An amperometric biosensor for polyphenolic compounds in red wine.

In the present work, a biosensor was developed with Laccase Coriolus Versicolor as the biological reconnaissance element immobilized on derivatized polyethersulphone membranes and applied to a Pt-Ag, AgCl US electrode base. Its application to several polyphenols usually found in red wine (caffeic acid, gallic acid, catechin, rutin, trans-resveratrol, quercetin and malvidin) was tested. It was observed that an amperometric response was obtained for catechin at +100 mV (versus Ag, AgCl) and caffeic acid at -50 mV in acetate buffer solutions (pH 4.5) having 12% ethanol. At pH 3.5 and +100 mV the biosensor was sensitive to both substrates and their response was additive. A limit of detection of 1.0 x 10(-6) M, linearity ranging from 2.0 to 14.0 x 10(-6) M, high sensitivity (0.0566 mAM(-1)) and reproducibility (R.S.D. <10%) were achieved for equimolar mixed solutions of catechin and caffeic acid. Under the same experimental conditions the other polyphenols tested individually did not yield any biosensor response. The application of the biosensor to red wine samples required a previous solid phase extraction for polyphenols enrichment. In fact, attempts to apply the biosensor in red wine using the "standard addition" methodology showed that large interferences occurred, as was to be expected. Reduction currents of -0.33 +/- 0.03 nA were obtained when the biosensor was used with the wine extract at +100 mV. This current could be ascribed to catechin and caffeic acid, although some interference by other polyphenols at the matrix level seemed to persist. The present biosensor showed promising applications for the wine analysis in future.