三线定量胶体金免疫亲和试纸法定量中药饮片中黄曲霉毒素B1及B1、B2、G1、G2总量的研究  

Quantitative study of total amount of aflatoxins B1 and B1,B2,G1 and G2 in Chinese herbal pieces by colloidal gold immunoaffinity method

作　　者：范妙璇[1] 傅欣彤[1] 陈奕菲 陈思妮 陈晶[1] 张婷 陈有根[1] FAN Miao-xuan;FU Xin-tong;CHEN Yi-fei;CHEN Si-ni;CHEN Jing;ZHANG Ting;CHEN You-gen(Beijing Key Laboratory of Analysis and Evaluation on Chinese Medicine,NMPA Key Laboratory for Quality Evaluation of Traditional Chinese Patent Medicine,Beijing Institute for Drug Control,Beijing 102206,China)

机构地区：[1]北京市药品检验所,国家药品监督管理局中成药质量评价重点实验室,中药成分分析与生物评价北京市重点实验室,北京102206

出　　处：《中草药》2021年第17期5275-5286,共12页Chinese Traditional and Herbal Drugs

摘　　要：目的利用三线胶体金侧流向免疫色谱技术,针对中药特点对样品前处理和检测曲线进行研究,建立快速、准确的定量测定中药饮片中黄曲霉毒素B1、B2、G1和G2总量的方法,迅速检测中药饮片中黄曲霉毒素的污染程度。方法利用胶体金免疫亲和法建立《中国药典》2020年版规定限度的24种中药饮片中黄曲霉毒素测定方法,进行方法学验证,并对60种中药饮片,共计195批次样品进行分类测定。同时利用液质联用(三重四级杆质谱法)对上述样品进行定量检测,并比对2种检测方法。采用配对t检验法比较2种方法的检测结果是否存在显著性差异;通过加入其他真菌毒素对试纸条的特异性进行考察。结果通过胶体金侧流向免疫色谱技术测定中药饮片中黄曲霉毒素含量,胶体金方法和液质联用方法检测结果一致,无显著性差异,加样回收率为80.3%~119.7%,与黄曲霉毒素M1和M2、赭曲霉毒素、脱氧雪腐镰刀菌烯醇和玉米赤霉烯酮等其他真菌毒素没有交叉反应。单独研究黄曲霉毒素B1与B1、B2、G1和G2总量之间无交叉干扰,且其含量测定符合《中国药典》2020年版规定的标准检验要求。结论胶体金检测方法可以准确、定量、快速检测中药饮片中黄曲霉毒素含量。该方法具有简单、快捷和低毒等优点。Objective A rapid and accurate method for quantitative determination of aflatoxins B1,B2,G1,and G2 in Chinese herbal pieces was established by using colloidal gold-side flow immunochromatography,and the pollution level of aflatoxins in prepared slices of Chinese herbal pieces was determined rapidly.Methods The content of aflatoxin in 24 kinds of Chinese herbal pieces was determined by colloidal gold immunoaffinity method,and the results were compared by liquid mass spectrometry.The paired t test was used to compare whether there was significant difference between the two methods.The specificity of the test strip was investigated by adding other fungal toxins.Tonga calculated the recovery rate.Results The aflatoxin content in the Chinese herbal pieces were determined through side to immune colloidal gold chromatography technology.The test results of colloidal gold method and LCMS/MS method were consistent.There was no significant difference,result in three levels in the low recovery rate was between 80.3%and 119.7%,and aflatoxin M1 and M2,ochre and aspergillus toxin,vomiting toxins and corn gibberellic ketene and other mycotoxins had no cross reaction,which met the requirements.Conclusion Colloidal gold assay can be used for accurate,quantitative and rapid determination of aflatoxins in prepared slices.The method has the advantages of simplicity,rapidity and low toxicity.

关 键 词：黄曲霉毒素 三线胶体金侧流向免疫色谱技术 胶体金免疫亲和法 中药饮片 液质联用法 赭曲霉毒素 脱氧雪腐镰刀菌烯醇 玉米赤霉烯酮 

分 类 号：R283.6[医药卫生—中药学]