液質(zhì)聯(lián)用實(shí)用PPT學(xué)習(xí)教案

1、會(huì)計(jì)學(xué)1液質(zhì)聯(lián)用實(shí)用液質(zhì)聯(lián)用實(shí)用第1頁(yè)/共76頁(yè)自動(dòng)進(jìn)樣器 色譜柱 PDA（陣列二極管檢測(cè)器） 離子化室 質(zhì)量分析器檢測(cè)器液相色譜 質(zhì)譜計(jì)算機(jī)第2頁(yè)/共76頁(yè)質(zhì)譜質(zhì)譜離子化室離子化室第3頁(yè)/共76頁(yè)四元泵四元泵(包括包括真空脫氣機(jī)真空脫氣機(jī))柱溫箱柱溫箱PDA檢測(cè)器檢測(cè)器溶劑瓶箱溶劑瓶箱自動(dòng)自動(dòng)進(jìn)樣器進(jìn)樣器第4頁(yè)/共76頁(yè)液相色譜流程圖液相色譜流程圖PUMPDetectorWData station (Recorder) or Collection第5頁(yè)/共76頁(yè)第6頁(yè)/共76頁(yè)第7頁(yè)/共76頁(yè)第8頁(yè)/共76頁(yè)第9頁(yè)/共76頁(yè)第10頁(yè)/共76頁(yè)混合器混合器B泵泵A泵泵梯度程序控制梯度程序控制第

2、11頁(yè)/共76頁(yè)混合器混合器高壓輸液高壓輸液四通四通第12頁(yè)/共76頁(yè)第13頁(yè)/共76頁(yè)第14頁(yè)/共76頁(yè)第15頁(yè)/共76頁(yè)第16頁(yè)/共76頁(yè)第17頁(yè)/共76頁(yè)第18頁(yè)/共76頁(yè)第19頁(yè)/共76頁(yè)在強(qiáng)電場(chǎng)下溶液帶電在強(qiáng)電場(chǎng)下溶液帶電 形成帶電液滴形成帶電液滴帶電液滴體積不斷縮小帶電液滴體積不斷縮?。ㄈ軇┱舭l(fā)，不斷發(fā)生庫(kù)侖爆炸）（溶劑蒸發(fā)，不斷發(fā)生庫(kù)侖爆炸） 離子逸出表面，蒸發(fā)進(jìn)入空間離子逸出表面，蒸發(fā)進(jìn)入空間電噴霧電離的實(shí)質(zhì)：電荷濃縮效應(yīng)電噴霧電離的實(shí)質(zhì)：電荷濃縮效應(yīng)電噴霧電離的實(shí)質(zhì)：電荷濃縮效應(yīng)電噴霧電離的實(shí)質(zhì)：電荷濃縮效應(yīng)第20頁(yè)/共76頁(yè)+_+_+ +蒸發(fā)蒸發(fā) 帶電液滴帶電液滴小液滴小液

3、滴 ESIESI離子蒸發(fā)機(jī)理離子蒸發(fā)機(jī)理第21頁(yè)/共76頁(yè)第22頁(yè)/共76頁(yè)第23頁(yè)/共76頁(yè)第24頁(yè)/共76頁(yè)第25頁(yè)/共76頁(yè)第26頁(yè)/共76頁(yè)第27頁(yè)/共76頁(yè)第28頁(yè)/共76頁(yè)090905-taoyang-3 #39RT: 0.55AV: 1NL: 1.44E7T: + c Q1MS 400.00-1500.004005006007008009001000110012001300m/z05101520253035404550556065707580859095100Relative Abundance605.08583.09606.08621.07662.26563.19663.265

