MassLynx NT Guide to Inlet Control

Transcription

1 MassLyx NT Guide to Ilet Cotrol Versio rd August 2000 Page i

2 MassLyx NT Guide to Ilet Cotrol The software described i this book is furished uder a licece agreemet ad may be used oly i accordace with the terms of that agreemet. Copyright Notice Micromass Ltd believes that the iformatio i this publicatio is accurate. However the iformatio is subject to chage without otice ad should ot be costrued as a cotractual udertakig by Micromass Ltd. Despite the care which has bee give to the preparatio of this publicatio, Micromass Ltd accepts o resposibility for ay loss or ay other matter which may arise from ay error or iaccuracy which may iadvertetly have bee icluded. Copyright (C) Micromass Ltd. All Rights Reserved. No part of this publicatio may be copied without the express writte permissio of Micromass Ltd. Trademarks Micromass is a registered trade mark of Micromass Limited (Reg. U.S. Pat. & Tm. Off.). MassLyx is a registered trademark of Micromass Ltd. Widows is a trademark of Microsoft Corporatio. Other product ames metioed i this maual may be trademarks or registered trademarks of their respective compaies ad are hereby ackowledged. Page ii

3 Table of Cotets Cotrollig Ilet Systems ad Autosamplers 1 Itroductio 1 The Ilet Editor 2 The Ilet Editor Toolbar 3 The System Status Page 4 Ilet Cofiguratio 6 Waters Systems 11 Waters 600 Pump 11 Waters 2690 Autosampler 15 Waters 2690 Pump 17 Waters 996 PDA Detector 23 Waters 486 UV Detector 26 Waters 2487 UV Detector Sigle Wavelegth Absorbace Detector Dual Wavelegth Absorbace Detector 27 Waters 2487 IEEE Detector 28 Waters SAT/IN PDA Detector 29 Waters 2700 Autosampler 30 Waters 2700 Bed Layout 33 Waters 2700 Fixed Positios 36 Waters 2700 Plate Geerator 37 Waters 2700 Meu 39 Waters 2790 Autosampler 40 Waters 2790 Pump 44 Waters 2790 Meu 53 Waters 2790 Plate Geerator 55 Waters 2790 Bed Layout 58 Waters CapLC System Status Pages 60 Waters CapLC Pump 63 Waters CapLC Autosampler 71 Waters CapLC Bed Layout 74 Waters CapLC Plate Geerator 77 Waters CapLC Plate Loader 80 Waters CapLC PDA Detector 81 Waters 515 ad 1525 Pumps 84 CE Istrumets 91 CE Istrumets GC8000 Gas Chromatograph 91 CE Istrumets AS800 Auto Ijector 94 Gilso Systems 97 Gilso Autosamplers 97 Itroductio 97 The Gilso Toolbar 98 Gilso Cofiguratio Pages 98 Gilso Advaced Optios 108 Gilso Pump 118 Pump Cofiguratio 121 Page iii

4 Hewlett Packard Systems 125 Hewlett Packard 5890 Gas Chromatograph 125 Hewlett Packard 7673A Auto Ijector 127 Hewlett Packard HPLC Systems 128 The HP6890 System Status Page 137 The HP6890 Toolbar 153 Jasco Systems 157 Jasco 900 ad Jasco 1500 Autosamplers 157 Jasco 900 ad Jasco 1500 Pumps 158 Jasco 900 ad Jasco 1500 UV Detectors 160 Shimadzu Systems 163 Shimadzu Systems 163 Shimadzu Pump 165 CTC, Cetac ad Other Systems 171 CTC A200S Autosampler 171 CTC PAL Autosampler 174 PAL Cycle Composer Method Editor 175 Usig the PAL CTC Autosampler with OpeLyx179 Cetac ASX100 Autosampler 181 Cetac ASX500 Autosampler 182 Solids Probe 183 DCI Probe 184 Thermospray Probe 184 RoboProbe 184 Cotact Closure 185 Idex 189 Page iv

5 Cotrollig Ilet Systems ad Autosamplers Itroductio Chapter 1 Ilets Mass spectrometers are usually used i cojuctio with a ilet system such as a liquid chromatograph (LC) or a gas chromatograph (GC). MassLyx ca cotrol this equipmet durig data acquisitio to provide complete cotrol of a experimet. Autosamplers will ofte be used to automate the ruig of samples. Acquisitios that use a ilet system ca oly be started from the Istrumet Cotrol Pael ad caot be started from the tue page. Before you try to use a ilet system for the first time you must cofigure MassLyx to use that ilet. To select a ew ilet system, choose Select Iterface from the Acquisitio Cotrol Pael Cofigure meu. The list of ilet optios that appears i the Select Iterface dialog reflects the ilet systems, which were selected whe MassLyx was istalled. For some Ilet systems (HP1090, HP1050, HP1100, HP6890, Jasco, Waters, MicroTech ad Gilso) selectig the GC or LC system (ACE) (Aalytical Compoet Egie) optio allows the user to chage the Ilet, Autosampler ad Detector without havig to re-istall MassLyx. For other o ACE systems if, at a later date you add a ew ilet system or chage oe of your existig ilets you may eed to re-istall MassLyx to gai access to the cotrol software for the ew ilet system. The method used to cotrol the ilet system is set up before you start to acquire ay data ad is saved o disk for use by the acquisitio system. Differet methods ca be saved, accessible by ame i the usual maer. You must supply the ame of the ilet method that you wish to use whe you start a acquisitio by eterig it ito the Ilet field i either the Sigle or Multiple sample start editors (these are covered i the ext sectio). N.B. Make sure that ay chages that you make to a ilet program are saved to disk before you start a acquisitio. This is doe by selectig the Save optio o the File meu of the ilet editor or by pressig the toolbar butto. If you do ot save the parameters the the previous oes will be used as MassLyx reads the parameters from disk, ot from the editor, whe it starts to acquire. Icoisig the display does ot save the parameters but you will be give the optio to save ay chages that you have made if you actually close the editor. Autosamplers A autosampler ca oly be used with the multiple sample acquisitio editor o the MassLyx top level scree. The rules regardig the savig of parameters for ilet editors apply to autosampler editors as well. Page 1

6 The Ilet Editor The Ilet Editor is used to View the status of the curret system. Defie the GC or LC, autosampler ad detector methods. Chage istrumet cofiguratio Cotrol pumps ad lamps ad ru methods. To access the Ilet Editor press the toolbar butto o the LC pael of the MassLyx top level scree, or select Ilet from the Methods meu o the Acquisitio Cotrol Pael. Figure 1.1 Ilet Editor dialog System Status page Page 2

7 The Ilet Editor Toolbar The toolbar is displayed across the top of the applicatio widow, below the meu bar. The toolbar provides quick mouse access to may tools used i the cotrol software. Click To Create a New method. Ope a existig method. Save the method with its curret ame. Prit the curret method. Display the System Status dialog. Edit Ilet system parameters. Edit AutoSampler parameters. Edit Detector parameters. Start or stop the pump. Tur Lamp o ad off. Ru the curretly saved method Load the curretly saved method. Display the Help Cotets. Page 3

8 The System Status Page The System Status page displays iformatio about the state of the machie beig cotrolled. This page ca be accessed withi the Ilet Editor by selectig Status from the View meu or by pressig the toolbar butto. Note: This chages for a GC, see HP6890 later i the maual. The Waters Cap LC also has a differet System Status page see the Waters Cap LC System Status Page later i this chapter. Figure 1.2 System Status page Idicators The Ruig, Pump O ad Ijector Cycle idicators at the left-had side of the scree give you iformatio o the curret status of the LC system. The OK ad Ready Idicators become illumiated i red if the LC System has a error. You ca the click o the red idicators to give you more iformatio o the cause of the malfuctio. Ru Time Displays how log the method has bee ruig. Flow Rate This is the curret flow rate as retured by the istrumet. Pressure Displays the curret pressure i the istrumet. Colum Left ad Colum Right Displays the curret temperature of the left ad right colums. These will be grayed out if colum heaters are ot istalled. Sample Temp Displays the curret temperature of the sample. This will be grayed out if a sample heater is ot istalled. Gradiet Displays the solvet percetages at which the LC System is curretly operatig. PDA Detector Whe acquirig diode array data the Diode Array Status displays the umber of scas curretly acquired. Page 4

9 Savig ad loadig LC parameter files The Curret LC parameters ca be saved to disk by choosig Save or Save As from the Ilet Editor File meu. A set of previously saved LC parameters ca be recalled from disk by choosig Ope from the Ilet Editor File meu. To prit a LC Method Report Choose Prit from the Ilet Editor File meu or press the toolbar butto. Press OK to prit a report detailig the parameters used i the curret LC Method. To Dowload parameters to the LC system To dowload the parameters to the LC system, press the Method from the LC meu. butto or choose Load The status bar will idicate the progress of dowloadig the parameters. Oce values have bee dowloaded you ca start the pump ruig with the iitial coditios. To Ru the Pump with iitial coditios Select Pump O from the LC meu or click o the begi ruig with its iitial coditios. butto. The pump will To tur o the Lamp Select Lamp O from the LC meu or click o the butto. To begi a Gradiet Method or start a ijectio You ca ru a sigle ijectio with the Autosampler by selectig Ru Method from the LC meu as soo as the meu item is eabled ( it is disabled whe the system is ruig a method ). If a method is already ruig i the LC System it will ot be possible to start a ew method ( either iject or ru gradiet oly ) util the previous method has stopped. Selectig Ru Method (No Ijectio) from the LC meu starts the gradiet (if etered) to allow maual ijectios. Page 5

10 Ilet Cofiguratio As metioed earlier selectig the GC or LC system (ACE) optio, from the Select Iterface dialog, allows the user to chage the Ilet, Autosampler ad Detector without havig to re-istall MassLyx. The Ilet Cofiguratio dialog gives a list of the Ilet systems, Autosamplers ad detectors supported by MassLyx. To access the dialog select Istrumet Cofiguratio from the Tools meu o the Ilet Editor. Istrumets Figure 1.3 Istrumet Cofiguratio dialog Check the boxes for the required cofiguratio ad press OK. MassLyx will iform the user if the cofiguratio is ot supported. Page 6

11 HPIB Commuicatio Figure 1.4 HPIB Commuicatio dialog Whe a Istrumet cofiguratio has bee selected o the Istrumet tab default device addresses are writte to this dialog. Values ca be chaged if required. For the HP1100 DAD detector the PC Coectio should be set to HP100 DAD. GPIB Commuicatio Figure 1.5 GPIB Commuicatio dialog Whe a Istrumet cofiguratio has bee selected o the Istrumet tab default IDs are writte to this dialog. These may eed chagig ad will be defied o setup by a Micromass egieer. For more iformatio cosult the relevat istrumet istructio maual. Page 7

12 Page 8

13 Notes Page 9

14 Notes Page 10

15 Waters Systems Waters 600 Pump Chapter 2 The Waters 600 Pump pages ca be accessed by selectig Waters600 Pump from the View meu o the Ilet Editor or by pressig the toolbar butto. Waters 600 Iitial Coditios Figure 2.1 Iitial Coditios page Solvets Up to four solvets will be displayed depedig upo the system cofiguratio. The total value of all the solvet percetages added together must ot exceed 100%. Pump A This is the remaider percetage after the solvet percetages have bee set for the other pumps. Pump B, C ad D These ca either be eabled or disabled by checkig the box ext to the idividual pumps. The values ca be set to the required percetage of flow delivery. Solvet Name Eter the ame of the solvet that will be delivered through the correspodig Pump. Flow Rate This is the total flow rate of the solvet chaels accordig to how you have cofigured the istrumet. Pressures Eter the upper ad lower limits of the pressure withi the solvet delivery system (SDS) if the pressure falls outside of this rage the SDS switches off. Page 11

16 Colum Heater If the istrumet has a ove preset the the colum temperature ca be set to a specified temperature i degrees cetigrade. Eter the temperature to heat the colum to i the Set box ad a High Limit. If the temperature exceeds the High Limit the the system will shut dow. If the software has bee cofigured to operate without a colum ove the these boxes will be greyed out. Ru Time Eter the time i miutes that the method will ru from the poit of ijectio. Waters 600 Gradiet Page Figure 2.2 Gradiet page This page allows a gradiet to be etered ad edited. To operate i isocratic mode esure that the timetable is empty. To eable the B%, C% ad/or D% boxes check the relevat boxes o the Iitial Coditios page. To add a gradiet, eter a time ad percetage i the relevat boxes ad press the toolbar butto. Note: The first etry must have a time of 0. To delete a sigle gradiet, click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To modify a gradiet, select the required etry i the timetable. The values will the be displayed i the edit boxes to the left of the timetable, ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig timetable. will result i a ew etry beig created i the Page 12

17 Flow Eter the flow rate for the solvet delivery system. Curve This sets the rate at which the solvet is to chage to the ew proportios ad/or flow rates. See the Waters 600 Operator s Guide for a list of values. Waters 600 Iitial Evets Page Figure 2.3 Iitial Evets page This page allows the iitial state of switches 1 to 4 to be defied. Check the box(es) for the switches that should have a iitial state of off. Page 13

18 Waters 600 Programmed Evets Page Figure 2.4 Programmed Evets page This page allows pump evets to be etered ad edited. To add a evet, type i a time, select a evet from the drop dow list box, select a actio or eter a value ad press the toolbar butto. To delete a sigle evet, click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To modify a evet, select the required etry i the timetable. The values will the be displayed i the edit boxes above the timetable, ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig will result i a ew etry beig created i the timetable. Page 14

19 Waters 2690 Autosampler Note: To cotrol the Waters 2690 autosampler ad pump from the keypad rather tha the MassLyx software the Ilet must be cofigured as Noe. See Cofigurig the Ilet System i the Acquisitio Cotrol Pael chapter. Waters 2690 Autosampler Iitial Coditios Page This page is used to set parameters specific to the Sampler, to access it select Waters2690 AutoSampler from the View meu or press the toolbar butto. Figure 2.5 Autosampler Iitial Coditios page Sample Heater Temperature If the sample heater is istalled, eter the temperature that the sample should be to be heated or cooled to. Rage: 4.0 to 40.0 C. Sample Heater Temperature Limit Eter the maximum deviatio i sample temperature allowed. If this is exceeded the curret acquisitio will stop ad the LC Status error light, o the MassLyx scree, will tur red. Rage: ±1.0 to ±20.0 C. Ijectio Volume Eter the volume of sample to be ijected, i microlitres. Rage: 0 to 2000 µl. Note: If you are ruig from the Sample List, the ijectio volume i the sample list etry overrides the value etered here. Needle Depth This adjusts the depth of the eedle tip to accommodate for sedimeted samples or o-stadard vials. A value of 0 correspods to the bottom of the vial. Rage: 0.0 to 20.0 mm i 0.1mm icremets. Page 15

20 Draw Speed This determies the rate i microlitres per secod at which sample is extracted ito the autosampler eedle. It should be set accordig to the viscosity of the sample. Select oe of Fast, Normal or Slow from the dropdow list box. The table below shows the draw rate for each selectio usig a 250 µl syrige. Selectio Fast Normal Slow Draw Rate for a 250 µl Syrige 5.0 µl/sec 2.5 µl/sec 1.0 µl/sec Optioal Sample Loop To iject sample volumes greater tha 100 microlitres a additioal sample loop ca be istalled (i series with the existig sample loop), check this box if a additioal sample loop is used. Vial umber The vial to iject from. Note: If a multisample acquisitio is beig ru from the MassLyx Sample List, the Bottle # etry i the sample list overrides the value etered here. Waters 2690 Autosampler Purge Page This page is used to set the Autosampler purge volume, to access it click o the Autosampler Purge tab. Figure 2.6 Autosampler Purge page Loop Volumes Eter the umber of times the loop should be filled to purge the sample loop ad syrige of traces of the previous sample. Whe set to a value greater tha zero, this actio is performed after every ijectio. Page 16

21 Waters 2690 Pump Note: To cotrol the Waters 2690 autosampler ad pump from the keypad rather tha the MassLyx software the Ilet must be cofigured as Noe. See Cofigurig the Ilet System i the Acquisitio Cotrol Pael chapter. The Waters Pump pages ca be accessed by selectig Waters2690 Pump from the View meu o the Ilet Editor or by pressig the toolbar butto. Waters 2690 Solvets ad Flows Page Figure 2.7 Solvets ad Flows page Solvets Up to four solvets will be displayed depedig upo the system cofiguratio. The total value of all the solvet percetages added together must equal 100%. Solvet Names etered here will be displayed o the Pump Gradiet page. Pump A This displays the remaider percetage after the solvet percetages have bee set for the other eabled pumps. Pump B, C ad D These ca either be eabled or disabled by checkig the box ext to the idividual pumps. The values ca be set to the required percetage of flow delivery. Ru Time This value is set to the time i miutes that the method will ru from the poit of ijectio. Degasser This value is set to the time i miutes that the method will ru from the poit of ijectio. Flow This is the total flow rate for the system. Flow Ramp Eter the time (i miutes) for the solvet delivery system to reach the maximum system flow rate (10 ml/mi). Recommeded miimum settig: 0.5 mi. Page 17

22 Waters 2690 Colum Setup Page Figure 2.8 Colum Setup page Colum Heater Temperature Eter the target operatig temperature for the optioal colum heater. This value must be at least 5 C above ambiet. Rage: 20 to 60 C. Colum Heater Temperature Limit This is the maximum deviatio i colum temperature allowed. If this is exceeded the curret acquisitio will stop ad the LC Status error light, o the MassLyx scree, will tur red. Rage: ±1 to ±20 C. Eter Low Pressure ad High Pressure values as required. If the pressure falls outside these limits the curret acquisitio will stop ad the LC Status error light, o the MassLyx scree, will tur red. Low Pressure Rage: 0 to 310 bar. Low Pressure Rage: 0 to 345 bar. Pre-colum Volume Eter the volume of solvet to pump through the colum before a ijectio. Rage: 1 to µl. Page 18

23 Waters 2690 Pump Gradiet Page Figure 2.9 Pump Gradiet page This page allows a gradiet to be etered ad edited. If you wish to operate i isocratic mode the you should eter parameters o the Solvets ad Flows page ad esure that the timetable is empty. Time (mis) Specifies the time at which the specified coditios (%A to %D, Flow, ad Curve) for the row should take effect. Make sure you set Time for the first row to 0.00, to establish iitial coditios for the gradiet ru. The rage for rows other tha row 1: 0.01 to miutes. Solvet A % Solvet D% Specifies the percetage of solvet flow from each reservoir. For each row, the total of all solvets must equal 100%. Rage: 0 to 100%. Flow (ml/mi) Specifies the total flow rate for the solvet delivery system. Rage: 1 to 10 ml/mi. Curve This sets the rate at which the solvet is to chage to the ew proportios ad/or flow rates. Curves are specified by umber. Available choices: 1 to 11. Curve Number Effect 1 Immediately goes to specified coditios 2 to 5 Covex 6 Liear 7 to 10 Cocave 11 Maitais start coditio util ext step Page 19

24 Curve Profiles Waters 2690 Gradiet Table Operatio To add a gradiet, eter values i the relevat boxes ad press the toolbar butto. You ca add up to 15 rows to the table. Note: The first etry must have a time of 0. To delete a sigle gradiet, click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To modify a gradiet, select the required etry i the timetable. The values will the be displayed i the edit boxes above the timetable, ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig will result i a ew etry beig created i the timetable. Page 20

25 Waters 2690 Pump Evets Page Figure 2.10 Pump Evets page This page allows pump evets to be etered ad edited. Use the Evet Table to program up to 16 evets (both exteral ad iteral). The exteral evets are triggered by four cotact closures (relays) through output termials (S1 S4) o the 2690 Separatios Module. The iteral evets are used to cotrol the sample compartmet temperature ad colum heater temperature, ad to prime ad flush the 2690 Separatios Module. Evets ca be triggered more tha oce ad multiple evets ca be triggered simultaeously. Time Eter the time at which the evet should start. Evet rows are sorted automatically by time. Note: Differet evets ca be programmed to occur at the same time. Rage: 0.00 to mi. Evet Eter the type of evet sigal required: oe of the four TTL-level output switches (S1 S4), or oe of the iteral evets (colum heater temperature, sample compartmet temperature or flush/prime). Available choices: Switch 1 to Switch 4 Correspods to termial strip positios S1 to S4 o the rear of the 2690 module. Activatig a Switch evet triggers a cotact closure for cotrollig a exteral device. After selectig a switch evet, set a state for the switch by selectig O, Off, Toggle, Pulse Width or No Chage. This state appears i the Actio colum of the table (refer to Switch States, below). Note: If Pulse is selected for a switch state, the duratio of the pulse must be etered i the Width (mi) field. Set Temperature (Colum or Sample) Specifies the temperature of a optioal colum heater, or a optioal sample compartmet heater/cooler. After selectig this evet, select Colum or Sample ad eter the required Temperature i C. Note: Whe a Colum Temperature evet occurs, the temperature of the colum heater chages from the value set i the Heaters ad Pressures page to the value set for the evet. Whe the evet times out, the temperature chages back to the Heaters ad Pressures page value. Colum rage: 20 to 60 C. Sample rage: 4 to 40 C. Page 21

