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LC-NMR Accessory Installation

1. NMR Console Magnet LC hardware P table z a Sun host computer Y 0 25 inch 1 foot Room dimensions are about 6 1 m x 6 1 m 20 ft x 20 ft The standard NMR cabinet is 111 cm x 78 cm 44 in x 31 in Typical table size is 183 cm x 76 cm 72 in x 30 in The magnet is 97 8 cm 38 5 in in diameter Refer to the Installation Planning Guide for component dimensions Drawn approximately to scale 01 999036 00 C0800 LC NMR Accessory Installation 15 Chapter 1 Introduction LC NMR Suggested Room Layout 1 iS e Pi cat tt x 4 A NMR Console c y N I SS a E S S i v Sun host computer LC STAR workstation T 0 25 inch 1 foot Room dimensions are about 6 7 m x 7 3 m 22 ft x 24 ft The standard NMR cabinet is 111 cm x 78 cm 44 in x 31 in Typical table size is 183 cm x 76 cm 72 in x 30 in The magnet is 97 8 cm 38 5 in in diameter
2. 4 Make sure the wiring on the J2 connector is configured J2 Rear correctly wired as shown in the illustration on the right ro agi Black The J2 connector is incorrectly wired if the stop flow valve constantly rotates or the 16 port valve randomly increments Make sure the wiring matches the picture to the right Screen ii Red 01 999036 00 C0800 LC NMR Accessory Installation 21 Chapter 2 HPLC Hardware Setup Setting Up the ProStar 330 Detector Set up the ProStar 330 detector using the following procedures Unpacking the Detector and Removing Shipping Supports Installing the Analog Output Board Installing the High Pressure Flow Cell Loading the Control Software and Installing the ProStar 330 GPIB Card in the Dell Computer Loading Polyview Software Turning the Lamp On and Off Unpacking the Detector and Removing Shipping Supports 1 Remove the two transport screws as described by the instructions taped onto the cover of the instrument Figure 4 shows the location of the two transport screws on the bottom of the ProStar 330 detector Transport Screws 2 Position the ProStar E 330 on the bench Figure 4 ProStar 330 Transport Screw Locations and remove the side panel using the thumb screws 3 Remove the flow cell support from around the flow cell by reaching in and removing the supporting material by hand as shown in Figure 5 Figure 5 ProStar 330 Flow Cell Support Removal 4 Un
3. LC NMR Accessory Installation No 01 999036 00 Rev C0800 ave TAN VA AJ IAN LC NMR Accessory Installation Pub No 01 999036 00 Rev C0800 Revision history As 87 195280 00 A0496 Initial release B1096 added Analyte Collector and other changes B0397 added J2 voltage selector and interface box jumper settings C0997 updated for VNMR 6 1 added sample info for delay calibration C1297 added synch cable and 9065 detector As 01 999036 00 A0398 changed part number from 87 195280 00 to 01 999036 00 B0299 moved operation sections to User Guide Liquids and updated manual B0599 updates for Cascade delay time calibration and STAR software C0800 Add ProStar pump and detector update Cascade General updates Applicability of manual LC NMR Accessory for UN YINOVA UNITY plus and UNITY NMR spectrometer systems Technical contributors Chris Kellogg Karen Salomon Ron Haner Andy Myles Steve Smallcombe Paul Keifer Technical writer Dan Steele Mike Miller Technical editor Dan Steele Copyright 2000 by Varian Inc 3120 Hansen Way Palo Alto California 94304 http www varianinc com All rights reserved Printed in the United States The information in this document has been carefully checked and is believed to be entirely reliable However no responsibility is assumed for inaccuracies Statements in this document are not intended to create any warranty expr
4. RELAY1 J3 Stop flow valve RELAY 2 J4 RELAY 3 J5 REMOTE at SWITCHING ATTENTION Je 120 VAC OPERATION RELAYS 3 Synch Cable 03 919941 00 SWITCHING RELAY 6 wm READY OUT LE OUT mmm START IN mm FAULTIN SYNC SIGNALS mm FAULT OUT J 310 Figure 9 Diagram for Connecting the Stop Flow Cable with 310 Detector 28 LC NMR Accessory Installation 01 999036 00 C0800 2 5 Connecting the Stop Flow Cable e Figure 10 shows a diagram for connecting the stop flow cable to the 330 detector NMR console LC STAR workstation a interface box P4 P8 not used 330 LC Detector ProStar 330 Interface board 01 906664 0x FUSE TS 15AL250V Start out cable 03 935717 01 Do not connect P5 LC pump CI CI if using the analyte a N collector o i Power PE IRA e e E Stop flow valve ai cen ee 2 RELAY 2 J4 FA A rr 3ASB ATTENTION Lane WIRED FOR 120 VAC OPERATION REMOTE SWITCHING RELAYS MANUAL SWITCHING 2AMP 3A9 120V AC 50 60 HZ SYNC SIG
5. Collector loops L Aga do do dofa do Ade o gt e Agaelelaegelagegeae7 UU UUUUUGUGUUUUY Figure 29 Flow Diagram for On Flow 54 LC NMR Accessory Installation 01 999036 00 C0800 4 3 Analyte Collector Flow Diagrams NMR cell loading valve Syringe Yy Column Collector loops L Aga gd old do 4d dole Figure 30 Flow Diagram for Stop Flow Stopped NMR cell pump loading valve Injector valve waste amp Yy Switching Column Detector Collector loops 5 s a bsa bea kl be e Ke Figure 31 Flow Diagram for Loop Elution 01 999036 00 C0800 LC NMR Accessory Installation 55 Chapter 4 LC NMR Analyte Collector 4 4 Setting Up Software for Analyte Collector 1 Inthe Cascade program open up the system configuration window which is shown in Figure 32 2 Click on the analyte collector on line box as shown in 3 Click on Save as Default and then close the window EH System Configuration Figure 32 System Configuration Window 56 LC NMR Accessory Insta
6. Refer to the Installation Planning Guide for component dimensions Drawn approximately to scale 16 LC NMR Accessory Installation 01 999036 00 C0800 1 4 Site Layout Guidelines LC NMR Suggested Room Layout 2 lt s we LC hardware table o TE J NS w 7 Sun host computer LC STAR workstation iia Ty NMR Console l man fj yY ban 0 25 inch 1 foot Room dimensions are about 7 3 m x 7 3 m 24 ft x 24 ft The standard NMR cabinet is 111 cm x 78 cm 44 in x 31 in Typical table size is 183 cm x 76 cm 72 in x 30 in The magnet is 97 8 cm 38 5 in in diameter Refer to the Installation Planning Guide for component dimensions Drawn approximately to scale 01 999036 00 C0800 LC NMR Accessory Installation 17 Chapter 1 Introduction 1 5 Preparing Solvents This section provides information to consider when preparing solvents for LC NMR De gas or sparge solvents to prevent bubble formation in the probe cell Filter solvents that have been prepared by mixing and or dissolving one compound into another to remove residual or precipitated solid material Cover solvent conta
7. ASCII in STAR ProStar 330 Detector Method Window both the 9050 ProStar 310 go to the Print Options PEs Method Builder ProMethd mth l amp oen TUe aaia P age of Standard Cae poms DEV vie Pesii amp ProStar 310 UV Vis Detector Contro Reports Select the Sushoiruanees Convert results to i imo ASCII box Feinen lect th oe Unselect the Fort Automatic printing 5 romegar ome E Calibration Block Report Forr of reports amp peee z iew 2000 ciromatograms if ste na Posen you don t want a STAR printout of iin Prien A ibration Setup each run or if you Ee are not connected to the printer Figure 25 Detector Method Windows 6 To include a run log that has the pump and detector method parameters go to the Results format page under Standard Reports and select the Run log box 7 After you are finished entering parameters select Save or Save As from the File menu 8 Select Exit from the File menu 9 If necessary click on the System Control icon to return to the System Control window 10 Open the file menu and chose activate method A list of method files is displayed Choose the correct method and click on open 11 For the method to download to the pump the pump must be running Click on the pump button on the panel display 01 999036 00 C0800 LC NMR Accessory Installation 49 Chapter 3 Software Installation Cascade Drivers STAR 5 3 3 7 Setting Up the Ethernet Connection T
8. Cascade to a safe mode where all control buttons are inaccessible grayed out Subsequent installations of Cascade with the A D board drivers present might set this flag to false which means the flag must be set manually 3 6 Setting Up STAR 5 31 This section describes how to set up STAR 5 31 software e Loading STAR 5 31 this page e Building a Method in STAR 5 31 on page 48 e Building a Method in STAR 5 31 on page 48 Loading STAR 5 31 1 Install the STAR 5 31 software from the CD ROM a b ho ao h Insert the STAR 5 31 CD ROM and click INSTALL Select STAR Chromatography Work Station Read the license agreements and click Agree Enter name company and serial number from bright yellow sheet Install the software into the directory C STAR In the LC Components window select Example Data file and Advanced Applications If you have a 9065 detector click on 9065 driver box Click NEXT Answer No to the question Do you want STAR tool bar to start automatically Press RETURN and EXIT After the software is loaded a STAR icon appears in the left side of the screen 2 Load STAR Update 2 a b c f Insert the first of the three floppy disks Click on the FLOPPY A icon to open files Double click on the blue computer icon labeled set up If necessary enter A for the path The software begins to load At the prompt eject the disk and insert the second dis
9. Figure 16 Connect one end of the 1 8 inch PEM supply tube to the 1 8 inch hose adaptor 28 849916 00 Use a connector 28 859986 00 to connect the hose barb to the 1 4 inch tubing connected to the regulator Connect the other end of the 1 8 inch PEM supply tube to the IN connector on the back of the PEM You might need the connectors 28 8499 16 00 and 28 859986 00 Turn on the air flow 60 psi to the filter regulator assembly until the source air pressure is 60 psi Turn the knob on the regulator for the Powered Event Module until the pressure reads 60 psi The air line for the Powered Event Module is now installed 01 999036 00 C0800 LC NMR Accessory Installation 33 Chapter 2 HPLC Hardware Setup 2 7 Leak Testing the System This procedure describes how to perform a leak test of the after the stop flow valve and plumbing is installed A system leak test is available through the Extended Diagnostics of the 9012 LC pump Two leak test modes are provided constant pressure or pressure decay e In the constant pressure mode the pump operates to maintain a set pressure The reported leak rate in microliters per minute is the flow rate required to maintain the set pressure You should run the constant pressure mode leak test first e Inthe pressure decay mode the pump pressurizes the hydraulics to the set pressure and then stops The decay of the pressure is then monitored as a function of time The pressure decay mode is useful
10. Next Select Sound Video and Game Controllers for hardware type and click Next Dn wp Click the Have Disk button and browse to the A drive Go to the win95 directory 01 999036 00 C0800 LC NMR Accessory Installation 43 Chapter 3 Software Installation Cascade Drivers STAR 5 3 10 11 12 13 14 15 16 17 18 Select dt280 1 inf and click OK The window should now show A wind95 Click OK Select DT Open Layers DT2801 Series and click OK Click Finish Double click on the Multimedia icon in the Control Panel Select the Advanced tab Double click Media Control Devices Double click DT Open Layers DT2801 Series Device Driver Click the Settings button If you do not see DT2801 select ADD and add the dt2801 at hex address 0x2EC Under General Configuration change the DMA Channel number as follows For DELL GX1 Systems select DMA Channel 3 For other systems select DMA Channel 1 Click the Update button Click Close Driver Load Check Reboot and check that the drivers are loaded 1 2 Reboot the computer After the Win 98 picture is displayed the screen should go blank except for one line at the top of the screen that reads Voltdad 386 Data Transactions Data Acquisition This indicates that the board and drivers have been loaded properly This must be correct before loading Cascade 3 3 Installing Cascade The Cascade program coordinates the actions of the HPLC pump and stop flow
11. Out connectors A Injector Valve 5 2 6 PSE E k y NY 5 aN I EE COLLECTOR IN COLLECTOR OUT j A Transfer tube VTCONTAOL Red PEEK N out FROM ge et fo mse ee FROM PUMP TOWASTE a DETECTOR D LC NMR ANALYTE COLLECTOR 2 Waste LC Detector Figure 27 Analyte Collector Hydraulic Connections 01 999036 00 C0800 LC NMR Accessory Installation 53 Chapter 4 LC NMR Analyte Collector 4 3 Analyte Collector Flow Diagrams This section contains the following Analyte Collector flow diagrams e Collecting separated peaks Figure 28 e On flow Figure 29 Stop Flow stopped Figure 30 e Loop elution Figure 31 NMR cell Injector Pump loading valve valve waste lt v Switching valve Column Detector Y Collector loops Acie Q OKOLONO OOOO Figure 28 Flow Diagram for Collecting Separated Peaks NMR cell Pump loading valve Injector valve waste a Y Syringe Detector
