Aflatoxin M1 is the foremost metabolite of aflatoxin B1 in humans and animals, which may be present in animal products from animals fed with aflatoxin B1 contaminated feed. In this study a high performance liquid chromatography method for determination of aflatoxin M1 in eggs was described. The egg samples were diluted with warmed water and the toxin was immunoextracted followed by fluorescence detection. The average recovery of aflatoxin M1 at the three different levels 0.05, 0.1, and 0.5？μg/kg varied between 87% and 98%. The method is linear from the limit of quantification 0.05？μg/kg up to 3？μg/kg levels. This method is intended for aflatoxin M1 analyses in eggs simply with minimum toxin lose, excellent recovery, and accurate results with the limit of detection 0.01？μg/kg. 1. Introduction Mycotoxins are toxins produced by molds that cause diseases called mycotoxicosis [1]. Aflatoxins, the most common mycotoxins, are toxic metabolites produced by certain fungi that can occur in foods and feeds. Aflatoxin M1 (AFM1) is usually considered to be a detoxication byproduct of aflatoxin B1 and it is also the hydroxylated metabolite present in animal products that eat foods containing the aflatoxin B1 toxin. Aflatoxin M1 is cytotoxic and its acute toxicity is similar to that of aflatoxin B1 [2] and was classified in Group 2B as possibly carcinogenic to humans [3]. AFM1 is very slightly soluble in water, freely soluble in moderately polar organic solvents and insoluble in non-polar solvents. AFM1 is unstable to ultraviolet light in the presence of oxygen, pH (<3, >10) and oxidizing agents. On the other hand AFM1 has an intensely fluorescent in ultraviolet light [4]; its structural formula is given in Figure 1. Processing and storage cause a little effect on AFM1 content in milk and milk products [2, 5, 6]. Figure 1: Structural formula for aflatoxin M1. AFM1 has been found worldwide in a range of animal products, including milk and milk products types, eggs, meat, and meat products [7–14]. Several chromatographic methods for AFM1 determination in various commodities include ELISA and flow-injection immunoassay system [15–17], HPLC methods with solid phase and immunoaffinity separations [18, 19], and other chromatographic post column, mass, and tandem mass spectrometry detection [20–22]. This study develops an HPLC method using immunoaffinity column with rapid and reasonable high test recovery comparable with other intended methods of analyses. 2. Materials and Methods 2.1. Chemicals and Materials All chemicals and reagents were of HPLC and analytical grade.
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