三斑海马多糖的单糖组成测定影响因素考察  

The influence of hydrolysis conditions on monosaccharide composition determination of polysaccharide from Hippocampus trimaculatus.

作　　者：王清池[1] 赵峡[1] 邓世明[2] 

机构地区：[1]中国海洋大学海洋药物教育部重点实验室,山东省糖科学与糖工程重点实验室,医药学院,山东青岛266003 [2]海南大学海洋学院,海南海口570100

出　　处：《中国海洋药物》2016年第2期30-34,共5页Chinese Journal of Marine Drugs

基　　金：“十二五”国家科技支撑计划项目(2013BAB01B02,2013BAB01B04);国家基金委山东联合基金项目(U1406402)资助

摘　　要：目的研究不同因素对三斑海马多糖(HTP)降解的影响,探求适合其单糖组成测定的降解条件。方法通过设计正交实验,采用三氟乙酸(TFA)对HTP进行降解,结合1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生高效液相色谱进行单糖组成分析,比较各实验的降解情况。结果 HTP中主要含有Man、GlcN、Gal和少量的GalN、Glc、GlcA、Ara、Fuc共8种单糖。在考察条件范围内,温度和时间是影响单糖组成分析的主要因素,酸浓度是次要因素。HTP中各单糖的最适降解条件是:氨基糖GlcN和GalN为120℃,6h,4mol/L TFA;中性糖Man、Glc和Gal为110℃,4h,3mol/L TFA;糖醛酸GlcA及Ara、Fuc为100℃,2h,3mol/L TFA。通过对10种单糖标准品的降解也发现氨基糖(GlcN和GalN)较稳定,糖醛酸(GlcA和GalA)稳定性差。结论HTP中不同结构类型的单糖对降解条件的敏感度不同:糖醛酸易降解释放,但也易被破坏;氨基糖较难降解,但降解后较为稳定;中性糖的降解难易和稳定性介于两者之间。采用同一条件降解不能准确地测定HTP的单糖组成,需要针对不同结构类型单糖的性质采用不同的条件进行降解。Objective To find the appropriate hydrolysis condition on monosaccharide composition determination of polysaccharide from Hippocampus trimaculatus（HTP）by investigating the influence of the hydrolysis conditions.Methods Orthogonal experiment was designed to compare the degree of hydrolysis of HTP by trifluoroacetic acid（TFA）degradation and high performance liquid chromatography（HPLC）analysis with pre-column derivatization.Results HTP contained eight monosaccharides,which mainly were Man,GlcN,Gal,and a bit of GalN,Glc,GlcA,Ara,Fuc.The hydrolysis temperature and time were the major factors on the polysaccharide hydrolysis,while concentration of TFA was the minor factor on it.The appropriate hydrolysis conditions of different monosaccharides in HTP were as follows：Aminosaccharides（GlcN and GalN）were 120 ℃,6h,4mol/L TFA.Neutral sugars（Glc,Gal and Man）were 110 ℃,4h,3mol/L TFA.Uronic acid（GlcA）,Fuc and Ara were 100 ℃,2h,3mol/L TFA.Aminosaccharides had good stability,while the uronic acids showed poor stability by hydrolyzing ten monosaccharide standards.Conclusion Different monosaccharides of HTP had different sensitivity to hydrolysis conditions.Uronic acid was much easily hydrolyzed and destroyed,while aminosaccharides were hard to hydrolyze but stable when they were hydrolyzed.The stability and hydrolysis degree of neutral sugars were in between uronic acid and aminosaccharides.Thus,different hydrolysis conditions should be used for the monosaccharide composition determination of HTP based on the different characteristics of each monosaccharide.

关 键 词：三斑海马 多糖 单糖组成 柱前衍生 降解条件 

分 类 号：R931[医药卫生—生药学]