Home

user manual

1. OS NNN RAN gm KN Q Loosen the needle connection nut 3 Loosen the Rheodyne Rheflex and ferrule nut 1 Carefully pull out sample needle and tubing Insert a new sample needle and tube through the needle holder 5 and tighten the nut 4 Connect the other end of the tube to port 4 of the Rheodyne injection valve using a Rheodyne nut and ferrule Do not tighten too much to prevent block of tubing Lower the cover of the Triathlon Check sample needle height default height 2 mm If necessary adjust the value in the System Menu General Menu refer to Chapter 5 Select soft function key lt WASH gt in the Ready Menu to clean the new sample needle Figure 2 3 Needle assembly 0060 122 21 User manual Triathlon 2 6 Optional Needles Needles of the following types can be installed in the Triathlon e SV needle sample needle with large inner diameter for viscous samples or in case large volumes are loaded in the loop e PEEK needle biocompatible needle e Fused silica needle needle with small inner diameter for small injection volumes e Extended needle needle for switching valve placed in the side panel of the Triathlon e Silica coated needle needle for use with liquids that would be damaging
2. 8 2 4 dde tu Sinaia 9 2 5 Needles assem bly pe NY ei ues Nessuna e dens 10 2 6 Opti lial needles vedde 11 2 7 Combination of syringe sample loop and buffer tubing 11 2 8 Tray segments and Vial types 14 2 9 EEE IET 18 2 10 Loading the sample tray 18 Chapter 3 Getting started eee eee eee eee eee eee eee 9 3 1 Menus ot the Triathlon 19 3 2 Convenient working order 20 3 9 Types of Methods and links to Series 20 3 4 Executing a Series T 21 3 5 Executing a Series in remote 21 Chapter 4 Using the Triathlon ccccccccscscscscscscscsesesesssssssscecs 23 Example 1 10 pL partial loopfill injection 23 Example 2 A 10 uL injection with UL pick up 25 Example A 1 10 dilution followed by a 10 uL partial loopfill 27 Example 4 Defining a template and adding a protection code 29 Chapter 5 Reference OT 5 1 Ready MEN M cR 31 5 2 a 33 5 3 MENGAS MENU aa 37 5 4 SEES MENU c 43 Chapter 6 I O Connections FD 6
3. Comments 90 0060 122 21 Timebase methods Method number User manual Triathlon Action Time Action Time AUX 1 1 AUX 1 ON AT TIME 1 AUX 1 OFF AT TIME 2 AUX 1 ON AT TIME 2 AUX 1 OFF AT TIME 3 AUX 1 ON AT TIME 3 AUX 1 OFF AT TIME 4 AUX 1 ON AT TIME ISS A 1 ISS A POSITION 6 1 AT TIME 4 AUX 1 OFF AT TIME 2auxzonaTtme ZAUXZOFFATTME aauxzonaTtme aauxzomrattme HAXZONATIME 4auxzomrattme 1 AUX 3 ON AT TIME 2 AUX 3 ON AT TIME 2 AUX 3 OFF AT TIME 3 AUX 3 ON AT TIME 3 AUX 3 OFF AT TIME 4 AUX 3 ON AT TIME AUX 3 ISS APOSTION2ATTME BISSBPOSTION2ATTME 41SS 8POSTIONG1 ATTME AISS BPOSTION2ATTME pssom ame zespo assvroaT sv m eSsv PORT Tssvron alssaRosmons arTme SSV 4 AUX 3 OFF AT TIME haxaomame Paxson me aauxaorarnme aauxaonattme _ saxaorATTME _ 4auxaonattme _ 0060 123 21 rme arme 3 CODE OUT arme arme sconto ae tee 91 User manual Triathlon Mix method Method number Line Action Value 00 DI Ol BY O Al COT COT COT COT COT COT COT POT POT
4. accuracy since loop volume is specified with an accuracy of 10 Minimum sample loss 230 uL 2 x loop overfill flush volume for needle for the standard 100 uL loop Partial loopfill gives maximum accuracy depends on syringe accuracy and reproducibility better than 0 5 RSD for injection volumes gt 5 uL Minimum sample loss Flush volume 30 UL 30 uL is the recommended minimum flush volume combined with an air segment smaller flush volumes can be programmed but will result in decreased performance uL Pick up means zero sample loss maximum accuracy same as partial loopfill but slightly diminished reproducibility RSD better than 1 for injection volumes gt 5 5 uL of air is injected together with the sample if an air segment is selected in the System Menu For some cases other combinations of syringe loop and or buffer are advised Injection volumes smaller than 5 pL e Partial loopfill use a 100 uL syringe for maximum reproducibility and accuracy Use 20 uL sample loop to avoid loss of accuracy due to expansion of the loop content when switching from inject to load position prior to sample loading Specially when working with high pressure 200 bar this loss may be 0 1 0 5 uL for a 100 uL loop Note that the minimum sample loss in partial loopfill mode is 30 uL recommended minimum flush volume for the first injection and an additional 15 always half the programmed flush volume for additional
5. rir eo 227 JV bM M ZU aepo VG VER Figure 2 6 Conical vials with support 5 06 06 ml 06 ml PE NN Dos Figure 2 7 Plastic vials Chromacol inserts 02 MTV 02 MTVWG 03 MTV can be used in combination with the appropriate vial and support sleeve or spring V See eZ 4 5V 02 MIV MIs 0 2 ml Figure 2 8 Standard vials with insert User manual Triathlon Eppendorf micro centrifuge tubes for standard tray if F 15 ml i Figure 2 9 Eppendorf micro centrufuge tubes with support sleeve Eppendorf micro centrifuge tubes can be used with a support sleeve However the prepuncuring needle may not be able to pierce the caps of some brands of tubes because of the depth of the caps This may result in damage to the sample needle Chromacol vials for LSV tray outer vial diameter 15 mm A L
6. The front of the Triathlon contains the following elements Figure 1 1 Front view of the Triathlon E a SER 2 10 11 12 Cover in open position Buffer tubing Injection valve Rheodyne Tubing holder Needle unit Tray with segments Drain wash position Condensed water and leakage Wash position Wash solvent bottle Syringe Syringe waste tubing 0060 122 21 User manual Triathlon 1 3 Back view of the Triathlon The back of the Triathlon contains the following elements VQ UUO Figure 1 2 Back of the Triathlon 1 SparkLink communication interface connector 2 CE mark 3 connectors 4 Mains switch 5 Mains input 6 Fuses and voltage selector 7 UL label 8 Type label 9 Fan if tray cooling is installed 10 Ventilation holes 0060 123 21 3 User manual Triathlon 1 4 Keypad and display of the Triathlon The keypad of the Triathlon contains the following elements Soft function keys Cursor keys Numeric keys Function keys Figure 1 3 Keypad and display Soft function keys the label assigned to these keys depends on the menu that is active The label of each key is shown in the bottom line of the display Cursor keys can be used to move to different field in the display to move to different field in menu or to make displayed value higher or lower Numeric keys e 0109 ente
7. Figure 6 2 TTL output 48 0060 122 21 User manual Triathlon 6 3 inputs The following connector is an active high or active low TTL input it can be defined in the System Menu The NEXT INJECTION INPUT and the NEXT VIAL INPUT can be used when the Triathlon works in REMOTE CONTROL The FREEZE INPUT and STOP input can be used to control the Triathlon 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 connection diagram is shown in figure 6 3 page 46 Table 6 9 Connector P6 INPUTS TTL NEXT INJECTION INPUT Signal ground NEXT VIAL INPUT Signal ground FREEZE INPUT Signal ground STOP Signal ground INPUT 1 Signal ground INPUT 2 Signal ground INPUT 3 Signal ground INPUT 4 Next injection input This input will start the next injection sequence when the Triathlon is started in remote control When the injection sequence is finished the Triathlon will wait for the next input From the Ready Menu a NEXT INJECTION INPUT will start the last programmed series In this case the Triathlon will not wait for the NEXT INJECTION INPUT before continuing with the next injection The Triathlon will execute the complete RUN as if it was started with the Start Stop key Next vial input With this input the Triathlon will perform the next injection from the next vial even if not all injections from that vial in the programmed injection
8. Missing destination vial Not enough transport liquid available due to missing transport vials or segments Flexprint of the sample needle is not connected Invalid configuration of the Triathlon PCB missing Current limit of the external I O exceeded Error occurred during initialization the Triathlon cannot start 0060 122 21 User manual Triathlon Appendix D Options and accessories number 0900 000 0900 830 0900 831 0900 832 0900 836 0900 840 0900 872 0900 978 0880 914 0880 915 The following configuration options are available for the Triathlon description Triathlon Autosampler Integrated Stream Switching option two six way valves Integrated Stream Switch option one six way valve Triathlon SSV Solvent Selector Valve SSV a six way solvent selector valve Triathlon Integrated Stream Switch two six way biocompatible Rheodyne 9000 series valves Tray cooling Peltier type cooling elements Biocompatible option Rheodyne 9000 series injection valve Peek needle and Peek loop Micro option Triathlon Mistral HPLC column thermostat to be used in combination with the Triathlon autosampler Mistral Peltier cooled HPLC column thermostat to be used in combination with the Triathlon autosampler Trays and tray segments 0900 313 0900 315 0900 314 0900 316 0900 317 0900 351 0900 352 0900 353 0900 354 0900 355 0900 356 0900 357 0900 358 0900 361 0900 362 0900 363 0900 364 0900 3
9. calibration Triathlon 63 68 calibration vials 36 42 carry over test 65 67 CE mark 3 CL 4 19 lt CLOCK gt 37 lt CODE gt 40 42 lt COMM gt 37 communication interface 3 lt COMPRES gt 41 condensed water and leakage 2 8 32 contact closures outputs 45 46 lt COOL gt 32 cooling 3 8 32 calibration 63 lt COPY gt 31 lt COUNT gt 32 cover 2 create template 29 cursor keys 4 19 D lt DATE TIME gt 32 lt DEFAULT ALL gt 32 define series 24 26 29 lt DELETE gt 35 38 lt DESTINATION gt 28 0060 123 21 User manual Triathlon device identifier 37 lt DISPENSE gt 38 40 display 4 drain wash position 2 8 4 19 lt EDIT gt 38 40 lt END gt 40 lt ERASE gt 31 error beep 34 error codes 57 58 Escape 5 19 lt EVENTS gt 32 examples 23 30 lt EXIT gt 33 experienced users extended needle 11 F factory installed items 52 fan first time users first transport vial 36 fluid connections 1 flush volume 12 13 39 flushed loop 1 11 13 75 77 freeze input 49 freeze input active 36 fused silica needle 11 fuses 3 54 G GENERAL 33 34 Getting started 19 21 H headspace pressure 10 34 HEX 36 Help 5 HIGH 41 Hold Cont 4 HOME 41 HPLC connections 7 95 User manual Triathlon I initialization 51 lt INJECT gt 41 INJECTION 39injection principle 1 75 80 injection method 20 number 37 injection valve see Rheodyne valve injection volum
10. 32 MARKERS 42 METHODS 38 42 MIX 38 NEEDLE 41 lt gt 33 lt PRIME gt 32 lt PROG OUT gt 41 lt REMOTE gt 21 lt REPEAT gt 38 lt SERIAL gt 33 lt SERVICE gt 33 lt SSV gt 32 42 lt START gt 21 24 lt SYR gt 41 lt SYR END 8 9 31 lt SYR HOME gt 8 9 31 SYH VALVE 41 TEMPLATE 30 37 lt TIMEBASE gt 39 TRAY 36 lt UNLOAD gt 41 USAGE 35 36 USER PROGRAM 40 lt UTILS gt 31 VALVES 39 41 WAIT 38 41 WAIT IN 41 WASH 9 31 38 39 41 WASTE 28
11. connecting passages in the rotor seal Rotation of the valve 60 switches the valve from one position to another Specifications Maximum temperature is 50 The valve is set to hold 345 bar 5000 psi The wetted surfaces are PEEK alumina ceramic and an inert polymer Use only plastic ferrules in the stator ports Metal ferrules can cause irreparable damage to the plastic stator 70 0060 122 21 Maintenance With normal use the valve will give many tens of thousands of cycles without trouble The main cause of early failure which is seen as a leak in the valve is abrasive particles in the sample and or mobile phase which can scratch the rotor seal Following is the procedure for changing the rotor seal stator face assembly and isolation seal Refer to figure F 4 and proceed 10 Remove the three stator screws with a 9 64 inch hex key Remove the stator stator face assembly and stator ring from the valve body Pull the rotor seal off the pins Remove the isolation seal Mount the new isolation seal with the spring side facing away from the rotor seal Mount the new rotor seal The three pins on the shaft assembly fit into the mating holes in the rotor seal only one way Mount the seal with the grooves facing the stator Replace the stator ring so that the body locating pin in the stator ring enters the mating hole in the body Put the new stator face assembly on the stator The three pins on the a
12. horizontal The sample needle arm needed too many or too few steps to reach destination Sample needle arm not in vertical home position while moving horizontally The sample needle is at an invalid horizontal position The sample needle detects too many code gaps while moving to the next position The samlpe tray is not at a valid position while moving down the needles Stop sensor is not in tray gap Vial sensor sticks The sample needle spindle does not rotate correctly The sample needle did not reach or leave home position The sample needle is not at home position 57 User manual Triathlon Tray ERROR 51 ERROR 52 ERROR 53 Vials ERROR 60 ERROR 61 ERROR 62 ERROR 63 ERROR 64 ERROR 65 ERROR 66 ERROR 67 ERROR 68 ERROR 69 Electronics ERROR 71 ERROR 72 ERROR 73 ERROR 75 58 Incorrect tray rotation No segment found The sample needle arm is not in the home position while moving the tray Missing vial Only available when Skip Missing Vial is set to NO in the System Settings Missing segment Only available when Skip Missing Vial is set to NO in the System Settings and during the execution of a Mix Method Missing transport vial Missing transport segment Only available when Skip Missing Vial is set to NO in the System Settings and during the execution of a Mix Method Missing vial for reagent Missing vial for reagent B Missing vial for reagent C Missing vial for reagent D
13. lt INJECTION gt Use this key to program a method that defines injection methods max 24 for a run of the types full loop partial loopfill or uL pick up Enter a number for the injection method you are going to program If the selected method is locked because of changes in the settings System Menu 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 Use the soft function keys to select an injection method then enter values for e analysis time the time between switching the injection valve to inject and the start of processing the next sample e flush volume the amount of sample taken from a vial before the loop is filled with sample Default value 30 uL combined with an air segment Flush volumes of less than twice the volume of the needle and tubing will result in decreased performance e number of injections per vial maximum value is 9 e injection volume can be entered for each injection per vial The maximum programmable injection volumes are partial loopfill 5096 of the programmed loop volume HL pick up injection volume loop volume 3 x needle volume 2 flushed loop not programmable is equal to the loop volume but needs more sample to fill the loop 3 x loop volume for loop volumes 100 uL 2 x loop volume for loop volumes gt 100 uL 499 uL 1 5 x loop volume for loop volumes
