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Chap0 - FAMOS Well Plate Microautosampler

1. y FIGURE 6 4 Troubleshooting the Microautosampler Note It is important to note that analytical problems might be caused by external influences such as the temperature and or the analysis of light sensitive samples When troubleshooting make certain that the application was running in an acceptable fashion before the problems were observed and that nothing has been changed in the application D931R1 User s Manual FAMOS Maintenance and Troubleshooting 6 5 Error Codes of the Microautosampler The FAMOS Well Plate Microautosampler will display an error message if the user tries to enter invalid data and information on the allowed range will be displayed In addition several sensors are installed to check for possible mechanical failures and protect the instrument from severe damage If a fault or mechanical failure is observed during the operation of the FAMOS Well Plate Microautosampler an error code will be presented on the display When a message is indicated press the Start Stop key twice to remove the message and correct the fault The error codes are indicated in TABLE 6 6 to TABLE 6 11 TABLE 6 6 Error Codes for Injection Valve and ISS Unit Probable Cause Solution ERROR 11 Injection valve is not in a valid position Contact A ERROR 12 The injection valve did not switch within 1 5 Contact Ej seconds LC Packings ERROR 13 The switching time of the injection valve Contact ERROR 14 ISS
2. Caution Make certain that the fuse holder is inserted in the correct position so that the appropriate voltage setting is indicated FIGURE 6 3 indicates the configuration for 220 240 V User s Manual FAMOS D931R1 6 9 Maintenance and Troubleshooting 6 4 Troubleshooting Troubleshooting refers to the determination of the cause of a problem Since the FAMOS Well Plate Microautosampler is incorporated into an HPLC system the first step to determine if the problem is due to the microautosampler is to remove the unit from the system install a manual injector and then perform an injection and compare the results from the two runs If the results are fine the problem is due to the autosampler Analytical problems also might be caused by external influences like temperature and or light sensitive samples For this reason it is important to be sure the application was running without problems before and nothing has been changed A chart outlining the most common faults and the remedies is presented in FIGURE 6 4 BAD REPRODUCIBILITY Air in Flow gt Path Start a manual wash Replace plunger tip or syringe Leaking Syringe y Leaking Syringe Valve Replace Valve Rotor seal worn out Replace rotor seal Check Stator face y Dead volumes in tubing connections Redo connections with new ferrules and nuts
3. Autosampler Output A o GND Ext device Input TTL GND FIGURE E 2 TTL outputs E 3 1 P2 Connector TTL OUTPUTS TABLE E 4 P2 Connector Pin Descriptions PNa Function Pns Function INJECT MARKER o Inotconnected 6 not connected 14 Signal ground Signal ground 8 _ motconnected do To oo All markers are active low logical O Vmax 5 5 V logical 1 gt 3 5 V logical O lt 1 0 V DC output source sink current 20 mA IE Note A marker output pulse will be generated when the injection valve switches from LOAD to INJECT However in a User Program markers have to be programmed by the user E 3 2 P3 Connector TIMED OUTPUTS TABLE E 5 P3 Connector Pin Descriptions Connector P3 TIMED OUTPUTS 4 bit time base code output PIN 1 TBO HEX 1 6 Signal ground _ _ TB 1 HEX 2 Signal ground TB 2 HEX 4 8 Signal ground 4 TB3 HEX 8 9 Signal ground _ 5 motused S O S All markers are active low logical O Vmax 5 5 V logical 1 gt 3 5 V logical O lt 1 0 V DC output source sink current 20 mA E 4 D931R1 User s Manual FAMOS Digital Inputs and Outputs E 4 TTL Inputs The TTL input connector FIGURE E 3 is an active high or active low TTL input it can be defined in the System Menu The NEXT INJECTION INPUT and the NEXT WELL INPUT can be used when the FAMOS Well Plate works in REMOTE CONTROL The FREEZE INPUT
4. In addition a 96 deep well plate is used and some wait steps are included to increase the performance a Place a sample vial in position A 01 place an empty sample vial in position B 01 as destination vial Place a filled reagent vial in position 1 Make sure the reagent vial is filled correctly before starting a new series b Program the System Parameters by entering the parameters indicated in TABLE C 9 TABLE C 9 Programming System Parameters System to enter the System Menu lt CONVENTIONAL gt E to select the conventional mode lt USAGE gt E enter the Usage Menu EE go to the Mix field lt ENABLED gt to enable use of mix methods Escape return to the System Menu lt PLATES gt E to enter the Plates Menu lt 96 HIGH gt to select 96 deep well plate lt COLUMNS gt to select processing in columns A1 B1 etc CL to disable use of transport vials Escape Escape to return to the Ready Menu Note As soon as a change has been entered in the System settings the message ALL SERIES DEFAULT appears The user will have to redefine series because the settings have been changed c Program the method by entering the parameters indicated in TABLE C 10 TABLE C 10 Programming Methods Parameters Methods to enter the Methods Menu lt INJECTION gt 03 E to enter the Injection Menu lt PARTIAL gt E to select partial loopfill injection mode 100 E to define the analysis time 50 E to define the flush volume
5. a Syne C KA Coum o gt FIGURE A 2 Sample Line Filled with Sample When the injection valve is switched into the LOAD position FIGURE A 3 a sharp sample front is placed at the inlet of the sample loop lt Syringe f Columa gt mA ANANN H iN E Sample AY FIGURE A 3 Injection Valve Switched into LOAD Position For full loop injections the sample loop is quantitatively filled by transporting two or more times the loop volume through the loop FIGURE A 4 The actual volume that is transported is dependent on the volume of the loop FIGURE A 4 Transporting Two or more Times the Loop Volume Through the Loop The injection valve is then switched into the INJECT position FIGURE A 5 The sample loop is now part of the HPLC mobile phase flow path so that the sample is transported to the column and the analysis time starts A 2 D931R1 User s Manual FAMOS Injection Principles Ds A EZ Syringe fi BA gt Sample FIGURE A 5 The Injection Valve Switched into the INJECT Position If one injection is to be done from each well or if a wash routine has to be performed after every injection the needle withdraws from the well directly after the injection and immediately washed if programmed After the analysis time is completed a new sequence is started If more than one injection is done from the same well without a wash routine the FAMOS Well Plate Microautosampler withdraws a flush vol
6. If the mobile phase includes volatile or flammable solvents make certain that they are not allowed to enter the workspace 2 If the mobile phase includes volatile or flammable solvents avoid open flames and sparks 3 If aleak occurs turn off power to the instrument and remedy the situation immediately 4 All components of the system should be plugged into a common power line that is directly connected to a true ground 5 When the panels are removed dangerous electrical connections will be exposed Disconnect the autosampler from all power sources before removing the panels 6 Always replace blown fuses with fuses of the same size and rating indicated on the fuse holder and panel Refer to Section 6 3 6 of this manual for more information on Fuses User s Manual FAMOS D931R1 iX Warnings and Safety Precautions 7 Repair or replace faulty power cords and all communication cables 8 Many organic solvents and buffers are toxic Make certain that you know the toxicological properties of all mobile phases that you are using 9 The toxicological properties of many samples may not be well known If you have any doubt about a sample treat it as if it contained a potentially harmful substance 10 Wear protective eye goggles when handling mobile phases or operating the instrument An eye wash facility and a sink should be close to the unit If any mobile phase splash on the eyes or skin wash the affected area and seek medic
7. Note When lifting the FAMOS Well Plate Microautosampler from the shipping container make sure that the unit is kept upright Lift the unit by placing your hands under the microautosampler If there is external damage to the shipping box the damage should be reported to the shipping agent and LC Packings upon receipt of the goods If internal damage is observed or if any items are missing this should be reported to the shipping agent and to LC Packings as soon as it is observed Note If there is any apparent damage to the instrument the user should investigate the nature of the damage before plugging the unit into the mains to ensure that powering up of the instrument will not create a hazardous condition or damage internal components If the damage appears significant call LC Packings or its local representative before connecting the unit to the mains 2 2 D931R1 User s Manual FAMOS Installation and Getting Started 2 3 The Standard Microautosampler Configuration A number of components of the FAMOS Well Plate Microautosampler are factory installed as presented in TABLE 2 1 The standard tubing configuration is presented in TABLE 2 2 If desired these components can be replaced to meet the specific needs of the analysis a complete list of accessories and consumables is presented in Section 6 6 Changing the components is described in Section 6 3 Refer to FIGURE 2 1 to identify the various components TABLE 2 1 Facto
8. b Place a thermocouple in the middle of the cooling plate flat surface underneath the wash solvent bottle item 3 FIGURE 2 5 making sure that a good contact is made c Switch on the cooling and program a setpoint of 10 C d Wait minimal 15 minutes for equilibration of the FAMOS Well Plate Microautosampler e Read out the temperature of the thermocouple The value must be in a range of 2 C of the programmed setpoint IBS Note When the cooling option is used the temperature of the sample inside the sample vial may be slightly different than the temperature set via the command due to a variety of effects e g the heat transfer characteristics of the vial walls If it is necessary to precisely set the temperature of the sample we recommend that you determine the temperature inside the vial at various temperature setpoints and set the cooling option so that the desired internal temperature is attained 5 6 D931R1 User s Manual FAMOS CHAPTER 6 Maintenance and Troubleshooting Overview This chapter provides information to assist in optimizing the performance of the FAMOS Well Plate Microautosampler and maintaining it in your laboratory It includes the following material Maintenance describes a series of activities that should be performed on a periodic basis to optimize the performance of the system and minimize down time Section 6 2 Replacing Components provides directions for replacing com
9. until all series is completed e NEXT WELL INPUT this input will start the first injection from the next vial in the series If the series was completed it starts the first injection from the first vial of the next series until all series are executed eS Note The difference between the NEXT INJ ECTION and the NEXT WELL INPUT is that the NEXT WELL INPUT directly starts the first injection from the next vial even if not all programmed injections from the previous vial were executed To indicate that the remote control mode is activated the letter r is displayed in the lower left corner of the display To execute a series via remote control execute the following steps a Press Start Stop b Enter the number of the first and the last series to be performed each entry Should be confirmed by the E button c Select lt REMOTE gt to enter the remote control mode The FAMOS Well Plate Microautosampler will now operate as slave of another device and can be controlled with NEXT INJ ECTION INPUT or NEXT WELL INPUT d Press Escape to return to the Ready Menu At the end of the series the message Series completed via remote control is displayed Users Manual FAMOS D931R1 3 9 The User Interface 3 8 Menus and Operating Commands This section describes all the features and possibilities of the soft keys included in the FAMOS Well Plate Microautosampler firmware in the order in which they appear on the scre
10. 1 Place the other end of the hose in a waste container on the floor All leakage of solvents and condensed water are drained through this hose if Peltier cooling option is installed Re Note Make sure that the flow path of the hoses is not obstructed in any way User s Manual FAMOS D931R1 2 9 Installation and Getting Started 2 5 3 Filling the Wash Solvent Bottle The FAMOS Well Plate Microautosampler is shipped with a 100 mL wash solvent bottle To install the wash solvent bottle a Fill the wash solvent bottle with the appropriate wash solvent Use of mobile phase in case of isocratic separations and mobile phase A without any salt or modifiers added to it in case of gradients is recommended Before using the wash solvent degas it with Helium or an ultrasonic bath or degas it on a continuous basis b Screw the bottle to the cap in the holder c Place the holder in the microautosampler as indicated in FIGURE 2 8 d Put the wash solvent tube in the wash solvent e Flush the syringe Section 2 5 4 Injection valve Wash Solvent FIGURE 2 8 Wash Solvent Bottle Waste If you use an application that requires more than 100 mL of wash solvent for a complete run use a 250 mL wash solvent bottle P N 162033 or install a longer tube with flanged end for valve fitting and place a larger bottle next to the FAMOS Well Plate Microautosampler T
11. CLICK These commands are used to allow the user to present a ERROR BEEP audible sound when a key is pressed when an error Is ALARM BUZZER observed or if an alarm Is observed TABLE 3 1 presents an overview of the default general settings and their ranges TABLE 3 1 Overview of General settings lt GENERAL gt Default Setting Range Parameter Micro Conventional Micro Conventional asom TS or 1000 aL Headspace no yes or no pressure on or off Note a 250 500 uL syringe use 500 uL buffer tubing 500 1000 uL syringe use 2000 uL buffer tubing 3 14 D931R1 User s Manual FAMOS The User Interface 3 8 4 The System Menu Item lt USAGE gt lt USAGE gt This eon function key is used to access the following settings This entry is used to enter a code for protection of methods A six digit code 000000 999999 can be selected and is erased by pressing CL If a code has been defined it is not possible to enter the System Menu and the programming menus without entering the protection code Default none PROTECTION CODE This entry is used to enable disable the ability to program timebase methods The microautosampler controls other connected equipment during analysis time Timebase methods are programmed via the Methods Menu Default disabled This entry is used to enable disable the ability to program mix methods for the microautosampler Program mix methods in accessed via the Methods Menu TIMEBASE MET
12. Communication 1 7 Options 1 8 Plate Dimensions Injection Principles A 1 Full Loop Injections A 1 1 Standard Mode A 71 2 Using an Air Segment A 2 A 2 1 Standard Mode A 2 2 Using an Air Segment A 3 A 3 1 Standard Mode A 3 2 Using an Air Segment A 4 A 5 Partial Loopfill Injections uL Pick Up Injections Low Dispersion Injection Comparing the Various Sample Injection Modes Programming Charts B 1 Overview of the Operating Program Additional Programming Examples Cal C 2 C 3 C 4 C5 C 6 C 7 Overview A 1 0 uL Partial Loopfill A 3 x 1 0 uL Injection with uL Pick Up and Wash between Injections 6 8 6 10 6 11 6 13 6 13 6 13 6 14 6 15 6 15 7 1 7 1 7 1 7 2 7 2 7 2 7 2 7 3 7 3 1 4 A 1 A 1 A 1 A 10 B 7 B 1 C 7 C 1 C 2 C 4 A 1 10 Dilution followed by a 1 0 uL Partial Loopfill Injection C 6 Defining a Template and Adding a Protection Code luL injections out of 25 wells of a 384 well plate Column Switching The Sample Injector D 1 D 2 D 3 Removing the Valve from the Microautosampler Maintenance Disassembly Reassembly of the Valve C 9 C 10 C 12 D 1 D 1 D 2 D 2 User s Manual FAMOS D931R1 Table of Contents D 3 1 Disassembly of the Valve D 2 D 3 2 Reassembly of the Valve D 3 APPENDIX E Digital Inputs and Outputs E 7 E 1 Introduction E 1 E 2 Contact Closure Outputs E 2 E 2 1 P1 Connector OUTPUTS E 2 E 2 2 P4 Connector
13. Injections A 2 1 Standard Mode The switching sequence for a partial loopfill injection is schematically presented in this section The first three steps are identical to those for Full Loop injections see Section A 1 For partial loopfill injections the sample loop is filled by transporting the programmed injection volume into the sample loop FIGURE A 7 lt Syringe gt ANNAN AVANT FIGURE A 7 Partially Filling the Injection Loop with the Programmed Injection Volume The injection valve switches into the INJECT position FIGURE A 8 and the sample loop is now part of the HPLC mobile phase flow path At this point the sample is transported to the column and the analysis time starts INT AI Sample FIGURE A 8 The Injection Valve Switched into the INJECT Position The next injection sequence will start with a flush of 50 of the programmed flush volume if the next injection is from the same vial and no wash routine is programmed Otherwise it will start with a flush of the programmed flush volume If the aspiration of sample for the next injection will exceed the total volume of the sample buffer tubing the buffer tubing is emptied before the next injection The next injection will start with the programmed flush as described for full loop injections A 4 D931R1 User s Manual FAMOS Injection Principles A 2 2 Using an Air Segment An air segment can be used to reduce the amount of flus
14. Methods Method lt INJECTION gt lt WASH gt lt TIMEBASE gt Injection methods MASNOG Timebase program Mix method Wash between 4x AUX 1 ASPIRATE Series 4x AUX 2 sample lt FULL gt lt PARTIAL gt lt PICK UP gt ene 4x AUX 3 esnan Injections 4x AUX 4 air A spe 4x ISS A reagent Analysis time Analysis time Analysis time i ee DISPENSE Flush volume Flush volume 8x CODE OUT sample 7 Injections well Injections well destination END TIME Inject control Inj Volumes waste STDIL D Inject control reagent STDIL D REPEAT WAIT Flowrate Flowrate WASH L D factor L D factor Mix method Injection method Series Wash method Time base method Use calibration wells yes no calibration wells calibration interval Sample wells Destination wells When the conventional autosampler set up is selected in Reagent wells the system setting the value ranges of the system settings Labeled wells and in the methods will be adjusted to the needs of the conventional autosampler Also will the Low Dispersion mode in the injection methods Start Stop First series no longer be available Last series lt START gt lt REMOTE gt Note These parameters depend on the contents of o the methods used and or the System Priority Injection method Settings of the FAMOS Wash method Note optional Soft function keys Time base method Note only if Feeder option is installed Well number PRIORITY Interrupt Not used
15. Packings Netherlands BV 80 Carolina Street Abberdaan 114 San Francisco CA 94103 1046 AA Amsterdam USA The Netherlands Phone 415 552 1855 Phone 31 20 683 9 68 Fax 415 552 1858 Fax 31 20 685 3452 vi D931R1 User s Manual FAMOS Instructions for Returning Instruments Before you return any item for repair please contact the nearest LC Packings office or its local distributor for instructions and obtain a return authorization number Pack the equipment carefully preferably in its original carton and ship it to the LC Packings Service Department using the appropriate address North America Europe and Asia LC Packings U S A Inc LC Packings Netherlands BV 80 Carolina Street Abberdaan 114 San Francisco CA 94103 1046 AA Amsterdam USA The Netherlands Phone 415 552 1855 Phone 31 20 683 9768 Fax 415 552 1858 Fax 31 20 685 3452 IMPORTANT 1 Make certain that the return authorization number is indicated on the address label of the package so that we can properly track and account for your system 2 Please include the following a Company letterhead with the following information Your Name Complete Mailing Address Telephone Number fax number and e mail address Return Authorization Number A detailed description of the problem The name of the LC Packings personnel to whom you have spoken to regarding the problem e Return Shipping Information if appropriate b Relevant chromatogra
16. Pico Series U Guard u Precolumn NanoPrecolumn Cheminert is a Registered Trademark of Valco Instruments Co Inc Windows is a Registered Trademark of Microsoft Corporation Printed in the Netherlands D931R1 Table of Contents WARRANTY INSTRUCTIONS FOR RETURNING INSTRUMENTS WARNINGS AND SAFETY PRECAUTIONS CE DECLARATION OF CHAPTER 1 CHAPTER 2 CHAPTER 3 CONFORMITY System Overview 1 1 Features and Design of the FAMOS Well Plate Microautosampler 1 2 Principle of Operation of the Microautosampler 1 3 General Design of the FAMOS Well Plate Microautosampler 1 4 Contents of this Manual Installation and Getting Started 2 1 Installation 2 1 1 Location of the FAMOS Well Plate Microautosampler in the Laboratory 2 2 Unpacking 2 3 The Standard Microautosampler Configuration 2 4 Electrical Connections 2 4 1 Inputs and Outputs 2 4 2 P4 Connector MARKERS 2 4 3 P5 Connector AUXILIARIES 2 4 4 P1 P2 P3 and P6 Connectors 2 4 5 Connector Communication 2 4 6 Power Connector 29 Connecting the Autosampler to the HPLC System 2 5 1 Preliminary Operations 2 5 2 Connecting Waste Tubing 2 5 3 Filling the Wash Solvent Bottle 2 5 4 Flushing the Syringe 2 5 5 Adjusting the Needle Height 2 6 Routine Operation of the System 2 6 1 Sample and Mobile Phase Considerations 2 6 2 Plates and Sample Handling 2 6 3 Placing Reagent Vials Transport Vials in Position The User Interface 3 1 Overview 32 Powering up the FAMOS Wel
17. This example describes the injection of 1 uL from out of 25 wells from a 384 well plate a Place the samples in well positions B5 B14 G14 G22 and M3 M8 b Enter the systems settings command from the lt GENERAL gt menu and select the 384 well plate c Program the injection method as shown in TABLE C 1 7 TABLE C 17 Programming the Injection Method Methods lt INJECTION gt 01 E lt PARTIAL gt E 25 00 E 05 0 E 1 E 1 00 E lt STD gt E Escape Escape to enter the Methods Menu program injection method number 1 to select partial loopfill injection method to define an analysis time of 25 minutes to define a flush volume of 5 0 uL to define the number of injections vial to set the injection volume to 1 00 uL to select the standard injection control to return to the Ready Menu d Program the three series as shown in TABLE C 18 TABLE C 18 Programming Series Parameters Series 01 E 01 E CL E lt ROW B gt 05 E lt ROW B gt 14 E Series 02 E 02 E CL E lt ROW G gt 14 E lt ROW G gt 22 E Series 03 E 03 E CL E lt ROW M gt 03 E lt ROW M gt 08 E Escape to enter the Series Menu to define the Series number to select the injection method number to enter lt NONE gt for wash method to define location B5 as first sample vial to define location B14 as last sample vial to enter the Series Menu to define the next Series number to select the next in
18. a field e Numeric Keys 0 to 9 used to enter values in the various programming fields e CL Clear used to clear a value in a field or replace it by NONE or AUTO e E Enter used to accept the present value or setting for an entry The entered value is checked for validity and then saved Function keys a Run control keys e Start Stop used to start or stop automatic processing and data acquisition when controlled via the output connectors or to reset the system after an error has occurred e Hold Cont used to hold or continue the analysis time The analysis time is extended by the period that Hold is active Users Manual FAMOS D931R1 3 3 The User Interface e Interrupt not used e Priority used to stop a run so that a priority sample is run before analyzing the rest of the programmed sample series Before the run is interrupted processing of the present sample will be finished As soon as the priority Sample has been analyzed the analytical run is resumed A priority sample is a series of one well with an injection method a wash method and a time base method defined in a template this run is possible only if the correct settings are entered in the System Menu b Programming keys e Series used to enter the Series Menu in which series can be defined for an analytical run e Methods used to enter the Methods Menu in which methods can be programmed for use in an analytical run e Menu th
