高效液相色谱-电喷雾离子阱质谱法鉴定金匮肾气丸总苷化学成分及代谢产物

Identification of Chemical and Metabolic Components of Total Glycosides of Jinkuishenqi Pill by High Performance Liquid Chromatography-Electrospray Ion Trap Mass Spectrometry

应用高效液相色谱-电喷雾离子阱质谱法(HPLC-ESI/MSn)鉴定金匮肾气丸总苷的化学成分及总苷大鼠灌胃给药血浆化学成分或代谢产物。对比体内外样品色谱图各色谱峰,根据负离子模式下的分子离子峰获得化合物分子量信息,金匮肾气丸总苷含19个化学成分:没食子酰-3-O-芹糖基(1→6)葡糖苷、羟基芍药苷、莫诺苷、马钱苷、獐芽菜苷、地黄苷A或B、1,2,3-三-O-没食子酰葡萄糖、芍药苷、山茱萸新苷Ⅱ、6'-(3,4,5-三羟基苯甲酰)芍药苷、1,2,3,6-四-O-没食子酰葡萄糖、山茱萸新苷Ⅰ、五没食子酰葡萄糖、苯甲酰羟基芍药苷、丹皮酚原苷、苯甲酰芍药苷、4'-羟基,6'-(3,4,5-三羟基苯甲酰)芍药苷和两个未知化合物成分。金匮肾气丸总苷大鼠灌胃血浆鉴定出17个化学成分,其中有8个化合物以原型存在,9个可知代谢转化成分,分别为没食子酸,2-羟基苯乙酮-4-O-葡萄糖醛酸酯,芍药苷代谢素Ⅰ,2-羟基-4-甲氧基苯乙酮-5-O-硫酸酯,2,4-二羟基苯乙酮-5-O-硫酸酯,2,4-二羟基苯乙酮和3个未知成分。

To identify the chemical components of total glycosides of Jinkuishenqi pill and their metabolites in vivo,analysis and comparison were carried out by High performance liquid chromatography electrospray ion trap mass spectrometry(HPLC-ESI-MS) among rats' plasma samples obtained after oral administration of total glycoside(TG) of Jinkuishenqi pill.19 compounds,including galloyl-3-O-apiofuranosyl-(1→6)-glucoside,hydroxyl-paeoiflorin,morroniside,loganin,sweroside,rehmaionoside A or B,1,2,3-tri-O-galloyl-β-D-glucose,paeoniflorin,cornuside II,6'-O-(3,4,5-trihydroxybenzoyl) paeoniflorin,1,2,3,6-tetra-O-galloyl-βD-glucose,cornuside I,pentagalloylglucose,hydroxy-benzoyl paeoniflorin,paeonoside,benzoylpaeoniflorin,4'-hydroxy,6'-(3,4,5-trihydroxy-benzoyl)-paeoniflorin and 2 compounds unknown,were detected in TG of Jinkuishenqi pill.The compounds absorbed into blood,including their prototype components and their metabolites,were identified.17 compounds were detected after oral administration of TG,9 of which are metabolites and others are the original form of the compounds contained in Jinkuishenqi pill.Among the total,the metabolites are Galic acid,2-hydroxyacetophenone-4-O-glucuronic acid esters,paeonimetabolin Ⅰ,2-hydroxy-4methoxy acetophenone-5-O-sulfate,2,4-dihydroxyacetophenone-5-O-sulfate,2,4-dihydroxyacetophenone and 3 compounds unknown,respectively.The results showed that HPLC-ESI/MS method is efficient,sensitive for detection and identification of prototype and metabolism of total glucosides of metabolites in vivo,which is important to reveal the chemical component of the metabolism in vivo.