Zhang Zhaowei, Hu Xiaofeng, Zhang Qi, Li Peiwu
Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, PR China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, PR China.
Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, PR China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2016 May 15;1021:145-152. doi: 10.1016/j.jchromb.2016.02.035. Epub 2016 Feb 27.
Mycotoxins usually found in agricultural products such as peanut, corn, and wheat, are a serious threat to human health and their detection requires multiplexed and sensitive analysis methods. Herein, a simultaneous determination for aflatoxin B1, B2, G1, G2, ochratoxin A, zearalanone and T-2 toxin was investigated using high performance liquid chromatography coupled with tandem mass spectrometry in a single run via a home-made multiple immunoaffinity column. Four monoclonal antibodies were produced in our lab against aflatoxins, ochratoxin A, zearalanone and T-2 toxin, respectively, then combined as a pool and bound to Sepharose-4B for affinity chromatography. Seven mycotoxins were effectively extracted from the agricultural product samples by using acetonitrile/water/acetic acid (80:19:1, v/v/v) Then, the extraction was cleanup by multiple immunoaffinity column. This method demonstrated a considerable linear range of 0.30-25, 0.12-20, 0.30-20, 0.12-20, 0.60-30, 0.30-25, and 1.2-40μgkg(-1)and lower limits of detection at 0.1, 0.04, 0.1, 0.04, 0.2, 0.1 and 0.4μgkg(-1) for AFB1, AFB2, AFG1, AFG2, OTA, ZEN and T-2, respectively, in comparison with previously reported methods, as well as excellent recoveries. The mIAC capacity for AFB1, AFB2, AFG1, AFG2, OTA, ZEN, and T-2 were 187, 181, 153, 151, 105, 130, 88ng, respectively. It was found that all of the 7 mycotoxins were present in 90 agricultural product samples. The proposed method meets the requirements for rapid sample preparation and highly sensitive identification of multiple mycotoxins in agricultural product and food safety. This method provides a promising alternative with high throughput and high sensitivity for rapid analysis of seven mycotoxins in the monitoring of food safety.
通常存在于花生、玉米和小麦等农产品中的霉菌毒素对人类健康构成严重威胁，其检测需要多种且灵敏的分析方法。在此，研究了使用高效液相色谱-串联质谱联用技术，通过自制的多重免疫亲和柱在一次运行中同时测定黄曲霉毒素B1、B2、G1、G2、赭曲霉毒素A、玉米赤霉烯酮和T-2毒素。在我们实验室分别制备了针对黄曲霉毒素、赭曲霉毒素A、玉米赤霉烯酮和T-2毒素的四种单克隆抗体，然后将它们混合并与琼脂糖-4B结合用于亲和色谱。使用乙腈/水/乙酸（80:19:1，v/v/v）从农产品样品中有效提取七种霉菌毒素。然后，通过多重免疫亲和柱对提取物进行净化。该方法显示出相当宽的线性范围，分别为0.30 - 25、0.12 - 20、0.30 - 20、0.12 - 20、0.60 - 30、0.30 - 25和1.2 - 40μgkg(-1)，黄曲霉毒素B1、B2、G1、G2、赭曲霉毒素A、玉米赤霉烯酮和T-2毒素的检测下限分别为0.1、0.04、0.1、0.04、0.2、0.1和0.4μgkg(-1)，与先前报道的方法相比，回收率也很高。多重免疫亲和柱对黄曲霉毒素B1、B2、G1、G2、赭曲霉毒素A、玉米赤霉烯酮和T-2毒素的容量分别为187、181、153、151、105、130、88ng。发现90个农产品样品中均存在所有7种霉菌毒素。所提出的方法满足农产品中多种霉菌毒素快速样品制备和高灵敏度鉴定以及食品安全的要求。该方法为食品安全监测中七种霉菌毒素的快速分析提供了一种具有高通量和高灵敏度的有前景的替代方法。
