响应面法优化茯苓多糖硫酸酯化合成工艺及其抗凝血活性剖析.docVIP

响应面法优化茯苓多糖硫酸酯化合成工艺及其抗凝血活性剖析 　　摘要：以中药茯苓（Poria cocos）为原料提取茯苓多糖，以氨基磺酸为酯化试剂对茯苓多糖进行硫酸酯化结构改性，制备硫酸酯化茯苓多糖（SP），在单因素试验基础上，应用响应面法试验优化其制备条件。对获得的SP进行纯化、结构表征，通过小白鼠体内抗凝血试验考察SP的抗凝血活性。结果表明，最佳的硫酸酯化工艺条件为反应温度85 ℃、反应时间4.0 h、氨基磺酸∶多糖质量比为3.77∶1，在此条件下，硫酸基团取代度（DS）为1.041，与氯磺酸吡啶法相比取代度相当。精制后SP的红外图谱表明，SP具有多糖及硫酸根基团的特征吸收峰;紫外光谱扫描表明SP无核酸、蛋白质等杂质，纯度较高，且有硫酸根基团的特征吸收，硫酸酯化成功。茯苓多糖经硫酸酯化结构改性后表现出良好的抗凝血活性，其出血时间、全凝血时间、全凝血时间延长率均明显提高;SP对凝血酶的抑制作用与其浓度呈正相互作用，随着SP腹腔注射给药剂量增大，其全凝血时间、延长率均明显提高。 　　关键词：茯苓（Poria cocos）;硫酸酯化;茯苓多糖;响应面法;抗凝血活性 　　中图分类号：S567.3+2 文献标识码：A 文章编号：0439-8114（2015）23-5995-06 　　DOI：10.14088/j.cnki.issn0439-8114.2015.23.051 　　Analysis of The Optimization of Sulfation Pachymaran Synthesis Technology Process by Response Surface Method and Sulfation Pachymaran Anticoagulant Activity 　　CHEN Xiao-hong 　　（College of Life Science，Longyan University，Longyan 364012，Fujian，China） 　　Abstract：The pachyman was extracted from traditional Chinese medicine Poria cocos，with amino sulfo-nic acid as esterification reagent， treated the pachyman with sulfated processing to make sulfation pachymaran （SP） and optimized the conditions of their production processing by response surface method. After purification， structural characterization，sulfation pachymaran（SP） was examined the anticoagulant activity by the anticoagulant effect test in mice. Experimental results showed that the optimal sulfated processing of sulfation pachymaran（SP） were as follows： reaction time 4.0 h，chlorousulfonic acid and pyridine mass ration 3.77∶1，reaction temperature 85 ℃. Under those conditions the degree of substitution was 1.041，the degree of substitution was equal compared with the method of chlorosulfonic acid pyridine in the same conditions. The SP has the characteristics of polysaccharides and sulfate group absorption peak in its ir spectrum after refined， the SP showed high purity and no impurities such as nucleic acids， proteins， and had the characteristics of sulfate group absorption in its UV scanned map. Good anticoa