19. diagram of the FAMOS Well Plate Microautosampler is presented in FIGURE 1 1 Syringe Valve ha i al Buffer tubing Te ZN w __ ep Injection ET Valve L Needle pair O e e e e f i Syringe FIGURE 1 1 Schematic Diagram of the FAMOS Well Plate Microautosampler The FAMOS Well Plate Microautosampler includes the following components e Injection Valve includes the loop to place the sample in the mobile phase e Needle Pair used to pressurize the sample vial and withdraw sample e Buffer Tubing provided to eliminate contamination of the syringe especially useful with small samples e Syringe used to aspirate the sample from a well into the sample loop To prevent contamination of the syringe the autosampler is equipped with buffer tubing between the syringe and the injection valve e Wash solvent is used to remove the sample from the buffer tubing and sample needle and to rinse the buffer tubing and sample needle The FAMOS Well Plate Microautosampler provides three different methods of injection for an analytical run e Full loop The sample loop is co
20. gt 3 20 User s Manual FAMOS D931R1 Index Timebase method 3 7 3 20 lt TIME DISPLAY gt 3 14 Timed Outputs E 4 Transport liquid 1 2 A 3 Vials first last 3 16 4 7 TTL Inputs E 5 TTL Outputs E 4 Tray Cooling Testing 5 6 Troubleshooting 6 10 lt TYPE OF PLATE gt 3 16 U lt UNLOAD gt 3 23 Unpacking 2 2 Usage Menu 3 15 lt USER PROGRAM gt 3 15 3 22 User Interface 3 1 User Program 3 7 enable disable 3 15 lt UTILS gt 3 11 V Valco Valve D 1 lt VALVES gt Timebase Method 3 20 User Program 3 23 Voltage Selector 6 8 W lt WAIT gt Mix Method 3 22 User Program 3 24 lt WAIT IN gt 3 24 Warnings ix Warranty v lt WASH gt 2 11 Mix Method 3 22 Ready Menu 3 10 User Program 3 23 Wash Method 3 19 Wash Menu 3 19 Wash Method 3 7 Wash Solvent Bottle 2 10 lt WASTE gt 3 23 Waste Tubing 2 9 lt WELL MARKER PULSE LENGTH gt 3 16 lt WELL PROCESSING METHOD gt 3 16 Wells see Plates Y lt YES gt 3 25 l 4 D931R1 User s Manual FAMOS
21. limitation any warranty of merchantability or fitness for a particular purpose Your exclusive remedies and LC Packings s sole liability for any non conformity or defect in the products and such software will be those expressed herein Under no circumstances will LC Packings s liability arising from the performance or failure to perform of any product or software in contract in tort including negligence or otherwise exceed the purchase price of the product and software In no event will LC Packings be liable in contract in tort including negligence or otherwise for special incidental consequential or analogous damages including without limitation damages resulting from loss of use loss of profits loss of business or loss of goodwill even if LC Packings has been advised of the possibility of such damages This warranty comprises the entire warranty between LC Packings and the customer It overrides any warranty related language that may appear in the customer purchase order or other documentation provided by the customer This warranty shall be governed by and construed and enforced in accordance with the laws of the Netherlands It is non transferable and shall run to the benefit of the original purchaser only Any change alteration or amendment to this warranty is not valid unless it has been approved in writing by an officer of LC Packings Warrantor North America Warrantor Europe and Asia LC Packings U S A Inc LC
22. location C3 as last sample vial Escape to return to the Ready M enu e Run the series by entering the parameters indicated in TABLE 4 8 and pressing lt START gt TABLE 4 8 Running the Series Start Stop to start the FAMOS Well Plate Autosampler 01 E to start at series 01 02 E to stop after series 02 lt START gt to start execution of the series At the end of the defined analysis time the Ready M enu will appear again to indicate that the FAMOS Well Plate Microautosampler is ready for the following next run 4 6 D931R1 Users Manual FAMOS Programming Examples 4 4 4 Example 3 luL injection out of a 5 uL sample in vial Al using the transport liquid uL pick up This example describes an application in which a single injection is made from a vial position Al using transport liquid for pickup The uL pick up routine allows for an injection to be made from small sample volumes with essentially no Sample loss To perform this operation a Place a 250 uL tapered sample vial in vial position Al and one of the 10 mL Capped vials in transport vial position 1 leftmost position in the rack as Shown in FIGURE 4 2 the transport liquid should have a lower strength than the mobile phase Wash Position Transport Solvent Positions ZIJIILIIIK ZII IIIII L TEE YE FE 0000000 FIGURE 4 2 48 Well Plate Holder and Location of Transport Liquid Sample Vials b Install a 10uL sample loop c Set the loop vo
23. lt WASH gt to connect the syringe to wash solvent bottle lt WASTE gt to connect the syringe to the waste tubing This command is used to control the movements of the syringe lt LOAD gt the syringe with the programmed volume lt SYR gt lt UNLOAD the syringe with the programmed volume lt HOME gt the volume previously aspirated will be dispensed to the last programmed position and the syringe will be initialized again This command is used 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 lt WASH gt eS Note The wash position may be contaminated with the contents of the buffer tubing which may generate cross contamination To prevent contamination of the wash position program a dispense to waste action before programming a wash action This command is used to program positions of high pressure valves ISS injector valve SSV The injector valve has two positions lt INJ ECT gt and lt LOAD gt The ISS optional valve has positions indicated by 1 6 left and 1 2 right A lt VALVES gt Users Manual FAMOS D931R1 3 23 The User Interface This command is used to program a pause max 9 hours lt WAIT gt 59 minutes 59 seconds E Note During the pause the needle will move to home position if the previous step is an aspirate or dispe
24. more specific information on IO connections 3 16 D931R1 User s Manual FAMOS The User Interface TABLE 3 2 Overview of IO settings IO Parameter Default Range Inject marker pulse length 1 0 s 0 1 2 0 Well marker pulse length 1 0 s 0 1 2 0 Labeled well marker pulse length 1 0 s 0 1 2 0 Input edge next injection falling falling or rising Input edge next well falling falling or rising Freeze input active low low or high Reset outputs after last series no yes or no 3 8 7 The System Menu Item lt CLOCK gt This entry is used to switch the system clock on or off If the ON selection is made you can set date yy mm dd and time hh mm This date and time will be displayed in the Ready Menu lt CLOCK gt 3 8 8 The System Menu Item lt COMM gt This entry to define a device identifier for communication with other equipment e g a PC An identifier betw een 20 and 29 can be selected for the FAMOS Well Plate Microautosampler default setting Is 21 lt COMM gt IES Note When controlling the FAMOS Well Plate Microautosampler by UltiChrom make certain that the same setting is used in the hardware description 3 8 9 Methods Menu When entering the Methods Menu by pressing the Method key the user can program various types of methods It is possible to define up to e 24 Separate Injection methods e 5 Wash methods e 5 Timebase methods e 9 Mix methods e 1 User program It is possible to
25. of the customer Software Warranty If at any time during the period ending ninety 90 days after delivery of any product to you you report and document any error in any software provided with such product and developed by LC Packings or any failure of any such software substantially to conform to LC Packings software description that limits or prevents use of the software by you we will use reasonable efforts to correct any such error or failure will replace such software or will terminate your license to use the software and refund the price of the related product In connection with any such termination and refund you will return the related product to LC Packings upon request The warranty will apply only to those portions of the software that were developed by LC Packings and that incorporated all program corrections and modifications if any delivered to you It will not apply to any error or failure due to machine error or to the misuse by or negligence of any person or entity other than LC Packings or to any software which is modified by any person or entity other than LC Packings User s Manual FAMOS D931R1 v Warranty Liability Under no circumstances shall LC Packings be liable for damage to persons or property This warranty is the only warranty given by LC Packings with respect to products and software provided with the products and is given in lieu of all other warranties express or implied including without
26. program a combination of methods and save them in a template The available parameters and options in a method or template may vary according the settings made in the System Menu An overview of the Methods Menu Is presented in FIGURE 3 5 Users Manual FAMOS D931R1 3 17 The User Interface Methods lt INJ ECTION gt Injection methods e Wash volume lt FULL gt lt PARTIAL gt Analysis time Flush volume Injections w ell Analysis time Flush volume Inj Volumes Inject control c STD L D Flowrate L D factor STD L D Flowrate L D factor Notes Injections w ell Inject control lt WASH gt lt TIMEBASE gt b Timebase program 4x AUX 1 4x AUX 2 4x AUX 3 4x AUX 4 4x ISS A lt MIX gt b Mix method e ASPIRATE sample destination air reagent DISPENSE lt PICK UP gt 4 x ISS B 8x CODE OUT END TIME e Inj volumes sample destination waste reagent REPEAT WAIT WASH a lt Template gt and lt User Program gt disabled in System Menu b When enabled in System Menu only c In micro mode only FIGURE 3 5 The Methods Menu on the System Menu eS Note The options provided by the Methods Menu depend on the settings made 3 8 10 The Method Menu Item lt INJ ECTION gt lt INJ ECTION gt lt FULL LOOP gt lt PARTIAL LOOPFILL gt lt uL PICK UP gt ANALYSIS TIME FLUSH VOLUME This soft function key is
27. solvent tubing A syringe volume of wash solvent is aspirated from the wash solvent bottle and the wash solvent tube is filled When the key Is selected it is redefined to lt SYR HOME gt which is used to dispense the syringe contents to syringe waste and to move the syringe to the standard operating position again lt SYR END gt D931R1 User s Manual FAMOS The User Interface lt UTILS gt This key is used to enter the Utilities Menu eS Note If a method protection code is enabled in the System Menu the code must be entered to access the Utilities Menu This key is used to copy a method select the method type mix Injection timebase wash the number of the method to be copied and enter the number for the destination method An existing method stored under that number will be overwritten lt COPY gt This key is used to erase a method Template Methods User Program If Template and User Program are disabled in the System Menu the soft function keys for erasing a standard Method mix injection wash timebase appear It is not possible to erase the user program if the protection code for the user program is enabled in the System Menu lt ERASE gt The FAMOS Well Plate Microautosampler keeps a log of system relevant events lt EVENTS gt records error messages that have been generated and keeps count of actions of valves and syringe movements lt COUNT gt The message Lifetime of syring
28. steps for the user program and all nine mix methods cannot exceed 240 The user program can be protected by a special user program protection code System Settings Usage Menu If no user program has been programmed yet end of user program is displayed Otherwise the first line of the programmed method appears The following entries are presented for the user program Press to edit an existing step or a new step for a new mix method Press to insert a new step in an existing method before the displayed step Press to delete the displayed step The edit and insert menus offer the following soft function keys Removes a programmed volume from sample well ambient air destination vial wash or one of the reagent vials into the buffer tubing The speed and height of the syringe can be entered TABLE 3 3 The maximum volume that can be aspirated is the total volume of the syringe 3 22 D931R1 User s Manual FAMOS The User Interface Delivers a programmed volume from the buffer tubing into the sample well waste destination vial wash or one of the reagent vials Speed and height of syringe can lt DISPENSE gt be entered TABLE 3 3 It is not possible to dispense a larger volume than the total volume aspirated in previous actions This command is used to control the connections of the syringe to one of its three tubes lt NEEDLE gt to connect the syringe to the sample needle lt SYR VALVE gt
29. test It includes basic system commands e g washing and plate handling a number of utilities e g copying a method and commands to establish communication with other systems e System Menu this menu is used to select settings that are changed infrequently e g the loop volume the type of plate the system clock if the alarm should be set etc In some cases these settings may have been made at the factory to suit particular wishes of the user in this instance it will not be necessary to make changes It is recommended that you enable only those facilities that you will actually need this will ensure that menus are aS concise as possible e Methods Menu this menu is used to program a method to be used in an analytical run and to assign a number to it Typical commands include the analysis time the wash volume and wash parameters e Series Menu this menu is used to define a series and to assign a number and a method to it for the analytical run If MENU or MN is displayed in the top right hand corner of the screen you can press the Menu key on the keypad to display more possibilities offered by the menu An explanation of all keys on the keypad Is presented in Section 3 3 Users Manual FAMOS D931R1 3 5 The User Interface 3 5 General Approach to Entering and Executing a Program After you have determined the type of analytical run that you want to perform the best way to generate the order of operation for t
30. the buffer tubing the syringe dispenser aspirates transport liquid from the transport vial to fill the sample line with transport liquid and remove wash solvent FIGURE A 11 Co Syringe LEER 3 Column o gt AAAA AAAA AAAA Transport liquid FIGURE A 11 The Syringe Dispenser Aspirating Transport Liquid to Fill Sample Line The needle moves from the transport vial to the sample well FIGURE A 12 The injection valve is switched to the LOAD position A 6 D931R1 User s Manual FAMOS Injection Principles lt A Syringe CELE Column gt TELEETLOL LOLI IS SALLI LALALALA Sample Transport liquid FIGURE A 12 The Injection Valve is Switched into LOAD Position The programmed injection volume is aspirated from the sample well FIGURE A 13 SJ Syringe gt i AN AANA ANIAN H Sample Transport liquid FIGURE A 13 Aspirating the Programmed Injection Volume The sample needle moves back to the transport vial FIGURE A 14 The sample is quantitatively transported into the loop with transport liquid mobile phase from the transport vial Syringe Sample Transport liquid FIGURE A 14 Transporting Sample into the Injection Loop Aspirating the Programmed Injection Volume The injection valve is switched to INJECT FIGURE A 15 The sample loop is now part of the HPLC mobile phase flow path and sample is transported to the
31. used to access commands to program a method that defines injection methods for full loop partial Loop and uL pick up operation Up to 24 methods can be programmed Select the desired injection method and then enter a number to define the injection method to be programmed If the selected method is locked because of changes in the settings System M enu after programming the method the word LOCK is displayed The method can be unlocked by programming valid values in the method itself or by restoring the values in the System Menu This entry defines the time betw een switching the injection valve to inject and the start of processing the next sample This entry defines the amount of sample taken from a vial before the loop is filled with sample Default value 5 0 uL combined with an air segment IE Note Flush volumes of less than twice the volume of the needle and tubing will result in decreased performance 3 18 D931R1 User s Manual FAMOS The User Interface NUMBER OF This entry defines the number of injections up to 9 per INJECTIONS PER Vial VIAL Indicates the injection volume for each injection from each vial The maximum programmable injection volume IS partial loopfill 50 of the programmed loop volume uL pick up injection volume loop volume 3 x needle IN ECTION VOLUME VOlume 2 full loop This parameter is not programmable and is equal to the loop volume It requires more sample to fill th
32. valve is not in a valid position Contact N e ERROR 15 The ISS valve did not switch within 1 5 Contact seconds LC Packings TABLE 6 7 Error Codes for Syringe Dispenser Unit Probable Cause Solution ERROR 21 The syringe valve did not switch Contact ERROR 21 Te eringe ete ciinerewien s eckngs ERROR 22 The syringe did not reach home position in time Contact ERROR 23 The syringe spindle did not make the correct Contact number of rotations LC Packings ERROR 24 The spindle does not rotate Contact ERROR 24 Tresor doss noras e pucings ERROR 25 The syringe valve did not find a valid position Contact LC Packings TABLE 6 8 Error Codes for Plate and Plate Holder Probable Cause ERROR 59 Check for Plate ERROR 90 Plate home time out plate did not reach home Reinstall Plate position home error Holder ERROR 91 Plate did not reach or leave home position during Reinstall Plate ERROR 92 Plate holder missing Check reinstall a Plate Holder ERROR 93 Plate holder movement is blocked Check for blockage User s Manual FAMOS D931R1 Maintenance and Troubleshooting TABLE 6 9 Error Codes for Injection needle unit Error Code Probable Cause Solution ERROR 30 The sample needle arm did not reach or leave Contact Rs home position vertical LC Packings ERROR 31 The sample needle arm is in an invalid horizontal Contact ERROR ST pester whi moving dow Le Peekings ERROR 32 The samp
33. warranty covers products sold under the LC Products trademark If a different warranty than the above is indicated in the sales literature the warranty indicated in the sales literature will prevail If the system includes equipment supplied by LC Packings but manufactured by a third party LC Packings makes no warranty of any kind express or implied including without limitation any warranty of merchantability or fitness for a particular purpose LC Packings will make available to you to the extent permitted the warranties of the manufacturer of the relevant equipment following your timely written request If any product covered by this warranty becomes defective during the warranty period it will be repaired or replaced by LC Packings at no charge to the customer the repair replace decision is solely at the option of LC Packings All warranty requests must be received by LC Packings during the warranty period LC Packings will pay for surface transportation to the applicable LC Packings Office North America San Francisco CA Europe and Asia Amsterdam the Netherlands if the instrument proves defective within thirty 30 days from the date of shipment this does not include air freight drayage labor crating charges customs clearance charges etc The user should carefully follow the directions indicated on the Return Goods Instruction Sheet in the User s Manual After thirty days all transportation costs will be at the expense
34. 3 E to define the number of injections per well 100 E to enter the injection volume for 1st injection 100 E to enter the injection volume for 2nd injection 100 E to enter the injection volume for 3rd injection Escape to return to the Methods Menu d Program the mix method by entering the parameters indicated in TABLE C 11 C 6 D931R1 User s Manual FAMOS Additional Programming Examples TABLE C 11 Programming Mix Methods Parameters lt MIX gt 1 E to enter the Mix Menu and define Mix method no 1 lt INSERT gt to define mix method step number 1 lt ASPIRATE gt 100 E to aspirate 10 0 uL from reagent vial A Menu lt REAG A gt E lt INSERT gt to define mix method step number 2 lt DISPENSE gt 90 to dispense 9 0 uL to destination well lt DESTINATION gt lt INSERT gt to define mix method step number 3 lt WAIT gt 02 E to wait 2s lt INSERT gt to define mix method step number 4 lt DISPENSE gt 10 to dispense 1 0 uL to waste lt WASTE gt lt WAIT gt O2 E to wait 2s lt INSERT gt to define mix method step number 6 lt ASPIRATE gt 20 E to aspirate 2 0 uL air lt AIR gt lt WAIT gt O2 E to wait 2s lt INSERT gt to define mix method step number 8 lt ASPIRATE gt 20 E to aspirate 2 0 uL sample lt SAMPLE gt lt WAIT gt O2 E to wait 2s lt INSERT gt to define mix method step number 10 lt DISPENSE gt 10 E to dispense 1 0 uL to the destinati
35. 3 23 lt LOG gt 3 11 lt LOOP VOLUME gt 3 13 Loop Volume 6 7 Testing 5 6 Low Dispersion Injection A 9 M Mains Input 2 7 Mains Switch 2 7 Maintenance 6 2 lt MANUAL gt 3 12 lt MARKERS gt User Program 3 24 Markers P4 Connector 2 6 E 2 MENU 3 5 Menu Key 3 4 Methods Key 3 4 Methods Menu 3 5 3 17 lt METHODS gt lt INJECTION gt 3 18 lt MIX gt 3 21 lt TEMPLATE gt 3 25 lt TIMEBASE gt 3 20 lt USER PROGRAM gt 3 22 lt WASH gt 3 19 Methods Menu 3 5 3 17 Method Types 3 7 lt MICRO gt 3 13 lt MIX gt 3 21 Mix Menu 3 21 Mix Method 3 7 enable disable 3 5 lt MIX METHODS gt 3 15 Mobile Phase Considerations 2 12 MN 3 5 N Needle Height Adjusting 2 11 Optional 6 5 Prepuncturing 6 4 6 6 Replacing 6 4 Sample 6 4 lt NEEDLE gt 3 23 lt NEEDLE HEIGHT 3 14 l 2 D931R1 User s Manual FAMOS lt NEEDLE TUBING gt 3 13 lt NO gt 3 25 lt NUMBER OF INJECTIONS PER VIAL gt 3 19 Numeric Keys 3 3 O Outputs 2 5 Contact Closure E 2 Timed E 4 TTL E 4 P lt PANIC gt 3 12 lt PARTIAL LOOPFILL gt 1 2 3 18 Partial Loopfill A 4 Standard Mode A 4 Air Segmentation A 5 Peltier Cooling 1 1 3 12 Plates Handling 2 12 Menu 3 16 lt PLATES gt 2 13 3 10 3 16 Plate Holder Installation 2 8 lt PLATE HOME gt 2 13 3 10 Power Connector 2 7 Powering Up 3 2 Preparations for Use 2 5 Prepuncturing Needle 6 4 Replacing 6 6 Principle of Operation 1 2 Priority key 3 4 Priority S
36. 3 and should not be changed COMMUNICATION Default Settings FIGURE 2 3 Default DIP Switch Settings Note Older versions of the FAMOS Well Plate Microautosampler are equipped with only one 25 pin connector instead of two 9 pin connectors If the autosampler includes the DIP switches they should be set to the default settings presented and not be changed 2 6 D931R1 User s Manual FAMOS Installation and Getting Started 2 4 6 Power Connector The FAMOS Well Plate Microautosampler is equipped with a power supply for input voltages from 110 to 120 VAC and 220 to 240 VAC Changing of the setting is only required if the voltage indicated by the voltage selector FIGURE 2 4 on the back panel does not match with the mains voltage FIGURE 2 4 indicates the setting for 220 240 V operation The voltage indication plate can be removed with a small screwdriver A AOQcL OLL AOSz 3G S31gISN4 S3G D3AV LNSIWANOINN YSAOTAING S3SN4 AOSZ HLIM AINO 3sn 220 240 VV SAO TMS SASNS AOGe HALIM INO Bon V 220 240 VV O tage Indication FIGURE 2 4 Power Entry Module The power cord should be inserted in the socket directly below the Main Power switch e Check whether local voltage matches voltage indicated on back panel of the FAMOS Well Plate Microautosampler e Connect the power cord to the FAMOS Well Plate Microautosampler item 5 FIGURE 2 2 e Switch the FAMOS Well Plate on by using the switch at the back pane
