LCT CLASSIC A & B
Contents
1. Open Data File Queue Is Empty Spectrum Chromatogram Map Edit Samplesv File Name File Text MS File Inlet File Bottle Inject Volume Control 1 0 000 New Ctrl4n Open Ctrl 0 Save Ctrl S Oro Save As Import Worksheet Import Data amp Print Ctrl P Print Setup Exit sure aa Flow 1 000 ml min n e 8 Open an existing MassLynx project E Ej Masslynx Maggie Macarena1 w29 Inl B LCT c masslynx m fm LCT Classic training d untitled Paint F Chromatogram H2 Figure 1 Open your masslynx project 1 2 Is the sample cone clean B in figure 2 Any visible browning or other discolouration deposit close to the cone s orifice will reduce the sensitivity of C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometry Instrument Training Guide Page 3 of 18 24 10 2012 the instrument It is your responsibility to ensure the instrument and the cone is left clean such that it will not impair sensitivity for the next user Fig 2 Figure 2 ESI probe A and cone B 1 3 Choose and load a suitable IPR file for your experiment then select the corresponding ion mode Fig 3 amp 4 C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic trainin2. EN gt 5 OAR BH 13 52 Figure 9 Process mass spectra subtract smooth and centre 2 8 Spectrum Go to Tools Make Calibration choose the NaF positive ion mode reference file and then manually match peaks Figure 15 C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometry Instrument Training Guide Spectrum NaF_2 m File Edit Display Process Tools Window Help 24 10 2012 e x Page 9 of 18 s Aaleeel ral a SRO CAaR e on HSS_T3 NaF_2 11 0 188 Cm 9 44 s n TOF MS ES ai 430 9007 Make new calibration 3 55e4 Reference material 362 9189 490 9888 Reference file NaFPos ref 634 8585 C Air references 702 8475 770 8337 838 8261 906 8158 3 974 8056 1042 7804 1110 7845 1178 7570 1246 7465 1314 7452 1382 7120 1450 6951 1518 7051 1586 6923 ri 0 miz 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900 2000 Mi Dw Adob A 3M B MassL sm Googl FW Adob X EN g See HB Ws i 13 51 IA RLaAn Figure 10 Go to Tools Make Calibration choose the NaF reference file Calibration SCL4 SCL File Print Edit Display Data file NaF_2 Uncalibrated 100 Yo D Reference file NaF Pos 158 96 226 95294 94 362 93430 91 498 90566 89 634 88 702 86770 85638 84906 83 974 81 1042
3. For small molecule LC MS it is suggested to use sodium formate at a concentration of 15ug mL in 50 50 can Water with 0 1 formic acid this covers a mass range from m z 91 m z 2062 however it is more difficult to get sufficient ions to calibrate masses above m z 1500 2 Calibrating the instrument with Myoglobin or Sodium formate 2 1 To calibrate for denatured protein analysis another protein with a charge state envelope covering the expected mass range can be used BSA and myoglobin are common examples Myoglobin will provide a charge state from about m z 700 2000 Figure 7 2 2 If it is not already in operate switch into operate and switch on API A amp B Figure 5 XA MassLynx Maggie untitled File View Run Help oe Ef O EN H gt F gy Shortcut Ta Queue G Status Queue Is Empty LCT c massiyna shui te _sn shutdown ipr de Peak Display Vacuum Inlet Options View Help W amp i n al ES Source Advencafed al j Peak Display 1 Spray 2 AR E 1 0020 Capillary v fo 3000 J Sample Cone V E 220 1 Extraction Cone V l Boo H RF Lens 330 H MS Tune MS Method Inlet Method Temperatures Desolvation Temp C 201 200 Source Temp C fi 04 100 Gas Flows Nebuliser L hr Desolvation L hr 0 250 00 500 00 750 00 1000 00 1250 00 1500 00 1750 00 Ready Ready Not Scanning 0 0 Shutdown Enabled 2 KA MassLynx Maggie Macarena1 w29 Inl
