UPLC/MS/MS システム取扱説明書 分析操作手順 （定性 編）
Contents
1. MS LOAD INJECT 1 Inlet method ACQUITY Additional Status 6 HP wyhp HP w2200 HP wpda Inlet Method 3 xl File View Tools LC Acquity 5ampler Help Status ACOUITY additional Status ACOUITY Binary Solyent Manager ALUUITY Sample Manager ow 2 ACQUITY Binary Solvent Manager ra me sesamer a unication jj Shutter 2 NN 3 2 Start up system Far Help press F1 System Startup Prime Solyents Equilibrate to Methad Optional Characterize Valume A B Solvents 4 System Startup Prime A1 Prime B1 sealwash I Prime a2 Prime B2 J Strong wash cycles SO Prime Solvents a 1 Duration of prime I Weak wash I Sample syringe A B Solv2. Project ACQUDB 23 znnngn3 w2znn 20070903 wpda Alicia da Defa gyHgH3 wwhp licin_nnstrun5Hrmnin wwhp hie al mlicin w22nn anthncyanin_inlet wzznn ESCI allicin wpda anthocyanin_ inlet mpda E5C1 licin wwhn anthnrwanin_inlet wyhn E5C1 File anne soukenwyhp Save as uhe Methad Files whp maz wpda Lanicl SW save jm ACGUDB hesktnp Emin w22nn hefai 9 a FOrmin pda hie al 3 Flappw 5Imin wyhn hieFal En uatem iF let 22hn ESC1 let wpda ESCI lat wwhp ESCI 8 LE LL Fe Te Shared Documents kp Dncumenls lu Nehwnrk Places aCe Gawe ast 23 15 3 MS Method MassLynx Status 24 MS Method Eile View Run Help pi J dl shortcut Queue GB status Queue Ts Empty Spectrum Chromatogram Map Edt Samplesr File Name File Text kMS File Inlet File Eottle Inject Volume lt 1 II M Tuna MS Method Inlat Method 24 MSMethod Filel UPL
3. 5 4 Waters IC PC UPLC 1mL
4. M8 je 7 147 37 0 B458 519527 B096 44 81 28 40 659 20 7510 05 794 74 100 200 3HI 4nH S00 5 700 gD Process Combine Spectruml Average MS 68 1004 ng1 11 43 11 84 Time i 67 MS ram Rosemary solid new 40ugml 1 x Fle Edk Display Process Window Tools Help Sa a megate Purity Rosemary_solidii smoothi Subtract Apne sm 13 Nov 2007 14 07 59 Rosemary solid ne Process All Traces 1 Scan ES NE 9 88 i ww lt amp 2 3 Mi 2 2 5BeB MS gt eg Average J 10 04 11 43 11 84 0n33 110 215 374389 4 519527 B096 44 8328 4 RN nbtract 4 WINE SN 268 2 803 12 13 1 00 2 00 3 00 4 00 5 00 6 00 7 HH 8 nH 9 0H 10 00 11 00 Time oe
5. A1 A2 B1 B2 souken wy hp souken w2200 souken wpda Inlet Method autosampler Detector 5 psi 2 Sample ME Column Q 000 mlimin BZ 0 35 Off autosampler Detector 27 Inlet method 17 ACQUITY Additional1 Status Console Delta 40 Inlet 2 Console View LC PT PR Help ALUUITY Binary Solyent Manaqer 8 ee ACOUITY Sample Manager ow ES 2 B00 Sample OE Column 0 200 mLjmin 50 0 95 5 0 4 rh NNS mn oo GOmaole rr UPLC Lee System Sample Manager Column PDA Detector Ploks Maintenance Counters Logs Binary Solvent Managerl Binary 5olyent Manager Flow 70 0 0 0 0 0 mL nnin Stop Flow Sample Manager PDA Detector MA Off wi TE Off Column Shutter Closed Syst
6. 4 Method Cone Voltage Cone MS 5 MRM MSMS 9 7 V Chromatograml 63 File Wiew Run Help Open Project Project Wizard Wa MassLynx souken souken3 5PL I pL Ul 2 Shortcut 3 Queue F status Open Data File Mew Ctrl N open Ctrl O Save Ctr 5 Save As Sample List Properties mpty pectrum Chromatogram Map Edy Samples v le Inlet File 1 est D80214 2 ltesH Bottle 2 F 8 Control lt Inject Valume 5 000 souken3 souken Import worksheet filel IOpen Import Data SO Print Print Setup CtrlTP Exit Rn Function MS Method Chromatogram Data Browser Flg Mame D
7. Hile Name Filel Save as UV MS TITC 94 Chromatograml Diode Array TIC Display TIC 58 Chromai czram stU31 Clme sm 2 Dinde Array Range 7 57 4 12 7B_14 45 15 00 58 Diode Array TTIC MS 59 Spectrum Pest03 File Edit Display Frocess Tools Window Help X 0 A CC IPesticide Mix PEStH3 458 B BH51 1 Scan ES
