New quantitative determination of 2,3-dioxobutane (diacetyl) in alcoholic beverages by an indirect spectrofluorimetric procedure with use of 2,3-diaminonaphtalene as reagent of derivatization

The experimental conditions of an earlier spectrofluorimetric determination of 2,3-dioxobutane as 2,3-diaminonaphtalene derivative have been optimized. Fluorescence intensities are increased by a factor of 13 over the precedent ones. The calibration graphs are linear in the concentration range 2.5-130 mumol/l in spectrophotometry and 1.75 x 10(-1)-2.5 mumol/l in fluorimetry. Limits of detection are very small: 7 x 10(-1) and 2.5 x 10(-2) mumol/l, respectively. Applications to alcoholic beverages are described. The new fluorimetric technique is simple and rapid since it does not require steam distillation.     

Bioelectrochemical determination of lactic and malic acids in wine

Electroanalytical methods based on biosensor technology and differential pH measurement for the determination of l-lactic and l-malic acids in wine were developed. Lactic acid was determined according to two procedures consisting of the use of two lactate electrochemical biosensors, one based on a Clark O(2) probe and the other based on the H(2)O(2) probe. The two probes were assembled employing a polymeric membrane where the enzyme lactate oxidase was previously immobilized. Both sensors exhibited high storage and operational stabilities, and good reproducibility when used in wine matrix. Measurement of lactate was carried out using two different flow-through cells assembled with the lactate probes. The low detection limit of these probes and the relatively high concentration of lactate in wine samples required a serial dilution in the range 1:100 to 1:200, thus eliminating all potential electrochemical or enzyme interferences present in the sample. Malic acid was determined using a differential commercially available pH-meter and an enzymatic procedure. For this analysis, we employed the malic enzyme (EC 1.1.1.40) and the NADP(+) cofactor. The pH variation due to the action of this enzyme on malic acid was found to be proportional to the malic acid present in the sample. Experimental parameters such as pH, temperature, and co-factor concentration were optimized, resulting in malate determination in less than 1 min with good reproducibility. Various samples of wine and wine musts were assayed for lactate with the two biosensor procedures, and for malate with differential pH-metry. The results when compared with those obtained with the commonly used spectrophotometric procedure correlated well.