4. Figure 16 Viewing UV trace through analogue channels 4 5 Choose analogue channel 1 to display the channel which collected the UV data Figure 17 C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometry Instrument Training Guide aY Document Microsoft Word File Edit View Insert Format Tools Table Window Help Type a question for help X OSE ARIERY BAP oo eA 0 o iA Noma TmesNewRomn 2 B Z U ES SSE Eee Z A e 1 Chromatogram UV test3 J File Edit Display Process Window Tools Help Z AS SoR eM OA BROAR e oS Page 15 of 18 24 10 2012 X UV Analog Chromatogram TOF MS ES TIC 3 67e5 Channell Channel 1 Cancel Add trace Align O Replace trace File O New window X s 2 2 At 1 In 1 Col 2 REC TRK EXT OVR English U S Cc nl MassLynx Jam B LCT c massly vs Experiment Set E MassLynx Help Untitled h11 I tay Documenti Mi Chromatogram Figure 17 Choose channel 1 to view UV trace 4 6 UV trace for the wavelength selected in the inlet method is added to or replaces the TIC trace in the chromatogram window Figure 18 C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometr
5. Samples lists are kept in the Sampledb subfolder in your main project folder Sample lists can be copied or renamed 3 3 The sample list has a line for each sample to be run and a large number of samples can be edited and run as a single batch 3 4 Each line requires a sample name a mass spec method an inlet method make sure all timings for the length of run match a vial position for the sample and an injection volumne note the injection volume in the sample list overrides that given in the inlet method 3 5 Editing Inlet method There are 4 important tabs circled in red in Figure 13 The first is the display status this provides information about the flow rate which pumps are running their pressures and any faults with the inlet Note the ready and OK light should both be green during normal operation A red light in either signifies a fault as for the OK light in Figure 13 C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometry Instrument Training Guide XA MassL ynx Default Default spl File view Run Help Page 11 of 18 BH 4 d D aii E Shortcut FA Queue F Status Queue Is Empty Spectrum Chromatogram Map Edit Samples v File Name File Text MS File Inlet File Bottle Inject Volume Control i DEFAULTO Default file DEFAULT Inlet Method
6. 1400 1500 1600 1700 1800 1900 2000 Saas M FW Adob EJ 9 Mi Dw Adob T 3M MassL ORN Googl fm 6 wi Ad EN g OHM Tae H 13 39 Figure 8 Direct infusion of sodium formate in positive ion mode C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometry Instrument Training Guide Page 8 of 18 24 10 2012 2 7 To calibrate 2 7 1 set cone voltage to 30V 2 7 2 Directly infuse sample using syringe pump at 5 or 10uL min 2 7 3 Acquire data over the range 100 to 2000 2 7 4 Open Spectra sum at least 10 scans then process and centre the data Process subtract smooth and centre Figure 9 u Spectrum NaF_2 e x x m Fie Edit Display MAOR Tools Window Help EA B For g Automatic Peak Detection a 5 OO Oe PFF HSS_T3 Combine 100 so Te _ Center Mass Measure 362 9189 Process All Traces Component gt MaxEnt 1 Set Adduct Mass 02 8475 MaxEnt 3 Integrate 770 8337 838 8261 906 8158 32 974 6056 1042 7804 1110 7845 1178 7570 1246 7465 1314 7452 1382 7120 1450 6951 1518 7051 1586 6923 Fa 0 mz 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900 2000 Perform background subtract on continuum spectrum FW Adob EJ 9 mi Dw Adob Esm E Massa GSM fm 4 Wi v Ad ZRLAH
7. 80 110 79 1246 76 100 D Residuals Polynomial Order 0 0 01 ppm 0 00 100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 v Fw Adob Eg 9 Mi Dw Adob Ea E O 5 mi Googl 1500 n d 1 matches of 28 tested references RMS residual 0 000000e0 ppm Miz 1600 1700 1800 1900 2000 EN P gt TMO i 13 55 gt HRLAH Figure 11 Manually match calibration file peaks 2 9 Select File save as calibration enter a filename OK choose SAVE followed by C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometry Instrument Training Guide Page 10 of 18 24 10 2012 2 10 Choosing OK on the calibration page without first saving the calibration as a different file will automatically overwrite the current calibration 3 Setting up an LC MS sequence 3 1 Arrange basic plumbing with column inline after the UV cell if using UV and mass spec detection Figure 12 Column selector valve Autosampler T il Mass spectrometer FOUN detector i E Fraction colector Flow splitter HAPLO pumps Figure 12 an appropriate diagram for plumbing of HPLC 3 2 The sample list is loaded on the main masslynx page A new sample list can be opened or a new one started using File open or File new respectively