8. m combine spectrum rE a a File Fiosemarnlsc Furnctior 1 MS i verage Farncel a Peak ni Mulliple Awerage sBaralin ubtract W TH Subtract 2 68 Combine Spectrum 44 Spectrum Rosemarytsolid tnew tt 40ugmh _ File Edit Display Froress Tnnls Windmw Help LA RoseImary_solid_new_d0ugJml YAB915 13 how 2007 14 07 59 RnsEITmaPTS ldHiFTEWTHHu FI BE TH BT Im TB5HI T1 1 1 ca ES 1nn 31 1 d dB 112 5d 0 421 15 315 91 rc rr Nc 399 12 RnsEITmaPTS liHTTTEWTTHHu FI BEG 1 1nn 11 1 1 Scahn ES HBEb 69 45 Ifilel IPrintl test 080214 345 12 806 Cm 344 348 2 Scan ES 331 29 1 95e7 100 332 44 333 47 399 34 469 41 663 45 824 07 961 39 688 64_752 97 998 11 0 test 080214 346 12 834 Cm 344 348 1 Scan ES 328 22 7 58e7 287 14 Se 440 24 355 11 695 44 368 35 442 26 so1 20 41 47 0 9 test 0802
9. 7 TE TO 48 SNEOCD NCC AGGGc Gl 49 4 Xbridge 1 LC MS MS Ultra Performance LC ACUITY UPLC Waters Micromass Quattro Premier Mass Lynx4 0 UPLC MS MS 1 UPLC 1 2 LC MS 2 3 LC MS MS 3 UPLC 2 MS 1 UPLC MS1 EEE LC MS LC MS MS UPLC gt MS a 2 LC MS Collision MS1 Cell MS2 UPLC MS MS 3 LC MS MS 1 UPLC LC MS
10. SW 90 10 100 2 100 mL 1 Ns UPLC Al A2 B1 B2 Inlet Method UPLC Tnlet method MassLynx Status 4 Inlet Method Inlet Method WAMassLynx initial untitled Jlx Fle View Run Help 2 shortcut Queue status Queue Ts Empty MSSftatuS Spectrum Chramatagram Map Editr Samples FeNamwe File Text MS File Inlet File Bottle Iniect Volume 5 EE 0000
11. Dwell 0 2 dmm Cone IAddl 1 UCS CV UseTune Cone 5ehtins Fetentior indo hdirns Start s En Rl RinEE ee FEIBEIIT g Bi ClearAl Frobe Temp in Method Retention Window APCI Probe HE arr 52 SIR 6 Hilel ISave Asl 53 Enp TinnEnt setup FitDFnjECELTnaEsuFuii hrniar uh uunlrnnly Enh Dptions Tnnlhars Functions Help et X a MS can SIR MR Daughters Parernts heutral Loss Meutral aI LUrwe Tatal Run Time 15 0n H 10mins Ng Type IrFnrmaFinr SScan Time 0 00 to 15 00 Mass 100 00 to 2 nn ES d IS Scan Time O 00 to 15 00 hass TO0 00 to 581 HH ES I 5 SIRofmass195D0 TimeDOOto sODES 5 SRnfmass 195 00 Time 5 00 to 10 00 ES ES Scan Time OD 00 to 15 00 hass 100 00 to TOOD DD ES ES Scan Time OD 00 to 15 00 ass TO00 00 to 2000D DOD ES 53 MS Method 9 MS 1 UPLC 0 2 0 4 ml min
12. 6 7 1 Inlet Method UPLC MS TUNE MS LOAD 2 Operate MS 1 MassLynx Status 43 MS Tune Queue Ts Empty Spectrum Chromatogram Map Edit 5Samhplas File Nam mi File Text MS File Inlet File Bottle Iniect Volume MS Tune MS Method Inlet Method 43 MS Tune 26 2 MS tune APIGAS Ns 44 COL GAS Collision Gas Flow API GAS COL GAS Quattro Prennier CDI initi acqudb mrm ipr PP LR 1 Daughter Scan wis5e 112 2 5
13. Acouity PDA Detector Gearal zp hanne nalng Out Ewents 1 Run Time BB A Rahng PFgsnlutinr Enable 3D Data ey Gampling Rate Filter Time Corstant Exposure Time PDA UV 1 PP NN eee Ie lm Use UW blocking filter thelow 210mm 190 500 a nm MS CR EC 20 Detector 3 2 D Cannels Modify ACQUITY PDA Imstrument Method Acouity PDA Detector CE 2D Channels Analog Out Events Channel 1 12 gt nm resolutic 21 Channel 2 PDA 21 2DChannels Analog Out Events 4 OK p8 11 14 Inlet Auto sampler Detector
14. ESI EST APCI APC1 LC MS Cone Capillary STR LC MS MS Colhsion energy MRM IT UPLC 1 Inlet Method MS Method Oo Ol 6 MN UPLC MS MS UPLC MS MS MS LOAD A1 A2 A B1 B2 B SNW WNW
