Chu F S, Lee R C, Trucksess M W, Park D L
Food Research Institute, University of Wisconsin-Madison 53706.
J Assoc Off Anal Chem. 1988 Sep-Oct;71(5):953-6.
A simple, rapid enzyme-linked immunoassay (ELISA) was used to evaluate the performance of each step (extraction, filtration, solvent partition, and silica gel column chromatography) of a solvent-efficient thin-layer chromatographic (TLC) method which is undergoing interlaboratory collaborative study for the determination of aflatoxin B1 in corn, raw peanuts, and peanut butter. The apparent average recoveries using the ELISA method were about 30 to 50% higher than those using the TLC method if only the amount of B1 added to the samples was used in the calculations. After the cross-reaction of the antibody with other aflatoxins added to the samples was considered, the amounts recovered approached the levels of aflatoxins added in all 3 commodities tested. With no cleanup treatment, ELISA recoveries at aflatoxin B1 levels above 7.5 ng/g were 84, 79, and 103% for corn, raw peanuts, and peanut butter, respectively. The coefficients of variation were between 5.2 and 25.2%. With each cleanup step in the TLC method, ELISA detected a progressive decrease in recovery from 150.5 to 105.3% (before correction for the presence of other aflatoxins) or from 93.5 to 65.4% (after correction for other aflatoxins) of B1 added to the samples. The ELISA data support the conclusion obtained from previous studies that cleanup treatments were not necessary in the ELISA. When large amounts of other aflatoxins are present, an understanding of the cross-reactivity of antibody with other aflatoxins in the ELISA is essential for final interpretation of the data.
一种简单、快速的酶联免疫吸附测定法（ELISA）被用于评估一种溶剂高效薄层色谱法（TLC）每个步骤（提取、过滤、溶剂分配和硅胶柱色谱）的性能，该TLC方法正在进行实验室间协作研究，用于测定玉米、生花生和花生酱中的黄曲霉毒素B1。如果仅在计算中使用添加到样品中的B1量，使用ELISA方法的表观平均回收率比使用TLC方法的回收率高约30%至50%。在考虑抗体与添加到样品中的其他黄曲霉毒素的交叉反应后，回收量接近所有3种测试商品中添加的黄曲霉毒素水平。在没有净化处理的情况下，黄曲霉毒素B1水平高于7.5 ng/g时，玉米、生花生和花生酱的ELISA回收率分别为84%、79%和103%。变异系数在5.2%至25.2%之间。随着TLC方法中每个净化步骤的进行，ELISA检测到添加到样品中的B1回收率逐渐下降，从未校正其他黄曲霉毒素存在时的150.5%降至105.3%，或在校正其他黄曲霉毒素后从93.5%降至65.4%。ELISA数据支持先前研究得出的结论，即ELISA中不需要净化处理。当存在大量其他黄曲霉毒素时，了解ELISA中抗体与其他黄曲霉毒素的交叉反应对于最终解释数据至关重要。
