基于UHPLC-Q-Exactive Orbitrap MS鉴定参芪降糖胶囊在大鼠体内的代谢产物

Identification of metabolites of Shenqi Jiangtang Capsules in rats based on UHPLC-Q-Exactive Orbitrap MS

目的研究参芪降糖胶囊在大鼠血浆、胆汁、尿液和粪便中的代谢产物及其代谢途径。方法采用UHPLC-Q-Exactive OrbitrapMS技术,以0.1%甲酸溶液(A)-乙腈(B)为流动相进行梯度洗脱,并采用ESI离子源,分别在正、负离子模式进行全扫描和二级质谱扫描,获得化合物的一级和二级质谱数据,结合文献报道、对照品裂解规律及药物代谢反应规律,对大鼠ig参芪降糖胶囊混悬液后的生物样品进行分析鉴定。结果在大鼠体内共鉴定出125个化合物,包括47个原型成分(PA1~PF)和78个代谢产物(MA1~ME14),其中2个为新代谢产物,分别为MC15(五味子甲素双脱甲基后葡萄糖醛酸化产物)和MD3(去氢茯苓酸的硫酸酯化产物),4个为新化合物,分别为MC14:(6R,7R)-2,3,10,11-tetramethoxy-6,7-dimethyl-5,6,7,8-tetrahydrodibenzo[a,c][8]annulene-1,7-diol、MC16:(6R,7R)-2,3,10-trimethoxy-6,7-dimethyl-5,6,7,8-tetrahydrodibenzo[a,c][8]annulene-1,7,11-triol、MD1:(R)-2-((3R,3a R,6S,7S,9b R)-6-(2-carboxyethyl)-3a,6,9b-trimethyl-7-(prop-1-en-2-yl)-3a,4,6,7,8,9bhexahydro-3H-cyclopenta[a]naphthalen-3-yl)-6-methylhept-5-enoicacid和MD4:(3S,5R,10S,13R,14R,16R,17R)-4,4,10,13,14,17-hexamethyl-2,3,4,5,6,10,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthrene-3,16-diol。结论参芪降糖胶囊在大鼠体内的代谢途径主要涉及脱羟基、脱甲基、脱水、水解、还原氢化、甲基化、葡萄糖醛酸化、硫酸酯化和多种反应类型的复合反应等。初步阐明了参芪降糖胶囊的体内代谢特征,同时为探索其生物活性成分及作用机制提供参考。

Objective To investigate the metabolites and their major metabolic pathways of Shenqi Jiangtang Capsules(SJC,参芪降糖胶囊)in rat plasma,bile,urine and feces biological samples.Methods Based on UHPLC-Q-Exactive Orbitrap MS technology,gradient elution was carried out with 0.1%formic acid solution(A)-acetonitrile(B)as the mobile phase,and full scanning and secondary mass spectrometry scanning were carried out with ESI ion source in positive and negative ion modes,respectively,to obtain the primary and secondary mass spectrometry data of the compounds.Combined with literature reports,the fragmentation pattern of reference substance and the law of drug metabolism reaction,the biological samples of rats after ig administration of SJC suspension were analyzed and identified.Results A total of 125 compounds were identified in rats,including 47 prototype components(PA1—PF)and 78 metabolites(MA1—ME14),of which two were novel metabolites,namely MC15(glucuronic acid product after di-demethylation of schizandrin A)and MD3(sulfuric acid esterification product of dehydropachymic acid),and four were new compounds,namely(6R,7R)-2,3,10,11-tetramethoxy-6,7-dimethyl-5,6,7,8-tetrahydrodibenzo[a,c][8]annulene-1,7-diol(MC14),(6R,7R)-2,3,10-trimethoxy-6,7-dimethyl-5,6,7,8-tetrahydrodibenzo[a,c][8]annulene-1,7,11-triol(MC16),(R)-2-((3R,3aR,6S,7S,9bR)-6-(2-carboxyethyl)-3a,6,9b-trimethyl-7-(prop-1-en-2-yl)-3a,4,6,7,8,9b-hexa hydro-3H-cyclopenta[a]naphthalen-3-yl)-6-methylhept-5-enoic acid(MD1)and(3S,5R,10S,13R,14R,16R,17R)-4,4,10,13,14,17-hexamethyl-2,3,4,5,6,10,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthrene-3,16-diol(MD4).Conclusion Research revealed that dehydroxylation,demethylation,dehydration,hydrolysis,reductive hydrogenation,methylation,glucuronidation and sulfation were the main metabolic pathways of SJC in rat samples.The preliminarily clarified in vivo metabolic characteristics of SJC provided a reference for exploring bioactive ingredients and action mechanisms of SJC.