15. OK amples lumber of Samples lt RN UG ah BE hEF HLT er 3 IDEFAULT03 4 IDEFAULT04 5 DEFAULTOS 6 IDEFAULTO5 RS MMU 2 33 Auto Sampler Bed Layout utosampler Bed Layont Plate Bed a 4am Holder a am Holder Samples gt Addl 34 Oueue Is Empty Spectrum Chromatogram Map Editr Samples yr File Name File Text MS File dd 1 Default file DEFAULT Insert Delete Fill Clear Calumn k Fnrimat by Fiancel Default ig Default ilg helault ilg helault ig helault le helault lg Number nf Samples 34 Samples Sah DEFALILT DEFALILT DEFALILT DEFALILT DEFALILT DEFALILT DEFALLT DEFALLT hEF HLT IEF HLT IEF HLT IEF HLT 1 1 2 1 3 1 4 1a E 35 21 1HLHHI 1HLHHI 1HLHHI 1HLHHI 1HLHHI 1HLHHI Number nf Injectinns autosSampler Bed Laynut Ln Jume I0 0 Filel gt Save As 86 A Masslynx souken souken3 SPL UH i 1 Fle View Run Help W Open Project PP MN 2 5hortcut Queue GLaus Project Wizard a Queue Is Empty ae nectrum Chramatagram Map E
16. RnsemaryiL DAD Chromatogram IDescriptionl OK Dab Chromatogram x Flg Lescrinhinh nm FnsemarlLinuldl HF Cancel Filg Funicligft 5 Diode Ara 235 400 ah hrnmatnnram Indo in Parls per mllinn bs tid nm HH trace Helacg trace Newi windaw Tic analaa Remoye Real Time Update ChraTaal DDaToal Range v Pointer View Peak Annotation Customize Toolbar Toolbar wv Status bar ave To Last Faye To First Traces T OD 00 1 1p 33 gn 2 00 Rnsermary Liquid nWw 2ugml 3 4 50 4 84 5937 3B BBH 2 00 Select mass wavelength chromatogram 4 00 Chromatogram Rosemary Liquid new 200uqml I x File Edit Display Process Window Tools Help x 0 A One sR ml 13 Nov 2007 13 35 58 5 DindB Array Range 4 561 9 75 13 38 8 95 4 00 5 00 8 HH 10 00 12 00 14 00 1 Scan ES 9 55 TIC 2 82e8 9 07 10 12 13 24 gt 13 54 13 77 14 25 12 15 5 43 11 76 Time 5 00 HH 10 00 12 00 14 00 65 UV 41 filel Printl test 080214 5 Diode Array 11 14 350 7 0e 2 Ra
17. Scan Time 2000 Da Scan Time 0 5 31 MS 1 20 30V ES OK 2 MS Scanl ES ES OK 3 2 AP AP OK RSI APCI 1 10 20 30 40 50 60 V STR PETETT III NN Lhannel Malhnd WW SIR SIR Mass Diwel Cone onizatior Mag Es lt mal EE ai Intmr hannel hialan nm A 0 a 95 p 2n Inter 5can hglaw ld5 Mass a
18. 1 12 Inlet 4 Time min A B Curve HPLC UPLC ACQUITY UPLC Columns Calculator 18 TT LIFELC Colg 13 ACUITY UPLC Columns Calculator ACQUITY UPLC Columns Calculator Gradient Separationl HPLC Column Length Column Diameter Particle Size MW of Sample Flow rate Temperature Injection Volume Delay 6 Time min B Calculatel THE 5CIENCE DF WHAT POSSIBLE E gt About thi lculat Select a calculatinn Ua St References gt Isocratic Separation Brochures amp Literature Gradient Separation gt Wisit Wat
19. LA ER CDO CE ESI CG ESI ESI ESI ESI APCI 48 3 0 1 MeOH MeCN 0 2 MeOH MeCN 0 05 TFA IMeCN 10 mM MeOH MeOH MeCN 0 5 PDA 88 4 Xbridge 49
20. 5 Bottle 6 Injection Volume a Masslynx DEFAULT Default_spl File View Run Help 8 Shortcut Queue sews Queue Ts Empty Spectrum Chromatogram Map Edit Sampl amples v File Name FleText MS File Inlet File Bottle Iniect yal 1 DEFAULTOT Defaulfle DEFAULT DEFAULT Inlet Method Flow n nnn m rnin 100 0 wv Only Error Shutdown Enabled 831 Bottle Bottle Auto Sampler Bed Layout 32 32 AutoSampler Bed Layout 20 1 or 2 9 1 1 2 2 2 AO 1 A 1 0 0 IReplacel Bottle 2 A1 1 2
21. L DEFA amp AULTOT Default file DEFAULT DEFAULT 1a 1 10 000 2 DEFAULTO2 Default file DEFAULT DEFAULT 1 4 2 10 000 DEFAULTO3 Default file DEFAULT DEFAULT 1A amp 3 10 000 DEFAULT04 Defaul file DEFAULT DEFAULT 1 A 4 10 000 5 DEFAULTU5 Default file DEFAULT DEFAULT 10 000 DEFAULTUE Default file DEFAULT DEFAULT 1 5 10 000 DEF amp ULTO Default file DEFAULT DEFAULT 1A 7 10 000 40 Chromatograml J a MassLynx Matsufuji untitled Fle Miew Run Help Sl 0 2 so oee Fs 41 mW Operate A ne sR Inlet Method Chromatogram Pest03 File Edit Display Process Window Tools Help PestH3 2 DindE Array 3 24 Rande 7 57 4 33 8 3n BB 3 AU 5 32 10 70 5 98 11 69 1 93 12 75 10 14 46 5 00 10 00 15 00 20 00 25 00 41 24 IChromatograml UV TIC IDisplayl ITTC TIC Chromatogram 42 TIC Chromatogram IDiode Array
