液质连用的基础Ultra Performance LC基础.pdf

?(UPLC?)?o?o?!?#?$?%?#?(?)?(?)?Chromatogram?(?)?&(?&)(?)?&*(?*?+*(?*?,?,?()2112WW21t tR+=()8.5.5211-5tR1-3=+=()2.5.5211-2tR1-2=+=t0t1t2t3?.5 1 2 5?()?+?=1N141R?-?3.5.5-2 k2=0V0V1V k1=:?1.5.5-1 k1=9.5.5-5 k3=V0V1V2V3?.5 1 2 5?k?0V1V0V2V 1k2k =?3.5-1.5-2 =12/?9 .5-1.5-5 =13/?V0V1V2V3?.5 1 2 5?.?1600 N.5516 NWV16N22=?=?=100N2516NWV16N22=?=?=PW=.5PW=2?/?)?0?/?*?0?o?o?o?o?o?ab5%?ab?1?2?1?3?2?3?ab)(a2+?#?4?5?6?0?)7?*0?7?4?7?*40)7?4?Waters HPLC?%B?!?8?+?0?9?)9?(:)?()6:/?Pressure vs.Organic-Water Percentage02004006008001000120014001600020406080100Percent Organic SolventpsiMeOHACN?2.1?30mm Symmetry C18?,20?Pressure vs.Flow Rate at 20?C05001000150020002500300035004000450050000.00.20.40.60.81.01.2mL/minpsi50%MeOH50%ACN100%H2O100%MeOH100%ACN?Pressure vs.Temperature050010001500200025003000350040004500500010.020.030.040.050.060.070.0Degrees,Cpsi50%MeOH50%ACN100%H2O100%MeOH100%ACN?(?)?6?*?;)?3574 5 6?)?4=?,?)7?+?A?)?)?B 9?2?3?Retentivity(ln k acenaphthene)SunFireC18YMC-PackPolymerC18HypersilCPS CyanoYMC-PackCNWaters Spherisorb S5 PHypersilBDS PhenylNova-PakPhenylYMC-PackPhenylHypersilPhenylInertsilPh-3YMC-PackPro C4YMCbasicSymmetryC8YMC-PackPro C8Nova-PakC8XTerraMS C18SymmetryC18YMC-PackPro C18InertsilODS-3YMC-Pack ODS-ANova-PakC18YMC J sphereODSL80NucleosilC18Waters SpherisorbODS2Waters SpherisorbODS1ResolveC18BondapakC18YMC-PackODSAQYMC J sphereODSH80YMC J sphereODSM80InertsilCN-3Waters SpherisorbS5CNNova-PakCN HPSymmetryShieldRP8SymmetryShieldRP18XTerraRP8XTerraRP18-0.6-0.300.30.60.91.21.51.82.12.42.733.33.6-1.5-0.50.51.52.53.5Selectivity(ln amitriptyline/acenaphthene)XTerraMS C8LunaC18(2)ACQUITY UPLCBEH XBridgeC18XTerraPhenylLunaPhenyl HexylChromolithRP-18AtlantisdC18ZorbaxXDB C18ACT Ace C18ZorbaxSB C18SunFireC8LunaC8(2)ACQUITY UPLC BEHXBridgeShield RP18ACQUITY UPLCBEH XBridgeC8ACQUITY UPLCBEHXBridgePhenyl?ACQUITY UPLC0.080 0.090?Parker/Swagelok?Waters?0.120 0.130?Waters?0.009?C?D?D?10 min?3.5-5m?1500-4000 psi(110-280 bar)?80,000-115,000/?10m?1000-2500 psi(70-180 bar)?40,000/?10 min?40m?100-500 psi(7-40 bar)?5000/?10 min?*50 mm?*?2.0 mL/min0.0123.0 mL/min.12Time in Minutes3.0?Rs=3?(?)5 m?3321.,=Rs3221.,=Rs?E?=?2468101214161820222426283000.511.522.533.54?mm/secH(m)5 m?2.5 m?(50/50,?/?)HETP?%1?H=HETP(Height Equivalent to Theoretical Plate)?H?,?N=#Plates(?)5?10?(?H)(?H)“?”“?”L?HETPL N=PlateHETP?E?=?A?E?=?HETP?HETP=?mm/secC?(?)B?(?)C?B?dpcbdpaH2)()(+=?E?=?Height Equivalent to Theoretical Plate HETP?umm/secu=L/t 0HETP PlatesHETP PLATESH?10000500010001.02.03.0?mL/min?N?0.150.041?0.30.072?0.60.134?0.90.206?1.20.288?1.50.3410?!?