8. Data Format Continuum X Scan per 16960 p Close Calibration 250 00 500 00 750 00 1000 00 1250 00 1500 00 1750 00 2000 00 2250 00 q Press for Standby C Ready Vacuum OK Operate h T ZOTI VAIQUe ZU Vdldieria 009 39 H2021 Maggie200to Macarenal Ze 25 000 40 11162008 Ph Maagie200to Macarenal 23 25 000 w Ready Not Scanning 0 0 Shutdown Enabled 2 E O LCT Classic training d untitled Paint bd MassLynx Maggie Untitled w29 Inlet M LCT c masslynx sh Figure 6 Acquire data manually 2 5 In order to check the mass for Myoglobin de convolute the charge state envelope First process smooth subtract and centre using the process tab Then select process components and manual fill data in Alternatively the Maxent 1 tool can be used to de convolute the spectra Note this works by taking the spectra actually displayed in the mass spectrum window and not all the date saved in the data file Relevant reference values for the ions in the sodium formate mass spectrum can be found in the masslynx folder opened with notepad from C MassLynx Ref note these are the same values referenced when making a calibration 2 6 The chromatograms in Fig Figure 7 amp Figure 8 show examples of the spectra you should see for Myoglobin and Sodium formate C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes F
9. Edit Shutdown or Startup Shutdown QuanLynx BioLynx Tools Instrument Startup Q DEFAULT 1 10 000 Default h11 Inlet Mothod 24 10 2012 File View Tog LC Help Dw EMATEN Status Inlet Autosampler Detector Display Status information Indicators Running Pump On Inject Cycle Ready OK LC Run Time mins ea Flow Rate rl min Pressure bar counter ec i Column Right C fl Sample Temp C 0 000 0 000 0 0 r Gradient A 100 0 m PDA Detector Mode Not Installed Scan 0 Figure 13 There are 4 important tabs circled in red v Only Error Shutdown Enabled E EN Fe BB A 14 10 3 6 Editing MS method Right click and select edit on the MS File group Figure 14 This will open the MS method which can then be modified alternatively click on MS method in the Instrument section of the Main Masslynx page Set the mass range for data collection and length of time mass spec data should be collected for Note this does not have to be the same length as the LC method run time C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometry Instrument Training Guide Page 12 of 18 24 10 2012 KA MassLynx James McCullagh McCulla
10. Mass Spectrometry Instrument Training Guide Page of 18 24 10 2012 CRL MASS SPECTROMETRY FACILITY USER MANUAL LCT CLASSIC A amp B 1 Floor Mass Spec Lab 00 097 This is a guide to using the LCT classic for those who have received training If you have any questions or problems whilst using this instrument please contact a member of the CRL Mass Spectrometry Facility Staff C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometry Instrument Training Guide Page 2 of 18 24 10 2012 CONTENTS 1 Checking the system is working properly Page 2 2 Calibrating the instrument using Myoglobin or Sodium formate Page 5 3 Setting up an LC MS sequence Page 10 4 Using UV and MS detectors in sequence Page 12 6 Cleaning the sampling cone Page 16 9 Contacting MS staff Page 18 1 Check the system is working properly Make sure the Masslynx program is running and in operate with the appropriate IPR file masslynx method loaded for positive or negative ion mode Note when you have finished using the instrument it should be left with the masslynx program running and in operate 1 1 Open your masslynx project File Open project select your project name Fig 1 A warning message will appear click yes ___ View Run Help E La s s y a UD a soat B oue Project Wizard amp Status
