222 Loop 2 uL SS Valco 1 32 UHPLC 250 1224 Loop 5 uL SS Valco 1 32 UHPLC 250 1226 Loop 10 uL SS Valco 1 32 UHPLC Tubing amp connections AS flow path description 191 0320 Buffertubing 1000uL Tefzel std 191 0320M Buffertubing 200uL Tefzel micro 191 0334 AS 110 tubing set std 191 0334M AS 11
75. t prevents correct uploading of the new files This problem may be solved by adapting the size of the serial ports FIFO Chapter 2 37 Execute the following steps to adapt settings w co AL O 01 Open the Windows Device Manager Configuration settings System settings Communication port In the COM port properties select the Port settings tab Click Advanced Use the sliders to change the size of the receive buffer The default values for Receive buffer high 14 and Transmit buffer high 16 should be sufficient However decrease the values to 8 if buffer overflow errors appear Click OK and close the Windows Device Manager Restart your PC Open ASM and try to upload again Contact service if there still are problems during uploading AS 110 flow path Refer to Table 1 for the details of the factory installed items in the flow path for an AS 110 autosampler with a 6 port valve Figure 11 and a 10 port valve Figure 12 Notice that the waste line is not connected to the left side port of the syringe valve but on the backside port which is not visible and accessible without loosening removing the syringe valve Note with the AS 110 standard FW gt 1 26 it is possible to use the left side port of the syringe valve to connect a second wash bottle supported in the AS 110 Clarity driver developed by DataApex Spark Pump Column C 7 INJECT position Wy LOAD position Bottle Figure 11 AS 110 flui
76. tap the syringe as the wash solvent is dispensed to syringe waste Perform a standard wash routine Direct control click Start Stop in de Initial wash group box All tubing connected to the syringe valve will be refilled and flushed Make sure the wash is compatible with IPA otherwise use a different solvent before switching to the wash solvent syringe plunger amp plunger tip Execute the following steps to replace the plunger or plunger tip In Direct control click Exchange in the Syringe group box Remove the syringe Slide the plunger out of the glass part of the syringe With pliers remove the tip Dampen the new tip with for example isopropanol Mount the new tip on the plunger Insert the plunger in the glass part the syringe Install the syringe in the autosampler again at Oo ee TO Replacing the Syringe dispenser valve The syringe valve is a 4 port selection valve Ports are assigned as follows Waste Use this port as a drain for the syringe dispenser Wash Use this port to aspirate wash liquid from the wash bottle or in case of multiple wash liquids connect it to the solvent selection valve Needle Connect the buffer tubing to this port All connections to the syringe valve must be made using fingertight fittings An exception can be made for the waste outlet the port on the rear of the valve Execute the following steps to replace the syringe dispenser valve Chapter 5 65 Place t
77. the sample position e Wait the programmed period of time before continuing with the next step reaction time H MM SS maximum of 9 hours 59 minutes and 59 seconds A maximum of 15 steps can be created for the Mix amp Dilute routine Sample positions Mix amp Dilute Q If a mix is programmed the reagent solvents destination and sample positions in the trays are as follows e Reagent A Left plate e Reagent B Left plate e Samples Right plate e Destination Right plate If you chose to process plates in rows the following positions are available for sample destination reagent A and reagent B OOOO 0000 OOOO 0 000 OOOO 0 00 OOOO 0 000 OOOO 00 0O OOOO 0 00 OOOO 00 0O OOOO 0 000 OOOO 0 000 OOOO 0 000 OOOO 0 000 OOOO 0 000 OOOO AIAI OOOO AIAI OOOO AIAI OOOO LAI AA OIOIOIOL SL AL AL S OOOO AL AL AL A OOOO AL AL AL A OOOO AIAI OOOO LAII OOOO LAII OOOO AIAI COOOL IAI For example you can program the following e two 96 low plates processed in rows e first line first sample position 2A1 last sample position 2A1 destination position 2E1 Reagent A 1A1 and Reagent B 1E1 e second line first sample position 2A2 last sample position 2A2 destination position 2E2 Reagent A 1A2 and Reagent B 1E2 Sample 7 Destination O Reagent A Reagent B Chapter 1 27 e third line first sample position 2A3 last sample position 2A3 destination position 2E3 Reagent A 1A3 Reagent
78. tion volume 0 uL 19 999 uL with 1 uL increments Valve Valco 0 75 bore valve Sample loop 10 mL SS sample loop 1 8 tubing with 1 16 tubing ends and fittings Valco Buffer tubing 2mL Needle LSV needle with LSV air needle Promis and seal Analytical performance Injection method Partial loopfill injection mode Reproducibility RSD lt 1 0 for partial loopfill injections injection volumes gt 10 UL up to 50 of the installed sample loop Viscosity range 0 1 5 cP Memory effect lt 0 1 with programmable needle wash APPENDIX A 83 APPENDIX A Accessories amp Spares The ship kit of the AS 110 contains the following parts 191 0306 191 0552 191 0512 191 0522 181 0590 181 0592 191 0528 181 0578 181 0356 191 0322 191 0340 191 0342 191 0328 191 0300 Description Air needle 80 mm low format well plates CD Rom ASM en users guide I O Cable Tubing connector T piece power cord EUR power cord USA Cable Multilink Fuse 2 5 AT Valco shipkit PP wash solvent bottle 250 mL rectangular AS 110 wash bottle adapter AS 110 wash bottle 250 mL glass Silicone tubing 1 meter 48 position vial adapter NO AS 110 micro is equipped with a glass wash bottle and wash bottle adapter The following parts are available for the AS 110 191 0200 191 0200M 191 0322 191 0328 191 0340 191 0342 191 0512 191 0522 191 0528 191 0530 191 0532 191 0550 191 0556 191 0600 191 0602 191
