Determination of biogenic amines in wines by high-performance liquid chromatography with on-column fluorescence derivatization

An on-column fluorometric derivatization method was developed for determining eight biogenic amines by HPLC and fluorometric detection. The derivatization was carried out by pumping the o-phthalaldehyde reagent through the column together with other solvents used as mobile phases. The on-column reaction conditions (pH, berate and o-phthalaldehyde concentrations) were optimized. The matrix effect, the thiol used in the derivative reagent and the effect of the temperature on the separation process were also studied. Solid-phase extraction was applied prior to the derivatization procedure as a clean-up of wines by using 2 different commercially available SAX and C-18 cartridges to improve the accuracy and the precision of the method. The linearity range (0.5-15 mg l(-1)) and detection limits (between 100 and 300 mu g l(-1)) of this method were similar to the standard pre-column automated o-phthalaldehyde derivatization method. The method was checked with several red wines from Tarragona by comparing the results with the standard pre-column method.