基于简单样品前处理的玉米赤霉烯酮酶联免疫分析方法——以瓜蒌皮和瓜蒌子的检测为例  

Development of an enzyme-linked immunoassay for zearalenone based on simple pretreatment technology — A case study of Trichosanthis pericarpium and Trichosanthis semen detection

作　　者：张磊 吴旺荣 黄仁堂 关凯仪 王淑美 ZHANG Lei;WU Wangrong;HUANG Rentang;GUAN Kaiyi;WANG Shumei(School of Chinese Materia Medica,Guangdong Pharmaceutical University,Guangzhou 510006,China;Key Laboratory of Digital Quality Evaluation of Traditional Chinese Medicine,National Administration of Traditional Chinese Medicine,Guangzhou 510006,China;Guangdong Provincial Traditional Chinese Medicine Quality Engineering and Technology Research Center,Guangzhou 510006,China;School of Pharmacy,Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]广东药科大学中药学院,广东广州510006 [2]国家中医药管理局中药数字化质量评价技术重点研究室,广东广州510006 [3]广东省中药质量工程技术研究中心,广东广州510006 [4]广东药科大学药学院,广东广州510006

出　　处：《广东药科大学学报》2023年第2期32-36,共5页Journal of Guangdong Pharmaceutical University

基　　金：国家自然科学基金项目(81903798);广东省基础与应用基础研究基金(2022A1515012099)。

摘　　要：目的以瓜蒌皮和瓜蒌子中玉米赤霉烯酮的检测为例,探讨简便、高效的样品前处理技术,解决酶联免疫分析法检测中药中真菌毒素的基质干扰问题。方法首先优化抗原-抗体最佳反应浓度,建立间接竞争酶联免疫分析抑制曲线,确定灵敏度和线性范围;再系统对比不同提取溶剂及稀释溶液获取最佳样品前处理过程;经方法学考察后,将所建立的方法用于实际样品测定。结果瓜蒌皮和瓜蒌子的检测限分别为0.017 ng/mL和0.031 ng/mL,线性范围分别为0.0625~2 ng/mL和0.094~2 ng/mL,加标回收率为75.88%~104.7%和78.03%~104.7%(RSD<8%);该方法成功用于11批瓜蒌皮和17批瓜蒌子样品检测,未检出阳性样品。结论适宜的提取溶剂和稀释溶液组合可有效解决基质干扰问题,该方法操作简便,准确性好,为其他中药中真菌毒素快速筛查方法的建立提供参考。Objective Taking the zearalenone detection in Trichosanthis pericarpium and Trichosanthis semen as an example,a simple and efficient sample pretreatment technology was discussed to solve the matrix interference problem of enzyme-linked immunosorbent assay in the detection of mycotoxins in traditional Chinese medicine.Methods Firstly,the optimal concentration of antigen-antibody reaction was optimized,and the indirect competitive enzyme-linked immunoassay inhibition curve was established to determine the sensitivity and linear range.Then the optimal sample pretreatment process was obtained by systematically comparing different extraction solvents and diluted solutions to reduce matrix interference.After method validation,the established method was applied to the real sample analysis.Results The detection limits of Trichosanthis pericarpium and Trichosanthis semen were 0.017 ng/mL and 0.031 ng/mL,respectively,with a linear range of 0.0625-2 ng/mL and0.094-2 ng/mL,respectively.The recoveries were 75.88%-104.7% and 78.03%-104.7%,respectively,with RSDs less than 8%.This method was used to detect 11 batches of Trichosanthis pericarpium samples and 17 batches of Trichosanthis semen samples,and no positive samples were found.Conclusion The results show that the suitable combination of extraction solvent and dilution solution can effectively overcome the matrix interference.The method is simple and accurate,which can provide a reference for establishing rapid screening methods for mycotoxins in other traditional Chinese medicines.

关 键 词：玉米赤霉烯酮 酶联免疫分析 样品前处理 快速检测 基质效应 

分 类 号：R284.1[医药卫生—中药学]