37. 440 mm 17 2 in WxDxH 23 kg 50 7 Ib 7 1 5 Electrical Power 115 VAC 15 20 50 Hz 60 Hz 250 VA Requirements 230 VAC 15 20 50 Hz 60 Hz 250 VA 115 VAC two 5 0 AT fuses 4 x a UL CSA Fuses 230 VAC two 2 5 AT fuses 5 x 20 mm IEC 127 All fuses UL listed and CSA certified 7 2 D931R1 User s Manual FAMOS Specifications 7 1 6 Communication Inject marker Well marker Labeled well marker Stop I O 4 Auxillary outputs 2 Programmable outputs Alarm output 4 Bit timebase Next injection input Next well input Inputs Freeze input Stop I O 4 Programmable inputs Communication 7 1 7 Options Outputs Built in Peltier cooling processing unit Programmable Range 4 C 40 C Cooling capacity Ambient 20 C measured on cooling plate plate holder removed Sample cooling Note The temperature of the sample inside the sample vial may be slightly different than the temperature set via the command due to a variety of effects e g the heat transfer characteristics of the vial walls If it is necessary to precisely set the temperature inside the vial we recommend that you determine the temperature inside the vial at various temperature setpoints and set the cooling option so that the desired internal temperature is attained THERMOS column oven combined with the FAMOS Well Column oven Plate Microautosampler The injection valve and optional high pressure valve fit into the co
38. Assembly To assemble the PEEK sleeve fitting using the supplied stainless steel union a Slide the fingertight nut and the ferrule onto the sleeve as shown in FIGURE G 1 b Insert this assembly into the union that is provided screwing the nut in two or three turns by hand c Push the sleeve all the way forward into the union so that the sleeve seats firmly This is essential for a proper zero dead volume connection FIGURE G 2 Attaching a PEEK Sleeve connector to a NANO column d Manually turn the nut into the union until it is finger tight Carefully turn the nut 1 4 turn 90 past the point where the ferrule starts to grab the sleeve FIGURE G 2 Use a 4 wrench to retain the union e Remove the pre assembled sleeve fitting and inspect it The ferrule should be firmly attached to the tubing i e you should not be able to move it laterally along the tubing axis If the ferrule is loose reinstall the tubing in the fitting in the union and tighten it another 1 8 of a turn past finger tight f Push the NANO column capillary all the way forward into the pre assembled sleeve so that the sleeve and column seat firmly This is essential for a proper zero dead volume connection Ie Note It is essential that the sleeve and the NANO column capillary are inserted all the way forward into the union for a proper zero dead volume connection g Manually turn the nut into the union until it is finger tight Carefully turn th
39. D931R1 User s Manual FAMOS Installation and Getting Started IIe Note The instrument has been flushed with isopropanol before shipment from the factory Make sure that the mobile phase of your HPLC system is miscible with isopropanol or start up with an intermediate solvent as mobile phase disconnect the HPLC column when flushing IIe Note It is very important that the contents of the sample loop are injected in back flush mode onto the column therefore do not exchange column and pump connections at the injection valve e Power up the system The display will indicate that the self test and initialization have been executed After completion of this procedure the screen shown in FIGURE 2 7 will be presented 12 04 THURSDAY JANUARY 04 01 MENU READY 2 02 lt EXCHANGE gt lt WASH gt lt SYREND gt lt UTILS gt FIGURE 2 7 The Ready Menu 2 5 2 Connecting Waste Tubing To connect the three waste tubes e Syringe waste put the end of the syringe waste tube item 15 FIGURE 2 1 in a bottle placed next to the FAMOS Well Plate Microautosampler e Drain wash position connect the hose to the drain wash connector of the FAMOS Well Plate item 10 FIGURE 2 1 place the other end of the hose in a bottle placed on the floor All liquid dispensed to waste at the back of the plate is removed through this drain e Condensed water and leakage connect the hose to the drain port of the FAMOS Well Plate item 11 FIGURE 2
40. Every Day Before operating check for any air bubbles in the fluidic lines and degas the wash solvent connections fluidics joints When using buffer solutions flush the system thoroughly after use with a solvent that contains does not contain buffers salts Every 3 months Inspect the condition of all tubing cracks nicks cuts clogging Every year LC Ie Note The frequency of the various activities described above is a good starting point As the user gains experience with the system it will be found that some activities can be done less frequently and other need to be done more frequently The frequency is dependent on a number of factors including the nature of the sample and the mobile phase 6 2 D931R1 User s Manual FAMOS Maintenance and Troubleshooting 6 3 Replacing Major Components A variety of components on the FAMOS Well Plate Microautosampler can be readily changed by the user as required to ensure that the instrument is maintained in optimal condition In some circumstances the analyst may want to change a component as an example four different needles are available see Section 6 6 2 and it is possible that the application would be best served with a silica coated needle if the sample could interact with the stainless steel needle 6 3 1 Replacing the Syringe The FAMOS Well Plate Microautosampler is supplied with a 25 uL syringe standard configuration In addition a 100 250 500 or 1000 uL syr
41. FAMOS Well Plate Microautosampler User s Manual P N 160557 Now sold under the Thermo Scientific brand Thermo SCIENTIFIC LC PACKINGS A O Company www lcpackings com D931R1 Notice The FAMOS Well Plate Microautosampler is covered by a limited warranty A copy of this warranty is included with this manual The customer is required to perform routine maintenance as described in the User s Manual on a periodic basis to keep the warranty in effect All information in this manual is subject to change without notice and does not represent a commitment on the part of LC Packings BV The material included in this manual is provided to assist users in the operation maintenance and repair of the FAMOS Well Plate Microautosampler It is assumed that the individual using this manual has sufficient training in the use of analytical instrumentation and is aware of the potential hazards including but not limited to electrical hazards chemical solvent hazards and the exposure to pressurized solvents November 2001 LC Packings BV A Dionex Company All rights reserved No part of this manual may be reproduced or transmitted in any form or by any means without the written permission of LC Packings BV The following are registered trademarks of LC Packings A Dionex Company UltiMate UltiChrom FAMOS Thermos Switchos Acurate UZ View UV Booster FluoBoost Pepmap Dumper u Fluidics Fusica Nano Series
42. FAMOS D931R1 4 3 Programming Examples 4 4 Sample Applications 4 4 1 Conventions The following typographical conventions are used Initial Capitals represents the names of the menus e g Ready Menu lt CAPTALS gt represents the soft function key e g lt INJ ECTION gt bold type represents the names of the function keys e g E 01 23 indicates that a numerical value has to be entered 4 4 2 Example 1 Injecting a Single 200 nL Sample from 1 vial This example describes an application in which a single injection is made from a Single vial in position Al a Place a sample in position Al b Program the Injection Method by entering the parameters indicated in TABLE 4 2 TABLE 4 2 Programming the Injection Method Methods to enter the Methods Menu lt INJ ECTION gt 01 E to define the number of the method lt PARTIAL gt E to select partial loop fill 25 00 E to set analysis time to 25 min 5 0 E to set the flush volume to 5uL 01 E to set the number of injections vial to 1 02 E to set injection volume to 0 2 uL lt STD gt E to select the standard injection control Escape Escape to return to the Ready M enu c Program the Series by entering the parameters indicated in TABLE 4 3 TABLE 4 3 Programming the Series Series to enter the Series M enu 01 E to define the Series number 01 E to select the injection method number CLE to enter lt NONE gt for wash method lt ROW A gt 01 E to
43. HODS MIX METHODS eS Note The microautosampler cannot analyze priority samples during a run if the mix method is enabled Default disabled This entry is used to enable disable the possibility to generate a user program If this function is enabled it is USER PROGRAM possible to enter a user program protection code 6 digits A user program is generated via the Methods Menu Default disabled Ie Note The microautosampler cannot analyze pnority samples during a run if the user program is enabled This entry is used to enable disable the possibility to LABELED WELLS program labeled wells The location of labeled wells is programmed via the Series Menu Default disabled This entry is used to enable disable the possibility to TEMPLATES program templates Templates are programmed via the Methods Menu Default disabled This entry is used to enable disable the possibility of CALIBRATION programming calibration wells The location of the WELLS calibration wells is entered in the Series Menu Default disabled ae Note We recommend that you disable as many functions in the Usage Menu as possible to make sure that other menus do not contain possibilities that are irrelevant for the type of analyses you are to perform Users Manual FAMOS D931R1 3 15 The User Interface 3 8 5 The System Menu Item lt PLATES gt lt PLATES gt This soft function key is used to access the following settings This entry is used
44. MARKERS E 2 E 2 3 P5 Connector AUXILIARIES E 3 E 3 TTL Outputs E 4 E 3 1 P2 Connector TTL OUTPUTS E 4 E 3 2 P3 Connector TIMED OUTPUTS E 4 E 4 TTL Inputs E 5 E 4 1 P6 Connector TTL Inputs E 5 APPENDIX F Programming Forms F 7 APPENDIX G Connecting the NANO Column or other Fused Silica Capillary G 7 G 1 Introduction G 1 G 2 Using PEEK Fingertights G 1 G 3 Fitting Assembly G 2 G 4 Using Long PEEK Hex Style Nuts G 3 G 5 Spare Parts Lists G 4 INDEX l 1 iv D931R1 User s Manual FAMOS Warranty LC Packings Netherlands BV warrants that the products manufactured and sold by it to be free from defects in material and workmanship for normal use and service from the date of delivery to original purchaser for a period of one 1 year from the date of shipment This limited warranty does not cover and no warranty is provided for parts that by their nature are required to be replaced periodically as a function of use of the normal operation of the system These items include without limitation HPLC columns fuses tubing detector sources pump piston seals injector rotors check valves filters any software etc In addition damage due to corrosion misuse negligence accident alteration of the system or repair by an unauthorized individual is not covered by the warranty It is understood that the performance characteristics of the instrument require that the mobile phase be degassed with He as described in the User s Manual This
45. MOS Well Plate Microautosampler as well as the inert version If you are using an inert version please refer to Appendix G which includes specific information that relates to this configuration and the appropriate part numbers for replacement parts This manual describes the FAMOS Well Plate Microautosampler and includes the following information Chapter 2 Installation and Getting Started describes how to install the FAMOS Well Plate Microautosampler Chapter 3 The User Interface describes the use of the display panel and the user interaction program to establish methods and series Chapter 4 Programming Examples presents details of a variety of methods series for that represent common applications of the microautosampler Chapter 5 Testing the Microautosampler includes a protocol that can be used to verify that the Microautosampler is operating in an acceptable manner Chapter 6 Maintenance and Troubleshooting describes a variety of maintenance procedures to optimize the performance of the microautosampler In addition it discusses how the operator can determine the cause of a difficulty in the operation of the autosampler and includes a list of spare replacement parts Chapter 7 Specifications presents the specifications of the FAMOS Well Plate Microautosampler In addition a series of appendices are provided to supply information about the injection principles a detailed chart that presents the operation program additional pr
46. Model C2 Valve b To insure that the sealing surface of the cap is not damaged rest it on its outer face If the tubing is still attached leave it suspended by the tubing c Gently pry the rotor away from the driver with your fingers or a small screwdriver d Examine the rotor sealing surface for scratches e If scratches are visible to the naked eye the rotor must be replaced e If no scratches are visible clean all parts thoroughly with an appropriate solvent Take care that no surfaces get scratched while you are cleaning the components Ie Note The most common problem in the use of the valve with HPLC is the formation of buffer crystals which are usually water soluble After cleaning it is not necessary to dry the rotor D 2 D931R1 User s Manual FAMOS The Sample Injector D 3 2 Reassembly of the Valve To reassemble the valve a 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 b Replace the cap Insert the two socket head screws and tighten them gently until both are snug CAUTION Caution Do not overtighten the screws they simply hold the assembly together and do not affect the sealing force which is automatically set as the screws close the cap against the valve body c Test the valve by pressurizing the system If the valve does not hold pressure it should be returned fo
47. NO Se ae Power ground PIN Spare Alarm output s O 15 Note Vmax 28 Voc Vac Imax 0 25 A Connector P1 OUTPUTS includes 2 programmable outputs and an alarm output Note The Alarm output will be activated whenever an error occurs A description of the error codes of the FAMOS Well Plate Microautosampler is presented in Chapter 6 E 2 2 P4 Connector MARKERS TABLE E 2 P4 Connector Pin Descriptions Normally open Inject Common Marker Normally closed Normally open Well Common Marker Normally closed 24 V DC Normally Closed edini o o a NC COM O Normally Open NO a Stop 1 0 Normally closed Vmax 28 Voc Vac Imax 0 25 A Vmax 28 Voc Vac Imax 0 25 A E 2 D931R1 User s Manual FAMOS Digital Inputs and Outputs E 2 3 P5 Connector AUXILIARIES TABLE E 3 P5 Connector Pin Descriptions Normally open Normally open AUX 1 Common Normally closed Normally closed Normally open Normally open AUX 2 Common Normally closed Normally closed Normally Closed Note ommon NC PECON a Vmax 28 Voc Vac Imax 0 25 A O Normally Open NO Vmax 28 Voc Vac Imax 0 25 A User s Manual FAMOS D931R1 E 3 Digital Inputs and Outputs E 3 TTL Outputs The following tables show the marker outputs P2 and a 4 bit time base code output P3 programmable in a time base method Both connectors are TTL level outputs FIGURE E 2
48. Only available when user program and mix method have been disabled in the System Settings Help Context and status sensitive help screen if available B 2 D931R1 User s Manual FAMOS Programming Charts Ready Menu lt EXCHANGE gt lt Menu gt lt PLATES gt lt WASH gt lt SYR END gt lt UTILS gt lt SERIAL gt lt COOL gt lt SERVICE gt Cool ON OFF lt COPY lt ERASE gt lt LOG gt lt DEFAULTALL gt eek ON OFF Exchange Perform Move syringe Copy method Erase methods Logbook All methods Communication Service mode Lift_left standard wash amp Userprog erased all settings protocol Lift_right to default Plate System System Settings autosampler lt MICRO gt lt CONVENTIONAL gt System Settings lt Menu gt lt GENERAL gt lt USAGE gt lt PLATE gt lt l0 gt lt CLOCK gt lt COMM gt Loop volume Method protection code Inj Marker pulse e Device identifier Needle tubing Use time base method Well marker pulse Syringe volume 1 Use mix method ROWS COLUMNS Lab Well marker pulse Syringe speed Use user program e Number of transport Input edge next inj Needle height Use labeled wells vial Input edge next well Skip missing wells Use templates Freeze input active Air segment Use calibration wells Reset outputs after Headspace pressure series Time base display Key click Error beep Alarm buzzer 1 only in Conventional mode Methods programming Method lt TEMPLATE gt lt METHODE
49. R 73 Current limit of the external I O exceeded Contact ERROR TS Serontling or me otomi VO exeeadot erackings ERROR 75 Error occurred during initialization the FAMOS Contact Well Plate Microautosampler cannot start LC Packings 6 12 D931R1 User s Manual FAMOS Maintenance and Troubleshooting 6 6 Spare Parts List 6 6 1 Major Items Standard Version Stainless Steel Injection Valve FAMOS Well Plate Microautosampler with Cooling option 10 40 C installed with Feeder option installed FAMOS Well Plate Microautosampler with Feeder and Cooling option 10 40 C installed Inert Version PAEK Injection Valve FAMOS Well Plate Microautosampler inert version with Cooling option 10 40 C installed pom rn n O 161172 coat with Feeder option installed FAMOS Well Plate Microautosampler inert version with Feeder and Cooling option 10 40 C installed 6 6 2 Needle Assembly Standard Version Stainless Steel Injection Valve Micro Mode Fused silica injection sample needle 100 um I D pre assembled 160116 2 4 uL Fused silica fraction injection needle 75 um I D extra long 160117 i version 10 3 uL P Standard Version Stainless Steel Injection Valve Conventional Mode Standard conventional stainless steel sample needle supplied with TEFZEL tubing 15uL Inert Version PAEK Injection Valve Micro Mode Fused silica injection sample needle 100 um I D pre assem
50. S Well Plate Microautosampler 25 100 250 500 and 1000 uL TABLE 6 4 Injection Ranges Injection Mode Injection Volume Range Maximal Injection Volume loop volume injection volume fixed Full loop 10 uL by loop size Partial loopfill 0 01 5 uL 50 of loop volume l loop volume 3 x needle tubing uL pick up 0 01 1 4 uL volume 2 The characteristics of each injection mode is described below e Full loop injection gives maximum reproducibility RSD lt 0 4 e Partial loopfill gives maximum accuracy depends on syringe accuracy and reproducibility better than 0 6 RSD for injection volumes gt 1 uL e pl Pick up offers zero sample loss maximum accuracy same as partial loopfill and RSD better than 1 5 for injection volumes 1 WL A discussion of the appropriate modes for three common scenarios is presented below B Injection Volumes Smaller than 1 pL Partial loopfill For maximum reproducibility and accuracy a 25 uL syringe and a 1 uL sample loop should be used pL Pick up For optimum accuracy and reproducibility a 25 uL syringe and a 10 uL sample loop should be used The sample plug is transported into the loop preceded by a plug of transport liquid that has a volume of 2 5 times the programmed needle tubing volume User s Manual FAMOS D931R1 6 7 Maintenance and Troubleshooting C Injection Volumes up to Twice the Standard With a 100 uL syringe a stainless steel needle with t
51. TION CAUTION 2 4 Electrical Connections 2 4 1 Inputs and Outputs The FAMOS Well Plate Microautosampler has six standard I O connectors P1 P6 five OUTPUT connectors and one INPUT connector The Communication connector is a standard RS 232 communication interface connector The different configurations of the I O connectors are described in Appendix E All electrical connections are made on the rear panel of the autosampler FIGURE 2 2 COMMUNICATION 1B ie 9 S 1 Serial Communication Connectors RS232 2 CE mark 3 I O connectors P1 P3 4 I O connectors P4 P6 5 Power Entry Module with Main Switch 6 Fuses and voltage selector 7 Type label 8 Fan if plate cooling is installed 9 Ventilation holes FIGURE 2 2 Rear Panel of the FAMOS Well Plate Microautosampler Caution Avoid touching the electrical contacts on the terminal strips Electrostatic discharges could damage internal components Caution The manufacturer will not accept any liability for damages directly or indirectly caused by connecting the FAMOS Well Plate Microautosampler to instruments which do not meet relevant safety standards User s Manual FAMOS D931R1 2 5 Installation and Getting Started The electrical connections that are required depend on the nature of the instrumentation and the desired application In this section we describe a standard installation with other LC Packings instrumentation If the system
52. TION 1 2 4 ISS A POSITION 6 1 4 ISS A POSITION 1 2 1 SSV PORT 2 SSV PORT 3 SSV PORT 4 SSV PORT 5 SSV PORT 6 SSV PORT 4 CODE OUT 5 CODE OUT 6 CODE OUT 7 CODE OUT 8 CODE OUT END OF TIMED EVENTS F 6 D931R1 User s Manual FAMOS Programming Forms Mix Method Method number Line Action Value Position Speed Height Line Action Value Position Speed Height 1 41 2 42 3 43 4 44 5 45 6 46 7 47 8 48 9 49 10 50 11 51 12 52 13 53 14 54 15 55 16 56 17 57 18 58 19 59 20 60 21 61 22 62 23 63 24 64 25 65 26 66 27 67 28 68 29 69 30 70 31 71 32 72 33 73 34 74 35 75 36 76 37 77 38 78 39 79 40 80 Comments User s Manual FAMOS D931R1 F 7 Programming Forms User Program Line Action Value Position Speed Height Line Action Value Position Speed Height 1 41 2 42 3 43 4 44 5 45 6 46 7 47 8 48 9 49 10 50 11 51 12 52 13 53 14 54 15 55 16 56 17 57 18 58 19 59 20 60 21 61 22 62 23 63 24 64 25 65 26 66 27 67 28 68 29 69 30 70 31 71 32 72 33 73 34 74 35 75 36 76 37 77 38 78 39 79 40 80 Comments F 8 D931R1 User s Manual FAMOS Programming Forms User Information Name of user Company Department Address Telephone Telefax FAMOS Well Plate Microautosampler Information Serial number Firmware version Purchase date Installed options Local dealer Address Telepho
53. These parameters depend on the contents of Start Stop First series the methods used and or the System Settings Last series of the FAMOS lt START gt lt REMOTE gt Note optional Soft function keys Note only if Feeder option is installed Priority Template Injection method Vial number Wash method PRIORITY Timebase method Only available when user program and mix method have been Vial number disabled in the System Settings Interrupt Not used Help Context and status sensitive help screen if available User s Manual FAMOS D931R1 B 3 Programming Charts This page intentionally left blank B 4 D931R1 User s Manual FAMOS APPENDIX C Additional Programming Examples C 1 Overview The commands described in Chapter 3 can be used by the analyst to generate a broad range of programs for the operation of the FAMOS Well Plate Microautosampler These commands allow the user to configure the unit to meet the specific needs of the laboratory A series of blank forms are provided to assist the analyst in programming Appendix F Chapter 4 describes a number of programs that the user can prepare for activities that are commonly performed with the microautosampler In this appendix we describe a few programs that are important but used less frequently In addition to the direct use of these programs they can be used to provide an understanding of the logical processes that are used
54. Y N rr ee ee a ee ee NI N A O Comments User s Manual FAMOS D931R1 F 3 Programming Forms Injection Methods i Vol Well TPP PPE PEE p tT tt P te tT a pt tt tt tt ft Injection Type O O e N OIl CO NI OIl 0O A OINI C Oo 3 O o 2 a m SS a S a as a n a as S SS a a a as S S a as S a E E a fu artia c pick u a D 5 mb T n A D931R1 User s Manual FAMOS Programming Forms Wash Methods Wash method Wash between Comments number _linjections _ wells _lseries _linjections _ wells series _ linjections wells _ series _ injections 3 wells _ series _ injections 5 _ wells series Comments User s Manual FAMOS D931R1 F 5 Programming Forms Timebase Methods Method number Action At Time Action At Time AUX 1 1 AUX 1 ON 1 AUX 1 OFF 2 AUX 1 ON 2 AUX 1 OFF 3 AUX 1 ON 3 AUX 1 OFF 4 AUX 1 ON 4 AUX 1 OFF auxzor axan zawo awn awor aaux20n aao 1 AUX 3 ON 1 AUX 3 OFF 2 AUX 3 ON 2 AUX 3 OFF 3 AUX 3 ON 3 AUX 3 OFF 4 AUX 3 ON 4 AUX 3 OFF 1 AUX 4 ON 1 AUX 4 OFF 2 AUX 4 ON 3 AUX 4 OFF 3 AUX 4 ON 3 AUX 4 OFF 4 AUX 4 ON 4 AUX 4 OFF ISS A AUX 3 AUX 4 END Comments 1 ISS A POSITION 6 1 1 ISS A POSITION 1 2 2 ISS A POSITION 6 1 2 ISS A POSITION 1 2 3 ISS A POSITION 6 1 3 ISS A POSI
