高效液相色谱法测定食品中黄曲霉毒素的方法研究

DETERMINATION OF AFLATOXIN B1,B2,G1,G2,M1 IN FOODS BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY WITH SOLID PHASE EXTRACTION.

目的 :建立一种快速、灵敏、简便的测定食品中黄曲霉毒素 (AFT) B1、 B2、 G1、 G2、 M1的高效液相色谱方法。方法 :用乙腈 -水 (9+1)提取样品中的 AFT,超声波萃取 ,再经 Myco Sep# 2 2 4或 # 2 2 6毒素专用多功能固相柱(MFC)分离纯化 ,纯化液经浓缩挥干后 ,加三氟乙酸 (TFA)衍生 ,用反相 HPL C分离、荧光检测器测定 AFT含量 ,样品与标准比较 ,保留时间定性 ,峰面积定量。结果 :五种毒素可在 10 min内完成分离 ,最低检出限可达 0 .0 2 ng,用本法对面条、食用油、花生酱、奶粉样品分别进行 5 μg/ kg、 10 μg/ kg两个水平的加标回收实验 ,A FT回收率为 78.6 %～ 10 9.9% ,加标样品 6次平行测定的 RSD为 0 .6 3%～ 2 .8% ,线性范围为 0 .15 ng～ 7.5 ng,r>0 .9988。结论 :建立了一种快速、灵敏、简便的以超声波萃取、经固相柱净化、用荧光检测器测定食品中黄曲霉毒素 (AFT) B1、 B2、G1、G2。

Objective:A rapid,sensitive and simple method for the determination of aflatoxin B1,B2,G1,G2,M1 in foods by high performance liquid chromatography with Multifunctional Cleaning Column(MFC) is reported.Method:Sample was extracted with 90% acetonitrile and cleaned-up with the MFC.The purified solution was concentrated and evaporated to dryness.Aflatoxins were derivatized with TFA and isolated by Reverse Phase HPLC.Aflatoxins were detected and quantitated by HPLC with fluorescence detection.Results:The five toxins could be separated in 10 minutes and the detection limit was 0.02ng for each toxin.The method has been successfully applied to determination of the noodles,peanut oil,peanut butter and milk powder samples.The linear range of the method is 0.15ng^7.5ng for aflatoxin B1,B2,G1,G2,and 0.18ng^9.15ng for aflatoxin M1,the correlation coefficient≥0.9988.Conclusion:The recoveries of aflatoxins spiked at 5～10×10 -9 levels was in the range of 78.6%～109.9% and the relative standard deviation was 0.63%～2.8%.