基于UHPLC-MS/MS的钩藤生物碱在大鼠体内的组织分布研究  

Tissues distribution of alkaloids from Uncaria rhynchophylla in rats by UHPLC-MS/MS

作　　者：吴依松 李士伟 李昊举 魏文龙 吴士飞 果德安 WU Yi-song;LI Shi-wei;LI Hao-ju;WEI Wen-long;WU Shi-fei;GUO De-an(College of Pharmacy,Changchun University of Chinese Medicine,Changchun 130117,China;Shanghai Research Center for Modernization of Traditional Chinese Medicine,National Engineering Research Center for TCM Standardization Technology,Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Shanghai 201203,China)

机构地区：[1]长春中医药大学药学院,吉林长春130117 [2]中国科学院上海药物研究所上海中药现代化研究中心,中药标准化技术国家工程研究中心,上海201203

出　　处：《中草药》2022年第13期4035-4045,共11页Chinese Traditional and Herbal Drugs

基　　金：上海市青年科技英才扬帆计划资助项目(21YF1455800)。

摘　　要：目的建立超高效液相串联质谱(ultra-high performance liquid chromatography-tandem mass spectrometry,UHPLCMS/MS)法测定大鼠不同组织中钩藤碱、异钩藤碱、去氢钩藤碱、异去氢钩藤碱、毛钩藤碱、去氢毛钩藤碱与缝籽嗪甲醚的含量,并用于钩藤的组织分布研究。方法大鼠ig钩藤水煎液后,分别于0.083、0.25、0.5、0.75、1、2、4、8、12 h取心、肝、脾、肺、肾、脑组织。以长春胺为内标,采用液液萃取法提取后进行样本检测。结果建立的UHPLC-MS/MS方法中,7个生物碱在不同的组织中最低定量限范围为0.05~1.00 ng,响应与浓度的相关性R≥0.991。方法学考察中的专属性、基质效应、回收率、精密度与准确度以及稳定性等方法学考察均满足要求。各个组织中均为异钩藤碱含量最高,其次为去氢钩藤碱;异钩藤碱与去氢钩藤碱含量均远高于其对应的异构体钩藤碱与异去氢钩藤碱;而3个四环单萜吲哚生物碱(毛钩藤碱、去氢毛钩藤碱、缝籽嗪甲醚)在各组织中均含量最低。比较不同组织中生物碱成分含量可以发现,7个生物碱均在肝脏中含量最高,其次为肾脏与肺部,推测钩藤生物碱在肝脏与肾脏的蓄积是因为两者分别是生物碱成分的主要代谢部位与排泄部位。结论建立的方法灵敏度高、准确性好、专属性好,适用于大鼠不同组织中钩藤生物碱的组织分布研究。Objective To establish an ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)method for determination of contents of rhynchophylline(RHY),isorhynchophylline(IRHY),corynoxeine(COE),isocorynoxeine(ICOE),hirsutine(HTI),hirsuteine(HTE)and geissoschizine methyl ether(GSM)and applied for tissue distribution of Uncaria rhynchophylla.Methods Rats were ig U.rhynchophylla decoction,then heart,liver,spleen,lung,kidney and brain tissue were acquired at 0.083,0.25,0.5,0.75,1,2,4,8,12 h.Vincamine was used as internal standard,samples were extracted by liquid-liquid extraction.Results For the established UHPLC-MS/MS method,minimum quantitative limits of seven alkaloids in different tissues ranged from 0.05 to 1.00 ng,and correlation between response and concentration was R~2≥0.991.The specificity,matrix effect,recovery rate,precision and accuracy and stability met the requirements.IRHY content was the highest in each tissue,followed by COE;Both IRHY and COE were much higher than their corresponding isomers(RHY and ICOE),while three tetracyclic monoterpene indole alkaloids(HTE,HTI and GSM)were the lowest in all tissues.Comparing the contents of alkaloids in different tissues,it was found that content of seven alkaloids was the highest in liver,followed by kidney and lung.The cause of accumulation was probably due to kidney and liver to be the main sites of metabolism and excretion.Conclusion The established method has high sensitivity,good accuracy and specificity,which was suitable for tissue distribution study of alkaloids from U.rhynchophylla in different tissues of rats.

关 键 词：钩藤 生物碱 钩藤碱 异钩藤碱 去氢钩藤碱 异去氢钩藤碱 毛钩藤碱 去氢毛钩藤碱 缝籽嗪甲醚 定量分析 组织分布 

分 类 号：R285.5[医药卫生—中药学]