4、03.10767.281081.65855.001187.72896.701326.69966.00M+Na+M+K+M+H+MW=582第29頁(yè)/共76頁(yè)第30頁(yè)/共76頁(yè)第31頁(yè)/共76頁(yè)QOOQOQ1HyperquadQ3HyperquadQ2Collision CellDetectionSystemIon SourceInterfaceHeatedCapillaryMass Analyzer第32頁(yè)/共76頁(yè)Q1Q3Q2Q0DynodeTurbo第33頁(yè)/共76頁(yè)第34頁(yè)/共76頁(yè) 為了區(qū)分這四個(gè)離子，測(cè)量的質(zhì)量的準(zhǔn)確度需達(dá)為了區(qū)分這四個(gè)離子，測(cè)量的質(zhì)量的準(zhǔn)確度需達(dá)到到130 ppm。

5、即小數(shù)點(diǎn)后。即小數(shù)點(diǎn)后第四位是準(zhǔn)確的第四位是準(zhǔn)確的。 元素組成信息隨著質(zhì)量的增加而呈指數(shù)地上升，元素組成信息隨著質(zhì)量的增加而呈指數(shù)地上升，對(duì)質(zhì)譜質(zhì)量的測(cè)定準(zhǔn)確度要求更高。對(duì)質(zhì)譜質(zhì)量的測(cè)定準(zhǔn)確度要求更高。 12C = 12.00000000 ， 1H = 1.007782506， 14N = 14.00307407， 16O = 15.99491475 m=43.0184Da的離子的離子必定是必定是C2H3O+，不可能是不可能是C3H7+(m=43.0058)或或 C2H5N+(m=42.9984)，或或CH3N2+(m=43.0296)。第35頁(yè)/共76頁(yè)第36頁(yè)/共76頁(yè)第37頁(yè)/共76頁(yè)第

6、38頁(yè)/共76頁(yè)第39頁(yè)/共76頁(yè)Flow Rate Column I.D.1.0 mL/min 4.6 mm 0.5 mL/min 3.0 mm 0.2 mL/min 2.1 mm 50 mL/min 1.0 mm 10 mL/min CapillaryLC流速一般根據(jù)流速一般根據(jù)色譜柱規(guī)格色譜柱規(guī)格和和電離方式電離方式來(lái)確定來(lái)確定APCI 0.2-2 ml/minESI 5-300 l/min routinely (1 ml/min possible)第40頁(yè)/共76頁(yè)第41頁(yè)/共76頁(yè)第42頁(yè)/共76頁(yè)第43頁(yè)/共76頁(yè)Full-Scan Mass SpectrometrylAdvant

7、agelProvides MW InformationlDisadvantagelLow duty cycleScanningPass AllPass All第44頁(yè)/共76頁(yè)Full-Scan MS of BuspironeNNNNNOOBuspirone(丁螺環(huán)酮丁螺環(huán)酮 )C21H31N5O2MW = 385150200250300350400450500m/z255075100Relative Abundance386408(M+H)+(M+Na)+第45頁(yè)/共76頁(yè)Single Ion Monitoring (SIM)lAdvantageslTargeted Analyte Moni

8、toringlHigh Duty CyclelSimplelDisadvantageslCan suffer from interferenceslNot as sensitive or selective as SRM (see below)Fixed m/zPass AllPass All第46頁(yè)/共76頁(yè)Selected Reaction Monitoring(SRM)lAdvantageslTargeted Analyte MonitoringlHigh Duty Cyclel“Simultaneous” Monitoring of Multiple TransitionslDisad

9、vantagelNo “advanced” structural informationFixed m/zPass AllFixed m/zQ1Q2Q3第47頁(yè)/共76頁(yè)清洗儀器及管路配樣 （稀釋?zhuān)┵|(zhì)譜條件優(yōu)化（保存方法）液質(zhì)聯(lián)用方法設(shè)置平衡色譜柱分析結(jié)果進(jìn)樣品結(jié)果無(wú)效第48頁(yè)/共76頁(yè)第49頁(yè)/共76頁(yè)液質(zhì)聯(lián)用與氣質(zhì)聯(lián)用的區(qū)別液質(zhì)聯(lián)用與氣質(zhì)聯(lián)用的區(qū)別第50頁(yè)/共76頁(yè)第51頁(yè)/共76頁(yè)第52頁(yè)/共76頁(yè)第53頁(yè)/共76頁(yè)C:Documents and Settings.s045/27/2009 4:49:53 PMRT: 3.66 - 31.3451015202530Time (min)01