26 Flush/Prime Specifies a flush/prime operatio for the 2690 module. Use this evet oly whe creatig Ilet Pre-ru ad Ilet Post-ru methods. These methods will use the solvet percetages ad the ru time from the Solvets ad Flows page but will use the Flow value etered o this page. Note: The Time field is ot accessible whe you select a Flush/Prime evet. Switch States O Turs o a cotact closure that triggers a exteral or iteral evet. With this fuctio, the cotact closure remais closed util a Off fuctio is set. Off Turs off the cotact closure for the evet. With this fuctio, the cotact closure is broke. Pulse Trasmits a sigle O/Off pulse. The cotact closure is maitaied for the umber of secods defied i the Value colum. Rage: 0.01 to sec. Toggle Chages the curret state of the switch. No Chage Leaves the switch i its curret state. Waters 2690 Evet Table Operatio To add a evet, eter a time, select a evet from the drop dow list box, select a actio ad press the toolbar butto. Up to 16 evets ca be programmed. To delete a sigle evet, click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To modify a evet, select the required etry i the timetable. The values will the be displayed i the edit boxes above the timetable, ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig will result i a ew etry beig created i the timetable. Page 22

27 Waters 996 PDA Detector This page is used to set parameters specific to the UV detector, to access it select Waters996 PDA Detector from the View meu or press the toolbar butto. Waters 996 PDA Page Figure UV Detector Cofiguratio page Start Wavelegth Eter the wavelegth at which to start acquirig data. Ed Wavelegth Eter the wavelegth at which to stop acquirig data. The rage with Resolutio set to 1.2 is m to m. The rage at all other Resolutio settigs is (Resolutio/2) to (Resolutio/2). Resolutio Eter the umber of diodes that are averaged together as a sigle spectral data poit. To differetiate closely related spectra ad obtai greater spectral resolutio, use a small resolutio umber. Be aware, however, that a small resolutio value geerates more data poits ad therefore requires more disk space tha a large resolutio value. Fid a resolutio value just small eough to idetify spectral features. Rage: 1.2 to 24.0 m i multiples of 1.2. Samplig Rate Select the umber of Spectra to be acquired per secod, from the dropdow list box. For good itegratio ad quatitatio, acquire 15 to 20 spectra across a peak. Auto Exposure Check this box to eable the detector optics to calculate the optimum exposure time eeded to recharge the diodes, based o the lamp eergy, the lamp spectrum ad the selected wavelegth rage. Tip: Eable Auto Exposure for most routie aalyses. Iterpolate Check this box to istruct the detector to igore the sigal from the photodiode at 656 m ad iterpolate a value from the adjacet diodes. This prevets over-saturatio at 656 m (Balmer lie for deuterium). Oly applicable if the Auto Exposure optio has bee selected. Page 23

28 If this box is ot checked the detector reports the sigal from the photodiode at 656 m, this is oly ecessary if you are workig with compouds that absorb i the 656 m rage. Note: If this parameter is uchecked, the deuterium lamp high emissio lie at 656 m may cause spectral artifacts ad autoexposure errors. Exposure Time The exposure time is the time that the photodiodes are exposed to light before they are read. To set a differet Exposure Time, esure that the Auto Exposure box is ot checked ad eter the required time i milli secods. Rage: to ms. Stop Time To specify a differet Acquisitio Stop Time eter the time i miutes whe the PDA should stop scaig. Filter Respose Eter the respose time for filterig acquired data. The filter is a ehaced rollig average filter applied to absorbace data from the PDA detector before the data is set to MassLyx. The filter reduces high-frequecy oise across the etire wavelegth rage specified for the acquisitio. High values decrease peak respose. Available choices: 0, 1, 2 ad 3. Save to Disk Check this box to save the Photo Diode Array data to the raw datafile. If this data is ot required for further processig the ucheck the box, the data is ot saved to disk thus reducig the size of the file. Waters 996 Chael Detector Cofiguratio Pages The Chael 1 ad Chael 2 pages cotai the same iformatio. Select the page relevat to the chael required, by clickig o the tab. Figure 2.12 Chael 1 Detector Cofiguratio page Page 24

29 Output Mode Select oe of Off o aalog output sigal. Absorbace Output is i absorbace uits at the wavelegth specified. Note: Ratio Deomiator Wavelegth ad Threshold parameters are ot accessible whe Absorbace mode is selected. Ratio Output represets the ratio of absorbaces at two wavelegths. The umerator wavelegth is specified by the Wavelegth parameter, ad the deomiator wavelegth is specified by the Ratio Deomiator Wavelegth parameter (see below). Wavelegth Eter the output wavelegth to moitor. I Ratio mode, the absorbace at the Wavelegth is used to calculate ratio i the formula: Ratio = Absorbace at Wavelegth/Absorbace at Ratio Deomiator Wavelegth Wavelegth must be withi the wavelegth rage specified by the Start Wavelegth ad Ed Wavelegth parameters o the PDA page. Rage whe Resolutio is set to 1.2: Start Wavelegth to Ed Wavelegth. Rage at all other Resolutio settigs: Start Wavelegth + (Badwidth/2) to Ed Wavelegth (Badwidth/2). Default: 254 m. Ratio Deomiator Wavelegth Eter the deomiator wavelegth (i aometers) for the aalog output chael. Ratio Deomiator Wavelegth must be withi the wavelegth rage specified by the Start Wavelegth ad Ed Wavelegth parameters o the 996 PDA page. Badwidth Eter the spectral badwidth of the aalog output chael. The rage is 1.2 to 24.0 m. Filter Type Select Hammig or Sigle Pole from the dropdow list box. The Hammig filter is desiged to create the same degree of peak-height degradatio as the Sigle Pole filter for the same respose time, but ehaces filterig of high-frequecy oise. Filter Respose Eter the respose time for the filter. The rage is 0 to 5 secods. Offset If required eter a offset to the aalog output chael. The rage is 0.2 to 2.0 AU. Threshold Eter a threshold above which the ratio (Wavelegth / Ratio Deomiator Wavelegth) must be to be valid data. The rage is 0.1 to 2.0 AU. Note: If o ratio is plotted oe or both chaels are below the curret Threshold ad a lower Threshold value should be etered. Page 25

30 Waters 486 UV Detector This page is used to set parameters specific to the UV detector, to access it select Waters486 UV Detector from the View meu or press the toolbar butto. Figure UV Detector Cofiguratio page Wavelegth (µ) Eter the wavelegth to moitor. Polarity Select the polarity of the output sigal from the drop dow list box. Sesitivity (AUFS) Eter the required sesitivity of the output sigal. Time Filter (secods) Eter the respose time for filterig acquired data. A full descriptio of all the parameters i this editor is give i the Waters 486 Istructio Maual. Page 26

31 Waters 2487 UV Detector This page is used to set parameters specific to the Waters 2487 UV detector, to access it select Waters2487 UV Detector from the View meu or press the toolbar butto Sigle Wavelegth Absorbace Detector The 2487 detector ca be used as a sigle or dual wavelegth detector. To use as a sigle wavelegth detector select Sigle Wavelegth from the Waters2487UV meu. A tick mark will appear ext to the ame if sigle wavelegth is selected ad the 2487 Chael B parameters are greyed out. Figure UV Detector Cofiguratio page (Sigle Wavelegth) Wavelegth (µ) Eter the wavelegth to moitor. Sesitivity (AUFS) Eter the required sesitivity of the output sigal. Time Filter (secods) Eter the respose time for filterig acquired data. Polarity Select the polarity of the output sigal from the drop dow list box. A full descriptio of all the parameters i this editor is give i the Waters 2487 Istructio Maual Dual Wavelegth Absorbace Detector The 2487 detector ca be used as a sigle or dual wavelegth detector. To use as a dual wavelegth detector esure that the Sigle Wavelegth optio o the Waters2487UV meu is ot selected. If a tick mark appears ext to the ame the sigle wavelegth is selected, selectig the optio agai will retur the detector to dual wavelegth mode ad both chael parameters will be available. The parameters are the same as for sigle wavelegth mode. Page 27

32 Waters 2487 IEEE Detector The 2487 IEEE Detector should be selected if the detector is coected via a GPIB card i the back of the PC. The 2487 IEEE Detector is used to collect biary data, through the IEEE iterface, rather tha through the aalog iterface. This page is used to set parameters specific to the Waters 2487 IEEE detector, to access it select Waters2487 IEEE Detector from the View meu or press the toolbar butto Dual Absorbace Detector The 2487 detector ca be used as a sigle or dual wavelegth detector. To use as a sigle wavelegth detector select Sigle Wavelegth from the Waters2487UV meu. A tick mark will appear ext to the ame if sigle wavelegth is selected ad the 2487 Chael B parameters are greyed out. Figure UV Detector Cofiguratio page (Sigle Wavelegth) Wavelegth (µ) Eter the wavelegth to moitor. Sesitivity (AUFS) Eter the required sesitivity of the output sigal. Time Filter (secods) Eter the respose time for filterig acquired data. Polarity Select the polarity of the output sigal from the drop dow list box. A full descriptio of all the parameters i this editor is give i the Waters 2487 Istructio Maual. Page 28

33 Waters SAT/IN PDA Detector This page is used to set parameters specific to the PDA detector, to access it select WatersSATIN PDA Detector from the View meu or press the toolbar butto. Figure 2.16 SAT/IN Cofiguratio page Chael 1 ad Chael 2 Check the box(es) for the required chaels. Samplig Rate Select the umber of Spectra to be acquired per secod, from the dropdow list box. For good itegratio ad quatitatio, acquire 15 to 20 spectra across a peak. Notes Data collected through a Waters SAT/IN PDA Detector is show as aalog data i the acquired data files, this is the same as aalog data comig i through the aalog iputs from the back of a MS istrumet. The Waters SAT/IN PDA Detector ca be used to collect aalog data or the MS aalog iputs ca be used. Do ot try to collect aalog data with the SAT/IN ad the MS aalog iputs at the same time. Collectig aalog data from both sources will result i upredictable behaviour. SAT/IN aalog data will oly be collected i a system cofigured with a spectral data source. A MS detector ad/or a PDA detector must also be used to successfully collect SAT/IN data. Negative data is ot supported by the Waters SAT/IN but ca be avoided by applyig a appropriate offset i the coected Detector. Page 29

34 Waters 2700 Autosampler These pages are used to set parameters specific to the autosampler, to access them select Waters2700 AutoSampler from the View meu or press the butto. toolbar Waters 2700 Ijectio Cofiguratio Figure 2.17 Waters 2700 Sampler Ijectio Cofiguratio Page Loop Volume (µl) Eter the volume of the sample loop i microlitres. Ijectio Volume (µl) Eter the volume of the sample to iject ito the loop for sigle sample acquisitios. For samples acquired via a sample list this is overridde by the value i the sample list. If the Ijectio Volume is equal to the Loop Volume the twice the Ijectio Volume is draw to esure that the loop is full. Separatio Air Gap (µl) Eter the volume of air to draw before the sample. Vial Eter the positio of the vial to use for sigle sample acquisitios. For samples acquired via a sample list this is overridde by the value i the sample list. Page 30

35 Waters 2700 Dilutor Cofiguratio Figure 2.18 Waters 2700 Sampler Dilutor Cofiguratio Page Syrige Size Select the size of the syrige istalled from the drop dow list box. Aspiratio Speed Eter a value for the speed at which to draw the sample ito the eedle (the pump will be o its dowward jourey). Rage: 1 to 32, with 1 beig the fastest. Dispese Speed Eter a value for the speed at which to eject the sample from the eedle (the pump will be o its upward jourey). Rage: 1 to 32, with 1 beig the fastest. Waters 2700 Wash Parameters Figure 2.19 Waters 2700 Sampler Wash Parameters Cofiguratio Page Page 31

36 Needle Rise Volume (µl) Eter the volume of mobile phase required to wash the eedle after a ijectio. A value of zero will result i o wash. If the eedle rise volume is greater tha 800µl the the mii-wash pump is used istead. Wash Time (secods) Eter the time for which the mii-wash pump is activated durig a mii-wash prime. Mii-wash prime is activated from the Waters2700 meu. Ijectio Port Flush Volume (µl) Eter the volume of mobile phase required to flush the iject port after the sample has bee ijected. A value of zero will result i o port flush. Ijectio Port Flush Speed Eter the speed at which the flush volume is dispesed. Rage: 1 to 32, with 1 beig the fastest. Waters 2700 Sample Cofiguratio Figure 2.20 Waters 2700 Sampler Cofiguratio Page Curret Layout This shows the curretly selected rack cofiguratio. To chage the curret layout select a ew oe from the drop dow list box. Page 32

37 Waters 2700 Bed Layout Bed layouts are created, deleted or ameded from this dialog. To display the Bed Layout Editor dialog, select Bed Layout from the Waters2700 meu. Figure 2.21 Bed Layout Dialog To Create A New Bed Layout (Waters 2700) 1. Highlight a bed layout similar to the oe you wat to create ad press the butto to create a ew layout. The layout appears i the Bed Layouts list as the same ame with a 1 at the ed, e.g. Six Positio Microtiter1. 2. To chage the ame of the layout, type the ew ame ito the Bed Layouts text box ad press the list box. butto. The ame is updated i the Bed Layouts New bed layouts are saved to the MassLyx Racks directory. To Delete A Bed Layout (Waters 2700) 1. Highlight the bed layout to delete ad press the butto. A dialog box will ask you to cofirm the deletio. Press the OK butto to delete the bed layout. Note: The bed layout which is selected as the curret bed layout o the Sampler Cofiguratio page caot be deleted. Page 33

38 Other Bed Layout Optios (Waters 2700) 1. To chage the umber of rows i the curret colum, type the ew umber ito the Rows box ad press the butto. 2. To apped a ew colum, press the butto. 3. To delete the curret colum press the butto. 4. To isert a colum, click o the colum before which you wat to isert ad press the butto. Note: The colum iserted will have the same umber of rows as the colum highlighted. Modify Bed Layout (Waters 2700) If the plate positio or type eeds chagig select the Modify Bed Layout tab. Figure 2.22 Modify Bed Layout Dialog Click o oe of the code plates to display the Plate Positio ad Type dialog. Page 34

39 Figure 2.23 Plate Positio ad Type dialog This dialog allows you to select a ew plate from a list of possible optios, ad chage its actual positio o the bed. Measuremets for plate positios are always take from the top left corer of each plate. The X value is the measuremet from the vial positio i the top left corer of the plate to the home positio. The Y value is the measuremet from the vial positio i the top left corer of the plate to the home positio. Vial bottom from probe home This is the distace the eedle must travel dowwards to reach the bottom of the well. OpeLyx plate logi If this box is checked ad Use curret MassLyx autosampler bed layout is checked i the OpeLyx Maager program, the the plate at this positio ca oly be used for plate logi o the OpeLyx Logi program. Pressig the butto will move the eedle to the top left vial positio defied by the X, Y ad Vial bottom from probe home positios. If the eedle is ot above the top left vial the the plate will eed movig or the X ad Y values will eed chagig. Pressig the butto will take the eedle to the bottom right vial positio (The software will calculate this from the plate type ad the X ad Y positios defied). This is used to test that the plate will fit o the autosampler, if it does ot the a error message is displayed. The plate will eed chagig or movig or the X ad Y values will eed chagig. Page 35

40 Waters 2700 Fixed Positios This dialog allows the positios of the Ijector Port, Cleaer Statios ad the Waste statio to be defied. It is accessed by selectig Edit Fixed Positios from the Waters2700 meu. Figure 2.24 Fixed Positio Dialog The X ad Y values are the distace the eedle must travel from the Home positio to the required statio. The Z value is the distace the eedle must travel dowwards to reach the required statio. To test that the values etered are valid press the butto. The eedle will travel to the positio specified. To retur the eedle to the Home positio, press the butto. Whe all values are correct press the OK butto. Page 36

41 Waters 2700 Plate Geerator To display the Plate Geerator dialog, select Plate Geerator from the Waters2700 meu. Figure 2.25 Plate Geerator Plate Name The ame of the plate that is curretly beig edited. Rows The umber of vials i a row ad the distace betwee each ceter. Colums The umber of vials i a colum ad the distace betwee each ceter. Offsets Allows alterate vial rows or colums to be offset. Note: Eterig a positive value will shift eve umbered rows to the right ad egative values will shift eve umbered rows to the left. Vial Referece Allows the user to select the way that the vial rows ad colums are refereced, e.g. whether the rows are alphabetical or umerical. Referecig This has three optios XY which refereces the vials A1, B1 etc. Sequetial Discotiuous which umbers the vials 1, 2, 3 across a row, left to right, ad the starts the ext row from the left agai. Sequetial Cotiuous which umbers the vials 1, 2, 3 across a row, left to right, the cotiues umber the ext row, right to left etc. If the Waters 2700 autosampler is used with OpeLyx the the vial referecig must be set to either sequetial cotiuous or sequetial discotiuous. Page 37

42 Priority Check the Horizotal First box if samples are to be acquired horizotally across the plate. If Referecig = X,Y, Horizotal = Letter, Vertical = Number ad Horizotal Priority is checked, this will result i samples beig acquired i the order A1, A2, A3. If the Horizotal Priority box is ot checked samples will be acquired i the order 1A, 1B, 1C etc. If Referecig = sequetial cotiuous or discotiuous ad Horizotal Priority is checked, this will result i samples beig acquired from row 1 the row 2. If the Horizotal Priority box is ot checked samples will be acquired from colum 1 the colum 2 etc. Plate Size The size of the plate to its outside edges. Top Left Vial Offset The measuremet to the ceter of the first vial from the top left corer of the plate. Vial The depth ad diameter values are used for display oly. They appear i the descriptio for a sigle shot logi o the OpeLyx Logi scree. Creatig ad Deletig Waters 2700 Plates To create a ew plate, press the butto. A ew default plate is displayed, chage the Plate Name, eter the appropriate values ad press the save butto or select Save Plate from the Plate meu. New plates are saved to the MassLyx Plates directory. To copy a plate, page through the list of saved plates usig the ad toolbar buttos. The Previous Plate ad Next Plate optios o the Plate meu perform the same operatio. Whe the required plate is displayed chage the Plate Name, eter the appropriate values ad press the save butto or select Save Plate from the Plate meu. New plates are saved to the MassLyx Plates directory. To delete a plate select the plate, by typig the ame i the Plate Name box or by pagig through as above, ad press the delete from the Plate meu. butto or choose Delete Plate Note: All of the spacigs ad the vial sectio are stored i 0.1mm uits. Note: Whe defiig a custom plate for use with a multi-ijector the plate is required to be compatible with the positio of the 8 eedles of the autosampler. The Plate must have eight colums. The positio of the vials should allow all eight eedles to eter a separate vial. There should be o odd or eve offsets for ay of the vial positios. Note: If the Plate curretly selected o the Sample Cofiguratio page is chaged here, the Reset Ijector should be selected from the LC meu to reset commuicatios. Page 38

43 Waters 2700 Meu Prime Syrige This optio is used to remove air from the syrige ad ay tubig coected to it. It repeatedly draws the mobile phase ito the eedle ad flushes it out util the toolbar butto is pressed, or Stop Method is selected from the LC meu, o the Ilet Editor. Note: Before Prime Syrige is selected the toolbar butto appears as ad the meu as Ru Method. Chage Syrige Selectig this optio moves the eedle to a positio where it ca be removed ad replaced. Whe the syrige has bee chaged, Prime Syrige should the be selected to get the eedle ito a state ready for ijectio. Prime Mii-Wash This optio moves the eedle to the waste positio ad pumps the mobile phase through it for the Wash Time defied o the Wash Parameters page. Page 39

44 Waters 2790 Autosampler Note: To cotrol the Waters 2790 autosampler ad pump from the keypad rather tha the MassLyx software the Ilet must be cofigured as Noe. See Cofigurig the Ilet System i the Acquisitio Cotrol Pael chapter. These pages are used to set parameters specific to the Sampler, to access them select Waters2790 AutoSampler from the View meu or press the toolbar butto. Waters 2790 Ijectio Parameters Page Figure 2.26 Ijectio Parameters page Iject Type Select Sequetial or Parallel from the drop dow list box. Sequetial Sample aspiratio occurs at the start of each ijectio cycle, after completio of the previous ijectio. Parallel Sample aspiratio ad loop fill occur cocurretly with other separatio method fuctios for higher throughput. Fill Mode Select Full Loop or Partial Loop from the drop dow list box. Full Loop The autosampler draws i the loop volume the overfill factor umber of times, to esure that the loop is full. Partial Loop The autosampler will draw i the volume specified i the sample list ad cetre it i the loop. Aspirate Air Pre-sample Eter the volume of air to be draw ito the eedle before the sample, to separate it from the previous sample. Rage: 0 to half the loop size. Aspirate Air Post-sample Eter the volume of air to be draw ito the eedle after the sample, to separate it from the ext sample. Rage: 0 to half the loop size. Page 40