12. PEM air requirements 31 Polyview 2000 software 24 Polyview Communications kit 25 Powered Event Module 31 preinstallation 13 preparing solvents 18 pressure decay mode 34 priming the pump 38 priming valve 38 probe drying prevention 18 probe flow cell flushing 39 ProStar 310 detector 21 ProStar 330 detector 22 prosthetic parts warning 7 Pulse Sequence Controller board 31 pumping solvent to probe 40 R radio frequency emission regulations 10 red PEEK tubing 19 regulator assembly 32 relay program 37 relays 37 relief valves warning 9 64 LC NMR Accessory Installation REMOTE SWITCHING 31 removable quench tubes warning 9 ribbon cable from ADC board 28 room layout considerations 14 S safety precautions 7 9 salts precipitated 18 setscout macro 61 setwet macro 6l site layout considerations 14 site requirements 13 solids high power amplifiers caution 10 solvent preparation 18 sound card conflict 45 source filter regulator air line 31 SPARE 1 30 STAR 5 3 software building a method 48 loading 46 STAR workstation 27 stop flow cable 28 stop flow valve 28 31 leak test 34 plumbing 25 sucrose requirements 13 T thumbwheel switch setting LC detector 31 LC pump 31 ProStar 310 detector 21 transfer tube 26 length 14 purging 40 requirements 18 transfering solvent to probe 40 transport screws 22 tubing cutter 25 V VEE samples loading 43 volumetric flasks requirements 13 VT experiment warning 8 W warnings d
13. diagrams for you to use as a guideline for setting up your LC NMR room The following room layouts are illustrated LC NMR Minimum Room Layout on page 15 LC NMR Suggested Room Layout 1 on page 16 LC NMR Suggested Room Layout 2 on page 17 Consider the following when developing a room layout 14 Stray magnetic fringe fields The Installation Planning Guide that came with your system contains magnetic field plots The distance between the NMR console and the magnet must be less than 3 m 10 ft to accommodate cable lengths between the console and the preamplifier housing magnet leg The transfer tube between the LC system and the probe is 4 6 m 15 ft no longer Therefore the distance between the LC hardware and the magnet should be about 3 m 10 ft to accommodate drops and rises The room must provide access for cryogen servicing of the magnet For cryogen servicing cryogen dewars need to be moved into the room near the magnet The LC STAR workstation PC should be set up close to the Sun host computers Cable lengths from the LC STAR workstation PC up to 3 35 m 11 ft between the LC workstation and the LC hardware up to 12 2 m 40 ft between the LC workstation and the NMR console Cable lengths from the Sun workstation and the NMR console up to 24 4 m 80 ft LC NMR Accessory Installation 01 999036 00 C0800 1 4 Site Layout Guidelines LC NMR Minimum Room Layout
14. eS Double click on the System icon in the Control Panel Click the Device Manager tab Select the Crystal PnP Audio MPU 401 Compatible entry Click the Properties button Click the Resources tab Remove the check by Use automatic settings Select Input Output Range Click the Change Settings button Use the scroll arrows to find an address that does not use 0330 033F e g 03E8 03E9 10 click OK three times to finish 3 5 Reinstalling Cascade for Windows 95 98 If it becomes necessary to reinstall Cascade use the steps in this section 1 Gw de gt o Restart the computer Select Add Remove Programs from Control Panel Select Cascade in the list of software Click on Add Remove Answer Yes to remove any shared files Restart the computer Ignore the warning The file vdtdad 386 cannot be found Select Add Remove Programs from Control Panel Choose Install and instert the first Cascade disk 01 999036 00 C0800 LC NMR Accessory Installation 45 Chapter 3 Software Installation Cascade Drivers STAR 5 3 Windows starts installing Cascade The message that Windows has detected a previous version of Cascade appears 8 Remove the disk when the installation is finished and restart the computer Note If Cascade is installed without drivers there is a flag in the registry editor that sets noboard to true The path is HKEY_CURRENTUSER Software VB and VBA Program Settings Cascade Hardware This limits
15. elucidation capabilities of NMR The HPLC and NMR systems are each complete and can be operated individually LC NMR integrates the two systems into a new class of analytical instrument Chromatography is an analytical technique that using the right conditions separates individual components from a mixture The LC NMR accessory includes the following components See Figure 1 e Microflow probe e Complete HPLC system including pump variable wavelength UV detector PC and column a peak sense module e LC NMR software added to VNMR which includes a variety of pulse sequences pulse shapes and macros for LC NMR data acquisition and data processing 1 1 About This Manual The installation instructions in this manual primarily cover the integration of the HPLC system with the NMR spectrometer The major components including the LC detector LC pump and valves of the HPLC system should be set up according to the Varian Chromatography Systems manuals The NMR spectrometer should be installed and running properly before being integrated with the HPLC system The first three chapters of this manual provide instructions for integrating the two systems 01 999036 00 C0800 LC NMR Accessory Installation 11 Chapter 1 Introduction 1 2 Compatibility The LC NMR accessory is fully compatible with YN TYZNOVA UNITYplus and UNITY NMR spectrometer systems with a Microflow probe WARNING Keep metal objects including tools electronic eq
16. floppy disks and tapes The field can also damage the strip of magnetic media found on credit cards automatic teller machine ATM cards and similar plastic cards Many wrist and pocket watches are also susceptible to damage from intense magnetism Refer to the manuals supplied with the magnet for the size of a typical 5 gauss stray field This gauss level should be checked after the magnet is installed 01 999036 00 C0800 LC NMR Accessory Installation 9 SAFETY PRECAUTIONS Caution Notices continued CAUTION CAUTION CAUTION CAUTION Keep the PCs including the LC STAR workstation beyond the 5 gauss perimeter of the magnet Avoid equipment damage or data loss by keeping PCs including the LC workstation PC well away from the magnet Generally keep the PC beyond the 5 gauss perimeter of the magnet Refer to the Installation Planning Guide for magnet field plots Check helium and nitrogen gas flowmeters daily Record the readings to establish the operating level The readings will vary somewhat because of changes in barometric pressure from weather fronts If the readings for either gas should change abruptly contact qualified maintenance personnel Failure to correct the cause of abnormal readings could result in extensive equipment damage Never operate solids high power amplifiers with liquids probes On systems with solids high power amplifiers never operate the amplifiers with a liquids probe The high power av
17. for further diagnostics if the system fails the constant pressure mode test Either leak test can be performed on the entire high pressure flow path not the probe When testing the entire system the plug must be placed upstream from the detector because most detector flow cells are not designed to operate at high pressure WARNING Hazardous chemicals are under pressure Use proper eye and skin protection CAUTION Do not leak test the probe or detector cell The high pressures used for the leak test will damage the flow cells in the probe and detector Never exceed 2 0 mL min flow rate through the probe Running the Leak Test The following steps describe how to run the leak test in either constant pressure or pressure decay mode 1 Carefully inspect the system for leaks drips or salt crystals Any obvious leaks must be corrected before continuing Locations to check include the piston seal drain the prime purge valve and all nuts and ferrules 2 Connect the solvent to be used for the leak test to the A proportioning valve Prime the pump and purge solvent through the entire system to remove any air These procedures are described later in Using the LC Pump on page 36 Choose a solvent compatible with the last solvent used in the HPLC system 3 Remove the fitting from the outlet side of the column Replace it with a known plug CAUTION The hydraulic path must be plugged in such a way that the pressure transducer is alwa
18. the same as the stop flow valve used to do The loop module is a detachable 16 port valve that can be moved between base units The 16 port valve has 130 uL 0 01 inch I D 1 16 inch O D PEEK tubing loops installed Larger or smaller loops can be installed 4 1 Connecting the Analyte Collector to the LC Workstation The Analyte Collector connects to the LC workstation with a terminal strip on the back of the base module This general purpose device uses contact closure to change valve positions Read back from the loop module occurs through BCD outputs To connect the Analyte Collector make the connections shown in Table 3 Disconnect the stop flow valve P5 when using the Analyte Collector 01 999036 00 C0800 LC NMR Accessory Installation 51 Chapter 4 LC NMR Analyte Collector Base eS g COLLECTOR IN COLLECTOR OUT FROM TO COLLECTOR FROM PUMP TO WASTE COLUMN aa EEJ e LC NMR ANALYTE COLLECTOR lt amp owm L towsste POWER FROM TO DETECTOR variano Figure 26 LC NMR Analyte Collector Table 3 Analyte Collector Wire Connections Pin Function Wire Color 1 To Column white green 2 To Column white orange or white only 3 To Collector waste none 4 To Collector waste white yellow 5 To Waste none 6 To Waste gray 7 To NMR jumpered from pin 18 8 T
19. tube to the 4 port on the stop flow valve Figure 8 3 5X Injector Valve 2 Detector Pump ES E 57399 6 DA COCCOLE o o CM see 2 8 ke Stop Flow oe eos eaqee Valve fg B080 5 A 5 3 sc Soest e ooo ES SS SS Transfer tube to probe Red PEEK Figure 8 Stop Flow Valve Plumbing Connection Diagram 3 For probe tests this end of the transfer tube is temporarily connected to the pump outlet port 26 LC NMR Accessory Installation 01 999036 00 C0800 The stop flow valve is now plumbed into the Varian HPLC system Go to the next section to set up the PC workstation The table on the right gives an alternative plumbing configuration that maintains pressure during stop flow 2 4 Unpacking and Assembling the LC STAR Workstation If the Microflow probe is installed connect the other end of the transfer tube to the LC connector on the probe labeled IN Otherwise place this end in a waste container near the magnet Connect tube from the 1 and 6 ports on the stop flow valve to the waste container Valve Port Connection plug from pump mixer to purge valve to NMR from detector Ne DN WwW to waste 2 4 Unpacking and Assembling the LC STAR Workstation Use the instructions provided in the Varian Chromatography Systems manuals to unpack and
20. under A or whatever inlet port is attached to the DO flushing solution 01 999036 00 C0800 LC NMR Accessory Installation 39 Chapter 2 HPLC Hardware Setup 4 Use the CURSOR buttons to move the cursor under Flow Use the INCREASE or DECREASE buttons to set the value under Flow to 1 00 This sets the pump rate at 1 0 mL min Press the RESET button twice Press PUMP A Pump the flushing solution through the probe for at least 3 minutes Co ND Examine the output of the probe for steady flow into the waste container 10 The flow characteristics will be either steady flow or drop wise depending on the solvent 11 After 3 minutes press STOP to stop the pump Purging the Transfer Tube This procedure describes purging the transfer tube of old solvent and filling it with new solvent The procedure is for a fully installed LC NMR system CAUTION To avoid unnecessarily high fluid pressures in the NMR probe never exceed 2 0 mL min flow rate through the probe The preferred flow rate is 1 0 mL min or less This requires extra attention during pump priming and purging procedures Beware of clogged or crimped output tubing Note Before starting make sure the probe is disconnected from transfer line 1 Select 100 A B or C for the specific inlet port desired Select a flow rate of 1 0 mL min and press the RESET key twice 2 Make sure the prime valve is open then press the PRIME key and prime the transfer line for a
21. 4 2 Installing the Analyte Collector Plumbing oo cece cee ceeceeeeeeeseeeeceeeeeeeeaeeees 53 4 3 Analyte Collector Flow Diagrams oo ee cceeeessessecseesseceecesecseceseeeeeeeeeseeeeeseeeaeeees 54 4 4 Setting Up Software for Analyte Collector sessessseeeieseiereeeersiseerssrsrerersrrrrererserenerees 56 Chapter 5 LC System Calibrations scccsssseceesseeeeeesseeeeeessneeeeenseees 57 5 1 Calibrating LCID WET gassiseivncriiiii dite ations diene k SEENE RESES 57 5 2 3C Sideband DUPPLCSSION ssesoccse dscevavens taaescecucsgesd oqavsea REEE AAEE RECE NOREA 58 5 3 Calibrating the Delay TIME cc ccccccscccassssssscessscssccssscnssaevonsssdscosdseiansessrseaiesbbosteaanstbesnie 58 Calibrating from the LC Detector to the Microflow Probe eessen 58 Calibrating from the LC Detector to the Analyte Collector wee eee 60 Calibrating from the Analyte Collector to the Microflow Probe eee 60 5 4 New Solvent Concentration cccccscccecssecessececenceceseececesaeceseeeecesaeceseaueceseeeeessneeeneaaes 61 lato Ce oc iatiscisueaasstsnsteusntshancsaasendnsnsagasniesnnsdesnantanssdsutuxedsakersnseesnsssancntuossteasensestdennans 63 4 LC NMR Accessory Installation 01 999036 00 C0800 List of Figures Fipure 1 LC NMR Block Diagram cs sc cc cesscsszetisskscnsstanesszacbaasespesesevessbancvonecasnscioaecnaessensscapespiasbadoe 12 Figure 2 Mounting the Injector and Purge Prime Valves eeeceeeecseseeeeecseeceeceesaecns
