摘要
目的:比较不同产地铁皮石斛功能性多糖结构特征,为其质量控制奠定基础。方法:高效凝胶色谱联用多角度激光散射和示差折光仪法(HPSEC-MALLS-RID法)用于测定多糖的重均相对分子质量、回旋半径及含量,色谱柱为TSK-GEL G5000 PWXL串联TSK Gel G3000PWXL,柱温35℃,流动相为9 mg·mL^(-1)氯化钠水溶液,流速0.5 mL·min^(-1)。气相色谱联用质谱法用于分析单糖组成及糖苷键,毛细管色谱柱为Agilent HP-5MS(30 m×0.25 mm×0.25μm),进样量2μL,用于单糖组成分析的程序升温条件为起始温度165℃,保持7 min,以5℃·min^(-1)升温至185℃,保持5 min,以4℃·min^(-1)升温至200℃,以20℃·min^(-1)升至280℃;用于糖苷键分析的程序升温条件为起始温度120℃,以5℃·min^(-1)升温至200℃,以8℃·min^(-1)升温至250℃,以20℃·min^(-1)升至280℃。基于荧光辅助凝胶电泳、高效薄层色谱-糖谱法分析功能性多糖部分酸水解和酶解产物特征,荧光辅助凝胶电泳采用200 V分离10 min,再调整电压至700 V分离40 min,凝胶在UV 365 nm下成像;TLC展开剂为正丁醇-异丙醇-水-醋酸(7∶5∶2∶1),显色剂为苯胺-二苯胺。结果:不同产地铁皮石斛多糖重均相对分子质量、回旋半径和含量分别为0.533×105~4.575×105、19.8~71.7 nm和6.28%~28.72%;铁皮石斛功能性多糖的单糖组成主要为(乙酰)甘露糖和葡萄糖,摩尔比(3.86~6.99)∶1.00,糖苷键类型主要为1,4-Manp和1,4-Glcp,摩尔比(4.51~7.89)∶1.00;不同铁皮石斛多糖的部分酸水解和酶解产物糖谱特征相似。结论:不同产地铁皮石斛功能性多糖结构具有较高的一致性,功能性多糖是铁皮石斛较好的质量控制指标。
Objective：To characterize and compare specific polysaccharides in Dendrobium officinale from different habitats for its quality control.Methods：HPSEC-MALLS-RID was adopted for the analysis of molecular weights,radius of gyrations and contents of specific polysaccharide.Columns of TSK-GEL G5000 PWXL and TSK Gel G3000 PWXL in series were employed with the temperature of 35 ℃.The mobile phase was 9 mg·mL^-1 Na Cl aqueous solution at a flow rate of 0.5 mL·min^-1.GC-MS was performed on an Agilent HP-5 MS capillary column（30 m×0.25 mm×0.25 μm） to analyze compositional monosaccharides and glycosidic linkages.Injection volume was 2 μL.For compositional monosaccharides analysis,the column temperature was set at 165 ℃ and held for 7 min then programmed at 5 ℃·min^-1 to 185 ℃ and held for 5 min,then at 4 ℃·min^-1 to 200 ℃,and finally,at 20 ℃·min^-1 to 280 ℃.For glycosidic linkages analysis,the column temperature was set at 120 ℃ and then programmed at 5 ℃·min^-1 to 200 ℃,then at 8 ℃·min^-1 to 250 ℃,and finally,at 20 ℃·min^-1 to 280 ℃.Saccharide mapping based on PACE and TLC was employed for fingerprinting of partial acid and enzymatic hydrolysates of specific polysaccharides.For PACE analysis,samples were electrophoresed first at 200 V for 10 min,then at 700 V for 40 min and gels were imaged under UV 365 nm.The TLC plate was developed with 1-butanol-isopropanol-acedicacid-water（7∶5∶2∶1） and the colorized with anilinediphenylamine.Results：The weight-average relative molecular mass,radius of gyration,and the content of specific polysaccharide in D. officinale from different locations were 0.533×10^5-4.575×10^5,19.8-71.7 nm,and 6.28%-28.72%,respectively.The main compositional monosaccharides were（acetylated） mannose and glucose（3.86-6.99）∶1.00 with glycosidic linkages of 1,4-Manp and 1,4-Glcp（4.51-7.89）∶1.00.Saccharide mappings of partial acid and enzymatic hydrolysates from specific polysaccharides of different samples were similar.Conclusion：Si
出处
《药物分析杂志》
CAS
CSCD
北大核心
2018年第1期41-49,共9页
Chinese Journal of Pharmaceutical Analysis
基金
国家自然科学基金(No.81673389和No.81603069)
澳门科学技术发展基金项目(074/2016/A2
034/2017/A1和040/2016/A)
澳门大学研究基金(MYRG2015-00202和MYRG2015-00122)
关键词
铁皮石斛
功能性多糖
单糖组成
糖苷键
含量测定
高效凝胶色谱
多角度激光散射
示差折光法
糖谱法
Dendrobium officinale
specific polysaccharides
compositional monosaccharides
glycosidic linkage
content determination
HPSEC
MALLS
RID
saccharide mapping