45 Loop Off-lie Time For Parallel Ijectio mode, eter the time i miutes whe the ijector valve is switched back from the iject positio to the load positio for the ext sample to be preloaded ito the sample loop. Rage 0.00 to the Ru Time defied o the Pump Mobile Phase page, i miutes. Overfill Factor For full loop mode eter the umber of times to draw the loop volume ito the loop to esure that it is full. Rage 1.0 to 20. Waters 2790 Autosampler Parameters Page Figure 2.27 Autosampler Parameters page Sample Temperature Set If the sample heater is istalled, eter the temperature to heat or cool the sample to. Rage: 4.0 to 40.0 C. Sample Temperature Limit This is the maximum deviatio i sample temperature allowed. If this is exceeded the curret acquisitio will stop ad the LC Status error light, o the MassLyx scree, will tur red. Rage: ±1.0 to ±20.0 C. Loop Size (µl) This is a display oly field showig the volume of the sample loop istalled. Syrige Size This is a display oly field showig the size of the syrige istalled. Draw Depth Adjusts the depth of the eedle tip to accommodate for sedimeted samples or o-stadard vials. A value of 0 correspods to the bottom of the vial. Rage: 0.0 to 20.0 mm. Page 41

46 Draw Speed This determies the rate i microlitres per secod at which sample is extracted ito the autosampler eedle. This should be set accordig to the viscosity of the sample. Select oe of Fast, Normal or Slow from the dropdow list box. The table below shows the draw rate for each selectio usig a 250 µl syrige. Selectio Fast Normal Slow Custom Draw Rate for a 250 µl Syrige 5.0 µl/sec 2.5 µl/sec 1.0 µl/sec Value etered i the Custom Speed box. Seek Well Bottom If this box is checked the, for the first well o a plate, the eedle will automatically seek the bottom of the well before drawig the sample. The depth of the well will be saved by the software ad used as the depth for all other wells o the plate. This will be repeated for the first well o each plate. Note: If a value has bee etered i the Draw Depth field the this operatio will ot be performed. Check Plate Height If this box is checked, for the first ijectio from a plate, a eedle positioig sesor determies the plate height the checks it agaist the Plate Size, Z value defied i the Plate Geerator. Waters 2790 Wash Parameters Page Figure 2.28 Wash parameters page Wash Frequecy Select the wash frequecy from the drop dow list box. Noe Do ot perform a wash. Iject Perform a wash after each ijectio. Well Perform a wash after all samples have bee take from the curret well. Page 42

47 Iject Port Eter the time i secods to wash the iterior of the eedle for. Needle Exterior Eter the time i secods to wash the exterior of the eedle for. Rage: 0 to 99 secods. Wash Cycles Eter the umber of times the Iject Port ad Needle Exterior washes are to be performed. Rage: 0 to 10. Replacemet Volume Eter the volume of wash solvet to leave i the eedle after the wash/flush operatio has bee performed. This volume is the draw through the waste valve ad dispesed ito the sample lie through the eedle. 5DQJHÃÃWRÃÃ O Page 43

48 Waters 2790 Pump Note: To cotrol the Waters 2790 autosampler ad pump from the keypad rather tha the MassLyx software the Ilet must be cofigured as Noe. See Cofigurig the Ilet System i the Acquisitio Cotrol Pael chapter. The Waters Pump pages ca be accessed by selectig Waters2790 Pump from the View meu o the Ilet Editor or by pressig the toolbar butto. Waters 2790 Mobile Phase Page Figure 2.29 Mobile Phase page Solvets Up to four solvets will be displayed depedig upo the system cofiguratio. The total value of all the solvet percetages added together must equal 100%. Solvet Names etered here will be displayed o the Gradiet page. Pump A This displays the remaider percetage after the solvet percetages have bee set for the other eabled pumps. Pump B, C ad D These ca either be eabled or disabled by checkig the box ext to the idividual pumps. The values ca be set to the required percetage of flow delivery. Flow Eter the total iitial flow rate of the system. Rage: to ml/mi. Ramp Eter the time (i miutes) for the solvet delivery system to reach the maximum system flow rate (10 ml/mi). This limits the rate of chage of the flow rate to protect the colum from potetially damagig sudde chages i pressure. Rage: 0.01 to 30 miutes. Recommeded miimum settig: 0.5 mi. Eter Low Pressure Limit ad High Pressure Limit values as required. If the pressure falls outside these limits the curret acquisitio will stop ad the LC Status error light, o the MassLyx scree, will tur red. Low Pressure Rage: 0 to 310 bar. High Pressure Rage: 0 to 345 bar. Page 44

49 Degasser Select oe of Off, Normal or Cotiuous from the drop dow list box. Off The degasser is always off. Normal The degasser cycles o ad off. Cotiuous The degasser is always o. Stroke Legth This sets the volume of solvet delivered for each pisto stroke. Select the required optio from the drop dow list box. If Auto is selected the the volume is automatically adjusted to provide optimal performace for the selected solvet flow rate, otherwise the volume selected will be used. Ru Time Eter the time i miutes that the method will ru from the poit of ijectio. Note: Ru time is for the solvet delivery system oly. Detectors have idepedet ru times. The MS method (Sca Fuctio Editor) ru time must be greater tha all other ru times. Waters 2790 Colum Page Figure 2.30 Colum page Positio This field allows the colum to be selected for the method. The optios available will deped o the colum setup o the Waters 2790 Separatios Module. If oly oe colum is istalled the this box will display Colum 1 ad caot be chaged. For other cofiguratios this box will allow the selectio of a colum (betwee 1 ad 6 depedig o cofiguratio) or No chage from the drop dow list box. Selectig a umbered colum will use this colum for the method, selectig No Chage will use the colum defied i the last method used to acquire a sample. See the Waters 2790 Separatios Module Operator s Guide for more iformatio o the colum selectio valve. Page 45

50 Equilibrium Time Eter the time required to reach equilibrium (i.e. ru i iitial coditios), before performig a ijectio, after a colum chage. Rage: 0.00 to miutes. Temperature Set Eter the target operatig temperature for the optioal colum heater. This value must be at least 5 C above ambiet. Rage: 20 to 60 C. Temperature Limit Eter the maximum deviatio i colum temperature allowed. If this is exceeded the curret acquisitio will stop ad the LC Status error light, o the MassLyx scree, will tur red. Rage: ±1 to ±20 C. Waters 2790 Rapid Equilibratio Page Figure 2.31 Rapid Equilibratio page Path Select the path to be used for flushig solvet durig rapid equilibratio from the drop dow list box. Waste, Off or Colum 1 to Colum 6 depedig o the istrumet cofiguratio. Flow Eter the system equilibratio flow rate. Rage: 0.00 to ml/mi. Time Eter the legth of time (i miutes) to equilibrate. Rage 0.00 to miutes. Re-equilibratio Time Eter the time that colum should be maitaied at iitial flow/compositio coditios after completio of a gradiet ru. This delay is imposed o a per ijectio basis if defied. Pre-colum Volume Eter the volume of solvet to pump through the colum betwee the time the gradiet starts ad the time of ijectio. Rage: 0.0 to µl. Page 46

51 Waters 2790 I/O Page Figure 2.32 I/O page Switch Iitial Coditios Select the state that switches 1 to 4 should be i iitially, from the drop dow list box. At the begiig of each ijectio cycle each switch returs to the state defied here. Available choices: O Turs o a cotact closure that triggers a exteral or iteral evet. With this fuctio, the cotact closure remais closed util a Off fuctio is set. Off Turs off the cotact closure for the evet. With this fuctio, the cotact closure is broke. Pulse Trasmits a sigle O/Off pulse. The cotact closure is maitaied for the umber of miutes set i the Pulse Width field o the Evets page. Rage: 0.01 to sec. Toggle Chages the curret state of the switch. No Chage Leaves the switch i its curret state. Chart Output Settig Select Flow Rate, System Pressure, %A, %B, %C, %D, Colum Temperature or Sample Temperature from the drop dow list box. The Aalog output sigals are set through the termials o the back of the 2790, to a optioal aalog device such as a strip chart recorder. If, for example, System Pressure is selected the recorder will chart the system pressure while the method is beig ru. Page 47

52 Waters 2790 Gradiet Page Figure 2.33 Gradiet page This page allows a gradiet to be etered ad edited. If you wish to operate i isocratic mode the eter parameters o the Mobile Phase page ad esure that the timetable is empty. Time (mis) Specifies the time at which the specified coditios (%A to %D, Flow, ad Curve) for the row should take effect. Make sure the Time for the first row is set to 0.00, to establish iitial coditios for the gradiet ru. The rage for rows other tha row 1is 0.01 to miutes. Solvet A % - Solvet D% Specifies the percetage of solvet flow from each reservoir. For each row the total of all solvets must equal 100%. Rage: 0 to 100%. Flow (ml/mi) Specifies the total flow rate for the solvet delivery system. Rage: 1 to 10 ml/mi. Note: If colum equilibratio, rapid equilibratio or wet prime are performed the the flow rate will retur to the value defied o the Mobile Phase page. If they are ot performed the the flow rate will stay at the value defied for the last etry i the Gradiet Table. To retur to the iitial flow rate a etry must be added to the ed of the table settig the value to that defied o the Mobile Phase page. Curve This sets the rate at which the solvet is to chage to the ew proportios ad/or flow rates. Curves are specified by umber. Available choices: 1 to 11. Page 48

53 Curve Number Effect 1 Immediately goes to specified coditios 2 to 5 Covex 6 Liear 7 to 10 Cocave 11 Maitais start coditio util ext step Curve Profiles Waters 2790 Gradiet Table Operatio To add a gradiet, eter values i the relevat boxes ad press the toolbar butto. Up to 15 rows ca be added to the table. Note: The first etry must have a time of 0. To delete a sigle gradiet click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To modify a gradiet, select the required etry i the timetable. The values will the be displayed i the edit boxes ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig will result i a ew etry beig created i the timetable. Page 49

54 Waters 2790 Evets Page Figure 2.34 Evets page Use the Evet Table to program up to 16 evets (both exteral ad iteral). The exteral evets are triggered by four cotact closures (relays) through output termials (S1 S4) o the 2790 Separatios Module. The iteral evets are used to cotrol the sample compartmet temperature, colum heater temperature ad colum chage. Evets ca be triggered more tha oce ad multiple evets ca be triggered simultaeously. Time Eter the time at which the evet starts. Evet rows are sorted automatically by time. Note: Differet evets ca be programmed to occur at the same time. Rage 0.00 to the Ru Time defied o the Mobile Phase page, i miutes. Evet Eter the type of evet sigal: oe of the four TTL-level output switches (S1-S4), or oe of the iteral evets (colum heater temperature, sample compartmet or colum chage). Switch 1 to Switch 4 Correspods to termial strip positios S1 to S4 o the rear of the 2790 module. Activatig a Switch evet triggers a cotact closure for cotrollig a exteral device. After selectig a switch evet, set a state for the switch by selectig O, Off, Toggle, Pulse Width or No Chage. This state appears i the Actio colum of the table (see Switch States, below). Note: If Pulse is selected the duratio of the pulse must be etered i the Width (mi) field. Set Temperature (Colum or Sample) Specifies the temperature of a optioal colum heater, or a optioal sample compartmet heater/cooler. After selectig this evet, select Colum or Sample ad eter the required Temperature i C. Note: Whe a Colum Temperature evet occurs, the temperature of the colum heater chages from the value set i the Colum page to the value set for the evet. Whe the evet times out, the temperature chages back to the Colum page value. Colum rage: 20 to 60 C. Sample rage: 4 to 40 C. Colum Chage Specifies a colum chage operatio for the 2790 module, as describe o the Colum page. Page 50

55 Switch States O Turs o a cotact closure that triggers a exteral or iteral evet. With this fuctio, the cotact closure remais closed util a Off fuctio is set. Off Turs off the cotact closure for the evet. With this fuctio, the cotact closure is broke. Toggle Chages the curret state of the switch. Pulse Trasmits a sigle O/Off pulse. The cotact closure is maitaied for the umber of secods that defied i the Value colum. Rage: 0.01 to sec. No Chage Leaves the switch i its curret state. Waters 2790 Evet Table Operatio To add a evet, type i a time, select a evet from the drop dow list box, select a actio ad press the toolbar butto. Up to 16 evets ca be programmed. To delete a sigle evet click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To modify a evet select the required etry i the timetable. The values will the be displayed i the edit boxes ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig will result i a ew etry beig created i the timetable. Page 51

56 Waters 2790 Method Type Page Figure 2.35 Method Type page This page is used for creatig ormal, pre ad post ru methods. First create a Normal method ad save the file, the create ay pre or post ru methods (savig them uder differet ames). These methods ca the be defied i the Ilet Preru ad Ilet Postru colums of the Sample List. Method Type Select the type of method to create. The Parameters sectio will be updated to show the parameters required for the selected method. Normal Creates a ormal method. No extra parameters eed to be defied. For all other method types see the Waters 2790 Meu, o page 53 for details of the parameters required. Page 52

57 Waters 2790 Meu Waters 2790 Wet Prime Figure 2.36 Wet Prime dialog Wet Prime A wet prime should be performed whe chagig the solvet i the system to flush out the previous solvet. The ew solvet is pumped through the tubig ad the Prime port of the Iject valve to waste. Eter the Flow rate, Time ad the Percetage of solvets to use the press the Prime butto. Waters recommed that the wet prime is started usig the solvet with the lowest viscosity to help purge air from the lies, especially if the i-lie vacuum degasser is istalled. Note: If the solvet lies are dry the a dry prime must be performed before a wet prime. See the Waters 2790 Separatio Module Operator s Guide for more iformatio o performig a dry prime. Waters 2790 Equilibrate Figure 2.37 Equilibrate dialog Equilibrate Equilibrates the system usig the parameters defied o the Mobile Phase page. Eter the Time to equilibrate the system for ad press OK. The time eeded to equilibrate the system will deped o evirometal ad applicatio-specific factors. Page 53

58 Waters 2790 Coditio Colum Figure 2.38 Coditio Colum dialog Coditio Colum Rus solvet through the colum without ijectig samples or ruig the Evets table. Solvet is delivered to the Colum defied o the Colum page, usig the Gradiet Table defied o the Gradiet page. Eter the Time i miutes to coditio the colum for ad press OK. Esure that the time is equal to or greater tha the Time of the last etry i the Gradiet Table (defied o the Gradiet page) plus the Re-equilibratio Time (defied o the Rapid Equilibratio page). Waters 2790 Wash Needle Figure 2.39 Wash Needle dialog Wash Needle Washes the iject port, ad both the iterior ad exterior of the eedle with wash solvet, ad the fills the eedle with fresh solvet. Eter the umber of wash Cycles to perform ad the Volume of wash solvet to use the press OK. Waters recommed a volume of 600 µl. Waters 2790 Refresh Syrige Figure 2.40 Refresh Syrige dialog Refresh Syrige Refills the syrige with fresh, degassed, purge solvet. Eter the umber of Cycles ad the replacemet Volume ad press OK. Waters recommed 12 Cycles ad a volume of 600 µl. Page 54

59 Waters 2790 Plate Geerator To display the Plate Geerator dialog, select Plate Geerator from the Waters2790 meu. Figure 2.41 Plate Geerator Plate Name The ame of the plate that is curretly beig edited. Rows The umber of vials i a row ad the distace betwee each ceter. Colums The umber of vials i a colum ad the distace betwee each ceter. Offsets Allows alterate vial rows or colums to be offset. Note: Eterig a positive value will shift eve umbered rows to the right ad egative values will shift eve umbered rows to the left. Vial Referece Allows the user to select the way that the vial rows ad colums are refereced, e.g. whether the rows are alphabetical or umerical. Horizotal Sets the horizotal axis of the plate as either alphabetic (ABC) or umeric (123), whe usig XY referecig. Vertical Sets the vertical axis of the plate as either alphabetic (ABC) or umeric (123), whe usig XY referecig. Page 55

60 Referecig This has three optios XY which refereces the vials A1, B1 etc. Sequetial Discotiuous which umbers the vials 1, 2, 3 across a row, left to right, ad the starts the ext row from the left agai. Sequetial Cotiuous which umbers the vials 1, 2, 3 across a row, left to right, the cotiues umber the ext row, right to left etc. If the Waters 2790 autosampler is used with OpeLyx the the vial referecig must be set to either sequetial cotiuous or sequetial discotiuous. Priority Check the Horizotal First box if samples are to be acquired horizotally across the plate. If Referecig = X,Y, Horizotal = Letter, Vertical = Number ad Horizotal Priority is checked, this will result i samples beig acquired i the order A1, A2, A3. If the Horizotal Priority box is ot checked samples will be acquired i the order 1A, 1B, 1C etc. If Referecig = sequetial cotiuous or discotiuous ad Horizotal Priority is checked, this will result i samples beig acquired from row 1 the row 2. If the Horizotal Priority box is ot checked samples will be acquired from colum 1 the colum 2 etc. Plate Size The size of the plate to its outside edges. Top Left Vial Offset The measuremet to the ceter of the first vial from the top left corer of the plate. Vial The depth ad diameter values are used for display oly. They appear i the descriptio for a sigle shot logi o the OpeLyx Logi scree. Creatig ad Deletig Waters 2790 Plates To create a ew plate, press the butto. A ew default plate is displayed, chage the Plate Name, eter the appropriate values ad press the save butto or select Save Plate from the Plate meu. New plates are saved to the MassLyx Plates directory. To copy a plate, page through the list of saved plates usig the ad toolbar buttos. The Previous Plate ad Next Plate optios o the Plate meu perform the same operatio. Whe the required plate is displayed chage the Plate Name, eter the appropriate values ad press the save butto or select Save Plate from the Plate meu. New plates are saved to the MassLyx Plates directory. To delete a plate select the plate, by typig the ame i the Plate Name box or by pagig through as above, ad press the delete from the Plate meu. butto or choose Delete Plate Note: All of the spacigs ad the vial sectio are stored i 0.1 mm uits. Note: Whe defiig a custom plate for use with a multi-ijector the plate is required to be compatible with the positio of the 8 eedles of the autosampler. Page 56

61 The Plate must have eight colums. The positio of the vials should allow all eight eedles to eter a separate vial. There should be o odd or eve offsets for ay of the vial positios. Note: If the Plate curretly selected o the Sample Cofiguratio page is chaged here, the Reset Ijector should be selected from the LC meu to reset commuicatios. Note: All of the spacigs ad the vial sectio are stored i 0.1 mm uits. Vial Referecig Examples The followig tables show four examples of vial referecig for a simplified 4 3 vial plate A 1,A 2,A 3,A 4,A B 1,B 2,B 3,B 4,B C 1,C 2,C 3,C 4,C Horizotal: 123 Vertical: ABC Referecig: XY Priority: Horizotal First Checked A A,1 A,2 A,3 A,4 B B,1 B,2 B,3 B,4 C C,1 C,2 C,3 C,4 Horizotal: 123 Vertical: ABC Referecig: XY Priority: Horizotal First NOT Checked A B C Horizotal: N/A Vertical: N/A Referecig: Sequetial Discotiuous Priority: Horizotal First Checked A B C Horizotal: N/A Vertical: N/A Referecig: Sequetial Cotiuous Priority: Horizotal First NOT Checked Page 57

62 Waters 2790 Bed Layout Use the Bed Layout Editor to defie the type, umber, ad locatio of the well plates o the 2790 plate carrier. To access the Bed Layout Editor, select Bed Layout from the Waters2790 meu. Figure 2.42 Bed Layout dialog Bed Layouts Lists the available Bed Layouts. Curret Layout Specifies the bed layout curretly i use. To Delete A Bed Layout (Waters 2790) Highlight the bed layout to delete ad press the butto. A dialog box will ask you to cofirm the deletio. Press the OK butto to delete the bed layout. Note: You caot delete the bed layout which is selected as the Curret Layout. To Create A New Bed Layout (Waters 2790) 1. Highlight a bed layout similar to the oe you wat to create ad press the butto to create a ew layout. The layout appears i the Bed Layouts list as the same ame with a 1 at the ed, for example Six Positio Microtiter1. 2. To chage the ame of the layout, type the ew ame ito the Bed Layouts text box ad press the list box. butto. The ame is updated i the Bed Layouts 3. If the plate positio or type eeds chagig select the Modify Bed Layout tab. Note: New bed layouts are saved to the MassLyx Racks directory. Page 58

