基于酶联免疫分析法高通量筛查中药材中赭曲霉毒素A的污染研究  

High-throughput screening of ochratoxin A in Chinese herbal medicines using enzyme-linked immunoassay

作　　者：秦露 张磊 蒋佳伊 王长健 豆小文[2] 万丽[1] 杨美华[2] QIN Lu;ZHANG Lei;JIANG Jia-yi;WANG Chang-jian;DOU Xiao-wen;WAN Li;YANG Mei-hua(College of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine,Ministry of Education,Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100193,China)

机构地区：[1]成都中医药大学药学院,四川成都611137 [2]中国医学科学院北京协和医学院药用植物研究所中草药物质基础与资源利用教育部重点实验室,北京100193

出　　处：《中国中药杂志》2019年第23期5072-5077,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81872999,81703699);中国医学科学院医学与健康科技创新工程项目(2017-I2M-1-013);国家重点研发计划项目(2016YFE0112900);国家级外专项目(G20190001631)

摘　　要：建立了一种间接竞争酶联免疫吸附法(ic-ELISA)快速检测3种中药材肉豆蔻、干姜和姜黄中赭曲霉毒素A(OTA)的方法,采用基质匹配标准曲线代替样品稀释液标准曲线,并对样品提取液和样品稀释液进行优化。该方法检测肉豆蔻、干姜和姜黄中OTA的灵敏度(IC50)分别为0. 146,0. 157,0. 153 ng·m L-1,检出限(LOD)分别为0. 040,0. 032,0. 031 ng·m L-1,样品回收率为75. 99%~122. 3%,RSD<10%。50份样品中检测到2份肉豆蔻阳性样品和1份姜黄阳性样品,OTA检出最高值为1 167. 8μg·kg^-1。样品检测结果采用LC-MS/MS进行确证。研究中建立的ic-ELISA方法简单、快速、灵敏,可用于中药肉豆蔻、干姜和姜黄及其他中药中OTA快速筛查。An indirect competitive enzyme-linked immunosorbent assay( ic-ELISA) was developed for the rapid detection of ochratoxin A( OTA) in nutmeg( Myristicae Semen),ginger( Zingiberis Rhizoma) and turmeric( Curcumae Longae Rhizoma). The matrix matching standard curve was used instead of the standard curve of sample diluent,and the sample extract and sample diluent were optimized. The sensitivity( IC50) of this method for OTA in nutmeg,ginger and turmeric were determined as 0. 146,0. 157 and 0. 153 ng·m L-1,respectively and the limits of detection( LODs) were 0. 040,0. 032 and 0. 031 ng·m L-1,respectively. The recovery of samples ranged from 75. 99% to 122. 3%,with RSD<10%. Two positive samples for nutmeg and one positive sample for turmeric occurred in 50 samples,and the highest OTA contamination value was 1 167. 8 μg·kg^-1. The results were further confirmed by LC-MS/MS. It shows that the developed ic-ELISA method is simple,rapid and sensitive,and can be applied for rapid and high-throughput screening of OTA in nutmeg,ginger and turmeric,as well as some other CHMs.

关 键 词：间接竞争酶联免疫吸附法 肉豆蔻 干姜 姜黄 赭曲霉毒素A 快速筛查 

分 类 号：R284.1[医药卫生—中药学]