Monitoring of Chlorogenic Acid and Antioxidant Capacity of Solanum melongena L. (Eggplant) under Different Heat and Storage Treatments.

Solanum melongena L., also known as eggplant, is a widely consumed vegetable and it is well-known for its beneficial antioxidant properties, due to phenolic compounds. In this work, the influence of different cooking procedures on the content of chlorogenic acid was evaluated on eggplant samples of different geographic origin by high-performance liquid chromatography (HPLC). An easy and quick extraction procedure with 50% methanol as the extraction solvent was optimized for the first time by means of a design-of-experiment and applied to heat treated samples of eggplant. The antioxidant capacity of eggplant extracts was also evaluated by using the ABTS assay and it was correlated with the data obtained by the HPLC method. The content of chlorogenic acid was different in each heat-treated eggplant sample and it depended on the temperature applied during the cooking procedure. In particular, an increase of chlorogenic acid content with rising temperature was observed. Conversely, a very high temperature (250 °C) caused a decrease of chlorogenic acid amount. The influence of storage on the content of chlorogenic acid was also monitored. While the level of chlorogenic acid in fresh samples decreased during four weeks of storage, an increase in its content in heat treated eggplant was observed within the same period. Multivariate data analysis was used to classify eggplant samples into different groups, according to the country of origin and heat treatment procedure. This study provides new insights to preserve the antioxidant properties of eggplant phenolics during different thermal and storage treatments in order to highlight their health promoting effects.

Flow injection tyrosinase biosensor for direct determination of acetaminophen in human urine.

An amperometric biosensor compatible with a flow injection analysis (FIA) for highly selective determination of acetaminophen (APAP) in a sample of human urine was developed. This biosensor is also suitable for use in the routine pharmaceutical practice. To prove this statement, two different commercially available pharmaceutical formulations were analyzed. This nano-(bio)electroanalytical device was made from a commercially available screen-printed carbon electrode covered by a thin layer of non-functionalized graphene (NFG) as amperometric transducer. A biorecognition layer was prepared from mushroom (Agaricus bisporus) tyrosinase (EC 1.14.18.1) cross-linked using glutaraldehyde, where resulting aggregates were covered by Nafion®, a known ion exchange membrane. Owing to the use of tyrosinase and presence of NFG, the developed analytical instrument is able to measure even at potentials of 0 V. Linear ranges differ according to choice of detection potential, namely up to 130 µmol L-1 at 0 V, up to 90 µmol L-1 at -0.1 V, and up to 70 µmol L-1 at -0.15 V. The first mentioned linear range is described by the equation Ip [µA] = 0.236 - 0.1984c [µmol L-1] and correlation coefficient r = 0.9987; this equation was used to quantify the content of APAP in each sample. The limit of detection of APAP was estimated to be 1.1 µmol L-1. A recovery of 96.8% (c = 25 µmol L-1, n = 5 measurements) was calculated. The obtained results show that FIA is a very selective method for APAP determination, being comparable to the chosen reference method of reversed-phase high-performance liquid chromatography.

Development of extraction method for characterization of free and bonded polyphenols in barley (Hordeum vulgare L.) grown in Czech Republic using liquid chromatography-tandem mass spectrometry.

Complete characterizations of free and bonded phenolic compounds, presented in four cultivars of barley from two regions of Czech Republic, were achieved, using optimized solvent extraction and liquid chromatography coupled with tandem mass spectrometry. The optimization of extraction of free polyphenols was performed using Box-Behnken design and response surface methodology. The intra-day and extra-day precision of developed method were below 6% and 12%, respectively. The isolation of polyphenols bonded to the cell wall structure was carried out by a hydrolysis process. In all cultivars, p-hydroxybenzoic, p-coumaric and ferulic acids were the most abundant compounds. Their average amounts in barley samples were 17.6, 15.2 and 54.4% (m/m), respectively. The highest amount of these compounds was found in the bonded form, proving the importance of this procedure for the correct characterization of total polyphenols in food matrices.

Fast gradient HPLC/MS separation of phenolics in green tea to monitor their degradation.

The degradation of catechins and other phenolics in green tea infusions were monitored using fast HPLC/MS separation. The final separation was performed within 2.5min using Ascentis Express C18 column (50mm×2.1mm i.d.) packed with 2µm porous shell particles. Degradation was studied in relation to the temperature of water (70, 80, 90°C) and the standing time of the infusion (up to 6h). Along with chromatographic separation, the antioxidant properties of the infusions were monitored using two spectrophotometric methods. During staying of green tea infusion, the degradation of some catechins probably to gallic acid was observed. Finally, the influence of tea bag storage on antioxidant properties of green tea was evaluated. Rapid degradation of antioxidants after 3weeks was observed. The principal component analysis, factor analysis and discriminant analysis were used for the statistical evaluation of obtained experimental data.