Isolation, detection, and quantification of aflatoxin-albumin adducts in serum of rats treated with aflatoxin B1

Aflatoxins, especially aflatoxin B1, have lethal effects on human and animal health. This study is intended to present a specific, sensitive, and relatively fast method for measurement, detection, and isolation of aflatoxin-albumin (Af-Alb) adducts in serum. In this experimental-trial, three groups of rats were selected and used as positive control (treated with aflatoxin B1), negative control (without treatment) and standard (treated with radioactive aflatoxin B1). After drawing blood samples from the rats, blood serum and then, serum albumin were isolated. Albumin was hydrolyzed by pronase and eventually, was injected into HPLC system. The sample was then identified and measured by fluorescence detector. Electrophoresis on PAGE revealed albumin isolated from serum to be perfectly pure. In HPLC method, detection limit for the measurement of Af-Alb adduct was determined to be 60 pg/ml. The mean of aflatoxin positive control rats serum was 19.2 ng/mg albumin. In inter- and intra-group experiments, a remarkable level of reproducibility was seen for this method. The amount of Af-Alb adduct is proportionate to the amount of aflatoxin received. This project was conducted with rat serum sample, but since albumin is hydrolyzed and can be isolated from aflatoxin, this method is applicable to the measurement of Af-Alb adducts in human serum samples.