Determination of Biogenic Amines in Wine After Clean-Up by Solid-Phase Extraction O. Busto *1 / M. Mestres 1 / J. Guasch 1 / F. Borrull 2 1Departament de Qufmica, Escola d'Enologia, Universitat Rovira i Virgili, Ram6n y Cajal, 70, 43005 Tarragona, Spain 2Departament de Qufmica, Facultat de Qufmica, Universitat Rovira i Virgili, Pqa. Imperial Tarraco, 1, 43005 Tarragona, Spain Key Words Column liquid chromatography Solid-phase extraction Wine Biogenic amines Phthalaldehyde Summary A suitable method for the determination of 16 biogenic amines in wine has been developed. The method involves dean-up of wine samples using ion-exchange cartridges and a preconcentration step, under control- led vacuum, before derivatization of the amines by treatment with phthalaldehyde (PA) and reversed- phase HPLC with gradient elution and fluorimetric detection. Linearity of response was obtained for all the biogenic amines from 100 pg L- 1 to 10 mg L- 1. Limits of detection for the amines were similar for all PA-derivatives (25-50 ggL -1) and the quantitation limits were about 0.1mgL -1. After clean-up and preconcentration, the concentration levels increased 10-fold for all amines except putrescine and cadaverine, which gave poor recovery by this method unlike the rest which gave recoveries of almost 90 %. The overall process was successfully applied to identify and quanti- fy biogenic amines in several red wines from the Tarragona region. Introduction It has been known since 1954 that biogenic amines are naturally occurring compounds in various food prod- ucts, such as fish, cheese, wine and fermented sausages [1, 2]. There is a great diversity of these amines in food and beverages [3-6], their content being studied because of their direct and indirect potential toxicity. The interac- tion between ethanol (a monoamine oxidase inhibitor) and amines seems to be synergistic, which is important for those wine consumers who are sensitive to such compounds. Among other biogenic amines, tyramine, isoamylamine, tryptamine and 2-phenethylamine produce hyperten- sive effects; cadaverine and putrescine enhance the anaphylactic activity and toxicity of histamine by simultaneous oral administration [7]. It is still not fully known how other amines interact with each other. All analytical methods employed for biogenic amines involve two steps: isolation and determination. In the former, problems may appear arising from their differ- ent chemical structures. In that case it is necessary to find a middle course in order to obtain adequate recovery for each amine. The determination and concentration of amines in wine have been reported by several investigators: Subden [7], Vidal [8], Cerutti [5], Baucom [9], Zappavigna [10] and Fr61ich [11], whose work includes a step to increase the selectivity thus lowering the detection limit of the methods. As can be imagined, direct determination of these compounds in wine is difficult due to their low concentration and the complexity of the wine matrix. Hence it is necessary prior to determining concentra- tion levels of biogenic amines in wine, to undertake clean-up and preconcentration steps. In order to isolate compounds of interest, liquid-liquid extraction (LLE) was investigated using different mixtures of organic solvents prior to derivatization of the sample and therefore avoiding troubles with selec- tivity. Some investigators carry out this extraction procedure after the derivatization process, improving selectivity but not recovery. As indicated in a previous paper [12], these LLE procedures often result in very low analytical recoveries. Furthermore, when PA- derivatives are synthesized it is necessary to take into account their stability. A highly promising approach is preconcentration of the compounds of interest on suitable adsorbents, prior to their separation and detection by a chromatographic 404 0009-5893/95/04 0404-07 $ 3.00/0 Chromatographia Vol. 40, No. 7/8, April 1995 9 1995 Friedr. Vieweg & Sohn Verlagsgesellschaft mbH Original