14. self test initialization procedure are executed After completion of this procedure the following appears on the display 10 26 TUESDAY DECEMBER 16 97 MENU READY lt ADVANCE gt lt WASH gt lt SYR END lt UTILS gt 0060 123 21 51 User manual Triathlon Factory installed items The Triathlon is factory installed with e fuses in power switch 115 V AC 10 two 5 AT fuses slow 2 x 1 UL CSA or 230 V AC 10 two 2 5 AT fuses slow 5 x 20 mm IEC127 100 UL loop 250 uL syringe 500 uL buffer tubing stainless steel 15 uL sample needle The fuses used are UL listed and CSA certified Standard sample needle and tubing SS tubing 135 mm x 0 65 mm O D x 0 25 mm 1 0 Tefzel tubing 140 mm x 46 O D x 0 25 mm I D total volume 15 uL Buffer tubing from high pressure valve to syringe valve PTFE tubing 640 mm x 46 O D x 1 0 mm I D volume 500 uL Tubing syringe valve to wash solvent bottle PTFE tubing 300 mm x 6 O D x 1 0 mm I D Tubing syringe valve to waste PTFE tubing 400 mm x O D x 1 6 mm I D For installation of HPLC connections waste tubing wash solvent syringe sample needle trays etc refer to Chapter 2 of this manual 52 0060 122 21 User manual Triathlon Appendix B Specifications General Sound pressure level Working temperature Storage temperature Humidity Sample viscosity Safety Sample capacity Tray segment types Vial diameter
15. 714 0900 715 0900 718 0900 721 0900 811 0900 812 2140 160 3796 004 3796 005 3796 035 3796 009 3796 010 3796 019 description Tray cooling cover Needle wash insert vial Needle body User manual Long Bushing Rheodyne pck 10 Ferrule Rheodyne pck 10 Rheflex ferrule pck 10 Rheflex nut pck 10 Flanged tube fitting 1 8 pck 5 Flanged tube fitting 1 16 pck 5 Plunger replacement tip 100 uL pck 10 Plunger replacement tip 500 uL pck 10 Plunger replacement tip 1 mL pck 10 Biocompatible injection valve assembly Injection valve assembly Sample needle including tubing ferrules and nuts oerum sample needle including tubing ferrules and nuts Peek sample needle including tubing ferrules and nuts Sample needle extra long including tubing ferrules and nuts Serum sample needle extra long including tubing ferrules and nuts Peek sample needle extra long including tubing ferrules and nuts Fused silica needle including tubing ferrules and nuts Fused silica needle extra long including tubing ferrules and nuts ooftware Triathlon Buffer tubing 500 uL Buffer tubing 2000 uL oyringe waste tubing oyringe wash tubing Tubing Solvent Selector valve Syringe valve Buffer tubing 15 mL Air prepuncturing needle Tefzel buffer tubing 2000 uL Inject marker cable Next injection input cable Luerlock connection syringe valve Rheodyne 7010 rotor seal Rheodyne 7010 stator Rheodyne loop 5 uL Rheodyne loop 10 uL Rheodyne
16. Escape to return to the Ready Menu To run the series Start Stop to start the Triathlon 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 Triathlon now performs the same actions as in Example 2 except that analysis is performed on two vials 01 and 01 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 30 0060 122 21 User manual Triathlon Chapter 5 Reference This chapter describes all possibilities offered by the Triathlon software in the order in which they appear in the screen 5 1 Ready Menu The Ready Menu contains the following soft function keys lt ADVANCE gt Use this key to rotate the tray of the Triathlon You can now fill the tray with segments by placing the segment at a free position in the front half of the tray lt WASH gt Use this key to start a standard wash procedure All tubing connected to the syringe valve will be filled and rinsed with wash solvent lt SYR END gt Use this key to move the syringe to end position if you wish to replace the syringe needle or to simplify filling of wash solvent tubing syringe volume of wash solvent is aspirated from the wash solvent bottle and the wash solvent tube is filled Select soft function key lt SYR HOME gt to dispense the syringe contents to syringe waste and
17. POT PT PT D D D DI D gt O OF CO NI DJ BY O POT O CO CO Ny DJ O17 BY O POT O Of CO Ny DJ Cy BY Coy D O Comments 92 Position Speed Height Line 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 Action Value Position Speed Height 0060 122 21 User program User manual Triathlon Method number Line Action Value 00 DI Ol BY O Al COT COT COT COT COT COT COT POT POT PT PT PT NINININI D oe gt O OF 00 OJ Of BI O POT CO CO CO OD O1 BY Cy D CO CO NJ OI OT BY OI PO Comments 0060 123 21 Position Speed Height Line 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 Action Value Position Speed Height 93 User manual Triathlon 94 0060 122 21 Index UL pick up 1 11 13 78 80 example 23 24 accessories 59 61 accuracy 12 13 air outlet nut 10 11 air segment 34 73 74 76 77 alarm buzzer 34 analysis time 39 lt ASPIRATE gt 38 40 lt AUTOMATIC gt 32 lt AUX gt 39 41 B BCD 36 buffer tubing 1 2 11 13 buttons see keys
18. _ VEN U E 4 04 OFA 5 90 Figure 2 10 Vials and inserts 16 0060 122 21 User manual Triathlon Chromacol vials for super LSV tray outer vial diameter 22 mm 6 0 ml 10 0 ml Dey 10 CV Figure 2 11 Vials for super LSV tray Chromacol vials for micro tray outer vial diameter 7mm 0 8 ml 1 0 ml 0 7 ml 0 8 ml 0 6 ml 0 5 ml NSS MX Xsan pet Nn U DELEN VER SERVED WY Figure 2 12 Vials support User manual Triathlon 2 9 Vial handling It is best to fill vials with a narrow end pipette to allow air to escape when filling the vial Do not fill vials to the edge to avoid that sample liquid will be forced into the air needle risking cross contamination of samples and fouling of the needles If you want to use headspace pressure for a sample handling routine it is important to make sure that the seal is airtight An airtight seal is also important to prevent air bubbles from forming and to prevent evaporation of volatile samples Check the seal after crimping if the cap can be turned easily the seal is not airtight and you will have to adjust the handcrimper 2 10 Loading the sample tray The tray segments can be placed in any position in the Triathlon tray The tray segment types used must be defined in
19. a programmed method to a range of vials in the Series Menu 4 Execute the series Please note that it is allowed to use a different order however you should keep in mind that settings entered in the System Menu determine which possibilities appear in the Methods and Series menus Refer to chapter 5 for more information on specific items in menus and the way they influence the other menus 3 3 Types of Methods and links to Series The Triathlon offers the following types of methods for different parts of the sample handling routine 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 with 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 Triathlon in separate steps Each programmed method is assigned a number The Triathlon offers the possibility to store a combination of defined methods in a template A template is also identified by a number Methods must be linked to series before they can be used The following possibilities are offered by the Triathlon e You can assign an individual method
20. has positions 1 6 and 1 2 see figure 5 2 to program a pause max 9 hours 59 minutes 59 seconds During the pause the needle will move to home position if the previous step is an aspirate or dispense action If you want the needle to stay in the same position an aspirate or dispense step of 0 uL must be programmed at the desired position to activate the compressor to put air pressure on a sample The compressor will stay active until it is switched off in a next programmed step The compressor will be automatically switched off at the end of the needle wash routine if a needle wash is used to control the four standard auxiliaries contact closures Refer to Chapter 6 to program a pause in which the Triathlon waits for one of the four inputs to become HIGH or LOW before continuing with the next step Refer to Chapter 6 to define two programmable outputs contact closures These are similar to the auxiliaries but only available in the user program Refer to Chapter 6 41 User manual Triathlon lt CODE gt to program the output to the connector P3 TIMED OUTPUTS This is a HEX output in the range 0 to 15 Refer to Chapter 6 lt MARKERS gt the markers normally generated in the Triathlon are not active in the user program but can be programmed in this screen refer to Chapter 6 Select marker and status inject vial labeled lt SSV gt option to define the Solvent Selection Valve SSV port pos
21. method have been executed Freeze input The Triathlon will freeze the analysis time for the time this input is active If the FREEZE INPUT is activated while the analysis time is not running the Triathlon will perform all programmed pre injection sample handling mix method and loading part of the injection method But the Triathlon will wait with injecting the sample until the FREEZE INPUT is no longer active Stop I O With this input the run of the Triathlon is immediately aborted The Ready Menu appears in the display In case the Triathlon is in remote control the run of the Triathlon is immediately aborted but the Triathlon remains in remote control and cannot be restarted with a NEXT INJECTION INPUT 0060 123 21 49 User manual Triathlon INPUT 1 4 Programmable input can be used in the user program Active high Active low Autosampler Autosampler VCC VCC A xX a z GND GND Figure 6 3 TTL input 6 4 MultiLink connector The MultiLink connector is used to make the Triathlon communicate with the SparkLink software Set the dipswitch S1 and S2 as follows to let the communication work probably pA Ne og Q 80 2 Figure 6 4 MultiLink board to next device to PC or previous device Connections for 9 pin connector pin 2 TD Transmitted data to th
22. or after emptying of the buffer tubing the syringe dispenser aspirates transport 3 TED am gt liquid from the transport vial to fill the sample line with Z transport liquid and remove wash solvent Transport liquid 0060 122 21 gt Co User manual Triathlon The needle moves from the transport vial to the sample C vial The injection valve is switched to the LOAD position lt Syringe 22243 LOAD Column gt Needle Sample Transport liquid a The programmed injection volume is aspirated from the sample vial I Syringe Coum gt Sample Transport liquid The sample needle moves back to the transport vial The sample is quantitatively transported into the loop with transport liquid mobile phase from the transport vial lt a Syringe Transport liquid The injection valve is switched to INJECT The sample loop is now part of the HPLC mobile phase flow path sample is transported to the column The analysis time lt Syringe AZ gt Starts to run Transport liquid 0060 123 21 79 User manual Triathlon The next sequence will skip the first withdrawal of transport solvent unless a wash routine is performed or the Triathlon has emptied the buffer tubing to waste In those cases the sequence is completely repeated Air segment If an air segment has been programmed it appears at the front of the
23. rinsed Select the appropriate syringe in the System Menu and install the correct buffer tubing refer to section 2 7 Figure 2 2 Replacing the syringe 0060 123 21 9 User manual Triathlon 2 5 Needle assembly The needle used for sampling consists of two parts prepuncturing needle a hollow needle used for puncturing of the septum capmat or sealer also used to put headspace pressure on the sample approximately 0 5 bar sample needle placed inside the hollow prepuncturing needle used for the actual transport of sample Different types of needles can be used here refer to section 2 6 and options mentioned in Appendix D If a needle with deviating diameter is used a different air outlet nut see figure 2 3 number 6 must be used that matches the injection needle Most commercially available sealers or capmats cannot be used in combination The following elements can be identified in the needle assembly Numbers 1 2 3 4 and 6 constitute the sample needle Execute the following steps to replace a needle 1 2 9 4 EE LS IE I with headspace pressure You are advised to switch off headspace pressure in those cases System Menu General Menu Rheodyne Rheflex nut and ferrule Needle tubing Needle connection nut Standard sample needle Needle holder z Air outlet nut Prepuncturing needle Sensor detects tray height missing tray and missing vials
24. to a stainless steel needle Refer to Appendix D for an overview of options available for the Triathlon Execute the following steps to install any of these needles 1 Remove needle as described in section 2 5 2 Remove the standard air outlet nut figure 2 3 number 6 and replace it by the nut supplied with the optional sample needle 3 Install optional sample needle as described in section 2 5 4 Adjust settings in System Menu General Menu to the volume of the new needle tubing see section 5 2 2 7 Combination of syringe sample loop and buffer tubing The 250 uL syringe is the standard syringe combined with the standard 500 uL buffer and the standard 100 uL sample loop the following injection volume range is available for the various injection modes Flushed loop 100 uL Partial loopfill 1 50 ul UL pick up 1 27 ul The maximum injection volumes are calculated with the following formulas Flushed loop injection volume loop volume Partial loopfill injection volume 50 of loop volume UL pick up max injection volume loop volume 3 x needle tubing volume 2 0060 123 21 14 User manual Triathlon Five sizes of syringes can be used in the Triathlon 100 250 500 1000 uL and 10 mL The 10 mL syringe can only be used in the User Program must be enabled in System Menu see section 5 3 and not for the standard injection modes Flushed loop gives maximum reproducibility RSD lt 0 3 but not maximum
25. to move the syringe to standard operating position again lt UTILS gt Use this key to go to the Utilities Menu If use of a method protection code is enabled in the System Menu the code must be entered to access the Utilities Menu The menu offers the following possibilities lt COPY gt to copy a method Enter the type mix injection timebase wash and the number of the method to be copied Then enter a number to define the destination method Any existing method stored under that number will be overwritten lt ERASE gt 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 Note that it is not possible to erase the user program if the protection code for the user program is enabled in the System Menu User manual Triathlon lt LOG gt the Triathlon 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 message appears after every 50 000 syringe actions and after every 200 000 syringe valve actions Lifetime of syringe valve maybe exceeded Check for possible leakage Syringe if you do not replace the syringe at this moment and tell the system not to display this message again the message will not be displayed again until 50 000 more syrin
