高效液相色谱法同时检测粮食中常见8种真菌毒素的含量

Simultaneous Analysis of 8 Mycotoxins in Grains by High Performance Liquid Chromatography

建立免疫亲和柱净化-高效液相色谱法同时测定粮食中黄曲霉毒素B1(aflatoxins,AFB1)、黄曲霉毒素B2(AFB2)、黄曲霉毒素G1(AFG1)、黄曲霉毒素G2(AFG2)、赭曲霉毒素A(ochratoxin A,OTA)、玉米赤霉烯酮(zearalenone,ZEN)、呕吐毒素(deoxynivalenol,DON)和T-2毒素的检测方法。样品经乙腈-水提取后,用免疫亲和柱净化,Agilent Elipse Plus C18(100 mm×4 mm,3.5μm)色谱柱分离,以甲醇-乙腈-水-乙酸为流动相,流速1 m L/min,柱温35℃,进样量20μL,检测系统为可变波长检测器串联光化学衍生器串联荧光检测器。根据信噪比为3的峰响应值,确定各真菌毒素的检出限为:AFB1 0.446 ng/m L、AFB2 0.152 ng/m L、AFG1 0.523 ng/m L、AFG2 0.334 ng/m L、ZEN 7 ng/m L、OTA 0.7 ng/m L、DON 200 ng/m L、T-2毒素100 ng/m L。样品中各真菌毒素的平均加标回收率,玉米为80.0%~104.5%,小麦为83.2%~102.8%,方法精密度为2.6%~10.2%。从样品前处理到分析整个过程耗时约2 h。本方法简便、快速、灵敏度高,适用于粮食中多种真菌毒素的快速测定。

A rapid analytic method using high performance liquid chromatography(HPLC) was established for the simultaneous determination of aflatoxins, including AFB1, AFB2, AFG1 and AFG2, ochratoxin A, zearalenone, deoxynivalenol and T-2 toxin in grains. The samples were extracted with acetonitrile-water, and cleared up with an immunoaffinity column(IAC). The separation of the targeted compounds was performed on an Agilent Elipse Plus C18 column(100 mm×4 mm, 3.5 μm) using a mobile phase consisting of a mixture of acetonitrile : methanol : water : acetic acid with a flow rate of 1 m L/min at 35 ℃. The injection volume was 20 μL. The detection system was variable wavelength detector(VWD)-PHRED system-fluorescence detector(FLD). The limits of detection(LODs) of AFB1, AFB2, AFG1, AFG2, ZEN, OTA, DON, and T-2 toxin were 0.446, 0.152, 0.523, 0.334, 7, 0.7, 200, and 100 ng/m L, respectively. The recoveries of samples spiked with these mycotoxins were in the ranges of 80.0%- 104.5% for corn and 83.2%- 102.8% for wheat. The precision values associated with the method, expressed as relative standard deviation(RSD) values, were between 2.6% and 10.2%. It took only 2 hours to complete an analysis. The method was suitable for the determination of common 8 mycotoxins in raw grains, with the advantages of simplicity, rapidness, sensitivity, and good reproducibility.