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使用免疫亲和色谱法作为测定奶酪中黄曲霉毒素M1的纯化步骤。

Use of immunoaffinity chromatography as a purification step for the determination of aflatoxin M1 in cheeses.

作者信息

Dragacci S, Gleizes E, Fremy J M, Candlish A A

机构信息

Ministere de l'Agriculture et de la Pêche, Centre National d'Etudes Veterinaires et Alimentaires, Laboratoire Central d'Hygiene Alimentaire CNEVA-LCHA, Paris, France.

出版信息

Food Addit Contam. 1995 Jan-Feb;12(1):59-65. doi: 10.1080/02652039509374279.

Abstract

A method using immunoaffinity as a purification step for the determination of aflatoxin M1 in cheese is described. A simple solvent extraction with dichloromethane followed by a washing step with N-hexane gives a prepurified extract. A comparison between two ways of aflatoxin M1 purification, by solid-phase extraction clean-up and by immunoaffinity, was carried out. The use of immunoaffinity columns containing monoclonal antibodies against aflatoxin M1 gives the best result, i.e. a very clean preparation containing purified aflatoxin M1. The quantification of aflatoxin M1 is then performed by high performance liquid chromatography using fluorometric detection. This method was successfully carried out on naturally-contaminated and spiked cheeses. Recoveries are about 75%. The limit of quantification is 0.020 microgram of aflatoxin M1 per kg of cheese. This method seems suitable for use in monitoring programmes for aflatoxin M1 contamination in dairy products such as cheese.

摘要

描述了一种使用免疫亲和作为纯化步骤来测定奶酪中黄曲霉毒素M1的方法。先用二氯甲烷进行简单的溶剂萃取,然后用正己烷洗涤,得到预纯化提取物。对黄曲霉毒素M1的两种纯化方法进行了比较,即固相萃取净化和免疫亲和。使用含有抗黄曲霉毒素M1单克隆抗体的免疫亲和柱可得到最佳结果,即得到含有纯化黄曲霉毒素M1的非常纯净的制剂。然后通过高效液相色谱法和荧光检测对黄曲霉毒素M1进行定量。该方法已成功应用于天然污染和添加了黄曲霉毒素M1的奶酪。回收率约为75%。定量限为每千克奶酪0.020微克黄曲霉毒素M1。该方法似乎适用于奶酪等乳制品中黄曲霉毒素M1污染的监测计划。

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