55. ained between 20 and 80 relative humidity e f flammable or toxic solvents are to be used a suitable ventilation system should be provided e The use of open flames in the laboratory should be prohibited e Corrosive vapors or dust should not be present as these materials can adversely affect the long term performance of the system The microautosampler requires approximately 280 mm 11 2 of linear bench space The lab bench should be capable of supporting 100 kg 225 lb The power consumption of the FAMOS Well Plate Microautosampler is 250 VA User s Manual FAMOS D931R1 2 1 Installation and Getting Started DANGER CAUTION Danger The FAMOS Well Plate Microautosampler must be connected to a power source that is connected to a true ground In addition all other components of the system e g the HPLC pump the detector should be connected to the same ground Caution Do not install the FAMOS Well Plate Microautosampler in areas subject to shock dust or in direct sunlight Do not place it near a source of heat this is especially important if the tray cooling option is installed 2 2 Unpacking When the FAMOS Well Plate Microautosampler is received carefully unpack the unit and verify receipt of all components according to the packing list some components include sub packing lists It is recommended that all packing materials be saved in the event that it is necessary to return any item to the factory
56. al Reagent B O Vial Reagent C O Vial Reagent D Only if the use of labeled wells has been enabled in the System Menu O Labeled well no 1 O Labeled well no 2 O Labeled well no 3 O Labeled well no 4 marked questions are always asked in series O marked questions depend on the used methods and the settings entered in the System Menu 3 26 D931R1 User s Manual FAMOS The User Interface After you have entered the required settings in the System Menu and after you have programmed methods to be used for an analytical run you can press Series to enter the Series Menu TABLE 3 4 gives an overview of the items you should to define for the Series e With Templates if you are going to execute an analytical run by way of a template you will only be asked to enter the template number and to indicate the location of the first sample well and the last sample well e Without Templates if you are going to execute an analytical run without using a template you will be asked to enter an injection method number and a wash method number and you will have to indicate the location of the first Sample well and last sample well If you have enabled use of calibration wells in the System Menu Usage Menu you should define whether you will use calibration wells and indicate the location of the first and last calibration well and indicate the number of wells between calibration wells FIGURE 3 6 However if you have for example enable
57. al attention 11 Dispose of all waste mobile phase in an environmentally safe manner that is consistent with all local regulations Do not allow flammable and or toxic solvents to accumulate Follow a regulated approved waste disposal program Never dispose of flammable and or toxic solvents through the municipal sewage system 12 PEEK tubing is used in a variety of locations While this polymer has superb chemical resistance to most organic solvents it tends to swell when it is contact with CHCls DMSO and THF In addition it is attacked by concentrated acids such as Sulfuric Acid and Nitric Acid swelling or attack by acid is not a problem with short flushing procedures Do not use PEEK tubing that is stressed bent or has a kink 13 Wear protective eye goggles when handling fused silica tubing i e installation cutting etc 14 If a buffer is used as a part of the mobile phase flush the system with several volumes of a methanol water 50 50 before it is shut down This will prevent salt buildup inside the unit 15 Do not use the FAMOS Well Plate Microautosampler in ways other than those indicated in the instructions given in this manual 16 The following symbols are used on the FAMOS Well Plate Microautosampler WARNING This indicates that care should be taken to prevent personal injury or damage to parts of the FAMOS SHARP NEEDLE Well PI t AND MOVING PARTS e ate KEEP HANDS CLEAR This sticker with yellow background
58. ample 3 4 lt PROG OUT gt 3 24 Programming Charts B 1 Examples 4 1 C 1 Forms F1 lt PROTECTION CODE gt 3 15 R Ready Menu 3 2 3 5 3 10 Reagent Vials placing in position 2 13 Rear Panel 2 5 lt REMOTE gt 3 9 Remote Control 3 9 lt REPEAT gt Mix Method 3 22 Replacing Needle Assembly 6 4 Prepuncturing Needle 6 6 Syringe 6 3 Syringe Tip 6 4 Returning Instruments vii S Index Safety Precautions ix Sample Considerations 2 12 Handling 2 12 Injector D 1 Injection Modes Comparison A 10 Sample loop 6 7 Sample Needle Replacing 6 4 lt SCALE FACTOR gt 3 13 lt SERIAL gt 3 12 Series Key 3 4 Series Menu 3 5 3 26 without templates 3 26 with templates 3 26 lt SERVICE gt 3 12 Silica coated Needle 6 4 lt SKIP MISSING VIALS gt 3 14 Soft Function Keys 3 3 Spare Parts 6 13 Specifications 7 1 lt SSV gt Ready Menu 3 12 Timebase Program 3 24 Standard System Configuration 2 3 6 7 Start Stop key 3 3 lt SYR gt 3 23 lt SYR END gt 3 10 lt SYR HOME gt 3 10 Syringe replacing 6 3 types of 6 4 waste 2 9 Accuracy Checking 5 5 Syringe Speed 3 25 lt SYR_ VALVE gt 3 23 lt SYRINGE SPEED gt 3 13 Syringe Tip Replacing 6 4 lt SYRINGE VOLUME gt 3 13 Syringe Waste Tubing 2 9 System Key 3 4 System Menu 3 5 3 13 System Settings 4 2 T Temperature Control 3 12 lt TEMPLATE gt 3 15 Template Definition 3 7 Create 3 25 Enable disable 3 15 Menu 3 25 Testing 5 1 Test Protocol 5 2 5 3 lt TIMEBASE
59. and STOP I O input can be used to control the FAMOS Well Plate by other devices The four inputs INPUT 1 to 4 can only be used in the user program e g to control the sequence of the steps in this method A connection diagram is shown in FIGURE E 3 Active high Active low Autosampler Autosampler VCC VCC Input Input GND GND FIGURE E 3 TTL Input E 4 1 P6 Connector TTL Inputs TABLE E 6 P6Connector Pin Descriptions Next injection input This input will start the next injection sequence when the FAMOS Well Plate Microautosampler is started in remote control When the injection sequence is finished the FAMOS Well Plate Microautosampler will wait for the next input From the Ready Menu a NEXT INJECTION INPUT will start the last programmed series In this case the FAMOS Well Plate Microautosampler will not wait for the NEXT INJECTION INPUT before continuing with the next injection The FAMOS Well Plate Microautosampler will execute the complete RUN as if it was started with the Start Stop key Next well input With this input the FAMOS Well Plate will perform the next injection from the next well even if not all injections from that well in the programmed injection method have been executed User s Manual FAMOS D931R1 E 5 Digital Inputs and Outputs Freeze input The FAMOS Well Plate Microautosampler will freeze the analysis time for the time tha
60. be used to avoid loss of accuracy due to expansion of the loop content when switching from inject to load position prior to sample loading When working with high pressure 200 bar this loss may be up to 0 025 uL for a 5 uL loop IIe Note The minimum sample loss in partial loopfill mode is 5 uL recommended minimum flush volume for the first injection and an additional 2 5 uL always half the programmed flush volume for additional injections from the same well If a wash between injections has been programmed sample loss is 5 wL for every Injection For zero sample loss injections use the wL pick injection mode e pL Pick up means zero sample loss maximum accuracy Same as partial loopfill but slightly diminished reproducibility RSD better than 1 5 for injection volumes gt 1 uL Sample volume must be larger then 10 uL For optimum accuracy and reproducibility a 25 uL syringe and a 10 uL sample loop should be used The sample plug Is transported into the loop preceded by a plug of transport liquid that has a volume of 2 5 times the programmed needle tubing volume D931R1 User s Manual FAMOS APPENDIX B Programming Charts B 1 Overview of the Operating Program The details of the operating program for the FAMOS Well Plate Microautosampler is described in detail in Chapter 3 which includes a discussion of each parameter the allowable values and its role in the overall operation of the autosampler The table
61. bled 162000 fee te 2 4 uL for inert injection valve Inert Version PAEK Injection Valve Conventional Mode Inert conventional needle for FAMOS made of PEEK for inert 161474 cae ead injection valve User s Manual FAMOS D931R1 6 13 Maintenance and Troubleshooting 6 6 3 Syringe and Buffer Tubing Flangeless ferrule 1 16 blue for buffer tubing connection to low 160161 dispensing valve Male nut 1 16 blue for buffer tubing connection to low pressure dispensing valve 160160 Standard Version Stainless Steel Injection Valve Inert Version PAEK Injection Valve b includes Luer lock adapter P N 160137 syringe and buffer tubing 6 14 D931R1 User s Manual FAMOS Maintenance and Troubleshooting 6 6 4 Injection Valve Standard Version Stainless Steel Injection Valve Connecting tubing valve micro column 50 ul I D including low 160115 dispersion union Inert Version PAEK Injection Valve 161265 VALCO C2 6 Port PAEK Micro Injection Valve ASSY Replacement rotor for VALCO C2 6 Port Micro Injection Valve 161003 inert version 161004 Ta for VALCO C2 6 Port Micro Injection Valve inert 161263 162030 162031 161499 Connecting tubing valve micro column 50 ul I D including low dispersion union for inert injection valve 6 6 6 Vials and Accessories 160136 Polypropylene caps for FAMOS 1000 PCS User s Manual FAMOS D931R1 6 15 Maintenance and Troub
62. cape Key 3 4 lt EXCHANGE gt 3 10 lt EXIT gt 3 12 Executing a Program 3 6 Executing a Series 3 8 Remote Control 3 9 User s Manual FAMOS D931R1 Index F Factory Installed Items 2 3 Fan 2 5 Features of System 1 1 lt FIRST TRANSPORT VIAL gt 3 16 lt FLOW RATE gt 3 19 Fluid Connections 2 8 2 9 lt FLUSH VOLUME gt 3 18 lt FREEZE INPUT ACTIVE gt 3 16 Front Panel 3 3 lt FULL LOOP gt 3 18 Full Loop Injections 1 2 A 1 Function Keys 3 3 Fused silica needle 6 4 Fuses replacing 6 8 G lt GENERAL gt 3 13 General Design 1 3 General Menu 3 13 General Settings 3 14 H lt HEADSPACE PRESSURE gt 3 14 Help key 3 4 Hold Cont key 3 3 lt HOME gt 3 23 HPLC connections 2 8 I Initialization 3 2 lt INJECTION gt 3 18 Injection Principle 1 2 A 1 Injection Method 3 7 3 18 number 3 5 lt INJECTION CONTROL gt 3 19 lt INJECTION VOLUME gt 3 19 lt INJECT MARKER PULSE LENGTH gt 3 16 INPUTS 2 5 TTL E5 lt INPUT EDGE NEXT INJECTION gt 3 16 lt INSERT gt Mix Menu 3 21 User Program 3 22 Installation 2 1 Plate 4 3 Interrupt key 3 4 Introduction see System Overview 1 5 IO Menu 3 16 IO connections 2 5 lO Defaults 3 17 ISS Timebase 3 20 User Program 3 23 lt KEY CLICK gt 3 14 Keypad 3 3 Keys 3 3 L lt LABELED WELLS gt 3 15 lt LABELED WELL MARKER PULSE LENGTH gt 3 16 lt LAST TRANSPORT VIAL gt 3 16 lt L D FACTOR gt 3 19 Location in Laboratory 2 1 lt LOAD gt
63. ce use the long PEEK hex style nuts P N 161007 supplied with the instruments instead FIGURE G 4 u N FIGURE G 4 The Long PEEK Hex Style Nut i To use the supplied long hex nuts a Assemble the fitting as described in Section G 3 b Install the pre assembled fitting in the inert PAEK injection switching valve and manually turn the nut into the inert PAEK valve until it is finger tight FIGURE G 3 c Carefully tighten the hex style nut using a 14 wrench Caution Never use the inert PAEK injection switching valves to pre assemble a fitting and never use any tool to tighten the fingertight nut This may lead to a damage of the inert PAEK valve User s Manual FAMOS D931R1 G 3 Connecting the Nano Column G 5 Spare Parts Lists G 5 1 Tubing and Fittings PEEK Fingertight nuts and ferrules for inert injection switching valves 161000 set of 10 AE r 161007 Hex nuts and ferrules for inert injection switching valves set o G 4 D931R1 User s Manual FAMOS Index Ie Note Items indicated in Brackets and Capital Letters Refer to Commands on the Display lt uL PICK UP gt 3 18 HL pick up Injections 1 2 Air Segmentation A 8 Standard Mode A 6 A lt AIR SEGMENT gt 3 14 Air Segmentation uL Pick Up Injections A 6 Full Loop Injections A 3 Partial Loop Injections A 5 lt ALARM BUZZER gt 3 14 lt ANALYSIS TIME gt 3 18 Arrow Keys 3 3 lt ASPIRATE gt Mix Method 3 21 U
64. certain to indicate that a 10 pl loop is used in the system settings when wL pick up injections are selected Section 3 8 2 C 4 D931R1 User s Manual FAMOS Additional Programming Examples f Program the series by entering the parameters indicated in TABLE C 7 TABLE C 7 Programming Series Parameters Series to enter the Series Menu 01 E to define the series number 02 E to define the injection method for this series 01 E to define the wash method for this series lt ROW A gt 01 E to define the location of the first sample well lt ROW A gt 01 to define the location of the last sample well Escape to return to Ready Menu g Run the series by entering the parameters indicated in TABLE C 8 and pressing lt START gt TABLE C 8 Running the Series Start Stop to start the FAMOS Well Plate Microautosampler 01 E to start at series 01 01 E to stop after series 01 lt START gt to start execution of the series At the end of the defined analysis time the Ready Menu will appear again to indicate that the FAMOS Well Plate Microautosampler is ready for the following next run User s Manual FAMOS D931R1 C 5 Additional Programming Examples C 4 A 1 10 Dilution followed by a 1 0 uL Partial Loopfill Injection This example describes how to let the FAMOS Well Plate Microautosampler transfer 9 uL from Reagent A to the destination vial add 1 uL of sample followed by subsequently inject 1 0 uL 3 times
65. color at the AN back of the FAMOS Well Plate Microautosampler calls attention to the fact that you are expected to consult this manual for instructions on how to operate the FAMOS Well Plate Microautosampler X D931R1 User s Manual FAMOS CE DECLARATION OF CONFORMITY We LC Packings Nederland BV A Dionex Company Abberdaan 114 1046 AA Amsterdam The Netherlands declare that our product FAMOS Well Plate Microautosampler is in confirmation with the following documents EEC directives 89 392 incl 91 368 and 93 44 machine safety and EEC directives 73 23 and 93 68 low voltage safety applied with the following standard EN61010 1 Safety requirements for laboratory equipment Class I Installation cat Il Pollution degree II caused by connecting this instrument to devices which do not meet relevant AN LC Packings will not accept any liability for damages direct or indirect WARNING Safety standards EEC directives 89 336 and 92 31 EMC requirements applied with the following standards EN 55011 Radio frequency emission EN 50082 1 Voltage fluctuations EN 61000 3 2 Harmonic current emissions eae Use shielded cables and connectors for all remote connections Amsterdam January 11 2001 D932R1 Robert van Ling QA manager User s Manual FAMOS D931R1 Xi CE Declaration This page intentionally left blank xii D931R1 User s Manual FAMOS System Overview 1 1 Features and Design of
66. column The analysis time is started User s Manual FAMOS D931R1 A 7 Injection Principles lt Syringe 6E amp gt BZ Transport liquid Sample FIGURE A 15 The Injection Valve in the INJECT Position Aspirating the Programmed Injection Volume The next sequence will skip the first withdrawal of transport solvent unless a wash routine is performed or the FAMOS Well Plate Microautosampler has emptied the buffer tubing to waste In those cases the sequence is completely repeated A 3 2 Using an Air Segment If an air segment has been programmed it appears at the front of the first plug of transport liquid and at the front of every sample plug Use of an air segment can be enabled in the System Menu General Menu Note The air segment at the front of the sample plug is injected into the HPLC system Needle Injection loop Buffer fubing AA Transport Sample Air Fluent Transport Air gt B Transport Sample Eluent Transport ay A with air segment B without air segment FIGURE A 16 uL Pick up Injections with and without Air Segment A 8 D931R1 User s Manual FAMOS Injection Principles A 4 Low Dispersion Injection In addition to the standard injection methods the FAMOS Well Plate Microautosampler allows the analyst to control the injection profile via the Low Dispersion routine in the partial loopfill and the full loopfill
67. d ferrule nut item 1 FIGURE 6 2 c Carefully pull out sample needle and tubing d Remove the air outlet nut and replace it by a new one e Insert a new sample needle and tube through the needle holder and tighten the nut User s Manual FAMOS D931R1 6 5 Maintenance and Troubleshooting f Connect the other end of the tube to port 4 of the VICI Valco injection valve using a VICI Valco nut and ferrule item 1 FIGURE 6 2 Do not overtighten to prevent block of tubing g Lower the cover of the FAMOS Well Plate Microautosampler h Check sample needle height default height 2 mm from plate If necessary adjust the value in the System Menu General Menu refer to Chapter 3 i Select the lt WASH gt soft key in the Ready Menu to clean the new sample needle To install a different needle a Remove needle as described above b Remove the standard air outlet nut Item 6 FIGURE 2 3 item 6 and replace it by the nut supplied with the optional sample needle c Install the optional sample needle as described in Section 6 4 3 d Adjust settings in System Menu General Menu to the volume of the new needle tubing see section User nterface TABLE 6 3 Characteristics of Needles for FAMOS Well Plate Microautosampler mm mm mm mm mm mm 60117 b 0 375 0 15 585 tainless Steel Needles 10 3 uL 140 420 a inert version b extra long version Refer to Section 6 6 for an overview of
68. d use of a Mix Method in the System Menu Usage Menu you will also have to define the location of the First destination well and Reagent vials IES Note Series are stored in the microautosampler memory for as long as the power is on AS soon as power is switched off all programmed series will be deleted It is not possible to leave the Series Menu before all values have been programmed Calibration Sample OOVDOOOOODO OO OOO OOO OO OOOQ _ _ 0 OOM 020202020 OOO CD FIGURE 3 6 Injection sequence with 3 calibration wells between every 5 wells Users Manual FAMOS D931R1 3 27 The User Interface This page intentionally left blank 3 28 D931R1 User s Manual FAMOS CHAPTER 4 Programming Examples 4 1 Overview The commands described in Chapter 3 can be used by the analyst to generate a broad range of programs for the operation of the microautosampler These commands allow the user to configure the unit to meet the specific needs of the laboratory A series of blank forms are provided to assist the analyst in programming Appendix F This chapter describes the editing of general system settings and selection of the plate In addition a few simple programs that show the general approach to programming additional programs are presented in Appendix C are included While these programs may not meet the specific needs of the analyst it is likely that they can be used with minor modi
69. define location of the first sample vial lt ROW A gt 01 E to define location of the last sample vial Escape to return to the Ready M enu 4 4 D931R1 Users Manual FAMOS Programming Examples d Run the series by entering the parameters indicated in TABLE 4 4 and pressing lt START gt TABLE 4 4 Running the Series Start Stop to start the FAMOS Well Plate Microautosampler 01 E to start at series 01 01 E to stop after series 01 lt START gt to start execution of the series At the end of the defined analysis time the Ready Menu will appear again to indicate that the FAMOS Well Plate Microautosampler is ready for the next run 4 4 3 Example 2 Injecting a Single 100 nL sample from 3 vials and Injecting 5 uL samples from 3 other vials This example describes an application in which a single injection is made from each of three vials and a single injection with a different volume made from three additional vials a Place the appropriate vials in positions A1 A3 for the 100 nL Injections and the appropriate vials in positions C1 C3 for the 5 uL injections b Program the injection method for the 100 nL injections by entering the parameters indicated in TABLE 4 5 TABLE 4 5 Programming the Injection Method for 100 nL Injections Methods to enter the Methods M enu lt INJ ECTION gt 01 E to define 1 for the first method lt PARTIAL gt E to select partial loop fill 25 00 E to set analysis time to 25 m
70. e nut 1 4 turn 90 past the point where the pre assembled sleeve first start to grab the tubing FIGURE G 2 Use a 4 wrench to retain the union SF Note The amount of force to turn the nut can vary considerably due to the friction between the nut and the threads and as well as the composition and wall thickness of the tubing used Because of these variables we do not provide a torque specification G 2 D931R1 User s Manual FAMOS Connecting the Nano Column CAUTION CAUTION h Remove the fitting and inspect it The PEEK sleeve should be firmly attached to NANO column or the capillary respectively i e you should not be able to move it laterally along the tubing axis If the PEEK sleeve is loose reinstall the fitting in the union and tighten it another 1 8 of a turn past finger tight I Remove re inspect and repeat if necessary Flow rnrn TR ee PN 2 N aa A FIGURE G 3 Installing the Pre assembled Fitting j Install the pre assembled fitting in the inert PAEK injection switching valve and manually turn the nut into the valve until it is finger tight FIGURE G 3 Caution Never use the inert PAEK injection switching valves to pre assemble a fitting and never use any tool to tighten the fingertight nut This may lead to a damage of the inert PAEK valve G 4 Using Long PEEK Hex Style Nuts In case the supplied fingertight fittings cannot be used due to limited spa
71. e valve may be exceeded Check for possible leakage appears after every 50 000 syringe actions and after every 200 000 syringe valve actions lt LOG gt IES Note If you do not replace the Syringe when the message is presented and tell the system not to display this message again the message will not be displayed again until an additional 50 000 more syringe actions have been counted IES Note If the Synnge valve is replaced the counter should be reset to zero by the service engineer This key is used to change all software settings to default All lt DEAULTALL gt series methods templates and the user program unless protected by protection code will be erased TS Note If lt DEFAULT ALL gt is selected check whether the hardware configuration is still compatible with the settings entered in the System Menu Users Manual FAMOS D931R1 3 11 The User Interface lt SERIAL gt lt PANIC gt lt EXIT gt lt SSV gt option lt COOL gt option lt MANUAL gt lt AUTOMATIC gt lt DATE TIME gt lt SERVICE gt This key is used to set the FAMOS Well Plate Microautosampler to serial mode ae Note If a method protection code was defined in the system settings the code must be entered to get access to serial mode This key is used to initiate a stop sequence A panic stop does not remove the sample from the buffer tubing Before starting again you shou
72. e Method Methods to enter the Methods Menu 123456 E to enter the methods protection code lt TEMPLATE gt to enter the Template Menu 01 E to define the number for the template 02 E to define the injection method for this 01 template Escape Escape to define the wash method for this template to return to the Ready Menu b Program the series by entering the parameters indicated in TABLE C 15 TABLE C 15 Programming Series Parameters Series to enter the Series Menu 01 E to define the Series number 01 E to define the Template method number lt ROW A gt 01 E to define the first sample well lt ROW B gt 01 E to define the last sample well Escape to return to the Ready Menu c Run the series by entering the parameters indicated in TABLE C 16 and pressing lt START gt TABLE C 16 Running the Series Start Stop to start the Microautosampler 01 E to start analysis at series 01 01 E to stop after analysis of series 01 lt START gt to start the analytical run The FAMOS Well Plate Microautosampler now performs the same actions as in Example 2 except that analysis is performed on two wells A 01 and B 01 Re Note Select lt DEFAULT ALL gt in the Ready Menu Utilities Menu to erase all series and methods defined in these examples and to default all settings User s Manual FAMOS D931R1 C 9 Additional Programming Examples C 6 1ul injections out of 25 wells of a 384 well plate