26. up Example 1 A 10 pL partial loopfill injection After the Triathlon has been switched on an initialization procedure is executed In this example it is assumed that a loop of 100 uL needle tubing of 15 uL a syringe of 250 uL and a type A tray has been installed Wait until the Ready Menu appears on the screen then execute the following steps For system settings System to enter the System Menu lt GENERAL gt E to enter the General Menu 0100 E to define the volume of the installed loop 015 E to define the volume of the needle tubing lt 250 gt E to define the volume of the syringe lt NORMAL gt E to set syringe speed to normal 02 E to set sample needle height to 2 mm lt YES gt to enable use of air segment lt gt to switch headspace pressure Escape Escape to return to the Ready Menu For this example all other settings used will be default 0060 123 21 23 User manual Triathlon To program a method 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 030 E to define a flush volume of 30 UL 1 E to define the number of injections per vial 10 E to set the injection volume at 10 ul Escape Escape to return to the Ready Menu To define the series to enter the Series Menu to define the Series number to define the injection method n
27. 1 Contact closures outputs 45 6 2 I Neve me 46 6 3 crc 49 6 4 COMET ene 49 0060 123 21 User manual Triathlon Appendix A Installation 51 Appendix B Specifications ccccccsccscccscsscscscscescscccscssescscecess 53 Appendix Error codes 57 Appendix D Options and accessories 59 Appendix E Calibration and performance 63 Appendix Rheodyne injection valve 69 Appendix Programming options eee ee ee ee ee ee ee eere eee nene 73 Appendix Injection principles eee ee ee ee ee ee eene ue 75 Appendix I Trouble shooting e eee ee ee ee ee ee ee eee eu 81 Appendix J Logbooks eee v Ie Y ve Va VE VE EA I 85 naek 95 vi 0060 123 21 User manual Triathlon Chapter 1 Introduction The Triathlon is an autosampler that offers a wide variety of possibilities for HPLC It incorporates among other things the following features e suitable for use with various types of vials by various manufacturers e capable of executing flushed loop injections partial loopfill injections and uL pickup see section 1 1 below e accomodates use of various types of syringes volumes of 100 250 500 1000 LL or 10 mL e fully co
28. 1 of the Rheodyne injection valve e HPLC column to port 6 of the Rheodyne injection valve The instrument has been flushed with isopropanol before dispatch 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 It is very important that the contents of the sample loop are injected in back flush onto the column therefore do not exchange column and pump connections at the injection valve 0060 123 21 7 User manual Triathlon 2 2 Waste tubing Make the following connections Syringe waste put the end of the syringe waste tube see figure 1 1 number 12 ina bottle placed next to the Triathlon Drain wash position connect the hose to the drain wash connector of the Triathlon see figure 1 1 number 7 place the other end of the hose in a bottle placed on the floor Through this drain all liquid dispensed to waste at the back of the tray is removed Condensed water and leakage connect the hose to the drain port of the Triathlon see figure 1 1 number 8 Place the other end of the hose in a waste container on the floor Through this hose all leakage solvents and condensed water if Peltier cooling is installed are drained Make sure that the flow path of the hoses is not obstructed in any way 2 3 Wash solvent The Triathlon has a 250 mL wash solvent bottle Execute the following steps to install t
29. 23 21 75 User manual Triathlon For flushed loop injections the sample loop is quantitatively filled by transporting two or more times the loop volume through the loop depending on the volume of lt Syringe j Coum gt the loop Sample The injection valve switches in INJECT position The sample loop is now part of the HPLC mobile phase flow path sample is transported to the column The analysis o time starts gt lt CB mos Sample If one injection is to be done from each vial or if wash routine has to be performed after every injection the needle withdraws from the vial directly after the injection and if programmed directly performs wash After the analysis time new sequence Is started If more than one injection is done from the same vial without wash routine the Triathlon withdraws a flush volume 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 vial 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 Air segment An air segment can be used to reduce the amou
30. 65 0900 366 0900 367 0900 368 0060 123 21 Tray set for cool option 8 segments 1 8 mL vial 12 positions 12 mm O D Tray set standard 8 segments 1 8 mL vial 12 positions 12 mm O D Tray segment 0 5 mL vial 20 positions 7 mm O D Tray segment 4 mL vial 9 positions 15 mm O D Tray segment 10 mL vial 4 positions 22 mm O D Tray segment standard tray number 1 Tray segment standard tray number 2 Tray segment standard tray number 3 Tray segment standard tray number 4 Tray segment standard tray number 5 Tray segment standard tray number 6 Tray segment standard tray number 7 Tray segment standard tray number 8 Tray segment tray with cooling option number 1 Tray segment tray with cooling option number 2 Tray segment tray with cooling option number 3 Tray segment tray with cooling option number 4 Tray segment tray with cooling option number 5 Tray segment tray with cooling option number 6 Tray segment tray with cooling option number 7 Tray segment tray with cooling option number 8 59 User manual Triathlon 60 number description The following accessories are available for the Triathlon number 0043 344 0043 364 0043 473 0060 123 0700 002 0700 003 0700 008 0700 009 0700 010 0700 011 0700 012 0700 018 0700 019 0900 300 0900 301 0900 303 0900 304 0900 311 0900 321 0900 322 0900 323 0900 325 0900 326 0900 700 0900 710 0900 711 0900 712 0900 713 0900
31. Dimensions Vial Loop volume Puncturing accuracy needle Pre puncturing septa caps Vial amp height detection Switching time injection valve Headspace pressure Wash solvent Dispenser syringe Wetted parts LeAq lt 70 dB 10 35 25 60 C 20 80 RH 0 1 5cP according to IEC 1010 96 vials of 1 5 mL STD 12 vials per segment 72 vials of 4 0 mL LSV 9 vials per segment 160 microvials of 0 5 mL 20 vials per segment 32 vials of 10 0 mL 4 vials per segment 4 types of tray segments see figure 2 4 Type A Standard tray segment Type B LSV tray segment Type C Super LSV tray segment Type D Micro vial tray segment 12 mm Standard tray 15 mm LSV tray 22 mm Super LSV and 7 mm Micro tray Maximum height 47 mm including cap minimum height 32 mm including cap 5 1000 uL 0 6 mm With air needle dual needle action Missing vial amp height detection by vial sensor Electrically lt 100 msec Built in compressor 250 mL internal wash solvent bottle 100 ul 250 uL standard 500 uL 1000 uL or 10 mL 55316 PTFE TEFZEL VESPEL Glass Teflon Optional PEEK Fused silica Analytical performance Capped and sealed vials e flushed loop injections e partial loopfill injections e uL pick up injections Memory effect 0060 123 21 Reproducibility RSD lt 0 3 96 RSD lt 0 5 95 injection volumes gt 5 UL with headspace pressure on the vial and 30 uL pre flush with air segment R
32. E gt 39 timebase method 20 enable disable 35 number 37 time display 34 transport vials 7 tray segments 2 14 trouble shooting 81 84 TTL inputs 49 50 TTL outputs 49 50 tubing 52 tubing holder 2 type label 3 typographical conventions i U UNLOAD 41 USAGE 35 36 USER PROGRAM 40 user program 12 13 20 21 33 37 enable disable 35 user program instead of methods 37 using the Triathlon 23 30 lt UTILS gt 31 97 User manual Triathlon v VALVES 39 41 ventilation holes 3 vials 14 voltage 51 voltage selector 3 W lt WAIT gt 38 41 lt WAIT IN gt 41 lt WASH gt 8 9 31 38 39 41 wash method 20 number 38 wash position 2 wash solvent 2 8 9 lt WASTE gt 28 waste tubing 8 vial marker pulse length 36 37 vial processing method 36 vials see trays working order 20 98 0060 122 21 Index of soft function keys lt ADVANCE gt 31 lt ASPIRATE gt 38 40 lt gt 32 lt AUX gt 39 41 CLOCK 37 CODE 40 42 COMM 37 lt COMPRES gt 41 COOL 32 COPY 31 COUNT 32 lt DATE TIME gt 32 DEFAULT ALL 32 DELETE 38 40 DESTINATION 28 DISPENSE 38 40 EDIT 38 40 END 40 lt ERASE gt 31 EVENTS 32 EXIT 33 GENERAL 33 34 HIGH 41 lt HOME gt 41 lt INJECT gt 41 INJECTION 39 lt INSERT gt 38 40 O 36 LOAD 41 LOG 32 LOW 41 0060 122 20 User manual Triathlon MANUAL
33. NK PROTOCOL Options Sample cooling Integrated stream switching ISS Solvent selection valve SVV Biocompatible option Biocompatible ISS valve Micro option Optional trays Column oven 0060 123 21 User manual Triathlon Next injection input Next vial input Freeze input Stop I O 4 Programmable inputs RS232C Microsoft Windows 95 Windows NT Yes Built in Peltier cooling processing unit Programmable Range 4 C 40 C Accuracy 2 C Cooling capacity Ambient 20 C measured on cooling tray Note Ring temperature at relative humidity of 80 and ambient temperature of 31 C decreasing to 50 at 40 C One or two optional high pressure 6 port valves 6 way integrated solvent selection valve Rheodyne 9000 series biocompatible valve with PEEK sample needle and PEEK loop One or two optional Rheodyne 9000 series biocompatible high pressure 6 port valves Triathlon for small injection volumes down to 0 1 ul equipped with VALCO valve fused silica needle and special software LSV tray 9 vials of 4 0 mL per segment Micro tray 20 micro vials per segment Super LSV tray 4 vials of 10 mL per segment Mistral column oven combined with the Triathlon The injection valve and optional high pressure valve fit into the column oven 55 User manual Triathlon 56 0060 122 21 User manual Triathlon Appendix C Error Codes The Triathlon will display an error message if the user tries to enter invalid
34. SD lt 1 0 injection volumes gt 5 pL with headspace pressure on the vial 0 01 96 with programmable needle wash o3 User manual Triathlon Programming Injection methods Injection volume Injections per vial Analysis time Needle wash Priority sample Series Physical Dimensions W x D x H Weight Electrical Power requirements Fuses Internal fuse Communication OUTPUTS 54 Flushed loop injections Partial loopfill injections UL pick up injections 1 uL 1 mL full loop depending on system settings 1 500 LL with 1 UL increments for partial loopfill 1 uL max volume with 1 uL increments for uL pick up max volume loop volume 3 x needle volume 2 Max 9 volumes are programmable for each injection Max 9 hrs 59 min 59 sec Programmable between injections vials or series Freely programmable Freely programmable 24 series max 280 mm x 540 mm x 440 mm 22 kg 30 kg if cooling option is installed 115 10 96 50 Hz 60 Hz 250 VA 230 10 96 50 Hz 60 Hz 250 VA For 115 VAC two 5 0 AT fuses 4 x Ya UL CSA For 230 two 2 5 AT fuses 5 x 20 mm IEC 127 All fuses UL listed and CSA certified One 6 3 AT fuse 5 x 20 mm IEC 127 Inject marker Vial marker Labeled vial marker Stop I O BCD output 4 Auxiliary outputs 2 Programmable outputs Alarm output 4 Bit timebase 0060 122 21 INPUTS SERIAL COMMUNICATION PORT PC CONTROL SPARKLI
35. SERT gt to aspirate an air segment of 50 UL lt ASPIRATE gt 200 to define mix method step number 11 DESTINATION gt 3 to aspirate 200 pL lt INSERT gt from the destination vial lt DISPENSE gt 200 gt 9 E to define mix method step number 12 lt INSERT gt to dispense 200 uL to the destination vial lt REPEAT 3 to define mix method step number 13 to repeat the last 2 steps three times Escape Escape to return to the Ready Menu 28 0060 122 21 User manual Triathlon To define the series 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 method number 3 CLE to select lt NONE gt for wash method 1 to define location of first sample vial 1 to define location of last sample vial 2 E to define location of first destination vial 3 E to define position of Reagent A Escape to return to the Ready Menu To run the series Put sample in position A 01 position A 02 is used as empty destination vial Place a filled reagent vial in position A 03 Make sure the reagent vial is filled correctly before starting a new series Start Stop To start the Triathlon 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 Triathlon will now start searching for the Reagent vial and transport 180 UL to the destination vial twice then 40 uL of sa
36. Triathion user manual Version 2 1 April 2000 0060 123 21 P O BOX 388 7800 AJ EMMEN THE NETHERLANDS TEL 31 591 631700 FAX 31 591 630065 E MAIL SALES SPARK NL C DECLARATION OF CONFORMITY We Spark Holland BV Pieter de Keyserstraat 8 NL 7825 VE Emmen Declare that the product Autosampler Triathlon type 900 is in conformity with the following documents e 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 EN 61010 1 Safety requirements for laboratory equipment Class I Installation cat 11 Pollution degree 11 Spark Holland will not accept any liability for damages directly or indirectly caused by connecting this instrument to devices which do not meet relevant safety standards WARNING e EEC directives 89 336 and 92 31 EMC requirements applied with the following standards EN 50081 1 Generic emission standard EN 50082 1 Generic immunity standard EN 61000 3 2 Harmonic current emissions Use shielded cables and connectors for all remote connections Emmen October 1998 Rein van den Berg QA manager User manual Triathlon About this manual This manual has been written for laboratory technicians who use the Triathlon for execution of analytical runs It is assumed that the user of this manual has basic knowlegde of how to use menu driven software and that she he is fa
37. Triathlon from all power sources before removing protective panels Always replace blown fuses with fuses of the size and rating indicated on the fuse panel and holder Refer to Appendix B of this manual for more information on fuses Replace or repair faulty insulation on power cords Check that the actual power voltage is the same as the voltage for which the Triathlon is wired Make sure power cords are connected to correct voltage sources The Triathlon must only be used with appliances and power sources with proper protective grounding Q Perform periodic leak checks on supply lines 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 Using the Triathlon in other ways than indicated in the instructions given in this manual may cause unsafe conditions 0060 123 21 lil User manual Triathlon iv 0060 123 21 User manual Triathlon Table of contents Chapter 1 INIT OGUCTION 1 1 1 1 2 PLONE VIEW OF TNE TT m 2 1 3 Back view of the Triathlon VV evden 3 1 4 Keypad and display of the Triathlon 4 Chapter 2 Preparations for use ccccccccccscscscssesescscscscscscesesesecs 2 1 FPG GCONMECIONG saa 7 2 2 Waste 1UDING AT 8 2 3 Wasi
