免疫磁珠高通量全自动净化结合高效液相色谱快速测定饲料中玉米赤霉烯酮含量  

Determination of Feed Zearalenone Content Using High-Throughput Automatic Immunomagnetic Beads Purification Combined with High-Performance Liquid Chromatography

作　　者：莫凡 姚婷 胡深 索德成[1] 赵新雪 邹宇飞 王紫豪 肖志明[1] MO Fan;YAO Ting;HU Shen;SUO Decheng;ZHAO Xinxue;ZOU Yufei;WANG Zihao;XIAO Zhiming(Institute of Quality Standard and Testing Technology for Agro-Products,Chinese Academy of Agricultural Sciences,Beijing 100081,China;Beijing Veterinary Drug and Feed Monitoring Center,Beijing 102200,China;Hubei Province Feed Monitoring Institute,Wuhan 430048,China;Suzhou Beaver Biomedical Engineering Co.,Ltd.,Suzhou 215123,China;Hebei Elisa Biotechnology Co.,Ltd.,Baoding 072750,China)

机构地区：[1]中国农业科学院农业质量标准与检测技术研究所,北京100081 [2]北京市兽药饲料监测中心,北京102200 [3]湖北省饲料监测所,武汉430048 [4]苏州海狸生物医学工程有限公司,苏州215123 [5]河北伊莱莎生物技术有限公司,保定072750

出　　处：《动物营养学报》2024年第11期7454-7466,共13页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家重点研发计划(2023YFD1301001);中国农业科学院基本科研业务费(1610072023005,1610072024009);中国饲料工业协会团体标准制定项目。

摘　　要：本研究旨在建立利用免疫磁珠高通量全自动净化结合高效液相色谱快速测定饲料中玉米赤霉烯酮含量的方法。饲料样品用80%乙腈水提取,经全自动免疫磁珠净化、富集后,采用高效液相色谱-荧光检测器(HPLC-FLD)测定,外标法定量。结果表明:玉米赤霉烯酮在0.01~1.00μg/mL范围内线性关系良好(相关系数≥0.9999),方法检出限为2μg/kg,定量限为10μg/kg;玉米赤霉烯酮在0.01、0.02、0.10和1.00 mg/kg添加水平下的平均回收率为86.90%~100.27%,批内相对标准偏差(RSD)为3.18%~9.88%,批间RSD为4.94%~9.40%。由此可见,与传统固相萃取、免疫亲和柱净化法相比,免疫磁珠净化集提取、分离、净化、浓缩于一体,可实现完全自动化处理,提高工作效率和检测通量,避免因人为操作导致的结果偏差,适用于大批量饲料样本中玉米赤霉烯酮含量的测定。This experiment was to study the method for determination of feed zearalenone content using high-throughput automatic immunomagnetic beads purification combined with high-performance liquid chromatography.Feed samples were extracted with 80%acetonitrile water,purified and enriched by automatic immunomagnetic beads,and determined by high-performance liquid chromatography-fluorescence detector(HPLC-FLD),with external standard method for quantification.The results indicated that good linearity was obtained for zearalenone in the ranges of 0.01 to 1.00μg/mL with the corresponding correlation coefficient≥0.9999.The limit of detection(LOD)and limit of quantification(LOQ)of the method were 2 and 10μg/kg,respectively.The average recoveries of zearalenone at supplemental levels of 0.01,0.02,0.10 and 1.00 mg/kg were 86.90% to 100.27%,with the intra-day relative standard deviations(RSD)were 3.18% to 9.88%,and inter-day RSD were 4.94% to 9.40%.In conclusion,compared with the traditional solid-phase extraction and immunoaffinity column clean-up methods,immunomagnetic beads technique integrates the extraction,separation and purification in a single run,it can achieve fully automated processing,which improve the work efficiency and avoiding result deviations caused by manual operation,it is much more suitable for the determination of zearalenone content in a large-scale number of feed samples.

关 键 词：免疫磁珠净化 玉米赤霉烯酮 真菌毒素 饲料 高效液相色谱 

分 类 号：S816.17[农业科学—饲料科学]