Determination of Biogenic Amines at Low μg L-1 Levels as Acetylacetone Derivatives by RP-HPLC with UV-Visible Detection in Expired Apple Juice Samples

Abstract

An accurate and sensitive method for trace quantification of three biogenic amines in expired apple juice samples based on reversed-phase liquid chromatography (RP-HPLC) with UV-visible detection is described. Biogenic amines including cadaverine, histamine, and tyramine, were converted to their acetylacetone derivatives in water-methanol medium. The proposed method involves a pre-column derivatization of species followed by RP-HPLC separation with Thermo Hypersil Gold reversed-phase column and UV detection at 315 nm. A flow rate of 0.9 mL min^-1 was used in the proposed method. An efficient separation of biogenic amines was successfully performed in 11 min with a good resolution using 35:65 % (v/v) acetonitrile-water mixture as the mobile phase. Detection limits of 0.03, 0.23, and 0.08 μg L^-1 were obtained for cadaverine, histamine and tyramine, respectively. The proposed method has been successfully applied for the analysis of two different commercially available expired apple juice samples. Recovery rates between 98.78 and 102.12 % were obtained with an RSD of 0.16-1.65 % for the analysis of 20 mL of expired apple juice samples indicating that the recoveries of biogenic amines were very satisfactory.