Chinese Medicine Littoralis Polysaccharide Monosaccharide Composition And Determination Studies

This paper is about the content of polysaccharide in Radix Glehniae and the contents of monosaccharide compositions in polysaccharide .Phenol-sulfuric acid method was used for determination of the amount of total GLP. HPLC and GC were used for determination of contents of monosaccharide compositions in polysaccharide.Hot water extracting and ethanol precipitating method were employed to isolate polysaccharide. Protein was removed by Sevag method. The amount of total GLP was measured with phenol-sulfuric acid method. UV-spectrophotometer was used to determine the characteristic absorption and concentration of GLP. The content of GLP was good linearity over the range of 1.96⁶.86μg/mL (r = 0.9990) . The average recovery was 97.6%( RSD =1.5%).GLP sample was hydrolyzed with trifluoroacetic acid. Hydroxylamine hydrochloride, pyridine and acetic anhydride were added as derivative agents. The hydrolyzed monodaccharides were converted into corresponding aldoononitrile zcetates. The derivatives were determined by GC. DM-5 column (30 m×0. 25 mm, 0. 25μm) was used. The inlet temperature was maintained at 270℃. The column oven was begun at 160℃, then programmed from 160℃to 195℃at 2℃/min, hold for 6 min ,then programmed from 195℃to 230℃at 10℃/min, and finally, hold for 3 min. Helium at a constant flow rate of 30 mL/min was used as carrier gas. Concentrations of seven monosaccharides mannose, rhamnose, glucose, galactose, arabinose, and two unidentifiable monosaccharides were determined in GLP sample. The contents of mannose rhamnose, galacturonic-acid, glucose, galactose and arabinose were good linearity over the range of 2.4-24μg/mL(r = 0.9992), 6-60μg/mL (r = 0.9996), 1.2-12μg/mL (r = 0.9991), 140¹400μg/mL (r = 0.9991), 6-60μg/mL (r = 0.9994). The average recoveries were 91.0%( RSD =2.2%), 91.2%(RSD=2.2%), 87.3%(RSD=3.1%), 94.7%( RSD =2.3%), 90.5%( RSD =2.9%).GLP sample was hydrolyzed with trifluoroacetic acid.1-pheny-3-methyl-5-pyrazolone (PMP) was added as derivative agent. The hydrolyzed monodaccharides were converted into derivatives. The derivatives were determined by HPLC. The analytical column was C₁₈ (250 mm×4.6 mm, 5μm).The mobile phase was 19% (v/v) acetonitrle- ammoniumacetact (ammoniumacetact - glacial acetic acid pH 5.5), isocratic elution prosess. The flow rate was 1.0 mL /min and the column temperature was 35℃. The detector was ultriolet detector. The detection wavelength was 250 nm. Concentrations of eight monosaccharides-mannose, rhamnose, galacturonic-acid, glucose, galactose, arabinose and two unidentifiable monosaccharides were determined in GLP sample. The contents of mannose rhamnose, galacturonic-acid, glucose, galactose and arabinose were good linearity over the range of 0.72-14.4μg/mL (r = 0.9990), 2.18-43.6μg/mL (r = 0.9990), 11.87-237.4μg/mL (r = 0.9994), 304.2-6084μg/mL (r = 0.9990), 14.4-288μg/mL (r = 0.9990), 9.6-192μg/mL (r = 0.9992). The average recoveries were94.0%( RSD =2.5%), 88.5%( RSD =2.5%), 93.9%( RSD =2.3%), 95.9%( RSD =2.7%), 89.2%( RSD =2.5%)å'ŒP 90.8%( RSD =2.8%).