A simple phosphorus determination in walnuts and assessment of the assimilable fraction.

In this work, a method for the determination of phosphorus in walnuts has been developed due to the fact that there is no specific method for this purpose. A closed-vessel microwave-assisted digestion of the samples was optimized and a modified molybdenum blue spectrophotometric method was carried out with excellent precision and selectivity. To differentiate assimilable and non-assimilable phosphorus, an easy extraction procedure using 1 M NH4Cl was used. FTIR spectra ensured that the phytate extraction was successful. A sampling of thirteen different walnuts available to consumers from different crops and geographical origin was performed. Comparative statistical tools to differentiate walnuts were also carried out. Significant differences based on total content of phosphorus were obtained. Finally, a correlation study between total phosphorus concentration and the non-assimilable fraction showed a non-linear relationship (R2 = 0.435). Therefore, in relation to consumer health the total phosphorus determination must be complemented with information about assimilable phosphorus.

Selective methods for polyphenols and sulphur dioxide determination in wines.

A critical review to the methods recommended by international bodies and widely used in the winery industry and research studies was performed. A Laccase biosensor was applied to the selective determination of polyphenols in wines. The biosensor response was characterised and it responds mainly to o-diphenols which are the principal polyphenols responsible for the stability and sensory qualities of wines. The spectrophotometric method to determine free and total sulphur dioxide recommended for beers was applied directly to wines. A sampling of 14 red and white wines was performed and they were analysed for biosensor polyphenol index (IBP) and sulphur dioxide concentration (SO2). The antioxidant capacity by the ABTS(+) spectrophotometric method was also determined. A correlation study was performed to elucidate the influence of the polyphenols and SO2 on the wines stability. High correlations were found between IBP and antioxidant capacity and low correlation between SO2 and antioxidant capacity. To evaluate the benefits of wine drinking a new parameter (IBP/SO2) is proposed.

A comparison of three amperometric phenoloxidase-Sonogel-Carbon based biosensors for determination of polyphenols in beers.

Three phenoloxidases based biosensors were successfully developed using as electrochemical transducer a new type of electrode recently developed by our group: the "Sonogel-Carbon electrode". The employed enzymes were Trametes versicolor laccase (Lac), Mushroom tyrosinase (Tyr), and Horseradish peroxidase (HRP). Immobilization step was accomplished by doping the electrode surface with a mixture of the individual enzyme and Nafion ion exchanger as additive-protective. The biosensor responses, optimized in beer real samples, were evaluated for five individual polyphenols. It was found that the developed biosensors were sensitive to nanomolar concentrations of the tested polyphenols. As example, the limit of detection, sensitivity, and response linear range for caffeic acid for Nafion-Lac/Sonogel-Carbon biosensor were 0.06µmolL(-1), 99.6nAµmol(-1)L, and 0.04-2µmolL(-1), respectively. The stability and reproducibility of the biosensors were evaluated by applying them directly to beer real samples. It has been demonstrated that the Nafion-Lac/Sonogel-Carbon system is the more stable with a relative standard deviation of 3.3% (n=10), maintaining 84% of its stable response for at least three weeks. Estimation of polyphenol index in eight lager beers and a comparison of the results with those obtained by a classical method was carried out.