Evaluation of Human Exposure to Food-borne Heterocyclic Aromatic Amines using UPLC-MS/MS
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Abstract:
An accurate and highly sensitive assay was developed for the determination of sulfinamide adducts formed by heterocyclic aromatic amines (HAAs) and human serum albumin (HSA) after human ingestion and absorption, using ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The aim of this study was to assess exposure to HAAs, and further investigate the related cancer risks. Using 2-amino-1-methyl-6-phenylimidazo [4,5-b] pyridine (PhIP) as a research subject, the PhIP-HSA sulfinamide adduct was synthesized in vitro according to the metabolic pathway of the human body and was quantified using UPLC-MS/MS based on the detection of PhIP derived from its acid hydrolysis. The results showed that the recovery of HSA by the HiTrap Blue affinity columns was greater than 90%, and the hydrolysis efficiency of the PhIP-HSA sulfinamide adduct reached 96% under optimized conditions. There was a positive linear relationship between the PhIP-HSA sulfinamide adduct content and the signal intensity of PhIP, and the linear regression equation was y=1.011 7x+3.256 3, R²=0.998 7. The respective detection and quantitation limits for the PhIP-HSA sulfinamide adduct in plasma were as low as 5×10-3 fmol/mg HSA and 1.50×10-3 fmol/mg HSA. This method showed a high degree of sensitivity when determining PhIP-HSA sulfinamide adducts in human plasma, and proved to be a feasible method for the evaluation of exposure to HAAs in individuals who frequently eat cooked meat, as well as the associated cancer risks.
