Central Composite Design to Optimizate the Derivatization Procedure for Analysis of Biogenic Amines by HPLC-UV

A simple benzoylation procedure was optimized for the biogenic amines derivatization prior to high-performance liquid chromatography determination. The significant factors affecting biogenic amine benzoylation yield were optimized by central composite design (CCD). The obtained optimal conditions resulted in a chromatographic peaks area increase with fewer interferences. Moreover, the efficiency parameters for chromatographic separation of putrescine, histamine and tyramine were determined under optimum operating conditions, which included: capacity factors or retention (k') = 2.5-4.2, selectivity factors (α) > 1 and resolution values (Rs) > 1.5, indicating that the column provided good separation of the analytes. The optimized method was applied in spiked fish samples with standard solutions of putrescine, histamine and tyramine at concentrations of 50, 80 and 50 mg L-1, respectively, to achieve recovery rates in a range from 90.89 to 96.65% for putrescine; 93.36 to 95.87% for histamine; 86.57 to 93.33% for tyramine, with the relative standard deviation (RSD) less than 4%.