The invention relates to an innovative modified immunoassay of the competitive type, using polyclonal, monoclonal or recombinant antibodies, for the quantitative determination of Aflatoxin M1 in milk or liquid samples of milk composition. The invention provides procedures for the highest recovery quantitation of Aflatoxin M1 which utilize graded addition of Aflatoxin M1 antibody configuration and orienting factors during the binding process of the antibody to the solid support and standard solutions of powdered skim instant-dissolving milk containing non-zero fat acids. The immunochemical construction includes the indirect process of connecting the antibodies to the solid substrate, through a covalent or strong non-covalent bond with molecules of protein nature directly immobilized on the solid substrate and the graded addition of nonionic surfactants, polymers of low molecular weight and low degree of hydrolysis and disaccharides. This innovative modified immunoassay system quantifies Aflatoxin M1 with high recovery, in a range of 0-250 ng/L.
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