22. 9036 00 C0800 5 4 New Solvent Concentration 1 Load loops 1 to 10 with the methyl benzoate sample This requires 10 separate injections e Use an isocratic method 50 acetonitrile 50 DO 5 min end time 1 mL min 280 nm detection e Inject 25 mg on column 50 uL of 25 mg 50mL standard 2 Set the system up for analyte analysis a In System Control of STAR Chromatography activate the LC method used for analyte analysis in the previous calibration b In System Configuration of Cascade select Analyte Analysis c Set the hold time to 1 minute and make sure that the system is not waiting for the NMR to complete d Set the Threshhold to 200 mV e Set the End time to 2 minutes f Start with a from loop to probe delay set to 10 seconds This assumes that the LC method is at 1 mL mi Save the configuration g Make sure that the Inhibit apex detection is not selected on the main menu h Go to the analyze loops section in the analyte collector window set the wait time between runs to 3 to 4 minutes Start and end with loop 1 Close the analyte collector window i Click on Start Run in the main menu to start analyte analysis After the wait time is counted down loop will be emptied and a peak should be detected in the LC NMR stop flow program after it crosses the threshhold The delay time will be counted down and the system will stop while the NMR acquires data 3 After the hold time has expired the LC NMR run will con
23. NALS mm READY IN mm ENABLEIN mmm START OUT mm FAULT OUT Y cable 01 906684 0x lt Figure 10 Diagram for Connecting the Stop Flow Cable with 330 Detector 01 999036 00 C0800 LC NMR Accessory Installation 29 Chapter 2 HPLC Hardware Setup 1 Set the jumpers in the interface box see Figure 11 for either UN YINOVA UNITYplus or UNITY as follows a d Loosen the four screws on the side of the interface box Remove the interface box cover Set jumpers J2 and J3 for UNTYTNOVA UNITY plus or UNITY LT UNITYTNOVA or UNITYplus set J2 C and J3 to the side J3 nearest the U I Set to Set to label UNITY INOVA or UNITYplus UNITY set J2 and J3 to the side nearest Figure 11 Interface Box Jumper Settings the UNITY label Replace the cover and tighten the four screws ALINN oo hs I n I n ALINN 2 Connect the ribbon cable between the ADC board in the PC and the interface box 3 Connect the two cables to the NMR console as follows e UNITYTNOVA Connect one cable P3 to SPARE 1 on the Breakout panel Connect the other cable P4 to EXT TIME BASE on the DAC board Figure 12 shows the connectors on the Breakout panel N d seat J8215 u CLKSTB J8211 RCVR GATE S ACK e as ATTEN R R EMERG J8216 Jzs2 ack 19 S SPARE 1 A J232 STB J233 ACK 2 P J233
24. STB 3 E BEF 2 SE SPARE 2 O a 2 O J8218 P T eu E SPARE 3 S i R U S J8219 Sj T SPARE 4 A A 7 J8210 j i SPARE 5 J8233 J8201 J8213 Figure 12 VNITYZNOVA Breakout Panel Connectors 30 LC NMR Accessory Installation 01 999036 00 C0800 2 6 Installing the Powered Event Module e UNITYplus Connect one cable P3 to SPARE 1 on the Pulse Sequence Controller board Figure 13 Connect the other cable P4 to Ext Time Base on the Pulse Sequence Controller board 40 MHZ IN EXT TIME BASE NTHDLP i I puisescauence sO 8o 4 PF FULL LOT INGING Io SPARE 1 J3 CABLE 993019 CONTROLLER F OVERUN me TE lem FIFO EMPTY APTO Figure 13 UNITYplus Pulse Sequence Controller Board Front Panel UNITY Connect one cable P3 to SPARE 1 on the XL Interface board in the 12 pin jumper pack J5 connect one wire to pin 7 and one wire to ground Connect the other cable P4 to Ext Time Base on the Acquisition Controller board 4 Refer to Figure 9 for the 310 or Figure 10 for the 330 Then make the following connections a Connect the cable labeled P2 to J2 DATA SYSTEM OUTPUT on the detector b Connect the following cables according to the type of detector 310 detector connect the cable labeled P9 to J9 on the LC pump Connect the Sync cable 03 919941 00 between J9 on the 310 detector and J10 on the LC pump Connect the end labeled P9 9010 to J9
25. ailable from these amplifiers will destroy liquids probes Use the appropriate high power probe with the high power amplifier Take electrostatic discharge ESD precautions to avoid damage to sensitive electronic components Wear a grounded antistatic wristband or equivalent before touching any parts inside the doors and covers of the spectrometer system Also take ESD precautions when working near the exposed cable connectors on the back of the console Radio Frequency Emission Regulations The covers on the instrument form a barrier to radio frequency rf energy Removing any of the covers or modifying the instrument may lead to increased susceptibility to rf interference within the instrument and may increase the rf energy transmitted by the instrument in violation of regulations covering rf emissions It is the operator s responsibility to maintain the instrument in a condition that does not violate rf emission requirements 10 LC NMR Accessory Installation 01 999036 00 C0800 Chapter 1 Introduction Sections in this chapter e 1 1 About This Manual this page e 1 2 Compatibility page 12 e 1 3 Preinstallation Requirements page 13 e 1 4 Site Layout Guidelines page 14 e 1 5 Preparing Solvents page 18 e 1 6 Caring for the HPLC System and Microflow Probe page 18 The Varian LC NMR accessory combines the separation capabilities of high performance liquid chromatography HPLC with the structure
26. al stored in each loop 5 Doan analyze loops cycle a In STAR Chromatography create an isocratic method of 50 acetonitrile 50 D O with a 2 minute run time Set the detector wavelength to 280 nm b Inhibit injection by not setting the relays in the pump section of the method c Save the method and then go to System Control and Activate the method and wait for the ready lights on the pump and detector d Inthe System Configuration window of Cascade check Analyte Analysis e Set the Run end time to 2 minutes f Inthe Loop analysis section of the Analyte Collector window of Cascade set the start of analysis to loop 1 and finish with loop 7 Set a delay of 3 seconds and a wait of 4 minutes 6 Inthe Main menu data acquisition and display click on Start Run Select Inhibit apex detection 7 Each loop should be automatically emptied in turn and the UV spectrum recorded Record the apex intensity of the eluted compounds The correct delay time is the shortest time necessary for maximum intensity Typical values at 1 mL min are between 6 10 seconds Enter the value in the configuration window of the LC NMR stop flow program in the To collector loop window Calibrating from the Analyte Collector to the Microflow Probe Once the delay time from the UV detector to the analyte collector has been calibrated it is necessary to calibrate the delay time from the analyte collector to the flow probe LC NMR Accessory Installation 01 99
27. and possibly extensive damage to the magnet It could also cause a sudden dangerous release of nitrogen and helium gases from the dewar Except when transferring nitrogen or helium be certain that the relief valves are secured on the vent tubes WARNING On magnets with removable quench tubes keep the tubes in place except during helium servicing On Varian 200 and 300 MHz 54 mm magnets only the dewar includes removable helium vent tubes If the magnet dewar should quench sudden appearance of gases from the top of the dewar and the vent tubes are not in place the helium gas would be partially vented sideways possibly injuring the skin and eyes of personnel beside the magnet During helium servicing when the tubes must be removed carefully follow the instructions and safety precautions given in the manual supplied with the magnet Caution Notices Observe the following precautions during installation operation maintenance and repair of the instrument Failure to comply with these cautions or with specific cautions elsewhere in Varian manuals violates safety standards of design manufacturing and intended use of the instrument Varian assumes no liability for customer failure to comply with these precautions CAUTION Keep magnetic media ATM and credit cards and watches outside the 5 gauss perimeter from the centerline of the magnet The strong magnetic field surrounding a superconducting magnet can erase magnetic media such as
28. assemble the LC STAR workstation and to connect the GPIB IEEE interface cables to the pump and detector After the LC STAR workstation is assembled use the following procedures to install the ADC board in the PC connect the PC cables and install the LC Stop Flow Monitor software CAUTION Avoid equipment damage or data loss by keeping the LC STAR workstation PC well away from the magnet Generally keep the PC beyond the 5 gauss perimeter of the magnet Refer to the Installation Planning Guide for magnet field plots This procedure describes how to install the ADC board into the LC STAR workstation The ADC board in the PC provides the basic analog and digital I O interface between the LC STAR workstation LC hardware and the NMR system 1 Make sure the PC power is off but leave the power cord connected Follow anti static procedures because some of the parts in the PC and on the boards are static sensitive Remove the cover from the PC The PC contains a number of ISA type expansion slots One of the expansion slots should already contain the GPIB IEEE interface board The ADC board can be installed in any one of the other free expansion slots Choose the expansion slot for the ADC board If the expansion slot has a slot cover on the back panel of the PC remove it Locate the ADC board in the kit and carefully remove board from its packaging Be sure to hold the board only by its edges Install the ADC board in the empty sl
29. bout one minute 3 Press STOP Purging pumps the solvent at 10 mL minute at concentrations specified by the ABC FLOW proportions The purge operation ignores the specified flow rate and automatically pumps at 10 mL minute Transferring a Solvent into the Probe This procedure describes how to use the pump to transfer a solvent into the probe flow cell The procedure is for a fully installed LC NMR system 1 Ifthe transfer tube is not connected to the probe connect it to the IN port on the probe body 2 Press ABC FLOW The value 100 should already appear under whatever inlet port is attached to the solvent you want to transfer If not use the CURSOR buttons and the INCREASE or DECREASE button to set the value to 100 CAUTION To avoid unnecessarily high fluid pressures in the NMR probe never exceed 2 0 mL min flow rate through the probe The preferred flow rate is 1 0 mL min or less This requires extra attention during pump priming and purging procedures For the same reason the maximum 40 LC NMR Accessory Installation 01 999036 00 C0800 2 8 Using the LC Pump length of output tubing from the probe to the waste receptacle is one meter and do not use output tubing that is less than 0 010 inches inside diameter Beware of clogged or crimped output tubing 3 Use the CURSOR buttons to move the cursor under Flow 4 Use the INCREASE or DECREASE buttons to set the value under Flow to 1 00 This sets the pump rate at 1 0
30. cneeeseeneeenees 54 Figure 29 Flow Diagram for On FlOW eects cseeeseeseeeseesecesceseceseesescneeaecaeesaecsaesaecnessaeeneesnees 54 Figure 30 Flow Diagram for Stop Flow Stopped sssscsecssseeeeeceeceeeeeesecaecaeesesaeceeeeeeaeaeeees 55 Figure 31 Flow Diagram for Loop Elution cece ceecsesecseeeceeceseeeeeeceecseeseesecaesaeeseaecaseeeeaesaeeees 55 Figure 32 System Configuration Window sccscsssscsssecseeeeescsaecerseceecseeseesecaeceeescaecaeeeeeaesaseees 56 Figure 33 Profile Showing Delay Time is Correct ceeesscssscseeeseeceecseeseesecneceeesesaecaeeeesaesaeeees 59 Figure 34 Profile Showing the Delay Time is Too Short 0 eee eeeeeceseneeeeeeecneeseesesaecaeeecsaeeaeeees 59 Figure 35 Profile Showing the Delay Time is Too Long eee ceeeeceecneeeeeeeeeeceeeseeaeceeeeeeaeaeeees 59 01 999036 00 C0800 LC NMR Accessory Installation 5 List of Tables Table 1 Pump Method 1 Conditions Table 2 Initial Relay Program for Pump Method 1 o e eeceecceeseescecceeccesceeeeceeeesecacenaeceeeeeeneeeaees Table 3 Analyte Collector Wire Connections 6 LC NMR Accessory Installation 01 999036 00 C0800 SAFETY PRECAUTIONS SAFETY PRECAUTIONS The following warning and caution notices illustrate the style used in Varian manuals for safety precaution notices and explain when each type is used WARNING Warningsare used when failure to observe instructions or precautions could result in
31. d WARNING Do not look down the upper barrel Unless the probe is removed from the magnet never look down the upper barrel You could be injured by the sample tube as it ejects pneumatically from the probe WARNING Do not exceed the boiling or freezing point of a sample during variable temperature experiments A sample tube subjected to a change in temperature can build up excessive pressure which can break the sample tube glass and cause injury by flying glass and toxic materials To avoid this hazard establish the freezing and boiling point of a sample before doing a variable temperature experiment LC NMR Accessory Installation 01 999036 00 C0800 SAFETY PRECAUTIONS Warning Notices continued WARNING Support the magnet and prevent it from tipping over The magnet dewar has a high center of gravity and could tip over in an earthquake or after being struck by a large object injuring personnel and causing sudden dangerous release of nitrogen and helium gasses from the dewar Therefore the magnet must be supported by at least one of two methods with ropes suspended from the ceiling or with the antivibration legs bolted to the floor Refer to the Installation Planning Manual for details WARNING Do not remove the relief valves on the vent tubes The relief valves prevent air from entering the nitrogen and helium vent tubes Air that enters the magnet contains moisture that can freeze causing blockage of the vent tubes