63 To Modify a Bed Layout (Waters 2790) Use the Modify Bed Layout page to modify a existig bed layout. To access the Modify Bed Layout page, click the Modify Bed Layout tab. The Modify Bed Layout page shows a graphical represetatio of the selected bed layout. There are four plate positios i the Figure 2.43 Modify Bed Layout dialog Click the plate that you wat to chage to display the Plate Positio ad Type dialog. Figure 2.44 Plate Positio ad Type dialog This dialog allows you to select a ew plate from a list of possible optios, ad chage its actual positio o the bed. Select the plate type you wat to use i the bed layout, the click OK. OpeLyx plate logi If this box is checked ad Use curret MassLyx autosampler bed layout is checked i the OpeLyx Maager program, the the plate at this positio ca oly be used for plate logi o the OpeLyx Logi program. Page 59

64 Waters CapLC System Status Pages The System Status pages display iformatio about the state of the machie beig cotrolled. These pages ca be accessed withi the Ilet Editor by selectig Status from the View meu or by pressig the toolbar butto. Waters CapLC Solvet Status Page Figure 2.45 Solvet Status page Idicators The Ruig, Pump O ad Ijector Cycle idicators at the left of the scree give iformatio o the curret status of the LC system. The OK ad Ready Idicators become illumiated i red if the LC System has a error. Click o the red idicators to display more iformatio o the cause of the malfuctio. Time This displays how log the method has bee ruig. Flow This displays the curret flow rate as retured by the istrumet. Pressure This displays the curret pressure i the istrumet. To the right of the Time, Flow ad Pressure fields is a display of the solvet percetages at which the LC System is curretly operatig. Colum This displays the curret temperature of the colum. Sample Temp This displays the curret temperature of the sample. Page 60

65 Waters CapLC Valve Status Page Figure 2.46 Valve Status page Figure 2.47 Valve Status page This page shows what positio the Valves are curretly set to, if istalled. See the Waters Cap LC Users Guide for details of the valves. Page 61

66 Waters CapLC PDA Status Page Figure 2.48 PDA Status page Curret State This displays the curret state of the PDA Detector. Sca Number Whe acquirig diode array data this displays the umber of scas curretly acquired. Page 62

67 Waters CapLC Pump The Waters Pump pages ca be accessed by selectig WatersCapLC Pump from the View meu o the Ilet Editor or by pressig the toolbar butto. Waters CapLC Iitial Coditios Page Figure 2.49 Iitial Coditios page Solvet Select A1 to A3 Select the solvet to deliver through Pump A. Solvet B ad C Eter the percetage of solvet flow from pump B ad/or C (if istalled). Solvet Name Eter the ame of the solvet i the correspodig solvet reservoir. Flow Eter the total flow rate for the system i µl/mi. Ru Time Eter the legth of time (i miutes) util the ext ijectio occurs. Note: Ru time is for the pump ad autosampler oly. Detectors have idepedet ru times. The MS method ru time must be greater tha all other ru time (see The Fuctio List Editor chapter for details). Note: If you are ruig a gradiet or settig timed evets, make sure you set the iitial coditios Ru Time to a value greater tha or equal to the greatest Time value i the Gradiet or Timed Evets Table. Low Pressure Eter the low-pressure limit for the system. If the system pressure falls below this limit, the flow stops ad the LC Status error light turs red. Rage: 0 to 4500 psi. High Pressure Eter the high-pressure limit for the system. If the system pressure exceeds this limit, the flow stops ad the LC Status error light turs red. Rage: 0 to 5000 psi. Page 63

68 Temperature Eter the target operatig temperature for the optioal colum heater. This value must be at least 5 C above ambiet. Rage: 20 to 60 C. Miimum settig: 5 C above ambiet. Rage Eter the maximum allowable temperature deviatio from the value set for the colum heater temperature. If the colum heater temperature deviates beyod the specified rage, the ru stops ad the LC Status error light turs red. Rage: ±0.0 to ±10.0 C. Name Eter the ame of the istalled colum. Waters CapLC Gradiet Page Figure 2.50 Gradiet page Use the Gradiet Table to defie coditios for a gradiet ru. For each row i the Gradiet Table, defie the percet compositio of up to four solvets that are to be delivered at the desired flow rate for the specified time. Eter the umber of the gradiet curve required. This defies how chages to solvet percetages ad flow rates take place over the elapsed time of each gradiet segmet (the time that elapses betwee the start time of oe row ad the start time of the ext row). Note: For a isocratic ru, set the solvet percetages, ru time ad flow o the Iitial Coditios page. Do ot add ay rows to the Gradiet Table. Waters CapLC Gradiet Table Parameters Time (mis) Specifies whe the coditios (%A-%D, Flow, ad Curve) for the row take effect. Make sure the Time for the first row is set to 0.00 to establish iitial coditios for the gradiet ru. Rage for rows other tha row 1: 0.01 to miutes. B% ad C% Specifies the percetage of solvet flow from each reservoir. For each row, the total of all solvets must equal 100%. Rage: 0 to 100%. Note: Percet flow for reservoir A is ot displayed. Percet A is calculated as: 100% - (B% + C%) Page 64

69 Flow (µl/mi) Specifies the total flow rate for the system. Curve Specifies the rate of chage of solvet compositio ad flow rate over time, based o the curve umber ad the legth of the gradiet segmet. For more iformatio, see Gradiet Curves, below. Waters CapLC Gradiet Table Operatio To add a gradiet, eter a time, percetage, flow rate ad curve umber i the relevat boxes ad press the time of 0. toolbar butto. Note: The first etry must have a To delete a sigle gradiet click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To modify a gradiet select the required etry i the timetable. The values will the be displayed i the edit boxes above the timetable, ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig Gradiet Curves will result i a ew etry beig created i the timetable. Curve Number Effect 1 Immediately goes to specified coditios 2 to 5 Covex 6 Liear 7 to 10 Cocave 11 Maitais start coditio util ext step Curve Profiles Page 65

70 Waters CapLC Iitial Evets Page Figure 2.51 Iitial Evets page Use the Iitial Evets page to set the iitial coditio of the two cotact-closure output switches, the iitial positio of the stream select valve, ad the iitial positio of the optioal diverter valve. Note: To chage the settigs of these switches ad valves durig a ru, use the Timed Evets Table. Switch 1 ad 2 Select the iitial state of cotact-closure Switch 1 ad 2 from the drop dow list boxes. Stream Select Select the iitial state of the stream select valve (Positio 1 or 2) from the drop dow list box. Diverter Select the iitial positio of the optioal diverter valve (System or Vet) from the drop dow list box. Page 66

71 Waters CapLC Timed Evets Page Figure 2.52 Timed Evets page Use the Evet Table to program up to 16 evets (both exteral ad iteral). The exteral evets are triggered by four cotact closures (relays) through output termials (S1 S4) o the 2790 Separatios Module. The iteral evets are used to cotrol the sample compartmet temperature, colum heater temperature, ad to prime ad flush the 2790 Separatios Module. Evets ca be triggered more tha oce ad multiple evets ca be triggered simultaeously. Waters CapLC Evet Table Parameters Time Eter the time (after ijectio) at which the evet starts. Evet rows are sorted automatically by time. Note: Differet evets ca be programmed to occur at the same time. Rage: 0.00 to mi. Evet Select the type of evet sigal: oe of the two cotact-closure output switches (Switch1or Switch 2), or oe of the iteral evets (Set temperature, Stream Select or Vet Valve). Choose from these evet types to program up to 16 evets. Note: The same evet ca be programmed more tha oce. Available choices: Switch 1 ad 2 Correspods to termial strip positios S1 ad S2 o the rear of the uit. Activatig a Switch evet triggers a cotact closure for cotrollig a exteral device. Select a switch evet ad a state for the switch (O, Off, Toggle, Pulse or No Chage). This state appears i the Actio colum of the table (see Switch States, below). Note: If Pulse is selected for a switch state the duratio of the pulse must be etered i the Width (mi) field. Page 67

72 Set Temperature Specifies the temperature of a optioal colum heater. If Set Temperature is selected for a switch state the temperature i ( C) must be etered i the Colum Temperature field. Note: Whe this evet occurs, the temperature of the colum heater chages from the value set o the Iitial Coditios page to the value set for the evet. Whe the evet times out, the temperature returs to the value o the Iitial Coditios page value. Stream Select (1 or 2) Specifies the positio of the stream select valve. Vet Valve (System or Vet) Specifies the positio of the vet valve. Waters CapLC Switch States O Turs o a cotact closure that triggers a exteral or iteral evet. With this fuctio, the cotact closure remais closed util a Off fuctio is set. Off Turs off the cotact closure for the evet. With this fuctio, the cotact closure is broke. Toggle Chages the curret state of the switch. Pulse Trasmits a sigle O/Off pulse. The cotact closure is maitaied for the time etered i the Width box. Rage: 0.01 to miutes. No Chage Leaves the switch i its curret state. Waters CapLC Evet Table Operatio To add a evet, eter a time, evet, actio ad value i the relevat boxes ad press the toolbar butto. Note the first etry must have a time of 0. To delete a sigle evet click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To modify a evet select the required etry i the timetable. The values will the be displayed i the edit boxes, ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig will result i a ew etry beig created i the timetable. Page 68

73 Waters CapLC Chart Recordig Page Figure 2.53 Chart Recordig page Use the Chart Recordig page to select up to two aalog sigals to be output to a exteral device such as a itegrator or strip-chart recorder. Select oe of the followig the sigals to output, from the drop dow list box. Noe Flow Pressure Percet A, B, or C Colum temperature Note: To record a sigal, you eed to coect each exteral device to the appropriate Chart Out termial pair o the rear of the uit. Refer to the Waters CapLC System Istallatio ad Maiteace Guide for istallatio ad specificatio details. Page 69

74 Waters CapLC Method Type Page Figure 2.54 Method Type page Method Type Specifies the type of method to create. Select oe of: Normal Method The Method Type used for stadard ijectios. Esure that the Method Type is set to Normal uless you are performig oe of the procedures listed below. Colum Coditio Method Rus solvet through the colum without ijectig samples or ruig the Evets table. Solvet is delivered usig the gradiet table specified i the Gradiet page. Equilibrate Method Delivers solvets ad maitais solvet parameters usig the values defied o the Iitial Coditios page. Wet Prime Replaces solvet i the tubig with fresh solvet from the reservoirs through the Prime port of the iject valve to waste. Use a Wet Prime Method whe chagig the solvets i the system. Check the boxes for the solvet lies to prime, ad the umber of loop volumes to use. Waters recommeds startig the wet prime usig the solvet with the lowest viscosity to help purge air from the lies, especially if the i-lie vacuum degasser is istalled. Note: If the solvet lies i the CapLC are dry, you must perform the dry prime procedure before performig a wet prime. Page 70

75 Waters CapLC Autosampler These pages are used to set parameters specific to the Autosampler, to access them select WatersCapLC AutoSampler from the View meu or press the butto. toolbar Waters CapLC AutoSampler Page Figure 2.55 CapLC AutoSampler page Draw Height Adjusts the depth of the eedle tip to accommodate for sedimeted samples. A value of 0 correspods to the top of the plate carrier. Rage: 0 to 40 mm. Draw Speed Select the draw rate of the syrige from the drop dow list box. The differet rates accommodate for samples of varyig viscosity. The rate for each selectio is depedet o the size of the istalled syrige. The table below shows the draw rates for each selectio. Draw Rate for 25 µl Syrige Draw Rate for 100 µl Syrige Draw Rate for 250 µl Syrige Draw Rate for 500 µl Syrige Fast 94 µl/mi 375 µl/mi 940 µl/mi 1875 µl/mi Normal 63 µl/mi 250 µl/mi 625 µl/mi 1250 µl/mi Slow 32 µl/mi 375 µl/mi 315 µl/mi 625 µl/mi Page 71

76 Ijectio Type Select oe of the followig ijectio types from the drop dow list box: Full Loop The sample loop is completely filled. Partial Loop The sample loop is partially filled with the volume defied i the Sample List. The value i the Sample List must ot exceed the Full Loop volume. µl Pickup The sample loop is filled with oly the amout of sample to be ijected (resultig i o sample loss). Sample is trasported ito the loop by trasport liquid (mobile phase) from the trasport vial. Maual Specifies that the maual ijector is used (the autosampler is disabled). Switchig the maual ijector to the Iject positio iitiates ay programmed gradiets ad/or timed evets. Flush Volume Eter the volume (i microliters) of sample take from a vial before the loop is filled with sample. This flushes out previous samples. Head Space Pressure If this box is checked the prepucturig eedle will put approximately 0.5 bar of pressure o the sample to stop formatio of air or vapour bubbles. Eable this parameter oly whe usig sample vials with air-tight caps. Air Space Segmet If this box is checked a air segmet is added to the frot of the flush volume to miimize dilutio ad badspreadig ad reduce the amout of flush volume required. I Full ad Partial Loop modes, the air segmet is flushed to waste; i µl Pickup mode, the air segmet is ijected. Disable this parameter if the air segmet causes problems i µl Pickup mode. High Efficiecy Mode If this box is checked the sample loop will be take out of the flow stream after the sample has bee flushed, but before the gradiet frot reaches the ijectio valve. Wash Volume Eter the volume (i microliters) of wash solvet used to clea the eedle ad buffer tubig. Sample Temperature Eter the target operatig temperature for the optioal sample heater/cooler. Rage: 4.0 to 40.0 C. Temperature Limit Eter the maximum allowable temperature deviatio from the value set for the Sample Temperature. If the sample temperature deviates beyod the specified rage, the LC Status error light turs red ad the ru stops. Rage: ±1.0 to ±20.0 C. Page 72

77 Waters CapLC Mix Method Page Figure 2.56 Mix Method page Mix Delay Eter the delay time before mixig, i miutes. A value of 0 correspods to the top of the plate carrier. Rage: 0 to 99.9 mis. Mix Cycles Eter the umber of times to perform the Mix operatio. Reaget Positio 1 Eter the positio of the first reaget to mix. Volume Reaget 1 Eter the volume of reaget 1 to mix. Reaget Positio 2 Eter the positio of the secod reaget to mix. Volume Reaget 2 Eter the volume of reaget 2 to mix. Page 73

78 Waters CapLC Bed Layout Use the Bed Layout Editor to defie the type, umber, ad locatio of the well plates o the CapLC plate loader. To access the Bed Layout Editor, select Bed Layout from the Cap LC Tools meu. Figure 2.57 Bed Layout Dialog Bed Layouts Lists the available Bed Layouts. Curret Layout Specifies the bed layout curretly i use. To Delete A Bed Layout (Waters CapLC) Highlight the bed layout to delete ad press the butto. A dialog box will ask you to cofirm the deletio. Press the OK butto to delete the bed layout. Note: You caot delete the bed layout which is selected as the Curret Layout. To Create A New Bed Layout (Waters CapLC) 1. Highlight a bed layout similar to the oe you wat to create ad press the butto. The layout appears i the Bed Layouts list as the same ame with a 1 at the ed, for example Six Positio Microtiter1. 2. To chage the ame of the layout, type the ew ame ito the Bed Layouts text box ad press the list box. butto. The ame is updated i the Bed Layouts 3. If the plate positio or type eeds chagig select the Modify Bed Layout tab. Note: New bed layouts are saved to the MassLyx Racks directory. Page 74

79 To Modify a Bed Layout (Waters CapLC) Use the Modify Bed Layout page to modify a existig bed layout. To access the Modify Bed Layout page, click the Modify Bed Layout tab. The Modify Bed Layout page shows a graphical represetatio of the selected bed layout. Figure 2.58 Modify Bed Layout Dialog Click o the plate that you wat to chage to display the Plate Positio ad Type dialog. Figure 2.59 Plate Positio ad Type Dialog This dialog allows you to select a ew plate from a list of possible optios, ad chage its actual positio o the bed. Select the plate type to use i the bed layout, the click OK. OpeLyx plate logi If this box is checked ad Use curret MassLyx autosampler bed layout is checked i the OpeLyx Maager program, the the plate at this positio ca oly be used for plate logi o the OpeLyx Logi program. Page 75

80 Other Bed Layout Optios (Waters CapLC) 1. To apped a ew colum, press the butto. 2. To delete the curret colum press the butto. 3. To isert a colum, click o the colum before which you wat to isert ad press the butto. Note: The colum iserted will have the same umber of rows as the colum highlighted. 4. Note: The umber of rows i a colum caot be chaged ad so the butto is greyed out. Page 76

81 Waters CapLC Plate Geerator To display the Plate Geerator dialog, select Plate Geerator from the Cap LC Tools meu. Figure 2.60 Plate Geerator Plate Name The ame of the plate that is curretly beig edited. Rows The umber of vials i a row ad the distace betwee each ceter. Colums The umber of vials i a colum ad the distace betwee each ceter. Offsets Allows alterate vial rows or colums to be offset. Note: Eterig a positive value will shift eve umbered rows to the right ad egative values will shift eve umbered rows to the left. Vial Referece Allows the user to select the way that the vial rows ad colums are refereced, e.g. whether the rows are alphabetical or umerical. Horizotal Sets the horizotal axis of the plate as either alphabetic (ABC) or umeric (123), whe usig XY referecig. Default: umeric. Vertical Sets the vertical axis of the plate as either alphabetic (ABC) or umeric (123), whe usig XY referecig. Default: alphabetic. Referecig This has three optios XY which refereces the vials A1, B1 etc. Sequetial Discotiuous which umbers the vials 1, 2, 3 across a row, left to right, ad the starts the ext row from the left agai. Sequetial Cotiuous which umbers the vials 1, 2, 3 across a row, left to right, the cotiues umber the ext row, right to left etc. Page 77

82 If the Waters CapLC autosampler is used with OpeLyx the the vial referecig must be set to either sequetial cotiuous or sequetial discotiuous. Priority Check the Horizotal First box if samples are to be acquired horizotally across the plate. If Referecig = X,Y ad Horizotal First is checked, the the horizotal value be read first whe referecig a vial (1,A). If Horizotal First is ot selected, the the vertical value be read first whe referecig a vial (A,1). Default: Horizotal First selected. If Referecig = Sequetial Cotiuous or Discotiuous ad Horizotal First is checked, the vials will be umbered horizotally. This will result i samples beig acquired from row 1 the row 2. If Horizotal First is ot checked, the vials will be umbered vertically. This will result i samples beig acquired from colum 1 the colum 2 etc. Default: Horizotal First selected. Plate Size The size of the plate to its outside edges. Top Left Vial Offset The measuremet to the ceter of the first vial from the top left corer of the plate. Vial The depth ad diameter values are used for display oly. They appear i the descriptio for a sigle shot logi o the OpeLyx Logi scree. Creatig ad Deletig Waters CapLC Plates To create a ew plate press the butto. A ew default plate is displayed, chage the Plate Name, eter the appropriate values ad press the save butto or select Save Plate from the Plate meu. New plates are saved to the MassLyx Plates directory. To copy a plate, page through the list of saved plates usig the ad toolbar buttos. The Previous Plate ad Next Plate optios o the Plate meu perform the same operatio. Whe the required plate is displayed chage the Plate Name, eter the appropriate values ad press the save butto or select Save Plate from the Plate meu. New plates are saved to the MassLyx Plates directory. To delete a plate select the plate, by typig the ame i the Plate Name box or by pagig through as above, ad press the delete from the Plate meu. butto or choose Delete Plate Note: All of the spacigs ad the vial sectio are stored i 0.1 mm uits. Note: Whe defiig a custom plate for use with a multi-ijector the plate is required to be compatible with the positio of the 8 eedles of the autosampler. The Plate must have eight colums. The positio of the vials should allow all eight eedles to eter a separate vial. There should be o odd or eve offsets for ay of the vial positios. Page 78

83 Note: If the Plate curretly selected o the Sample Cofiguratio page is chaged here, the Reset Ijector should be selected from the LC meu to reset commuicatios. Note: All of the spacigs ad the vial sectio are stored i 0.1 mm uits. Vial Referecig Examples The followig tables show four examples of vial referecig for a simplified 4 3 vial plate A 1,A 2,A 3,A 4,A B 1,B 2,B 3,B 4,B C 1,C 2,C 3,C 4,C Horizotal: 123 Vertical: ABC Referecig: XY Priority: Horizotal First Checked A A,1 A,2 A,3 A,4 B B,1 B,2 B,3 B,4 C C,1 C,2 C,3 C,4 Horizotal: 123 Vertical: ABC Referecig: XY Priority: Horizotal First NOT Checked A B C Horizotal: N/A Vertical: N/A Referecig: Sequetial Discotiuous Priority: Horizotal First Checked A B C Horizotal: N/A Vertical: N/A Referecig: Sequetial Cotiuous Priority: Horizotal First NOT Checked Page 79