11. eb2010_241012 doc Mass Spectrometry Instrument Training Guide Page 7 of 18 24 10 2012 Spectrum 11162010_myocalibration l 7X E Fie Edit Display Process Tools Window Help s Boal eeelral a SRO CAOAR Ea 11162010_myocalibration 1410 14 108 AM Cen 4 80 00 Ht 5000 0 0 00 1 00 Sm SG 2x2 00 Sb 12 35 00 Cm 1344 1521 TOF MS ES 400 847 78 1 17e4 807 45 392 34 941 89 110 77 W Manual Find Components Find Component Found component Peak1m z Window Threshold Molecular Mass 997 24 41 99 1 00 Da 1 00 BPI Peak 2 m z Reiect Missing peaks Delete All 737 31 997 24 CI Dimers 2 00 sd s 1059 53 706 63 4130 10 amp 1210 77 1303 82 678 39 1412 37 1540 62 1694 47 1882 42 2116 87 0 mz 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900 2000 2100 aaa E O fa LCT Classic tr untitled Paint r MassLynx M Untited w29 LCT c massl T Chromatogra MassStore on J 11 16AM Figure 7 Denatured Myoglobin spectra Spectrum NaF_2 File Edit Display Process Tools Window Help sBoaleaeelral a BROAR eN 8 x HSS_T3 NaF_2 11 0 188 Cm 9 44 TOF MS ES 430 9007 3 554 100 362 9189 498 8888 634 8585 702 8475 770 8337 838 8261 906 8158 z 974 8058 1042 7804 1110 7845 1178 7570 1246 7465 1314 7452 1382 7120 1450 6951 1518 7051 1586 6923 Fa 0 mz 300 400 500 600 700 800 300 1000 1100 1200 1300
12. f LCT Classic training d LCT c masslynx sh 5 untitled Paint Figure 5 Switch into operate A check gas on B 2 3 Directly infuse the appropriate standard solution at about 0 01 0 1mg mL or 10uM using the syringe pump and a 250uL syringe 2 4 Acquire data manually for directly infused sample Figure 6 C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometry Instrument Training Guide Page 6 of 18 24 10 2012 XA MassLynx Maggie maggie13082008 SPL File View Run Help 2 OBH amp gt OOD shota aN Queue F status LCT c masslynx shutdown shutdown ipr BAA File IonMode Peak Display Vacuum Inlet Options View Help 8 D h Sim oana s Veen I z ES Source Advanced Engineer Seles E i m Source v Capillary jo 2000 v Sample Cone 24 22 a Tune Page Acquisition z Extraction Cone V jo 3 H v RF Lens 230 H Data File Name type a file name here S z Browse Ww i Temperatures Text additional text herel Desolvation Temp C 210 200 Origin s Source Temp C eg fioo as 7 m Mass Start Mass eo Gas Flows End Mass 2000 Nebuliser L hr Desolyation L hr J m Time E Duration mins 35 E Scan Time s f CE 103 683 Inter Scan Delay s 0 1 m Data
13. g documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometry Instrument Training Guide Page 4 of 18 24 10 2012 XA MassLynx Maggie untitled File View Run Help gt DHZ D I Z shota Gy queue S status Queue Is Empty LCT c masslynx shutdown shutdown ipr File IomMode PeakDisplay Vacuum Inlet Options view Dae g Oly al ES Source Advanced Engineer Peak Display 1 Spray 2 Inlet Method Source Capillary v fo j2000 T Sample Cone ja z2 Jm Extraction Cone 3 a H RF Lens 330 H MS Tune MS Method m Temperatures Desolvation Temp C 208 200 Source Temp C fing fioo Look in amp ACQUDB v 6 c Fae E default ipr Positive ion default ipr HT_tune page pos ipr m Gas Flows jmc_1 ipr Nebuliser L hr Desolvation L hr Negative ion default ipr NEWMCPNEG ipr NEWMCPPOS ipr Z 4 File name Positive ion default ipr Files of type Jinstrument Parameter Files ipr Cancel 0 250 00 500 00 750 00 1000 00 1250 00 1500 00 1750 00 2000 00 2250 00 Press for Standby E Ready Vacuum OK Ready Not Scanning 0 0 Shutdown Enabled E O KA MassLynx Maggie Macarena 1 w29 Inl m LCT Classic training d LCT c masslynx sh 5 untitled Paint Figure 3 Select a suitable IPR tuning file for your experiment from File open ma