32. e container under the drain line during the priming process 6 Press A to prime the inlet port A The pump starts the PRIME LED lights and the solution connected to inlet port A begins to flow through the prime valve into the waste container When the liquid flow from the prime drain is continuous and free of bubbles press STOP 7 Repeat step 4 for inlet ports B and C 8 After priming is finished close the prime valve by turning it clockwise 9 Reconnect the tubes that you disconnected in steps 1 and 2 a Disconnect the red tube from the bottom port on the LC pump and connect it to the 4 port on the stop flow valve b Reconnect the blue tube to the bottom of the LC pump LC NMR Accessory Installation 01 999036 00 C0800 2 8 Using the LC Pump Prime valve Drain LC pump output port amp Figure 21 Prime Valve and Drain on Front of LC Pump The pump is now ready to transfer solvents to the probe Flushing the Probe Flow Cell This procedure describes flushing out the probe flow cell To prevent contamination the probe flow cell must be flushed out with D30 after you finish using a particular solvent The procedure is for a fully installed LC NMR system 1 Connect the transfer line to the probe 2 Use the CURSOR buttons to move the cursor under A or whatever inlet port is attached to the bottle containing the D20 3 Press the INCREASE button until the value is 100
33. e relay program for method 1 is 3 100 finished Table 2 shows three lines of the relay program Priming the Pump This procedure describes how to prime the pump after new solvents are attached to the inlet ports The procedure is for a fully installed LC NMR system CAUTION To avoid unnecessarily high fluid pressures in the NMR probe never exceed 2 0 mL min flow rate through the probe The preferred flow rate is 1 0 mL min or less This requires extra attention during pump priming and purging procedures For the same reason the maximum length of output tubing from the probe to the waste receptacle is one meter and do not use output tubing that is less than 0 010 in inside diameter Beware of clogged or crimped output tubing 1 Disconnect the blue tube from the bottom port on the LC pump Figure 21 shows this port 2 Disconnect the red tube from the 4 port on the stop flow valve and connect it to the LC pump output port the bottom port on the LC pump shown in Figure 21 3 Disconnect the transfer tube from the probe and place the open end of the transfer tube into a waste container near the probe 4 Connect the new solvent bottles to the pump inlets using dry pieces of 1 8 inch tubing Inlet filters are not recommended for LC NMR Make sure each bottle is labeled and dated 5 Open the priming valve on the front panel of the pump see Figure 21 Turn the prime valve about one turn counterclockwise Place a wast
34. eaaeeeeeenaeeeeaeeeee Transferring a Solvent into the Probe oes eeeecesesceseeeeceececeseesecaecateeeeaeeneeees Chapter 3 Software Installation Cascade Drivers STAR 5 3 see Sl Loading the VEE Samples sc sscce iesessssive cssctasessdesceveseteseuceesnsevadsscceseusceccraueuedvsnsivercessees 3 2 Installing Software Drivers 2 i iniecldsh aiid esedde ea a ens aiT ei 01 999036 00 C0800 LC NMR Accessory Installation Table of Contents Driver Installation secccscissssavansserliieceisiseeuelinctboaces EEEE AE EE EREE 43 Driver Load Cheek o sces sascssustasebeasuevaxestapdocaghissjccessoneusenedeysnase dhvesssaseesisdgatagansees 44 Sedh Installame Cascade ssrneioie eana a E ts N RE E E 44 3 4 Resolving Sound Card Conflict on DELL Computers sssssesessesesesseseserereersreeresreseses 45 3 5 Reinstalling Cascade for Windows 95 98 oo ec eeeeeeeeseeseeeseeseecaecaeceeseseeeesseeeeees 45 30 Setting Up STAR 5 30 sareei ia AEE TEORA E E acces 46 Loading STAR 5 31 vi rr orrip e eraan aoa aaa EEB Naa sr AEE E Ea a ss dabatsa ee 46 Building a Method in STAR 5 31 eessssessessesisrsesreresrsrererererrersrsrrreesrsreresrrerens 48 3 7 Setting Up the Ethernet Connection ooo eee eeeecceeeeeeceseeeeeeseeteecaeecsecsuesaeenaeeaeens 50 Chapter 4 LC NMR Analyte Collector ansssnsnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn nnman 51 4 1 Connecting the Analyte Collector to the LC Workstation sesseeeseseeersesereerereerrerees 51
35. eeaeeneensees 20 Figure 3 310 Detector Voltage and Device Address Switches ecceeceseeseesecsseeecsseeseeneeesees 21 Figure 4 ProStar 330 Transport Screw Locations 0 ceeceesceseesceseeeeceseeeeecaeecaecaesaecsaeeaeenseeaees 22 Figure 5 ProStar 330 Flow Cell Support Removal 0 eeeeececeeceseeeeeseeeeecaeesaeceesaecneseseeneensees 22 Fipure 6 ProStar 330 Sponge Remiovall iis ccccssccsssvesescascecsansesscsoatessscesnebsvesevarescsnacnnsnesessensesessnneaee 23 Figure 7 ProStar 330 Analog Output Board Install eee ecceeeeeeeeeeeeecaeesaecaeeaeceenaeeneeeaees 23 Figure 8 Stop Flow Valve Plumbing Connection Diagram 000 cece eceeeeeecseeeeeseeeeaecneeeaeeneenaees 26 Figure 9 Diagram for Connecting the Stop Flow Cable with 310 Detector ieee eeeeeeteeeeees 28 Figure 10 Diagram for Connecting the Stop Flow Cable with 330 Detector cece eeeeeteeeeees 29 Figure 11 Interface Box Jumper Settings 20 0 0 eeeeseeeeceeeeseceseeececseeeeecaeesaecaaesaecnessaeeneeenees 30 Figure 12 UN YINOVA Breakout Panel Conmectors essseeeseeseeeseeceecseeeesecsecaesceaecaseeesesaeeees 30 Figure 13 UNITYplus Pulse Sequence Controller Board Front Panel eee eeeeeeeeeeeeeees 31 Figure 14 Powered Event Module Back Panel Connectors eee eceeeeecseessececeseecneeeseeneenaees 32 Figure 15 Injector Valve Top VIEW ssccccdesesesececcceesvsesecs secsssedscsareeccsscosusecideocs csesctcezouardecnsdvespisetvess 32 Figure 16 Connecting the Regulato
36. efined 7 waste container 13 Windows 95 44 X XL Interface board 31 Y yellow exclamation point 45 01 999036 00 C0800
37. essed or implied Specifications and performance characteristics of the software described in this manual may be changed at any time without notice Varian reserves the right to make changes in any products herein to improve reliability function or design Varian does not assume any liability arising out of the application or use of any product or circuit described herein neither does it convey any license under its patent rights nor the rights of others Inclusion in this document does not imply that any particular feature is standard on the instrument UNITYTNOVA MERCURY Gemini GEMINI 2000 UNITY plus UNITY VXR XL VNMR VnmrS VnmrX Vnmrl VnamrV VamrSGI MAGICAL II AutoLock AutoShim AutoPhase limNET ASM and SMS are registered trademarks or trademarks of Varian Inc Sun Solaris CDE Suninstall Ultra SPARC SPARCstation SunCD and NFS are registered trademarks or trademarks of Sun Microsystems Inc and SPARC International Oxford is a registered trademark of Oxford Instruments LTD Ethernet is a registered trademark of Xerox Corporation VXWORKS and VxWORKS POWERED are registered trademarks of WindRiver Inc Other product names in this document are registered trademarks or trademarks of their respective holders Table of Contents SAFETY PRECAUTIONS sisiiscacatscncsncsecessdesciamisesnsidieeduccaussieassniatessteasuedgancinasend Ghapter 1 Introd ction ssssiisssisisiniinninininnaninniius anakana saanunna enian 1 1 Abou
38. essure flow cell 1 4 Remove the low pressure flow cell that was shipped with the system Pull back the retaining ring and lift the flow cell out Gently remove the attached tubing from the supports and pull the unit away from the box Install the high pressure flow cell by pulling back the retaining ring and positioning the new flow cell in the light path Put the attached tubing into the side support Thread the inlet tubing out the inlet port and the outlet tubing through the outlet port on the front panel Attach the inlet and outlet tubing to the rest of the system with the PEEK fittings Do not overtighten because the tubing is made from Teflon and is easily crushed Replace the side cover Loading the Control Software and Installing the ProStar 330 GPIB Card in the Dell Computer The National Instruments control software must be loaded onto the Dell PC before the GPIB board is installed 1 gt SS ON 10 11 12 Insert the National Instruments CD into the CD ROM drive The standard Windows installation screen appears followed by a screen called Welcome to National Instruments GPIB Setup Choose Other and click on the Next button Accept the license and click Next on the line registrations screen Start the installation and select all of the components in the Select the Components window Click on Next Click Next on the Install screen After the installation is finished click Finish Remove
39. gth array 9 Enter gzlvlw gzlvlw n ga where n is the number of spectra with the best CH3CN suppression and minimal artifacts This number does not need to be recalibrated unless suppression or artifacts change significantly 10 Enter ssfilter y wft to turn on solvent subtraction of CH3 CN 01 999036 00 C0800 LC NMR Accessory Installation 57 Chapter 5 LC System Calibrations 5 2 13C Sideband Suppression This procedure describes how to decouple 13C sidebands during WET pulses 1 Tune the X channel of the probe to 5C connect the decoupler cable 2 Enter dmfwet 427 dpwrwet 21 3 Set dof to the approximate 13C shift of CH3CN 4 Set dmm or dseq for GARP1 decoupling CW mode is also acceptable 5 Set c13wet to one of the following cl3wet y eliminates 13C sidebands Data normally responds well to ssfilter 80 ssntaps 161 cl3wet n keeps 5C sidebands Data may look better with ssfilter n because of phase distortion of 13C sidebands 5 3 Calibrating the Delay Time Several delay times must be calculated one for the stop flow valve and three for the analyte collector For delay time calibrations a sample of only one of the benzoates is needed Prepare a sample of the sample concentration 0 5 mg ml dilute with 50 50 acetonitrile DO and use 50 microliter injections Set up an isocratic method of 57 acetonitrile and 43 deuterium oxide Calibrating from the LC Detector to the Microflow Probe The LC NMR
40. he Dell computer has a built in ethernet connection Establishing communication with the network at the customer site is the responsibility of the customer A network connection is highly recommended for data transfer from the PC to the Sun computer and for printing from the PC 50 LC NMR Accessory Installation 01 999036 00 C0800 Chapter 4 LC NMR Analyte Collector Sections in this chapter e 4 1 Connecting the Analyte Collector to the LC Workstation this page e 4 2 Installing the Analyte Collector Plumbing page 53 4 3 Analyte Collector Flow Diagrams page 54 4 4 Setting Up Software for Analyte Collector page 56 The optional LC NMR Analyte Collector allows the collection and storage of up to 15 analytes fractions automatically or 16 manually The Analyte Collector can be operated manually with switches on the front panel or by software with the Stop Flow Monitor program on the LC STAR workstation Designed to trap analytes as they elute from an HPLC system the Analyte Collector is capable of operating at moderate pressures and maintaining the analytes in the conditions under which they eluted When connected directly to the NMR spectrometer the Analyte Collector replaces the stop flow valve in both hydraulic and electrical connections The Analyte Collector comprises a base module and a loop module as shown in Figure 26 The base module contains two valves that control where the HPLC effluent is directed
41. he next procedure to connect the source air Connecting the Source Air This procedure describes how to connect the regulator assembly 00 990668 00 to the filter regulator assembly used by the spectrometer The new regulator is used to regulate the air used by the Powered Event Module PEM and attaches to the cross connector as shown in Figure 16 You will need a 1 4 inch Allen wrench Wrap all threads with Teflon tape before assembly LC NMR Accessory Installation 01 999036 00 C0800 Filter regulator assembly 2 6 Installing the Powered Event Module Z 5 Cross connector Bu Air supply to spectrometer Connector ala A 7 Ont New regulator assembly for PEM Air supply to PEM Figure 16 Connecting the Regulator to the Cross Connector Make sure no sample is in the probe or magnet Locate the filter regulator assembly that supplies the system on which you are installing the LC NMR Turn off the air flow to the filter regulator assembly Use a 1 4 inch Allen wrench to remove the plug from the bottom or top of the cross connector Screw the new regulator assembly into the cross connector as shown in
