免疫亲和柱-高效液相色谱法测定炒僵蚕中黄曲霉毒素

Detection of Aflatoxin in Fried Bombyx Batryticatus by Immunoaffinity Chromatography-High Performance Liquid Chromatography

目的:采用免疫亲和柱-高效液相色谱法,对炒僵蚕饮片中的黄曲霉毒素B_(1)、B_(2)、G_(1)、G_(2)的残留量进行分析,旨在为炒僵蚕饮片质量标准的完善及其市场监管提供依据。方法:样品经过高速匀浆处理,经免疫亲和柱净化,采用高效液相色谱-柱后光化学衍生法测定。采用YMC-C_(18)(250 mm×4.6 mm,5μm)色谱柱,以甲醇-乙腈-水(32∶18∶50)为流动相,流速为1.0 mL·min^(-1),柱温为30℃,荧光检测器激发波长为360 nm,发射波长为450 nm,对7批炒僵蚕进行分析。结果:本研究采用的黄曲霉毒素B_(1)、B_(2)、G_(1)、G_(2)检测方法的线性关系、精密度、回收率良好,黄曲霉霉毒素G_(2)在多批样品检出,检出率为86%,有1批样品中黄曲霉霉毒素G_(1)含量较高,参照《中华人民共和国药典》(2020年版)黄曲霉毒素的限量标准,其黄曲霉毒素B_(2)、G_(1)和G_(2)总量超出限度规定。结论:本研究为首次采用免疫亲和柱-高效液相色谱法对炒僵蚕中黄曲霉素进行筛查,明确其存在安全风险,为炒僵蚕饮片中黄曲霉素的质控提供参考。

Objective:Immunoaffinity chromatography-high performance liquid chromatography(IAC-HPLC)was used to detect the aflatoxin B_(1),B_(2),G_(1) and G_(2) in fired Bombyx batryticatus,so as to provide reference for the improvement of the quality standard of fired Bombyx batryticatus and its market supervision.Methods:The analysis of 7 batches of fried Bombyx batryticatus was performed on a YMC-C_(18)(250 mm×4.6 mm,5μm)column with methanol-acetonitrile-water(32∶18∶50)as the mobile phase at a flow rate of 1.0 mL·min^(-1).The column temperature was 30℃.The excitation and emission wavelengths of the fluorescence detector were 360 nm and 450 nm,respectively.Results:The detection methods of aflatoxin B_(1),B_(2),G_(1) and G_(2) adopted in this study showed good linearity,precision and recovery.Aflatoxin G_(2) was detected in multiple batches of samples,with a detection rate of 86%.The content of aflatoxin G_(1) in one batch of samples was higher.According to the limit standard of aflatoxin in Chinese Pharmacopoeia 2020 edition,the total amount of aflatoxin B_(2),G_(1) and G_(2) exceeded the limit.Conclusion:This study is the first time to detect aflatoxin in fried Bombyx Batrvticatus bimmunoaffinity column-high performance liquid chromatography,and to clarify the safety risks otaflatoxin in fried Bombyx Batryticatus,so as to provide reference for the quality control of aflatoxirin fried Bombyx Batryticatus.