Extraction, detection, and quantification of heterocyclic aromatic amines in Portuguese meat dishes by HPLC/diode array

Abstract

The present paper describes the results obtained in the analyses of heterocyclic aromatic amines (HAs) in bovine meat dishes prepared by three different coking methods; usually ingredients such as, salt, garlic, wine, olive oil, onion, and tomato were added. Control meat samples (cooked without ingredients) were also prepared. Analyses were performed by HPLC/diode array and detection of the eluted HAs was performed at 263nm. The calibration graphs for fourteen amines injected into the column were linear up to approximately 2.0ng/mu L and the detection limits (signal-to-noise ratio 3:1) ranged from 0.06 to 0.4ng injected. Nine HAs, namely 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4,8-DiMeIQx), 2-amino-1-methyl-6-phenylimidazol[4,5]pyridine (PhIP), 3-amino-1,4-dimethyl-5H-Pyrido[4,3-b]indole (Trp-P-1), 3-amino-1-methyl-5H-Pyrido[4,3-b]indole (Trp-P-2), 2-amino-9H-Pyrido [2,3-b]indole (AC) 2-amino-3-methyl-9H-Pyrido [2,3-b]indole (MeAC), 2-amino-6-methyldipyrido[1,2-a:3',2'-d]imidazole (Glu-P-1) were encountered in meat samples. A paired Student's t-test showed no significant differences on HAs levels (p > 0.05) of control meat samples (cooked without ingredients) and meat samples cooked with ingredients usually in the Portuguese diet and rich in antioxidants. Changes in cooking habits are needed to make possible the use of ingredients not only as flavoring but also as protective agents against HAs formation during domestic cooking.