#$?%?!?#$?$?&LC/MSu(mm/s)2.1 x 20 mm,1.7 m2.1 x 20 mm,2.5 m2.1 x 20 mm,5 m024681012140.0002.0004.0006.0008.00010.00012.000HETP(m)UPLC Range“Narrow-bore”?E?=?5m x 4.6mm ID =1mL/min3.5m x 4.6mm ID =1.7mL/min?1970s 30+min.1980s10-15 min.1990s5-10 min.2004 1-2 min.58,820 m 1.7mm100 29,500 m 1.7mm50 33,000 m3 mm100 30,000 m 5mm150 30,000 m10 mm300=2x?=dLp?0?AU0.000.020.040.060.080.000.501.001.502.002.503.003.504.004.50AU0.0000.0100.0200.005.0010.0015.0020.0025.0030.0035.00AU0.000.020.040.060.001.002.003.004.005.006.007.008.009.009.50AU0.000.050.100.15Minutes0.00 0.10 0.200.30 0.40 0.50 0.600.70 0.80 0.901.001.1010 m 250 mmRs(2,3)=1.545 m 150 mmRs(2,3)=2.693.0 m 100 mmRs(2,3)=2.291.7 m 50 mmRs(2,3)=2.25AU0.000.050.100.15Minutes0.000.100.200.300.400.500.600.700.800.901.00AU0.000.050.100.15Minutes0.000.501.001.502.002.503.003.504.00AU0.000.050.100.15Minutes0.001.002.003.004.005.006.007.008.00AU0.000.050.100.15Minutes0.005.0010.0015.0020.0025.0030.001.104.509.5035.00?Y-?L/dp?0?10531.7124.225.864.935.9231.542.692.292.2547.4610.049.1110.3?(m)?10531.711346534883419368608722431665209768851536253241846446877871152450413487351444623385613?(m)?tg-0.0050.0000.0050.0100.0150.0200.0250.0300.0350.000.100.200.300.400.500.600.700.800.901.001.101.201.301.401.50wwContaminant PeakwwwtPg+=1?w=?PW?3?F?#1 G?E?=?3?F?#1 G?AU0.000.100.20Minutes0.050.100.150.200.250.300.350.400.450.500.550.600.000.020.040.060.080.100.120.140.16Minutes0.000.501.001.502.002.503.003.504.004.505.005.506.001.Thiourea-0.4302.toluene-1.0343.propylbenzene-1.7424.butylbenzene-2.4135.hexylbenzene-5.0580.180.200.220.24HPLC?:52.2 m0.7 mL/min?:6.00?1.Thiourea-0.0462.toluene-0.0883.propylbenzene-0.1374.butylbenzene-0.1825.hexylbenzene-0.360?:51.7 m1.4 mL/min?:0.60?2?,10 x?0.6?6.0?(0.85 mL?4.2 mL)UPLCTM?5.0 m193001.7 m44165MinutesAU0.0000.0100.0200.0300.0400.0503.003.504.004.505.005.506.006.507.007.508.000.25AUMinutes0.01.6?:3X(300%)?:9X(900%)?:1.7(70%)?AU0.000.020.040.060.080.10Minutes0.00 51015202530354045505560PC=360?1234ACQUITY UPLCTMBEH C182.1 x 50 mm,1.7 mF=0.3 mL/minPSIMAX=4,200T(4)=1.63N(4)=5,400Rs(2,3)=0.97XTerraMS C182.1 x 50 mm,2.5 mF=0.5 mL/minPSIMAX=3,950T(4)=1.30N(4)=4,100Rs(2,3)=1.10AU-0.0020.0000.0020.0040.0060.0080.0100.0120.0140.0160.0180.0200.022Minutes0.001.002.003.004.00AU-0.0020.0000.0020.0040.0060.0080.0100.0120.0140.0160.0180.0200.022Minutes0.001.002.003.004.00?3?F?