光化学衍生-高效液相色谱法测定粮谷类样品中黄曲霉毒素

Determination of aflatoxins in cereal food by high performance liquid chromatography with post-column derivatization

目的建立光化学衍生-高效液相色谱荧光法测定粮谷类样品中的黄曲霉毒素（AFT）含量。方法样品经Romer Labs免疫亲和柱净化,经SW-3光化学柱后衍生,经高效液相色谱分离和荧光检测器测定,分析其中黄曲霉毒素B1、B2、G1、G2的含量。同时对免疫亲和柱洗脱条件、流动相的洗脱程序进行了优化。结果在0.5~10 ng/mL（AFT B1,G1）和0.15~3.0 ng/mL（AFT B2,G2）线性范围内,所得回归方程的相关系数均大于0.999。黄曲霉毒素B1、G1方法检出限为0.15 ng/g,黄曲霉毒素B2、G2方法检出限为0.05 ng/g,加标回收率为89.5%~107%,精密度为1.4%~7.2%。采集粮谷类样品222件,其中有5件样品检出AFT,但均未超过国家限值标准。结论该方法灵敏度和准确度较高,可适用于粮谷类食品中黄曲霉毒素的检测。

Objective To establish a method for the determination of aflatoxin B1,B2,G1 and G2 in cereal food by high performance liquid chromatography（HPLC） with post-column derivatization.Methods Samples were cleaned up by immunoaffinity column Romer Labs,post-column derivatized by SW-3,separated by HPLC,detected with fluorescence detector,and then determined for the content of aflatoxin B1,B2,G1 and G2.Meanwhile,the elution conditions of immunoaffinity column and the elution program of mobile phase were optimized.Results The correlation coefficients of linear equations were all above 0.999 in the range of 0.5~10 ng/mL of aflatoxin B1 and G1,and 0.15~3.0 ng/mL of aflatoxin B2 and G2.The limits of detection were 0.15 ng/g for aflatoxin B1 and G1,and 0.05ng/g for aflatoxin B2 and G2.The recoveries of aflatoxins were 89.5%~107%with relative standard deviations of1.30%~7.15%.A total of 222 samples were analyzed and among which 5 samples were found to contain aflatoxins,but their contents were within the national standard limits.Conclusion This method is sensitive and accurate,which can be used for the determination of aflatoxin B1,B2,G1 and G2 in cereal food.