42. hod Error Method Metho Run Table Select Log Copy Delet Log Print Figure 18 Method Actions Display 4 With the cursor under METHOD SELECT press SELECT 36 LC NMR Accessory Installation 01 999036 00 C0800 2 8 Using the LC Pump The display shows Select Method Press Select and allows you to choose a method number We will be setting up method 1 so press the INCREASE or DECREASE button until 1 is displayed Then press SELECT The display shows Method 1 Selected Press SET CONDITIONS The display shown in Figure 19 appears Method Endtime Action Pmin Pmax Equil Time 1 6 00 Equil 0 300 0 00 Figure 19 Method 1 Conditions Display Make the values the same as shown in Table 1 Pump Method 1 Conditions Table 1 Use the CURSOR INCREASE and Field Value DECREASE buttons to change the values Endtime 6 00 After the values are set to those shown in Action Equil Table 1 press STATUS to return to the Pmin 0 status display Then go to the next Pmax 300 rocedure to create a relay program P y prog Equil Time 0 00 Creating a Relay Program This procedure describes how to create the initial relay program for testing the probe and calibrating the system If you have other devices connected to your pump such as an autosampler or analyte collector you might need to create a more detailed relay program as described in the Chromatography Systems manuals For testing the probe and calibra
43. ield enter one of the following 12 seconds if using the stop flow valve 58 LC NMR Accessory Installation 01 999036 00 C0800 5 3 Calibrating the Delay Time e 25 seconds if using the analyte collector 3 Set Hold Time to 1 minute Make sure Wait for NMR is unchecked 4 Prepare a sample that provides an easily observable NMR peak under continuous on flow conditions You should be able to see a peak with nt 4 or less 5 Set up the NMR appropriately for LC NMR Set nt 4 and set ni to a large value that allows you to observe at least one of the LC peaks that elutes off the column and through the probe Expanding the region around where you expect to see an NMR peak is helpful however you can expand this region after data is collected 6 Start the LC system Start the NMR before the peak reaches the probe 7 Monitor the NMR as the LC peak elutes through the probe As the peak moves into the probe the signal increases If your calculated Delay Time is correct you should see a profile similar to Figure 33 after you enter dssh Figure 33 Profile Showing Delay Time is Correct If the delay time is too short the profile looks like Figure 34 we LUULU ET Figure 34 Profile Showing the Delay Time is Too Short Fix this by increasing the delay time You can estimate the amount of increase required from the time required from each spectrum and the number of spectra taken from when the valve closes to when the apex passes thro
44. iners for the following reasons e To prevent dust from accumulating e To prevent excessive exposure to vapors e To prevent pickup or exchange of D with H Install a vent when the solvent container is connected to the HPLC system This is especially important for solvents that are being continuously sparged 1 6 Caring for the HPLC System and Microflow Probe This section provides recommendations about caring for the HPLC system and the Microflow probe Do not use solvents that are incompatible with PEEK quartz 316 stainless steel tefzel or teflon Also do not use solvents or buffers that are incompatible with the column s packing For example high pH buffer systems may dissolve silica in C 18 and C 8 columns Do not allow solvent to drip onto the top of the LC pump the LC detector or any other electronic device where the solvent can penetrate the cabinet and reach the circuitry If you are using buffers flush the system with DO or water daily to dissolve any salts that might have precipitated Do not leave buffers or only water in the HPLC system or Microflow probe Bacteria can grow and block tubing and the column Always rinse the HPLC system and the Microflow probe with D O or water first followed by a mixture of at least 25 organic solvent and D O or water To prevent the probe from drying out between use make sure that the probe inlet and outlet ports are sealed with a piece of tubing connected between them or with no h
45. injury or death to humans or animals or significant property damage CAUTION Cautions are used when failure to observe instructions could result in serious damage to equipment or loss of data Warning Notices Observe the following precautions during installation operation maintenance and repair of the instrument Failure to comply with these warnings or with specific warnings elsewhere in Varian manuals violates safety standards of design manufacturing and intended use of the instrument Varian assumes no liability for customer failure to comply with these precautions WARNING Persons with implanted or attached medical devices such as pacemakers and prosthetic parts must remain outside the 5 gauss perimeter from the centerline of the magnet The superconducting magnet system generates strong magnetic fields that can affect operation of some cardiac pacemakers or harm implanted or attached devices such as prosthetic parts and metal blood vessel clips and clamps Pacemaker wearers should consult the user manual provided by the pacemaker manufacturer or contact the pacemaker manufacturer to determine the effect on a specific pacemaker Pacemaker wearers should also always notify their physician and discuss the health risks of being in proximity to magnetic fields Wearers of metal prosthetics and implants should contact their physician to determine if a danger exists Refer to the manuals supplied with the magnet for the size of a ty
46. k A is the correct path At the prompt eject the disk and insert the third disk A is the correct path 3 If using the ProStar 330 install Polyview 2000 from the CD ROM This requires a superuser password Refer to Loading Polyview Software on page 24 46 LC NMR Accessory Installation 01 999036 00 C0800 3 6 Setting Up STAR 5 31 4 Disable modules that are not being used from the STAR toolbar 5 Set up the STAR software a Click on the STAR Tool bar icon left side of the screen In the right side of the tool bar at the top of the screen click the right mouse button and select Module Disable Enable from the menu that appears An upper and lower box appears All possible modules made by Varian are enabled in the top box c Highlight the module that you have not installed on your system by clicking on their names After the modules are selected click the disable button The modules that you have disabled will appear in the lower box where you can enable them later if necessary The modules that you have left enabled appear in the upper box d Click OK when you are finished 6 Set up the LC instrument from the System Control icon upper left icon on the STAR tool bar Drag the icons for the pump and detector into instrument 1 as shown in Figure 22 Double click on one of them when the icon outline turns black An image of the front panel of the module should then be displayed similar to Figu
47. l pump operations described here are selected on the front panel of the pump Refer to the Varian Chromatography Systems manual for further details on using the pump To use the STAR Chromatography software for method building see the LC NMR operation chapter in the manual User Guide Liquids NMR Creating an Initial Pump Method This procedure describes how to create a method for testing the probe and calibrating the system The method created here uses parameters that are optimized for probe testing and system calibration You can create other more intricate methods using the documentation provided by Chromatography Systems or the LC NMR operation chapter in the manual User Guide Liquids NMR A method is a group of parameters that control the pump You use the front panel keypad on the pump to create this method 1 If not already done turn on the pump power as described in the Chromatography Systems manual 2 Press the STATUS button in the OPERATION menu The display on the front of the pump shows a read only status of current pump conditions Figure 17 shows the typical initial status display for method 1 Meth SA SB SC Flow Pres Relays Time 1 100 0 0 1 00 Q 0 00 Figure 17 Status Display for Method 1 3 Press the METHOD ACTIONS button in the TABLE SELECT menu The display shown in Figure 18 appears The cursor should be under METHOD SELECT If not use the cursor buttons to move the cursor Met
48. llation 01 999036 00 C0800 Chapter 5 LC System Calibrations Sections in this chapter e 5 1 Calibrating LC1D WET this page e 5 2 13C Sideband Suppression page 58 e 5 3 Calibrating the Delay Time page 58 5 4 New Solvent Concentration page 61 The LC hardware must be installed and connected and the probe must be installed and tested before these calibrations can be done 5 1 Calibrating LC1D WET This procedure describes how to do the 1c1d WET solvent suppression calibration This procedure uses the LC1D pulse sequence and requires the 20 to 50 CH3CN D 0 sample 1 Enter 1c1d to create the parameters for the LC1D pulse sequence 2 Enter setscout ga The set scout macro turns off water suppression and lowers transmitter power The ga command acquires a nonsuppressed spectrum on the solvent mixture 3 Enter setwet The setwet macro raises the transmitter power places tof at the CH CN frequency and sets the wet shape to solvent shift for HOD 4 Enter ssfilter n to turn off solvent subtraction 5 Enter array wetpwr 20 10 0 5 ga to create an array of wet pwr in 0 5 dB steps 6 Enter dssh to display the spectra you just acquired 7 Enter wetpwr wetpwr n where n is the number of spectra with the CH3CN best suppression This calibration should be checked occasionally particularly if suppression deteriorates 8 Enter gtw 0 002 gzlv1lw 1000 2000 4000 30000 ga to run a gradient stren
49. located on the back of the console then disconnect the ac power cord WARNING Do not substitute parts or modify the instrument Any unauthorized modification could injure personnel or damage equipment and potentially terminate the warranty agreements and or service contract Written authorization approved by a Varian Inc product manager is required to implement any changes to the hardware of a Varian NMR spectrometer Maintain safety features by referring system service to a Varian service office WARNING Do not operate in the presence of flammable gases or fumes Operation with flammable gases or fumes present creates the risk of injury or death from toxic fumes explosion or fire WARNING Leave area immediately in the event of a magnet quench If the magnet dewar should quench sudden appearance of gasses from the top of the dewar leave the area immediately Sudden release of helium or nitrogen gases can rapidly displace oxygen in an enclosed space creating a possibility of asphyxiation Do not return until the oxygen level returns to normal WARNING Avoid liquid helium or nitrogen contact with any part of the body In contact with the body liquid helium and nitrogen can cause an injury similar to a burn Never place your head over the helium and nitrogen exit tubes on top of the magnet If liquid helium or nitrogen contacts the body seek immediate medical attention especially if the skin is blistered or the eyes are affecte
50. mL min 5 Press the RESET button twice 6 Press PUMP 7 After 5 minutes press STOP The solvent is transferred into the probe flow cell and the probe is ready for NMR 01 999036 00 C0800 LC NMR Accessory Installation 41 Chapter 2 HPLC Hardware Setup 42 LC NMR Accessory Installation 01 999036 00 C0800 Chapter3 Software Installation Cascade Drivers STAR 5 3 Sections in this chapter e 3 1 Loading the VEE Samples this page e 3 2 Installing Software Drivers this page e 3 3 Installing Cascade page 44 3 4 Resolving Sound Card Conflict on DELL Computers page 45 e 3 5 Reinstalling Cascade for Windows 95 98 page 45 3 6 Setting Up STAR 5 31 page 46 3 7 Setting Up the Ethernet Connection page 50 3 1 Loading the VEE Samples 1 Insert the VEE Samples disk in the A drive 2 Use Windows Explorer to look at the contents of the disk 3 Double click on the setup icon 4 Follow the instructions Note If the VEE Samples disk is missing from the shipment make a copy of the VEE samples from the LC NMR training course disk 3 2 Installing Software Drivers Driver Installation The DT2801 driver must be installed so that system can recognize the Data Translations ADC board 1 Insert the floppy disk labeled DT2801 Device Driver for win3 11 and win95 Go to the Control Panel and select Add New Hardware Select Next Answer No to Do you want windows to search and click
51. ntage of organic solvent may not allow a visible amount of solvent to accumulate because of evaporation If no visible leak is seen then the pump head may be leaking internally piston seal or inlet valve or if included in the test the injector valve might be leaking internally If the constant pressure leak test produces a higher leak rate than is tolerable after running it for 10 minutes and no visible leaks can be found run the pressure decay leak test If the results of this test are acceptable then the fittings between the outlet check valve and the plug are leak tight and an internal leak in the pump head is indicated Check carefully around the drain passage at the bottom of the pump head If no leakage appears here then the inlet valve may be leaking since leaks here can not be seen externally If the results of the tests are not conclusive the leak tests should be repeated before changing any of the hardware in the pump head 01 999036 00 C0800 LC NMR Accessory Installation 35 Chapter 2 HPLC Hardware Setup 2 8 Using the LC Pump This section describes how to use the 9012 Solvent Delivery System LC pump from the front panel keypad for the following operations e Creating an Initial Pump Method e Creating a Relay Program e Priming the Pump e Flushing the Probe Flow Cell e Purging the Transfer Tube e Transferring a Solvent into the Probe You will use these procedures when you are installing and testing the probe Al