22. Inlet method Is LC Reset Communication 10 Detector HP wyhp HP w2200 HP wpda Inlet Method Status Indicator i Pump on Ready OK autosampler Detector 10 PDA 2 Inlet Method Inlet Method Inlet Auto sampler Detector 11 souken wrhp snuken w soukenwpda Tne ethnd 11 Inlet Method 8 Inlet 12 1 General Solvents i a nlyents Fressur Limits Fe lo gm I cetonitrile Highi 5nnn Fadient odin UT Hinary Solrent Hanaeerinstrument Hethod Binary Solvent Manager Run Time 1 00 mi 2 IGradient Flow MS 0 2 0 4 mlmin cms 3 2 anc El
23. OK 42 UV Add trace OK 2 2 2 ic 1mgjiml_te analaa Remowe Rose test Iv Real Time Update ChraTaal DDATnnl 2 Dinde Array Rangde 2 89BB 2 Range wv Pointer Wiew DDe Peakannokatian Customize Taalhar Toolbar m 1 5 wv Status bar Mowvwe To Last Move To First 1 UE IS TIL Lhrnnnatnnrann races Fllg Hose Est Funclinn HK 1 5can OD 581 ES Laricl la WelEUIPIIIEI Filg dd tace BPI Ehramatnaram Hehlacs trace lew windnw 42 TIC Chromatogram UV 25 LC MS MS 1 2 1Inlet Method 3 MS TUNE 4 MS Method 5 MS
24. HP wyhp HP w2200 HP wpda Inlet Miethod File View Tools LC Acquity 5ampler Hel OO ok OO Ready A Time Pressure Status acauiry Additional Status 0 psi MS Tune MS Method Inlet Method im O 00 min Indicators Pumps a Running Time mins Flaw a 0 n0n ml rnin Pump Hn A Iniect Cycle 0 Flow ml min 100 0 0 0 Feady Autosampler nK Pressure psi Detector Scar Mode NotlInstalled Detector For Help press F1 4 Inlet method PDA 5 1 2 3
25. Inlet Method IFilel ISave asl 22 ja MassLynx souken souken3 5PL I File Miew Run Help Ea gt 1 dl shortcut Queue F Status Queue Ts Empty ss MSStatus Spectrum Chramatngrarm Map Editr Samplesv File Name File Text MS File Inlet File Bottle Inject Valume Control lt 1 estIGH2142 ltest1 souken3 souken 2F B 5 0U0 EE Operate In nn LCStatus souken wyhp souken w2200 souken wpda Inlet Method OK Ready File iew Tools LC Acquity Sampler Help New CtrTNN Open Ctrl O Save Ctrl 5 in Time Pressure 0 00 min 5 Psi Print Ctrl P Print Preyiew Print Setup Flow 1 Ci Project souken w2200 souken wpda 0O 000 ml rnin 2 CProjecty souken w2200 1souken wpda 3 CiProjecty Msouken w22 H souken wpda 4 CiIPrajectl Rosemary_30ta8nMeOH wpda Properties 7 H 30 H Exit Fressure ps OK Scar Made ldle Detector Detector Save he method with a new name Ready Not Scanning oO D Only Errar Shutdown Enabled 2 22 Inlet Method
26. Desalyalion Temp C 4 E 4UH Fas Flow rc VaB915_M52HMRES i MMS jpr waB915 ACCNMASS ipr VAB915_MS25ETUP j inFusion_ 51 ipr VaB915_ESCI ipr VAB915 NEGSENS ipr inFusion_M5MS ipr VaB915_IDLE ipr WAB915 POSSENS ipr RM ipr vaB915 MS1HMRES ipr waB915 515ETUP ipr Desalyatian L hr Cone L hr Syringe M 1 File name leipnp Fump Flow uL min 5 Syringe Status Files of type instrument Parameter Fies py A Cancel 60 MSTUNE MS Method EneErirnEnt Setup cprojech matsufuji pro acgudb upleconlyErnp File 61 MS Scan SIR MS MS Edit Qtinns Tnnlhars Funrtinns Help X EScan a MM paughters Farents heutralLass heutralGan gt Tntal Run Time 5 HI 1 1Hmins hs Scan Time HHH tn 15 00 hass 1IH nLI to S81 00 ES 61 MSMS MS Method 36 NIS SMG x Method Daughter Scan MS File 62 ass mz Daughters of Innlgahinn hode E5 Data centoid n
