, 2009). Firstly, β-carotene (0.2 mg) was dissolved in 1.0 mL chloroform. After, 0.02 mL linoleic acid plus 0.2 mL Tween 80 were added and the mixture was left standing at room temperature for 15 min. Anticancer Compound Library clinical trial After evaporation of chloroform, 50 mL of oxygenated distilled water was added and the mixture was shaken to form an emulsion (β-carotene–linoleic acid emulsion). Aliquots of 3.0 mL of this emulsion were transferred into test tubes containing 0.2 mL of different concentrations of extracts. The tubes were shaken and incubated at 50 °C

in a water bath. As soon as the emulsion was added to each tube, the zero time absorbance (A0) was measured at 470 nm. A second absorbance (A1) was measured after 120 min. A blank, without http://www.selleckchem.com/products/Rapamycin.html β-carotene was prepared for back-ground subtraction. Lipid peroxidation (LPO) inhibition was calculated using the following equation: LPO inhibition(%)=A0−A1A0×100. The assays were carried out in triplicate and the results expressed as mean

values ± standard deviations. The extract concentration producing 50% antioxidant activity (EC50) was calculated from the graph of antioxidant activity percentage against the extract concentration. Gallic acid, syringic acid and pyrogallol were used as standards. DPPH, ABTS, potassium persulfate, β-carotene, linoleic acid, phenolic acids, flavonoids, aromatic compounds, organic acids, Folin–Ciocalteu’s phenol reagent and were obtained from Sigma Chemical Co. All other chemicals were of analytical grade. All analyses were performed in triplicate. The data were expressed as means ± standard deviations and one-way analysis of variance (ANOVA) and Tukey test were carried out to assess for any significant differences between the means. Differences between means at the 5% (P < 0.05)

level were considered significant. Fig. 1 shows the curve of growth Grape seed extract of A. brasiliensis in submerged cultures. Maximum production of biomass (10.2 ± 1.10 g/L) was obtained after 4 days of cultivation in the beginning of stationary growth phase. After that, the analysis of residual reducing sugars showed depletion of glucose and a decline in dry weight owed to autolysis of the fungi (late stationary growth phase). To evaluate the main chemical components as well as the antioxidant properties, mycelia obtained at two times of cultivation were collected, one after 4 days of cultivation (designated in this work as young mycelia) and another after 8 days (here designated as old mycelia). High extraction yields were obtained from three materials using ethanol:water (70:30): 42.5 ± 1.4 g/100 g, 48.3 ± 1.8 g/100 g and 44.9 ± 1.2 g/100 g, for A. brasiliensis fruiting bodies, young mycelium and old mycelium, respectively. Table 1 shows the chemical characterization of the A. brasiliensis hydroalcoholic extracts obtained from three materials. The extracts presented high amounts of carbohydrates, mostly of the non-reducing type.