Then 10 μl of hydrogen peroxide (H2O2) as oxidant was added in each tube. Growth of the yeast culture was monitored taking absorbance at 600 nm at the end of 20 h. The effect of phenolic extracts in presence of oxidants on the net growth of yeast cells was determined according to the following equation. Ayeast growth=Atest sample−AcontrolAcontrol×100Where Ayeastgrowth = net growth of H2O2 induced yeast cells after treatment with phenolic extracts, Acontrol = absorbance of yeast cells in presence of H2O2, Atestsample = absorbance

of yeast cells in presence of H2O2 and phenolic extracts. Water extracts (50 ml) of unfermented www.selleckchem.com/products/nu7441.html and fermented wheat were extracted with ethyl acetate [1:1; v/v] for 30 min in a separating funnel. The ethyl acetate fractions were evaporated to dryness and reconstituted in methanol. Now the phenolic extract

was filtered through 0.45 μm Supor®-450 membrane disc filter (Pall Gelman Laboratory, USA) and then thin layer chromatography (TLC) of PCs was performed on silica gel plates using mobile phase chloroform:methanol:formic acid [85:15:1; v/v/v] and visualized under short wave (254 nm) and long wave UV light (365 nm). Same samples (2 μl) were analyzed by an ultra-performance liquid chromatography (Waters, Milford, USA). The separation of phenolics was performed on a BEH 300C-18 column (2.1 mm × 50 mm, 1.7 μm). The column temperature, total run time and flow rate were set at 30 °C, 5 min and 0.6 ml/min, respectively. Two mobile phases consisted of buy PLX-4720 NADPH-cytochrome-c2 reductase water containing 0.1% TFA (solvent A) and acetonitrile containing 0.1% TFA (solvent B) were used and gradient elution was carried out using the following program: 95% A to 90% A in 1 min, 90% A to 85% A in 1 min, 85% A to 75% A in 1 min, 75% A to 40% A in 1 min, 40% A to 0% A in 0.2 min, 0% A to 0% A in 0.6 min and 0% A to 95% A in 0.2 min. The peaks were identified

by congruent retention times and UV spectra (280 nm) and compared with standards and quantified based on their peak’s area. The mean values and the standard deviations were calculated from the data obtained from experiments in triplicates. The data were analyzed by one-way analysis of variance (ANOVA). It is well known from literature data that extraction conditions and characteristics of the sample can affect the efficiency of the extraction, independently or interactively. The solvent and the temperature are the process parameters that usually have the greatest impact on the efficiency of extraction of bioactive compounds from the plant material. In general, alcohol/water solutions exert a better influence on the extractability of phenolic compounds in comparison to the mono-component solvents.