An improved analytical method for determination of heterocyclic amines in chicken legs

Several extraction, separation and detection methods for heterocyclic amines (HAs) in chicken legs were evaluated by liquid chromatography. Results showed that the most appropriate extraction method includes the removal of macrosubstances by centrifugation and subsequent purification using a PRS (propylsulfonic acid silica gel) and a C(18) cartridge, and the recovery obtained ranged between 51 and 89%. For HPLC separation, a binary solvent system consisting of acetonitrile and 0.05 M ammonium acetate solution (pH 3.6) with gradient elution with flow rate of 1.0 mt min(-1) and detection at 258 nm was used to resolve 16 HAs. With fluorescence nine HAs could be detected by employing a programmable wavelength, and the sensitivity was 108-400 times higher than that by UV detection. The detection limits for UV and fluorescence detection were 0.02 similar to 0.5 ng and 0.05 similar to 3 pg respectively, with a signal-to-noise ratio 3. The presence of HAs in fried chicken legs was also determined.