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中药复方小柴胡汤首见于东汉张仲景的《伤寒杂病论》,主治少阳症,由柴胡Radix Bupleuri、黄芩 Scutellaria baicalensis、人参 Panax ginseng、半夏 Rhizoma Pinelliae preparata、炙甘草 honey fried Glycyrrhizaew、生姜Zingiber officinale和大枣Zizuiphus jujuba七味药组成。现代药理研究表明,小柴胡汤具有治疗慢性肝炎、抗抑郁等作用,越来越多地受到人们的关注。然而小柴胡汤的抗抑郁活性成分仍不明确,本文在综述小柴胡汤化学成分和药理活性研究的基础上,首先利用D101型大孔吸附柱色谱将小柴胡汤水提物洗脱(乙醇-水)为四个部分:水洗(XCHD-1),30%乙醇洗(XCHD-2),60%乙醇洗(XCHD-3)和 95%乙醇洗(XCHD-4)。用抗抑郁作用的相关参数对这四个部分的活性进行评价,对有效活性部位XCHD-3运用正反相柱色谱,CHP20柱色谱、凝胶柱色谱、制备薄层以及分析、制备液相等多种色谱分离手段,从小柴胡汤活性部位中分离得到44个化合物,通过理化方法和波谱解析等方法鉴定了其中40个化合物的结构,主要为黄酮及其苷类化合物,分别为:isoliquiritin(1),neoisoliquiritin(2),5,7,2’,6’-tetrahydroxy-dihydroflavonol(3),chrysin-8-C-β-D-glucopyranoside(4),liquiritin(5),5,2’,6-trihydroxy-7,8-dimethoxyflavone 2’-O-β-D-glucopyranoside(6)、viscidulin Ⅲ(7),baicalin(8),luteolin-7-O-gentibioside(9),5,7-dihydroxy-6-methoxyflavone-7-glucuronide methyl ester(10),viscidulin Ⅰ(11),3,5,7,2’6’-pentahydroxy flavone(12),wogonoside-7-O-β-D-glucuronide methyl ester(13),wogonoside(14),glucoliquiritin apioside(15),wogonin(16),6,7-dimethoxy-4-hydroxy-1-naphthoic acid(17),myricetin(18),quercetin(19),martynoside(20),5,6’-dihydroxy-7,8-dimethoxy-2’-O-β-D-glucopyranosylflavone(21),ononin(22),syringing(23),3-(4-hydroxyphenyl)propionic acid methyl ester(24),apiol(25)chrysin-6-C-glucosyl-8-C-arabonoside(26),chrysin-6-C-arabinosyl-8-C-glucoside(27),liquiritigen-7-O-β-D-glucoside(28),apigenin 7,4’-O,O-diglucoside(29),3-isobutylpyrrolopiperazine-2,5-dione(30),vicenin-2(31),apigenin 6,8-di-C-arabinopyranoside(32),rutin(33),scutellarin(34),schafloside(35),isoschafloside(36),kaempferol-3-O-β-D glucopyranoside(37),quercetin(38),kaempferol(39),vanillic acid(40).利用分离到的已知化合物及购买的标准品为对照品,通过对比保留时间、总结已知化合物的质子裂解碎片规律以及文献查阅,对小柴胡汤全方及D101大孔树脂各洗脱部位(XCHD-1,2,3,4)采用LC-ESI-Q-TOF-MS/MS正负离子模式进行定性鉴别,结果共检测到包括多糖(inulin、sucrose,fructose/glucose,stachyose),黄 酮(liquiritigenin-7,4’-diglucoside,liquiritin-7-O-glucoside-4’-O-glucosyl-apioside,vicenin-2,schaftoside,isoviolanthin,chrysin-6-C-glucosyl-8-C-arabonoside,chrysin-6-C-arabinosyl-8-C-glucoside,liquiritin apioside,isoacteoside,liquiritin,scutellarin,acteoside,5,6’-dihydroxy-7,8-dimethoxy-2f-O-β-D-glucopyranosylflavone,chrysin-8-C-β-D-glucopyranoside,5,2’,6’-trihydroxy-6,7,8-trimethoxyflavone-2’-O-glucoside,licuraside,cistanoside D,5,8-dihydroxy-6-methoxy-7-O-glu acid,isoliquiritin,cosmosiin,neoisoliquiritin,baicalin,5,6-digydroxy-7-glucuronic acid flavanone,galengin 7-O-glucuronide,5,2’-dihydroxy-6-methoxy-7-O-glu acid,oroxylinA-7-O-β-D-glu acid,chrysin-7-O-β-D-glucuronopyranoside,5,6-dihydroxy-6-methoxy-7-O-glu acid,wogonoside,5,7-dihydroxy-6-methoxyflavone-7-glucurronide methylester),柴胡皂苷(acetyl-saikosaponin