采用DEAE-纤维素柱层析法对海蚬粗多糖(CP)进行初步分离,获得3个组分(F1、F2、F3)。采用Sephdex G-100凝胶柱层析进一步分离纯化,获得3个纯化组分(f1、f2、f3)。对f1、f2、f3的纯度进行鉴定,结果表明,f1、f2、f3为均一的多糖组分。采用苯酚-硫酸法、硫酸-间羟联苯法、考马斯亮蓝法、氯化钡-明胶法分别对CP、f1、f2、f3中总糖、糖醛酸、蛋白质、硫酸基含量进行分析。结果表明,CP、f1、f2、f3总糖质量分数分别为83.81%、98.75%、95.58%、84.71%;糖醛酸质量分数分别为1.58%、0.16%、0.96%、2.13%;硫酸基质量分数分别为0.92%、1.22%、2.08%、3.58%;CP、f3蛋白质分别为3.08%、6.34%,f1、f2均未能检出;HPLC分析结果表明,f1、f2、f3的平均相对分子质量分别为68 600、80 600、100 600。采用化学方法对CP、f1、f2、f3进行体外抗氧化活性测定。结果表明,CP、f1、f2、f3具有不同的还原力、脂质过氧化抑制活性、金属离子螯合能力。
|
The polysaccharide from Chinese Venus was firstly separated through acolumn of DEAE-cellulose(Whatman DE 52). As a result,three independent elution peaks(F1,F2and F3)were obtained. The three fractions were loaded onto a column of Sephadex G-100,affording f1,f2and f3,respectively. Purity of f1,f2and f3was further confirmed by using HPLC,and results showed that f1,f2and f3were homogeneous polysaccharides respectively. The chemical characterizations of polysaccharides were investigated by various methods. As to CP,f1,f2and f3,polysaccharides content,measured by employing the method of sulfuric acid-phenol coloration,was 83.81%,98.75%,95.58% and 84.71%,respectively. Uronic acid content,determined by using the method of sulfate-3-phenylphenylol assay,was 1.58%,0.16%,0.96% and 2.13%,respectively. Sulfuric radical content,determined by employing the method of gelatin-barium chloride assay,was 0.92%,1.22%,2.08% and 3.58%,respectively. Protein content,measured by applying the method of coomassie brilliant blue coloration,was 3.08%,not detected,not detected and 6.34% respectively. The relative molecular weight of f1,f2and f3,determined by HPLC,was 68 600,80 600 and 100 600,respectively. The CP,f1,f2and f3had different reducing power,lipid peroxidation inhibition effect and ferrous ion chelating activity.
|