38. alibration vials These parameters depend on the contents of the methods used and or the System Settings of the Destination vials Triathlon Reagent vials Labeled vials Note IStariStop Optional Soft function keys e Last series lt START gt lt REMOTE gt PRIORITY Only available when user program and mix Priority uela method have been disabled in the System ash metho Timebase method Settings Vial number Interrupt Not used Help Context and status sensitive help screen if available 74 0060 122 21 User manual Triathlon Appendix H Injection principle This appendix offers specialist information on the injection principles used by the Triathlon H 1 Flushed loop injections The switching sequence for a flushed loop injection is schematically shown in the following figures The initial situation the injector is in the INJECT position The sample needle has entered the vial after the air neede has prepunctured the septum Headspace pressure is applied through the outer air needle to ensure that no air or vapor bubbles are formed during sample aspiration The syringe dispenser aspirates the programmed flush volume from the sample vial to fill the sample line with sample and remove wash solvents The injection valve switches in LOAD position placing a sharp sample front at the inlet of the sample loop a lt Syringe 4 Column gt LESE EE Sample 0060 1
39. alytical performance test E 1 1 Analytical system The Triathlon is tested in an analytical system under the following conditions Pump flow 1 5 mL min eluent distilled water Detector wavelength 254 nm Sample Uracil in distilled water 50 ppm The following configuration was used PUMP 1 5 ml min UV Detector 254 nm wae ntegrator 10 mtr SS tubing 0 25 mm ID for peak modelling E 1 2 Relative standard deviation RSD The following formulas are required for calculating the RSD Peak area Peak area n Peak area Peak area OL i n 1 RSD x100 Peak area 0060 123 21 63 User manual Triathlon E 1 3 Reproducibility test The default system settings were used except for the following lt GENERAL gt Air segment NO lt TRAYS gt Location first transport vial A7 Location last transport vial A7 Sx vials are filled with sample on positions A1 A6 The transport solvent vial is filled with eluent and placed on position A7 The wash solvent bottle is filled with 80 20 Iso propanol Injection method Wash method Series Number 1 Number 1 Number 1 Type UL pick up Wash between injections First vial A 01 Analysis time 1 00 min Wash volume 300 uL Las
40. ates Time base display Use calibration vials Key click Error beep Alarm buzzer Methods Method lt INJECTION gt Injection methods lt FLUSHED gt lt PARTIAL gt lt PICK UP gt e Analysis time e Analysis time e Analysis time e Flush volume e Flush volume e Injection vial e Injection vial e Injection vial e Inj volumes Inj Volumes Series e Template e Use calibration vials yes no calibration vials calibration interval e Sample vials e Destination vials e Reagent vials e Labeled vials Start Stop e First series e Last series e lt START gt lt REMOTE gt Priority e Template e Vial number Interrupt Not used Help Context and status sensitive help screen if available 0060 123 21 lt Menu gt lt ERASE gt lt LOG gt amp Userprog lt TRAY gt 1 type number 2 type number 3 type number 4 type number 5 type number 6 type number 7 type number 8 type number number of transport vials lt WASH gt Wash volume Wash between Series Vials njections Logbook lt SERIAL gt lt SSV gt COOL lt SERVICE gt lt DEFAULT ALL gt ON OFF All methods Communication Prime solvent Service mode erased all settings protocol valve to default lt Menu gt Jo lt CLOCK gt lt COMM gt Vial output e Device identifier Inj Marker pulse Vial marker pulse Lab Vial marker pulse Input edge next inj Input ed
41. data Information on the allowed range will be displayed If something goes wrong in the physical operations of the Triathlon an error code will be displayed Press the Start Stop key twice to lift the message and try to repair the failure condition with the help of the explanation of the code concerned Injection valve and ISS unit ERROR 11 ERROR 12 ERROR 13 ERROR 14 ERROR 15 ERROR 17 ERROR 18 Injection valve is not in a valid position The injection valve did not switch within 1 5 seconds The switching time of the injection valve exceeds 500 msec ISS valve is not in a valid position The ISS valve did not switch within 1 5 seconds ISS valve B is not in a valid position ISS valve B did not switch within 1 5 seconds Syringe dispenser unit ERROR 21 ERROR 22 ERROR 23 ERROR 24 ERROR 25 Injection needle ERROR 30 ERROR 31 ERROR 32 ERROR 33 ERROR 34 ERROR 35 ERROR 36 ERROR 37 ERROR 39 ERROR 40 ERROR 41 ERROR 42 0060 123 21 The syringe valve did not switch The syringe did not reach home position in time The syringe spindle did not make the correct number of rotations The spindle does not rotate The syringe valve did not find a valid position unit The sample needle arm did not reach or leave home position vertical The sample needle arm is in an invalid horizontal position while moving down The sample needle arm did not reach or leave destination within a certain time
42. e number Vial marker pulse length type number type number Labeled vial marker pulse type number length 11 SEC type number type number Input edge next injection jfalling rising position first transport vial Input edge next vial falling rising position last transport vial Freeze input active low high Reset outputs after last series yes Ino lt CLOCK gt lt COMM gt yy mm dd and hh mm Device identifier 2 Comments 0060 123 21 87 User manual Triathlon Templates Template Injection Mix Wash Timebase User program Comments method method method method Comments 88 0060 122 21 User manual Triathlon Injection methods injection type Flush Inj Injection volumes PPP PPP pick up Bo j ee ee SEE SE AR AE AR GMA SIS Th Je l 6 l J l1 m pcc 8e l 9 l j L1 L Lt 1 d l 1 1 l1 l1 li Comments 0060 123 21 89 User manual Triathlon Wash methods Wash method Wash between Wash volume Comments number linjections vials series linjections vials series linjections vials series linjections vials series ot linjections 3 vials series
43. e 11 13 39 inject marker pulse length 36 37 INPUT 1 4 45 46 input edge next injection 36 input edge next vial 36 lt INSERT gt 38 40 installation 7 51 52 Interrupt 4 introduction 1 5 lO 36 I O connections 3 45 46 ISS 39 K key click 34 keypad 4 5 keys 4 5 L labeled vial 35 43 labeled vial marker pulse length 36 last transport vial 36 LOAD 41 LOG 32 logbooks 85 94 loop volume 33 calibration 67 LOW 41 LSV needle 11 M mains input 3 mains switch 3 lt MANUAL gt 32 MARKERS 42 MENU 5 19 Menu 5 19 Methods 5 19 20 21 METHODS 38 42 INJECTION 39 40 MIX 38 39 lt TIMEBASE gt 39 96 USER PROGRAM 40 42 WASH 40 Methods Menu 29 37 42 missing tray detection 10 missing vials detection 10 MIX 38 mix method 20 37 enable disable 35 number 37 MN 5 19 N needle 2 10 height 35 38 39 holder 10 prepuncturing 10 16 replacing 10 sample 10 standard 10 tubing 10 33 35 types 11 next injection input 49 next vial input 49 number of injections per vial 39 numeric keys 4 options 55 59 lt gt 33 partial loopfill 1 11 13 77 78 example 23 24 27 29 control 31 needle 11 Peltier cooling 8 32 56 performance Triathlon 63 68 pictograms ii tray detection 10 14 tray segments 2 14 installation 51 lt TRAY gt 36 types 14 preparations for use 7 18 lt PRIME gt 32 Priority 4 priority samp
44. e Valve Replace seal Check Stator Hedo connections with new Ferrules and nuts 0060 122 21 User manual Triathlon Appendix J Logbooks The following pages contain sample pages for logbooks that can be used with the Triathlon Keep a record of settings System Menu and of programmed methods and templates Methods Menu for future reference Use copies of the provided pages 0060 123 21 85 User manual Triathlon User information Name of user Company Department Address Telephone Telefax Triathlon information Serial number Firmware version Purchase date Installed options Local dealer Service engineer Address Telephone Telefax Comments 86 0060 122 21 User manual Triathlon System Menu settings lt GENERAL gt lt USAGE gt Loop volume Protection code Timebase methods lenabled Needle tubing volume ldisabled Syringe volume Mix methods Jenabled ldisabled oyringe speed factor User program enabled Needle height disabled Skip missing vials User program protection code Air segment lyes Labeled vials enabled Headspace pressure Jyes disabled Time display HH MM SS H MM mm Templates Jenabled disabled Key click Jon loff Error beep Jon loff Calibration vials enabled Alarm buzzer Jon loff disabled lt TRAY gt lt IO gt type number Inject marker pulse length type number typ
45. e computer pin 3 RD Received data from the computer pin 5 SG Signal ground Also indicated as GND in some devices 50 0060 122 21 User manual Triathlon Appendix A Installation The Triathlon shipping container is supplied with a packing list Please check that all items mentioned in the list are included in the package before you start the installation procedure for the Triathlon Do not install the Triathlon in areas subject to shock dust or in direct sunlight Do not place it near a source of heat as this will disturb tray cooling if this option is installed The Triathlon must only be connected to power sources and apparatus with protective grounding Execute the following steps to install the Triathlon 1 Lift the Triathlon from the shipping container Make sure you keep the apparatus upright lift it by placing your hands under the Triathlon 2 Place the Triathlon 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 Install tray figure 1 1 number 6 in the Triathlon Check whether local voltage matches voltage indicated on back panel of the Triathlon Connect the power cord to the Triathlon figure 1 2 number 5 Switch the Triathlon on by using the switch at the back of the apparatus figure 1 2 number 4 The Triathlon now starts The display indicates that
46. eason it is important to make sure that the application was running without problems before and that no changes have been made in the settings System Menu 82 0060 122 21 User manual Triathlon NO Blockage Disconnect INJECTION in Flow needle from Path Valve Start Solvent Check manual flowing out of Yes as needle wash inj port i NO Disconnect Buffer tubing from valve Start Solvent Bisckeretor manual out of open Yes seal wash end NO Disconnect Buffer tubing from Syringe valve Start Solvent Check manual out of syringe Yes buffer wash valve tubing No v Leakage inside Injection Valve Check for over tightened connections in the entire Flow Path amp Check the syringe valve Disconnect the needle tubing and the buffer tubing Observe ports 3 and 4 for leakage Check rotor Yes seal Connect port 1 to an HPLC pump Block port 6 Start the pump at a low flow Recheck with manual valve 0060 123 21 83 User manual Triathlon BAD REPRODUCIBILITY Air in Flow Path v Leaking Syringe v Leaking Syringe Valve Rotor seal worn out 84 Dead volumes in tubing connections Start manual wash Replace plunger tip or Syringe Replac
47. ection method Series 1 Number 1 Number Type partial loopfill Injection method Analysis time 1 00 min Wash method Flush volume 35 ul Mix method Injections vial 3 First vial Injection volume 1 10 ul Last vial Injection volume2 10 ul First destination vial Injection volume 3 10 ul Reagent A vial Mix method 1 Step Action 1 Aspirate Sul Air 2 Aspirate 225 uL Reagent A 3 Dispense 180 uL to Destination 4 Dispense 50 uL to Waste 5 Repeat last 4 steps 1 time 6 Aspirate 5pgL Air 7 Aspirate 60 uL Sample 8 Dispense 40 uL to Destination 9 Dispense 50 uL to Waste 10 Aspirate 50 uL Air 11 Aspirate 200 uL from Destination 12 Dispense 200 uL to Destination 13 Repeat last 2 steps times 14 End of mix method Example chromatogram of the mixing and carry over test 30 0 46 Response MilliVolts 0 0 1 0 66 1 82 3 18 3 0 4 0 5 0 Time Minutes None 01 01 03 02 Speed O cO C0 Ii O1 CO T TO 6 0 Series 2 Number Injection method Wash method Mix method First vial Last vial Height 02 02 02 02 20 1 none none A 01 A 02 0060 122 21 Peak rea number 38802 38968 39088 395258 394090 394471 2 3 4 5 6 7 8 9 User manual Triathlon From the integration results the RSD for the two vials and the carry over for the Triathlon can be calculated For this example the results are RSD vial A01 sample 0 15 RSD vial A03 1 10 di
48. ed flush volume in case an injection 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 see also the full loop injections 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 and will not be injected Use of an air segment can be enabled in the System Menu General Menu Needle njection loop Buffer tubing A Sample Eluent Flush Air B Somple Eluent Flush with air segment without air segment H 3 uL pick up injections The switching sequence for a UL pick up injection is schematically shown in the following figures The initital situation the injection valve is in INJECT position The sample needle has entered the vial of transport liquid mobile phase to avoid disturbance of the co Synge am Chromatogram with an additional peak of the transport solvent after the air needle has prepunctured the septum The headspace pressure applied through the outer air needle ensures that no air or vapor bubbles are formed during wash solvent aspiration Transport liquid Sample For the first injection after wash
49. filled before starting new series Start Stop to start the Triathlon 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 Triathlon is ready for the following next run 26 0060 122 21 User manual Triathlon Example 3 1 10 dilution followed by 10 pL partial loopfill injection This example describes how to let the Triathlon transfer 360 UL from Reagent A to the destination vial add 40 uL of sample mix 3 times with 250 uL and subsequently inject 10 LL For system settings System to enter the System Menu lt USAGE gt E to enter the Usage Menu to go to the Mix field lt ENABLED gt to enable use of mix methods Escape Escape to return to the Ready Menu As soon as 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 To program the injection method Description Methods to enter the Methods Menu INJECTION 03 E to enter the Injection Menu PARTIAL E to select partial loopfill injection mode 100 E to define the analysis time 50 E to define the flush volume 3 E to define the number of injections per vial 10 E to enter the injection volume for 1st injection 10 E to enter the injection volume for 2nd injection 10 E to e
50. 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 njection loop Buffer tubing A Transport Sample Air Eluert Transport Adr B Transport Sample Eluent Transport UL pickup injections with or without air segment In case of UL pick up injections there will be no air pressure headspace pressure on the sample vial to prevent errors due to air expansion during switching from sample vial to transport vial You are advised to switch off use of an Air segment in the System Menu if the UL pick up injection method is used 80 0060 122 21 User manual Triathlon Appendix I Trouble shooting Start up problems The instrument does not work after switching on of power The instrument is working but display doesn t work No response from keyboard but display works Error code Power cord unplugged or faulty Plug in Yes replace power cord No Blown fuse Yes E Replace fuse NO Defective electronics Call your service representative Display Call your component s service failure representative Keyboard Call your component s service failure representative
