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Abstract
Separation and determination of mutagenic and carcinogenic heterocyclic amines by a reversed phase isocratic HPLC method is here in described. Five of them, 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-3,4,7,8-tetramethylimidazo[4,5-f] quinoxaline (4,7,8-TriMeIQx) and 2-amino-l-methyl-6-phenylimidazo[4,5-b] pyridine (PhIP) were encountered in this study. Separation was accomplished on a reverse phase HPLC column of Supelcosil LC-8 equipped with a Supelguard LC-8 precolumn, at a flow rate of 2.0 mL/min. IQ, MelQx and 4,8-DiMeIQx were resolved using 15% (v/v) acetonitrile in 50 mM triethylamine-phosphate buffer, pH 3.2, whereas 4,7,8-TriMeIQx and PhIP using 30% (v/v) acetonitrile in the same buffer. Detection of the eluted heterocyclic amines was performed at 263 nm for IQ, MeIQx and 4,8-DiMeIQx and at 254 nm for 4,7,8-TriMeIQx and PhIP.