DEAE-cellulose was purchased from Shanghai Chemical Reagent Study Institute (Shanghai, China). components of ginseng [1]. Many studies have been carried out on ginseng polysaccharides concerning their purification, structural analysis and bioactivities since 1966. Among these reports, most of the polysaccharides were extracted from ginseng with hot water [2C6]. In recent years, our laboratory offers carried out a series of studies about structure–activity associations of water-soluble ginseng polysaccharides. Our results indicated water-soluble ginseng polysaccharides contained starch-like glucans, homogalacturonan (HG), rhamnogalacturonan-I (RG-I) and arabinogalactan (AG)-type pectin [7]. These polysaccharides exhibited numerous activities including inhibitory effects against galectin-3 [8], immunomodulatory effects [9], inhibition of cell migration [10] and anti-fatigue activity [11]. Owing to the interference of starch granules, some of the polysaccharides could not become extracted from ginseng with hot water. We then performed an extraction by -amylase answer after hot water extraction, and polysaccharides with different chemical structures compared with water-soluble ginseng polysaccharides were obtained [12]. As some pectic polysaccharides usually bind Ca2+ ions within the lamella Acadesine (Aicar,NSC 105823) of cell walls, forming so-called egg-box constructions, they could not become extracted very easily with water only. However, these polysaccharides can be solubilized by using chelating agents. Consequently, we used EDTA answer to further draw out these polysaccharides from ginseng origins [13]. Although ginseng has already been extracted by different methods, there might be some non-cellulosic polysaccharides still present in ginseng origins. To get the content and structural info of these non-cellulosic polysaccharides, other extraction methods should be applied. Alkali solutions have been utilized for obtaining pectin and hemicellulose which are highly branched and cross-linked in cell walls [14]. Therefore, in this study, 50?mM Na2CO3, 1?M KOH and 4?M KOH solutions were used in sequence to extract Acadesine (Aicar,NSC 105823) polysaccharides from ginseng Acadesine (Aicar,NSC 105823) origins after hot water, -amylase and EDTA solution treatment. The structures of these alkali-soluble polysaccharides were analysed by NMR, monoclonal antibody and high performance anion exchange chromatography (HPAEC). Based on the results of present and earlier studies, we could possess a comprehensive understanding of the total polysaccharides in ginseng. 2.?Material and methods 2.1. Materials Ginseng was cultivated and collected from Changbai Mountain, Jilin Province, China. Sepharose CL-6B was purchased from GE Healthcare (Pittsburgh, USA). DEAE-cellulose was purchased from Shanghai Chemical Reagent Study Institute (Shanghai, China). -Amylase (E.C.3.2.1.1) from spp. was purchased from SigmaCAldrich (St Louis, MO, USA). Acadesine (Aicar,NSC 105823) Endo-1,4–glucanase (EC 3.2.1.4) from was purchased from Megazyme (Bray, Ireland). All other reagents were of analytical or high performance liquid chromatography (HPLC) grade. 2.2. General methods Total carbohydrate content material was determined by the phenol–sulfuric acid method, using a mixture of major monosaccharides as the Mouse monoclonal to INHA standard [15]. Uronic acid content was determined by the [23]. NMR results indicated that RG-I domains in NGP-1a, NGP-2a and NGP-2b might be highly branched with -1,4-galactan, -1,5/1,3,5-arabinan and type II arabinogalactan part chains. Earlier studies also showed that 50?mM Na2CO3 mainly solubilized ramified hairy region of pectic polysaccharides from squash cell walls [24] and [14]. Open in a separate window Number 3. The 13C NMR spectra of ((ppm)and 1,2,4-Rharesidues were highly branched in the O-4 position in these fractions, with degree of branching of Acadesine (Aicar,NSC 105823) 86.8%, 62.5% and 54.5%, respectively. The majority of Ara residues present in these fractions were in the form of 1,5-, 1,3,5- and t-Araresidues reflected a large number of galactan part chains in RG-I. The glycosyl linkages of 1 1,3-, 1,6-, and 1,3,6-Galwere indicative of the presence of type II arabinogalactan part chains [25]. These results were consistent with the analysis by NMR spectra. Table?4. The glycosidic linkage type and molar percentage based on methylation and GC-MS analysis. (mentioned G) of which part chains were attached to O-6 [29]. The main part chains.