天然产物研究与开发 ›› 2017, Vol. 29 ›› Issue (7): 1148-1153.doi: 10.16333/j.1001-6880.2017.7.012

• 研究简报 • 上一篇    下一篇

金花茶叶多糖的分离纯化及组成分析

许子竞*,石谦,罗树常   

  1. 贵州工程应用技术学院天然产物中心,毕节 551700
  • 出版日期:2017-07-29 发布日期:2017-08-11

Extraction,Purification and Component Analysis of Polysaccharide from Camellia chrysantha (Hu) Tuyama Leaves

XU Zi-Jing*,SHI Qian,LUO Shu-Chang   

  1. Center of Natural Products,Guizhou University of Engineering Science,Bijie 551700,China
  • Online:2017-07-29 Published:2017-08-11

摘要: 以金花茶叶为原料,以纯化水为溶剂提取金花茶叶粗多糖(Crude polysaccharides from dry leaves of Blumea riparia,CCTLP),经过D101大孔树脂脱色,上DEAE-纤维素柱分离,依次用水、0.15 mol/L NaCl、0.3 mol/L NaCl、0.45 mol/L NaCl溶液梯度洗脱,得CCTLP-0、CCTLP-0.15、CCTLP-0.3和CCTLP-0.45四个洗脱部分;将CCTLP-0、CCTLP-0.15部分采用Sevage法脱蛋白、活水透析、Sephadex G-15柱层析脱盐,再经琼脂糖Sepharose 6FF柱层析分离纯化,得到CCTLP-0-1、CCTLP-0.15-1、CCTLP-0.15-2、CCTLP-0.15-3和CCTLP-0.15-4五个组份;经GPC检测其分子量及分子量分布和GPC峰型表明,确定它们为均一组份多糖;CCTLP-0、CCTLP-0.15酸解,经HPLC分析其组份结果显示,CCTLP-0和CCTLP-0.15洗脱部分主要由鼠李糖、果糖和半乳糖组成,其摩尔质量比大致为:1.00:1.066:0.384和1.00:1.186:0.544。

关键词: 金花茶叶多糖, DEAE-纤维素分离, 凝胶色谱纯化, GPC检测, 多糖分析

Abstract: Crude polysaccharides (CCTLP) were extracted from dry leaves of Camellia chrysantha (Hu)Tuyama using water.The four parts including CCTLP-0,CCTLP-0.15,CCTLP-0.3 and CCTLP-0.45 were obtained from CCTLP through sequentially gradient elution by water,0.15 mol/L NaCl,0.3 mol/L NaCl and 0.45 mol/L NaCl as elution reagent by DEAE-cellulose column after removing pigments of CCTLP by D101 macroporous resin.CCTLP-0 and CCTLP-0.15 were then deproteinized by Sevage method,running water dialysis and removing residual NaCl by Sephadex G-15 chromatography for further purification.The five fractions including CCTLP-0-1,CCTLP-0.15-1,CCTLP-0.15-2,CCTLP-0.15-3 and CCTLP-0.15-0.4 were obtained by purification from CCTLP-0 and CCTLP-0.15 using agarose Sepharose 6FF column chromatography.The results showed that they were homogeneous polysaccharide through GPC detection in molecular weight and molecular weight distribution and in analysis of GPC peak type detected.Both CCTLP-0 and CCTLP-0.15 were mainly composted of rhamnose,fructose and galactose through HPLC analysis after they were acid hydrolysis,which molar mass ratios were similar to 1.00:1.066:0.384 and 1.00:1.186:0.544,respectively.

Key words: polysachaide from Camellia chrysantha (Hu) Tuyama leaves, DEAE-cellulose column separation, gel permeation chromatography purification, GPC detection, polysaccharide analysis