10、0000002000000uAU0200000uAU05010018.4218.4919.4120.6626.4430.1924.204.2128.285.687.1914.4810.778.8812.4917.4318.2522.716.184.9125.347.029.4218.2622.726.187.0225.483.999.4221.6727.54NL:1.29E8TIC F: MS s04NL:3.43E5Total Scan PDA s04NL:2.41E6Channel A UV s04s04 #1063RT: 18.44AV: 1NL: 6.10E7T: + c Q1MS 1

11、20.00-900.00200300400500600700800900m/z0102030405060708090100Relative Abundance412.97414.98416.03588.27279.05475.07847.18701.30239.98338.11146.13605.14814.43887.44準(zhǔn)分子離子總離子流圖液相色譜圖全掃描單波長(zhǎng)掃描一級(jí)質(zhì)譜圖第54頁(yè)/共76頁(yè)090527 #778RT: 9.56AV: 1NL: 1.74E7T: + c Full ms2 413.00-22.00 30.90-420.0050100150200250300350400m/

12、z05101520253035404550556065707580859095100Relative Abundance86.09124.96339.87293.91412.9874.12227.9458.17258.95124.41327.82341.90182.00199.82127.05367.20總離子強(qiáng)度質(zhì)量范圍母離子質(zhì)量檢測(cè)離子模式碰撞能量母離子碎片離子二級(jí)質(zhì)譜第55頁(yè)/共76頁(yè)第56頁(yè)/共76頁(yè)第57頁(yè)/共76頁(yè)010203040020406080100Relative abundancem/z2022第58頁(yè)/共76頁(yè)元素AA+1A+2元素類(lèi)型質(zhì)量% 質(zhì)量%質(zhì)量%H110020

13、.015AC12100131.1A+1N14100150.37AO16100170.04180.20A+2F19100ASi28100295.10303.40A+1,A+2P31100AS32100330.80344.40A+2Cl351003732.5A+2Br791008198.0A+2I127100A第59頁(yè)/共76頁(yè)32! 3)2)(1(! 2) 1(1)1 (bnnnbnnnbbnb: A+1或A+2同位素的天然豐度n: 質(zhì)譜峰中A+1或A+2元素的個(gè)數(shù)A+1A+1元素峰位置元素峰位置M MM+1M+1M+2M+2M+3M+3A+2A+2元素峰位置元素峰位置M MM+2M+2M+4M

14、+4M+6M+6質(zhì)譜峰強(qiáng)度質(zhì)譜峰強(qiáng)度1 1nbnbn n( (n-n-1)1)b b2 2/2!/2!n n( (n n-1)(-1)(n n- -2)2)b b3 3/3!/3!第60頁(yè)/共76頁(yè)MM+2M+4M+6MM+2M+4M+2MBrBr3Br210010010033.310050509833.3第61頁(yè)/共76頁(yè)MM+2M+4M+6MM+2M+4M+2MClCl3Cl210010010010.631.765.032.53.497.5例：例：35Cl 100%, 37Cl 32.5% M M+2 M+4 M+6 Cl, n=1: 1 0.325 Cl2, n=2: 1 0.650 0

15、.106 Cl3, n=3: 1 0.975 0.317 0.034 第62頁(yè)/共76頁(yè)第63頁(yè)/共76頁(yè)090905-taoyang-3 #39RT: 0.55AV: 1NL: 1.44E7T: + c Q1MS 400.00-1500.004005006007008009001000110012001300m/z05101520253035404550556065707580859095100Relative Abundance605.08583.09606.08621.07662.26563.19663.26503.10767.281081.65855.001187.72896.7013

16、26.69966.00M+Na+M+K+M+H+MW=582第64頁(yè)/共76頁(yè)090222-cl-4h #17RT: 0.15AV: 1NL: 1.01E8T: + c Q1MS 460.00-900.10500550600650700750800m/z05101520253035404550556065707580859095100Relative Abundance539.10573.07540.12575.07576.08541.13537.70607.04569.70610.04535.72542.15636.13473.14659.13693.22715.25751.21794.05

