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    高效阴离子交换色谱-脉冲安培检测法分析漏芦多糖中单糖组成

    Determination of Monosaccharides in Polysaccharides of Rhaponticum Uniforum by High-performance Anion Exchange Chromatography with Amperometric Detection

    • 摘要: 采用高效阴离子交换色谱-脉冲安培检测法(HPAEC-PAD),建立了一种测定漏芦多糖中单糖组成的方法.水提、醇沉法提取粗多糖,Sevage法除蛋白,用2 mol·L-1硫酸在100 ℃水解多糖,以CarboPac PA10色谱柱(2 mm×250 mm)为分离柱,100 mmol·L-1氢氧化钠溶液为淋洗液,流速为1.0 mL·min-1,脉冲安培检测,总分析时间为6 min.测定漏芦多糖中单糖组成为阿拉伯糖、葡萄糖和果糖,其摩尔比为1.55∶1.00∶2.10;加标回收率为93.0%~107.8%,检出限为2.8~12.3 μg·L-1.

       

      Abstract: High performance anion-exchange chromatography with pulse-amperometric detection (HPAEC-PAD) was applied to the determination of monosaccharides in polysaccharide of Rhaponticum uniforum.Polysaccharide was separated from the sample by refluxing-extraction with C2H5OH-H2O (80+20) for 3 h to defat.After volatilization of ethanol,the drug residue was extracted twice with boiling water,maintaining 1 h for each extraction.Ethanol (95%) was added to the water extract in a volume of 200 mL to make content of ethanol in the solution achieved to 4/5 of its total volume.Polysaccharides in the ethanol solution were precipitated overnight at 4 ℃.Protein in the crude polysaccharide was removed by the Sevage method.One mg of the polysaccharide was hydrolyzed for 4 h at 100 ℃ in 2 mol·L-1 H2SO4,and then separated on the CarboPac PA10 chromatographic column.100 mmol·L-1 NaOH solution was used as eluant passing through the column at a flow-rate of 1.0 mL·min-1.Monosaccharides in the eluate were determined by PAD.Six min were taken for the whole analysis.It was found that arabinnose,glucose and fructose in the mole ratio of 1.55 to 1.00 to 2.10 were the 3 monosaccharides in polysaccharide of Rhaponticum uniforum.Detection limits found were in the range from 2.8 μg·L-1 to 12.3 μg·L-1,and recoveries found by standard addition were in the range of 93.0%-107.8%

       

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