52. o NMR violet 9 Error NO yellow 10 Error C jumpered from pin 18 11 Step Input white red 12 Step Input white black white brown 13 BCD 1 1 output brown 14 BCD 1 2 output red 15 BCD 1 4 output orange 16 BCD 1 8 output green 17 BCD 10 1 output blue 18 Common black 52 LC NMR Accessory Installation 01 999036 00 C0800 4 2 Installing the Analyte Collector Plumbing 4 2 Installing the Analyte Collector Plumbing The Analyte Collector replaces the stop flow valve in the LC NMR system The procedure below describes how to install the plumbing for the base module of the Analyte Collector Figure 27 is a diagram of the plumbing connections 1 SO a ON ne ee a 10 Turn off the LC pump LC detector and exit from the Stop Flow Monitor program Connect the outlet from the mixer to the From Pump connector Connect the inlet to the injector valve to the To Column connector Connect the outlet of the column to the From Column connector Connect the inlet of the detector to the To Detector connector Connect the outlet of the detector to the From Detector connector Connect the NMR transfer line to the To NMR connector Connect the To Waste connector to an approved waste container For stopped flow or on line runs connect a tube between Collector In and Collector Out Do not connect the loop module For analyte collection or analysis connect the loop module to the appropriate Collector In and Collector
53. ole ferrules connected to each port Similarly seal the 310 detector if it will not be used for a while Periodically flush the probe with the strong solvent you are using to remove any compounds that may adhere to the probe flow cell Do not connect tubing with less than a 0 010 inch internal diameter I D to the probe outlet larger I D s are preferred Long runs of small I D tubing cause excessive pressure in the probe that can break seals and create leaks within the probe Do not subject the probe to excessive mechanical vibrations or violent motion 18 LC NMR Accessory Installation 01 999036 00 C0800 Chapter 2 HPLC Hardware Setup Sections in this chapter e 2 1 Assembling the Valves and ProStar LC Pump Plumbing this page e 2 2 Setting Up the ProStar LC Detector page 21 e 2 3 Installing the Stop Flow Valve and Plumbing page 25 e 2 4 Unpacking and Assembling the LC STAR Workstation page 27 e 2 5 Connecting the Stop Flow Cable page 28 e 2 6 Installing the Powered Event Module page 31 e 2 7 Leak Testing the System page 34 e 2 8 Using the LC Pump page 36 Most of the HPLC hardware is set up using the instructions provided in Varian Chromatography Systems documentation Detailed instructions are provided in this chapter for plumbing the pump and valves connecting the detector assembling the LC STAR workstation and connecting the HPLC system to the NMR spectrometer The NMR spect
54. on the 310 detector and connect the end labeled P10 310 to J10 on the LC pump 330 detector connect the cable labeled P10 to J10 on the LC pump Connect the cable labeled P9 to the J9 on the LC pump c If an analyte collector is not used connect the cable labeled P5 to the connector on the REMOTE SWITCHING cable of the stop flow valve If an analyte collector is used do not connect P5 5 Make sure the thumbwheel switch S201 DEVICE ADDRESS on the back of the LC pump is set to 1 if not set it to 1 6 Make sure the thumbwheel switch S201 DEVICE ADDRESS on the back of the LC detector is set to 3 if not set it to 3 The stop flow cable is now connected 2 6 Installing the Powered Event Module This procedure describes how to install the air line used by the Powered Event Module PEM shown in Figure 14 This procedure also describes how to connect the PEM to the other LC hardware The two procedures in this section describe how to connect the data cables and install the air line into the source filter regulator air line The PEM requires a clean air nitrogen source of 60 psi minimum pressure Connecting the Injector Valve and the Pump This procedure describes how to install the air lines to the injection valve and how to connect the data cable between the Powered Event Module and the Detector Leave the power cord disconnected and the power switch off until all other connections are made 01 999036 00 C0800 LC NMR Acce
55. opped to zero remove the plug and reconnect the solvent line Turn off the power switch and then turn it back on to restore normal operation of the 9012 LC pump Interpreting Leak Test Results The constant pressure leak test displays the leak rate directly in L min Because of the extreme sensitivity of this test take care in interpreting the reported leak rate Relate the leak rate to the operations flow rate to determine the significance of a reported value For example a leak rate of less than 1 wL min will be unimportant at a flow rate of 1mL min The pressure decay mode displays a pressure drop and an elapsed time If the hydraulics being tested include the standard damper the leak rate in L min may be estimated by multiplying the pressure drop in atm by 1uL atm and dividing by the elapsed time Both test modes are sensitive to fluctuations in ambient temperature which can lead to thermal expansion or contraction of the liquid in a closed system This can cause small fluctuations in the pressure or leak rate readings but if a leak exists it should become apparent after the system has been under pressure for 5 or 10 minutes As a rule of thumb a drop of water is approximately 50 uL For organic solvents or mixtures of organic and water a drop is typically smaller than 50 uL Therefore with a leak rate exceeding 25 uL min liquid should be visible at one of the fittings after several minutes of test Lower leak rates with a high perce
56. ot Carefully align the board connector with the connector inside the PC the rear panel connector on the board must face the rear of the PC Then using your thumbs gently push the board into the connector until the board is fully seated When correctly seated the 50 pin ribbon cable connector on the ADC board should be accessible through the back of the PC Replace the cover on the PC The ADC board is now installed 01 999036 00 C0800 LC NMR Accessory Installation 27 Chapter 2 HPLC Hardware Setup 2 5 Connecting the Stop Flow Cable This procedure describes how to connect the stop flow cable between the PC NMR console pump stop flow valve and the detector The stop flow cable begins at the interface box at the 25 pin D shell connector The harness then divides into cables that connect the LC with the NMR console and cables that connect to the LC hardware The PC connects to the interface box with a ribbon cable from the ADC board e Figure 9 shows a diagram for connecting the stop flow cable to the 310 detector NMR console LC STAR workstation interface box 310 LC Detector 2 5A SB ATTENTION tara WIRED FOR 120 VAC OPERATION RELAYS Do not connect P5 if using the analyte collector 230 LC pump a PRESSURE J2 Saimmv
57. phy 11 high power amplifiers cautions 10 high pressure flow cell 24 HPLC capabilities 11 hardware components 19 system care 18 I injector valve connection 31 inlet ports 38 installation planning 13 installation requirements 13 interface box 28 30 L LC detector setup 21 thumbwheel switch 31 LC pump creating a method 36 creating a relay program 37 inlet ports 38 method 1 conditions 37 priming 38 priming valve 38 pumping solvent to probe 40 relay program outline 38 thumbwheel switch 31 LC STAR workstation 14 LC Stop Flow Monitor program installation 44 LC system calibrations 57 LC workstation ADC board installation 27 LC NMR Accessory Installation 63 Index installing drivers 44 lc1d WET solvent suppression calibration 57 LC NMR accessory compatibility 12 components 11 electrical outlets requirements 13 HPLC hardware set up 19 lab layout guidelines 14 new solvent concentration 61 preinstallation requirements 13 leak test 34 35 liquid chromatographer 13 M magnet quench warning 8 magnetic fringe fields 14 magnetic media caution 9 metal objects warning 7 method building with STAR 5 3 48 method creation on pump 36 method defined 36 N National Instruments CD 24 nitrogen contact with body 8 nitrogen gas flowmeters caution 10 NMR spectroscopist 13 nonmagnetic table 13 P pacemaker warning 7 PCI GPIB board 24 PEEK tubing 19 colors 19 compatibility 18 PEM Powered Event Module 31 32
58. pical 5 gauss stray field This gauss level should be checked after the magnet is installed WARNING Keep metal objects outside the 10 gauss perimeter from the centerline of the magnet The strong magnetic field surrounding the magnet attracts objects containing steel iron or other ferromagnetic materials which includes most ordinary tools electronic equipment compressed gas cylinders steel chairs and steel carts Unless restrained such objects can suddenly fly towards the magnet causing possible personal injury and extensive damage to the probe dewar and superconducting solenoid The greater the mass of the object the more the magnet attracts the object Only nonferromagnetic materials plastics aluminum wood nonmagnetic stainless steel etc should be used in the area around the magnet If an object is stuck to the magnet surface and cannot easily be removed by hand contact Varian service for assistance 01 999036 00 C0800 LC NMR Accessory Installation 7 SAFETY PRECAUTIONS Warning Notices continued 8 Refer to the manuals supplied with the magnet for the size of a typical 10 gauss stray field This gauss level should be checked after the magnet is installed WARNING Only qualified maintenance personnel shall remove equipment covers or make internal adjustments Dangerous high voltages that can kill or injure exist inside the instrument Before working inside a cabinet turn off the main system power switch
59. quirements 13 Acquisition Controller board 31 ADC board 27 28 Analog Output board 23 Analyte Collector 51 flow diagrams 54 automatic teller machine ATM cards caution 9 B bacteria growth prevention 18 blue PEEK tubing 19 Breakout panel 30 buffers use 18 C cable lengths 14 calibrations 57 caring for the HPLC 18 Cascade software initializing 45 installation 44 reinstalling for Windows 95 45 cautions defined 7 chromatography 11 Chromatography Systems documentation 19 column packing compatiblity 18 compatibility of LC NMR accessory 12 constant pressure mode 34 contamination prevention 39 credit cards caution 9 cross connector 32 cryogen servicing requirements 14 D D20 requirements 13 DATA SYSTEM OUTPUT 31 Data Translations ADC board 43 DELL computer sound card conflict 45 detector 28 J2 connector wiring 21 ProStar 310 21 ProStar 330 22 thumbwheel switch 21 DT2801 Device Driver For Windows 43 DT2801 driver installation 43 DTM ADC board DT2801 driver installation 43 01 999036 00 C0800 Index E Ext Time Base 31 external event relay 37 F filter regulator assembly 31 flammable gases warning 8 flow cell flushing 39 flow cell high pressure 24 flushing the probe flow cell 39 G getwet macro 6l GPIB address 24 GPIB board 24 H helium contact with body 8 helium gas flowmeters caution 10 helium requirements 13 Hg lamp 22 high pH buffer systems 18 high performance liquid chromatogra
60. r Method End Action Pressure Limits Equilibration Time Sgh ProStar 330 PDA Detector Address 71 End Time rin nooo Miom o E Gk ProStar 330 PDA Detector Control 0 00 min woe Breen Action Equilibrete E Maximum p50 el PolWiew E Peaksense Time Program Hf Commands Method solvent Description JE Reports Method Eih Channel 1280 60 nm A ACN EM Data Handling BB tegration Parameters Peak Table Calibration Setup i elie Ready O Nm B Deuterium Oxide Le Le Le MeoH Figure 24 STAR Pump Method Window 48 LC NMR Accessory Installation 01 999036 00 C0800 3 6 Setting Up STAR 5 31 2 Set up the pump method as needed Figure 24 shows an example of a gradient method 3 Move to the detector module click on Method Parameters Figure 25showsan ProStar 310 Detector Method Window examples of ProStar 310 and 3 30 methods E ProMethel mth Method Notes 1 Prostar 230 SOM Control Address 1 amp Method Parameters 4 To set the display Prostar 310 UV Vis Detector Address Gr ProStar 310 Vis Detector Contro wavelength in the A gene jata Handling ProStar 330 go to a libration Setup the peak sense Fae time program option of Polyview Ed cattraton Slack Repor For Bh ProStar 330 PDA Detector Address 71 2000 select the a desired wavelength This should update the pekte TA Calibration Setup moe a channel 1 heading 5 To convert results files to
61. r a name and click on Save Open the File menu again and select Exit Setting Up Method Parameters 1 Click on method parameters under the 9012 or ProStar230 as shown in Figure 24 Dota s s ES ProMethd mth Method Notes amp ProStar 230 SDM Control Address 1 Method Parameters f ProStar 310 UV Vis Detector Address Gf ProStar 310 UV Vis Detector Contro SESLE gt BS Method Builder ProMethd mth _ I Se Ele Edit View Window Help el x ts alele aae exe ale a gt gt i ee E SEI a B Probethdimth i A fa P Eoss N Time Events Program Sei oh tea ree hese aan SS xa ye e Pee Rela Rela Rea a meters malfmnin f eak A hos da Prost Vie Detector Adaressl 10 5050 oot lor wor be Of ie er amp Proster 310 UV Vis Detector Contrg 8 Method Parameters eG Channel1 280nm__ EH Dats Handling E integration Parameters E Peak Table Calibration Setup Verification Setup 0 01 1050 80 00n lon Jor mor Insert 1 00 10 50 50 oon Selon mhor mor alelelelela lejeje KOKNKOKOKIEI ec foo fs fen fon fa fes fro Time Events Table Fr P Sa Selectisn EK Standard Reports I ree 2 De E Javailable Table Entries 66 Tila 489 Chromatogram Format Calibration Block Report Fo