51. ge actions have been counted Syringe valve will have to be replaced and the counter for valve actions will have to be reset by the maintenance engineer DEFAULT ALL to change all software settings to default All series methods templates and the user program unless protected by protection code will be erased If DEFAULT ALL is selected check whether hardware configuration still is compatible with settings entered in the System Menu lt SSV gt option Use this key to start a procedure in which all lines of the solvent selection valve can be primed The menu offers the following possibilities lt SSV1 gt to to open the corresponding port of the solvent selection valve lt SSV6 gt lt PRIME gt to prime the selected solvent line with one syringe volume The last selected port will remain active after leaving the SSV mode lt COOL gt option Use this key to enter the programming mode for Peltier tray cooling The programmable temperature range is 4 C to 40 C The maximum cooling capacity is approximately 20 C below ambient refer to Appendix B for specifications Connect the condensed water and leakage connector to a waste container on the floor to drain condensed water If the cool option is switched lt ON gt the following soft function keys can be selected lt MANUAL gt temperature control will remain OFF until it is switched on again by the user in this menu lt AUTOMATIC gt temperature control wi
52. ge next vial Freeze input active Reset outputs after series lt TIMEBASE gt MIX Mix method e ASPIRATE sample destination air reagent DISPENSE Timebase program 4x AUX 1 4x AUX 2 4x AUX 3 4x AUX 4 4 x ISS B 8x SSV 8x CODE OUT END TIME sample destination waste reagent REPEAT WAIT WASH LJ LJ e ISS A LJ LJ LJ LJ Note These parameters depend on the contents of the methods used and or the System Settings of the Triathlon Note Optional Soft function keys PRIORITY Only available when user program and mix method have been disabled in the System Settings 73 User manual Triathlon Ready Menu lt Menu gt lt ADVANCE gt lt WASH gt lt SYR END gt lt UTILS gt lt SERIAL gt lt SSV gt COOL SERVICE e Cool ON OFF lt COPY lt ERASE gt LOG lt DEFAULT ALL e Setpoint Switch ON OFF Advance tray Perform Move syringe method Erase methods Logbook All methods Communication Prime solvent Service mode standard wash amp Userprog erased all settings protocol valve to default System Settings System Menu gt GENERAL USAGE lt CLOCK gt lt COMM gt Loopvolume Method protection code 1 type number Inj Marker pulse e Device identifier Needle tubing Use time base method 2 type number Vial marker pulse Syringe volume Use mix method 3 type number Lab Vial marker pulse S
53. gt 500 uL WASH Use this key to program wash methods It is possible to program a wash between injections samples or series For each wash method the volume of wash solvent can be defined The minimum programmable volume is 300 LL lt TIMEBASE gt if enabled in System Menu Usage Menu Press this key to enable control of the optional ISS valve and other devices via auxiliary or binary outputs A maximum of 5 timebase methods can be programmed The menu offers the following soft function keys lt AUX gt scroll through all program lines by pressing E or select AUX to move to the next auxiliary lt VALVES gt controls the ISS valve and the solvent selection valve The ISS valve can only be programmed if the optional ISS valve is installed 6 1 and 2 1 refer to the interconnected ports of the valves Press E to scroll through programming lines Only if SSV option is installed Enter the time and the SSV port number value between 1 and 6 0060 123 21 39 User manual Triathlon Figure 5 2 ISS valve interconnected ports lt CODE gt outputs enter a time and a value between1 and 15 hexadecimal output Press E to scroll through the programming lines lt END gt 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 pressed the Triathlon will automatically generate an end time The end time is equal to the analysis time programmed in the in
54. he 250 mL wash solvent bottle 1 wash solvent Use of methanol organic solvent solvent degas it with Helium or an ultrasonic figure 2 1 Fill the wash solvent bottle with the appropriate Injection valve no buffers or a mixture of water and isopropanol 80 20 is recommended Before using the wash bath V ocrew the bottle to the cap in the holder Place the holder in the Triathlon as indicated in Put the wash solvent tube in the wash solvent Lower the cover of the Triathlon Select soft function key SYR END in the Ready Menu to fill the syringe Place the syringe back in home position by selecting soft function key lt SYR HOME gt Fiaure 2 1 Wash solvent bottle Repeat steps 6 and 7 until the wash solvent tube and the syringe are completely filled Select soft function key WASH to perform a standard wash routine Waste I any air remains in the syringe select lt SYR END again to fill the syringe with wash solvent press lt SYR again to move contents to waste Repeat if there is still air in the syringe and gently tap the syringe as wash solvent is dispensed to waste 0060 122 21 User manual Triathlon If you use an application that requires more than 250 mL of wash solvent for a complete run install a longer tube with flanged end for valve fitting and place a larger bottle next to the Triathlon To fill the wash solvent tube you may have to repeat the ab
55. injections from the same vial If a wash between injections has been programmed sample loss is 30 uL for every injection For zero sample loss injections use the uL pick injection mode e Pick up use a 100 uL syringe for optimum accuracy and reproducibility Do not use a smaller sample loop The sample plug is transported into the loop preceded by a programmable air segment of 5 UL if selected in System Menu you are advised to switch off the air segment with uL pick up with a plug of transport liquid which equals 2 5 times the programmed needle tubing volume 12 0060 122 21 User manual Triathlon Injection volumes up to twice the standard With the standard 250 uL syringe standard needle with tubing 15 ul and standard 500 uL buffer but with a 200 uL sample loop the maximum injection volumes are Flushed loop 200 uL sample loss remains 230 uL since loops gt 100 uL need only one loop volume overfill 30 UL pre flush Partial loopfill 100 ul uL Pick up 77 uL Volumes smaller than 5 uL may be injected but reproducibility and accuracy may not be lt 0 5 for partial loopfill or lt 1 for UL pick up In short loop lt 100 uL loop has to be filled three times loop gt 100 UL 499 uL loop has to be filled twice loop gt 500 uL loop has to be filled 1 5 times For volumes larger than 200 pL Use the 2000 uL buffer use the appropriate sample loop size and the appropriate syringe Syringe volume gt 2 x injection v
56. ition range 1 to 6 Table 5 3 Syringe speed SYRINGE 100 uL 250 uL 500 uL 1000 uL mE 50 uL min 125 uL min 250 uL min 500 uL min 0 5 mL min 2 low 125 uL min 315 630 uL min 1255 uL min 1 3 mL min 3 normal 250 uL min 625 uL min 1250 uL min 2495 uL min 2 5 mL min 4 high 375 uL min 940 uL min 1880 uL min 3765 uL min 3 8 mL min 770 uL min 1920 uL min 3840 uL min 7680 uL min 7 7 mL min 1070 uL min 2675 5335 10665 uL min 10 7 mL min 7 1370 uL min 3430 uL min 6855 uL min 13710 uL min 13 7 mL min 1745 uL min 4365 uL min 8725 uL min 17450 uL min 17 5 mL min 2135 uL min 5335 uL min 10670 uL min 21335 uL min 21 3 mL min During the dispense action the pressure in the buffer tubing will increase To prevent damage of the buffer tubing the flow should not exceed the value of CAUTION 6 mL min for water Maximum speed 9 for 100 uL and 250 uL syringes speed 6 for 500 uL syringe and speed 4 for 1000 uL and 10 mL syringe If more viscous liquids are used the speeds should be reduced 42 0060 122 21 User manual Triathlon 5 4 Series Menu This menu allows you to define the run sequence in series maximum of 24 series can be programmed series contains information about the methods to be used for a range of vials This can be a template a separate method mix injection wash timebase or the user program Information on location of vials labeled vials or calibration vials is als
57. jection method used in the same series If end time exceeds the programmed analysis time this end time overrules the analysis time It is possible to program events after the end time but these events are not carried out during a run lt USER PROGRAM gt The user program offers the possibility to program all possible actions required for a sample handling sequence in separate steps Note that the total number of 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 soft function keys appear e EDIT an existing step or program a new step for the user program e lt INSERT gt a new step before the displayed step e DELETE delete the displayed step The edit and insert menus offer the following soft function keys lt ASPIRATE a programmed volume from sample vial ambient air destination vial wash or one of the reagent vials into the buffer tubing Speed and height of syringe can be entered refer to table 5 1 page 34 The maximum volume that can be aspirated is the total volume of the syringe lt DISPENSE gt a programmed volume from the buffer tubing into the sample vial waste destination vial wash or one of the reagent vials Speed and height of sy
58. k the numbers of methods to the template The following items can be entered to fill a template e user program instead of methods if soft function key lt YES gt is selected the complete template is filled with the user program no other methods can be added If soft function key lt NO gt is selected the template can be filled with the following e mix method number injection method number wash method number timebase method number A maximum of 24 templates can be programmed 0060 123 21 37 User manual Triathlon lt METHODS gt Use this key to enter a menu in which methods can be defined lt MIX gt if enabled in System Menu Usage Menu Use this key to program a method that allows you to perform pre injection sample handling e g pre column derivatization dilution or adding of internal standard Nine mix methods can be programmed the maximum number of steps that can be programmed for the total of 9 mix methods and the user program is 240 Assign a number to the mix method The Mix Menu appears e EDIT an existing step or a new step for a new mix method e INSEHT a new step in an existing method before the displayed step e DELETE the displayed step 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 e
59. layed System to enter the System Menu in which system settings can be entered e General keys Escape allows the user to leave the progamming mode or go to a previous level in the menu Entered values are checked for validity and then saved Help to display help information available only for a limited number of functions 0060 123 21 5 User manual Triathlon 6 0060 122 21 User manual Triathlon Chapter 2 Preparations for use This chapter describes procedures for replacement or installation of parts either for maintenance or for preparation of the Triathlon for an analytical run Please note that the power should not be switched off for any of the procedures described below as soft function keys in the Ready Menu must be used number of items required for use of the Triathlon are factory installed see Appendix However after the installation procedure described in Appendix has been executed check that the following have been correctly installed before you start to use the Triathlon e HPLC pump amp column connections e waste tubing e wash solvent bottle e syringe e needle assembly e syringe sample loop and buffer tubing e tray segments and tray types e reagent vials transport vials Loosen the screw at the right hand side of the cover and lift the cover to be able to execute the procedures described in this chapter 2 1 HPLC connections Make the following connections e HPLC pump to port
60. le 4 5 lt PROG OUT gt 41 program method 24 25 27 28 programming keys 5 0060 122 21 programming options 73 74 protection code 29 30 35 R 21 Ready Menu 19 31 33 51 reagent vials 7 reference 31 44 lt REMOTE gt 21 remote control 21 49 lt REPEAT gt 38 replacement of parts 7 18 reproducibility 1 12 13 63 64 reset outputs after last series 37 Rheodyne valve 2 69 71 RS232 3 33 RS422 485 3 run control keys 4 run series 21 24 26 29 30 safety practices iii sample handling 20 sample loop 11 13 sample loss 1 12 13 sampling needle 10 11 scale factor 33 sensor 10 lt SERIAL gt 33 Series 5 19 20 executing 21 in remote control 21 Series Menu 19 43 44 parameters 43 without templates 43 44 with templates 43 44 lt SERVICE gt 33 settings see System Menu silica coated needle 11 skip missing vials 34 soft function keys 4 specifications 53 55 lt SSV gt 32 42 lt START gt 21 24 Start Stop 4 stop I O 49 symbols i ii lt SYR gt 41 lt SYR END gt 8 9 31 lt SYR HOME gt 8 9 29 0060 123 21 User manual Triathlon syringe 2 9 11 13 calibration 67 replacing 9 types of 11 waste 8 syringe speed 33 35 42 SYH VALVE 41 syringe volume 33 syringe waste tubing 2 System 5 19 oystem Menu 19 23 25 27 29 29 33 37 T Tefzel tubing 53 temperature control 32 TEMPLATE 30 37 template 20 create 29 example 29 30 enable disable 35 lt TIMEBAS
61. ll be switched OFF after all programmed series have been executed lt DATE TIME gt temperature control will be switched OFF at a date and time that can be programmed 32 0060 122 21 User manual Triathlon lt SERIAL gt Use this key to put the Triathlon in serial mode to allow for control of the autosampler by way of PC RS232 interface Select a device identifier in the System Menu refer to section 5 2 method protection code was defined in the system settings this code must be entered to get access to serial mode The following soft function keys appear lt PANIC gt press this key to begin a stop sequence in which all tubing is rinsed and the valve and 1 0 ports are reset At the end of the sequence serial mode is resumed lt EXIT gt press this key to end serial mode and return to the Ready Menu lt SERVICE gt For service to the apparatus To be used by authorized personnel only The Service Menu is protected by a service code 5 2 System Menu The System Menu contains the following soft function keys lt GENERAL gt Press this key to enter values for e loop volume needle tubing syringe volume these values have to be entered because the Triathlon can be fitted with various types of syringes Every needle and needle tubing volume requires a different minimum flush volume The default flush volume equals two times the volume of needle and tubing is enabled in the System Menu Usage Menu When the 10 mL sy
