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.     

The extrapolation of experimental equilibrium constant data to zero ionic strength: Critical review and new approach

Different Debye-Hückel expressions for the activity coefficients of species in aqueous solution in the ionic strength range I = 0-3.5m (3M) are used for the extrapolation of equilibrium constants data to I = 0 and the interpolation to unknown I values. This may be accomplished using four or more values of the equilibrium constants that are equally well distributed on the I scale. The interpolated and extrapolated equilibrium constant values obtained are quite satisfactory and within the experimental error of the corresponding equilibrium constants. The values at I 0.1 are very important as they can particularly influence the equilibrium constant value calculated at I = 0 and for which the error can reach 0.1 log unit or more. The values at I 1.5 can also influence the extrapolated value at I = 0 and the interpolated value at a given I when an inadequate extrapolation model is selected. Among the expressions used, only those with two or more unknown parameters are suitable for such calculations.