62. r to the Cross Connector e ccc eeceseeecseeesecseeesecnseeeeneeenees 33 Figure 17 Status Display for Method 1 eee cesses eeseeseceeeeseceeeeeeeesevenecaeesaecsassaecnessaeeeeenees 36 Figure 18 Method Actions Display secesecseceseeseceseeseceeseseceeeececaeveeecaeesaecaeesaecneesaeeeesaees 36 Figure 19 Method 1 Conditions Display cece ceeeeeeseeseceeeeseceeceeeecaeeeeecoeesaesaeesaecnessaeeseeaees 37 Figure 20 Relay Program Display x sesicssccssisessacsaanscoptsessscusdsSesnesbaisbanessnebiveceeaiesespacebatyieseseaseensdhbeabe 37 Figure 21 Prime Valve and Drain on Front of LC Pump uu ee eeeeeceecneeseeeeceeceeesceaeceeeeesaeaeeees 39 Figure 22 STAR Configuration Window 0 cceseecsesssescsecseeeceecsseceseceecseeseesecnecaeesesaecaeeeesaeeaseees 47 Figure 23 STAR System Control Window ccceceeseeeeseeesceeeeeeceeeeeeecseeeeecaeesaecaaesaecessaeeneensees 47 Figure 24 STAR Pump Method Window qu cee seeeseeseceseeseceeeeseceseeeecaeeeecoeesaecseesaecnessaeeeensees 48 Figure 25 Detector Method WindowS sesesecseceseeseceseeseceeeeseceeseseeeaeeeecaeesaecaaesaecnessaeeaeenaees 49 Fisure 26 LC NMR Analyte Collector scsi csisce satcncss avsessnsvacsccsasdvsctansossvnatesteesesathuucrsabessavaenvaevoane 52 Figure 27 Analyte Collector Hydraulic Connections sesssseesssessereseeeserrsresrsreerereserrnseseesenens 53 Figure 28 Flow Diagram for Collecting Separated Peaks e eee eceseeecseeeseceeeese
63. re 23 Figure 23 STAR System Control Window 01 999036 00 C0800 LC NMR Accessory Installation 47 Chapter 3 Software Installation Cascade Drivers STAR 5 3 Building a Method in STAR 5 31 This procedure describes building a method in STAR 5 31 1 Enlarge the pump and detector windows to monitor the pump and detector states 2 Click on the method builder icon second from the left in the STAR toolbar similar to Figure 22 Choose create a new method A wizard pops up 3 Choose Next In the new window that pops up select a configuration for instrument 1 Both the pump 9010 9012 or ProStar230 and detector 9050 9065 ProStar3 10 or ProStar330 modules should be listed on the right hand side 4 Click Next Another window pops up Make sure that the correct detector is listed 5 Click on Next A window pops up listing sections for post run processing Choose both data handling and standard reports 6 Click on Next A final window pops up It should list the pump detector and both data handling and standard reports 7 Click on Finish A window pops up asking you to choose between opening a method or creating a new method 8 Click on Cancel The Method Builder screen with all the sections and subsections is displayed Follow the instructions in Setting Up Method Parameters on page 48 to set up the method parameters 9 When finished setting up the method open the File menu and choose Save As Ente
64. rometer must be installed and running properly before the HPLC system can be integrated with it This chapter also provides some basic instructions for using the LC pump and detector If you are unfamiliar with chromatographs and with Varian STAR Chromatography systems in particular you may wish to read the Varian Chromatography Systems manuals before beginning the HPLC hardware setup The installation kit provides two rolls of PEEK tubing The blue PEEK tubing 0 010 inch I D is used to connect the various LC valves The red PEEK tubing 0 005 inch I D is used as the transfer line to connect the probe to the LC system 2 1 Assembling the Valves and ProStar LC Pump Plumbing The injector valve purge valve column valve mounting plate mounting track column mounting hardware and thumbscrews are packaged in separate plastic bags in the universal valve mounting kit This kit enables you to mount these valves and the column onto the side of the LC pump which has holes tapped in both sides We suggest you mount the assembly on the left side of the pump to facilitate plumbing connections Figure 2 illustrates how the universal valve mounting kit is assembled Instructions in the Chromatography Systems manuals provide further information 01 999036 00 C0800 LC NMR Accessory Installation 19 Chapter 2 HPLC Hardware Setup Universal valve mounting kit Inject stem Injector Valve mounting valve plate naai h E p
65. s 2 3 Installing the Stop Flow Valve and Plumbing ooo eee eee cseesee ce ceee ce ceseeeeeeeerees Unpack and Connect Plumbing oo eee eeeesecsseeeeceeeeeceeeseeeeeeeeseneeaeeeaes Prepare the Transfer Tube raiione tin e nE S cece AEAT EEA TESNE 2 4 Unpacking and Assembling the LC STAR Workstation esesesseeseereeseerereererrsreeesrees 2 5 Connecting the Stop Flow Cable oo eee ceessecsecneecnecsseeseceseeseeeeseseseeeeaseaeseeeeaee 2 6 Installing the Powered Event Module oo eee cece ceceseeeeceseeeeeeseeeeeeaeeeaeceeeaeenaes Connecting the Injector Valve and the Pump ow eee cece ceeeeeeeeseeeeeeeeeeneenee Connecting the Source Air sisssesessercesorcrsererisessssevoseisossesosessosesesesseresseresso esors 2 7 Leak Testing the System lt cicicescsssiscebscceck asevsacsbves btiecseessseasebecacedereoniezsselue se caousagecsees Running the Leak Test vice hah i do eee A ee Interpreting Leak Test Results oo eee eeeecsseeeneeceeeesneceeeeeeeeeeseeceeeceeeeeneecneeees 2 6 Using the LC Pump ssiccienn nie nue nah EEEE ain A A Creating an Initial Pump Method oo cee ceeeeeceeeeeceeceeeseeeeeeseeeeeetenes Creating a Relay Program cite csati seit areare ere EEOAE EE EREE ETES Priming the PUMP a eesceesecsseeeseeceecececeeeeeesecesaecseecsaeeeaeeceeeeeaeceeneeeeeesaeeees Flushing the Probe Flow Cell oi ecceicc ei ceessecsecsseceeceseeseceseeeseeeeeeeseaeseeeeneeeaeenee Purging the Transfer Tube oo eecceceeesseeeceeeseeeseeceeeecnaeceaeeceeee
66. screw the thumbscrews from the Hg lamp Remove the cover with the lamp and remove the sponge from the detector as shown in Figure 6 22 LC NMR Accessory Installation 01 999036 00 C0800 2 2 Setting Up the ProStar LC Detector Transport Sponge a Wi Figure 6 ProStar 330 Sponge Removal 5 If the modules are to be stacked the ProStar 330 should be placed on top of the ProStar 230 plump Installing the Analog Output Board Figure 7 shows the location of the Analog Output board Connect to pump start out or relay output Analog Output board RELAY BOX RECORDER RANGE ic x ic x 10 X 100 Connect to GPIB on PC RECORDER RANGE co NORMAL o Low PROCESSOR Set address to 9 FUSE T3 15AL 250V fr ADDRESS z L LJ Figure 7 ProStar 330 Analog Output Board Install 1 Remove the blank panel from the left side on the back of the ProStar 330 by loosening the screws 2 Install the Analog Output board 3 Set the switch to normal for calibration purposes The switch can be set to low for better detection of small peaks The x1 x10 and x100 settings are only used with the recorder output which will not be accessed 01 999036 00 C0800 LC NMR Accessory Installation 23 Chapter 2 HPLC Hardware Setup Installing the High Pressure Flow Cell Replace the low pressure flow cell with the high pr
67. ssory Installation 31 Chapter 2 HPLC Hardware Setup lt 32 gt POWER J405 J406 a g GAS 200 VA Max eo IN OUT wired for TaD INC N O N C J401 Q 402 shos J404 i green pink Air in from ba lace J404 connects to RELAY 1 J3 on the pump To injector valve Figure 14 Powered Event Module Back Panel Connectors 1 Connect a cable from J404 on the back of the PEM to the connector on the back of the LC pump labeled RELAY 1 or J3 2 Connect one end of the long source air tube 1 8 inch O D to the hose barb labeled IN see Figure 14 Leave the other end of this tube disconnected until the next procedure 3 Make the air connections between the PEM to the injector valve as follows a Locate the two hose barbs on the side of the injector valve see Figure 15 b Connect one end of the pink 1 8 inch O D tube to the hose barb labeled N C Connect the other end of this tube to the hose barb on the injector valve that is farthest from the mounting plate see Figure 15 Mounting Hose barbs Pelee Figure 15 Injector Valve Top View c Connect one end of the green 1 8 inch O D tube to the hose barb labeled N O Connect the other end of this tube to the hose barb on the injector valve that is closest to the mounting plate see Figure 15 4 Go to t
68. st 100 mL of acetone dg At least 300 mL of OmniSolv brand acetonitrile EM Science part no AX0142 1 e Research grade sucrose e At least three 100 mL volumetric flasks for preparing standard samples e An appropriate waste container for HPLC effluent The cover of the container must have a hole in the top of about 2 mm diameter The hole is used for the outlet of the LC NMR Microflow probe e Air supply for the pneumatic valve Site Your site must have the following e Five 110 Vac or 220 Vac outlets within 2m 6 ft of where the table holding the LC hardware will be located Several extension cords should be available for the installation Approval by Varian of the proposed layout of the LC NMR lab Personnel A person experienced in liquid chromatography must be present during the installation Having this person on site ensures that vital information is shared regarding the following e The performance of the HPLC system e The interface between the LC system and the NMR spectrometer e The isolation of the LC system from the NMR spectrometer e The overall operation of the LC NMR accessory At least one NMR spectroscopist must be present during the installation calibration and system tests 01 999036 00 C0800 LC NMR Accessory Installation 13 Chapter 1 Introduction 1 4 Site Layout Guidelines This section provides guidelines for arranging an LC NMR lab The illustrations in this section provide a room layout
69. system can perform both continuous on flow spectral measurements and stop flow measurements Continuous on flow experiments do not require a delay time A stop flow measurement is typically used if the sample concentration is too low to be seen on flow or if more extensive experiments such as COSY TOCSY or HMQC are required In order to do stop flow a detector more sensitive than the NMR is required to allow the LC peak of interest to be detected and stopped in the NMR probe Between the detector and the NMR probe is a volume that is equal to the tubing connecting the two detectors any intervening devices and the volumes of the flow cells of the detector and the NMR probe Since the LC system is forcing solvent through the system at a flow rate set by the operator an LC peak requires a finite amount of time to travel from the detector to the NMR probe This time called the delay time is calculated with the following procedure A typical delay time is 10 12 seconds using the stop flow valve at a flow rate of 1 mL min with 3 m 10 ft of 0 005 in I D tubing connecting the stop flow valve to the flow probe If the analyte collector is plumbed in the delay time is on the order of 25 29 seconds at 1 mL min 1 Start the LC system and begin the Cascade LC NMR program on the PC If using p hydroxy methyl benzoate set up an LC method for 50 acetonitrile 50 D20 ImL min flowrate 5 minute run time and 280 nm detection 2 Inthe To NMR Probe f
70. t Ths Mantal smerenie ee eee rE a EEEE EEREN ENRE NE ria 12 Compatibility isssisscpeneter eek ees earet eerie ts EEn sada EE EKE EE EEEn E r ESE E inei trei 1 3 Preinstallation Requirements sessseeeseseeeesreresrsesserersrresteerereereseereresserresereeseeereseereres Equipment Samples and Solvents essesseseeeeesseesssresrsrssersesreesrerrsresrrrrssesrsse DUS A E A E A A a E asta vasheceinee Pers nn l siosurpenp ura e E E A E E AET LA Site Layout Guidelines sui scscisesssscssisssesescescsseesuceicubiecvecgubevssbececasecseocbssacbece a aioe LC NMR Minimum Room Layout eesessessssssesessreserrererrssrrrssrerrsreererererreresresen LC NMR Suggested Room Layout 1 sseessssessseerssrereeresreresrerrsrerrsrrnrrrrnrreresen LC NMR Suggested Room Layout 2 esesessssseessrerssrereeresrerssrerrsrerrsrrererreeeerseee 1 5 Prepating Solvents svosccsascisasessasisaceeaseseesheeusesespsseiacsyadh EE EEEE E EET EE E ESE EEEE S 1 6 Caring for the HPLC System and Microflow Probe ssssseseseeeeeesreeerssrsrereesrrerereesrees Chapter 2 HPLC Hardware Setup csssseccssseeceeeesseeeeseessneeeeesenneneeensneeees 2 1 Assembling the Valves and ProStar LC Pump Plumbing ou eee eee eeeeeee eee 2 2 Setting Up the ProStar LC Detector oo eee ceeeceeceseeeeceseeeeeeseseeeeseeeaeceeeaeenaes Setting Up the ProStar 310 Detector oo eceeeeeeeeeeeseceeeeaecnseeaeenaeeaees Setting Up the ProStar 330 Detector oo elec eseseeeeceseeeeecaeecaecnseeaeenseeaee