84 Waters CapLC Plate Loader To display the Plate Loader dialog, select Plate Loader from the Cap LC Tools meu. Figure 2.61 Plate Loader The Plate Loader dialog is used whe a plate eeds to be chaged. Page 80

85 Waters CapLC PDA Detector This page is used to set parameters specific to the UV detector, to access it select WatersCapLC PDA Detector from the View meu or press the toolbar butto. Figure 2.62 CapLC PDA Detector Cofiguratio page Start Wavelegth Eter the wavelegth at which to start acquirig data. Rage (with Resolutio set to 1.2): m to m. Rage (at all other Resolutio settigs): (Resolutio/2) to (Resolutio/2). Default: 200 m. Ed Wavelegth Eter the wavelegth at which to stop acquirig data. Rage with Resolutio set to 1.2: Start Wavelegth to m. Rage at all other Resolutio settigs: Start Wavelegth + Resolutio to m Resolutio/2. Resolutio Eter the umber of diodes to be averaged together as a sigle spectral data poit. To differetiate closely related spectra ad obtai greater spectral resolutio, use a small resolutio umber. Be aware, however, that a small resolutio value geerates more data poits ad therefore requires more disk space tha a large resolutio value. Fid a resolutio value just small eough to idetify spectral features. Rage: 1.2 to 24.0 m i multiples of 1.2. Samplig Rate Select the acquisitio rate i spectra per secod from the drop dow list box. For good itegratio ad quatitatio, acquire 15 to 20 spectra across a peak. Auto Exposure Check this box to eable the detector optics to calculate the optimum exposure time eeded to recharge the diodes based o the lamp eergy, the lamp spectrum ad the selected wavelegth rage. Tip: Eable Auto Exposure for most routie aalyses. Page 81

86 Iterpolate Check this box to istruct the detector to igore the sigal from the photodiode at 656 m ad to iterpolate a value from the adjacet diodes. This prevets over-saturatio at 656 m (Balmer lie for deuterium). If this box is ot checked the detector reports the sigal from the photodiode at 656 m. Disable this parameter oly if you are workig with compouds that absorb i the 656 m rage. Note: If this parameter is uchecked, the deuterium lamp high emissio lie at 656 m may cause spectral artifacts ad autoexposure errors. Exposure Time Eter the legth of time i millisecods that the photodiodes are exposed to light before they are read. This parameter is ot accessible if Auto Exposure is checked. Rage: to ms. Stop Time Eter the time, i miutes after ijectio, whe the PDA will stop scaig. This value is idepedet of the istrumet method ru time. Filter Respose Eter the respose time (i secods) for filterig acquired data. The filter is a ehaced rollig average filter applied to absorbace data from the PDA detector before the data is set to the MassLyx software. The filter reduces high-frequecy oise across the etire wavelegth rage specified for the acquisitio. High values decrease peak respose. Rage: 0 to 3. Save to Disk Check this box to save the Photo Diode Array data to the raw datafile. If this data is ot required for further processig the ucheck the box, the data is ot saved to disk thus reducig the size of the file. Waters CapLC Chael Detector Cofiguratio Pages The Chael 1 ad Chael 2 pages cotai the same iformatio. Select the page relevat to the chael required, by clickig o the tab. Figure 2.63 Chael 1 Detector Cofiguratio page Page 82

87 Output Mode Select oe of: Off o aalog output sigal. Absorbace Output represets absorbace at the wavelegth specified by the Wavelegth parameter (see below). Note: Ratio Deomiator Wavelegth ad Threshold parameters are ot accessible whe Absorbace mode is selected. Ratio Output represets the ratio of absorbaces at two wavelegths. The umerator wavelegth is specified by the Wavelegth parameter, ad the deomiator wavelegth is specified by the Ratio Deomiator Wavelegth parameter (see below). Wavelegth Eter the output wavelegth. I Ratio mode, the absorbace at the Wavelegth is used to calculate ratio i the formula: Ratio = Absorbace at Wavelegth/Absorbace at Ratio Deomiator Wavelegth Wavelegth must be withi the wavelegth rage specified by the Start Wavelegth ad Ed Wavelegth parameters o the PDA page. Rage whe Resolutio is set to 1.2: Start Wavelegth to Ed Wavelegth. Rage at all other Resolutio settigs: Start Wavelegth + (Badwidth/2) to Ed Wavelegth (Badwidth/2). Ratio Deomiator Wavelegth Eter the deomiator wavelegth (i aometers) for the aalog output chael. Ratio Deomiator Wavelegth must be withi the wavelegth rage specified by the Start Wavelegth ad Ed Wavelegth parameters i the 996 PDA page. Badwidth Eter the spectral badwidth of the aalog output chael. Rage: 1.2 to 24.0 m i multiples of 1.2. Filter Type Select the filter type (Hammig or Sigle Pole) from the drop dow list box for use o the aalog output chael. The Hammig filter is desiged to create the same degree of peak-height degradatio as the Sigle Pole filter for the same respose time, but ehaces filterig of high-frequecy oise. Filter Respose Eter the respose time i secods for the Filter Type specified above. Rage: 0 to 5 secods. Offset If required eter a offset to the aalog output chael. Rage: 0.2 to 2.0 AU. Threshold Eter a threshold above which the ratio (Wavelegth / Ratio Deomiator Wavelegth) must be to be valid data. The rage is 0.1 to 2.0 AU. Note: If o ratio is plotted (oe or both chaels are below the curret Threshold), eter a lower Threshold value. Page 83

88 Waters 515 ad 1525 Pumps The Waters 515 ad 1525 Pump pages ca be accessed by selectig Waters 515 Pump or Waters 1525 Pump from the View meu o the Ilet Editor or by pressig the toolbar butto. Waters 515/1525 Iitial Coditios Page Figure 2.64 Iitial Coditios page Solvet A Eter the ame of the solvet that will be delivered through Pump A. Solvet B Eter the ame of the solvet that will be delivered through Pump B ad eter the percetage of the solvet flow from Pump B. Flow (ml/mi) Eter the total flow rate for the solvet delivery system. Rage: 1 to 10 ml/mi. Ru Time Eter the time i miutes that the method will ru, from the poit of ijectio. Note: If you are ruig a gradiet or settig timed evets, make sure the Ru Time value is greater tha, or equal to the greatest Time value, specified o the Gradiet or Timed Evets pages. Low Pressure Eter the low pressure limit for the system. If the pressure falls below this limit, the solvet flow will stop ad the LC status light will tur red. High Pressure Eter the high pressure limit for the system. If the pressure exceeds this limit, the solvet flow will stop ad the LC status light will tur red. Page 84

89 Waters 515 If you selected a Waters 515 Pump, the Auxiliary Pump Solvet optios are displayed: Flow (ml/mi) Eter the total flow rate for the auxiliary solvet delivery system. Rage: 1 to 10 ml/mi. Name Eter the ame of the solvet that will be delivered through the auxiliary pump. Waters 515/1525 Gradiet Page Figure 2.65 Gradiet page Use the Gradiet Table to defie coditios for a gradiet ru. For each row i the Gradiet Table, you eed to defie the % compositio of up to two solvets that are to be delivered at the desired flow rate for the specified Time. Note: For a isocratic ru, set the solvet percetages, ru time ad flow, o the Iitial Coditios page. Do ot add ay rows to the Gradiet Table. To add a gradiet, eter a time ad percetage i the relevat boxes ad press the toolbar butto. Note: The first etry must have a time of 0. To delete a sigle gradiet, click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. Page 85

90 To modify a gradiet, select the required etry i the timetable. The values will the be displayed i the edit boxes to the left of the timetable, ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig timetable. Flow Eter the flow rate for the solvet delivery system. will result i a ew etry beig created i the Curve Eter the umber of the gradiet curve required. This sets the rate at which the solvet is to chage to the ew proportios ad/or flow rates. See the Waters Operator s Guide for a list of values. Waters 515/1525 Iitial Evets Page Figure 2.66 Iitial Evets page The exteral evets are triggered by four cotact closures (relays) through output termials, which are located at the back of the istrumet. Waters 515 Relays 1 ad 2 From the drop dow list box, select ON or Off to activate or deactivate the relay. Evets 1 to 4 From the drop dow list box, select ON or Off to activate or deactivate the evet. Waters 1525 Relays 1 to 4 From the drop dow list box, select ON or Off to activate or deactivate the relay. Page 86

91 Waters 515/1525 Timed Evets Page Figure 2.67 Timed Evets page Use the Evet Table to program up to 16 evets (both exteral ad iteral). Evets ca be triggered more tha oce ad multiple evets ca be triggered simultaeously. Time Eter the time (after ijectio) at which the evet starts. Evet rows are sorted automatically by time. Note: Differet evets ca be programmed to occur at the same time. Rage: 0.00 to mi. Evet Select a Evet or Relay from the drop dow list box. Switch States O Turs o a cotact closure that triggers a exteral or iteral evet. With this fuctio, the cotact closure remais closed util a Off fuctio is set. Off Turs off the cotact closure for the evet. With this fuctio, the cotact closure is broke. Page 87

92 Evet Table Operatio To add a evet, eter a time, evet, actio ad value i the relevat boxes ad press the toolbar butto. Note the first etry must have a time of 0. To delete a sigle evet, click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To modify a evet, select the required etry i the timetable. The values will the be displayed i the edit boxes, ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig will result i a ew etry beig created i the timetable. Waters 515/1525 Method Type Page Figure 2.68 Method Type page Method Type Specifies the type of method to create. Normal Method The Method Type used for stadard ijectios. Esure that the Method Type is set to Normal uless you are performig oe of the procedures listed below. Colum Coditio Method Rus solvet through the colum without ijectig samples or ruig the Evets table. Solvet is delivered usig the gradiet table specified i the Gradiet page. Equilibrate Method Delivers solvets ad maitais solvet parameters usig the values defied o the Iitial Coditios page. Ru Evet Table Check this box to ru the table of timed evets durig the method. This optio is oly active if the Normal or Colum Coditio Methods are selected. Page 88

93 Notes Page 89

94 Notes Page 90

95 CE Istrumets CE Istrumets GC8000 Gas Chromatograph Chapter 3 O a CE Istrumets GC8000, MassLyx ca cotrol the ove temperature, the ijector zoe temperatures, the valve times, the dump valve ad 4 exteral evet times. To chage GC Parameters 1. Choose Set up Ilet from the Acquisitio Cotrol Pael Istrumet meu. - or - Double click o the picture of the GC o the Acquisitio Cotrol Pael to display the GC8000 ilet editor show below. 2. Make ay chages to the parameters. Note: The ove temperature ramp ca be modified either by usig the keyboard to eter times, temperatures ad rates, or by draggig the small red hadles o the graph with the mouse. 3. Save the method usig either Save or Save As from the File meu. Figure 3.1 GC8000 Ilet Editor The time ad temperature rage of the ove temperature ramp ca be cotrolled usig the buttos displayed to the right of the ramp. Clickig o will icrease the rage show o the time axis. Clickig o will icrease the rage show o the temperature axis. Clickig o alters the display rages so that the ove temperature display fills the graph. Page 91

96 A full descriptio of all the parameters i this editor is give i the GC8000 Series Istructio Maual. Chagig Valve Evet Times Timed evets such as Purge ad Split times ca be icluded i the GC method. These are programmed usig the GC 8000 Valve Cotrol editor show below. Figure 3.2 GC8000 Valve Editor To display this dialog select Valve Timetable from the Valves meu. The evets parameters are stored to disk whe the GC parameters are saved, ot whe this dialog is closed so esure that parameters are saved before startig a acquisitio. To cotrol the GC8000 DPFC optio MassLyx ca cotrol the DPFC optio o the GC8000 for Quattro II ad Platform istrumets. 1. Select Cofiguratio from the GC8000 editor DPFC meu. Figure 3.3 DPFC Cofiguratio dialog 2. Cofigure the left ad right chaels as required. Each chael ca be set to flow, pressure or off. 3. Select Left Chael or Right Chael from the GC8000 editor DPFC meu to load either the Flow Ramp or Pressure Ramp Editor depedig how the chael is cofigured. Page 92

97 Figure 3.4 DPFC Flow Ramp Editor Figure 3.5 DPFC Pressure Ramp Editor 4. Make ay chages required ad press OK to exit ad save chages. Page 93

98 CE Istrumets AS800 Auto Ijector The CE Istrumets AS800 Auto Ijector ca be used with the CE Istrumets GC8000 gas chromatograph. The autosampler is programmed from MassLyx usig the A200S editor. It is programmed i exactly the same way as the A200S system described above. A full descriptio of all the parameters i this editor is give i the AS800 Autosampler Istructio Maual. Figure 3.6 AS800 Auto Ijector Editor Page 94

99 Notes Page 95

100 Notes Page 96

101 Gilso Systems Gilso Autosamplers Chapter 4 Itroductio Supported Models The Gilso Software ca be used to cotrol ay of these models: Gilso 215 Gilso 232XL Gilso 231XL Gilso 233XL Gilso 222XL The Gilso 232XL ad 233XL also require the Gilso 402 Dilutor. We also support the Gilso 401C Dilutor although this has ow bee discotiued by Gilso. The Gilso 215 has a dilutor built i, but it does require a Gilso 819 Valve Actuator. Settig up The first time MassLyx is ru with a Gilso Autosampler it eeds to kow which autosampler is beig used. The followig dialog is displayed. Figure 4.1 Gilso Defaults dialog Select the Autosampler, Dilutor ad Ijector istalled ad press OK. Note: This oly eeds to be doe the first time you use MassLyx or if the type of autosampler is chaged. Page 97

102 The Gilso Toolbar The Gilso toolbar has three extra buttos o it, which are: Click To Prime the Dilutor. Gilso Cofiguratio Pages Geerate Custom Racks. These pages cotai iformatio that is used to cofigure the autosampler. To access them press the Ilet Editor View meu. butto or select Gilso AutoSampler from the The Gilso Task List This page is used to build up a set of tasks ito a method that is the used to perform the ijectio. The available tasks are cotaied i the Task drop dow list box ad the parameters displayed will deped o which task is selected. Figure 4.2 Task List page Page 98

103 Addig ad Deletig Tasks (Gilso) To add a task select it from the Task drop dow list box, set the parameters ad press the add butto. The task will be added to the ed of the list. To delete a sigle task, select it from the Task List ad press the delete butto. To clear all of the tasks i the Task List press the clear all butto. Modifyig Tasks (Gilso) To modify a task select it from the Task List, chage the required parameters ad press the add butto. Movig ad Copyig Tasks (Gilso) To move a task, select it with the mouse, hold the mouse butto dow ad move the task to the required positio. To copy a task follow the procedure for movig tasks but hold the CTRL key dow whe movig the mouse. The Idividual Tasks (Gilso) This sectio cotais tables listig the idividual parameters for each task. Iject Partial Loop This is the mai task that is used to iject. The Ijectio volume is etered i the sample list. Parameter Aspiratio Flow Rate Displacemet Flow Valve Flush Volume. Descriptio This is the flow rate at which the sample is draw ito the eedle. This is the flow rate at which the sample is ijected ito the loop. This is the volume of solvet that is flushed through the valve after the ijectio. It is displaced at the Displacemet flow. Table 4.1 Iject partial loop parameters Page 99

104 Detailed below is a list of the steps that occur durig the iject partial loop task. Steps: 1. Move to vial 2. Aspirate air gap if required 3. Move Zarm dow to vial depth 4. Aspirate ijectio volume + ijectio flush volume 5. Move Zarm back to travel height 6. Move to ijectio port 7. Move to ijectio depth 8. Switch valve to iject positio 9. Dispese ijectio flush volume 10. Switch valve to load 11. Dispese ijectio volume 12. Switch valve to iject 13. Pulse output cotact 14. Dispese air gap 15. Rise valve if a flush volume has bee specified Note: The Left ad right valve become eabled whe usig a Gilso 233XL. Rise Ijectio Port This task is used to rise the ijectio port with solvet from the reservoir. Parameter Risig Volume Displacemet Flow Descriptio This is the volume of solvet that is to be rised through the eedle. This is the flow rate at which the solvet is ijected ito the valve. Table 4.2 Rise Ijectio Port parameters Note: The Left ad right valve become eabled whe usig a Gilso 233XL. Page 100

105 Rise Iside Needle This task is used to rise the iside of the eedle with solvet from the reservoir. Parameter Risig Volume Displacemet Flow Rise Statio Descriptio This is the volume of solvet that is to be rised through the eedle. This is the flow rate at which the solvet is rised through the eedle. This is the rise statio at which you would like the rise to take place. If this is set to auto the the earest rise statio is chose. Table 4.3 Rise Iside Needle parameters Rise Outside Needle This task is used to rise the outside of the eedle with solvet from the reservoir. Parameter Risig Volume Displacemet Flow Depth Rise Statio Descriptio This is the volume of solvet that is to be rised through the eedle. This is the flow rate at which the solvet is rised through the eedle. This is set to the depth that the eedle should move to for risig. This is the rise statio at which you would like the rise to take place. If this is set to auto the the earest rise statio is chose. Table 4.4 Rise Outside Needle parameters Because this task dispeses the solvet through the iside of the eedle it also acts as a rise iside eedle. Note: Risig the Outside of the eedle is ot available o the Gilso215 Set Electrical Cotact This task is used whe you would like to set oe of the output cotacts. Parameter Cotact Number State Descriptio This is the umber of the cotact that you would like to set. This is the state that you wat to set. Ope, Close or Pulse. Table 4.5 Set Electrical Cotact parameters Page 101

106 Wait For Cotact This task is used whe you eed to wait for a cotact state. Parameter Cotact Number State Descriptio This is the umber of the cotact that you would like to set. This is the state that you wat to set. Ope, Close or Pulse. Table 4.6 Wait For cotact parameters Wait For Time This task is used if you eed the machie to wait for a specified time. Parameter Time Descriptio The time that you wish to wait for. Table 4.7 Wait For Time parameters Gilso Dilutor Cofiguratio Page This page is used to set parameters specific to the Dilutor. Figure 4.3 Dilutor Cofiguratio page Syrige Volumes The size of the curretly istalled Left ad Right syriges. Reservoir Aspiratio Flow This is the flow rate at which solvet is draw from the reservoir (the default value is usually sufficiet). Prime Displacemet Flow This is the flow rate at which the solvet is displaced durig the prime dilutor (the default value is usually sufficiet). Page 102

107 Ilet Tubig volume This is the volume of the tubig betwee the ijectio port ad the rheodye valve. It is the volume of air used to push the sample through ito the ijectio loop. Sample Flush volume This is the amout of sample that is draw with the ijectio valve. This amout is the ijected before the valve switches to fill the tube betwee the valve ad the ijectio port. Ay excess will go to waste. Note: Whe usig the Gilso 402 Dilutor, the Left ad Right syrige radio buttos are eabled allowig you to use the left ad right syriges. This is ot yet available. Gilso Sampler Cofiguratio Page This page is used to set parameters specific to the Sampler. Figure 4.4 Sampler Cofiguratio page Curret Layout Select the required layout from the drop dow list box. A picture of the selected bed layout is displayed below the ame. Rise Statios Preset Check the boxes if the rise statios are preset. Left ad right are as you look at the machie. Note: Whe usig the Gilso 215 the right rise statio will be disabled. Page 103

108 Gilso Multi-Ijector Parameters Page This page is used to set parameters specific to the Gilso Multi-Ijector System. This is a autosampler device that ijects up to 8 samples simultaeously. The Multi-Ijector itself cosists of a Gilso 215 autosampler i cojuctio with a Gilso 889 multi-valve ijector. The device has eight eedles coected to the robot arm ad a syrige for each eedle. The geometry of the autosampler is desiged so that the 8 eedles will eter 8 separate wells of a microtitre plate. The 8 samples are picked up at the same time ad the deposited ito 8 separate ijectio loops. The valves coected to these loops ca the be cotrolled idividually so that each of the samples ca be set to the mass spectrometer at ay specified times. To eable these parameters the followig cofiguratio must be defied. 1. Select Advaced Cofiguratio from the Gilso Sampler meu. 2. O the Hardware tab select Gilso 215 from the Sampler drop dow list box. 3. O the Hardware tab select Gilso 889 from the Valve drop dow list box. 4. O the Hardware tab select All samples to same file from the Multi-Ijector Mode drop dow list box. 5. O the Hardware tab check the Eable Iject Ahead box. 6. Press OK. Figure 4.5 Multi-Ijector Parameters page Sca Time per Sample This defies the aalysis time spet o each sample. Delay This is the time for the sample to travel from the ijectio port to the mass spectrometer. Legth This is the amout (i secods) of the chromatogram that is used to create the data files for each of the samples. Page 104

