Determination of heterocyclic aromatic amines by capillary high-performance liquid chromatography with diode array detection in ready-to-eat cooked ham treated with electron-beam irradiation

Heterocyclic aromatic amines (HAs) are a group of mutagenic and carcinogenic substances present in significant amounts in cooked meat and fish that can potentially be formed during food processing operations. This paper proposes a capillary liquid chromatography method with diode array detection for the trace-level determination of three HAs, namely, MeIQx (2-amino-3,8-dimethylimidazo4,5-f]quinoxaline), norharman (9H-pyrido3,4-b]indole) and harman (1-methyl-9H-pyrido3,4-b]indole), in ready-to-eat (RTE) cooked ham processed by electron-beam (accelerated electrons) irradiation to eliminate pathogenic microorganisms and to extend its shelf-life. The HAs selected have frequently been detected and quantified in a wide range of food and could be potential markers to indicate the presence of these toxic compounds. The method is based on the separation in an Inertsil C(8) capillary column (150 mm x 0.3-mm internal diameter, 3 microm) by gradient elution mode using a mixture of acetonitrile and 30 mM ammonium acetate pH 4.5 buffer as the mobile phase. Detection was at 250 and 265 nm and, to improve sensitivity, large injection volumes (20 microL) and on-column focusing techniques based on the injection of HA samples in low organic solvent strength solutions were employed. A simple and short solid-phase extraction and purification procedure was also optimized for sample preparation. Nonirradiated and irradiated RTE cooked ham samples at doses between 1 and 8 kGy were analyzed. HAs were not detected in any of the samples analyzed; so both types of samples were spiked at concentration levels in the range 5-25 ng g(-1), which may be found in meat products. The quality parameters of the method developed in the food matrix were established, and detection limits around 0.3 ng g(-1) were obtained. Spiked recoveries between 70 and 79% (n = 3 for each spiked level) relative standard deviations between 1 and 5% were also obtained, showing the effectiveness of the proposed method.