27. lsn 55 2 99 hs ho 4 Extractor v M RF Lens MM bi 98 Temperalures Saurce Temp h 2D Desalyalion Temp C ss nn FEas Flaw Desalyalian IL hr Cone L hr ll LL 8UH gn 50 48 Syringe Pump Flow luL mn 15 syringe Status 100 0 105 0 110 0 115 0 120 0 H cquire Fress for Standby Ready lyacuum ok Operate 47 MS Operate N API GAS Ns Source Temperature MS 29 IMS Method MS MS Scan SIR MS MS 1 Mass Lynx MS Method 48 ER initial untitled x Fle View Run Help a 3 2 1 D gt 2 Shorteut Queue Status Queue 1s Empty ss MSStatus Spectrum Chromatogram Man Editr 5Samnles File Name File Text MSFle InletFie Bottle Iniect Volume
28. Time HH to 15 00 Mass TO0 00 to 2000 00 E hiS Scan Time 0 00 to 15 00 Wass 100 00 to 2000 00 ES MS Sa Time HHH to 15 00 Mass TD OD to S81 00 ES SIR nf mass 195 00 Time tn 5 00 ES SIR nf mass 195 00 Time 5 00 tn 10 00 ES 50 MSMethod 5 MS Scan x Mgthnn 51 MS Innigalinri nde Data centroid cah Diuration secs ran Time nt Inter 5can Delay ns Pcl Frobe EEE FnE Gettings Frobe Ternp an Eee Mass m z 3000 Time Mins UPLC Run Time gm Tuns Pang Lnne nltang 3n se Enng nltane Ramp HI Cone Voltage 2 10 50 V 51 MS Method ES ES AP AP APCI MS ESGi mode Scan Duration secs 1 5000 Da
29. cah Duration secs 1 Mass m z MS Start End Sah Time nter Scan Delan Collision Ehergn UseTune Page Lnllsinn E hergy iw UseCollision Energy Ramp Ea aFcl Frnhe Ln nltane Llsg Tune Pang Lnre nltang Lisa Lane yulkane Hamp 2 Time Mins UV MS 0 1 62 Daughter Scan ee Pa ellis Frnhe T emp II MSMS 3 Collision Energy 10 20 30 40 50 60V
30. 1 00m Derate 0 0 n m OO ok Ready m s O nN Time Pressure 0 n0 min D psi Ce Flaw 0 000 mnin 100 0 DO 0 48 MS Method 2 File New 49 3 Total Run Time UPLC ErnErinnEnt setup cprojectmatsutuji pro acqudb upleonly Erp EE I x File EdiL options Toolbars Functions Help ms scan 4 SIR MRM Daughters Parents heura Loss Neutral Gan Surwen eed Total Hun Time 1 5 DD A 1Hmins Rs EE hs Scan Timg HH tn 15 H ass 1 HI HH tn 581 HH E 49 MSMethod S0 4 MS MS Scan SIR MS Scan SIR 50 MS Scan SIR Erperiment setup FtDFnjECEYTnaEsufuii nrniarnmnrhrninlrnnly Enh File Edit options Tnnlhars Functinn XX MR Daughters Parernts heutral Loss Meutral LE la LUrwe Tantal Hum Te 5 HI 1Imns Type Infarmation SS Scan Time HI tn 15 HH Wass 1 tn T1000 00 ES EE HS Scan
31. 3 Status Queue Ts Empty tatus Spnectrum Chramatngram Map Editr 5Samhles File Name File Text M5 File Inlet File Batlle Inlect Walume Cantral Ready DEFAULTOT Defautfle DEFAULT DEFAULT 10 000 EFALILTO2 Default flg DEFALULT DEFALULT 2 10 000 EF Default file DEFAULT DEFAULT 3 10 000 EFAILTOA4 Defaultrfie DEFAULT DEFA amp AULT A 10 000 pies EFALULTOS Default file DEFAULT DEFAULT 10 000 5 DEFAULTOE Default le DEFAULT DEFAULT 1a 5 10 000 7 DEFA amp ULTO Default file DEFAULT DEFAULT 1 4 7 10 000 nn eeliereir 38 2 Start Sample List Run Acquire Sample Datal Et 3 CIEE EN C MassLymsDEFAHLTPRO OK Pre Fum aITIhles cg amhle Data From 1 ntn Process Gaples File Name og uto uantity Samples a cheduling gt gt IPast Rumn Prinrity Night Time Process DE NDT 39 Start Sample List Run 23 Queuel 40 ile View Run Help Shortcut Queue Status Queue 1s Ermpty Queue Spectrum Chromatogram Map Editr 5Samples File Name File Text MS File Inlet File Bottle E
32. F 8 5 000 Pa Operate thod 29 Sample List ISamplesl IFormatl ILoad default fmt 30 WF MassLynr Matsufuji untitled File Wiew Run Help 1 gt NM rel hartcuk 3 Queue sat Dueue 15 EMmpty Spectrum hrnrmatn rarm Mabp Editr ams FNers eat MSFe Add me _ Tm Insert 0H0 a Dperate 0 halgtg n nn Fill k 8 Claar k Colurmn k ok OD Feady Format Fk CUstomize Sort k Load 5 0 Nurmhgr nf Samples 0 tn Furmber of Injections Tirng PFB55UTB In 5932 si UtnSampler Bed Lawnut Flom 30 Sample List 19 31 1 Hile Name 2 2 File Text 3 MS Hile UPLConly 4 Inlet Hile