62. loop 20 UL Rheodyne loop 50 uL 0060 122 21 number 3 96 012 3796 013 3796 014 3796 015 3796 016 3796 076 3796 032 3796 033 3796 040 3796 041 3796 042 3796 050 3796 062 3796 065 4340 025 4400 000 4400 030 4400 250 4400 500 4400 251 4400 001 4400 601 4400 602 4400 603 4400 604 4400 607 4400 605 4400 606 0060 123 21 User manual Triathlon description Rheodyne loop 100 uL Rheodyne loop 200 uL Rheodyne loop 500 uL Rheodyne loop 1000 UL Rheodyne loop 100 uL Peek Rheodyne loop 20 ul Peek Rheodyne Rheflex ferrule Rheodyne Rheflex nut Rheodyne 9010 Rotor seal Rheodyne 9010 Stator Rheodyne Stator face assembly Rheodyne 7010 Tefzel rotor seal Flanged tubing fitting 1 8 Flanged tubing fitting 1 16 Wash solvent bottle 250 mL Syringe 1000 uL Syringe100 uL syringe 250 uL syringe 500 uL Syringe 25 uL for micro option Triathlon oyringe 10 mL Plunger replacement tip 100 uL pck 10 Plunger replacement tip 250 uL pck 10 Plunger replacement tip 500 uL pck 10 Plunger replacement tip 1000 uL pck 10 Plunger replacement tip 25 uL pck 10 micro option Plunger replacement o ring 500 uL pck 10 Plunger replacemnt o ring 1000 uL pck 10 gt 61 User manual Triathlon 62 0060 122 21 User manual Triathlon Appendix E Calibration and performance The Triathlon is factory tested for reproducibility carry over and mixing according to the following test procedure 1 An
63. lution 0 37 96 1 10 dilution 1 10 13 variation 1 29 The specified variation is 5 Carry over vial A02 0 296 The real value cannot been calculated the specified value is a Carry over 0 296 E 2 Syringe calibration To calibrate the syringe dispense a volume of 200 uL water from a sample vial to an empty destination vial with the following mix method Mix method 1 Step Action Speed Height 1 Aspirate 250 uL Sample 2 02 2 Dispense 200 uL to Destination 3 02 3 End of mix method Series 1 Injection method None Wash method None Mix method 1 First sample vial A 01 Last sample vial A 01 First destination vial A 02 0060 123 21 Weigh the destination vial before and after the Run The difference is the dispensed volume The specified variation is 296 67 User manual Triathlon E 3 Tray cooling calibration p Place a thermocouple on the bottom of the tray make sure the contact is good Switch on the tray cooling and program setpoint of 10 Wait minimal 15 minutes for equilibration of the Triathlon Read out the temperature of the thermocouple The value must be in a range of 2 C of the programmed setpoint E 4 Loop calibration QUE eee We Disconnect the loop from the injection valve Remove all liquids from the loop with air Weigh the empty loop on an analytical balance Fill the loop with minimal 2 times its volume of water Weight the filled loop again The differe
64. ly open Vial marker Normally open STOP Common Vial marker Common STOP I O Normally closed Vial marker Normally closed 24 V DC Labeled vial marker Normally open Power ground Power ground Vmax 28 Voc Vac 0 25 0060 123 21 45 User manual Triathlon Table 6 3 Connector P5 AUXILIARIES AUX 1 Normally open AUX 3 Common AUX 1 Common AUX 3 Normally closed AUX 1 Normally closed AUX 4 Normally open AUX 2 Normally open AUX 4 Common AUX 2 Common AUX 4 Normally closed AUX 2 Normally closed 24V DC AUX 3 Normally open Power ground Power ground Vmax 28 Voc Vac 0 25 Maximum current for 24 Voc supply is 0 5 A total Normally closed NC Common COM Normally open NO Figure 6 1 Contact closures outputs 6 2 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 a connection diagram is shown in figure 6 2 page 45 Table 6 4 Connector 2 TTL OUTPUTS INJECT MARKER D 4 BCD or HEX 10 or 16 VIAL VIAL MARKER D 5 BCD or HEX 20 or 32 LABELED VIAL MARKER D 6 BCD or HEX 40 or 64 STOP D 7 BCD or HEX 80 or 128 D O BCD or HEX 1 Signal ground D 1 BCD or HEX 2 Signal ground D 2 BCD or HEX 3 Signal ground D 3 BCD or HEX 4 All markers are active low logical 0 Vmax 5 5 V l
65. miliar with standard laboratory and HPLC terminology Chapters 1 and 2 of this manual contain basic information that should be read by every user of the Triathlon e Chapter 1 identifies main parts of the Triathlon e Chapter 2 explains how to install parts either for maintenance purposes or for preparing the Triathlon for an analytical run For first time users e Chapter 3 explains the main menus of the Triathlon and offers a basic explanation of how to operate the Triathlon e Chapter 4 offers a number of examples that teach the user how to work with the Triathlon For experienced users e Users who understand how the Triathlon works will probably only use the reference part of this manual Chapter 5 to look up the purpose of a particular function e Experienced users may find the overview of programming options Appendix useful for quick reference of possibilities of the Triathlon The appendices in this manual offer specialist information An index has been provided to allow the user to find required information quickly Typographical conventions The following typographical conventions are used in this manual Initial Capitals are used for names of the menus Ready Menu Series Menu etc lt CAPITALS gt are used to refer to soft function keys bold type is used for names of keys in the keypad 0100 is used to indicate values that have to be entered with the numerical keypad 0060 123 21 User manual Triathlo
66. mpatible with other laboratory equipment e apparatus designed for routine analysis and method development In addition options like tray cooling and switching valves are available for the Triathlon These options enhance the uses for the Triathlon considerably refer to Appendixes B and D for more information on options 1 1 Injection principle The Triathlon offers three different methods of injection for an analytical run Flushedloop The sample loop is completely quantitatively filled with sample resulting in extremely good reproducibility Partial loopfill The sample loop is partially filled with sample this means low sample loss and programmable injection volumes UL pick up After aspiration of sample the sample is transported into the loop with transport liquid mobile phase this means no sample loss The Triathlon uses a syringe to aspirate the sample from a vial into the sample loop To prevent contamination of the syringe the Triathlon is equipped with a buffer tubing between the syringe and the injection valve Wash solvent is used to remove the sample from the buffer tubing and sample needle and to rinse the buffer tubing and sample needle For more technical information on the injection principle used by the Triathlon refer to Appendix H For an overview of fluid connections of the Triathlon refer to the illustration inside the cover of the Triathlon 0060 123 21 1 User manual Triathlon 1 2 Front view of the Triathlon
67. mple will be added and after mixing 3 times a 10 uL injection will be performed Example 4 Defining a template and adding a protection code This examples describes how to incorporate the injection method 02 and wash method 01 defined in example 2 in a template A protection code will be added For system settings System To enter the System Menu lt USAGE gt E To enter the Usage Menu 123456 E To enter a 6 digit code memorize this code 0060 123 21 29 User manual Triathlon E to go to the mix methods field lt DISABLED gt to disable use of mix methods to go to the template field lt ENABLED gt to enable templates Escape Escape to return to the Ready Menu After use of templates has been enabled the message ALL SERIES DEFAULT appears The user will have to redefine series because the settings have been changed To select the methods to be incorporated in the template 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 template 01 to define the wash method for this template Escape Escape to return to the Ready Menu To define the series Series to enter the Series Menu 01 E to define the Series number 01 E to define the Template method number 01 E to define the first sample vial 02 E to define the last sample vial
68. n Symbols The following symbols are used on the Triathlon WARNING This sticker indicates that care should be taken to prevent N personal injury or damage to parts of the Triathlon SHARP NEEDLE AND MOVING PARTS KEEP HANDS CLEAR AN The following pictograms are used in this manual A AN This sticker with yellow background color at the back of the Triathlon calls attention to the fact that you are expected to consult this manual for instructions on how to operate the Triathlon calls attention to a procedure which if not correctly executed could result in injury or loss of life Do not proceed beyond a DANGER sign until the indicated conditions are fully understood and met calls attention to a procedure which if not correctly executed could result in personal injury Do not proceed beyond a WARNING sign WARNING until the indicated conditions are fully understood and met calls attention to a procedure which if not correctly executed could result in damage to the equipment Do not proceed beyond a CAUTION CAUTION sign until the indicated conditions are fully understood and met gt calls attention to important information Read this information before continuing i 0060 123 21 User manual Triathlon Safety practices The following safety practices are intended to ensure the safe operation of the equipment A Removal of panels may expose users to dangerous voltages Disconnect the
69. nce in weight is the capacity of the loop The weight divided by the specific weight of water 1 g mL gives the calibrated volume of the loop Allowed variation 10 According to Rheodyne 68 0060 122 21 User manual Triathlon Appendix F Rheodyne injection valve The Triathlon is standard equipped with a Rheodyne 7739 valve the biocompatible Triathlon is equipped with a Rheodyne 9740 valve The text below is a copy of the Rheodyne Operating instructions F 1 Operating Instructions for Rheodyne Model 7739 Valve Shaft Assembly Shaft Assembly Rotated Rotated Clockwise Counterclockwise Figure F 1 Flow diagram of Model 7739 as viewed from stator Model 7739 is a two position six port stainless steel valve Figure F 1 shows the flow diagram of the valve The six small circles represent the ports in the valve stator The slots are the connecting passages in the rotor seal Rotation of the valve 60 switches the valve from one position to another Shaft Assembly Body Specifications Maximum temperature is 100 C The valve is set to hold 345 bar 5000 psi The wetted surfaces are stainless steel and an inert polymer Rotor Pins Isolation Seal e ea Rotor Seal Stator Ring Maintenance FR sate With normal use the valve will give many tens of Location Pin thousands of cycles without trouble The main cause of early failure which is seen as a leak in the valve is abrasive par
70. nt of flush volume This air segment is at the front of the flush volume and will not be injected and therefore will not influence the injection Use of an air segment can be enabled in the System Menu General Menu 76 0060 122 21 User manual Triathlon With a standard needle the flush volumes must be minimal 30 uL for injections with air segment and 35 uL for injections without air segment If the samples are highly viscous it may be necessary to program larger flush volumes and reduce the syringe speed for better performance Needle njection loop Buffer tubing A Sample Eluent Flush Air 2022222222222 7222 gt B Sample Eluent Flush 22 Z 7222 with air segment without air segment H 2 Partial loopfill injections The switching sequence for a partial loopfill injection is schematically shown in the following figures The first three steps are identical to those for Full loop injections see H 1 For partial loopfill injections the sample loop is filled by transporting the programmed injection volume into the sample loop The injection valve switches into the INJECT position The sample loop is now part of the HPLC mobile phase flow path the sample is transported to the column The gt analysis time starts Sample vvs H 0060 123 21 77 User manual Triathlon The next injection sequence will start with a flush of 50 of the programm
71. nter the injection volume for 3rd injection Escape to return to the Methods Menu 0060 123 21 27 User manual Triathlon To program the mix method lt MIX gt to enter the Mix Menu 1 E to define Mix method number 1 lt INSERT gt to define mix method step number 1 lt ASPIRATE gt 5 AIR E to aspirate an air segment of 5 uL INSERT to define mix method step number 2 ASPIRATE 225 to aspirate 225 UL Menu lt REAG A gt E to perform defined asp From reagent vial A lt INSERT gt to define mix method step number 3 lt DISPENSE gt 180 to dispense 180 uL to gt gt 02 destination vial lt INSERT gt to define mix method step number 4 lt DISPENSE gt 50 to dispense 50 uL WASTE 5 E to dispense defined volume to waste INSERT to define mix method step number 5 REPEAT 1 4 E to repeat last four steps once lt INSERT gt to define mix method step number 6 lt ASPIRATE gt 5 AIR to aspirate an air segment of 5 uL lt INSERT gt to define mix method step number 7 lt ASPIRATE gt 60 lt SAMPLE gt to aspirate 60 uL of lt INSERT gt sample lt DISPENSE gt 40 to define mix method step number 8 lt DESTINATION gt gt gt 2 E to dispense 40 uL lt INSERT gt to destination vial DISPENSE 50 lt WASTE gt to define mix method step number 9 gt 5 E to dispense 50 uL lt INSERT gt to waste lt ASPIRATE gt 50 lt AIR gt to define mix method step number 10 lt IN
72. nu appears after the Methods key in the keypad has been selected The menu allows you to program a method to be used in an analytical run and to assign a number to it e Series Menu this menu appears after the Series key in the keypad has been selected The menu allows you to define a series and to assign a number and a method to it for the analytical run Use the following keys to go through the menus E ENTER to confirm a choice or to select a choice made in a screen or to step through menu lines cursor keys to change values in a field or to move to a different field in a screen Escape to return to a previous menu CL to remove a value from a field and enter NONE or DEFAULT soft function keys labels displayed in bottom line of the screen to go to submenus If MENU or MN is displayed in the top right hand corner of the screen you can press the Menu key in the keypad to display more possibilities offered by the menu Refer to section 1 4 for an explanation of all keys in the keypad 0060 123 21 19 User manual Triathlon 3 2 Convenient working order After you have determined what type of analytical run you want to perform the most convenient working order for the Triathlon is 1 Enter settings in the System Menu Note that the settings for the Triathlon probably already have been correctly entered factory installed 2 Program a method for the analyses you wish to perform in the Methods Menu 3 Define a series and link