71. ter provided to cut the tubing to the desired lengths A 12 foot length of red PEEK tubing will be used as the transfer line between the 4 port on the Stop Flow valve and the IN port on the probe This tubing can be shortened to reduce the delay time The 9012 LC pump ships with stainless steel tubing You may use the stainless steel tubing instead of the PEEK tubing for connections between the LC pump mixer and purge valve All connections between the 310 LC detector column and probe must use the PEEK tubing Unpack and Connect Plumbing Unpack the stop flow valve and place it on the table or cart near the injector and purge valve 01 999036 00 C0800 LC NMR Accessory Installation 25 Chapter 2 HPLC Hardware Setup 1 Connect a tube from the bottom port on the front of the pump to the large volume mixer Part No 03 919622 01 2 Connect a tube from the other end of the mixer to the 2 port on the stop flow valve 3 Connect a tube from the 5 port on the stop flow valve to the top port on the front of the detector 4 Connect a tube from the 3 port on the stop flow valve to one of the high pressure ports on the purge valve Use the SSI ferrules provided in the kit to connect the tube to the high pressure port Prepare the Transfer Tube Prepare the transfer tube which is used to transfer solvents from the LC pump to the probe 1 Find the 12 foot red PEEK tubing from the installation kit 2 Connect one end of the transfer
72. the National Instruments CD from the drive and shut down the computer After the computer has shut down turn off the power and remove the power cord Remove the computer cover and install the GPIB board in one of the PCI slots as shown in the National Instrument Getting Started manual After the board is installed replace the covers connect the power and turn the PC on Windows should briefly display a New Hardware Found dialog box If it does not refer to the National Instruments manual for a way to manually force windows to find the GPIB board Attach the GPIB cable to the PCI GPIB board and then to the GPIB connector on the back of the ProStar 330 Do not connect this GPIB cable to any other ProStar module Set the GPIB address on the rear of the ProStar 330 to 9 Loading Polyview Software 1 Insert the Star 5 3 CD e If the STAR installation wizard pops up select Install and the select Polyview 2000 24 LC NMR Accessory Installation 01 999036 00 C0800 2 3 Installing the Stop Flow Valve and Plumbing e If the wizard does not pop up use Windows Explorer to find the setup exe file in the Polyview folders on the STAR 5 3 CD 2 Click on Next on the welcome screen 3 Enter name and company information in the user information screen For the password use the password found on the yellow sheet in the Polyview Communications kit Note This password is different than the STAR 5 3 password 4 Click Next on the soft
73. ting the system you only need to create a program for relay 1 which controls the injector valve Relays can be time programmed to control six external event relays You will use the front panel keypad on the pump to create this relay program 1 Press the RELAYS button in the TABLE SELECT menu The relay display for method 1 appears as shown in Table 20 The dashes under Relays On represent the six external event relays on the back of the pump Method Line Time Relays On 1 1 O00 o ase Figure 20 Relay Program Display Press NEXT LINE in the EDIT menu If the system says that no line exists press INSERT LINE to add a new line Line 2 is created Notice that the time has changed from 0 00 to 0 01 Use the CURSOR buttons to move the cursor under the first dash under Relays On This represents relay 1 which is connected to the injector valve Press INCREASE to turn on relay 1 Press NEXT LINE in the EDIT menu If the system says that no line exists press INSERT LINE to add a new line 01 999036 00 C0800 LC NMR Accessory Installation 37 Chapter 2 HPLC Hardware Setup 38 Line 3 is created 5 Move the cursor to the Time field and push the INCREASE button until the Hable 2 Tital Belay Progen ios Pump Method 1 time is 1 00 6 Move the cursor to the first relay under Method Line Time Relays On the Relays On field and push 1 l 000 ees INCREASE to turn off relay 1 1 2 0 01 l Th
74. tinue 4 After the NMR and Cascade finish change the from loop to probe delay time to 11 seconds in the System Configuration menu 5 Inthe Analyte Collector window set the first and last loop to be analyzed to 2 6 When the LC pump and detector are ready click Start Run Loop 2 will be emptied and the new delay time will be invoked after a peak is detected 7 Increment the from loop to probe delay time for each subsequent loop and set up an analyte analysis run for that loop The correct delay time is the one that results in NMR data as shown in Figure 33 Typical values are between 10 and 15 seconds at 1 mL min Note Changes in the length or I D of the connecting LC tubing will require the delay times to be recalibrated Changes in flowrate will also affect the delay time but new delay times can be calculated For example changing the flowrate by half results in a two fold increase in the delay time 5 4 New Solvent Concentration For a new solvent concentration enter the macro get wet The get wet macro runs setscout ga setwet ga which acquires an nt 1 scout scan and a solvent suppressed spectrum of the new solvent mixture 01 999036 00 C0800 LC NMR Accessory Installation 61 Chapter 5 LC System Calibrations 62 LC NMR Accessory Installation 01 999036 00 C0800 Numerics 13C sideband suppression 58 25 pin D shell connector 28 9012 Solvent Delivery System pump 36 A acetone d6 requirements 13 acetonitrile re
75. ugh the probe If the delay time is too long the profile looks like Figure 35 EEE Figure 35 Profile Showing the Delay Time is Too Long Fix this by decreasing the delay time 01 999036 00 C0800 LC NMR Accessory Installation 59 Chapter 5 LC System Calibrations 60 Calibrating from the LC Detector to the Analyte Collector The NMR system is not needed for calibrating from the LC detector to the analyte collector It is best to use an isocratic run Make sure that the end time of the 330 or 310 detectors matches the end time of the 230 pump 1 Open the System Configuration window of the Cascade program Make sure that the Analyte collector on line box is selected Select Analyte Collector for the Experiment Type On Flow Stop Flow Make sure Wait for NMR Complete is not selected Enter 5 seconds for To Collector Loop Check to see that the calibration is starting in loop 1 Make sure the Inhibit apex detection is not selected 2 Start a run with the benzoate sample use a 50 uL injection equal to 25 mg Use a 50 acetonitrile 50 DO method with 1mL min flow rate Set the detection wavelength to 280 nm The first sample will be stored in loop 1 with the first loop fill delay time of 5 seconds 3 Repeat steps 1 and 2 using different delay times in one sec increments up to 11 seconds 4 Store successive samples in successive loops This process fills 7 loops Record the apex intensity of the sign
76. uipment LC computer LC STAR compressed gas cylinders steel chairs and steel carts away from the magnet Unless restrained such objects can suddenly fly towards the magnet causing personal injury and extensive damage to the probe the dewar and the superconducting solenoid Only nonferromagnetic materials such as plastics aluminum wood and stainless steel should be used in the area around the magnet dewar NMR Console Detector Magnet 12 6 2x Stop Flow sValve 3d Figure 1 LC NMR Block Diagram LC NMR Accessory Installation 01 999036 00 C0800 1 3 Preinstallation Requirements 1 3 Preinstallation Requirements This section lists the items needed for the LC NMR accessory installation Everything listed in this section must be in place before you begin the installation Equipment Samples and Solvents You must have the following equipment samples and solvents e A nonmagnetic table at least 0 9 m by 1 2 m 3 ft by 4 ft to hold the LC hardware e Either gas cylinder of helium outfitted with 0 25 inch OD Teflon tubing with aerator this will be used for sparging the acetonitrile solvent or the necessary degassing equipment Do not use an aspirator At least 300 mL of D O 99 8 atom percent or better e Atlea
77. urge prime Control knob valve valve shaft Jam nut D Q Weld Thumbscrew nut Column mounting hardware te Column mounting _7 bracket Mounting Attach to side track of LC pump Column gt retainer bracket Column Figure 2 Mounting the Injector and Purge Prime Valves 20 LC NMR Accessory Installation 01 999036 00 C0800 2 2 Setting Up the ProStar LC Detector 2 2 Setting Up the ProStar LC Detector Use the instructions provided in the Varian Chromatography Systems manuals to unpack and set up the ProStar LC detector Choose one of the procedures below to set up the ProStar 310 detector or the ProStar 330 detector Setting Up the ProStar 310 Detector Set up the ProStar 310 detector as follows 1 Connect a cable from J201 on the detector to J201 on the pump 2 Make sure the J2 voltage switch on the back of the detector is set to 1 V as shown in Figure 3 3 Make sure the thumbwheel switch S201 DEVICE ADDR on the back of the LC detector is set to 3 if not set it to 3 as shown in Figure 3 J2 voltage switch 310 LC Detector back panel m POWER IN Fos AY a 69 oe ey lt a ATTENTION RELAY J WIRED FOR G JEEE 488 J as Thumbwheel switch ae GPIB IEEE 488 201 DEVICE ADDR Figure 3 310 Detector Voltage and Device Address Switches
78. valve with acquisition of data by the NMR system Installing the Cascade program on the LC STAR workstation running Microsoft Windows 98 involves loading and initializing the software Loading the Cascade Software 1 2 3 4 mn Insert disk 1 into the floppy drive Click on Start select Settings and open the Control Panel Select Add Remove Programs Click on the Install button and follow the directions on the screen Windows should start running setup exe from the floppy Insert disk 2 when asked to do so Restart the computer 44 LC NMR Accessory Installation 01 999036 00 C0800 3 4 Resolving Sound Card Conflict on DELL Computers You should see a message about VTDAD 386 during the startup process Initializing Cascade 1 O ne A Click on Start select Programs and launch Cascade After Cascade starts a window pops up with the status of the A D board Click OK Pull down the Setup menu and select Communications Set the loop increment delay to 50 and the loop read delay to 10 000 Change other parameters such as run time as necessary Close the Configuration window 3 4 Resolving Sound Card Conflict on DELL Computers A memory conflict between the onboard sound card in the DELL computer and the DT2801 Data Translation might exist The conflict appears as a yellow exclamation point next to one of the drivers in the Control Panel System Manager Fix the conflict as follows 1 SO OR ra ON O Se
79. ware license agreement window 5 Click Next on the information window 6 Another window appears which offers the option of selecting a file folder for Polyview 2000 Use the default setting 7 A final window appears after the software is loaded saying the setup is complete Click on Finish Turning the Lamp On and Off 1 Before turning the lamp on right after an installation the LC system must be set up in the instrument configuration window of Star 5 3 Refer to the next chapter for instructions 2 Once the instrument is configured the lamp is turned on from Star 5 3 software by clicking the lamp on button on the ProStar 330 panel in System Control in Star 5 3 If the PC has recently restarted the ProStar 330 software must be downloaded before it is possible to turn on the lamp 3 The lamp on the ProStar 330 should be shut off from STAR software Turning off the ProStar 330 by using the switch on the front panel requires the downloading of the software control code when the ProStar 330 is turned on again This takes several minutes 2 3 Installing the Stop Flow Valve and Plumbing The procedure in this section describes how to install the stop flow valve and the plumbing If you have an Analyte Collector with your system skip to HPLC Hardware Setup on page 19 the Analyte Collector replaces the stop flow valve For this procedure use the 0 010 inch I D blue striped tubing to make the connections Use the tubing cut
80. ys included in the closed system Operation of the leak test without the pressure transducer in line could result in damage to the pump 4 On the 9012 LC pump press and hold the SELECT key while simultaneously turning on the power This will display the Extended Diagnostics Menu 5 Scroll to the Leak Test using the NEXT LINE key Press START 6 Select either Constant Pressure or Pressure Decay mode using the INCREASE or DECREASE key Press NEXT LINE 34 LC NMR Accessory Installation 01 999036 00 C0800 2 7 Leak Testing the System 7 Set the pressure for the leak test The pressure may be set from 10 to 410 atm A good test pressure would be slightly higher than the expected operating pressure Typically 200 atm is used Press NEXT LINE 8 Close the prime valve if it is open Press START After the pressure has stabilized determine if a leak exists If so look for visible signs of solvent leaking at one of the fittings Note If a leak is found do not tighten fittings while the system is pressurized and do not overtighten the fitting Instead do the following Press STOP to stop the leak test b Slowly release the pressure by carefully opening the prime valve c After the pressure has dropped to zero tighten the fitting just enough to correct the leak d Repeat the leak test 9 When the leak test is finished press STOP Slowly release the pressure by carefully opening the prime valve After the pressure has dr

LC-NMR Accessory Installation

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