Determination of Biogenic Amines in Wine Using Modified Liquid-Liquid Extraction with High Performance Liquid Chromatography-Fluorescence Detector

Abstract

This study aimed to establish a reliable method to determine the level of eight biogenic amines in wine using derivatization and modified liquid-liquid extraction with high performance liquid chromatography-fluorescence detector (HPLC-FLD). HPLC separation conditions were optimized to analyze the derivatized biogenic amines. The standard curve of each biogenic amine exhibited a good linearity confirmed by determination coefficient (0.9983–0.9999) under the concentration range of 0.1 to 50.0 mg/L. The intra- and inter-day precision of the mixed standard (15 mg/L) appeared to be 4.86 and 5.22%, respectively. The limit of detection and limit of quantification ranged of 0.001–0.050 mg/L and 0.005–0.167 mg/L, respectively. To achieve the optimal extraction, 1.0 mL of wine sample was derivatized with 1000 μL of dansyl chloride followed by derivatization termination using 100 μL of ammonium hydroxide. The derivatized amines were further extracted with 1.7 mL of acetonitrile and then centrifuged at 10,000 rpm for 5 min. The optimized extraction was further validated in two wine samples spiked with these biogenic amines under three concentration levels (1.0, 5.0, and 10.0 mg/L). The recoveries of these target amines were found to be 73.02 to 111.61%. A total of 57 wine samples were analyzed using this established method to determine their biogenic amines level. It was found that the red wine samples contained higher concentration than the white wines. The predominant biogenic amines found in these wines included putrescine, tyramine, and 2-phenylethylamine. This study could provide a reliable and accurate method on the determination of the concentration of biogenic amines in wine.