73. o programmed in a series The settings entered in the System Menu and the methods defined in the Methods Menu determine which possibilities appear in the Series Menu Table 5 4 Series parameters Without templates With templates Use user programYes No Template number Injection method number e 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 Use calibration vials Yes No O First calibration vial O Last calibration vial O No of samples between calibration Calibration vials are only available if enabled in the System Menu not available if Mix Method has been programmed First sample vial e Last sample vial Only if a mix method has been programmed or if used in user program O First destination vial O Vial Reagent A O Vial Reagent B O Vial Reagent C O Vial Reagent D Only if the use of labeled vials has been enabled in the System Menu O Labeled vial no 1 O Labeled vial no 2 O Labeled vial no 3 O Labeled vial no 4 e marked questions are always asked in series O marked questions depend on the used methods and the settings entered in the System Menu 0060 123 21 43 User manual Triathlon Explanation 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 5 4 gives an
74. ogical 1 gt 3 5 V logical 0 lt 1 0 V DC output source sink current 20 mA switches from LOAD to INJECT However in a User Program markers have to be programmed by the user marker output pulse will be generated when the injection valve 46 0060 122 21 User manual Triathlon BCD Binary Coded Decimal The BCD output consists of a 2 digit vial number 03 00 represent the single digits 0 9 and D7 D4 represent the tens Table 6 5 BCD output conversion Table 6 6 Examples of BCD vial number output Sngedgi Vial number Output 96 9 x 10 107 6x1 Dee Output 59 1 50 9 Output 0 0 10 1 10 10 10 0 0060 123 21 47 User manual Triathlon Hex Hexadecimal Code The hexadecimal output represents the vial number as 8 bits by assigning values of 1 2 4 8 16 32 64 and 128 to every single bit DO D7 Table 6 7 Examples of hexadecimal output D7 D6 D5 D4 gt a Vial 128 64 32 16 number Output 128 16 4 2 Output 32 16 8 2 1 Table 6 8 Connector P3 TIMED OUTPUTS 4 bit time base code output 1 1 6 Signal ground 2 TB1 HEX 2 7 Signal ground 3 TB2 HEX 4 8 Signal ground 8 9 Signal ground 4 HEX 5 not used Vmax 5 5 V logical 1 gt 3 5 V logical 0 lt 1 0 V DC output source sink current 20 mA Autosampler Ext device Output 7 Input TTL NES GN GND
75. olume Injection volumes larger than 500 uL are possible but the sample may contaminate the syringe Program sufficient wash after use Overview of appropriate buffer tubing for each type of syringe Syringe Buffer tubing 500 uL 500 uL 2000 uL 2000 uL 15 mL The 10 mL syringe and the 15 mL buffer tubing can only be used in combination with the User Program and not with the standard injection methods 0060 123 21 13 User manual Triathlon 2 8 Tray segments and Vial types A wide range of vials and septa can be used for the Triathlon from micro vials 0 5 mL to super LSV vials 10 mL The volumes of standard vials can be reduced by using inserts The Triathlon can be equipped with four types of tray segments to accomodate use of the various types of vials see figure 2 4 Type A Standard tray segment Type B LSV tray segment Type C Super LSV tray segment Type D Micro vial tray segment Figure 2 4 Types of tray segments The Triathlon allows you to use more than one type of segment in a tray Enter the configuration of the tray in the System Menu Tray Menu The following vial types can be used Chromacol vials for standard tray outer vial diameter 12mm 1 8 ml 1 8 ml 1 8 ml 2 5 ml 2 5 ml C ee Dc D EN 20006 29 Figure 2 5 Chromacol vials 14 0060 122 21 0 6 ml User manual Triathlon
76. ons The Triathlon has six standard I O connectors on the rear side five OUTPUT connectors and one INPUT connector Refer to figure 1 2 for location of connectors The communication connector is a standard RS232 or an RS422 485 communication interface connector The configuration of the I O connector is described in the tables below The manufacturer will not accept any liability for damages directly or indirectly caused by connecting the Triathlon to instruments which do CAUTION not meet relevant safety standards 6 1 Contact closures outputs The following three tables with programmable outputs P1 marker outputs P4 and auxiliary outputs P5 are contact closures outputs floating NO NC contact see figure 6 1 page 44 Table 6 1 Connector P1 OUTPUTS 2 programmable outputs and alarm output OUT 1 Normally open Spare OUT 1 Common Spare OUT 1 Normally closed Alarm output Normally open OUT 2 Normally open Alarm output Common OUT 2 Common Alarm output Normally closed OUT 2 Normally closed 24 V DC Spare Power ground Power ground Vmax 28 Voc Vac 0 25 The Alarm output will be activated whenever error occurs refer to Appendix C for a description of the error codes of the Triathlon Table 6 2 Connector PA MARKERS Inject marker Normally open Labeled vial marker Common Inject marker Common Labeled vial marker Normally closed Inject marker Normally closed STOP 1 0 Normal
77. ove mentioned filling procedure steps 6 and 7 a few times 2 4 Syringe The Triathlon is supplied with a 250 uL syringe It is also possible to use the Triathlon with a 100uL 500 uL 1000 uL or 10 mL syringe Execute the following steps to install a syringe 1 Select soft function key lt SYR END in the Ready Menu to move the syringe to end position 2 Lift the cover ed Unscrew the top of the syringe turn clockwise 4 Pull the bottom of the syringe towards you you can now remove the syringe refer to figure 2 2 5 Fill the new syringe with wash solvent and make sure that all i i Q IO air bubbles are removed from the syringe M 6 Connect the bottom of the filled syringe to the Triathlon t 7 Screw the top of the filled syringe to the Triathlon counter clockwise AT 8 Lower the cover 9 Select soft function key lt SYR HOME gt to remove air from the syringe The syringe moves to home position and its A contents is dispensed to waste E 10 If any air remains in the syringe select lt SYR again to fill the syringe with wash solvent press lt SYR HOME gt again to move contents to waste Repeat if there is still air in the syringe and gently tap the syringe as wash solvent is dispensed to waste 11 Select soft function key WASH in the Ready Menu to execute a standard wash routine All tubing connected to the syringe valve is filled and
78. overview of the items you have 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 vial and the last sample vial 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 vial and last sample vial If you have enabled use of calibration vials in the System Menu Usage Menu you will have to define whether you will use calibration vials and indicate the location of the first and last calibration vial and indicate the number of vials between calibration vials refer to figure 5 3 However if you have for example enabled use of a Mix Method in the System Menu Usage Menu you will also have to define the location of the First destination vial and Reagent vials 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 Q Series are stored in the Triathlon memory for as long as the power is on As Calibration Sample Q9 Q0 Q2 9 Q0 09 Q9 0 Q9 9 Q9 Figure 5 3 Injection sequence with 3 calibration vials between every 5 vials 44 0060 122 21 User manual Triathlon Chapter 6 I O Connecti
79. puts should be reset to default after the last series Refer to Chapter 6 for more specific information on I O connections 36 0060 122 21 User manual Triathlon Table 5 2 Overview I O menu Possibility default inject marker pulse length 1 0s 0 1 2 0 vial marker pulse length 1 0s 0 1 2 0 labeled vial marker pulse length 1 0s 0 1 2 0 input edge next injection falling falling or rising input edge next vial falling falling or rising freeze input active low low or high reset outputs after last series no yes or no lt CLOCK gt Use this key to switch the system clock on or off Select ON to enter the clock menu in which you can set date yy mm dd and time hh mm This date and time will be displayed in the Ready Menu lt COMM gt Use this key to define a device identifier for communication with other equipment e g a PC An identifier between 20 and 29 can be selected for the Triathlon 5 3 Methods Menu This menu allows the user to program various types of methods it is possible to define 24 separate injection methods 5 wash methods 5 timebase methods 9 mix methods and one user program lt is also possible to program a combination of methods and save them in a template The settings entered in the System Menu determine the possibilities offered by the Methods Menu 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 lin
80. r numerals in the various programming fields e CL to clear a value in a field or replace it by NONE or AUTO e stands for Enter to go through menu lines or to confirm a choice made in a menu or a value entered The entered value is checked for validity and then saved Function keys e control keys Start Stop to start or stop automatic processing or to reset the system after an error has occurred Hold Cont to hold or continue the analysis time The analysis time is extended by the period that Hold is active Interrupt not used Priority to stop run to process priority sample 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 priority sample is series of one vial with an injection 4 0060 122 21 User manual Triathlon method a wash method and a time base method defined in a template only possible if the correct settings are entered in the System Menu e Programming keys see also section 3 1 and Chapter 5 Series to enter the Series Menu in which series can be defined for an analytical run Methods to enter the Methods Menu in which methods can be programmed for use in an analytical run Menu this key can only be used if MENU or MN is shown in the top right hand corner of the display If pressed more possibilities of the menu are disp
81. ries Menu Default disabled You are advised to 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 lt TRAY gt Use this key to define the type of tray that will be used Four types of tray segments can be selected Type A default type type and type D numbered 1 8 see fig 2 4 After all tray types have been has been selected enter e of tranport vials A D or press CL e first transport vial enter a number e last transport vial enter a number Vials can be placed in any of the vial positions Transport vials must be placed in a continuous row IO Use this key to enter the I O configuration mode and define the following e vial output BCD or HEX to define the vial output e inject marker pulse length to define the length of the inject marker pulse e vial marker pulse length to define the length of the vial marker pulse e labeled vial marker pulse length to define the length of the vial marker pulse of the labeled vial e input edge next injection to define the edge sensitive inputs for the next injection e input edge next vial to define the edge sensitive inputs for the next vial e freeze input active to define whether the freeze input is active when high or freeze input is active when low e reset outputs after last series to determine whether the out
82. ringe is selected only the User Program can be used Use of other types of methods will be disabled The 10 mL syringe can only be selected if the use of the User Program e syringe speed and scale factor The aspirating speed of the syringe used in injection methods can be adapted depending on viscosity of samples Alternatively syringe speeds can be reduced by entering a scale factor The syringe speed will be the 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 Refer to table 5 3 page 42 0060 123 21 33 User manual Triathlon e needle height distance between the needle point and 3 reference point a few millimetres above the bottom of the sample tray Not the bottom of the vial see figure 5 1 can be programmed The value in the system settings is only used in injection methods for mix methods this value is programmable in the method itself ZIRAY BOTTOM A height of 0 is not the bottom of the tray but a reference value related to a point CAUTION a 2 5 mm above the bottom of the tray Check sample needle height with an empty vial e skip missing vials YES means that empty spaces are skipped during the run NO means that the Triathlon will stop if an empty space is encountered during the run an error code will be generated e air segment decide whether an air segmen
83. ringe can be entered refer to table 5 1 page 34 It is not possible to dispense a larger volume than the total volume aspirated in previous actions 40 0060 122 21 lt SYR VALVE gt lt SYR gt lt WASH gt lt VALVES gt lt WAIT gt lt COMPRES gt lt AUX gt lt WAIT IN gt lt PROG OUT gt 0060 123 21 User manual Triathlon to control the connections of the syringe to one of its three tubes NEEDLE connection to sample needle WASH connection to wash solvent bottle WASTE connection to syringe waste tubing to control the movements of the syringe LOAD the syringe with the programmed volume lt UNLOADs 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 to execute a needle wash the content of the buffer tubing is not rinsed to waste before the start of the wash The programmed volume of wash solvent is used to wash the needle at the wash position The wash position may be contaminated with the contents of the buffer tubing which may generate cross contamination To prevent contamination of the wash position program a dispense to waste action before programming a wash action to program positions of high pressure valves ISS injector valve SSV The injector valve has two positions lt INJECT gt and LOAD The 155 optional valve
84. ssembly fit the mating holes in the stator only one way Mount the stator and stator face assembly on the User manual Triathlon Body Shaft Assenbly i Isolation Seal Rotor Seal Body Location Pin Stator Ring Stator Location Pin Stator Face B Assenbly Stator Stator Support Ring AAN Stator Screws 3 Figure F 4 Exploded view of Model 9740 valve so the stator locating pin in the stator ring enters the mating hole in the stator Replace the three stator screws Tighten each an equal amount until the screws are tight 0060 123 21 71 User manual Triathlon 72 0060 122 21 User manual Triathlon Appendix G Programming options Ready Menu lt ADVANCE gt lt WASH gt lt SYR END gt lt UTILS gt lt COPY Advance tray Perform Move syringe Copy method Erase methods standard wash e System Settings System lt GENERAL gt lt USAGE gt Loopvolume Method protection code e Needle tubing Use time base method e Syringe volume Use mix method Syringe speed Use user program e Needle height User program protection Skip missing vials code Air segment Use labeled vials Headspace pressure Use templ