109 Gilso Multi-Ijector Processig A complete row of 8 samples are collected simultaeously ad placed i the ijectio ports. The ijectio valve for the first sample is switched to iject ad a cotact closure is used to sigal the mass spectrometer to iitiate scaig. Each subsequet sample is the ijected every T secods, where T is the Sca Time Per Sample defied o the multi-ijector parameters page. Oce all 8 samples have bee ijected the ext set of samples are loaded i the ijectio loops. The first of these samples is ijected after a time T from the last sample of the previous row. The scaig time defied i the Mass Spectrometer method should be log eough so that all the samples ca be ijected ad scaed. This time has to be calculated by the user. Data for all samples is writte to oe file, the BatchDataFile, see Figure 4.6. Each peak withi this chromatogram relates to a separate sample. Default file BATCH_Default % Sca ES+ TIC 4.26e Figure 4.6 Multi-Ijector Batch File Time The file ame of the BatchDataFile is either based o the sample list ame or the OpeLyx Job ID. A prefix of BATCH_ is added to this ame so that it ca be recogised as a BatchDataFile. E.g. for a sample list called Test.SPL, the ame of the BatchDataFile will be BATCH_Test.RAW. Oce the scaig for the BatchDataFile is complete ChroSplit.exe is used to create idividual data files for each sample i the batch. ChroSplit.exe should be defied i the Acquire Process colum for the first sample i the sample list or o the OpeLyx Setup Acquisitio Process page. ChroSplit reads the BatchDataFile.RAW ad splits it ito idividual files based o the values defied o the Multi-Ijector Parameters page. It igores the first part of the file that is the Delay. The rest of the file is split ito Sca Time per Sample sectios ad the scas acquired from the start of each of these sectios for the defied Legth are copied ito the idividual data files. Figure 4.7 Multi-Ijector Batch File Page 105

110 Some of the scas betwee ay two samples will cotai some mixig of samples (e.g. Regio B i Figure 4.7 ) or ot cotai data relatig to either sample (e.g. Regio A i Figure 4.7 ). The data i these regios is ot required, therefore oly data i Legth regio is copied. Oce all data files have bee created by ChroSplit the acquisitio will proceed with ay processig defied i the Process colum for the first sample i the batch. For all subsequet samples i the batch o data is acquired oly the processes are iitiated. ChroSplit curretly writes two data files per sample. The first has a re-ormalised retetio time so that the retetio time of the first sca for each sample is set to 0. This allows aalysis of the masses to proceed via loop ijectio, which simply searches for masses cotaied withi the sca at a user-defied retetio time. The other file writte retais its origial retetio time ad is used to assess the accuracy of the ChroSplit procedure. This secod data file has the prefix CHECK_ added to the.raw file ame. Figure 4.8 The results of usig ChroSpilt.exe. Upper shows the Batch file, middle the re-ormalised files ad lower the check files Page 106

111 Problems with ChroSplit If ChroSplit fails to copy the BatchDataFile scas correctly (due to a icorrect delay beig defied or experimetal faults, e.g. a chage i pumpig speed) the the delay ad legth parameters ca be chaged ad the BatchDataFile reprocessed. To do this the values i the timefile.tfl eed to be chaged. This file cotais the time each sample reaches the mass spectrometer ad is stored i the BatchDataFile.RAW directory. Also writte to this file are the sample umber, sample locatio for each sample ad the multi-ijector legth ad delay parameters. [timefile] MasterRawFile=Batch_xxxxx Delay=4.5 Legth=15 [Sample] [Locatio] [Ijectio Time] [Sample ID] 1 "1,A" "Not used" 2 "1,B" "Not used" 3 "1,C" "Not used" 4 "1,D" "Not used" Figure 4.9 Example timefile.tfl 1. Ope the timefile.tfl usig a text editor, chage the Delay or Legth as required ad save the file. 2. Ope the Sample List ad chage the Process for the first sample to ChroSplit.exe. 3. Press the butto to display the Start Sample List Ru dialog. 4. Esure that oly the Auto Process Samples box is checked, the Ru From Sample = 1 ad the Ru To Sample = 1, the press OK. 5. This will split the BatchDataFile ito the idividual *.RAW files. 6. O the Sample List chage the Process back to the origial processame ad repeat step Esure that oly the Auto Process Samples box is checked, the Ru From Sample = 1 ad the Ru To Sample = last sample i the list, the press OK. The data file for each sample will be recreated overwritig the previous files. See the OpeLyx Users Guide, Itroductio chapter for details o importig OpeLyx Batch files ito the Sample List. Page 107

112 Gilso Advaced Optios Gilso Tray Optios To display the Tray Optios dialog, select Optios from the Gilso Tools meu o the Ilet Editor. Figure 4.10 Tray Optios dialog Colours Select the colour to display the Selected vial, No selected vial ad Backgroud from the appropriate drop dow list box. Display Gridreferece o tray Check this box to display the grid referece o the tray. Gridreferece text From the drop dow list box, select the colour i which you wat the grid referece text to be displayed. Page 108

113 Gilso Plate Geerator To display the Custom Rack Geerator dialog, select Plate Geerator from the Gilso Tools meu o the Ilet Editor dialog or press the butto. Figure 4.11 Custom Rack Geerator Rack Name The ame of the plate that is curretly beig edited. Origi The corer of the rack that the vial grid referecig starts from. Rows The umber of vials i a row ad the distace betwee each ceter. Colums The umber of vials i a colum ad the distace betwee each ceter. Vial Diameter does ot affect ay parameters apart from how the tray looks i the Rack Geerator. Depth affects how deep the eedle travels ito each vial whe samplig. Decreasig the depth value will make the eedle travel dow further ito the vial. This cotrol is very importat. A icorrect settig could sed the eedle through the bed ad bed it. Grid Referece Allows the user to select the way that the vial rows ad colums are refereced, e.g. whether the rows are alphabetical or umerical. Referecig This has three optios XY which refereces the vials A1, B1 etc. Sequetial Discotiuous which umbers the vials 1, 2, 3 across a row, left to right, ad the starts the ext row from the left agai. Sequetial Cotiuous which umbers the vials 1, 2, 3 across a row, left to right, the cotiues umber the ext row, right to left etc. If the Gilso autosampler is used with OpeLyx the the vial referecig must be set to either sequetial cotiuous or sequetial discotiuous. Priority Check the Horizotal First box if samples are to be acquired horizotally across the plate. Page 109

114 If Referecig = X,Y, Horizotal = Letter, Vertical = Number ad Horizotal Priority is checked, this will result i samples beig acquired i the order A1, A2, A3. If the Horizotal Priority box is ot checked samples will be acquired i the order 1A, 1B, 1C etc. If Referecig = sequetial cotiuous or discotiuous ad Horizotal Priority is checked, this will result i samples beig acquired from row 1 the row 2. If the Horizotal Priority box is ot checked samples will be acquired from colum 1 the colum 2 etc. Offsets Allows alterate vial rows or colums to be offset. Plate Size The size of the plate to its outside edges. Top Left Vial Offset The measuremet to the ceter of the first vial from the top left corer of the plate. Creatig ad Deletig Plates (Gilso) To create a ew rack press the butto. A ew default rack is displayed, chage the Rack Name, eter the appropriate values ad press the save butto or select Save Curret Rack from the Rack meu. New racks are saved to the MassLyx Plates directory. To copy a custom rack, page through the list of saved custom racks usig the ad toolbar buttos. The Previous Rack ad Next Rack optios o the Rack meu perform the same operatio. Whe the required rack is displayed chage the Rack Name, eter the appropriate values ad press the save butto or select Save Curret Rack from the Rack meu. New racks are saved to the MassLyx Plates directory. To delete a custom rack select the rack to delete, by typig the ame i the Rack Name box or by pagig through as above, ad press the delete choose Delete Curret Rack from the Rack meu. butto or Note: All of the spacigs ad the vial sectio are stored i 0.1 mm uits. Note: Whe defiig a custom plate for use with a multi-ijector the plate is required to be compatible with the positio of the 8 eedles of the autosampler. The Plate must have eight colums. The positio of the vials should allow all eight eedles to eter a separate vial. There should be o odd or eve offsets for ay of the vial positios. Default Plate Settigs (Gilso) Selectig Default Settigs for New Rack from the Tools meu displays the Default Settigs dialog. This dialog allows the default settigs used whe creatig a ew rack to be defied. Field descriptios are the same as above. Page 110

115 Rotatig ad Scalig Plates (Gilso) Selectig Rotate Rack from the View meu will rotate a rack by 90 degrees. For example: Selectig Scale Rack from the View meu displays the followig dialog. Figure 4.12 Scale Rack dialog Move the slider or eter a ew value to chage the size of the rack as displayed i the Plate Geerator dialog. Page 111

116 The Gilso Bed Layout Editor To display the Bed Layout Editor dialog, select Bed Layout Editor from the Gilso Tools meu o the Ilet Editor dialog. Figure 4.13 Bed Layout Dialog To Create a New Bed Layout (Gilso) 1. Highlight a bed layout similar to the oe you wat to create ad press the butto to create a ew layout. The layout appears i the Bed Layouts list as the same ame with a 1 at the ed, e.g. Code 201 Bed Layout1. 2. To chage the ame of the layout, type the ew ame ito the Bed Layouts text box ad press the list box. butto. The ame is updated i the Bed Layouts New bed layouts are saved to the MassLyx Racks directory. To Delete a Bed Layout (Gilso) 1. Highlight a bed layout you wish to delete ad press the butto. A dialog box will ask you to cofirm the deletio. Press the OK butto to delete the bed layout. Page 112

117 Modifyig the Number of Rows ad Colums (Gilso) To chage the umber of rows i the curret colum, type the ew umber ito the Rows box ad press the butto. To apped a ew colum, press the butto. To delete the curret colum press the butto. To isert a colum, click o the colum before which you wat to isert ad press the butto. Note: The colum iserted will have the same umber of rows as the colum highlighted. Modify Bed Layout (Gilso) If the plate positio or type eeds chagig select the Modify Bed Layout tab. Figure 4.14 Modify Bed Layout Dialog Click o oe of the code plates to display the Plate Positio ad Type dialog. Page 113

118 Figure 4.15 Plate Positio ad Type dialog This dialog allows you to select a ew plate from a list of possible optios, ad chage its actual positio o the bed. Measuremets for plate positios are always take from the top left corer of each plate. The X value is the measuremet from the curretly selected plate to the plate immediately to the left. The Y value is the measuremet from the curretly selected plate to the plate immediately above. If there is o plate, to the left or above, the measuremets are take from the Home positio, which is where the eedle sits whe ot i use. Rotated Check this box if the plate is rotated. OpeLyx plate logi If this box is checked ad Use curret MassLyx autosampler bed layout is checked i the OpeLyx Maager program, the the plate at this positio ca oly be used for plate logi o the OpeLyx Logi program. Page 114

119 Adjustig The Arm Height o a Gilso 215 The first time the Gilso software cofiguratio pages are accessed the followig dialog will be displayed. Type i the Needle Height that you have set the Gilso to ad press OK. Figure 4.16 Ijectio Parameters dialog. Page 115

120 Other Advaced Optios (Gilso) The advaced optios dialogs ca be accessed by choosig Advaced Cofiguratio from the Gilso Sampler meu optio o the Gilso AutoSampler dialog. The parameters o these pages will be set up at istallatio ad should ot eed chagig. Figure 4.17 Hardware Cofiguratio Dialog This page defies the type of Gilso AutoSampler, Dilutor ad Valve used. It also shows the state of the iitial electrical cotacts to the AutoSampler ad pump, the pulse width ad eedle parameters. Page 116

121 Figure 4.18 Positios Dialog This page defies the iteger positio from the Home positio of the eedle, i.e. the positio of the eedle whe ot i use. It also defies the distaces the eedle eeds to travel to the rise statios. Figure 4.19 Commuicatios Dialog This page defies the serial lie commuicatio betwee the Gilso ad the PC. Page 117

122 Gilso Pump The Gilso Pump pages ca be accessed by selectig Gilso Pump from the View meu o the Ilet Editor or by pressig the toolbar butto. Gilso Solvets ad Flows Page Figure 4.20 Solvets ad Flows page Solvets Up to four solvets will be displayed depedig upo the system cofiguratio. The total value of all the solvet percetages added together must ot exceed 100%. Pump A This is the remaider percetage after the solvet percetages have bee set for the other pumps. Pump B, C, D These ca either be eabled or disabled by checkig the box ext to the idividual pumps. The values ca be set to the required percetage of flow delivery. Solvet Name Type i the solvet ame. Refill Time This is the time required for the pisto retur stroke. Normally it is set to the lowest value (125ms). If cavitatio or degassig occurs, the a higher value must be used. The miimum value is 125ms ad the maximum 1000ms. Compressibility This is used to calculate a flow rate compesatio for the compressibility of the solvet. See the Gilso User Guide for suitable values. Ru Time This is the legth of time, i miutes, the pump should ru for. Flow Rate This is the total flow rate of the solvet chaels accordig to how you have cofigured the istrumet. Page 118

123 Gilso Gradiet Timetable Page Figure 4.21 Gradiet Timetable page This page allows a gradiet to be etered ad edited. To operate i isocratic mode esure that the timetable is empty. To add a gradiet, type i a time, the required percetages ad the flow rate, i the relevat boxes ad press the time of 0. toolbar butto. Note: The first etry must have a To delete a sigle gradiet click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To modify a gradiet, select the required etry i the timetable. The values will the be displayed i the edit boxes above the timetable, ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig will result i a ew etry beig created i the timetable. Page 119

124 Gilso Evets Timetable Page Figure 4.22 Evets Timetable page To add a evet, eter a time, select a evet from drop dow box ad press the toolbar butto. To delete a sigle evet, click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. To modify a evet, select the required etry i the timetable. The values will the be displayed i the edit boxes above the timetable, ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig will result i a ew etry beig created i the timetable. Page 120

125 Gilso Pressures Page Pump Cofiguratio Figure 4.23 Pressures page To set pressure limits check the Maometric Module box ad eter a Miimum ad Maximum pressure. The maximum pressure limit is determied by the smallest pumphead size. To chage the umber of pumps used select Pump Cofiguratio from the Gilso Pump meu. Figure 4.24 Pump Cofiguratio dialog Select the umber of pumps, the pump type, pump head ad the GSIOC ID (Gilso Serial Iput Output Chael) required. Page 121

126 Page 122

127 Notes Page 123

128 Notes Page 124

129 Hewlett Packard Systems Hewlett Packard 5890 Gas Chromatograph Chapter 5 Both the HP5890 Series I ad Series II ca be cotrolled by MassLyx. For the Series II istrumet it is ecessary to cofigure the GC to respod to Series I commads by settig a jumper i the GC. Your istallatio egieer will have doe this. Before startig to use the GC the software must be cofigured to reflect the GC equipmet i use e.g. the umber of ijectors ad detectors etc. To chage GC Parameters (Hewlett Packard 5890 Gas Chromatograph) 1. Choose Set up Ilet from the Acquisitio Cotrol Pael Istrumet meu or Double click o the picture of the GC o the Acquisitio Cotrol Pael to brig up the HP5890 ilet editor show below. 2. Make ay chages to the parameters. Note: The ove temperature ramp ca be modified either by usig the keyboard to type i times, temperatures ad rates, or by draggig the small red hadles o the graph itself usig the mouse. 3. Save the method usig either Save or Save As from the File meu. Figure 5.1 HP5890 Editor The time ad temperature rage of the ove temperature ramp ca be cotrolled usig the buttos that appear to the right of the ramp. Clickig o will icrease the rage show o the time axis, clickig o will icrease the rage show o the temperature axis. Clickig o alters the display rages so that the ove temperature display fills the graph. A full descriptio of all the parameters i this editor is give i the HP5890 Gas Chromatograph Referece Maual. Page 125

130 To Chage GC Cofiguratio (Hewlett Packard 5890 Gas Chromatograph) 1. Choose Cofiguratio from the HP5890 Edit meu to display the cofiguratio editor show below. 2. Make ay chages to the parameters. 3. Press OK. The parameters will be saved with the GC method whe either Save or Save As is selected from the HP5890 File meu. Buttos i the HP5890 Edit meu that are ot appropriate for the selected cofiguratio will be grayed out. Figure 5.2 HP5890 Cofiguratio Editor The mai HP5890 editor is used to set up the GC ove temperature program ad to cotrol the dump valve ad cryogeic coolig optios if fitted. To chage Purge Valve Times (Hewlett Packard 5890 Gas Chromatograph) Figure 5.3 HP5890 Purge Valve Editor Page 126

131 Hewlett Packard 7673A Auto Ijector The HP 7673A auto ijector ca oly be used with the HP 5890 gas chromatograph. Figure 5.4 HP7673A Auto Ijector Editor Note: This dialog oly applies if the Dice optio is selected at setup, if the Dice optio is ot selected a No parameters to set message is displayed. To Chage Autosampler parameters (Hewlett Packard 7673A Auto Ijector) 1. Choose Set up Auto Ijector from the Acquisitio Cotrol Pael Istrumet meu or Double click o the picture of the auto ijector o the Acquisitio Cotrol Pael to display the HP7673A editor show above. 2. Make ay chages to the parameters. 3. Save the method usig either Save or Save As from the File meu. A full descriptio of all the parameters i this editor is give i the HP7673A Automatic Sampler Operatig Maual. A autosampler will usually be used with the multiple sample acquisitio page where further iformatio such as bottle umber will be etered. Startig a acquisitio with a autosampler will be covered i the ext sectio Page 127

132 Hewlett Packard HPLC Systems The HP1050, HP1090 ad HP1100 HPLC systems ca be cotrolled from MassLyx. All three photo diode array (PDA) ad UV detectors are supported. The software ca be used to cotrol the pump durig istrumet tuig or acquisitio ad ca be used to provide multi-sample acquisitios. Both isocratic ad gradiet modes of operatio are supported. The HP1050 is described i the examples below. Differeces for the HP1090 ad HP1100 are also described. Hewlett Packard Sampler Cofiguratio Page Select the HP1050 AutoSampler, HP1090 AutoSampler or HP1100 AutoSampler from the View meu or press the toolbar butto. Figure 5.5 HP1050 Sampler Iitial Coditios Widow Ijectio Volume This is the volume i microlitres to iject. Note: If you are ruig from the Sample List the ijectio volume i the sample list etry overrides the settig defied here. Draw Speed This determies the rate i microlitres per miute at which sample is extracted ito the autosampler eedle. This should be set accordig to the viscosity of your sample. Eject Speed This is the speed i microlitres per miute at which sample is ejected from the eedle o ijectio. Agai set this accordig to the viscosity of the sample. Cosult your HP documetatio for further iformatio. Note: This facility is oly available with the HP1050 ad HP1100 ad will ot be visible o a HP1090 system. Page 128

133 Draw Positio This is a offset value i mm from positio 0 ad determies how far the eedle is iserted ito your sample. Cosult your HP documetatio for further iformatio. Agai this facility is oly available with the HP1050 ad HP1100 ad will ot be visible o a HP1090 system. Stop Time This value is set i miutes to be the time that the autosampler method will ru after ijectio. This does ot apply to the HP1090 sice this has a i-built autosampler. Exteral Vial Tray If a exteral vial tray is used check this box. Vial umber The vial to iject from. Note: If a multisample acquisitio is beig ru from the MassLyx Sample List, the Bottle # etry i the sample list overrides the value defied i the Vial Number box. Syrige Size Set this to the size of syrige fitted o the HP1090 LC System. This parameter applies to the HP1090 oly ad will ot be visible i a HP1050 or HP1100 system. Thermostat O If the autosampler is fitted with a sample heater the this box will be eabled. Check it to use the sample heater. This parameter applies to the HP1100 oly ad will ot be visible i a HP1050 or HP1090 system. Sample Temperature Eter the temperature to heat the sample to. This parameter applies to the HP1100 oly ad will ot be visible i a HP1050 or HP1090 system. Hewlett Packard Sampler Exteral Evets Page Exteral evets allow cotrol of the exteral cotacts foud o the HP1050 ad HP1090 LC systems. I additio HP1090 colum switchig ca also be cotrolled. For the HP1100 a separate cotact board must be istalled i the pump i order to use this fuctioality. Figure 5.6 HP1050 AutoSampler Exteral Evets page Page 129

