Determination of aflatoxin m1 in milk by the two methods of immunochromatography and hplc

Elahe sadat Alavi razavi ravari,1 Bahram daraei,2,* Mohammad javad rasaee,3

1. Department of Toxicology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
2. Department of Toxicology and Pharmacology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran.
3. Department of Biohemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.



Aflatoxin m1 (afm1) is a monohydroxylated derivative of aflatoxin b1 (afb1) which is secreted into milk after intake of the food contaminated with afb1. the major effects of afm1 include carcinogenesis, especially on liver; mutagenesis; and adduct formation with dna. the main objective of this research was to introduce and validate a rapid, inexpensive, and simple method based on immunochromatography as opposed to the time consuming and costly method of high performance liquid chromatography (hplc).


In order to determine afm1 presence in forty milk samples, the results that obtained by rapid test are confirmed with the reference method. the basis of this rapid test is an immunochromatography mechanism in which afm1 molecules in the sample competes with the fixed afm1-bsa on the membrane (t zone) to combine the red complex of specific immunogold afm1 monoclonal antibodies and finally, prompts the formation (negative) or lack of formation (positive) of visible region in the test area.


based on achieved results, afm1 rapid test validation had sensitivity and specificity of 100%. the results of afm1 presence determination, indicated significant correlation (p<0.01) between rapid test and reference method. among all evaluated samples, only three samples of manufactured milk had concentration more than permissible limit (100 ng/l).


Given the achieved results, we can use this rapid test designed by rojan azma co, that it’s cut-off based on the maximum permissible limit of 100 ng/l set by the institute of standards and industrial research of iran, as a reliable method for screening manufactured milk.


Immunochromatography, hplc, aflatoxinm1, milk