High-performance liquid chromatographic determination of biogenic amines in wines with an experimental design optimization procedure Original Research Article

A novel and sensitive HPLC method for determining biogenic amines in wine samples is described. It involves pre-column labeling of the analytes with 1,2-naphthoquinone-4-sulfonate (NQS) and liquid–liquid extraction of derivatives with chloroform for analyte preconcentration and sample clean-up. A linear gradient elution consisting of a mixture of 2% of acetic acid aqueous solution and methanol is used to separate the amine derivatives in a C18 column. The eluted compounds are detected spectrophotometrically at 270 nm. The optimization of both derivatization and separation conditions is accomplished by means of factorial and central composite designs and multicriteria decision functions. The analytical parameters of the method are established using red wine samples. Detection limits range from 0.006 to 0.315 mg L−1. The run-to-run repeatabilities of retention times and peak areas are around 0.6 and 5.6%, respectively. Recoveries ranging from 91.9 to 105% prove the accuracy of the method for determining histamine, putrescine, cadaverine, tryptamine, phenylethylamine and serotonin in red wines. The proposed method has been applied to the analysis of commercial wines from different Spanish regions.