134 The cotacts ca be set to operate uder timed cotrol durig a method ru, as well as havig their iitial states set. The cotacts will be i the iitial state before a method ru ad will retur to this state after a method has completed. To add a evet, eter a Time, select a Evet Type from the drop dow list box ad press the toolbar butto. To delete a sigle evet click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To chage the State of a cotact, double click o the time etry i the list to toggle betwee states. Time The time i miutes at which the cotact evet should occur. Evet The cotact evet to be performed. State The state determies whether the cotact is to be opeed or closed. Hewlett Packard Pump Iitial Coditios Page Select the HP1050 Pump, HP1090 Pump or HP1100 Pump from the View meu or press the toolbar butto. Figure 5.7 HP1050 Pump Iitial Coditios page Solvets Up to four solvets will be displayed depedig upo the system cofiguratio. The total value of all the solvet percetages added together must ot exceed 100%. Pump A This is the remaider percetage after the solvet percetages have bee set for the other pumps. Page 130

135 Pump B, C, D These ca either be eabled or disabled by checkig the box ext to the idividual pumps. The values ca be set to the required percetage of flow delivery. Flow This is the total flow rate of the solvet chaels accordig to how you have cofigured the istrumet. Pressures These set the upper ad lower limits of the pressure withi the solvet delivery system (SDS) if the pressure falls outside of this rage the SDS switches off. Colum Heater If the istrumet has a ove preset the the colum temperature ca be set to a specified temperature i degrees cetigrade. Check the Colum Temperature box ad eter a temperature. If the software has bee cofigured to operate without a colum ove the these boxes will be grayed out. Note: For the HP1100 a temperature should be etered i both the Left ad Right boxes. Stop Time This value is set to the time i miutes that the method will ru from the poit of ijectio. Hewlett Packard Pump Gradiet Timetable Page Figure 5.8 HP1050 Pump Gradiet page This page allows a gradiet to be etered ad edited. To operate i isocratic mode esure the timetable is empty. To add a gradiet, type i a time ad percetage i the relevat boxes ad press the toolbar butto. Note: The first etry must have a time of 0. To delete a sigle gradiet, click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. Page 131

136 To modify a gradiet, select the required etry i the timetable. The values will the be displayed i the edit boxes above the timetable, ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig will result i a ew etry beig created i the timetable. The gradiet parameters that you ca set are as below. Note: The umber of solvet percetages, which appear i the dialog, depeds o which type of gradiet was selected i the LC Cofiguratio Widow. Time The time at which you wish the followig parameters to be attaied durig a method ru. %B The percetage of solvet B you wish to attai at the give time. %C The percetage of solvet C you wish to attai at the give time. %D The percetage of solvet D you wish to attai at the give time. Flow The required flow i ml/mi that you wish to attai at the give time. Pressure This is oly available o the HP1050 ad HP1100 ad allows the limitig high pressure (i bars) to be reset at the give time. Hewlett Packard Pump Exteral Evets Page Exteral evets allow cotrol of the exteral cotacts foud o the HP1050 ad HP1090 LC systems. I additio HP1090 colum switchig ca also be cotrolled. For the HP1100 you must have the separate cotact board istalled i your pump i order to use this fuctioality. Figure 5.9 HP1050 Pump Exteral Evets page The cotacts ca be set to operate uder timed cotrol durig a method ru, as well as havig their iitial states set. The cotacts will be i the iitial state before a method ru ad will retur to this state after a method has completed. Page 132

137 To add a evet, type i a Time ad select a Evet Type from the drop dow list box ad press the toolbar butto. To delete a sigle evet, click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To chage the State of a cotact, double click o the time etry i the list to toggle betwee states. Time The time i miutes at which you would like the cotact evet to occur. Evet Type This determies the cotact evet to be displayed i the list of evets. Oce displayed, the evets ca be added, deleted or chaged as described above. State The state determies whether the cotact is to be opeed or closed. To Set-up the Diode Array Detector (Hewlett Packard) Select the HP1050 PDA Detector from the View meu or press the butto. toolbar Figure 5.10 HP1050 DAD Spectrum Widow Rage Eter the miimum (Rage) ad maximum (To) wavelegths i aometers over which diode array spectral data will be acquired. Peak Width Determies the rate at which the data is acquired. There are approximately eight spectra per peak so a peak width of 0.1 miutes meas eight spectra will be acquired every 6 secods. Iterval This determies the umber of spectral data poits acquired. For example a iterval of 4 aometers meas data poits will be acquired at the lower wavelegth, the lower wavelegth plus 4m ad so o. Page 133

138 Autobalace Check this box to zero the base lie of the diode array detector before each aalysis. Use Pump Stop Time Check this box use the Stop Time defied o the Pump Iitial Coditios page (see page 130). Stop Time This optio is ot eabled if the Use Pump Stop Time box is checked. It determies the time i miutes the diode array method will ru. Data will be acquired for this amout of time. Save Data to Disk Check this box to store the diode array data to disk. If you do ot wish to save the diode array data to disk you should ucheck this box. HP1100 DAD The HP1100 DAD dialog has Pre Autobalace ad Post Autobalace i place of Autobalace. Check the relevat box to zero the baselie of the diode array detector before or after each aalysis. Hewlett Packard 1050 UV Detector Select HP1050 UV Detector from the View meu or press the toolbar butto. Figure 5.11 HP1050 UV Detector Widow Respose Select 250, 1000 or 4000 msec from the drop dow list box. Wavelegth Eter the wavelegth i aometers to be moitored. Page 134

139 Hewlett Packard 1090 UV Detector Select HP1090 UV Detector from the View meu or press the toolbar butto. Figure 5.12 HP1090 UV Detector Widow Respose Select oe of the values from the drop dow list. Atteuatio Select oe of the values from the drop dow list. Zero Percet Icrease the value to icrease the baselie. Filter Positio Set this to the umber of the filter required. Lamp Curret Set to Normal or High. Page 135

140 Hewlett Packard 1100 UV Detector Select HP1100 UV Detector from the View meu or press the toolbar butto. Figure 5.13 HP1100 UV Detector Widow Wavelegth Set to the wavelegth i aometers to be moitored. Respose Select oe of the values from the drop dow list. Zero Offset Icrease the value to icrease the baselie. Atteuatio Select oe of the values from the drop dow list. Page 136

141 The HP6890 System Status Page The System Status page displays iformatio about the state of the machie beig cotrolled. This page ca be accessed from the Ilet Editor by selectig Status from the View meu or by pressig the toolbar butto. Figure 5.14 System Status page Idicators The Ruig ad Ijector Cycle idicators at the left of the scree give iformatio o the curret status of the GC system. The OK ad Ready Idicators become illumiated i red if the GC System has a error. Click o the red idicators to display more iformatio o the cause of the error. Ru Time This displays how log the method has bee ruig. Flow Rate This is the curret flow rate as retured by the istrumet. The curret value will be the setpoit whe usig a flow based mode or the actual flow if i a pressure based mode. Pressure This displays the curret pressure i the istrumet. The curret value will be the setpoit whe usig a pressure based mode or the actual pressure if i a flow based mode. Temperature This displays the curret ove temperature of the GC. Page 137

142 HP6890 Sampler Cofiguratio Page Select the HP6890 AutoSampler from the View meu or press the butto. toolbar Figure 5.15 HP6890 Status ad Coectivity page Fault A red light idicates that the Autosampler has developed a fault. Ruig A gree light idicates that the Autosampler is active. The Autosampler is active if it is movig a vial, risig or ijectig. Ready A gree light idicates that the Autosampler is ready to start processig aother vial. Autosampler Has Tray Check this box if the Autosampler has a tray. Autosampler Has Barcode Reader Check this box if the Autosampler has a barcode reader. Sample Number Eter the vial umber that the ijectio will be take from, whe the start butto is pressed. Start Press this butto to start a autosampler (ad hece GC) ru with the curretly stored method. Stop Press this butto to stop the Autosampler. It will ot stop a GC ru. Page 138

143 HP6890 Ijectio Params Page Figure 5.16 HP6890 Ijectio Parameters page Ijectio Volume Eter the ijectio volume to be used for a ru started usig the Start butto o the Status ad Coectivity page. Note: If the acquisitio is started from the Sample List the this value will be overridde by the sample list ijectio volume. Tray Temperature This is used for record purposes oly ad is the tray temperature at which this method is ormally used. Solvet Washes These allow a rise strategy for each sample to be set up. Eter the umber of washes of each type i the required boxes. HP6890 Syrige Params Page Figure 5.17 HP6890 Syrige Parameters page Page 139

144 Sample Pumps Eter the umber of times the syrige will draw i liquid i order to fill it. Viscosity Delay Select the legth of time that the eedle will stay i the vial to esure that all the sample has bee draw ito the syrige, from the drop dow list box. Slow Pluger For viscous liquids select YES from the drop dow list box otherwise select NO. Allow Samplig Offset Check this box to eable the Samplig Offset. Samplig Offset Eter the distace from the bottom of the vial (i millimeters) that the ijectio will be take from. This is to allow samples to be take from differet parts of a multi-phased sample. Note: This box will bot be eabled uless the Allow Samplig Offset box is checked. Pre Dwell Time ad Post Dwell Time If, for example the sample eeds heatig before ijectio, the syrige eedle ca be held i the GC ilet for Pre Dwell Time ad/or Post Dwell Time. Eter the legth of time i the relevat box. To Set-up the Pump (HP6890) Select the HP6890 Pump from the View meu or press the toolbar butto. HP6890 Status Page Figure 5.18 HP6890 GC Status page Prep Ru This will be yellow whe the GC is i the Prep Ru state. This occurs whe the GC is tryig to equilibrate before a automatic ijectio start. Ruig This will be yellow whe the GC is i the Ruig state. This occurs whe the GC has started its temperature ad/or pressure profiles for a ru. Post Ru This will be yellow whe the GC is i the Post Ru state. Page 140

145 Ready This will be gree whe the GC is equilibrated ad ready to start a ru. Error This will be red whe the GC is i a Error state or there is a commuicatio problem betwee the GC ad the Host (local PC). The Ru Time, Curret temperature, Curret Pressure ad Curret Flow are described i the System Status Page o page 137. Carrier Gas is described below. HP6890 Colum Page Figure 5.19 HP6890 Colum page Legth Eter the colum legth i meters. Iteral Diameter Eter the iteral diameter of the colum i micrometers. Film Thickess Eter the thickess of the colum coatig i micrometers. Carrier Gas Select a carrier gas from the drop dow list box. Ijectio Pressure Mode Select Costat Pressure, Ramped Pressure, Costat Flow or Ramped Flow from the drop dow list box. Ijectio Port Select Frot or Back from the drop dow list box. Page 141

146 HP6890 Pressure Ramps Page Figure 5.20 HP6890 Pressure Ramps page This appearace of this page will vary depedig o the Ijectio Pressure Mode selected o the Colum page. If Costat Pressure was selected oly Iitial Pressure ad Iitial Time are eabled. If Ramped Pressure was selected the the Ramps are eabled as well. If Costat Flow or Ramped Flow was selected, the parameters will be as for the correspodig Pressure page but Pressure will be replaced by Flow. Iitial Pressure/Flow Eter the pressure/flow required for the Iitial state. Iitial Time Eter the legth of time to remai at the iitial pressure/flow Ramps To eable a Ramp check the relevat Ramp box or eter a o-zero value i the Rate box. Rate Eter rate of pressure/flow chage for curret ramp. Temp Eter fial pressure/flow for curret ramp. Time Eter the legth of time to remai at the fial pressure/flow of curret ramp, before proceedig to the ext ramp. Page 142

147 HP6890 Ove Page Figure 5.21 HP6890 Ove page Max Ove Temp Eter the maximum ove temperature. Temperatures etered i the ramps o the other pages will ot be accepted if they exceed this parameter. Equilibrium Time Eter the time to wait at the Iitial Temperature (defied o subsequet pages) before the ready sigal is displayed. Cryo coolig parameters To eable these parameters check the Eabled box. Cryo Uses CO2 Check this box if the cryo system uses CO 2 istead of N 2. Blast Coolig Eabled Check this box if the cryo system is to be used above ambiet temperatures to speed up coolig of the ove. For a detailed descriptio of Cryo Fault & Timeout cosult your HP6890 maual or site egieer. Ambiet Temp Eter the temperature regarded as ormal ambiet aroud the GC. Page 143

148 HP6890 Ove Temp Ramps Page Figure 5.22 HP6890 Ove Temperature Ramps page Iitial Temperature Eter the temperature required for the Iitial State. Iitial Time Eter the legth of time to remai at the iitial temperature. Ramps To eable a Ramp check the relevat Ramp No box or eter a o-zero value i the Rate box. Rate Eter rate of temperature chage for curret ramp. Temp Eter fial temperature for curret ramp. Time Eter the legth of time to remai at the fial temperature of the curret ramp, before proceedig to the ext ramp. Page 144

149 HP6890 Post Ru Page Figure 5.23 HP6890 Post Ru page This appearace of this page will vary depedig o the Ijectio Pressure Mode selected o the Colum page. If Costat Pressure or Ramped Pressure was selected the a Post Ru Pressure is required. If Costat Flow or Ramped Flow was selected the a Post Ru Flow is required. Post Ru Eabled Check this box if a Post Ru is required. Time Eter the legth of time for the post ru phase. Temp Eter the temperature for the post ru phase. Pressure/Flow Eter the head pressure/flow for the post ru phase. Page 145

150 HP6890 Frot ad Back Ijector Pages Figure 5.24 HP6890 Frot Ijector page Ijectio Port Type If Noe is selected there are o parameters to eter. Cool o Colum If Track Ove is selected there are o parameters to eter. If Ramped Temperature is selected Figure 5.24 is displayed. Thermal Zoe O Check this box to eable the Temperature, Time ad Ramps fields. The Iitial/Static coditios ad Ramps details are used to provide a temperature profile for the ilet idepedet to the temperature profile of the ove, but the method of use is the same. Iitial Temperature Eter the temperature required for the Iitial State. Iitial Time Eter the legth of time to remai at the iitial temperature. Ramps To eable a Ramp check the relevat Ramp box or eter a o-zero value i the Rate box. Rate Eter rate of temperature chage for curret ramp. Fial Temp Eter fial temperature for curret ramp. Time Eter the legth of time to remai at the fial temperature of the curret ramp, before proceedig to the ext ramp. Page 146

151 Split/Splitless If Noe is selected there are o parameters to eter. If Split is selected Figure 5.25 is displayed. Figure 5.25 HP6890 Frot Ijector page - Split Gas Saver Eabled Check this box to save gas after a ijectio. Note: Auto prep ru must be maually set to ON if this box is checked. See the HP6890 maual for details. Flow Eter the reduced flow rate. Time Eter the legth of time to deliver the reduced flow rate for. Thermal Zoe O Check this box to eable the Temperature ad Split Ratio fields. Temperature Eter the temperature at which to hold the ilet durig the ru. Split Ratio Eter the split ratio for the ilet flow. Page 147

152 If Splitless is selected Figure 5.26 is displayed. Figure 5.26 HP6890 Frot Ijector page - Splitless Gas Saver Eabled Check this box to save gas after a ijectio. Note: Auto prep ru must be maually set to ON if this box is checked. See the HP6890 maual for details. Flow Eter the reduced flow rate. Time Eter the legth of time to deliver the reduced flow rate for. Thermal Zoe O Check this box to eable the Temperature, Purge Time ad Purge Flow fields. Temperature Eter the temperature at which to hold the ilet durig the ru. Purge Time Eter the time at which to ope the purge valve. Purge Flow Eter the flow rate to use at the Purge Time. Page 148

153 PTV If Noe is selected there are o parameters to eter. If ay other optio is selected Figure 5.27 is displayed. Figure 5.27 HP6890 Frot Ijector page PTV Cryo Parameters Eabled Check this box eable the other optios o this dialog. Quick Cool Check this box to Eter the reduced flow rate. Time Eter the legth of time to deliver the reduced flow rate for. Thermal Zoe O Check this box to eable the Temperature ad Split Ratio fields. Temperature Eter the temperature at which to hold the ilet durig the ru. Split Ratio Eter the split ratio for the ilet flow. Max Ove Temp Eter the maximum ove temperature. Equilibrium Time Eter the time required to reach equilibrium at the ew temperature. Cryo coolig parameters To eable these parameters check the Eabled box. Cryo Uses CO2 Check this box if the cryo system uses CO 2 istead of N 2. Blast Coolig Eabled Check this box if the cryo system is to be used above ambiet temperatures to speed up coolig of the ove. For a detailed descriptio of Cryo Fault & Timeout cosult your HP6890 maual or site egieer. Page 149

154 Ambiet Temp Eter the temperature regarded as ormal ambiet aroud the GC. Flow Eter the reduced flow rate. Time Eter the legth of time to deliver the reduced flow rate for. Thermal Zoe O Check this box to eable the Temperature, Purge Time ad Purge Flow fields. Temperature Eter the temperature at which to hold the ilet durig the ru. Purge Time Eter the time at which to ope the purge valve. Purge Flow Eter the flow rate to use at the Purge Time. The Back Ijector parameters are edited i the same way as those for the Frot ijector. Page 150

155 PTV Settigs Page Figure 5.28 HP6890 PTV Settigs page Thermal Zoe O Check this box to eable the Temperature, Time, Ramps ad Pulsed Splitless Operatio fields. The Iitial/Static coditios ad Ramps details are used to provide a temperature profile for the ilet idepedet to the temperature profile of the ove, but the method of use is the same. Iitial Temperature Eter the temperature required for the Iitial State. Iitial Time Eter the legth of time to remai at the iitial temperature. Ramps To eable a Ramp check the relevat Ramp box or eter a o-zero value i the Rate box. Rate Eter rate of temperature chage for curret ramp. Fial Temp Eter fial temperature for curret ramp. Time Eter the legth of time to remai at the fial temperature of the curret ramp, before proceedig to the ext ramp. Gas Saver Eabled Check this box to save gas after a ijectio. Note: Auto prep ru must be maually set to ON if this box is checked. See the HP6890 maual for details. Flow Eter the reduced flow rate. Time Eter the legth of time to deliver the reduced flow rate for. Pulsed Pressure Eter the temperature at which to hold the ilet durig the ru. Pulsed Time Eter the time at which to ope the purge valve. Page 151

156 Purge Time Eter the time at which to ope the purge valve. Purge Flow Eter the flow rate to use at the Purge Time. HP6890 Commuicatio Parameters Select View Comms Settigs from the HP6890 Meu or press the butto to view the curret commuicatios settigs. toolbar Commuicatio settigs should oly be chaged by a egieer, the Edit Comms Settigs ad the toolbar butto allow this to be doe. Figure 5.29 HP6890 Commuicatios Parameters dialog Page 152

157 The HP6890 Toolbar The HP6890 toolbar has five extra buttos o it, which are: Click To View curret commuicatios settigs. Edit curret commuicatios settigs. Start ad stop method. Tur GC o ad off. Reset Autosampler Page 153

158 Page 154

159 Notes Page 155

160 Notes Page 156

161 Jasco Systems Jasco 900 ad Jasco 1500 Autosamplers Chapter 6 The Ilet Editor for the Jasco 900 ad Jasco 1500 autosamplers is the same. The Jasco 900 is used i the followig examples. Jasco Sampler Iitial Coditios Page This page is used to set parameters specific to the Sampler, to access it select Jasco900 AutoSampler from the View meu or press the toolbar butto. Figure 6.1 Sampler Iitial Coditios page Ijectio Volume Eter the volume i microlitres to iject. Note: If a multisample acquisitio is beig ru from the MassLyx Sample List, the ijectio volume defied i the sample list overrides the value defied here. Flushes Eter the umber of times the eedle should be flushed betwee ijectios. Aalysis Time Eter the legth of time the ru will last for. Vial umber Eter the umber of the vial to iject from. Note: If a multisample acquisitio is beig ru from the MassLyx Sample List, the Bottle # etry i the sample list overrides the value give i the Vial Number etry above. Page 157

162 Jasco 900 ad Jasco 1500 Pumps The Ilet Editor for the Jasco 900 ad Jasco 1500 pumps is the same. The Jasco 900 is used i the followig examples. The Jasco Pump pages ca be accessed by selectig Jasco900 Pump from the View meu o the Ilet Editor or by pressig the toolbar butto. Jasco Iitial Coditios Page Figure 6.2 Iitial Coditios page Solvets Up to three solvets will be displayed depedig upo the system cofiguratio. The total value of all the solvet percetages added together must ot exceed 100%. Pump A This is the remaider percetage after the solvet percetages have bee set for the other pumps. Pump B ad C Check the box for the pump required ad eter the percetage of flow to deliver from this pump. To disable the pump, ucheck the box. Flow This is the total flow rate of the solvet chaels. Mode Select Isocratic (Oe pump), Biary (Two pumps), HPG (High Pressure Gradiet) or LPG (Low Pressure Gradiet). Pressures Eter the upper ad lower limits of the pressure withi the solvet delivery system (SDS), if the pressure falls outside of this rage the SDS switches off. Colum Heater If the istrumet has a ove preset the the colum temperature ca be set to a specified temperature i degrees cetigrade. Check the Colum Temperature box ad eter a temperature. If the software has bee cofigured to operate without a colum ove the these boxes will be greyed out. Page 158

