Method optimization and seperation of aromatic biogenic amines and precursor amino acids by high performance liquid chromatography

Abstract

In this study, the determination of aromatic biogen amines; histamine, β-phenyl ethylamine, tyramine and tryptamine, and their precursor amino acids; histidine, phenylalanine, tyrosine and tyriptophan was performed without using any derivatization reagents. Mobile phase was optimised by establishing relationships between retention parameters and Reichardt’s ETN scale of solvent polarity. In addition, selectivity (α) and resolution (RS) values were used in order to establish a general model relating elution behaviour of substances with the composition of mobile phase. In order to find the optimum pH value, pKa values of amino acids in the mobile phase were determined by potentiometric method in 20% and 30% (v/v) MeCN media. For biogenic amines and amino acids studied, the optimised mobile phase was composed of acetonitrile-water and 6.10-3 mol l-1 1-heptanesulphonic acid sodium salt adjusted at pH 2.45 with a percentage of MeCN-water of 28% (v/v).