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Water-soluble polysaccharide from Gynostemma pentaphyllum herb tea (PSGP) was isolated by hot-water extraction and ethanol precipitation. The chemical components and preliminary immunomodulating activity of PSGP were investigated both in vitro and in vivo. Capillary zone electrophoresis analysis showed that PSGP was a typical nonstarch heteropolysaccharide, with glucose being the main component monosaccharide (23.2%), followed by galactose (18.9%), arabinose (10.5%), rhamnose (7.7%), galacturonic acid (4.7%), xylose (3.9%), mannose (3.1%), and glucuronic acid (1.2%). PSGP could significantly stimulate peritoneal macrophages to release nitric oxide, reactive oxygen species, and tumor necrosis factor-alpha in a dose-dependent manner. This immunostimulating activity of PSGP was further demonstrated by its inhibition on the proliferation of human colon carcinoma HT-29 and SW-116 cells incubated with the supernatant of PSGP-stimulated macrophage culture. It is evident that PSGP is a very important ingredient responsible for at least in part the immunomodulating activity of G. pentaphyllum herb tea.

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A simple capillary electrophoretic method based on 1-phenyl-3-methyl-5-pyrazolon (PMP) derivatization has been developed for simultaneous separation and determination of nine aldoses (xylose, arabinose, glucose, rhamnose, fucose, galactose, mannose, glucuronic acid and galacturonic acid). The separation of PMP-labeled monosaccharide derivatives was carried out in uncoated capillary (48.5 cm؋75 m mm i.d.) and under the selected optimum conditions of pH 11.0, 200 mM borate buffer at applied voltage 15 kV and capillary temperature 20°C, the nine PMP-monosaccharides could be perfectly separated from each other within 40 min. Furthermore, the developed method was firstly applied to determine the sugar composition in the polysaccharide isolated from Chinese Codonopsis pilosula. The results showed that C. pilosula polysaccharide was a typically acidic heteropolysaccharide and was composed of arabinose, glucose, rhamnose, galactose, mannose, glucuronic acid and galacturonic acid in the molar contents of 48.1, 103.5, 16.1, 48.5, 7.5, 4.2 and 119.1 m mM, respectively. The assay results were satisfactory. Key words Codonopsis pilosula; heteropolysaccharide; composition analysis; capillary zone electrophoresisBiol. Pharm. Bull. 31(10) 1860-1865 (2008) © 2008 Pharmaceutical Society of Japan * To whom correspondence should be addressed. e-mail: xbyang@snnu.edu.cn MATERIALS AND METHODS Materials and ReagentsThe plant materials of the roots of C. pilosula were harvested from the region of Qinling Mountains, Shaanxi province, China and were identified according to the identification standard of Pharmacopeia of the People's Republic of China. The plant materials were thoroughly air-dried and finely powdered.1-Phenyl-3-methyl-5-pyrazolone (PMP), purchased from Beijing Reagent Plant (Beijing, China), was re-crystallized twice from chromatographic grade methanol before use. DMannose, L-rhamnose, D-glucuronic acid, D-galacturonic acid, D-glucose, D-xylose, D-galactose, L-arabinose, D-fucose were all obtained from Sigma (St. Louis, U.S.A.). Trifluoroacetic acid (TFA) was obtained from Merck (Darmstadt, Germany). HPLC grade acetonitrile and methanol were purchased from TEDIA (U.S.A.). Boric acid and sodium hydroxide were obtained from Sigma (U.S.A.). HPLC grade methanol (MeOH) was from TEDIA (U.S.A.). All aqueous solutions were made up in the Milli-Q system water unless mentioned otherwise. All the other chemicals were of analytical grade.Isolation of C. Pilosula Polysaccharide The C. pilosula polysaccharide was isolated by hot-water extraction and ethanol precipitation. The dried materials of Chinese C. pilosula (550 g) were defatted with 95% alcohol and then extracted with distilled water (g/mlϭ1 : 10) for 3 h. After each three-hour period of water extracting, the water extracts were collected and the residues were extracted again for 3 cycles. The combined extracts were pooled, concentrated to 30% of the original volume under a reduced pressure and then centrifuged at 3000 rpm for 15 min. The supernatant was collected and 3 volum...