单克隆抗体亲和层析柱在测定酱油与乳腐中黄曲霉霉素B_1的应用

APPLICATION OF MONOCLONAL ANTIBODY AFFINITY COLUMN TO MEASUREMENT OF AFLATOXIN B_1 IN SOY SAUCE AND FERMENTED BEAN CURD

本文介绍了单克隆抗体系和层析在简单、快速及有效地分离酱油及乳腐中黄曲霉霉素B_1(AFB_1)的方法。两食品经氯仿粗提后,挥干氯仿,用5％甲醇水(体积比)溶解残余物,加至单抗亲和层析柱,PBS溶液洗除杂质,最后用80％甲醇水洗脱并挥干。用TLC方法测定AFB_1含量。按10μg/L量加入酱油测定回收率,所得回收率为81.1％。本方法最低检出极限为0.2μg/L。在56份酱油样品,45份乳腐样品的AFB_1测定中,我们发现酱油中AFB_1污染水平最高,其中最高者为188μg/L,16％的酱油样品高于20μg/L。

A simple,rapid and efficient method using monoclonal antibody affinity columnis described for isolation and purification of AFB. in soy sauce and fermented bean curd.Samples were extracted with chloroform. After filtration,the chloroform was evaporated.The residue was dissolved in 5% methanol-water (v/v) and placed on a monoclonal antibody affinity column. The column was washed with PBS buffer to remove nonspeci-tical bound materials. The column was eluted with 80% methanol-water (v/v) and theelute was evaporated. AFB_1 was determined by TLC. The recovery of AFB_1 (10μg/L)was81. 1% and the limit of determination was 0. 2μg/L.Applying this technique,wefound highincidence and levels of AFB-1 contamination in soy sauce. 16% of tested samplescontained>20μg/L, and the maximum was 188μg/L,the level in fermented bean curdwas lower.