163 Stop Time Eter the time i miutes that the method will ru from the poit of ijectio. If a Jasco Autosampler has also bee selected, Aalysis Time o the autosampler page overrides this value. Post Time Eter the time i miutes that the istrumet will ru i its iitial coditios after a method has completed. No further ijectios ca be carried out whilst the system is i postru thus allowig re-equilibratio of the colum. Jasco Gradiet Page Figure 6.3 Gradiet Timetable page This page allows a gradiet to be etered ad edited. If isocratic mode was selected o the Iitial Coditios page the all fields will be greyed out ad is displayed. To add a gradiet, eter a time ad percetage i the relevat boxes ad press the toolbar butto. Note: The first etry must have a time of 0. To delete a sigle gradiet, click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To modify a gradiet select the required etry i the timetable. The values will the be displayed i the edit boxes to the right of the timetable ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. Note: If the time is chaged ad the ew time does ot correspod with a existig etry i the table, the a ew etry will be added. If the ew time correspod to a existig etry the the etry at that time will be overwritte. Page 159

164 Jasco 900 ad Jasco 1500 UV Detectors The Ilet Editor for the Jasco 900 ad Jasco 1500 UV detectors is the same. The Jasco 900 is used i the followig examples. This page is used to set parameters specific to the UV detector, to access it select Jasco900 UV Detector from the View meu or press the toolbar butto. Figure 6.4 UV Detector Cofiguratio page Respose This ca be set to Fast, Stadard or Slow depedig o the legth of time you expect the peak to appear. Wavelegth Set to the wavelegth you wat to moitor. Page 160

165 Notes Page 161

166 Notes Page 162

167 Shimadzu Systems Shimadzu Systems Chapter 7 Shimadzu Autosampler Iitial Coditios Page This page is used to set parameters specific to the Sampler, to access it select Shimadzu AutoSampler from the View meu or press the toolbar butto. Figure 7.1 Autosampler Iitial Coditios page Ijectio Volume Eter the volume i microlitres to iject. Note: If you are ruig from the Sample List, the ijectio volume i the sample list etry overrides the settig used here. Vial umber The vial to iject from. Note: If a multisample acquisitio is beig ru from the MassLyx Sample List, the Bottle # etry i the sample list overrides the value give i the Vial Number etry above. Rack Select Select the type of rack required from the drop dow list box. Syrige Volume Select the size of the curretly istalled syrige from the drop dow list box. Needle Stroke Adjusts the depth of the eedle tip to accommodate for sedimeted samples or o-stadard vials. Rise Volume Eter the volume of solvet that is to be rised through the eedle. Page 163

168 Risig Speed Eter the speed at which the solvet is to be rised through the eedle. Samplig Speed Eter the rate i microlitres per secod at which sample is extracted ito the autosampler eedle. This should be set accordig to the viscosity of your sample. Excess Volume To esure that the sample is ot diluted with the rise solvet more sample is draw ito the eedle tha will be ijected. Eter the extra volume required. Cooler Temperature If the sample cooler is istalled, eter the temperature that the sample should to be cooled to. MPT Tray Select Dual or Sigle from the drop dow list box. To Set-up Commuicatio Parameters (Shimadzu) Select Cofiguratio from the Shimadzu meu to display the Cofiguratio dialog. Figure 7.2 Cofiguratio dialog These parameters should be defied o setup ad should oly eed chagig if the Pump, Autosampler or mode of acquisitio is chaged. To chage a value select a ew oe from the relevat drop dow list box. Page 164

169 Shimadzu Pump The Shimadzu Pump pages ca be accessed by selectig Shimadzu Pump from the View meu o the Ilet Editor or by pressig the toolbar butto. Shimadzu Iitial Coditios Page Figure 7.3 Iitial Coditios page Solvets Up to four solvets will be displayed depedig upo the system cofiguratio. The total value of all the solvet percetages added together must ot exceed 100%. Pump A This is the remaider percetage after the solvet percetages have bee set for the other pumps. Pump B, C ad D These ca either be eabled or disabled by checkig the box ext to the idividual pumps. The values ca be set to the required percetage of flow delivery. Flow Rate This is the total flow rate of the solvet chaels accordig to how you have cofigured the istrumet. Eter High Pressure Limit ad Low Pressure Limit values as required. If the pressure falls outside these limits the curret acquisitio will stop ad the LC Status error light, o the MassLyx scree, will tur red. Colum Temperature Set Eter the temperature to heat the colum to. Note: This box will be greyed out if a colum heater is ot preset. Colum Temperature High Limit This is the maximum deviatio i colum temperature allowed. If this is exceeded the curret acquisitio will stop ad the LC Status error light, o the MassLyx scree, will tur red. Ru Time Eter the time i miutes that the method will ru from the poit of ijectio. Page 165

170 Shimadzu Gradiet Timetable Page Figure 7.4 Gradiet Timetable page This page allows a gradiet to be etered ad edited. To operate i isocratic mode esure that the timetable is empty. To add a gradiet, eter a time ad percetage i the relevat boxes ad press the toolbar butto. Note: The first etry must have a time of 0. To delete a sigle gradiet click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To modify a gradiet select the required etry i the timetable. The values will the be displayed i the edit boxes above the timetable, ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig will result i a ew etry beig created i the timetable. Flow Eter the flow rate for the solvet delivery system. Curve This sets the rate at which the solvet is to chage to the ew proportios ad/or flow rates. See the Shimadzu Operator s Guide for a list of values. Page 166

171 Shimadzu Iitial Evets Page Figure 7.5 Iitial Evets page This page allows the iitial state of switches 1 to 4 to be defied. Check the box(es) for the switches that should have a iitial state of off. Page 167

172 Shimadzu Programmed Evets Page Figure 7.6 Programmed Evets page This page allows the state of switches 1 to 4 to be programmed. To add a evet, eter a time, select a evet from the drop dow list box, select a actio (o or off) ad press the toolbar butto. To delete a sigle evet, click with left mouse butto o a time i the list ad press the toolbar butto. To delete all etries press the toolbar butto. This butto is oly available whe there are etries i the timetable. To modify a evet, select the required etry i the timetable. The values will the be displayed i the edit boxes above the timetable, ad ca be altered as appropriate. Oce chaged press to re-eter the values ito the timetable. If, however, you modify the time value such that it does ot correspod to ay existig etry i the timetable pressig will result i a ew etry beig created i the timetable. Page 168

173 Notes Page 169

174 Notes Page 170

175 CTC, Cetac ad Other Systems Chapter 8 CTC A200S Autosampler These pages are used to set parameters specific to the Sampler, to access them select CTCA200S AutoSampler from the View meu or press the toolbar butto. CTC A200S Status Page Figure 8.1 CTC A200S Status page The Fault, Ruig ad Ready idicators at the left side of the scree give iformatio o the curret status of the autosampler. Idicator Red Gree Fault Fault with the autosampler No fault Ruig Not ruig Ruig Ready Not ready Ready Page 171

176 Vial Eter the umber of the vial to take the sample from, for a sigle ijectio. Note: Whe samples are acquired from the Sample List the umber o the Sample List overrides this value. Press the butto to dowload the parameters to the LC system. Pressig the butto or choosig Load Method from the LC or CTC200 meu will perform the same actio. Press the butto to ru a sigle ijectio. Pressig the butto or choosig Ru Method from the LC or CTC200 meu will perform the same actio. CTC A200S Parameters Page Figure 8.2 CTC A200S Parameters page Sample Volume Eter the volume of sample (i microliters) to iject. Air Volume Eter the volume of air (i microliters) to be draw ito the eedle before the sample, to separate it from the previous sample. Pull-up Volume Eter the volume of sample (i microliters) to draw ito the eedle for a sample wash. Sample Washes Eter the umber of times to wash the eedle with sample. Solvet Washes Pre Ijectio Eter the umber of solvet washes to perform usig solvet from reservoirs A ad/or B, before a ijectio. Solvet Washes Post Ijectio Eter the umber of solvet washes to perform usig solvet from reservoirs A ad/or B, after a ijectio. Pull-up Cout Eter the umber of times to pull up the Pull-up volume for a sample wash. Page 172

177 Pull-up Speed Eter the speed (i microliters per secod) to pull up the Pull-up volume for a sample wash. Pull-up Delay Eter the time to wait betwee each pull up. Ijectio Speed Eter the speed (i microliters per secod) to iject the sample. Pre Ijectio Delay Eter the time to wait (i secods) betwee the eedle beig ijected ad the pluger beig depressed. Pre Ijectio Delay Eter the time to wait (i secods) for the pluger to be draw back after a ijectio. Page 173

178 CTC PAL Autosampler These pages are used to set parameters specific to the Sampler, to access them select PAL_CC AutoSampler from the View meu or press the toolbar butto. Whe the autosampler parameters are selected the followig message is displayed. Figure 8.3 Error message This refers to the Macro Editor, which is ot part of the stadard Cycle Composer software, see the PAL Cycle Composer User Maual for details. Press the Cacel butto to proceed. Whe the software is istalled a series of files are copied to the default.pro/acqudb directory (*.pma ad *.pol). Whe the autosampler page is selected the software looks for the presece of the PAL autosampler. If oe is foud iformatio is read from the Latest_pal.pol file. To create method editor files whe ot coected to a PAL copy the Offlie_pal.pol file to the Acqudb directory of the required project. Whe complete, copy the Method files (*.ccp) to the Acqudb directory of the required project o the acquisitio PC. Page 174

179 PAL Cycle Composer Method Editor Figure 8.4 Pal Cycle Composer Method Editor dialog To Create a Method (CTC PAL) 1. Select New Method from the File meu. The Choose ame of ew Method dialog is displayed. Figure 8.5 Choose ame of ew Method dialog 2. Eter the ame for the method ad press OK. 3. Eter a descriptio of the method i the Method Descriptio box. 4. Select the size of the syrige istalled from the Method Syrige drop dow list box. Note: The syrige size should be defied before ay macros are selected as differet default values ad rages are defied for each syrige size. Chagig the syrige size after selectig macros could result i the values etered beig outside the rages allowed ad so the macro values will have to be adjusted. 5. Select a macro from the Selected Macro drop dow list box. The parameters required for the macro are displayed below the selected macro box. Note: Placig the cursor o the macro ame will display a short descriptio of the macro fuctio, as i Figure 8.6. Page 175

180 Figure 8.6 Macro descriptio Placig the cursor o a field will display a descriptio of the valid values for this field, as i Figure 8.7. Figure 8.7 Macro descriptio 6. Whe the parameters for the macro have bee etered press the Isert butto ad the macro will be added to ed of the list i the Method Macro Sequece box. To replace a macro, click with the left mouse butto o the macro i the Method Macro Sequece box, select the macro to replace it with, defie the required values ad press the Replace butto. To remove a macro from the list, click with the left mouse butto o the macro i the Method Macro Sequece box ad press the Delete butto. 7. Select Save Method or Save Method As from the File meu. The Save Method dialog is displayed with the ame defaulted to that etered i step 2. The ame ca be chaged if required, ay chages made will be reflected i the Method Name i the editor. Press the OK butto to save the method. Figure 8.8 Save Method dialog 8. The method file (*.ccp) is stored i the Acqudb directory of the curret project. Page 176

181 To Modify a Method (CTC PAL) 1. Select the method required from the Method Name drop dow list box. The method files (*.ccp) displayed are those stored i the Acqudb directory of the curret project. 2. Click with the left mouse butto o the required macro i the Method Macro Sequece list ad chage the parameters displayed o the right of the dialog. Note: If the syrige size is chaged macro values will have to be adjusted as differet syrige sizes have differet field values. 3. To add a macro, select the macro from the Selected Macro drop dow list box. The parameters required for the macro are displayed below the selected macro box. Whe the parameters for the macro have bee etered press the Isert butto ad the macro will be added to ed of the list i the Method Macro Sequece box. To replace a macro, click o the macro i the Method Macro Sequece box, select the macro to replace it with, defie the required values ad press the Replace butto. To remove a macro from the list, click o the macro i the Method Macro Sequece box ad press the Delete butto. 4. Select Save Method or Save Method As from the File meu. The Save Method dialog is displayed with the ame defaulted to that etered i step 2. The ame ca be chaged if required, ay chages made will be reflected i the Method Name i the editor. Press the OK butto to save the method. Figure 8.9 Save Method dialog 5. The method file (*.ccp) is stored i the Acqudb directory of the curret project. To Delete a Method (CTC PAL) 1. Select the method required from the Method Name drop dow list box. 2. Select Delete Method from the File meu ad press Yes o the cofirmatio dialog. Note: Methods ca also be deleted usig Widow Explorer. Method files (*.ccp) are stored i the Acqudb directory of the curret project. Page 177

182 Sample List Vial Referecig (CTC PAL) For the CTC PAL autosampler the default Sample List vial referecig for a 96 well plate i Stack 1 is Stk1-01:1 to 96 for the first tray i stack 1 Stk1-02:1 to 96 for the secod tray i stack 1 etc. I.e. Stk1-01:1 is etered i the SAMPLE_LOCATION (Bottle) colum of the Sample List. To use the ormal MassLyx referecig 1:1, 1:2, 2:1 etc. the trays have to be reamed. See page 179 for details. Note: For OpeLyx the 1:1 tray:vial referecig must be used. Page 178

183 Usig the PAL CTC Autosampler with OpeLyx Whe usig the CTC PAL autosampler with OpeLyx the tray ames must be defied umerically ad i sequece. Note: It is recommeded that MassLyx is closed dow whilst chages are made to the tray umberig. Whe MassLyx is restarted the chages will be picked up automatically. To Reame Trays (CTC PAL) Usig the had held cotroller, check the order of the tray holders. Usig the had held cotroller, umber the trays for the tray holders sequetially startig from 1. E.g. if the tray holders are i the order: Stack1 THldr1 The the trays should be umbered startig at 1 for the first tray i stack 1 ad cotiuig sequetially for THdr1 trays (see the example i Table 8.1 ). Tray Holder Tray Type Default Tray Name OpeLyx Tray Name Stack1 MT96 Stk-01 1 Stack1 MT96 Stk-02 2 Stack1 MT96 Stk-03 3 Stack1 MT96 Stk-04 4 Stack1 MT96 Stk-05 5 Stack1 MT96 Stk-06 6 Stack1 MT96 Stk-07 7 Stack1 MT96 Stk-08 8 Stack1 MT96 Stk-09 9 Stack1 MT96 Stk Stack1 MT96 Stk Stack1 MT96 Stk THldr1 VT98 THldr THldr1 VT78 THdlr Table 8.1 OpeLyx Tray Namig Page 179

184 To check that trays are umbered correctly select OpeLyx Plate Logi from the Ilet Editor, Plate Logi meu. If the trays are correctly umbered the the dialog will appear as i Figure If the umberig is icorrect i.e. trays for THdlr1 are umbered from 1 the the dialog will appear as i Figure Figure 8.10 OpeLyx Plate Logi dialog Figure 8.11 OpeLyx Plate Logi dialog Usig Plates for OpeLyx Plate Logi (CTC PAL) Select OpeLyx Plate Logi from the Ilet Editor, Plate Logi meu. If a plate is labelled Disabled it ca be used for sigle shot logi. If a plate is labelled Eabled it ca be used for plate logi. To chage the state double click o the tray umber. Page 180

185 Cetac ASX100 Autosampler Figure 8.12 Cetac ASX100 Setup dialog Techical details of this autosampler are to be foud i the maual supplied with the Cetac ASX 100 autosampler. Vial Number Eter the umber of the vial to take the sample from. Note: Whe samples are acquired from the Sample List the umber o the Sample List overrides this value. Stadard Tube Check this box if stadard vials are beig used. The rack codes sectio will be grayed out ad vial umber ca oly be 2 to 14. Uptake Time Eter the time i secods for the sample to travel from the sample vial to the Mass Spectrometer. Before Sample Check this box to rise the eedle before each sample. Rise Time Eter the time i secods required to rise the eedle. Rack Codes Click with the left mouse butto o the code required for the rack. Page 181

186 Cetac ASX500 Autosampler Figure 8.13 Cetac ASX500 Setup dialog Techical details of this autosampler are to be foud i the maual supplied with the Cetac ASX 500 autosampler. Vial Number Eter the umber of the vial to take the sample from. Note: Whe samples are acquired from the Sample List the umber o the Sample List overrides this value. Stadard Tube Check this box if stadard tubig is beig used. The rack codes sectio will be grayed out ad vial umber ca oly be 1 to 10. Sipper Depth Eter the depth i millimeters the eedle should travel to. Note: The default probe samplig depth is measured from the eck of the tube to the tip of the probe. Uptake Time Eter the time i secods for the sample to travel from the tubig to the Mass Spectrometer. Rack Codes Click with the left mouse butto o the code required for each rack. Rise Before Sample Check this box to rise the eedle before each sample. Rise Time Eter the time i secods required to rise the eedle. Page 182

187 Solids Probe The temperature of a solids probe ca be cotrolled durig a acquisitio. To do this you must set a ramp that defies the temperature of the probe tip agaist retetio time. The ramp ca have up to 5 segmets which each have a start temperature, a time for which the probe will be held at that temperature, ad a rate at which the probe will be heated to reach the start temperature of the ext ramp segmet (if there is oe). I additio to these cotrols, TIC (Total Io Curret) cotrol of the probe is also available. If TIC cotrol is selected the the TIC value is moitored durig the acquisitio ad that iformatio is used to modify the programmed ramp i such a way that the system attempts to keep the TIC at or below the Maximum TIC value. This feature is very useful as it stops samples from beig burt off the probe prematurely. The actual temperature ramp used ca be stored with the data file ad a remote cotact closure ca be used to start the ramp. The Exteral Cotact Start box should be checked whe a robotic probe system is beig used. To Chage probe cotrol parameters (Solids Probe) 1. Choose Ilet from the Acquisitio Cotrol Pael Methods meu -or- Double click o the probe ico o the Acquisitio Cotrol Pael to brig up the solids probe editor show below. 2. Make ay chages to the parameters. 3. Save the method usig either Save or Save As from the File meu. Figure 8.14 Solids Probe Cotrol Editor Page 183

188 Programmig the temperature ramp (Solids Probe) The probe temperature ramp ca be programmed usig the keyboard or by draggig the small red hadles o the picture of the ramp itself. Usig TIC Cotrol (Solids Probe) DCI Probe To use the TIC cotrol feature you must first check the TIC Cotrol box, which will eable the values i the TIC cotrol group to be modified. There are 3 values that you the eed to set. Miimum sets the value for the TIC above which the probe rampig is reduced. If the actual TIC see from the istrumet is below this value, the full heatig rate, as programmed ito the temperature ramp is used to heat the probe. As the TIC rises above this value, the heatig rate is liearly adjusted dow based o the differece betwee this miimum value ad the maximum value discussed ext. For example, a TIC value exactly betwee the maximum ad miimum values would give a 50% rate compared to that requested by the ramp parameters. Maximum sets the value for the TIC at which the probe rampig is suspeded. This is doe because the system is tryig to keep the TIC at this level ad further heatig would cause it to rise above it. If the TIC reaches this level, heatig will ot recommece util it falls back dow below. Fiish sets a value for the TIC below which the acquisitio of data will termiate. The temperature program will cotiue however to allow ay remaiig sample to be burt off. A DCI probe ca be cotrolled i the same way as a solids probe, as described above with the exceptio that the ramp is programmed for Curret rather tha temperature ad the DCI curret ca be stored with the data file, ot the probe temperature. Thermospray Probe RoboProbe A thermospray probe ca be cotrolled i the same way as a solids probe, as described above, with the exceptio that the thermospray ozzle temperature ca be stored istead of the probe temperature. A robotic probe system ca be used i cojuctio with a CE Istrumets A200S auto ijector, or a Zymark laboratory robot. CE Istrumets A200S If the A200S is used the settig it up is doe as described earlier. Page 184

189 Zymark Labmate The Zymark setup editor allows you to select a solvet for your sample. 1. Choose Ilet from the Acquisitio Cotrol Pael Methods meu or Double click o the Picture of the auto ijector o the Acquisitio Cotrol Pael to brig up the Zymark editor show below. 2. Make ay chages to the parameters. 3. Save the method usig either Save or Save As... from the File meu. Cotact Closure Figure 8.15 Zymark Solvet Selectio Editor Cotact closure is a commo method of providig start/stop cotrol of a exteral ilet system. May chromatographs, both LC ad GC support cotact closure because it is ofte used to provide cotrol of a itegrator uit. MassLyx uses essetially the same method of sychroisatio for acquirig data, the mass spectrometer s cotrol uit usig the start ad stop sigals produced by the chromatography system to start ad stop data acquisitio. Page 185