Tabari Mahsa, Karim Guity, Ghavami Mehrdad, Chamani Mohammad
Department of Food Science and Technology, Faculty of Agriculture and Natural Resources, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Toxicol Ind Health. 2011 Aug;27(7):629-35. doi: 10.1177/0748233710394236. Epub 2011 Mar 8.
Yoghurt is a popular dairy product in Iran because of its beneficial influence on human health and nutritional value. Aflatoxin M1 (AFM1) is the metabolite of potential carcinogen aflatoxin B1, which can contaminate milk through the feed and is not eliminated by common processing heat treatment. An analytical method using immunoaffinity column for extraction and a high-performance liquid chromatography (HPLC) for quantification was developed for AFM1 in this study. An HPLC method with fluorimetric detection for the determination of AFM1 in yoghurt milk has been optimized and validated according to Commission Decision BS EN ISO 14501: 2007 by using the conventional validation approach. The procedure for determining selectivity, recovery, precision, decision limit (CCα) and detection capability (CCβ) of the method has been reported. The results of the validation process demonstrate the agreement of the method with the provisions of Commission Regulation 401: 2006:EC. A new HPLC method with fluorescence detection was developed to determine aflatoxin M1. The detection limit was 1 ng/kg for yoghurt. The calibration curve was linear from 0.1 to 3.0 μg l⁻¹ injected. The method includes a preliminary clean-up and the average recoveries determined on three different days at the concentration levels of 0.025, 0.050 and 0.075 μg kg⁻¹ were in the range of 72.57%-86.66% with RSD in the range of 2.56%-8.41%. The interday and interlevel mean recovery value, which has been used to correct routine analysis results, was 80%. The method is rapid, easily automatable and therefore useful for accurate and precise screening of aflatoxin M1 in yoghurt.
酸奶因其对人体健康的有益影响和营养价值，在伊朗是一种受欢迎的乳制品。黄曲霉毒素M1（AFM1）是潜在致癌物黄曲霉毒素B1的代谢产物，它可通过饲料污染牛奶，且普通加工热处理无法将其去除。本研究开发了一种使用免疫亲和柱萃取和高效液相色谱法（HPLC）定量检测AFM1的分析方法。已根据委员会决定BS EN ISO 14501:2007，采用传统验证方法对用于测定酸奶中AFM1的带荧光检测的HPLC方法进行了优化和验证。报告了该方法的选择性、回收率、精密度、决策限（CCα）和检测能力（CCβ）的测定程序。验证过程的结果表明该方法符合委员会法规401:2006:EC的规定。开发了一种新的带荧光检测的HPLC方法来测定黄曲霉毒素M1。酸奶的检测限为1 ng/kg。进样浓度在0.1至3.0 μg l⁻¹范围内时校准曲线呈线性。该方法包括初步净化，在0.025、0.050和0.075 μg kg⁻¹浓度水平下，在三个不同日期测定的平均回收率在72.57% - 86.66%范围内，相对标准偏差在2.56% - 8.41%范围内。用于校正常规分析结果的日间和不同水平平均回收率值为80%。该方法快速、易于自动化，因此可用于准确、精确地筛查酸奶中的黄曲霉毒素M1。