33. Scan 0 ME 19193 hso oo 2m3463 Fm Ns scan 294 1 Fs Sca 2m 1 rs so hm 2034 53 5 100 MSMS MRM ipr vap915_MS25ETUP ip SiR MS 1pr vaB915_ESCI ipr vaB915_NEGSENS ipr vaB915_IDLE ipr AB915_POSSENS jpr vaB915_MS1HMRES ipr MS Scan MS 1pr WaB915_MS15ETUP jpr Daughter SCahn MSMS 1 pr ea ll 45 MS 27 MS COL GAS 0 mL min MS ipr APCI Options ME ion modes ESCi model 46 LT attro Prennier Cproje IniEi alLprovacqudb Mim File Ion Mode calibration Ge Ramps Options Help Inn ES Source Analyser Diagnosiics T WAVE Function Set Mass Span Gain Waltaes 1 ei le Hiz94 Ho Capillary kw ns EET 2 Scan MD Cane M _ E Extra
34. height areal hh a Reake tea am tnde Inmn ICE EriahiEhHIEE IIIEBSUTETIEI REspnnsE Threshnld Ln FEl absnlutm highi Fsak detect ApesTrack Peak Integration Relatiwe area ann Esanlule area Threshnld hromatoqram Rosemary_std 100_test I File Edit Display Process Window Tools Help x ov EE 0 A Glm es Std_100ugjmlLtest 12 Nov 2007 19 09 19 Rosemary_std 100 test Sm Mn 2x2 2 Diode Array 2 1 Range 3 33BE 1 1561156 Area 2 44 10133BB Ne 5 69 294353 387 45g1 3 3B5n 709 790 11219916 29045 30p5 3 31314 573082 11 00 Rnsemary_std 100 test 2 Diode Array Range 3 369e 1 3 0e 1 66 43 5 MS I X ectru File Edit Display Process Taos Windaow Help X ey Ce Tc TIC A GI 13 Nov 2007 14 07 59 Rosemary_solid_new_40ugiml 13 Nov 2007 14 07 59 1 Scan ES 67 O Rnsemary Snlidj newj 4Uudml 825 9 067 Cm 819 830 1 Scan ES 3 ES 329 18 1 57e6 100
35. 100 233 1 38e5 235 255 2 257 2 259 35 B9 77 87 99100 113121129 143 158 161 171 3 1g4196 199 214 222 gt 238 273 287289296309317 325332 334 343 5H H 100 120 140 160 180 200 220 240 2pH 2H qHH 320 340 5 4 59 MS IV LC MS MS 1 NN OO Q Pk Ww tM Inlet Method MS TUNE MS Method MS LC MS MS 1 Quattro Premier CiNprojecEinitialproacqudbnnrnmn ipr lox 1 File Ion Mode Calibration Gas Ramps Options Help E Ds SE gnostics T WAVE Function Set Mass Span Gain ayE 1 3 Scan lt 112 94 NN Save AS B44 so CD m ln EE DMS TUNE Fraperties D0 3 et ho 2 ii 9 hs i 100 Print CtrP 3 lm 60 Pin Previe MSMS ipr Ren File hi Print Setup Exit
36. 14 290 10 762 Cm 289 283 2 Scan ES 329 30 1 70e7 0 112 81 182 74 285 32 B31 39 397 30 473 90 921 88 990 05 0 test080214 291 10 789 Cm 289 294 1 Scan ES 331 13 9 69e7 100 683 20 328 22 684 15 2D 698 30 285 14 7039 As 62 116 95 178 93 440 21 531 13 82S 3 1 0 ee LE 1 i 100 200 300 400 500 600 700 800 900 46 MSMS Chromatogram Ai Std_mix high_ conc_MRMScan 25 5 313 Cm 21 27 A 9 Daughters of 331ES 100 287 23 2 86e5 271 18 203 29 104 45 177 00 189 8 215 18 269 35 328 80 Std_mix_high_conc_MRMScan 23 5 283 Cm 22 27 ee KN 8 Daughters of 331ES 400 271 18 4 57e4 215 06 203 05 243 10 201 20 189 02 176 96 _ 229 29 NN 44 09 148 97 174 90 n 111 99 138 21 NM Si 3 I 308 13_314 87 Std_mix_high_conc_MRMScan 24 5 304 Cm 21 27 Zs 6 7 Daughters of 331ES 80 201 11 215 18 271 21 1 76e4 243 20 227 18 122 84 287 12 6 i i 136 57 0 0 113 03 E 1 279 23 729 304 32 3
37. 2 MS INJECT er MS 54 INJECT MS 54 INJRECT 1 UPLC MS Hile 2 UPLC MS Hile UPLConly MS 3 IFilel Save Asl 55 MS Wa MassLynx souken souken3 5PL Fie yiew Run Help Open Project Project Wigard Oueue Is EMmpty Open Data File BMM Chromatogram 0 mM Samples v New Ctr N open crro la 1 Fest0802142 save Ctr 5 Save As oo Sample List Properties Import Worksheet Import Data eady gO Print CtrlTP Print 5etup A ee Exit ssure ro S Rn 55 HS3 UPLC 57 Wa MassL
