In addition, the in vitro antioxidant activities of the pectic fraction and a methanolic extract were investigated. 1,1-Diphenyl-2-picryldydrazyl (DPPH ), thiobarbituric acid, butylated hydroxyanisole (BHA), 3-phenylphenol, ethylenediaminetetraacetic acid (EDTA), DEAE-Trisacryl® Plus, N-cyclohexyl-N′-(2-morpholinoethyl)carbodiimide, metho-p-toluenesulfonate, sodium borodeuteride, ascorbic acid, bovine serum albumin (BSA), deoxyribose,
l-arabinose, d-xylose, d-glucose, d-mannose, d-galactose, l-fucose, l-rhamnose, Transmembrane Transporters modulator d-galacturonic acid and dextran standards were purchased from Sigma–Aldrich Chemical Co. (St. Louis, MO, USA)., Methyl iodide (MeI), phenol, hydrogen peroxide (H2O2) and sulphuric acid were from Merck Co. (Darmstadt, Germany). All other chemicals used were of analytical grade. A commercial sample of guarana (P. cupana) seed powder (batch number 8656A8) was kindly supplied by Herbarium Laboratório Botânico (Paraná, Brazil), which is a herbal medicine pharmaceutical laboratory with a line of products made up of herbal medicine and nutritional supplements. To prepare the guarana AZD6244 cost powder, whole dried seeds of P. cupana L.) Kuntze, collected in Bahia-Brazil, were ground in a hammer mill (60 mesh
sieve-95%). The guarana powder was defatted with toluene:ethanol (2:1, v/v) in a Soxhlet extractor (48 h). Subsequently, the dried material was treated with methanol:water (4:1, v/v) under reflux for 20 min and was immediately cooled to room temperature and centrifuged. This residue (residue 1) was dried and used for polysaccharide extraction as follows. Residue 1 was used Akt inhibitor for polysaccharide extraction according to the scheme depicted in Fig. 1. The extraction procedure was based on the work of Bochicchio, Petkowicz, Alquini, Busato,
and Reicher (2006) with some modifications. Successive extractions were performed in a mechanical blender. After each extraction, the sample was centrifuged, and the residue was subjected to the next extraction step. Each extract was concentrated and treated with ethanol (2:1 v/v) to obtain the precipitated polysaccharides, which were then washed three times with ethanol and dried under a vacuum. The residue 1 was first extracted with DMSO (2×) at 25 °C for 24 h and 120 h to produce fractions GD-I and GD-II, respectively. Residue 2 was subjected to sequential aqueous extractions at 25 °C (2×) and at 90 °C (2×) for 4 h each. Four fractions were obtained: GW-I (25 °C), GW-II (25 °C), GHW-I (90 °C), and GHW-II (90 °C). Then, alkaline extractions with 2 M (2×) then 4 M NaOH (2×) were performed at 25 °C for 120 min in the presence of NaBH4. Each extract was neutralised with aqueous 50% acetic acid, and the precipitated polysaccharides (hemicellulose A) were isolated by centrifugation. The resulting supernatants were dialysed, concentrated to a small volume and then precipitated with ethanol (2:1 v/v) to yield hemicellulose B fractions.