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1、,Chapter Five High Performance Liquid Chromatography For Short : HPLC,第五章 高效液相色谱法,高效液相色谱,高效液相色谱法 HPLC High Performance Liquid Chromatography,高效液相色谱法 HPLC High Performance Liquid Chromatography,往复式高压恒流泵,高效液相色谱法 HPLC High Performance Liquid Chromatography,注射泵,高效液相色谱法 HPLC High Performance Liquid Chrom
2、atography,梯度洗脱（淋洗）,低压梯度高压梯度,浓度,时间,高效液相色谱法 HPLC High Performance Liquid Chromatography,正相色谱 反相色谱 Reverse-phase Chromatography,液固色谱 Liquid-Solid Chromatography,液固吸附色谱 Liquid-Solid Chromatography,m,ad,ad,m,nM,X,nM,X,+,+,以样品中各组分在固定相上的吸附作用不同来进行分离,“相似相溶”原理异构体的分离,液液分配色谱 Liquid-Liquid Chromatography,Ki,Stat
3、ionary phase,Mobile phase,固定液易流失 化学键合固定相,化学键合相色谱 Bonded Phase Chromatography,1 功能基团的引入方法,S,O,C,l,2,C,l,R,M,g,C,l,R,S,i,O,H,S,i,C,l,S,O,C,l,2,R,M,g,C,l,S,i,R,化学键合相色谱 Bonded Phase Chromatography,化学键合相色谱 Bonded Phase Chromatography,R,-C4, -C8, -C18-C6H5, -CN, -NH2-SO3H, -COOH, -NR4Cl-,化学键合相色谱 Bonded Ph
4、ase Chromatography,正相色谱 3 反相色谱 RP-HPLC 4 反相离子对色谱 RP-Ion-Pair HPLC,反相离子对色谱 RP-Ion-Pair HPLC,Chromatogram of Mg, Cd and Zn separation,化学键合相色谱 Bonded Phase Chromatography,5 离子交换色谱,化学键合相色谱 Bonded Phase Chromatography,化学键合相色谱 Bonded Phase Chromatography,空间排斥（阻）色谱 Size Exclusion Chromatography,硅胶型、聚苯乙烯型、葡
5、聚糖凝胶型,空间排斥（阻）色谱 Size Exclusion Chromatography,M.W.,tR,空间排斥（阻）色谱 Size Exclusion Chromatography,M.W.,tR,空间排斥（阻）色谱 Size Exclusion Chromatography,M.W.,tR,空间排斥（阻）色谱 Size Exclusion Chromatography,M.W.,tR,空间排斥（阻）色谱 Size Exclusion Chromatography,M.W.,tR,Conditions: Column, Vydac C18, 5-mm, 30 nm (1 mm ID100
6、 mm)Flow rate, 40 mL/minMobile phase, A: 0.05% Formic Acid in WaterB: 0.05% Formic Acid in AcetonitrileLinear gradient programmed from 20% B to 100% B over 25 min,hold at 100% B for 5 min Sample: mixture of 9 protein standards; 5-mL loop Detection: SCIEX TAGA 6000E triple quadrupole mass spectrometr
7、yequipped with ionspray atmospheric pressure ion source inpositive ion mode 1, substance P(1-9),MW=1104, 900 pmoles; 2, neurotensin, 1673, 150; 3, angiotensin, 1296, 385; 4, somatostatin, 1638, 600; 5, bovine insulin, 5732, 130; 6, bovine glucagon, 3482, 720; 7, mellitin, 2846, 175; 8, myo- Globin,
8、16950, 20; 9, lactoglobulin-B, 18277, 245.,m-HPLC-Mass Spectrometry of Peptides and Proteins,m-HPLC-Mass Spectrometry of Peptides and Proteins,Mass Spectrum of Substance P from Peak 1,Mass Spectrum of Bovine Insulin from Peak 5,Mass Spectrum of Bovine Glucagon (胰增血糖素）from Peak 6,Mixed Mass Spectrum
9、from Peak 8,Separation and detection of the pigments in narcissus leaves by m-HPLC-APCI-MS,Separation and detection of the pigments in narcissus leaves by m-HPLC-APCI-MS,Separation and detection of the pigments in narcissus leaves by m-HPLC-APCI-MS,Separation and detection of the pigments in narcissus leaves by m-HPLC-APCI-MS,Separation and detection of the pigments in narcissus leaves by m-HPLC-APCI-MS,Separation and detection of the pigments in narcissus leaves by m-HPLC-APCI-MS,Separation and detection of the pigments in narcissus leaves by m-HPLC-APCI-MS,