A or D,SSA,saikosaponin SSb1 or SSb2,acetyl-saikosaponin A or D,diacetyl-Saikosaponin A or D,acetyl-Saikosaponin A or D,SSD),甘草皂苷(licoricesaponin E2,22β-acetoxylglycyrrhizic acid,yungaonoside K2,licoricesaponin G2,glycyrrhizin acid or licoricesaponin-H2 or licoricesaponin-K2,licoricesaponin-J2,licoricesaponin B2,glycyrrhizin acid or licoricesaponin-H2 or licoricesaponin-K2,glycyrrhizin acid or licoricesaponin-H2 or licoricesaponin-K2,licoricesaponin A3),人参皂苷(ginsenoside Rg1,ginsenoside Rf,isomer or ginsenoside Rg2,isomer or ginsenoside Rg2,ginsenoside Rb1,isomer or ginsenoside Rg6 or F4,isomer or ginsenoside Rg6 or F4,isomer or ginsenoside Rg6 or F4,ginsenoside Re)、和生姜素(6-gingerol,methy[8]-gingerol,[6]-shogaol,isomer or[6]-paradol)等在内的79个化学成分,最终确定小柴胡汤全方69个化合物的化学结构。通过活性追踪引导成分分离结合LC-ESI-QTOF-MS/MS的初步研究结果,推测多糖,生姜和黄酮类化合物可能对小柴胡汤抗抑郁作用有较大的贡献。综上所述,活性追踪引导成分分离结合LC-ESI-QTOFMS/MS的方法对初步鉴别复杂的中药复方成分有很好应用。半夏是小柴胡汤中的组方药材,半夏化学成分的研究是小柴胡汤拆方化学成分研究的一部分。半夏来源于天南星科(Araceae)半夏属植物半夏Pinellia ternata(Thunb.)Breit的干燥块茎。利用多种色谱分离技术,从半夏块茎(15 kg)的75%乙醇提取物中分离得到45个单体化合物,运用1D,2D NMR analyses,HR-ESI-MS,CD等波谱技术确定了其中40个化合物的结构,其中新化合物1个已知化合物39个分别为:cyclolignanyingoside A(41),isolariciresinol 9-O-β-D-glucopyranoside(42),isolariciresinol(43),verticillatoside A(44),pinoresinol-4’-O-β-D-glucoside(45),medioresinol(46),Pinoresinol(47),burselignan(48),dehydrodiconiferyl alcohol-9-O-β-D-glucopyranoside(49),dehydrodiconiferylalcohol-9’-O-β-D-glucopyranoside(50),dehydrodiconiferylalcohol-4,9-di-O-β-D-glucoside(51),lariciresinol 4-O-β-D-glucopyranoside(52),(7R,8R)-1-(4-hydroxy-3-methoxyphenyl)-2-{4-[(E)-3-hydroxyl-l-propenyl]-2-methoxyphe-noxy}-1,3-propanediol(53),cinnacassoside A(54),neo-olivil(55),americanol A(56),isoamericanol A(57),forsythiayanoside B(58),matairesinoside(59),syringaresinol-4-O-β-D-glucopyrranoside(60),cyclo-Prolyl-Lecine(61),cyclo-Pro-Ile(62),cyclo-Pro-Val(63),uridine(64),adenosine threo-2S-3-O-methyl-guaiacylglycerol(65),erythro-2-R-3-O-methyl-guaiacylglycerol(66),3,4-dihydroxybenzoic acid(67),2,4-dimethoxyphenyl-1-O-β-D-glucopyranoside(68),coniferaldehyde(69),vanillic(70),4-O-β-D-glucopyranosylvanilylalcohol(71),3,4-dimethoxybenzyl-β-D-glucopyranoside(72),3,4-dimethoxybenzyl-β-D-glucopyranoside(73),p-hydroxybenzoic acid(74),4-(2-hydroxyethoxy)-3-methoxy-benzoic acid(75),sachalide 1(76),nikoenoside(77),3-methoxy-4-methylbenzoic acid(78),p-methoxy phenylpropyl aldehyde(79),trans-p-hydroxy cinamic acid(80).对半夏中分离得到的7个木脂素类化合物进行了 3种人肿瘤细胞株(Hep-3B,Bcap-37 and MCF-7)的体外细胞毒活性和抗氧化活性(DPPH和ABTS)考察。结果显示:化合物对3种人肿瘤细胞株均无显著的抗肿瘤活性,化合物1-7均有不同程度的抗氧化活性。此外,本实验还对半夏中分离得到的15个木脂素类成分的可能生物合成途径进行了推测,并运用多反应离子监测技术(MRM)建立了同时测定多批次半夏及其伪品(水半夏和天南星)中10种木脂素类化合物的含量测定方法。结果表明化合物5、6和10可作为区分半夏及其伪品(水半夏和天南星)的潜在化合物。