1.12 ), TPS1-1 (0.1 M NaCl fraction, yield 1.82 ), TPS1-2 (0.2 M NaCl fraction, yield 12.86 ), TPS1-4 (0.four M NaCl fraction, yield four.46 ) and TPS1-20 (two.0 M NaCl fraction, yield three.88 ) (Figure 3A). The sugar content material was detected by the phenol-sulfuric acid process. The normalised phagocytosis with the sub-fractions of TPS1 are shown in Figure 2B. The results showed TPS1-0 TPS1-1 and TPS1-2 fractions have the similar phagocytic activity, but are larger than other sub-fractions. Contemplating each the superior activity and yield, TPS1-2 was further fractionated utilizing SephacrylTM S-300 higher resolution column (Figure 3B), to get carbohydrate fractions of TPS1-2a (yield 20.0 from TPS1-2) and TPS1-2b (yield 22.5 from TPS1-2). Both TPS1-2a and TPS1-2b have been homogeneous, as they have been eluted at a single symmetrical peak (the second peak was the NaCl peak) from high performance gel permeation chromatography (HPGPC) as shown in Figure 3C, (1) and (2), using a molecular weight of 22 and 20 kDa, respectively. Figure 1. Flow chart from the isolation of aqueous polysaccharides from green tea.Int. J. Mol. Sci. 2014, 15 Figure 2. Phagocytic activity of (A) TPS1, TPS2 (18.9 or 1.89 g/mL) and (B) sub-fractions of TPS1 (1.89 g/mL). Constructive manage (LPS, 1 g/mL).Figure 3. (A) Profile of TPS1 in DEAE-cellulose column (Tube volume: 15 mL); (B) TPS1-2 in High-Resolution SephacrylTM S-300 (TPS1-2a: 180?20 min and TPS1-2b: 240?80 min), and TPS1-2a and TPS1-2b in HPGPC (C (1), 0?5 min; C (two), 20?9 min, a small gap at Y-axis of NaCl peaks as the eluants of slightly various NaCl concentration have been applied to elute).Int. J. Mol. Sci. 2014, 15 two.2. Monosaccharide Evaluation and Degree of EsterificationComplete hydrolysis of TPS1-2a and TPS1-2b followed by TLC (thin layer chromatography) showed that each polysaccharides contained only uronic acid. This was confirmed by the m-hydroxybiphenyl strategy [23]. Each TPS1-2a and TPS1-2b had been found to include one hundred of uronic acid employing D-galacturonic acid as a normal. TPS1-2a and TPS1-2b have been lowered 3 instances with sodium borohydride by Taylor’s system [24] to give the carboxyl-reduced polysaccharides, TPS1-2are and TPS1-2bre. Comprehensive hydrolysis of TPS1-2are and TPS1-2bre followed by TLC analysis also indicated that they did not contain uronic acid. After the hydrolysates had been converted into their corresponding alditol acetates and analyzed by GC-MS, each TPS1-2are and TPS1-2bre, only galactose resulted from galacturonosyl residues present in TPS1-2a and TPS1-2b was located (Figure 4A). The configuration of galactose in TPS1-2are and TPS1-2bre (Figure 4B) were assigned because the D configuration by comparing them with D-galactose and L-galactose requirements utilizing the GC-MS method developed by Situations et al.2-(1H-Pyrazol-3-yl)propan-2-ol Chemscene [25].2089377-51-3 custom synthesis Therefore D-galacturonic acid will be the big uronic acid present in TPS1-2a and TPS1-2b with about 28.PMID:24377291 4 and 26.1 of carboxylic groups present as methyl ester in galacturonic acid residues, respectively. The degrees of O-acetylation at 2-O and/or 3-O were determined as only 0.48 and 0.79 for TPS1-2a and TPS1-2b. Figure four. (A) Monosaccharide composition of TPS1-2are and TPS1-2bre, reduced kind of TPS1-2a and TPS1-2b, respectively; (B) Configuration of TPS1-2are and TPS1-2bre comparing to D-Gal and L-Gal standards.Int. J. Mol. Sci. 2014, 15 Figure 4. Cont.two.3. Periodate Oxidation In accordance with the results of periodate oxidation, 1 mol of GalA residues in TPS1-2a and TPS1-2b consumed 0.97 and 0.98 mol NaIO4, respectively, in approximate accor.