73. e loop than other techniques 3 x loop volume for loop volumes lt 100 uL 2 x loop volume for loop volumes gt 100 uL 499 uL 1 5 x loop volume for loop volumes gt 500 uL INJ ECTION Ie Note Injection controls options are available in CONTROL Micro mode and only in conjunction with partial loopfill or full loop injection only Determines whether the injection is performed in the Is standard lt STD gt or in the low dispersion mode lt L D gt FLOW RATE Enter the flow rate L D FACTOR This entry is used to indicate the low dispersion factor range 0 7 2 0 ae Note These two parameters determine the Sw itching time of the injection valve refer to Appendix A for more details 3 8 11 The Method Menu Item lt WASH gt This soft function key is used to access a series of commands to program wash methods You can program a wash between injections samples or series When you select a wash method a screen will be presented that will allow you to indicate the volume of wash solvent can be defined The minimum programmable volume is 300 uL lt WASH gt Users Manual FAMOS D931R1 3 19 The User Interface 3 8 12 The Method Menu Item lt TIMEBASE gt This soft function key is used to enable control of the lt TIMEBASE gt optional ISS valve and other devices via auxiliary P5 or binary TTL outputs P3 if enabled in System Up to 5 timebase methods can be programmed The Menu Usage Menu m
74. ed to define the Solvent Selection option Valve SSV port position range 1 to 6 3 24 D931R1 User s Manual FAMOS The User Interface TABLE 3 3 Syringe Speed Speed asa oom 250m soom i000 a as sof as zo os 2em 31 3 s ais 630 ia 3 moman 625 250 625 1250 25 ra thigh 93 8 375 940 1080 3 8 rs asz5 mof 1920 ssf 77 rs 2675 1070 2675 sas 107 rs aes aras 2365 imine caution Caution The pressure in the buffer tubing will increase during the dispense action To prevent damage of the buffer tubing the flow should not exceed the value of 6mL min for water A maximum speed of 9 for 25 100 and 250 wL syringes a maximum speed of 6 for a 500 pL syringe and a maximum speed of 4 for 1000 uL syringe should be used If more viscous liquids are used the speeds should be reduced 3 8 15 The Method Menu Item lt TEMPLATE gt If the TEMPLATE option is enabled in the System menu the soft function key lt TEMPLATE gt is presented lt TEMPLATE gt Use this key to enter a menu in which the contents of a template can be defined First assign a number to the template then link the numbers of methods to the template User program The following items can be entered to fill a template instead of methods lt YES gt The complete template is filled with the user program no other methods can be added The template can be filled with the follow ing mix method number injecti
75. en An overview of the programming scheme is presented in Appendix B The different levels of the soft function keys commands on the different menus are indicated as follows lt PLATES gt lt bold with brackets gt represents the top level lt EXCHANGE gt lt normal type with brackets gt represents second level NEEDLE HEIGHT normal type represents the different parameter names 3 8 1 Ready Menu The Ready Menu contains the soft function keys presented in FIGURE 3 3 Overview lt PLATES gt lt WASH gt lt SYR END gt lt UTILS gt Second page of the Menu lt SERIAL gt lt SSV gt lt COOL gt lt SERVICE gt Only if option is installed FIGURE 3 3 The Ready Menu All available soft function keys and options on the Ready Menu lead to the following menu lt PLATES gt This key is used to access commands that are used to exchange well plates Press lt EXCHANGE gt to move the plate to the left in this lt EXCHANGE gt position the plate can be replaced without damage to the equipment lt PLATE HOME gt Press the soft function key lt PLATE HOME gt to move the plate to operating position again lt WASH gt This key is used to start a standard wash procedure All tubing connected to the syringe valve will be filled and rinsed with wash solvent This key is used to move the syringe to the end position e g if you wish to replace the syringe needle or to simplify filling of wash
76. enter lt NONE gt for wash method lt ROW A gt 01 E to define location A1 as first sample vial lt ROW A gt 01 E to define location A1 as last sample vial Escape to return to the Ready M enu g Run the series by entering the parameters indicated in TABLE 4 12 and pressing lt START gt TABLE 4 12 Running the Series Start Stop to start the FAMOS Well Plate Microautosampler 01 E to start at series 01 01 E to stop after series 01 lt START gt to start execution of the series At the end of the defined analysis time the Ready Menu will appear again to indicate that the FAMOS Well Plate Microautosampler is ready for the following next run 4 8 D931R1 Users Manual FAMOS diia Testing the Microautosampler 5 1 Overview This chapter describes a series of activities that can be used to check the operation of the system and to verify that your FAMOS Well Plate Microautosampler is operating in an acceptable manner This chapter includes e Establishing a Test Protocol Section 5 2 e Running the Test Protocol Section 5 3 User s Manual FAMOS D931R1 5 1 Testing The Microautosampler 5 2 Establishing a Test Protocol This section describes the parameters for the test protocol that will be performed in Section 5 3 A detailed discussion of how the parameters are accessed and edited is presented in Chapter 3 a Verify that the microautosampler is configured as listed in TABLE 5 1 and se
77. enu offers the following soft function keys This command scrolls through all program lines by pressing E or select AUX to move to the next auxiliary There are four AUX entries AUX1 AUX4 This command controls the ISS valve and the solvent selection valve The ISS valve can only be programmed if the optional ISS valve is installed The entries 6 1 and 2 1 refer to the interconnected ports of the valves Press E to scroll through programming lines Enter the time and the SSV port number value aw 1 and 6 aH Enter a time and a hexadecimal value betweenl and 15 Press E to scroll through the programming lines lt VALVES gt lt CODE gt This command is used to enter the end time for timed events program press E to scroll through the programming lines If no value is filled in or if CL is lt END TIME gt pressed the FAMOS Well Plate Microautosampler will automatically generate an end time The end time Is equal to the analysis time programmed in the injection method used in the same series eS Note If end time exceeds the programmed analysis time this end time overrules the analysis time You can program events after the end time but these events are not carried out during a run 3 20 D931R1 User s Manual FAMOS The User Interface 3 8 13 The Method Menu Item lt MIX gt If the MIX option is enabled in the System menu the soft function key lt MIX gt will be presented lt MIX gt if enab
78. essure on the sample approximately O 5 bar 6 4 D931R1 User s Manual FAMOS Maintenance and Troubleshooting Re Note Most commercially available sealers or capmats cannot be used in combination with headspace pressure We recommend that headspace pressure is switched off in those cases System Menu General Menu A schematic diagram of the needle assembly is shown in FIGURE 6 2 and a list of the optional needles is presented in TABLE 6 2 and TABLE 6 3 1 Nut and Ferrule 2 Needle Tubing 3 Needle Connection Nut 4 Conventional Sample Needle Z 5 Needle Holder cas eel L 6 Air Outlet Nut a i 7 P turing Need repuncturin eedle O T eee puncturing 8 Sensor L constitute the sample needle FIGURE 6 2 Needle Assembly TABLE 6 2 Optional Needles Needle Type Fused silica needle needle with small inner diameter for small injection volumes samples or in case large volumes are loaded in the loop the FAMOS Well Plate Microautosampler Refer to Section 6 6 for an overview of options available for the FAMOS Well Plate Microautosampler To replace the present needle a Loosen the needle connection nut item 3 FIGURE 6 2 b Loosen the VICI Valco an
79. eturn to the Ready Menu Ie Note Reagent and transport vials can be placed in any of the four positions Transport vials must be placed in a continuous row User s Manual FAMOS D931R1 2 13 Installation and Getting Started This page intentionally left blank 2 14 D931R1 User s Manual FAMOS CHAPTER 3 The User Interface 3 1 Overview This chapter describes the general mode of operation of the FAMOS Well Plate Microautosampler and explains how an operating program is established It includes a discussion on e Powering Up the Microautosampler Section 3 2 e A description of the front panel and the role of the keys on the front panel Section 3 3 e Menus of the FAMOS Well Plate Microautosampler Section 3 4 e General approach to entering and executing a program Section 3 5 e Types of Methods Section 3 6 e Executing a Series Section 3 7 e Menus and Operating Commands Section 3 8 e Series Menus Section 0 Typical programs are presented in Chapter 4 and Appendix C Users Manual FAMOS D931R1 3 1 The User Interface 3 2 Powering up the FAMOS Well Plate Microautosampler a When the FAMOS Well Plate Microautosampler is powered up via the main power switch on the rear panel it will go through an initialization self test protocol During this period a number of messages are displayed indicating that various components are functioning properly b After completion of this procedure t
80. f you have programmed a method and defined a series for the samples you wish to analyze Note Series are not stored in battery backup memory and exist only for as long as the FAMOS Well Plate Microautosampler is powered up To execute a series a Start programming the series by pressing Start Stop b Enter the number of the first series to be performed and press E to confirm the input and go to the next screen then indicate the number of the last series to be performed and press E again c After the range of series is defined the autosampler will ask for the start conditions It is possible to start the series either from the keypad or by remote control inputs NEXT INJ ECTION and NEXT VIAL inputs connector P6 d Select lt START gt to manually start the analytical run The FAMOS Well Plate Microautosampler will begin execution of the series that you have defined During the run the display shows information about the current operation Note Series are always executed in numerical order Empty series will be skipped You can stop a run by pressing the Start Stop key The FAMOS Well Plate M icroautosampler will execute a shut down sequence by removing all sample from the buffer tubing and performing a wash routine If you want to stop the microautosampler immediately panic stop without performing a buffer clean up press the Start Stop key twice Remember to perform a wash routine to clean up the tubing before you sta
81. fication e g parameters such as the analysis time and sample volume may need to be changed An additional rationale for the presentation of these programs Is to provide a description of the logical processes that are used in generating a program It should be noted that the examples described below could be used with systems that include UltiM ate with the UltiChrom software program as well as for stand alone systems The following programs are included in this chapter e Single 200 nL injection Shows partial loopfill Section 4 4 2 e 100 nL injection from of 3 vials and 5 nL injections from 3 other vials shows partial loopfill and injections from multiple vials Section4 4 3 e 1 uL injection out of a 5 uL sample in vial 1 using transport liquid Shows uL pickup Section 4 4 4 Additional examples are provided in Appendix C Users Manual FAMOS D931R1 4 1 Programming Examples 4 2 General System Settings When the microautosampler is initially installed a variety of general parameters are set which generally define the configuration of the unit and should be checked When you are defining a new method you should check these values to make sure that they are relevant To check the general system settings press the System key then select the soft function keys lt MICRO gt and lt GENERAL gt as appropriate and press the E key on a sequential basis to view each selection Use the E key to go through the
82. general system settings A set of reasonable values is presented in TABLE 4 1 Note The E key is used to indicate that the user accepts the indicated value for a parameter and he she wants to access the next parameter field For the sake of brevity it will be understood that the E key must be pressed when a parameter is view ed edited TABLE 4 1 Typical General Parameter Settings lt GENERAL gt Parameter Volume of the installed injection loop Volume of the tubing needle valve 2 4 uL Syringe speed Scale factor O To go back to the Ready menu press Escape twice Note Adjust the needle height to the desired depth of insertion Use 4mm 04 for the enclosed 250 uL polypropylene tapered vials 4 2 D931R1 Users Manual FAMOS Programming Examples 4 3 Installing the Plate To install the plate a Insert the 48 vial holder FIGURE 4 1 It will be necessary to use the lt EXCHANGE gt soft key to move the plate holder to the left to access the plate holder FIGURE 4 1 48 Position Well Plate b Press the lt PLATE HOME gt soft key to move the plate holder back into Operation position and place a sample vial in position Al To check the plates setting a Press the System key and then select the soft function keys lt MICRO gt and lt PLATES gt b Select lt 48 vials gt c Select Process Vials lt IN ROWS gt To go back to the Ready Menu press Escape twice Users Manual
83. gt lt USRPROG gt Methods Userprog or lt INJECTION gt lt WASH gt lt TIMEBASE gt lt MIX gt ASPIRATE Mix method DISPENSE Injection method SYR VALVE Waheed Tajeation methods SYRINGE wee ees e Wash volume 4x AUX 1 e ASPIRATE me e Wash between 4x AUX 2 sample WAIT Series 4x AUX 3 destination _ Wells 4x AUX 4 air COMPRESS lt FLUSHED gt lt PARTIAL gt lt PICK UP gt sigs AUX Injections 4x ISS A reagent L L 4x ISS B DISPENSE WAIT INPUT Analysis time Analysis time 3x CODE OUT sample PROG OUT Flush volume Flush volume END TIME destination CODE Injections well Injections well waste MARKERS Inject control Inj volumes reagent STDIL D Inject control REPEAT STD L D WAIT WASH Flowrate Flowrate Delay factor Delay factor With templates Series Template Userprogram yes no Use calibration wells yes no or calibration wells Mix method calibration interval Injection method Sample wells Wash method Destination wells Time base method Reagent wells Use calibration wells yes no When the conventional autosampler set up is selected in the Labeled wells calibration wells system setting the value ranges of the system settings and calibration interval in the methods will be adjusted to the needs of the Sample wells conventional autosampler Destination wells Also will the Low Dispersion mode in the injection methods Reagent wells no longer be available Labeled wells Note
84. gt for wash method lt ROW A gt 01 E to define location of the first sample well lt ROW A gt 01 E to define location of the last sample well Escape to return to the Ready Menu e Run the series by entering the parameters indicated in TABLE C 4 and pressing lt START gt TABLE C 4 Running the Series Start Stop to start the FAMOS Well Plate Microautosampler 01 E to start at series number 1 01 E to stop after execution of series number 1 lt START gt to start the analytical run The FAMOS Well Plate Microautosampler will now locate well AO1 and perform a 1 0 uL partial loopfill injection The display of the FAMOS Well Plate Microautosampler will indicate the status Checking tray Flushing Loopfill Running Rinse buffer Running The display also indicates the number of the defined series 01 the method number 01 and the well on which the analysis is performed A01 At the end of the defined analysis time the Ready Menu will be displayed again to indicate that the microautosampler is ready for the next analytical run User s Manual FAMOS D931R1 C 3 Additional Programming Examples C 3 A3x 1 0 uL Injection with uL Pick Up and Wash between Injections This example describes an application where three 1 0 uL injections are made with a wash between each injection a Place a vial with transport solvent mobile phase in transport vial position 1 left and place the sample in position A1 Make sure t
85. h volume FIGURE A 9 This air segment is at the front of the flush volume and will not be injected Use of an air segment can be enabled in the System Menu General Menu Needle Injection loop Buffer tubing A Sample Elueru flush Air A E B Sample Eluent Flush gt LIL LMM Ee A with air segment B without air segment FIGURE A 9 Partial Loop Injection with and without Air Segment User s Manual FAMOS D931R1 A 5 Injection Principles A 3 uL Pick Up Injections A 3 1 Standard Mode The switching sequence for a uL pick up injection is schematically presented in this section At the start of the sequence the injection valve is in INJECT position FIGURE A 10 The sample needle has entered the vial of transport liquid normally mobile phase to avoid disturbance of the chromatogram with an additional peak of the transport solvent after the air needle has pre punctured the septum The headspace pressure applied through the outer air needle ensures that no air or vapor bubbles are formed during wash solvent aspiration amp Syringe gt Transport liquid FIGURE A 10 Injection Valve in Inject Position Sample Needle in the Transport Vial Re Note When performing uL pick up injections use the mobile phase as the transport liquid to avoid a disturbance of the chromatogram due to an additional peak of the transport solvent For the first injection after a wash or after emptying of
86. hat is installed and adjust if necessary System Menu General Menu To replace a plate in the FAMOS Well Plate Microautosampler a Select the lt PLATES gt soft key in the Ready Menu then select lt EXCHANGE gt The plate moves to the left b Take out the plate and replace it by another one c Select the lt PLATE HOME gt soft key The plate moves to operating position again d If you have replaced the plate by a plate of the same type you are now ready If you have installed a new type of plate execute the following steps Press System Select the lt PLATES gt soft key Press E Select the soft function key for the type of plate concerned Press E and determine whether to process the plate in lt ROWS gt or in lt COLUMNS gt Press Escape twice to return to the Ready Menu A message appears to indicate that all programmed series will be reset The user will have to redefine series because the settings in the System Menu have been altered 2 6 3 Placing Reagent Vials Transport Vials in Position To replace reagent vials transport vials a Select the lt PLATES gt soft key and then select lt EXCHANGE gt in the Ready Menu The plate holder moves to the left b Take out the reagent vials transport vials item 7 FIGURE 2 1 and replace them by other reagent vials transport vials c Select the lt PLATE HOME gt soft key The plate moves to operating position again e Press Escape twice to r
87. he FAMOS Well Plate Microautosampler is a Enter the desired settings in the System Menu It is probable that the settings in the System menu have been correctly entered factory installed as these settings are global in nature b Program a method for the analyses you wish to perform via the Methods Menu c Define a series and link a programmed method to a range of wells in the Series Menu d Execute the series While you could use a different order than that indicated above it should be noted that some of the settings in the System Menu define the options in other menus If an invalid selection is made on the System Menu the desired option for other menus may not be presented As an example of this point the USER PROGRAM menu which is defined in Section 3 8 14 will be presented only if the USER PROGRAM option has been selected in the System Menu Section 3 8 2 3 6 D931R1 User s Manual FAMOS The User Interface 3 6 Types of Methods The FAMOS Well Plate Microautosampler provides the following types of methods for different aspects of the operation of the microautosampler e injection method contains information on the injection routine flush volume and analysis time e wash method describes a wash volume and when a wash must be executed e mix method a pre injection method in which additional sample handling can be performed e g pre column derivatization e timebase method a post injection method w
88. he Ready Menu appears on the display 14 46 THURSDAY JANUARY 04 01 MENU READY 2 02 lt PLATES gt lt WASH gt lt SYREND gt lt UTILS gt FIGURE 3 1 The Ready Menu eS Note In this manual the various display messages and menus shown correspond to a system with firmware version V2 02 If a different version of the firmware is used there may be small differences in the screens and or actions that occur when a given command is performed 3 2 D931R1 User s Manual FAMOS The User Interface 3 3 The Upper Front Panel Communication between the user and the system Is provided by the keypad on the front panel of the FAMOS Well Plate Microautosampler FIGURE 3 2 or via the UltiChrom Software This section describes the display and the use of the keypad when the microautosampler is used on a local basis i e not controlled by UltiChrom Software 13 38 TUESDAY HOUEMBER 14 69 CMEHUT READY 2 823 Display lt PLATES gt lt WASH gt SYR ENDO lt UTILS3 Soft Keys Arrow Keys Numeric Keys Function Keys FIGURE 3 2 Keypad and Display The front panel includes e Display indicates a variety of system parameters e Soft Keys the label assigned to these keys depends on the menu that is active The current function of each key is shown in the bottom line of the display e Arrow keys used to move to a different field in the display to move to a different field in a menu or to change the value of
89. he scale factor multiplied by the syringe speed The speed of the syringe during the wash or the rinsing procedure of the buffer are not affected by this setting SYRINGE SPEED SCALE FACTOR Users Manual FAMOS D931R1 3 13 The User Interface This parameter refers to the distance between the needle point and the plate holder default 2 mm The value ts only used in injection methods for mix methods this value is programmable in the method itself NEEDLE HEIGHT Appears only if a plate with 48 vials is selected in System Menu Plates Menu YES means that empty SKIP MISSING VIALS Spaces are skipped during the run NO means that the Microautosampler will stop if an empty space is observed during the run and an error code will be generated This command is used to indicate whether an air AIR SEGMENT segment will be used for analytical runs for explanation of air segmentation refer to Appendix A This command is used to indicate if the headspace pressure should be on or off The FAMOS Well Plate Microautosampler uses headspace pressure to facilitate transport of sample into the loop The compressor will HEADSPACE always be used during a wash procedure PRESSURE eS Note Accuracy and reproducibility may decrease if headspace pressure is switched off However headspace pressure will only be useful if sample wells are airtight This command is used to offer a choice between two EAS EE modes of representing the time KEY