79. tosampler for execution of analytical runs An index allows the user to find required information quickly Symbols The following symbols are used in this guide The danger sign warns about a hazard It calls attention to a procedure or practice which if not adhered to could result in injury or loss of life ELECTRICAL SHOCK Do not proceed beyond a danger sign until the indicated conditions are fully understood and met The warning sign denotes a hazard It calls attention to a procedure or practice which if not adhered to could result in severe injury or damage or destruction of parts or all of the equipment Do not proceed beyond a warning sign until the indicated conditions are fully understood and met The caution sign denotes a hazard It calls attention to a procedure or practice which if not adhered to could result in damage or destruction of parts or all of the equipment Do not proceed beyond a cautions sign until the indicated conditions are fully understood and met CAUTION The biohazard sign draws attention to the fact that use of biological materials viral samples and needles may carry a significant health risk BIOHAZARD The attention sign signals relevant information Read this information as it might be helpful The note sign signals additional information It provides advice or a suggestion that may support you in using the equipment User manual AS110 edition 7 Safe
80. ts three tubes NEEDLE connection to sample needle WASH connection to wash solvent bottle WASTE connection to syringe waste tubing to control the action of the syringe LOAD the syringe with the programmed volume UNLOAD the syringe with the programmed volume HOME the volume previously aspirated will be dispensed to the last programmed position and the syringe will be initialized again to execute a needle wash the content of the buffer tubing is not rinsed to waste before the start of the wash The programmed volume of wash solvent is used to wash the needle at the wash position The wash position may be contaminated with the contents of the buffer tubing which may generate cross contamination To prevent contamination of the wash 30 VALVE WAIT COMPRESSOR AUXILIARY WAIT FOR INPUT MARKER SSV User manual AS110 edition 7 position program a dispense to waste action before programming a wash action to program positions of high pressure valves ISS or injector valve The injector valve has two positions lt INJECT gt and lt LOAD gt The ISS optional valve has positions 6 1 and 1 2 Figure ISS valve positions to program a pause max 9 hours 59 minutes 59 seconds During the pause the needle will move to home position if the previous step is an aspirate or dispense action If you want the needle to stay in the same position an aspirate or dispense step of 0 uL must be prog
81. ty instructions Adhere to the following standard quality control procedures and the following equipment guidelines when using the auto sampler The following safety practices will ensure safe operation of the auto sampler and should only be executed by authorized service personnel Working environment amp safety The intended use of the system is to introduce sample into a mass spectrometer or other U HPLC detection system in a GLP approved environment Operators using the system should have the appropriate warnne education an extensive understanding of GLP rules and be skilled in the art Use this system ONLY for the intended use Use of the system for any other purpose will cause unsafe situations System Operation To keep up the high performance of the autosampler we recommend that the system is checked regularly and maintenance procedures are carried out Preventive maintenance contracts are available for that purpose Please contact your local dealer or the nearest sales office for more information Electrical safety Removal of panels may expose users to dangerous voltages For that reason this should only be done by authorized service engineers Disconnect the autosampler from all power sources before removing ELECTRICAL SHOCK protective panels Replace or repair faulty insulation on power cords immediately after discovery of the fault Check that the actual power voltage is the same as the voltage for which the instruments
82. ubing 40 WEEE ii working order 45
83. with a 25 uL syringe but a 50 uL or 100 uL syringe can also be installed Chapter 5 63 Figure 26 Syringe Execute the following steps to install a different syringe 1 In Direct Control click Exchange in the Syringe group box 2 Unscrew the syringe from syringe valve but make sure that the connector in the valve remains in place 3 Disconnect the plunger from the syringe drive 4 Fill the new syringe with wash solvent preferably IPA iso propanol Make sure that most air bubbles are removed from the syringe In case of a 25 uLsyringe priming with 100 MeOH may be necessary for air bubble free installation 5 Connect the plunger of the filled syringe to the syringe drive and connect the syringe with the connector at the syringe valve Insert the end of the plunger well in the plunger lock if it is not well inserted the glass syringe will break when the plunger is moving to the home position CAUTION 6 Screw the syringe firmly into the bottom port of the syringe valve 7 In Direct control click Home in Syringe group box The syringe moves to home position and its content will be dispensed to syringe waste 8 If there is still some air in the syringe click End again in Direct control The syringe is filled with wash solvent Use IPA 9 Click Home again to dispense the wash solvent to waste 64 User manual AS110 edition 7 lf there is still air in the syringe repeat steps 8 and 9 and gently

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