摘要
目的开发快速检测方法检测木薯叶粉中的赭曲霉毒素A(Ochratoxins A,OTA)。方法利用单克隆抗体和酶标抗原,建立了竞争酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)快速检测方法。用不同浓度的氯化钠甲醇水溶液、乙腈和磷酸盐缓冲液等作为样本提取液提取木薯叶粉中OTA,用活性炭和稀释法减少样本基质干扰。结果ELISA灵敏度为0.2 ng/mL,IC50为0.878 ng/mL。木薯叶粉用含氯化钠的70%甲醇水溶液提取,离心后取上清5倍稀释,样本回收率80%~121%,检测限为5μg/kg,满足国标规定食品中OTA不超过5μg/kg的要求,检测时间为30 min,一次可批量检测90个样本。结论用本研究建立的ELISA方法检测木薯叶粉及其食品中的OTA含量结果准确、操作简单、成本低、时间短、重复性好,适合基层批量筛查。
Objective To develop a rapid test method for Ochratoxin A in cassava leaf powder. Methods Rapid detection of competitive enzyme-linked immunosorbent assay by using monoclonal antibody and enzyme-labeled antigen. OTA in tapioca leaf powder was extracted with different concentrations of sodium chloride methanol solution,acetonitrile and phosphate buffer,etc. Activated carbon and dilution method were used to reduce sample matrix interference. Results ELISA sensitivity was 0.2 ng/mL and IC50 was 0.878 ng/mL. Cassava leaf powder was extracted with 70% methanol solution containing sodium chloride. After centrifugation,the supernatant was diluted 5 times. The sample recovery was 80%-121%. The detection limit was 5 μg/kg,which meet the requirements of the national standard that OTA in food should not exceed 5 μg/kg. The detection time was 30 min,and 90 samples can be detected in batches at a time. Conclusion The ELISA method established in this study can be used to detect OTA content in tapioca leaf powder and its food with accurate results,simple operation,low cost,short time,good repeatability,and suitable for mass screening at the grass-roots level.
作者
余厚美
王琴飞
林立铭
徐缓
张振文
YU Hou-Mei;WANG Qin-Fei;LIN Li-Ming;XU Huan;ZHANG Zhen-Wen(Key Laboratory of Ministry of Agriculture for Germplasm Resources Conservation and Utilization of Cassava,National R&D Centre for Potato Processing,Tropical Crops Genetic Resources Institute,Chinese Academy of Tropical Agricultural Sciences,Danzhou 571737,China)
出处
《食品安全质量检测学报》
CAS
2019年第24期8242-8250,共9页
Journal of Food Safety and Quality
基金
中国热带农业科学院基本科研业务费专项资金(1630032019017)
现代农业产业技术体系(CARS-12)专项资金
中国热带农业科学院热带作物品种资源研究所科技成果转移转化专项资金(PZS2019009)~~
关键词
木薯叶粉
木薯
赭曲霉毒素A
快速检测
Cassava leaf powder
Cassava
Ochratoxins A
rapid detection
