大孔吸附树脂分离纯化戴氏虫草多糖工艺研究  

Studies on the separation and purification of Metarhiziun taii polysaccharide by macroporous adsorption resin

作　　者：肖代敏[1] 吕纯莉[1] 曹喻[1] 肖建辉[1] 张志敏[2] 

机构地区：[1]遵义医学院附属医院医学检验科,贵州遵义563099 [2]遵义医学院遗传学教研室,贵州遵义563099

出　　处：《遵义医学院学报》2015年第5期536-540,共5页Journal of Zunyi Medical University

基　　金：贵州省科技厅基金资助项目(NO:黔科合J字[2009]2148);贵州省科技厅联合基金资助项目(NO:黔科合J字LKZ[2011]35)

摘　　要：目的以多糖含量为考核指标,筛选分离纯化戴氏虫草（Metarhiziun taii,M.taii）多糖最佳吸附树脂,探讨其最佳吸附工艺,建立M.taii多糖分离纯化方法。方法从8种吸附树脂中筛选最佳吸附大孔树脂;通过静态吸附曲线和解吸附曲线确定树脂对多糖吸附与解吸附饱和情况,不同洗脱剂洗脱后采用L9（3）4正交实验对树脂吸附主要相关因素（多糖浓度、pH、温度及振荡频率）进行考察,确定大孔树脂吸附及解吸附最优条件;并进行动态吸附实验考察树脂最大吸附能力;采用不同洗脱剂进行连续梯度洗脱以观察动态洗脱情况。结果 8种不同树脂中,D101大孔树脂最适合M.taii多糖的分离纯化,约5 h静态吸附与解吸附达到饱和,最佳洗脱剂为60%乙醇溶液,最优条件为pH 6.0、20℃、低浓度多糖液（2.5-3.0mg/mL）、每15分钟混匀1次。经验证,多糖吸附率、解吸附率及脱色率分别为（71.85±1.12）%、（74.37±2.94）%和（64.84±1.89）%;动态吸附结果表明,每1 g树脂吸附7.79 mg多糖达到饱和;M.taii多糖液经D101大孔树脂柱层析纯化,用纯水和15%乙醇溶液就能将大部分多糖洗脱。结论 D101大孔树脂是一种较理想的分离纯化虫草多糖的树脂,为虫草多糖的分离纯化提供了实验依据。Objective To screen the best adsorption resin for separation and purification of M. taii polysaccharide by testing polysaecharide content as evaluation index, and to study the best adsorption technology, the methods of separation and purification of M. tail polysaccharide were established. Methods selecting macroporous resin with best adsorption from 8 kinds of adsorption resin ; static adsorption curve and desorption curve were used to determine saturated state of polysaccharide＇ s adsorption and desorption in resin, L9 （ 3 ）^ 4 orthogonal experiment was used to investigate mainly related factors of resin adsorption （concentration of polysaceharide, pH, temperature and oscillation frequency） after elution by different eluents, to obtain optimal conditions of adsorption and desorption about rnacroporous resin; At the same time dynamic adsorption experiments used to find out the maxi- mum adsorption capacity of resin ; Using different eluents with continuous gradient elution to observe the dynamic elution conditions. Results D101 macroporous resin from 8 kinds of resin was the most fitful for separation and purification of M. taii polysaceharide, the static adsorption and desorption were saturated about 5 h later, the best elution was 60% ethanol solution, the optimal conditions were pH 6.0, room temperature, low concentration of polysaceharide （2.5 -3.0 mg/mL） and mixing in every 15rain. It were conformed that rate of polysaccharide adsorption, resolution and deeolorization were （71.85 ± 1.12）%, （74.37 ± 2.94）% and （64.84 ±1.89） % respectively ; dynamic adsorption results show that it could reach saturation when 1 g resin adsorpted 7.79 mg polysaecharide; most M. taii polysaceharide could be eluted by DI01 maeroporous resin column chromatography purification then with pure water and 15% ethanol solution. Conclusion D101 maeroporous resin is a kind of excellent resin for separation and purification of Cordyeeps polysaccharide, the study provides the experimental basis for the separatio

关 键 词：大孔树脂 虫草多糖 吸附 解吸附 分离纯化 

分 类 号：R961[医药卫生—药理学]