90. he transport vial is correctly filled before starting a new series b Program the System Parameters by entering the parameters indicated in TABLE C 5 TABLE C 5 Programming System Parameters System to enter the System Menu lt MICRO gt E to select the micro mode lt GENERAL gt E to enter the General Menu E until Air segment to go to the Air segment field appears lt NO gt E to switch off air segment Escape to return to the System Menu lt PLATES gt E to enter the Plates Menu EE to go to transport vials field 01 E to define position of the first transport vial 01 to define position of the last transport vial Escape Escape to return to the Ready Menu e Program the method by entering the parameters indicated in TABLE C 6 TABLE C 6 Programming Method Parameters Methods to enter the Methods Menu lt INJECTION gt 02 E to define method number 02 lt PICK UP gt E to select the injection mode for this method 100 E to define the analysis time 3 E to define the number of injections per well 100 E to define volume of 1 0 uL for 1 injection 100 E to define volume of 1 0 uL for 2 injection 100 E to define volume of 1 0 uL for 3 injection Escape to return to the Methods Menu lt WASH gt to enter the Wash Menu 01 E to define wash method number 01 lt INJECTION gt E to select wash between injections 50 to define the wash volume Escape Escape to return to the Ready Menu IES Note Make
91. hed using the system flush it with a water methanol or water acetonitrile mobile phase before shutting it down e Solvent should be degassed by sparging with He 2 6 2 Plates and Sample Handling The FAMOS Well Plate Microautosampler accommodates the following types or plates e 96 low wells e 96 high wells e 384 low wells e 48 vials A sensor item 5a FIGURE 2 1 monitors plate detection plate height detection and vial detection Because the FAMOS Well Plate Microautosampler uses headspace pressure during sample injections it is very important that samples are properly handled Note the following e Standard wells can best be filled by means of a narrow end pipette to allow air to escape when filling the well e If wells are filled to the rim the sample might be forced into the prepuncturing needle causing cross contamination of samples and contamination of the sample needle e If vials are used with a plate for vials make sure the seals are airtight to prevent air bubbles in the sample and prevent evaporation of volatile samples check seals after crimping if the cap can be turned easily the seal is not airtight and the handcrimper should be adjusted Note If wells that are not airtight are used switch off the headspace pressure in the System Menu General Menu D931R1 User s Manual FAMOS Installation and Getting Started Ie Note Check whether the needle height is sufficient for the new type of plate t
92. in 5 0 E to set the flush volume to 5uL 01 E to set the number of injections vial to 1 01 E to set injection volume to 0 1 uL lt STD gt E to select the standard injection control Escape to return to the Methods M enu c Program the injection method for the 5 uL injections by entering the parameters indicated in TABLE 4 6 TABLE 4 6 Programming the Injection Method for 5 uL Injections lt INJ ECTION gt 02 E to define 2 for the second method lt FLUSHED gt E to select flushed loop fill 25 00 E to set analysis time to 25 min 5 0 E to set the flush volume to 5uL 01 E to set the number of injections vial to 1 lt STD gt E to select the standard injection control Escape Escape to return to the Ready M enu Users Manual FAMOS D931R1 4 5 Programming Examples d Program the series by entering the parameters indicated in TABLE 4 7 TABLE 4 7 Programming the Series Series to enter the Series Menu 01 E to define the Series number 01 E to select the injection method number CLE to enter lt NONE gt for wash method lt ROW A gt 01 E to define location A1 as first sample vial lt ROW A gt 03 E to define location A3 as last sample vial Series to enter the Series M enu 02 E to define the next Series number 02 E to select the next injection method number CLE to enter lt NONE gt for wash method lt ROW C gt 01 E to define location C1 as first sample vial lt ROW C gt 03 E to define
93. in generating a program The following programs are included in this appendix e A 1 0 uL partial loopfill Section C 2 e A3x 1 0 uL injection with uL pick up partial loopfill injection Section C 3 e A 1 10 dilution followed by a 1 0 uL partial loopfill injection Section C 4 e Defining a Template and adding a protection code Section C 5 e 1 uL injections out of 25 wells using a 384 well plate Section C 6 e Column Switching Section C 7 It should be noted that the examples described below could be used with systems that include U tiMate with the U tiChrom software program as well as for stand alone systems User s Manual FAMOS D931R1 C 1 Additional Programming Examples C 2 A 1 0 uL Partial Loopfill This example describes an application in which a 1 0 uL injection is made from a single vial in position 1 In this example we assume that a 5 uL loop a 2 4 um needle and a 25 uL syringe have been installed a Place a sample in position A1 b Program the System Parameters by entering the parameters indicated in TABLE C 1 TABLE C 1 Programming System Parameters System to enter the System Menu lt MICRO gt E to select the micro mode lt GENERAL gt E to enter the General Menu 05 0 E to define the volume of the installed loop 02 4 E define the volume of the needle tubing lt LOW gt E set syringe speed to low 02 E set sample needle height to 2 mm lt NO gt E enable use of air segment lt YES gt E s
94. in the System Menu and the correct buffer tubing is installed refer to Section 2 3 An incorrect setting may lead to damage to the system g Select the lt WASH gt soft key in the Ready Menu to execute a standard wash routine All tubing connected to the syringe valve is filled and rinsed 6 3 2 Replacing the Syringe Tip To replace the tip of the syringe a Remove the syringe as described in Section 6 3 1 items a c b Remove the syringe tip c Place a new tip on a flat surface e g workbench making certain that attachment point is facing up d Gently push the syringe shaft in the new tip CAUTION Caution If you are installing a 25 uL syringe tip be extremely careful as the shaft is very thin and can be easily bent e Carefully insert the shaft in the syringe body f Re install and flush the syringe as described in Section 6 3 1 item d to g g Check for any leakage 6 3 3 Needle Assembly The sampling needle consists of two parts e Sample needle placed inside the hollow prepuncturing needle used for the actual transport of sample Different types of needles can be used see Section 6 6 2 If a needle with different diameter is used a different air outlet nut item 6 FIGURE 6 2 must be used that matches the injection needle The fused silica needle P N 160116 is the default needle e Prepuncturing needle a hollow needle used for puncturing of the septum Capmat or sealer also used to put headspace pr
95. ing Time sec f L D Factor Vi Injection Volume uL F Flow Rate wL min As an example If Vin 1 uL the flow rate is 5 uL and the L D factor is 1 1 the valve switching time S will be 13 2 s Note Under these conditions the loop will be flushed with the wash solvent as well which may result in injection problems e g lack of proper focussing It is recommended that the mobile phase typically mobile phase A is used as the wash solvent Note The injection time must be at least 10 s If the calculated injection time is less than this limit it will be redefined to 10 s User s Manual FAMOS D931R1 A 9 Injection Principles A 5 Comparing the Various Sample Injection Modes The characteristics of each injection mode is described below e Full loop injection provides maximum reproducibility RSD lt 0 4 but not maximum accuracy since loop volume is specified with an accuracy of 10 Minimum sample loss 12 4 uL 2 x loop overfill flush volume for needle for the standard 5 uL loop e Partial loopfill provides maximum accuracy depends on syringe accuracy and reproducibility better than 0 6 RSD for injection volumes gt 1 WL Minimum sample loss Flush volume 5 0 uL 5 0 uL is the recommended minimum flush volume smaller flush volumes can be programmed but will result in decreased performance For maximum reproducibility and accuracy a 25 uL syringe should be used and a 1 uL sample loop should
96. ing your unit Please feel free to photocopy the forms in this section User s Manual FAMOS D931R1 F 1 Programming Forms System Menu settings lt GENERAL gt lt USAGE gt Loop volume uL Protection code Needle tubing 1 Timebase volume H methods Syringe volume lenabled disabled uL Mix methods lenabled disabled Syringe speed User program Jenabled disabled factor User program sete protection code oe see wes _lyes Labeled wells lenabled disabled Air segment lyes Templates _lenabled _ disabled Headspace Calibration rere _lyes foal lenabled disabled Time display HH MM SS J H MM mm Key click lon loff Error beep lon loff Alarm buzzer lon loff lt PLATES gt 96 low Inject marker 96 high pulse length 1384 low Well marker 48 vials pulse length Labeled well processing In ROWS marker pulse length Input edge next ae _ COLUMNS niecaen _ falling _ rising position first Input edge next i transport vial well falling rising position last Freeze input Few high transport vial active J Reset outputs es no after last series Lly lt CLOCK gt lt COMM gt Off _ On yy mm dd and hh mm Device identifier Dati Comments Needle height F 2 D931R1 User s Manual FAMOS Programming Forms Templates Template Injection Wash Timebase User program Comments number method method method
97. inge cat no FMS SY 1000 a 2000 uL buffer tubing cat no FMS BT 2000 a 250 uL injection loop a standard injection needle assembly cat no FMS NDL CNYV installed e An additional HPLC pump for loading the sample at a flow rate of 0 5 mL min e A u precolumn holder cat no HD O5 and precolumns e g 500 um I D x 5 mm 5 um C18 cat no MCA 50 C18 To prepare the system e Replace the injection needle assembly Section 6 3 3 e Replace the 25 uL syringe for the 1000 uL syringe Section 6 3 1 and replace the buffer tubing e Connect the contact closure input of the data system to the AUX 1 contact of the autosampler P5 connector in order to start the data acquisition at the moment the pre column is switched on line e Set the system up according to FIGURE C 1 sample loop pump 4 pl min pump 0 5 ml min Capillary Column y precolumn 5mm x05 mm l D fused silica injection sample vial syringe FIGURE C 1 Column Switching Setup C 12 D931R1 User s Manual FAMOS Additional Programming Examples e Set the FAMOS Well Plate Microautosampler in lt CONVENTIONAL gt mode and adjust the general system parameters according to the modified setup e g syringe loop volume Section 4 2 e Enable the use of time based methods to allow programming of the AUX 1 contact closure Section 3 8 4 and 3 8 12 e Program injection method 01 with the following parameter
98. inge is available To replace the syringe a Press the lt SYR END gt soft function key in the Ready Menu to move the syringe to the end position and then lift the cover b Unscrew the top of the syringe turn clockwise Pull the syringe down until it releases from the syringe valve Then pull the syringe towards you and remove it FIGURE 6 1 c Unscrew the Luer Lock adapter 100 250 500 or 1000 uL syringe only 1 Syringe Valve offo N Luer Lock Adapter dae Syringe Syringe Tip oH Y Syringe Shaft lies 100 250 500 1000 uL syringes only FIGURE 6 1 Replacing the syringe d Attach the Luer Lock adapter to the new syringe 100 250 500 or 1000 uL syringe only IIe Note The 100 250 500 and 1000 uL syringes require the Luer Lock adapter P N 160137 User s Manual FAMOS D931R1 6 3 Maintenance and Troubleshooting CAUTION e Fill the new syringe with wash solvent and make sure that all air bubbles are removed from the syringe Connect the bottom of the filled syringe to the FAMOS Well Plate Microautosampler Screw the top of the filled syringe to microautosampler counter clockwise f Lower the cover Press the lt SYR HOME gt soft function key to remove air from the syringe The syringe moves to the home position and the content is dispensed to waste Caution Before continuing check if the correct syringe is selected
99. injection modes The injection principle is used to improve the injection profile and therefore improve the reproducibility by switching the injection valve back into load position after a calculated time and to cut off the tailing part of the sample plug Two parameters are used to control the Low Dispersion mode Flow Rate This value determines the flow rate at which the sample is flushed out of the injection loop This value corresponds to the flow rate of the LC system The valid range is 00 00 to 99 99 uL min Note If the flow rate is set to 00 00 uL min a standard injection is performed Low Dispersion Factor L D factor This value determines the amount of sample volume which is cut off by the back switching injection valve A value of 1 0 corresponds to the volume of the sample plug If the value is greater than less than 1 0 the injection valve will switch back after the volume is greater than less than the sample volume which has been transferred through the sample loop The valid range is 0 7 to 2 0 As an example if the value is 1 2 the valve will switch back after 120 of the value of the sample plug has been transferred The injection profile for various values is shown in FIGURE A 1 7 13 1 11 0 09 L D factor FIGURE A 17 Low Dispersion Injection Profiles The time the injection valve is switched back into the load position is calculated by the equation A 1 St f V1 60 F A 1 where St Switch
100. ion Section 6 3 Once you have remedied the problem repeat the performance test 5 4 D931R1 User s Manual FAMOS Testing The Microautosampler 5 4 Checking System Components 5 4 1 Testing the Accuracy of Delivery of the Syringe To determine the accuracy of delivery of the syringe dispense a volume of 25 uL of water from a sample vial to a destination vial program the following Mix Method TABLE 5 5 and Series TABLE 5 6 TABLE 5 5 Mix Method 1 Step Action Speed Height Dispense 25 uL to Destination End of mix method TABLE 5 6 Series 1 ROW A 01 To determine the accuracy of delivery of the syringe a Weigh the destination vial before and after the Run b The difference is the aspirated volume The specified variation is 2 5 4 2 Testing the Loop Volume To determine the volume of the injection loop a Disconnect the loop from the injection valve b Remove all liquids from the loop with air c Weight the empty loop on an analytical balance d Fill the loop with minimal 2 times its volume of water e Weight the filled loop again The difference in the weight of the loop divided by the specific weight of water 1 g mL gives the calibrated volume of the loop The specified variation is 10 User s Manual FAMOS D931R1 5 5 Testing The Microautosampler 5 4 3 Testing the Tray Cooling Option To test the tray cooling option if installed a Remove the plate holder
101. is interfaced to equipment from other manufacturers the user should refer to Appendix E for a detailed discussion of the various inputs and outputs that are provided 2 4 2 P4 Connector MARKERS If the microautosampler is installed with an U tiMate system the P4 connector should be connected to the START IN input of the UV Detector on the rear panel of the U tiMate system To connect the P4 MARKERS outputs use the Inject Marker Cable 2 4 3 P5 Connector AUXILIARIES If the microautosampler is installed with a Switchos II Advanced Micro Column Switching Unit and the U tiMate system the position of micro valves of the Switchos Il as well as the gradient start are controlled via the P5 connector To connect the P5 AUXILIARIES outputs to the Switchos II and the U tiMate system use the special interface cable P N 1601 71 2 4 4 P1 P2 P3 and P6 Connectors This connectors are not used during an standard installation with the U tiMate system refer to Appendix E for more details when the microautosampler is used with other systems 2 4 5 Communication Connector The FAMOS is equipped with two 9 pin RS 232 serial interfaces In the standard setup only the male type connector S2 is used for digital transfer between the autosampler and the PC item 1 FIGURE 2 2 Some communication parameters can be changed via two DIP switches above the RS 232 connectors These switches should be set to the default settings presented in FIGURE 2
102. is key can only be used if MENU or MN is shown in the top right hand corner of the display If this key is pressed additional fields of the menu are displayed e System used to enter the System Menu in which system settings can be entered c General keys e Escape allows the user to leave the programming mode or go to a previous level in the menu A value that is entered is checked for validity and then saved e Help used to display help information which is available only for a limited number of functions 3 4 D931R1 User s Manual FAMOS The User Interface 3 4 Menus of the FAMOS Well Plate Microautosampler The software of the FAMOS Well Plate Microautosampler is menu driven In this section we present an overview of the menu structure as well as a detailed description and a menu reference name for each command Section 3 8 In addition a detailed programming flow chart is presented in Appendix B To access a menu click on the soft key that corresponds to the menu A few menus have a sub menu associated with it in these cases the desired menu is chosen in the standard way after the top level menu is accessed When you are in a menu various items are accessed and you should indicate the appropriate choice s to generate the desired method or series The microautosampler menus are e Ready Menu this menu appears when the FAMOS Well Plate Microautosampler has been powered up and passed the initialization
103. ith which outputs to other devices e g integrator or pump and switching of the ISS valve are controlled e user program offers the possibility to program sequences of all actions that can be executed by the FAMOS Well Plate Microautosampler in separate steps Each programmed method Is assigned a number The FAMOS Well Plate Microautosampler allows you to store a combination of defined methods ina template which is also identified by a number Methods must be linked to a series before they can be used The follow ing Options are available with the FAMOS Well Plate Microautosampler e You Can assign an individual method to a series methods mix injection wash timebase can be linked to wells in a series e You can assign a template to a series a combination of various programmed methods mix injection wash timebase can be defined in a template The template is linked to a range of wells in a Series In this way all steps in an analytical run are defined and stored e You can assign a user program to a Series This can be used to combine all possible steps in the analytical process in one program The user determines the order of the separate actions the FAMOS Well Plate Microautosampler has to perform Users Manual FAMOS D931R1 3 7 The User Interface 3 7 Executing a Series 3 7 1 Manually Executing a Series A series can be run from the Ready Menu by pressing the Start Stop key Execution of a series is possible i
104. jection method number to enter lt NONE gt for wash method to define location G14 as first sample vial to define location G22 as last sample vial to enter the Series Menu to define the next Series number to select the next injection method number to enter lt NONE gt for wash method to define location M3 as first sample vial to define location M8 as last sample vial to return to the Ready Menu D931R1 User s Manual FAMOS Additional Programming Examples e Run the series by entering the parameters indicated in TABLE C 19 and pressing lt START gt TABLE C 19 Running the Series Start Stop to start the FAMOS Well Plate Microautosampler 01 E to start at series 01 03 E to stop after series 01 lt START gt to start execution of the series At the end of the defined analysis time the Ready Menu will reappear to indicate that the FAMOS Well Plate Microautosampler is ready User s Manual FAMOS D931R1 C 11 Additional Programming Examples C 7 Column Switching The FAMOS Well Plate Microautosampler is also available with an additional ISS valve Integrated Stream Switch option which can be used for column switching experiments In this section we provide an example of the use of a pre column prior to the capillary HPLC analysis The following equipment is needed to perform this column switching example e A FAMOS Well Plate Microautosampler with the ISS A valve ISS in position A a 1000 uL syr
105. l item 5 FIGURE 2 2 CAUTION Caution Make certain that the instrument is properly grounded to a true earth ground Connecting the instrument to an ungrounded power line can cause injuries and or damage the instrument User s Manual FAMOS D931R1 2 7 Installation and Getting Started 2 5 Connecting the Autosampler to the HPLC System 2 5 1 Preliminary Operations To install the FAMOS Well Plate Microautosampler to an HPLC system e Place the FAMOS Well Plate Microautosampler in its operating location preferably on the left hand side of the HPLC system Make sure the ventilation holes are not obstructed Allow the instrument to acclimatize for 1 hour e Loosen the screw at the right hand side of the cover item 6 FIGURE 2 1 and lift the cover so that you can perform the procedures described in this chapter e Install the Plate Holder item 1 FIGURE 2 5 in the FAMOS Well Plate Microautosampler It should be placed on the cooling plate item 3 FIGURE 2 5 underneath the wash solvent bottle as far to the left and to the back as possible as shown in FIGURE 2 5 The gear wheel item 2 FIGURE 2 6 must fit against the teeth of the plate holder indicated in white in the center of FIGURE 2 6 FIGURE 2 5 Installing the Plate Holder FIGURE 2 6 Bottom View of Plate Holder e Connect the HPLC pump to port 1 of the VICI Valco injection valve e Connect HPLC column to port 6 of the VICI Valco injection valve 2 8
106. l Plate Microautosampler 3 3 The Upper Front Panel 3 4 Menus of the FAMOS Well Plate Microautosampler 3 5 General Approach to Entering and Executing a Program 3 6 Types of Methods 3 7 Executing a Series vii IX XI 3 7 User s Manual FAMOS D931R1 Table of Contents CHAPTER 4 CHAPTER 5 CHAPTER 6 3 8 3 9 3 7 1 Manually Executing a Series 3 7 2 Executing a Series via Remote Control 3 8 1 3 8 2 3 8 3 3 8 4 3 8 5 3 8 6 3 8 7 3 8 8 3 8 9 3 8 10 3 8 11 3 8 12 3 8 13 3 8 14 3 8 15 Ready Menu The System Menu The System Menu Item The System Menu Item The System Menu Item The System Menu Item The System Menu Item The System Menu Item Methods Menu Menus and Operating Commands lt GENERAL gt lt USAGE gt lt PLATES gt lt O gt lt CLOCK gt lt COMM gt The Method Menu Item The Method Menu Item The Method Menu Item The Method Menu Item The Method Menu Item The Method Menu Item lt INJECTION gt lt WASH gt lt TIMEBASE gt lt MIX gt lt USER PROGRAM gt lt TEMPLATE gt Series Menu Programming Examples 4 1 4 2 4 3 4 4 Overview 4 4 1 Conventions General System Settings Installing the Plate Sample Applications 4 4 2 Example 1 Injecting a Single 200 nL Sample from 1 vial 4 4 3 Example 2 Injecting a Single 100 nL sample from 3 vials and Injecting 5 uL sample from 3 other vials 4 4 4 Example 3 1uL injection out of a 5 uL sample i
107. l at various temperature setpoints and set the cooling option so that the desired intemal temperature is attained This key is used to access a variety of service related commands and it is protected by a code Use of these commands is restricted to authorized service personnel D931R1 User s Manual FAMOS The User Interface 3 8 2 The System Menu When you press the System key the display asks whether the autosampler should operate in the lt MICRO gt or in the lt CONVENTIONAL gt mode lt MICRO gt lt CONVENTIONAL gt The ranges and the default settings of some system parameters are dependent on this selection of the mode of operation IES Note If you change from one operating mode to the other all parameters will be set to their default values for that mode In both operating modes the System Menu offers the soft function keys presented in FIGURE 3 4 and which are discussed in detail in Sections 3 8 3 3 8 8 lt CLOCK gt and lt COMM gt are available after pressing the Menu key lt GENERAL gt lt USAGE gt lt PLATES gt lt 10 gt lt CLOCK gt lt COMM gt e Loop volume e Method protection e Type of plate e Needle tubing code e Process in e Syringe volume 1 e Use time based ROWS COLUMNS e Syringe speed method e Number of transport e Needle height e Use mix method vial e Skip missing wells e Use user program e Inj Marker pulse e Well marker pulse e Lab Well marker pulse e Inp
