Determination of heterocyclic amines in flame-grilled fish patty by capillary electrophoresis

A simple and reliable capillary zone electrophoretic (CZE) method for quantifying mutagenic and carcinogenic heterocyclic amines (HCAs) in cooked foods with UV-visible diode-array detection is described. The buffer system consisted of 50 mM disodium hydrogen phosphate, 30 mM sodium chloride, 20 mM citric acid and 26% methanol at pH 2.1. An uncoated silica tubing, 51 cm in length, was used for the CZE separation. The capillary tubing temperature was maintained at 25 degrees C with a constant voltage of 20 kV. The reproducibility of the method was over 95% for a five-replicate analysis of 10 micrograms/l-spiked 2-amino-3,4,7,8-tetramethylimidazo[4,5-f]quinoxaline (4,7,8-TriMeIQx) and the detection limit was in the low microgram/l range with coefficients of variation between 6 and 18%. An analytical run took only 15 min for 12 known HCAs. Using this procedure, up to 30 samples could be analysed in a single day. The method is reliable and can be used for screening of various HCAs. It has been applied to assess the concentrations of heterocyclic amines in otak-otak, a Malay-style grilled fish patty. The major mutagenic contaminant found in this foodstuff was 2-aminodipyrido[1,2-a:3',2'-d]imidazole (Glu-P-2) (286-1068 micrograms/kg), followed by 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) (1.6-13.0 micrograms/kg) and 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) (14.0-87.5 micrograms/kg). Two co-mutagens norharman (NH) and harman (H) were also detected in otak-otak at levels of 2.0-13.0 micrograms/kg and 12.8-21.3 micrograms/kg, respectively. The substantial amount of Glu-P-2 detected in otak-otak is probably a result of the ingredients used and the high temperature grilling process.