38. 2008 03 13 UPLC MS MS kk 1 A A A GG 2 II UPLC kk 3 PC Gi 4 UE ON Gi Gc 4 oo 4 GE NM ON KKT 4 NO fb 5 PI OZ D C lil 6 oo 7 PDA EO ct i 8 MISC MCRO ON Cc 8 DACEONM RE CD ea a i i i A i i i 9 EO OR Gc 12 DAuto sampler Ok ENNED RE i i ik 13 Deteector kk N41 14 SMMS 16 oo 17 SR CA Dc 19 OG ik Gi 23 LC MS MS 26 i NR oc i ii ii 26 CC Ri i 26 eCEMEERGLO EN cc i cl 26 NISNIUNNINIOON 0 eo th NGA 26 APO ME i 28 MS 30 MS 82 LOW 2 32 NIS 33 2 MN i 33 GN i EE 34 INW MCAMSMMS 0 KMKO QNO 36 pi 36 DM TN OO cc Nl eA SD 36 OMSMeENGCCOAE Nc cc A 36 V kk 38 CO OR NN voi 38 DU ARNG NC ME ER OC 2 i 39 3 RC Sk 41 4 Of lk 43 SNMS OC a if fi 44 VI kk 48 WA cc Gc 48 ES RT A A cc i 48 OAPOI CRTUE EA Nk 48 OAPPI NG 0 KK 48 2
39. 27 90 _ 100 120 140 160 180 200 220 240 260 280 300 320 1 i i z 47 1 VI ESL M HI M NH4 M Nal IM KI IM HI IM CHsCOO1 IMTC1I APCI ESI NN SINT MR MTNH4 M nHlr MT NH2nlr M nHln M H M CH COQ APPTI 2 1 3 4
40. Bm teb EE Gradient Results Calculatel eS RS SD ou Diam Qeometrically Scaled UPLC Method Choices Hesults with Existin Column Peak Capacity 133 nom L 5caled haierk 2 1 mm Column Pressure bar 43 25 cm Scaled Gradient 1 0 mm Column Column Length 15 cm Qptimally Scaled UPLC Method Choices Column Diameter 4 6 mm Modifi 2 M Parlicle Size 5 pm Oraanic Modifi Maximum Peak Capacity at Enual Hun Time Mwr of Sample 300 Shortest amp nalysis Time at Equal Peak Capacity HPLC Linear Velocityl UPLC 9 view vem 1 2 NY 5 1 ee gm 94Wan tm ee Gradient Separations 16 EE Gradient Results OE dy ss Here are your UPLC methods Celu Dian PPometricaly Scaled UPLC Method Choices Hesults with Existing Column Peak Capacity 80 bh Scaled Gradient 2 1 mm Column N 30 Scaled Bradient 2 1 mm Lolumn a 25 5 4Um Saalad Cradiont 1 mm Polan ee hl 4 lt O 33m op Geometrically Scaled Gradient 2 1 X 100 mm x Organic Modifi Time Senment mm Imin Es Calculator Injection Volume 0 7 Pmax 343
41. Conly 25 MS Method UPLConly Total Run Time UPLC 0 00 Hilel Savel Erp tirnent setup CDFnTELE mmaEsuFuTi rnar udh unlrnnly Erp 1 x ptinons Tnnlhars Functions Help IX scmn 4 SIR MRMM Daughters EE Parents Neualt Tatal Hun Tme 5 HH 1Drmins NO 1 ME Scan Time OD to 15 HH hass 1HH HH tn 1 HH ES 25 MSMethod 16 4 Inlet Pump On 26 27 0 000 mL min
42. File Edit Display Process Window Tools Help mj A G ICo l e leAEI A GICGClme 9 Rosemary_ligud_new_200ug ml 13 Nov 2007 13 35 58 osemary_lidud_new_200ug ml 13 Nov 2007 13 35 58 1 2 IRasemary Liquid new 2 Hugml 11718 9 765 5 Dinde Array DSE nary LIiduid nEwij 20Hugrml 5 Dinde Array 3 n1e 2 1 15 Range 4 561 3 ne 2 c 2 75e 2 3 n 2 5e 2 2 5 2 25e 2 2 0 2 ne 2 1 5 1 75e 2 1 n 1 5e 2 kG 5 ne 1 1 25e 2 285 26 nn 1 HE 2 5 ne 1 7 5e 3 n nn 2 nn 12 nn 14 nn 5 HE 3 EnSBimary Liduidi nBwr 2UHu imli 1 Scan ES galas TIC 2 5e 3 2 82e8 nn Nm 9 07 10 12 ET 12 15 13 77 1121 33 on3 43 4 50 4 84 43 ca7 36 BBg0 lke _14 25 11 L Time n nn 2 nn 4 nn 6 nn 8 nn 10 00 12 00 14 00 TE 64 UV 59 filel IPrintl Rose test_05 15261 12 718 2 Diode Array 29980 4 07e 1 2 4 6 1 Rose_test 04 12791 10 660 2 Diode Array 3 627e 2 2 lt Rose_test 05 6437 5 364 2 Diode Array 329 30 2 119e 1 175e Se 1 5e 1 1 25e 1 N 1 0e 1 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400 40 3 IDisplay wavelengthl 65
