Determination of carcinogenic heterocyclic amines in satay by liquid chromatography

The successful determination of mutagenic/carcinogenic heterocyclic amines (HCAs) in trace amounts (ng/g, ppb levels) in commercial foods depends largely on the detection sensitivity and selectivity, as well as the quality of clean-up of complex food matrices. In the present study, UV and fluorescence detection in series with gradient elution HPLC was used for the separation and determination of eight HCAs present in a local delicacy, satay. The identification of peaks was carried out by UV diode-array spectrophotometry. The most common HCA found in all types of satay (chicken, mutton, pork) cooked under Chinese and Malay styles was 2-amino-9H-pyrido[2,3-b]indole (AαC) (1.3-12 ppb) while 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) was found at comparatively high levels (78 ppb) in both Chinese-style pork and Malay-style chicken satay. Other major HCAs included 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) (non-detectable (ND) to 31 ppb), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) (ND-12 ppb) and 2-aminodipyrido[1,2-a:3′,2′-d]-imidazole (Glu-P-2) (ND-25 ppb). Comutagen norharman (NH) was also detected at levels up to 84 ppb, while harman (H) was only detected in one sample (Chinese-style pork satay), at a level of 25 ppb. The differences in the type and level of HCAs in satay samples may be attributed to the variation in raw foodstuffs cooked under different styles by Chinese or Malay vendors, and the ingredients added during cooking.