108. ld perform a wash sequence in which all tubing is rinsed and the valve and I O ports are reset At the end of the sequence serial mode operation is resumed Press to end serial mode and return to the Ready Menu This soft key is used to start a procedure in which all lines of the solvent selection valve can be primed if this option is included in the system This soft key is used to enter the programming mode for Peltier plate cooling if this option is included in the system If the cool option is switched lt ON gt the following soft function keys can be selected The temperature control will remain OFF until it is switched on again by the user in this menu The temperature control will be switched OFF after all programmed series have been executed The temperature control will be switched OFF at a date and time that can be user selected eS Note The programmable temperature range is 4 C to 40 C The maximum cooling capacity is approximately 20 C below ambient Make certain that the condensed water and leakage tubes are connect to a waste container on the floor to drain condensed water ae Note The temperature of the sample inside the sample vial may be slightly different than the temperature set due to a variety of effects e g the heat transfer characteristics of the vial walls If it is necessary to precisely set the temperature of the sample we recommend that you determine the temperature inside the via
109. le needle arm did not reach or leave Contact Reena destination within a certain time horizontal LC Packings ERROR 34 Sample needle arm not in vertical home position Contact eee while moving horizontally LC Packings ERROR 39 Vial sensor sticks Contact ee eee Le packings ERROR 40 The sample needle spindle does not rotate Contact ERROR ore mel s ERROR 41 The sample needle did not reach or leave home Contact S E nee a ERROR 42 The sample needle is not at home position Contact E e ERROR 53 The sample needle arm is not in the home Contact ERROR 8 er e Packings TABLE 6 10 Error Codes for Vials Probable Cause Solution ERROR 60 Missing vial Only available when Skip Missing Check install the Vial is set to NO in the System Settings and vial during the execution of the Mix of a sample on 48 vial plate ERROR 62 Missing transport vial Check install the AAA E vial ERROR 65 Missing vial for reagent B Check install the vial ERROR 66 Missing vial for reagent C Check install the vial ERROR 67 Missing vial for reagent D Check install the vial ERROR 68 Missing destination vial Check install the vial ERROR 69 Not enough transport liquid available due to Check install the missing transport vials vial TABLE 6 11 Error Codes for Electronics Probable Cause Solution ERROR 71 Flex PCB of the sample needle is not properly Contact ERROR a TA e ERROR 72 Invalid configuration of the FAMOS Well Plate Contact ERRO
110. led in System Menu Usage Menu lt EDIT gt lt INSERT gt lt DELETE gt lt ASPIRATE gt lt DISPENSE gt This soft function key is used to access a Series of commands to program a method for e pre injection sample handling e g pre column derivatization e dilution e adding of internal standard Up to nine mix methods can be programmed A total of 240 steps can be included in the 9 mix methods and the user program Assign a number to the mix method When the Mix Menu Is presented Press to edit an existing step or a new step for a new mix method Press to insert a new step in an existing method before the displayed step Press to delete the displayed step eS Note End of mix method means that the mix method is empty if an existing mix method is selected the first line of the mix method is displayed Scroll through the steps of the existing method with the cursor keys and use the soft function keys to enter changes in an existing method The following types of steps can be programmed for a mix method Aspirates a programmed volume sample air destination reagent A D Speed of syringe can be selected from 1 9 TABLE 3 1 The indicated height H is the distance of the needle point to the plate holder default 2 mm The maximum amount which can be aspirated is the total volume of the syringe Dispenses Sample waste destination reagent A D a programmed volume from the buffer
111. leshooting This page intentionally left blank 6 16 D931R1 User s Manual FAMOS CHAPTER 7 Specifications 7 1 General 7 1 1 Analytical Performance Capped and Reproducibility sealed vials partial loopfill RSD lt 0 6 injection volumes 1 uL with headspace pressure on the vial and 5 wL pre flush RSD lt 1 5 injection volumes 1 wL without injections headspace pressure on the vial Open vials plates Reproducibility n 1 0 cP RSD lt 1 0 injection volumes 1 uL without injections headspace pressure on the well RSD lt 1 5 injection volumes 1 uL without injections headspace pressure on the vial Memory effect lt 0 01 with programmable needle wash User s Manual FAMOS D931R1 7 1 Specifications 7 1 2 Programming Full loop injections Injection methods Partial loopfill injections uL pick up injections 0 01 uL 1000 uL full loop depending on system settings 0 01 500 uL with 0 1 uL increments for partial loopfill Injection volume 0 1 uL max volume with 0 01 uL increments for uL pick up max volume loop volume 3 x needle volume 2 7 1 3 Inputs Outputs 2 RS 232 Digital Output 2 EVENT Open Collector Relay START ERROR IN Optocoupler START ERROR OUT Open Collector Analog Input 1 V 1V 100 nm Analog Output 2 0 1 V 1 V 10 V for recorder or integrator 7 1 4 Physical Pimen ong 280 mm 11 0 in x 400 mm 15 7 in x
112. lume to 10 uL and verify that the right needle height 4 mm has been set so that the needle can reach the bottom of the tapered sample vial see Section 4 2 d Identify the position of the transport liquid vial by entering the parameters indicated in TABLE 4 9 TABLE 4 9 Identifying the Location of the Transport Vial System to enter the system settings lt MICRO gt E to select micro mode lt PLATES gt E to select the right plate version lt 48 gt E lt IN ROWS gt to process the well in rows 01 E to define the location of the first transport liquid vial 01 E to define the location of the last transport liquid vial Escape Escape to return to the Ready M enu Users Manual FAMOS D931R1 4 7 Programming Examples e Program the Injection Method by entering the parameters indicated in TABLE 4 10 TABLE 4 10 Programming the Injection Method Methods to enter the Methods Menu lt INJ ECTION gt 01 E program injection method number 1 lt PICK UP gt E to select partial loopfill injection method 25 00 E to define an analysis time of 25 minutes 1 E to define the number of injections vial 1 00 E to set the injection volume to 1 00 uL Escape Escape to return to the Ready Menu f Define the Series by entering the parameters indicated in TABLE 4 11 TABLE 4 11 Programming the Series Series to enter the Series Menu 01 E to define the Series number 01 E to select the injection method number CLE to
113. lumn oven User s Manual FAMOS D931R1 7 3 Specifications 7 1 8 Plate Dimensions Whos eee Als 4 gt 103 5 N 3 C Titerplate 384 J 4 29311 User s Manual FAMOS APPENDIX A Injection Principles This appendix describes detailed information about the modes of injection used by the FAMOS Well Plate Microautosampler A 1 Full Loop Injections A 1 1 Standard Mode The switching sequence for a full loop injection is described in this section At the start of the sequence the injector is in the INJECT position FIGURE A 1 and the sample needle has entered the well after the air needle has pre punctured the septum Headspace pressure is applied through the outer air needle to ensure that no air or vapor bubbles are formed during sample aspiration lt Syringe gt FIGURE A 1 Injector is in the INJECT Position The syringe dispenser aspirates the programmed flush volume from the sample well to fill the sample line with sample and remove wash solvents FIGURE A 2 User s Manual FAMOS D931R1 A 1 Injection Principles
114. method number 3 CL E to select lt NONE gt for wash method lt ROW A gt 1 E to define location of first sample well lt ROW A gt 1 E to define location of last sample well lt ROW B gt 1 E to define location of first destination well 1 E to define position 1 for the Reagent A vial Escape to return to the Ready Menu f Run the series by entering the parameters indicated in TABLE C 13 and pressing lt START gt TABLE C 13 Programming System Parameters Start Stop to start the FAMOS Well Plate 01 E to start at series number 1 01 E to stop after series number 1 lt START gt to start processing of sample The FAMOS Well Plate will now start searching for the Reagent vial and transport 90 uL to the destination well once then 1 uL of sample will be added and after mixing 3 times a 1 uL injection will be performed C 8 D931R1 User s Manual FAMOS Additional Programming Examples C 5 Defining a Template and Adding a Protection Code This example describes how to incorporate an injection method 02 and a wash method 01 which were defined in Section C 3 into a template A protection code will be added After use of templates has been enabled the message ALL SERIES DEFAULT appears The user will have to redefine the series because the settings have been changed a To select the methods to be incorporated in the template enter the parameters described in TABLE C 14 TABLE C 14 Programming th
115. mpletely quantitatively filled with sample This results in extremely good reproducibility e Partial loopfill The sample loop is partially filled with sample This provides low sample loss The operator can select the desired injection volume e pL pick up After aspiration of sample the sample is transported into the loop with transport liquid mobile phase In this mode no sample is lost Technical information on the injection principle used by the FAMOS Well Plate Microautosampler is presented in Appendix A 1 2 D931R1 User s Manual FAMOS System Overview 1 3 General Design of the FAMOS Well Plate Microautosampler The front view of the FAMOS Well Plate Microautosampler is presented in FIGURE 1 2 Cover in open position Buffer tubing Injection valve including the Sample Loop 3a Tubing holder Needle unit Screw to fasten cover Transport reagent vials Plate Plate holder 10 Drain wash position 11 Condensed water and leakage 12 Wash position 13 Wash solvent bottle 14 Syringe 15 Syringe waste tubing OONOOABWDNML gt FIGURE 1 2 Front View of the FAMOS Well Plate Microautosampler User s Manual FAMOS D931R1 1 3 System Overview 1 4 Contents of this Manual IIe Note This manual covers the standard version of the FA
116. ms c A purchase order if the system is not in warranty User s Manual FAMOS D931R1 vii Instructions for Returning Instruments This page intentionally left blank viii D931R1 User s Manual FAMOS Warnings The Danger sign Warning sign and the Hazard sign shown below are included in various locations in this manual These signs provide the following information gt DANGER Danger The information in a danger statement relates to a procedure practice condition or action that if not done correctly or adhered to could lead to personal injury or loss of life gt WARNING Warning The information in a warning statement relates to a procedure practice condition or action that if not done correctly or adhered to could lead to severe injury and or damage or destruction to parts or all of the equipment gt CAUTION Caution The information in a caution statement relates to a condition that could lead to damage to equipment and or lead to invalid analytical results 4 Note The information in a note statement relates to important information that should be read and understood before continuing Safety Precautions IE Note The following precautions should be followed to minimize the possibility of personal injury and or damage to property IE Note Make certain that you are familiar with the contents of this manual before working on the system 1 The system should be installed in a well ventilated laboratory
117. n vial A1 using the transport liquid uL pick up Testing the Microautosampler 5 1 O22 5 3 5 4 Overview Establishing a Test Protocol The Test Protocol 5 3 1 The Analytical System used for the Test Protocol 5 3 2 The Test Protocol Checking System Components 5 4 1 Testing the Accuracy of Delivery of the Syringe 5 4 2 Testing the Loop Volume 5 4 3 Testing the Tray Cooling Option Maintenance and Troubleshooting 6 1 6 2 6 3 Overview Maintenance Replacing Major Components 6 3 1 Replacing the Syringe 6 3 2 Replacing the Syringe Tip 6 3 3 Needle Assembly 6 3 4 Prepuncturing Needle 6 3 5 Combination Syringe Sample Loop and Buffer Tubing 3 8 3 10 3 10 3 13 3 13 3 15 3 16 3 16 3 17 3 17 3 17 3 18 3 19 3 20 3 21 3 22 3 25 3 26 5 7 5 1 5 2 5 3 5 3 5 3 5 5 5 5 5 5 5 6 6 7 6 1 6 2 6 3 6 3 6 4 6 4 6 6 6 7 D931R1 User s Manual FAMOS Table of Contents CHAPTER 7 APPENDIX A APPENDIX B APPENDIX C APPENDIX D 6 3 6 Replacing the Main Fuses and Setting the Operation Voltage 6 4 6 5 6 6 6 6 7 Major Items 6 6 2 Needle Assembly 6 6 3 Syringe and Buffer Tubing 6 6 4 Injection Valve Troubleshooting Error Codes of the Microautosampler Spare Parts List 6 6 5 Vials and Accessories Specifications 7 1 NNN NN NN N General 1 1 Analytical Performance 1 2 Programming 1 3 Inputs Outputs 1 4 Physical 1 5 Electrical 1 6
118. ne a x Service engineer Address Telephone Fax Comments User s Manual FAMOS D931R1 F 9 Programming Forms This page intentionally left blank F 10 D931R1 User s Manual FAMOS APPENDIX Connecting the NANO Column or other Fused Silica Capillary G 1 Introduction These directions should be used when a NANO column or a fused silica capillary is to be connected to the inert PAEK injection valve of the FAMOS Micro autosampler or the inert PAEK switching valves of the Switchos II Advanced Microcolumn Switching Unit CAUTION Caution Do not use a stainless steel nut and or ferrule with inert PAEK injection switching valves The use of stainless steel nuts or ferrules may damage the valve Use only the supplied fittings PEEK and follow the instructions below G 2 Using PEEK Fingertights The PEEK sleeve connection is created using the components shown in FIGURE G 1 e PEEK fingertight nut e PEEK ferrule e PEEK sleeve with an appropriate D e NANO column or any other fused silica capillary e The female fitting port of the inert valve FIGURE G 1 shows a stainless steel union which is used only for the assembly of the connection Union Sleeve Fingertight Nut IC PACKINGS _ ap pap m CES MAHI Ferrule NANO Columnn FIGURE G 1 Parts of a PEEK sleeve connection User s Manual FAMOS D931R1 G 1 Connecting the Nano Column G 3 Fitting
119. nse action If you want the needle to stay in the same position an aspirate or dispense step of 0 uL must be programmed at the desired position This command is used to activate the compressor to put air pressure on a sample The compressor will stay active lt COMPRES gt 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 This command is used to control the four standard lt AUX gt auxiliaries contact closures Refer to Appendix E for details This command is used to program a pause in which the FAMOS Well Plate Microautosampler waits for one of the lt WAIT IN gt four inputs to become lt HIGH gt or lt LOW gt before continuing with the next step Refer to Appendix E for details This command is used to define two programmable outputs contact closures These are similar to the auxiliaries but only available in the user program Refer to Appendix E for details lt PROG OUT gt to program the output to the connector P3 TIMED lt CODE gt OUTPUTS This is a HEX output in the range 0 to 15 Refer to Appendix E for details the markers normally generated in the FAMOS Well Plate Microautosampler are not active in the user program but lt MARKERS gt can be programmed in this screen refer to Appendix E for details Select marker and status inject vial labeled lt SSV gt This command is us
120. nt temperature is below 18 C 5 3 2 The Test Protocol Prepare the FAMOS Well Plate Microautosampler as follows e Place a closed vial at position A01 the sample is Uracil in distilled water 0 1 mg mL The eluent is distilled water e Fill the wash solvent bottle with 80 H2O 20 isopropanol e Program a partial loop fill method using the settings in Section 5 2 Obtain the peak area for each peak and determine the value for o as well as the RSD reproducibility which should be lt 1 0 using the equations presented below _________ Yt Peak area Peak area _____ n gt Peak area Peak area o SS T 3 n 1 RSD x100 Peak area User s Manual FAMOS D931R1 5 3 Testing The Microautosampler A typical chromatogram from the test is presented in FIGURE 5 2 and sample data is presented in TABLE 5 4 Mg Chromatogram Viewer O File Setup View Data Integration Report Options Help Time miri FIGURE 5 2 Typical Sample Chromatogram for the reproducibility test TABLE 5 4 Sample Peak Height Data Peak number Height 1 2 3 4 5 6 7 8 9 For this example the results are RSD 0 66 The specified value is an RSD lt 1 If the reproducibility is not within the specification a Check for any air bubbles in the tubing syringe b Check needle and tube connections between needle and injection valve for dead volume or blockage c Review the Troubleshooting Sect
121. o fill the wash solvent tube you may have to repeat the flushing procedure a few times 2 5 4 Flushing the Syringe The FAMOS Well Plate Microautosampler is supplied with a 25 wL syringe It is also possible to use the microautosampler with a 100 uL 500 uL 250 ul or 1000 uL syringe Refer to Section 6 3 1 for instructions how to replace the syringe 2 10 D931R1 User s Manual FAMOS Installation and Getting Started To remove the air and flush the syringe a Check the general settings in the System Menu Section 3 8 and make sure that the appropriate syringe and the corresponding buffer tubing is installed b Select soft function key lt WASH gt in the Ready Menu to perform a standard wash routine All tubing connected to the syringe valve is filled and rinsed c If any air remains in the syringe perform an additional wash cycle and gently tap the syringe as wash solvent is dispensed to waste ae Note If there is still air in the syringe gently tap the syringe as wash solvent is dispensed to waste during the wash cycle 2 5 5 Adjusting the Needle Height The sample needle height can be programmed within the general system settings used for injection methods only the mixed methods and within user defined programs Chapter 3 This parameter is defined as the distance from the top of the plate holder to the bottom of the sample needle when the sample needle is moved all the way down This parameter can be adju
122. ogramming examples a discussion about the injection valve and the various inputs and outputs on the rear panel If you are using the FAMOS Well Plate Microautosampler with the LC Packings UltiMate and or U tiChrom software please refer to the documentation provided with these products for supplemental information If the microautosampler is used with other systems the manuals provided with these systems should be consulted for interfacing requirements 1 4 D931R1 User s Manual FAMOS in Installation and Getting Started 2 1 Installation The instructions provided below are provided for installation of the FAMOS Well Plate Microautosampler as a stand alone component in an HPLC system When the FAMOS Well Plate Microautosampler is used in conjunction with a LC Packings UltiMate system please refer to the U tiMate User s Manual for additional information Once you have set up the microautosampler refer to Section 2 6 for information about routine operation of the system 2 1 1 Location of the FAMOS Well Plate Microautosampler in the Laboratory The FAMOS Well Plate Microautosampler should be installed in a facility with the following environmental conditions e The temperature range should be maintained between 10 and 40 C The system should be installed in an area in which the temperature is fairly constant do not place the system near a window an air conditioning duct or a heating duct The humidity should be maint
123. on method number wash method number timebase method number A maximum of 24 templates can be programmed Users Manual FAMOS D931R1 3 25 The User Interface 3 9 Series Menu The Series Menu allows the user to define the run sequence in a series A maximum of 24 series can be programmed each series contains information about the methods to be used for a range of wells This can be a template a separate method mix injection wash timebase or the user program Information on location of wells labeled wells or calibration wells is also programmed in a series Series parameters are described in TABLE 3 4 eS Note The settings entered in the System Menu and the methods defined in the Methods Menu determine which possibilities appear in the Senes Menu TABLE 3 4 Series parameters Without templates With templates O Use user program Yes No Template number Injection method number Wash method number OTime base methods number O Mix method number Time base and mix method are only available if enabled in the System Menu O Use calibration wells Yes No O First calibration well O Last calibration well O No of samples between calibration Calibration wells are only available if enabled in the System Menu not available if Mix Method has been programmed First sample well Last sample well Only if a mix method has been programmed or if used in user program O First destination well O Vial Reagent A O Vi
124. on well lt DESTINATION gt lt INSERT gt to define mix method step number 11 lt WAIT gt 02 E to wait 2s lt INSERT gt to define mix method step number 12 lt DISPENSE gt 30 E to dispense 3 0 uL to waste lt WASTE gt WASH 300 E to perform a wash with 300 uL lt INSERT gt to define mix method step number 14 lt ASPIRATE gt 20 E to aspirate 2 0 uL air lt AIR gt lt INSERT gt to define mix method step number 15 lt ASPIRATE gt 50 to aspirate 5 0 uL from the destination well lt DESTINATION gt note step will improve lt INSERT gt to define mix method step number 16 lt DISPENSE gt 50 to dispense 5 0 uL to the destination well lt DESTINATION gt step 15 and 16 are used to mix the destination lt INSERT gt to define mix method step number 17 lt REPEAT gt 2 4 2 E to repeat last 2 steps 2 times lt INSERT gt to define mix method step number 18 lt DISPENSE gt 20 E to dispense 2 0 uL to waste lt WASTE gt lt INSERT gt lt MENU gt to define mix method step number 19 lt WASH gt 300 E to perform a wash with 300 uL User s Manual FAMOS D931R1 C 7 Additional Programming Examples e Program the series by entering the parameters indicated in TABLE C 12 TABLE C 12 Programming Series Parameters Series to enter the Series Menu 01 E to define series number 1 01 E to select Mix method number 1 for this series 03 E to select Injection
125. optimal results with any of the injection principles used in the FAMOS Well Plate Microautosampler including the partial filling and the uL Pick Up methods in which the injection volume is determined by the syringe and full loop injection in which the volume is determined by the size of the loop The design prevents any contact between the needle and the stator and rotor faces Ie Note A detailed discussion on the Installation Use and Maintenance of the valve is presented in Technical Note 801 from Valco Instruments Co Inc and can be obtained at the Valco website www Valco com User s Manual FAMOS D931R1 D 1 The Sample Injector D 2 Maintenance In most instances the only maintenance that is required is cleaning of the valve Cleaning can often be accomplished by flushing all the lines with an appropriate solvent s The selection of the solvent is dependent on the nature of the sample and the mobile phases that are used Typically solvents such as methanol acetonitrile methanol water 80 20 or acetonitrile water 80 20 should be used D 3 Disassembly Reassembly of the Valve Re Note Do not disassemble the valve unless system malfunction is definitely isolated to the valve D 3 1 Disassembly of the Valve To disassemble the valve a Use a 9 16 hex driver to remove the socket head screws which secure the cap to the valve FIGURE D 1 Driver Rotor Stator FIGURE D 1 Exploded View of the Valco