43. bar 5 Inlet 17 Pressure Limits nihcaon Low hh psi Seal Wash lBz Io1zhconHMecN High 15000 ps Eo mn Comment 17 Inlet 11 5 Run time Binary Solyent Manager Instrument Metho _ e Binary Solvent Manager Fun Time soo min 18 5 gt KG a i Limits ni Han Low hh Seal Wyash lt nzHCnnHyMerN n Aigh 15000 ps En mi Gradient 0200 70 0 30 0 0200 20 0 8n n 6 1 0 200 20 0 80 0 6 9 1201 200 70 0 30 n 6 5 a Cancel Run Time 18 Run ime Data Analog Out Events 7 OK p8 11 12 Auto sampler General Run Time M
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47. ents Prime Set Defaults Start Cancel 5 Duration of prime 5 6 6 Seal wash Strong wash Weak wash Sample syringe 8cycles Start 5 GLalus AEHUITY Addiianal Status Flow e H5I Am2 5HH FI1 SN H HH mlmin UPLC Inlet method ACQUITY Additional Status 8 HP wyhp HP w2200 HP wpda Inlet Method File View Tools LC acquity Sampler Help Gtatus ACOQUITY Additional Status ACOUITY Binary 5olyent Manager ACQUITY Binary Solvent Manager A B 0 psi A1 100 0 Sa 25 8 C DO OO00 mLjmin B1 0 0 Off ALUUITY PDA L a Flow Communication failure autosampler Set Flow A B Solyent A Detector For Hel
48. ers ACOUITY UPLC Website kraHiEnt separations File Run Help bradient 5eparations DT aters Corporation Column Column Parhcle Hw of Temper lInjection Insktrument Lennth Diameter size 5 ample Flow Rate 5 mn g cm MM Hm nn a lt F lt Hrnarnic Modifier in B Pressure nits Hesults With Eishinn Column i a bar OP MPa Fe Capacitu Fessue ba ee mir Mus min ee DIUMmES mLmn IIH 1 n 1HH LO Init Cond Init Hold SB ZD7 waters Corporation Column of Fn hate ie Iniection Instrument Lenaqth Diameter Size Sample ature Volume Delay 30 cm lnim Ann 100 40 5 1 00 9m lt a mL mn Celsius ul mL Urqanic Modifier in B Pressure Units Hesults with Existing olumn MeCN J MenH bar Psl up PeakCapacty Fressure bar Time Time Flow Column SN min ImL min 4 ei Yolumes Init Cond IM 1 3H Ii Init Hold IO 1 IM Ii 20 00 1 0I 2 gD 20 00 12 15 3H I 1 0 2 gD 10D 00 E 8 DD 1 0 3H Ii Select Maximum Column Pressure EBHhar Chanae Click here for uour ACOUITY UPLC column choices This dialng box will display information as you click on each item s label 14 Calculator 10 Gradient Separations 15 Change Flow 0 2 0 4 Ee
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50. ndge 7 296e 2 6 0e 2 5 0e 2 4 0e 2 9 43 350 nm mm 3 0e 2 220 2 0e 2 10 67 0 1 0e 2 a 1 5 99 12 71 KW A 9 A BWS IE 0 00 2 00 4 00 6 00 8 00 10 00 12 00 14 00 test 080214 5 Diode Array 10 67 285 Range 2 434e 1 285 nm pk 2 5 37 i 13 09 6 00 14 00 5 Diode Array Range 5 944e 1 4 0e 1 N Be 12 71 3 0e 1 a ams 2 2 0e 13 1 0e 1 0 0 1 0e 1 Time 0 00 2 00 4 00 6 00 8 00 10 00 12 00 14 00 42 4 Processl Integratel 66 1ntegrate chromatogram OK Chromatogram Rosemary_std 100_test a la File Edit Display Process Window Tools Hep gt et Rm ete OA EEOC Kes 8 O Std_100ug mlLte smooth 12 Nov 2007 19 09 19 Subtrack Rnsemary_Std 1 Process Al Traces 2 Dinde Array Cambine Spectra Rande 3 36de 1 Components Signal To Noise Threshold Relative height areal Integrate chromatogram Absolute
51. ndify CQOUITYT Sample Manaeer Instrument Method Acouity Sample Manager Run Time 1 00 min ISample 5 19 Sample Lonp Option IP Laap Qffline PFartial Lnnp IDisable IT Partial Lann With Meedle Certill 1 Run Time Inlet Full Lnap 1 ONS s Wash Solwents Temperature Weak Cinlurmr 2 Column Pe Sample ef Sample 4 4 0 nmITEHt 19 Auto sampler 3 Sample loop option IPartial Loop With Needle Overfilll 4 Advancedl TE Advanced l Needle overfill frash 3 1 Data Event 5 OK p8 11 13 Detector 20 General Run Time Modify ACQUITY PDA Instrument Methnd Range
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