摘要
目的寻找佐剂茯苓多糖PCP-Ⅰ部分酸水解的最佳条件,为制备其主要寡糖片段和结构解析提供实验依据。方法采用薄层层析法和高效凝胶渗透法,首先考察不同三氟乙酸(TFA)浓度(0.025~0.5 mol/L)、不同温度(40、60、80、100和120℃)、不同水解时间(1、2、4、6和8 h)对酸水解产物的影响;其次采用L9(34)正交设计进行三因素和三水平的试验和综合分析,最终确定PCP-Ⅰ最佳酸水解的条件参数。结果单因素考察结果显示:(1)在80℃和水解2 h条件下,0.05~0.1 mol/L TFA适合水解;(2)在0.05 mol/L TFA和水解2 h条件下,80~100℃适合水解;(3)在0.1 mol/LTFA和100℃条件下,1~4 h适合水解。正交试验结果表明,100℃条件下,0.05 mol/L TFA水解4 h或0.1 mol/L TFA水解2 h,产物主要含有1745(Mw)或1838为寡糖和少量单糖。结论这两种水解条件操作简单、寡糖含量高,适合PCP-Ⅰ酸水解及制备主要寡糖片段。
Objective To select optimal conditions for partial acid hydrolysis of polysaccharide adjuvant PCP-Ⅰfrom Poria cocos in order to prepare a main oligosaccaride fragment from the polysaccharide.Methods Firstly,the acid hydrolysates were splitted by different concentrations of TFA(0.025-0.5 mol/L),temperatures(40,60,80,100 and 120℃)and reaction time(1,2,4,6 and 8 h).Analysis was conducted using thin layer chromatography(TLC)analysis and high performance liquid chromatography(HPGPC).Secondly,a L9(34)orthogonal design of three factors and three levels was used to study acid hydrolysis of PCP-Ⅰbefore an optimal hydrolysis condition was determined.Results As was shown by the results of single factor experiments,(1)under 80℃and after two hours of hydrolysis,0.05-0.1 mol/L of TFA was suitable;(2)under 0.05 mol/L TFA and after two hours of hydrolysis,80-100℃was suitable;(3)under 100℃and 0.1 mol/L TFA for hydrolysis,1-4 h was desirable.The results of orthogonal design showed that these two conditions were ideal for acid hydrolysis of PCP-Ⅰand that two oligosaccharides were produced respectively(Mw=1745 or 1838).The two optimum conditions for acid hydrolysis of PCP-Ⅰwere 0.05 mol/L TFA and 4 h,or 0.1 mol/L TFA and 4 h both under100℃.Conclusion The two conditions are easy to operate and optimal for acid hydrolysis of PCP-Ⅰ,which can be adopted to prepare a main oligosaccharide fragment.
作者
刘桂馨
麻浩
姚蕙
韩笑
单俊杰
LIU Gui⁃xin;MA Hao;YAO Hui;HAN Xiao;SHAN Jun⁃jie(Nanjing University of Chinese Medicine,Nanjing 210046,China;Institute of Pharmacology and Toxicology,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China)
出处
《军事医学》
CAS
北大核心
2020年第12期930-936,共7页
Military Medical Sciences
关键词
茯苓多糖
部分酸水解
薄层色谱
高效凝胶渗透色谱
polysaccharide of Poria cocos
partial acid hydrolysis
thin layer chromatography
high performance liquid chromatography
