基于序贯代谢方法的痛风定胶囊体内代谢分析  

Metabolic Analysis of Tongfengding Capsules in vivo Based on Sequential Metabolism

作　　者：陈洪娇 杨文宁[1] 尹仁芳[2] 朱彦[2] 祁东盈 李雪岩 刘慧宁 于爽 潘艳丽[2] CHEN Hongjiao;YANG Wenning;YIN Renfang;ZHU Yan;QI Dongying;LI Xueyan;LIU Huining;YU Shuang;PAN Yanli(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;Institute of Information on Traditional Chinese Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]北京中医药大学中药学院,北京102488 [2]中国中医科学院中医药信息研究所,北京100700

出　　处：《中国实验方剂学杂志》2022年第24期160-170,共11页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81973295)。

摘　　要：目的:采用序贯代谢方法,结合超高效液相色谱-高分辨质谱法(UPLC-HRMS),鉴定痛风定胶囊在大鼠体内的原型成分和代谢物。方法:采用ACQUITY UPLC BEH Shield RP18色谱柱(2.1 mm×100 mm,1.7μm),以0.1%甲酸水溶液(A)-乙腈(B)为流动相进行梯度洗脱(0~1 min,5%B;1~2.4 min,5%~10%B;2.4~13.5 min,10%~32%B;13.5~18.5 min,32%~90%B;18.5~19 min,90%~5%B;19~21 min,5%B),流速0.3 mL·min^(-1),进样量2μL,柱温35℃,运用加热电喷雾离子源(HESI),扫描范围m/z 100~1 500,正、负离子模式检测。通过比较给药血浆和空白血浆的差异,对由肠道灌流并行采血制备得到的肠代谢、肝代谢血浆样品及灌胃给药方法制备得到的综合代谢血浆样品中的原型入血成分和代谢产物进行鉴定。结果:从肠代谢、肝代谢及综合代谢样品中分别检测到了76、53、74个化学成分,从上述不同血浆样品中共鉴定出100个化学成分,包括64种原型入血成分(34个生物碱类、12个萜类、9个有机酸类、6个黄酮类与3个其他类成分)与36种代谢产物,代谢产物生成过程中涉及的代谢反应主要有葡萄糖醛酸化、去葡萄糖基化、脱氢化与羟基化。结论:痛风定胶囊中化学成分在肠道与肝脏处会发生一系列代谢反应,生成大量代谢产物,其中生物碱类可能是其主要成分群,可为该制剂体内药效物质基础的系统阐释奠定基础。Objective:Based on sequential metabolism and ultra-high performance liquid chromatography-high resolution mass spectrometry(UPLC-HRMS),to identify the prototype components and metabolites of Tongfengding capsules in rat plasma.Method:An ACQUITY UPLC BEH Shield RP18 column(2.1 mm×100 mm,1.7μm)was used for gradient elution with 0.1% formic acid aqueous solution(A)-acetonitrile(B)as the mobile phase(0-1 min,5%B;1-2.4 min,5%-10%B;2.4-13.5 min,10%-32%B;13.5-18.5 min,32%-90%B;18.5-19 min,90%-5%B;19-21 min,5%B)at a flow rate of 0.3 mL·min,injection volume of 2μL and column temperature at 35℃.The heated electrospray ionization(HESI)was applied under positive and negative ion modes,and the scanning range was m/z 100-1500.By comparing the differences between the administered plasma and the blank plasma,the prototype components and metabolites in intestina metabolism sample and liver metabolism sample prepared by intestinal perfusion with parallel blood collection,and comprehensive metabolism sample prepared by intragastric administration method were identified.Result:A total of 76,53,74 chemical components were detected in the intestinal metabolism sample,the liver metabolism sample and the comprehensive metabolism sample.A total of 100 components were identified from these different plasma samples,including 64 prototype components(34 alkaloids,12 terpenoids,9 organic acids,6 flavonoids and 3 other components)and 36 metabolites.The main metabolic reactions involved in the formation of metabolites were glucuronidation,deglucosylation,dehydrogenation and hydroxylation.Conclusion:The chemical components of Tongfengding capsules can undergo a series of metabolic reactions in the intestine and liver,and a large number of metabolites are generated,among which alkaloids may be the leading component group for efficacy,which can lay the foundation for systematic elucidating the material basis of Tongfengding capsules in vivo.

关 键 词：痛风定胶囊 序贯代谢方法 超高效液相色谱-高分辨质谱法(UPLC-HRMS) 羟基化 葡萄糖醛酸化 生物碱 萜类 

分 类 号：R22[医药卫生—中医基础理论] R28[医药卫生—中医学] R96[理学—分析化学] O657[理学—化学]