126. options available for the FAMOS Well Plate Microautosampler Note When the injection valve is mounted on the right side e g using the FAMOS Well Plate Microautosampler in combination with the THERMOS the sample needle should be an extra long version 6 3 4 Prepuncturing Needle To replace the prepuncturing needle a Remove sample needle Section 6 3 3 b Unscrew the prepuncturing needle c Install a new needle using a new seal d Reinstall the sample needle Section 6 3 3 6 6 D931R1 User s Manual FAMOS Maintenance and Troubleshooting 6 3 5 Combination Syringe Sample Loop and Buffer Tubing This section will indicate the standard configuration of the FAMOS Well Plate Microautosampler and appropriate configuration for three commonly selected modes of using the microautosampler a detailed discussion of the injection principles is presented in Appendix A Injection volumes smaller than 1 uL Injection volumes up to twice the standard conventional mode For volumes larger than 200 uL A Standard Configuration The standard configuration for the FAMOS Well Plate Microautosampler includes a 2 4 uL Fused Silica Needle a 25 uL syringe a 50 uL buffer tubing and a 10 wL sample loop The injection volume ranges shown in TABLE 6 4 are available for the various injection modes while TABLE 6 5 presents the maximum injection volumes for the various injection modes Five sizes of syringes can be used in the FAMO
127. ponents due to wear or to re configure the system e g changing the syringe to meet the requirements of a different analytical procedure Section 6 3 Troubleshooting discusses a series of activities that should be used to determine the cause of a problem Section 6 4 Error Codes lists and describes various messages that are presented on the display to indicate a fault with the system Section 6 5 Spare Parts Replacement Parts Lists Presents a listing of components that are used to maintain the unit or to change the configuration Section 6 6 User s Manual FAMOS D931R1 6 1 Maintenance and Troubleshooting 6 2 Maintenance Maintenance refers to a variety of activities that should be performed on a routine basis to optimize performance of the system Many routine maintenance activities can be readily performed by the user In some cases e g replacement of critical components we recommend that a factory trained service engineer should be called to perform the operation This will ensure optimal long term performance and maximum uptime LC Packings provides a broad range of service support activities to ensure that the FAMOS Well Plate Microautosampler is functioning in a suitable manner These activities can be customized to meet the specific needs of the customer For further information please contact your local LC Packings office or representative TABLE 6 1 Recommended Maintenance Schedule Frequency
128. r repair IIe Note When re installing the valve make certain that the proper tubing is attached to the appropriate fitting The loop is connected to ports 1 and 4 the pump is connected at port 2 and the column is connected at port 3 User s Manual FAMOS D931R1 D 3 The Sample Injector This page intentionally left blank D 4 D931R1 User s Manual FAMOS APPENDIX E Digital Inputs and Outputs E 1 Introduction The rear panel of the FAMOS Well Plate Microautosampler includes a series of I O connectors that allow the user to interface the microautosampler to other components in their analytical system If the microautosampler is used with a LC Packings U tiMate Capillary and Nano HPLC systems refer to the User s Manual for that system for specific installation interfacing information User s Manual FAMOS D931R1 E 1 Digital Inputs and Outputs E 2 Contact Closure Outputs I O Connector P1 programmable outputs P4 marker outputs and P5 auxiliary outputs are contact closure outputs floating NO NC contact as shown in FIGURE E 1 Normally Closed Common C 4 COM O FIGURE E 1 Contact Closure Output E 2 1 P1 Connector OUTPUTS TABLE E 1 P1 Connector Pin Descriptions Normally open Common Normally closed Normally open OUT 2 Common Normally closed 24 V DC OUT 1 Normally Closed pagar C NC COM O Normally Open NO NC Normally Open
129. rt the FAMOS Well Plate Microautosampler again caution Caution A panic stop does not remove the sample from the buffer tubing Before starting again you should perform a wash sequence After the FAMOS Well Plate Microautosampler has completed the run the Ready Menu will appear again A number of examples of the use of a series are presented in Chapter 4 and Appendix C It is possible to program a series and or a method during a run by pressing Series or Methods The menus that are presented in this mode are identical to those offered when the FAMOS Well Plate Microautosampler is idle 3 8 D931R1 User s Manual FAMOS The User Interface If a series or method is changed the new values become active the next time the FAMOS Well Plate Microautosampler initiates a series The series currently running are not affected by the changes 3 7 2 Executing a Series via Remote Control After the range of series has been programmed the FAMOS Well Plate Microautosampler will ask for the Start conditions If you press the lt REMOTE gt soft key the Remote mode of operation will be selected In this mode the autosampler will act as a slave of another device e g an HPLC pump and can be controlled with the NEXT INJ ECTION and NEXT VIAL inputs of connector P6 see Appendix E e NEXT INJ ECTION INPUT this input will start the next injection of the series If the series was completed it starts the next injection from the next series
130. ry Installed Items lem i escription O Wash Solvent Bottle 100 mL Fuses 115 V AC 10 IN power switch two 5 AT fuses slow 4 x 1 UL CSA 230 V AC 10 two 2 5 AT fuses slow 5 x 20 mm IEC127 The fuses used are UL listed and CSA certified TABLE 2 2 Standard Tubing Configuration Material and Dimensions and tubing 0 100 mm I D total volume 2 4 uL Buffer tubing from high PTFE tubing 255 mm x 1 16 O D x 0 5 mm I D pressure valve to syringe volume 5O yL valve Tubing syringe valve to PTFE tubing 300 mm x 1 16 O D x 1 0 mm I D wash solvent bottle Tubing syringe valve to PTFE tubing 400 mm x 1 8 O D x 1 6 mm I D waste User s Manual FAMOS D931R1 2 3 Installation and Getting Started 1 Cover in open position 2 Buffer tubing 3 Injection valve including the Sample Loop 3a 4 Tubing holder 5 Needle unit with Sensor 5a 6 Screw to fasten cover 7 Transport reagent vials 8 Plate 9 Plate holder 10 Drain wash position 11 Condensed water and leakage 12 Wash position 13 Wash solvent bottle 14 Syringe 15 Syringe waste tubing FIGURE 2 1 Front View of the FAMOS Well Plate Microautosampler 2 4 D931R1 User s Manual FAMOS Installation and Getting Started CAU
131. s full loop injection flush volume 30 uL number of injections per vial 1 e Program Time Base 01 with the parameters shown in TABLE C 20 TABLE C 20 Programming Time Base ISS A 1 6 00 45 switches pre column on line after sampler loading switches the ISS A valve back and the ISS A 1 2 14 50 pre column off line again for the next sample AUX 1 00 45 starts data acquisition of the data system AUX 1 00 46 e Program Series 01 using Injection Method 01 and Time Base Method O1 mw e Start the FAMOS Well Plate Microautosampler IES Note The above example shows how to perform column switching with the FAMOS Well Plate Microautosampler equipped with one ISS valve For advanced column switching applications e g on line digestion of proteins on line removal of non ionic detergents from protein peptide samples LC Packings offers the SWITCHOS equipped with two valves and the Switchos Il equipped additionally with a Loading Pump and Solvent Selection Valve Advanced Micro Column Switching Units Please contact LC Packings for additional information User s Manual FAMOS D931R1 C 13 Additional Programming Examples This page intentionally left blank C 14 D931R1 User s Manual FAMOS APPENDIX D The Sample Injector D 1 Overview The Cheminert Model C2 Injection Valve is a 6 port external loop injector manufactured by Valco Instruments Co Inc It is designed to offer
132. s in this appendix are provided to allow the user to get an overview of the various commands tn each menu and how the various commands are accessed The FAMOS Microautosampler D931R1 B 1 Programming Charts Ready Menu lt Menu gt lt PLATES gt lt WASH gt lt SYR END gt lt UTILS gt lt SERIAL gt lt SSV gt lt COOL gt lt SERVICE gt lt COPY lt ERASE gt lt LOG gt lt DEFAULT ALL gt Exchange Perform Move syringe Copy method Erase methods Logbook All methods Communication Prime solvent Service mode Lift_left standard wash amp Userprog erased all settings protocol valve Lift_right to default Plate System E System Settings autosampler lt MICRO gt lt CONVENTIONAL gt SystemSettings lt Menu gt lt GENERAL gt lt USAGE gt lt PLATES gt lt lO gt lt CLOCK gt lt COMM gt e Loop volume Method protection code Je Type of plate Inj Marker pulse e Device identifier e Needle tubing Use time based method e Process in Well marker pulse e Syringe volume 1 Use mix method ROWS COLUMNS Lab Well marker pulse e Syringe speed Use user program e Number of transport Input edge next inj e Needle height Use labeled wells vial Input edge next well e Skip missing wells Use templates Freeze input active e Air segment Use calibration wells Reset outputs after e Headspace pressure series e Time base display e Key click e Error buzzer e Alarm buzzer 1 only in Conventional mode
133. ser Program 3 22 lt AUTOMATIC gt 3 12 lt AUX gt Timebase 3 20 User Program 3 24 Auxiliaries P5 Connector 2 6 B Buffer Tubing 2 3 2 4 6 7 C lt CALIBRATION WELLS gt 3 15 CE mark 2 5 CE Declaration xi Checking Loop Volume 5 5 Syringe Accuracy 5 5 Tray Cooling Option 5 6 CL Clear key 3 3 lt CLOCK gt 3 17 lt CODE gt Timebase 3 20 User Program 3 24 Column Switching C 12 lt COMM gt 3 17 Communication Connector 2 6 lt COMPRES gt 3 24 Condensed water and leakage 2 9 Configuration Standard 2 3 Injection lt 1 uL 6 7 Injection gt 200 uL 6 8 Standard Injection 6 8 Connecting to an HPLC System 2 8 Connections Electrical 2 5 Auxiliaries P5 2 6 E 3 Communication 2 6 Markers P4 2 6 E 2 Outputs E 2 Power 2 7 Contact Closure E 2 Contents of Manual 1 4 lt COOL gt 3 12 Cooling 3 12 Testing 5 6 lt CONVENTIONAL gt 3 13 lt COPY gt 3 11 lt COUNTS gt 3 11 Cursor Keys see Arrow Keys 3 3 D lt DATE TIME gt 3 12 lt DEFAULT ALL gt 3 11 lt DELETE gt Mix Method 3 21 User Program 3 22 lt DESTINATION gt C 7 Device Identifier COMM Menu 3 17 Digital Inputs and Outputs E 1 lt DISPENSE gt Mix Method 3 21 User Program 3 23 Display 3 3 Drain Wash position 2 9 E E Enter key 3 3 lt EDIT gt Mix Method 3 21 User Program 3 22 END TIME lt 3 20 gt lt ERASE gt 3 11 Electrical Connections 2 5 Entering a Program 3 6 lt ERROR BEEP gt 3 14 Error Codes 6 11 Es
134. sted to insert the sample needle as far as possible into the sample vial to aspirate the whole sample out of the vial FIGURE 2 9 The default setting is 2 mm Prepuncturing Needle Sample Needle 16 mm Sample Vial 14 mm 12 mm 10 mm Well Plate 8mm z 6 mm 3 b 4 mm Z Y Plate Holder aoe 0 mm __ FIGURE 2 9 Positioning the Sample Needle CAUTION Caution A sample needle height O corresponds to the top of the plate holder Operating the FAMOS Well Plate Microautosampler with this value set to O and with a well plate installed at the same time may damage the sample needle This is especially true if the conventional stainless steel needle is used User s Manual FAMOS D931R1 2 11 Installation and Getting Started 2 6 Routine Operation of the System 2 6 1 Sample and Mobile Phase Considerations The FAMOS Well Plate Microautosampler is used in an HPLC system and the standard operating precautions for HPLC should be employed e Ensure that samples and mobile phases do not contain particulate matter All samples and mobile phases should be filtered through a 0 22 um membrane filter If organic solvents are used make sure that extractable materials are not present in the filter e The sample should be soluble in the mobile phase If a gradient is used make certain that the sample is soluble in the mobile phase at all mobile phase compositions to be used in the separation e After you have finis
135. t the values in TABLE 5 1 for the General Parameters TABLE 5 1 General Parameters Air segment NOU b Set the well plate type you are using c Program the Partial Loopfill method presented in TABLE 5 2 TABLE 5 2 Method Parameters Injections vial 9 d Program the Series presented in TABLE 5 3 TABLE 5 3 Series Parameters Series number 1 Injection method number Wash method ROW A01 ROW A01 5 2 D931R1 User s Manual FAMOS Testing The Microautosampler 5 3 The Test Protocol 5 3 1 The Analytical System used for the Test Protocol In this test protocol a LC Packings U tiMate Micropump with a CAP 300 calibrator and an U tiMate UV Detector set at 254 nm are used FIGURE 5 1 The U tiChrom software is used to control the system and acquire present the data A 1 m fused silica tube 75 um I D should be placed between the autosampler and the detector to create backpressure Ultimate capillary LC system FAMOS Well Plate L autosampler Frm 1 m fused silica tubing r um 1 0 FIGURE 5 1 Analytical System for Test Protocol If an U tiMate system is not available any HPLC system with reproducible microflow capabilities can be used e g a conventional HPLC pump with an Acurate flow splitter I Note The entire flow path from the syringe to the needle should be carefully inspected before the test protocol is performed This test should not be performed if the ambie
136. t this input is active If the FREEZE INPUT is activated while the analysis time is not running the FAMOS Well Plate Microautosampler will perform all programmed pre injection sample handling mix method and loading part of the injection method It should be noted that the FAMOS Well Plate Microautosampler will not inject the sample until the FREEZE INPUT is no longer active Stop I O With this input the run of the FAMOS Well Plate Microautosampler is immediately aborted and the Ready Menu appears in the display If the FAMOS Well Plate Microautosampler is in remote control the run of the FAMOS Well Plate Microautosampler is immediately aborted but it remains in remote control and cannot be restarted via a NEXT INJECTION input INPUT 1 4 Programmable inputs can be used in the user program E 6 D931R1 User s Manual FAMOS AEREN Programming Forms The powerful programming features described in Chapter 3 allow the user to set sampling parameters to meet the precise needs of the analyst On the following pages we provide blank sheets which can be used for hard copy records of System Menu Settings Templates Injection Methods Wash Methods Timebase Methods Mix Methods and User Programs In addition we provide a form which can be used to describe the FAMOS Well Plate Microautosampler This latter form should be faxed to LC Packings when you are enquiring about your system as it includes information that may assist in identify
137. the FAMOS Well Plate Microautosampler The FAMOS Well Plate Microautosampler is a flexible and powerful system for micro HPLC and incorporates the following features e It can be used with a broad range of plates from various manufacturers 96 well plate 96 deep well plate 384 well plate 48 1 5 mL autosampler vials sealed or open e It is capable of executing full loop injections partial loopfill injections and uL pickup e It supports Low Dispersion injection control to optimize injection profiles e It can use a broad range of syringes volumes of 25 100 250 500 or 1000 uL e It is fully controlled by U tiChrom software e It is fully compatible with other laboratory equipment e t can be used for both routine analysis and method development purposes e t is menu driven and includes a broad variety of sample handling programs such as sampling diluting mixing and precolumn derivatization A number of options e g Peltier plate cooling and a column oven are available for the FAMOS Well Plate Microautosampler In addition a broad range of components to configure the unit to meet your requirements e g loops injection needles syringes needle holder for nanospray and disposable items e g vials and caps are available A detailed list of available items is presented in Section 6 6 User s Manual FAMOS D931R1 1 1 System Overview 1 2 Principle of Operation of the Microautosampler A schematic
138. to define the type of plates that will be used Four types can be selected 96 low default 96 high 384 low or 48 vials After a plate type has been selected press ENTER WELL PROCESSING Indicates if samples should be processed in rows left to METHOD right or in columns top to bottom TYPE OF PLATE FIRST TRANSPORT Enter a number 1 4 or press CL VIAL LAST TRANSPORT Enter a number 1 4 VIAL eS Note Vials can be placed in any of the four positions Transport vials must be placed ina continuous row 3 8 6 The System Menu Item lt lO gt This soft function key Is used to access the IO configuration mode and define the following the default and range are indicated in TABLE 3 2 INJ ECT MARKER This entry is used to define the length of the inject PULSE LENGTH marker pulse This entry is used to define the length of the well marker WELL MARKER pulse PULSE LENGTH LABELED WELL This entry is used to define the length of the well marker MARKER PULSE pulse of the labeled well LENGTH INPUT EDGE NEXT This entry is used to define the edge sensitive inputs for INJ ECTION the next injection FREEZE INPUT This entry is used to define whether the freeze input is ACTIVE active when high or freeze input is active when low RESET OUTPUTS This entry is used to indicate whether the outputs should AFTER LAST SERIES be reset to default after the last series eS Note Refer to Appendix E for
139. tubing Speed of the syringe can be selected from 1 9 TABLE 3 1 The indicated height H indicated is the distance of the needle point to the plate holder default 2 mm It Is possible to dispense a larger volume than the volume aspirated in previous actions The aspirated amount will be complemented with liquid from the wash solvent bottle to total the programmed dispense volume Users Manual FAMOS D931R1 3 21 The User Interface lt WAIT gt lt REPEAT gt lt WASH gt Defines a pause H MM SS maximum of 9 hours 59 minutes and 59 seconds IES Note During the pause the needle will move to the 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 pL must be programmed at the desired position Enter the number of steps that must be repeated and how often they must be repeated Enter the volume for needle wash Buffer is rinsed to waste 3 8 14 The Method Menu Item lt USER PROGRAM gt If the USER PROGRAM option is enabled in the System menu the soft key lt USER PROGRAM gt is presented lt USER PROGRAM gt if enabled in System Menu Usage Menu lt EDIT gt lt INSERT gt lt DELETE gt lt ASPIRATE gt The user program offers the possibility to program all possible actions required for a sample handling sequence in separate steps IE Note The total number of
140. ubing 15 uL and 500 uL buffer and a 50 uL sample loop the maximum injection volumes are shown in TABLE 6 5 TABLE 6 5 Maximum Injection Volume Injection Mode Maximal Injection Volume Eull 50 uL sample loss equals 130 uL two loop volumes ull loo j overfill 30 uL pre flush Partial loopfill 25 uL uL pick up 2 5 uL D For Volumes Larger than 200 uL With the 2000 uL buffer tubing use the appropriate sample loop size and the appropriate syringe The syringe volume should be at least 2 x injection volume IBS Note Injection volumes larger than 500 uL are possible but the sample may DANGER contaminate the syringe Make certain that you have programmed a sufficient wash after use 6 3 6 Replacing the Main Fuses and Setting the Operation Voltage Danger Disconnect the instrument from the electrical supplies before inspecting changing the fuse To change the Fuses a Pull out the fuse holder FIGURE 6 3 item 1 A AOZL OLL AOSZ HLIM AINO 3Sn 220 240 VV RE USAO IANG BAGNI s 7 220 240 V V Orientation Marks FIGURE 6 3 The Fuse Compartment b Replace the blown fuse s by fuse s of the same rating 115 V AC 10 two 5 AT fuses slow 4 x 1 UL CSA 230 V AC 10 two 2 5 AT fuses slow 5 x 20 mm IEC127 The fuses used are UL listed and CSA certified 6 8 D931R1 User s Manual FAMOS Maintenance and Troubleshooting CAUTION c Re install the fuse holder
141. ume after the analysis time to compensate for diffusion of mobile phase from the rotor groove into the first part of the sample line during the analysis time The flush volume between injections is not programmable and is always 50 of the programmed flush volume If the total amount of sample withdrawn with the next injection from the well will exceed the total volume of the buffer tubing the buffer tubing is emptied into the wash position before the next injection The next fill sequence will then start with a full flush volume A 1 2 Using an Air Segment An air segment can be used to reduce the amount of flush volume This air segment is at the front of the flush volume It will not be injected and it will not influence the injection Use of an air segment can be enabled in the System Menu General Menu Needle Injection loop _ Buffer tubing gt A gt B eluent oS m A with air segment B without air segment FIGURE A 6 Full Loop Injection with and without Air Segment With the FAMOS Fused Silica needle the flush volume must be at least 5 0 uL for injections without air segment if a stainless steel needle is used the flush volume should be at least 15 0 uL If the samples are highly viscous it may be necessary to program larger flush volumes and reduce the syringe speed for better performance User s Manual FAMOS D931R1 A 3 Injection Principles A 2 Partial Loopfill
142. ut edge next inj e System clock e Device identifier e Date e Time e Input edge next well e Freeze input active e Reset outputs after series e Air segment e Use labeled wells e Headspace pressure e Use templates e Time base display e Use calibration wells e Key click e Error buzzer e Alarm buzzer FIGURE 3 4 The System Menu 3 8 3 The System Menu Item lt GENERAL gt eS Note The microautosampler can be fitted with different syringes tubing lengths etc to meet the specific needs of the analysis The General Menu is used to enter the system configuration it is cntical that they are entered so that various operations are performed properly e g the minimum flush volume is dependent on the needle volume Once these settings are established they are not changed frequently lt GENERAL gt This soft function key is used to access the following parameters These values are entered by the user so that the system can determine the appropriate values for certain Operations AS an example the minimum flush volume is dependent on the specific needle and needle tubing The default flush volume equals two times the volume of needle and tubing LOOP VOLUME NEEDLE TUBING SYRINGE VOLUME The maximum aspiration speed of the syringe used in injection methods is dependent on the viscosity of the samples As an alternative the syringe speed can be reduced by entering a scale factor The actual syringe Speed will be t
143. witch on headspace pressure Escape return to the System Menu lt PLATES gt E enter the Plates Menu lt 96 LOW gt define the type of plate to be used lt IN ROWS gt to define the processing order of wells Escape Escape to return to the Ready Menu For this example all other settings used will be the default value In the tables presented in this appendix the name of the soft key to be selected is indicated in angle brackets e g lt PLATES gt and the value which is indicated in square brackets e g 05 0 The bold letters indicates that you have to press a function key e g E which represents Enter c Program the method by entering the parameters indicated in TABLE C 2 TABLE C 2 Programming Method Parameters Methods to enter the Methods Menu lt INJECTION gt 01 E program injection method number 1 lt PARTIAL gt E to select partial loopfill injection method 100 E to define an analysis time of 1 minute 05 0 E to define a flush volume of 5 0 uL 1 E to define the number of injections per well 1 00 E to set the injection volume at 1 00 uL Escape Escape to return to the Ready Menu C 2 D931R1 User s Manual FAMOS Additional Programming Examples d Program the series by entering the parameters indicated in TABLE C 3 TABLE C 3 Programming Series Parameters Series to enter the Series Menu 01 E to define the Series number 01 E to define the injection method number CL E to enter lt NONE

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