反相液相色谱法测定及确证奶酪中的黄曲霉毒素M1

Reverse phase liquid chromatographic determination and confirmation of aflatoxin M1 in cheese.

作者信息

Hisada K, Terada H, Yamamoto K, Tsubouchi H, Sakabe Y

出版信息

J Assoc Off Anal Chem. 1984 May-Jun;67(3):601-6.

Abstract

A systematic method is proposed for determination and confirmation of aflatoxin M1 in cheese by liquid chromatography (LC). A sample of cheese is extracted with chloroform, cleaned up on 2 silica gel columns followed by a Sep-Pak C18 cartridge, and chromatographed on a 5 microns octadecyl silica column with fluorometric detection. The sample extract or standard is treated with n-hexane-trifluoroacetic acid (TFA) (4 + 1) for 30 min at 40 degrees C. Analysis by LC with TFA-treatment of the extract provides quantitative data. Multiple assays of 5 samples of Gouda cheese spiked with aflatoxin M1 at levels of 0.5, 0.1, and 0.05 ng/g showed average recoveries of 93.2, 91.6, and 92.4%, with coefficients of variation of 2.63, 3.97, and 4.52%, respectively. Assay of 5 naturally contaminated cheeses resulted in 0.051-0.448 ng/g of aflatoxin M1. Limit of quantitation is about 0.01 ng/g. The identity of aflatoxin M1 is confirmed by treating aflatoxin M1 or the M2a derivative with TFA-methanol (or ethanol) (3 + 1). The TFA-methanol reaction products of M2a could be detected quantitatively.

摘要

本文提出了一种通过液相色谱法（LC）测定和确证奶酪中黄曲霉毒素M1的系统方法。取奶酪样品用氯仿萃取，先后通过两根硅胶柱和一个Sep-Pak C18柱进行净化处理，然后在一根5微米十八烷基硅胶柱上进行色谱分析，并采用荧光检测。将样品提取物或标准品用正己烷 - 三氟乙酸（TFA）（4 + 1）在40℃处理30分钟。对提取物进行TFA处理后通过LC分析可提供定量数据。对添加了水平为0.5、0.1和0.05 ng/g黄曲霉毒素M1的5份高达干酪样品进行多次测定，平均回收率分别为93.2%、91.6%和92.4%，变异系数分别为2.63%、3.97%和4.52%。对5份天然污染奶酪的检测结果显示黄曲霉毒素M1含量为0.051 - 0.448 ng/g。定量限约为0.01 ng/g。通过用TFA - 甲醇（或乙醇）（3 + 1）处理黄曲霉毒素M1或M2a衍生物来确证黄曲霉毒素M1的身份。M2a的TFA - 甲醇反应产物可进行定量检测。