HPLC–UV method validation for the identification and quantification of bioactive amines in commercial eggs

Abstract

A quantitative and confirmatory high-performance liquid chromatography with ultraviolet detection (HPLC–UV) method for the determination of bioactive amines in the albumen and yolk of commercial eggs was developed, optimized and validated by analyte extraction with trichloroacetic acid and pre-column derivatization with dansyl chloride. Phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine standards were used to evaluate the following performance parameters: limit of detection (LoD), limit of quantification (LoQ), selectivity, linearity, precision, recovery and ruggedness. The LoD of the method was defined from 0.2 to 0.3mgkg−1 for the yolk matrix and from 0.2 to 0.4mgkg−1 for the albumen matrix; the LoQ was from 0.7 to 1.0mgkg−1 for the yolk matrix and from 0.7 to 1.1mgkg−1 for the albumen matrix. The validated method exhibited excellent selectivity and separation of all amines with coefficients of determination higher than 0.99. The obtained recovery values were from 90.5% to 108.3%, and the relative standard deviation (RSD) was lower than 10% under repeatability conditions for the studied analytes. The performance parameters show the validated method to be adequate for the determination of bioactive amines in egg albumen and yolk.