14. gh _PeptideMapping_Feb2010 SPL File view Run Help Zz F x H amp gt LJ Ii Shortcut ra Queue gt Status Queue Is Empty e MS Status Spectrum Chromatogram Map DriftScope Edity Samples v 5 File Name File Text MS File Inlet File Bottle Inject Yolume Control a 1 NaF_test ooplnjection_2mins Loonlniection 21 5 000 Browse i oom 3 0 000 0 00 E 4 Cut 0 000 Eo ea 5 Copy 0 000 a Paste 6 0 000 2 x Ready 7 Add 0 000 gt 8 Insert 0 000 E 3 Clear Selected 0 000 Z wn 10 Customize Display 0 000 Time Pressure 11 0 000 0 00 min 5284 psi p 12 AutoSampler Bed Layout 0 000 a 13 0 000 14 0 000 Flow 15 0 000 0 200 ml min 16 0 000 7 0 000 18 0 000 100 0 0 0 13 0 000 20 0 000 21 0 000 22 0 000 23 0 000 24 0 000 25 0 000 26 0 000 27 0 000 28 0 000 29 0 000 30 0 000 31 0 000 32 0 000 33 0 000 34 0 000 35 0 000 36 0 000 a nanan Mi Ready Not Scanning 0 0 Shutdown Disabled 2 4 MassLynx James M Crim McCullagh_Peptides6 ons Sa Experiment Setup c E c ha J SYNAPT HOMS McCullagh_Peptides gt ACQUITY UPLC C E t Set ALGEN Figure 14 Editing MS method from sample list 3 7 Editing and saving sample lists The sample list can be saved and opened using File open 3 8 labelling bottle vial number in sa
15. ke sure your project is selected YA MassLynx Maggie untitled File View Run Help a ba L EN H BD gt g gy Shortcut Fa Queue status Queue Is Empty LCT c masslynx shutdown shutdown ipr File POUNCE Peak Display Vacuum Inlet Options View Help ji Sloe nced Engineer Peak Display 1 Spray 2 Inlet Method Source Capillary V fo j2000 IE Sample Cone f js 220 1 Extraction Cone pa Bo H RF Lens 330 H MS Tune MS Method m Temperatures Desolvation Temp C 203 200 Source Temp C fi 04 fi 00 Gas Flows Nebuliser Lhr Desolvation L hr 0 250 00 500 00 750 00 1000 00 1250 00 1500 00 1750 00 2000 00 2250 00 Press for Standby T Vacuum OK 4 Ready Not Scanning 0 0 Shutdown Enabled 2 E O Ki MassLynx Maggie Macarena 1 w29 Inl LCT Classic training d LCT c masslynx sh 5 untitled Paint Figure 4 Choose the correct ion mode for your experiments lon mode ES or ES C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometry Instrument Training Guide Page 5 of 18 24 10 2012 1 4 Before setting up a sequence of injections you should directly infuse a standard sample to check the mass spec is functioning normally and to calibrate if necessary
16. lassic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometry Instrument Training Guide Page 18 of 18 24 10 2012 Dry the cone and nozzle using mtrogen To minimise down time fit a spare sample cone obtainable from Micromass at this stage If material has built up on the exhaust liner and cleanable baffle Remove the cleanable baffle and the exhaust limer A Canton Do not attempt to remove the baffle without first removing the sample cone Clean these components or obtam replacements Fit the cleaned or the replacement exhaust lier and cleanable baffle to the ion block Refiting the sample cone is a reversal of the removal procedure 6 Contacting MS staff For instrument related questions or if you have a problem with using the LCT contact one of the mass spectrometry research technicians James Wickens Colin Sparrow or Lingzhi Gong in the basement mass spec lab on 75710 Alternatively or for any other enquiries please contact Dr James McCullagh Head of the Mass Spec Lab on 75657 C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc
17. mple list Make sure the sample list contains an injection volume and a vial position in the autosampler 3 9 Start LC MS run Click on start on the MassLynx main page make sure Files names are not repeated in the sample list and spaces dissallowed characters are not present avoid dots or commas use underscore or dashes instead 4 Using UV and MS detectors in sequence 4 1 To add the UV detector open the inlet file in the sequence right click and open C Documents and Settings ms staf My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometry Instrument Training Guide Page 13 of 18 24 10 2012 kd MassLynx James McCullagh jmc_test SPL Giles File View Run Help lIBH 4D Bill shortut By queve B Status Queue Is Empty Instrument Spectrum Chromatogram Map Edit Samplesv E File Name File Text MS File Inlet File Battle Inject Volume Control a E g 2min_test jmc_2mininlet 1 25 000 A 1 000 5 Inlet Method e amp File View Tools LC Help c Tr Alo 7 T gt MS Method DSHS Gas 2 O MS Tune j GP M UY Detector i Analog Edit Shutdown or Startu n i Wavleneath nm Zero Offset 5 W Response ms e0 Attenuation m4U 1000 i Autosampler Startup 4 E Detector E For Help press F1 Options tE To aooo 0 000 0 000 nom J an