#1 G?1234AU0.000.020.040.060.080.100.120.140.16Minutes0.000.501.001.502.002.503.00ACQUITY UPLCTMBEH C182.1 x 50 mm,1.7 mAlliance2695F=0.3 mL/minPSIMAX=4,200T(4)=1.63N(4)=5,400Rs(2,3)=0.97ACQUITY UPLCTMBEH C182.1 x 50 mm,1.7 mACQUITY UPLCTMF=0.6 mL/minPSIMAX=8,400T(4)=1.02N(4)=10,100Rs(2,3)=2.25 AU0.000.020.040.060.080.100.120.140.16Minutes0.000.501.001.502.002.503.00HPLC?UPLC?3?F?#1 G?10.0Rs=1.86Rs=2.302.1x100mm 5 Hybrid C18Time in Minutes0.010.0Rs=3.01Rs=6.452.1x100mm 5 Hybrid C18ACQUITY(HPLC)HPLC12?3?F?#1 G?1 0.25AUFS10.0Rs=3.01Rs=6.452.1x100mm 5 Hybrid C18ACQUITY(HPLC)0.30AUFSTime in Minutes0.010.0Rs=4.71Rs=9.152.1x100mm 1.7 Hybrid C18?,?ACQUITY UPLCTM23?3?F?#1 G?1 10.00.30AUFSRs=4.71Rs=9.152.1x100mm 1.7 Hybrid C180.33AUFSTime in Minutes0.03.5Rs=3.52Rs=1.822.1x30mm 1.7 Hybrid C18?HPLC?,?ACQUITY UPLCTMACQUITY UPLCTM34?3?F?#1 G?1 0.33AUFSTime(min)0.03.52.1x30mm 1.7 Hybrid C18?HPLC?,?1.60.25AUFSTime(min)0.02.1x30mm 1.7 Hybrid C18?1.60.25AUFSTime0.0ACQUITY UPLCTM45?10.02.1x100mm 5 Hybrid C18ACQUITY(UPLC)TMHPLC1.60.25AUFSTime(min)0.0152.1x30mm 1.7 Hybrid C18?/?C?9%?):?C?AU0.0000.0050.0100.0150.001.002.003.004.005.006.007.008.009.000.3 mL/min.AU0.0000.0050.0100.0150.001.002.003.004.005.006.007.008.009.000.5 mL/min.AU0.0000.0050.0100.015Minutes0.001.002.003.004.005.006.007.008.009.000.7 mL/min.2.3X?B 1?*:)?/?/:?)?6?3?F?#1 G?3?F?#1 G?9%?)C?3?F?#1 G?):)?0?B 1?o?H5/?AU0.0000.0100.0200.030Minutes0.000.200.400.600.801.001.201.401.601.802.002.202.402.602.803.000.4590.5890.7450.9661.4191.7782.653?AU0.000.020.040.06AU0.000.020.040.06AU0.000.020.040.06Minutes0.000.501.001.502.002.503.003.504.004.505.00Peak1Peak2Peak3Peak4Peak5Peak6Peak7B 1?3?F?#1 G?XTerra 2.1 x 50mm 2.5 m.4.0?,0.5 mL/min.ACQUITYUPLC BEH C182.1 x 50mm 1.7 m.4.0?,0.5 mL/min.ACQUITY UPLC BEH C182.1 x 50mm 1.7 m.0.8?,1.0 mL/min?AU0.0000.0050.0100.0150.0200.0250.0300.0350.0400.045Minutes0.000.501.001.502.002.503.003.504.004.505.00Peak1Peak2Peak3Peak4Peak5Peak6Peak7B 1?*:)?/?3?F?#1 G?(?):1.?2.?3.?4.?5.?6.?7.?3?F?#1 G?9%?)?C?3?F?#1 G?AU0.0000.0050.0100.0150.0200.0250.0300.0350.0400.0450.0500.0550.060Minutes0.000.100.200.300.400.500.600.700.800.901.001.101.201.301.401.50Peak1Peak2Peak3Peak4Peak5Peak6Peak7?(?):1.?2.?3.?4.?5.?6.?7.?AU0.000.020.04AU0.000.020.04AU0.000.020.04Minutes0.000.100.200.300.400.500.600.700.800.901.001.101.201.301.401.50HPLCUPLC5 m?(17 L)?=361.7 m?(0.5 L)?=802.1 x 50 mm ACQUITY UPLC?,1.0 mL/min,?0.8?,5%?35%ACN5 m?(4 L)?=46HPLCRs=1.4Rs=1.8Rs=2.5?AU0.0000.0100.0200.0300.0400.050Minutes0.600.801.001.201.401.601.805 m HPLC boreAU0.000.020.04Minutes0.500.600.700.800.901.001.101.201.301.401.501.601.7 mUPLC?5 mHPLC?Alliance?48?Rs=1.4Rs=1.8Rs=2.5AU0.0000.0100.0200.030Minutes0.400.600.801.001.201.401.601.802.00Rs?1.8?1.7?Rs?1.3?2.4?*?C?3?F?#1 G?