85. t vial A 06 Injections vial 3 Injection method 1 Injection volume 1 5 ul Wash method 1 Injection volume2 5 Injection volume 3 Sul Example chromatogram of the reproducibility test 2 42 5 28 8 15 11 03 13 91 16 79 19 69 22 60 25 50 28 36 31 22 34 08 36 96 39 84 42 72 45 63 48 55 51 46 Response MilliVolts 0 5 10 15 20 25 30 35 40 45 50 55 60 65 Time Minutes 64 0060 122 21 User manual Triathlon Peak rea number 206342 From the integration results the RSD for the Triathlon can 204773 be calculated 205425 205203 For this example the results 205566 205568 RSD 0 25 96 206156 206174 205956 The specified value is an RSD 196 206165 205765 205960 206296 205617 205619 205648 206147 207070 E 1 4 Mixing and carry over test The default system settings were used except for the following lt USAGE gt Use of mix methods ENABLED A tray with one sample vial filled with sample 50 ppm Uracil A01 one sample vial filled with eluent A02 and one empty destination A03 vial are placed in the Triathlon A 1 10 dilution of sample from the sample vial will be made in the empty destination vial After dilution from both vials sample and destination 10 uL will be injected The Reagent A vial is filled with eluent The wash solvent bottle is filled with 80 H2O 20 Iso propanol 0060 123 21 65 User manual Triathlon Inj
86. t vial appears to go to transport vials field lt TYPE_A gt to select type A tray segment 1 E to define position of the first transport vial 1 to define position of the last transport vial Escape Escape to return to the Ready Menu To program method Methods to enter the Methods Menu INJECTION 02 E to define method number 02 PICK UP 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 vial 10 E to define volume of 10 uL for 1st injection 10 E to define volume of 10 uL for 2nd injection 10 E to define volume of 10 uL for 3rd injection Escape to return to the Methods Menu WASH to enter the Wash Menu O1 E to define wash method number 01 INJECTION E to select wash between injections 300 to define the wash volume Escape Escape to return to the Ready Menu 0060 123 21 25 User manual Triathlon To define the series 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 02 E to define the location of the first sample vial 02 to define the location of the last sample vial Escape to return to Ready Menu To run the series Put vial with transport solvent mobile phase in transport vial position 1 and vial filled with sample at position 2 Make sure the transport vial is correctly
87. t will be used for analytical runs for explanation of air segment refer to Appendix H e headspace pressure to switch headspace pressure on or off The Triathlon uses headspace pressure to facilitate transport of sample into the loop The compressor will always be used during a wash procedure Please note that accuracy and reproducibility may decrease if headspace pressure is switched off However headspace pressure will only be useful if sample vials are airtight refer to sections 2 5 and 2 8 e time display offers a choice between two types of time representation e key click error beep and alarm buzzer offer the possibility to switch sound signals or off 34 0060 122 21 User manual Triathlon Table 5 1 Overview of General settings general setting for default possible range loop volume 100 uL 5 1000 ul needle tubing 15 ul 1 200 ul syringe volume 250 ul 100 250 500 1000 uL or 10 mL syringe speed normal low normal or high scale factor 1 0 0 1 1 0 needle height 2 0 40 skip missing vials yes yes or no air segment yes yes or no headspace pressure yes yes or no time base display HH MM SS H MM SS or H MM mm key click on on or off error beep on on or off alarm buzzer on on or off lt USAGE gt Press this key to enter the following Usage settings e protection code to enter a code for protection of methods Enter a six digit code 000000 999999 for protection of all methods Press CL to erase
88. the System Menu Tray Menu Tray segments can only be placed in an open position in the front half of the tray Select lt ADVANCE gt in the System Menu to rotate the tray if the Triathlon is not executing a run You can then place a tray segment in the front half of the tray Do not place or remove a tray segment at the rear half of the Triathlon tray as this may damage the tray sensors CAUTION 18 0060 122 21 User manual Triathlon Chapter 3 Getting started This chapter describes how to get started with the Triathlon The following will be explained e menus of the Triathlon e convenient working order e methods that can be programmed and linked to series e executing a series 3 1 Menus of the Triathlon The software of the Triathlon is menu driven The most important menus are e Ready Menu this menu appears after the Triathlon has been switched on It offers very general possibilities for among other things methods management and communication with other apparatus e System Menu this menu appears after the System key in the keypad has been selected It offers general possibilities for entering settings for an analytical run Please note that settings may have been factory installed to suit particular wishes of the user In that case no changes will have to be made in the System Menu You are advised to enable only those facilities that you will need to ensure that menus are as concise as possible e Methods Menu this me
89. the code 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 e timebase methods to enable or disable the possibility to program timebase methods The Triathlon controls other connected equipment during analysis time Program timebase methods in the Methods Menu Default disabled e mix methods to enable or disable the possibility to program mix methods for the Triathlon Program mix methods in the Methods Menu Please note that the Triathlon cannot analyze priority samples during a run if the mix method is enabled Default disabled e user program to enable or disable the possibility to program a user program If this function is enabled it is possible to enter a user program protection code 6 digits Program the user program in the Methods Menu Please note that the Triathlon cannot analyze priority samples during a run if the user program is enabled Default disabled e labeled vials to enable or disable the possibility to program labeled vials Program the location of labelled vials in the Series Menu Default disabled e templates to enable or disable the possibility to program templates Pogram templates in the Methods Menu Default disabled 0060 123 21 35 User manual Triathlon e calibration vials to enable or disable the possibility of programming calibration vials Program the location of the calibration vials in the Se
90. tical to those offered when the Triathlon is idle If a series or method is changed the new values become active the next time the Triathlon starts a series The series currently running are not affected by the changes 3 5 Executing a Series in remote control To execute a series from remote control execute the following steps 1 Press Start Stop 2 Enter the number of the first and the last series to be performed 3 Select lt REMOTE gt to enter the remote control mode The Triathlon will now operate as slave of another device and can be controlled with Next injection input and Next vial input To indicate that remote control is active an is displayed in the bottom left corner of the display during execution of the series At the end of the series the message Series completed via remote control is displayed 4 Press Escape to return to the Ready Menu Refer to Chapter 6 for more information on remote control User manual Triathlon 22 0060 122 21 User manual Triathlon Chapter 4 Using the Triathlon This chapter describes a number of examples of actions that can be performed with the Triathlon Please note that this chapter does not describe all types of actions that can be performed Try to do these examples to learn to work with the Triathlon These examples can be executed after the Triathlon has been installed in accordance with Appendix and after all items described in Chapter 2 have been correctly set
91. ticles in the sample and or mobile phase which can scratch the rotor seal Following is the procedure for changing the rotor seal and isolation seal Refer to figure F 2 and proceed Stator Stator Screws Figure F 2 Exploded view of model 7739 0060 123 21 69 User manual Triathlon Remove the three stator screws with the 9 16 hex key Remove the stator and stator ring from the valve body Pull the rotor seal off the pins Remove the isolation seal Mount the new isolation seal with the spring side facing away from the rotor seal Mount the new rotor seal The three pins on the shaft assembly fit into the mating holes in the rotor seal only one way Mount the seal with the grooves facing the stator 7 Replace the stator ring so that the body locating pin in the stator ring enters the mating hole in the body 8 Mount the stator on the valve so the stator locating pin in the stator ring enters the mating hole in the stator 9 Replace the three stator screws Tighten each an equal amount until the screws are tight D OR D F 2 Operating Instructions for Rheodyne Model 9740 Valve Shaft Assembly Shaft Assembly Rotated Rotated Clockwise Counterclockwise Figure F 3 Flow diagram of model 9740 as viewed from stator Model 9740 is a two position six port PEEK valve Figure F 3 shows the flow diagram of the valve The six small circles represent the ports in the valve stator The slots are the
92. to a series methods mix injection wash timebase can be linked to vials 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 vials in a series In this way all steps in an analytical run are laid down and stored 20 0060 122 21 User manual Triathlon e You can assign a user program to a series a possibility to combine all possible steps in the analytical process in one program The user determines the order of the separate actions the Triathlon has to perform 3 4 Executing a Series Execution of a series is only possible if you have programmed a method and defined a series for the samples you wish to analyse Series are not stored in battery backup and exist only for as long as the Triathlon is switched on Execute the following steps 1 Start programming the series by pressing Start Stop 2 Enter the number of the first series to perform and the number of the last series to perform 3 Select lt START gt to start the actual analytical run The Triathlon starts to execute the series you have defined After the Triathlon has executed the run the Ready Menu will appear again Refer to Chapter 4 for a number of examples illustrating this working order It is possible to program series and methods during run Press Series or Methods the possibilities offered in the menus are iden
93. to locate error appears on the display 0060 123 21 Check Appendix C message 81 User manual Triathlon Analytical problems In case of analytical problems you will have to determine whether they are caused by the autosampler or by the rest of the system Quick check Replace the valve by a manual injection valve to discriminate between valve problems and other problems Perform a number of manual Flushed loop injections If the results are good the autosampler is faulty if not the HPLC system should be checked Butter tubing Ve 0 Detector _ monomers Wash solvent ap 9 Sample In the flowcharts on the next pages it is assumed that the Triathlon does not display any error messages Please keep in mind that analytical problems might be caused by external influences like temperature and or light sensitive samples For this r
94. umber to enter lt NONE gt for wash method to define location of the first sample vial to define location of the last sample vial to return to the Ready Menu To run the series Place sample in position 1 of the tray Start Stop to start the Triathlon 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 Triathlon will now locate vial 01 and perform 10 UL partial loopfill injection The display of the Triathlon 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 vial on which the analysis is performed A 01 At the end of the defined analysis time the Ready Menu will be displayed again to indicate that the Triathlon is ready for the next analytical run 24 0060 122 21 User manual Triathlon Example 2 10 uL injection with uL pick up wash between injections In this example a different injection method is used than in the previous one For that reason several settings must be adapted in the System Menu For system settings System to enter the System Menu lt GENERAL gt E to enter the General Menu E until Air segment appears to go to the Air segment field lt NO gt E to switch off air segment Escape to return to the System Menu lt TRAY gt E to enter the Tray Menu E until transpor
95. xisting method The following types of steps can be programmed for a mix method lt ASPIRATE gt sample air destination reagent A D a programmed volume Speed of syringe can be selected from 1 9 Refer to table 5 1 page 34 for values Height H indicated is the distance of the needle point to the tray holder default 2 mm The maximum amount which can be aspirated is the total volume of the syringe lt DISPENSE gt sample waste destination reagent A D a programmed volume from the buffer tubing Speed of the syringe can be selected from 1 9 Refer to table 5 1 page 34 Height H indicated is the distance of the needle point to the sample tray 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 lt WAIT gt to define a pause H MM SS maximum of 9 hours 59 minutes and 59 seconds During the pause the needle will move to home position if the previous step is an aspirate or dispense action If you want the needle to stay in the same position an aspirate or dispense step of 0 uL must be programmed at the desired position lt REPEAT gt Enter the number of steps that must be repeated and how often they must be repeated lt WASH gt enter the volume for needle wash Buffer is rinsed to waste 38 0060 122 21 User manual Triathlon
96. yringe speed Use user program 4 type number Input edge next inj Needle height User program protection 5 type number Input edge next vial Skip missing vials code 6 type number Freeze input active Air segment Use labeled vials 7 type number Reset outputs after Headspace pressure Use templates 8 type number series Time base display Use calibration vials number of transport vials Key click Error beep Alarm buzzer Methods programming Method lt TEMPLATE gt lt METHODE gt lt USRPROG gt Userprog User program or ASPIRATE Mix method Methods DISPENSE Injection method lt INJECTION gt lt WASH gt lt TIMEBASE gt lt MIX gt SYR VALVE Wash method SYRINGE Teese Injection methods e Wash volume Timebase program Mix method bp lt FLUSHED gt lt PARTIAL gt lt PICK UP gt e Wash between 4x AUX 1 e ASPIRATE WAIT Series e 4x AUX 2 sample COMPRESS Analysis time Vials 4x AUX 3 destination Injections e 4x AUXA air Ax ISSA ad e Ax SS B DISPENSE PROG OUT e Inj volumes CODE e 8x SSV sample 8x CODE OUT END TIME Pen reagent REPEAT WAIT WASH With templates Without templates Series e Template Userprogram yes no e Use calibration vials yes no or calibration vials Mix method calibration interval Injection method e Sample vials Wash method e Destination vials Timebase method s Reagent vials Use calibration vials yes no Note e Labeled vials c

user manual

Contents

Download Pdf Manuals

Related Search

Related Contents