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 dete rmining 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 c oncentrations) were optimized. The matrix effect, the thiol used in th e derivative reagent and the effect of the temperature on the separati on process were also studied. Solid-phase extraction was applied prior to the derivatization procedure as a clean-up of wines by using 2 dif ferent commercially available SAX and C-18 cartridges to improve the a ccuracy and the precision of the method. The linearity range (0.5-15 m g 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-phthalalde hyde derivatization method. The method was checked with several red wi nes from Tarragona by comparing the results with the standard pre-colu mn method.