Simultaneous determination of six non‐polar heterocyclic amines in meat samples by supercritical fluid extraction–capillary electrophoresis under fluorimetric detection

Abstract

A novel, sensitive and selective method for the separation and quantification of a group of non-polar heterocyclic amines (9H-pyrido-[3,4-b] indole, norharmane; 1-methyl-9H-pyrido-[3,4-b] indole, harmane; 2-amino-9H-pyrido-[2,3-b] indole, AalphaC; 2-amino-3-methyl-9H-pyrido-[2,3-b] indole, MeAalphaC; 3-amino-1,4-dimethyl-5H-pyrido-[4,3-b] indole, Trp-P-1 and 3-amino-1-methyl-5H-pyrido-[4,3-b] indole, Trp-P-2) in commercial meat samples has been developed. This methodology is faster than others previously described. The method is based on the combination of a supercritical fluid extraction procedure, followed by the analysis of the extracted plug by CE with fluorescence detection. The supercritical fluid extraction procedure was optimized for the clean-up of the samples and the extraction of the analytes. For the electrophoretic separation, the effect of composition, pH and concentration of buffer, organic modifier content, pressure and time of injection, capillary temperature and voltage applied were studied. A 10 mmol/L formic acid-ammonium formate-ACN (10%, v/v) solution at pH 1.5 was selected as the running electrolyte. With 5-s hydrodynamic injection, linear responses in the range from 100 to 1000 ng/mL and detection limits ranging from 15.9 to 28.1 ng/mL were obtained for different amines in less than 13 min. ACN-water (1:1 in volume) was used as a sample solvent. Fluorescence detection enhances the sensitivity and avoids interferences coming from non-fluorescent compounds present in the matrices of the sample extracts.