Simple methods for quantifying mutagenic heterocyclic aromatic amines in food products

Two solid-phase extraction methods were developed for the determinationof mutagenic heterocyclic aromatic amines in heated meat products.The copper phthalocyanine (CPC) tandem extraction was performedon coupled cartridges of diatomaceous earth and CPC-derivatizedSephasorb HP, followed by further clean-up on Sephasorb HP.Parts per billion levels of 2-amino-3,8-dimethylimidazo4,5-f]quinoxaline(MeIQx) and its homologs as well as 2-amino-3-methylimidazo4,5-f]quinoline(IQ), 2-amino-3,8-dimethyl-imidazo4,5-f]quinoline (MeIQ), 2-amino-1-methyl-6-phenyl-imidazo4,5-b]pyridine(PhIP), amino--carboline (AC), 3-amino-1,4-dimethyl-5H-pyrido4,3-b]indole(Trp-P-1), 3-amino-1-methyl-5H-pyrido4,3-b]indole (Trp-P-2),harman (H) and norharman (NH) can then be simultaneously quantifiedby HPLC with UV detection. The propylsulfonyl silica gel (PRS)tandem extraction is a one-step clean-up method on coupled cartridgesof diatomaceous earth and PRS, suitable for the determinationof MeIQx, IQ and their homologs, as well as the glutamic acidpyrolysates 2-amlno-6-methyldipyrido1,2-a:3',2']imidazole (Glu-P-1)and 2-aminodipyrido1,2-a:3',2'-d]imidazole (Glu-P-2). 4,7,8-TriMeIQxor 7,8-DiMeIQx were used as internal standards. Four grams ofsample or less are required for analysis. The recovery of theamineswas between 46 and 83% and the detection limit was in the lowp.p.b. range with coefficients of variation ranging between5 and 18%. The major mutagenic contaminant found in meat extractswas MeIQx (from <1 to 44 p.p.b.), followed by 4,8-DiMeIQx(1.3–5 p.p.b.) whereas the major contaminant in friedmeat was PhIP (23–48 p.p.b.), followed by MeIQx (5.1–8.3p.p.b.), AC (3.2–8.9 p.p.b.) and 4,8-DiMeIQx (1.3–2p.p.b.).The co-mutagens NH and H were found in fried meat atlevels of 8.7–19 p.p.b. and 3–4.8 p.p.b. respectively.