黑果枸杞多糖的纯化工艺研究

Study on the Purification Technology of Lycium ruthenicum Polysaccharides

研究黑果枸杞(Lycium ruthenicum Murr)水溶性多糖的纯化工艺。结果表明,三氯乙酸法为最佳脱蛋白方法,过氧化氢法为最佳脱色方法,脱色条件为温度45℃,时间30min,pH 8～9,用量为每100mL粗糖液5mL H2O2。黑果枸杞经超声加热提取、醇沉、脱蛋白、脱色后得黑果枸杞粗多糖(crude Lycium rutheni-cum polysaccharides,CLRP)。CLRP经DEAE-Cellulose柱层析,用蒸馏水以及0.05、0.1、0.25、0.5mol/LNaHCO3洗脱液进行分段洗脱,分离得到LRP1、LRP2、LRP3、LRP4、LRP5五个多糖组分,多糖含量最高的LRP5经Sephadex G-100柱层析后得到LRGP5。经高效凝胶渗透色谱法分析,LRGP5为均一多糖,测得相对分子质量约为1.37×105。

The purification technology of water-soluble polysaccharides from Lycium ruthenicum Murr was studied.TCA method was optimal for the deproteinization effect.The process of depigmentation of H2O2 was： adding 5 mL H2O2 per 100 mL crude polysaccharide solution and depigmentating for 30 min at 45 ℃ and pH 8-9.Crude Lycium ruthenicum polysaccharides（CLRP） were isolated through ultrasonic heating extraction,ethanol precipitation,deproteinazation and depigmentation.CLRP could be separated by DEAE-cellulose column chromatography,and gradiantly eluted with distilled water,0.05,0.1,0.25,0.5 mol/L NaHCO3 eluant to get five components named LRP1,LRP2,LRP3,LRP4 and LRP5.Then LRP5 was further purified by Sephadex G-100 column chromatography,and a sub-fraction was obtained and it was named LRGP5.High performance gel permeation chromatography（HPGPC） showed that the LRGP5 was homogeneous polysaccharide with an average relative molecular mass of 1.37×105.