18. n 0 000 0 000 ann ann 36 0 000 ann 0 000 40 0 000 Mi Ready Not Scanning 0 0 Only Error Shutdown Enabled E O ha MassLynx James Mc LCT c masslynx ja ts Experiment Setup c B MassLynx Help Untitled h11 Inlet M Figure 15 Editing UV detector parameters 4 2 Select the edit detector parameters tab see Figure 15 4 3 Choose the wavelength required and save the inlet method 4 4 To view the UV trace open the chromatogram window select Display from the menu bar and then Analogue Figure 16 C Documents and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Page 14 of 18 24 10 2012 Mass Spectrometry Instrument Training Guide amp Document1 Microsoft Word 2 File Edit View Insert Format Tools Table Window Help Type a question for help X OSE BR SRF TERY noo Ano gt 2 A Noma O gt TmesnewRoman v 12 BZ uE EESE e Chroraatogram UV test3 m E Fiy Eat RER Process _ Winde aye W ABRO CAVW nme UV te TOF MS ES TIC 3 67e5 Customize Toolbar v Toolbar v Status bar Move To Last Move To First 8 00 9 00 10 00 11 00 12 00 13 00 14 00 B 4 Page 1 Sec 1 1 4 At Ln Col REC TRK EXT OVR English U S gt LCT c massly va Experiment Set E MassLynx Help Untitled h11 I tay Documenti Mi Chromatogram
19. ocuments and Settings ms staff My Documents Chemistry Instruments LCT classic and premier LCT Classic training documents LCT CLASSIC training notes Feb2010_241012 doc Mass Spectrometry Instrument Training Guide Page 17 of 18 24 10 2012 Removing and Cleaning the Sample cone Caution The sample cone ts a delicate and expensive handled with extreme care It is not necessary to vent the instrument to remove the sample cone The source block incorporates an isolation valve for this purpose To remove the sample cone proceed as follows Follow the procedure m the previous section to gam access to the sample come Using a suitable flat blade screwdriver rotate the valve by 90 into its fully A small improvement in the analyser vacuum may be observed as a result of this operdanon The isolation valve is in the closed position when the slot is perpendicular to the direction of flow Disconnect the cone gas mlet hme if fitted Take the sample cone extraction tool supplied m the source spares kit and screw it to the flange of the sample cone Remove the two sample cone retaining screws using a 1 5mm Allen key and withdraw the sample cone and cone gas if fitted from the 10n block Remove the extraction tool and separate the sample cone from the cone gas nozzle Place both components m an ultrasonic contamme 30 30 acetomitnle water or 50 50 methanol water C Documents and Settings ms staff My Documents Chemistry Instruments LCT c
20. y Instrument Training Guide Page 16 of 18 a Document1 Microsoft Word 24 10 2012 gt File Edit View Insert Format Tools Table Window Help T p Be Osean 4RY Ses ho o BWMB 10 B A Normal TmesNewRoman 12 B Z U 3B t2 2 E Ht O Z AY e Chromatogram UV test3 fi File Edit Display Process Window Tools Help SIAR SR e rAWI OA EROA we OB UV test3 Channel 1 An1 5 68e5 15 00 UV test3 TOF MS ES 11 00 v t Qo 3 3 At 1 Ln 1 Col 2 REC TRK EXT OVR EngishU S GJ Start E O KA MassLynx Jam LCT c massly vs Experiment Set E MassLynx Help Untitled h11 I tag Documenti Mi 4 Chromatogram ad 4 m Figure 18 UV trace for the wavelength selected in the inlet method is added to or replaces the TIC trace in the chromatogram window 5 Cleaning the sampling cone 5 1 The sample cone should be cleaned regularly whenever a discolouration can be seen around the orifice this may be every 24 hours for continuous use or more frequently if protein samples or buffers are run at higher concentrations 5 2 Before removing cone make sure isolation valve is closed 5 3 The cone should be removed and each metal part sonicated for 20mmins in 50 50 MeOH with acetonitrile and 0 1 formic acid There is a sonic bath in the basmement mass spec fume hood Blow all parts dry with air before re assembly 5 4 After re assembly make sure isolation valve is open C D
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