17、M+H+NNHNHNH3CCH3CH3H3CCOOCH3COOCH3ClM=572第65頁(yè)/共76頁(yè)090512-2.5 #11RT: 0.13AV: 1NL: 2.12E6T: + c Q1MS 450.00-950.10580600620640660680700720740760m/z05101520253035404550556065707580859095100Relative Abundance619.02696.94620.06698.94694.94596.44621.09700.01584.45688.34645.11605.15701.26640.46732.45661.11

18、 675.04744.43760.46NNHNHNH3CCH3CH3H3CCOOCH3COOCH3BrBrNNHNHNH3CCH3CH3H3CCOOCH3COOCH3BrM=616M=694第66頁(yè)/共76頁(yè)080607-DPDME #40RT: 0.46AV: 1NL: 1.50E7T: + c Full ms2 539.00-40.00 30.00-540.00300320340360380400420440460480500520540m/z05101520253035404550556065707580859095100Relative Abundance539.11393.03407

19、.06465.06379.01408.05466.10451.05409.03378.03507.07479.12365.04524.10348.98312.23-CH2COOCH3 -COOCH3 -CH2-CH2-CH2-C=ONHNHNNCH3CH3COOCH3COOCH3H3CH3C第67頁(yè)/共76頁(yè)090625-599 #39RT: 0.50AV: 1NL: 7.58E6T: + c Full ms2 599.00-45.00 300.00-610.00300350400450500550600m/z05101520253035404550556065707580859095100R

20、elative Abundance511.11555.15599.13452.11537.10496.13495.11451.11581.13478.10437.10393.10379.09525.04407.04 423.14569.06378.02359.99323.88603.98NHNHNNCOOHCOOHHOOH-H2O -CHOHCH3 -H2O-CHOHCH3-CH3第68頁(yè)/共76頁(yè)100308-tuotie #49RT: 0.54AV: 1NL: 2.62E7T: + c Full ms2 511.00-45.00 150.00-530.0034036038040042044

21、0460480500520m/z05101520253035404550556065707580859095100Relative Abundance393.06379.03452.07437.04407.05378.02511.08377.05362.96433.11493.05348.92419.04465.13483.53516.48-CH2COOH-CH3-2CH3 -CH2-CH2 NHNHNNCOOHCOOH第69頁(yè)/共76頁(yè)100703-Mn-yuanbulin #30RT: 0.32AV: 1NL: 1.46E7T: + c Full ms2 615.00-45.00 300.

22、00-640.00300350400450500550600m/z05101520253035404550556065707580859095100Relative Abundance556.00496.99614.92541.08510.98496.05597.07481.03538.12578.14453.28427.94331.50625.03385.51364.24NNNNCOOHCOOHMn-CH2COOH-CH2COOH第70頁(yè)/共76頁(yè)10-10-26-liuxiaopeng #91RT: 1.12AV: 1NL: 3.39E7T: + c Q1MS 100.00-500.001

23、00150200250300350400450500m/z05101520253035404550556065707580859095100Relative Abundance322.04344.02277.07359.99299.04274.19376.03197.13437.06453.05402.89454.04150.99246.18198.13130.15487.35172.14原料原料+H+原料原料+Na+M+H+M+Na+M+K+第71頁(yè)/共76頁(yè)10-10-26-liuxiaopeng #31RT: 0.40AV: 1NL: 1.77E7T: + c Full ms2 322.00-25.00 30.00-350.0050100150200250300350m/z05101520253035404550556065707580859095100Relative Abundance321.96100.05128.05276.02102.0898.1170.07218.96206.8444.16275.27114.02178.91277.94148.89245.24320.06322.66CNNO2ONOH3CO+H+CHNHONOH3COCH2+NOCH3+NO實(shí)例實(shí)例9第72頁(yè)/共76頁(yè)100515-renqian