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期刊名称:药物分析杂志
主管单位:中国科学技术协会
主办单位:中国药学会
承办:中国食品药品检定研究院
主编:金少鸿
地址:北京天坛西里2号
邮政编码:100050
电话:010-67012819,67058427
电子邮箱:ywfx@nifdc.org.cn
国际标准刊号:ISSN 0254-1793
国内统一刊号:CN 11-2224/R
邮发代号:2-237
 

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糖谱法结合多元色谱分析比较铁皮石斛功能性多糖结构特征

Characterization and comparison of specific polysaccharides in Dendrobium officinale by using saccharide mapping and chromatographic methods

作者(英文):
分类号:R917
出版年·卷·期(页码):2018,38 (1):41-49
DOI: 10.16155/j.0254-1793.2017.01.01
-----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×0.25 μm),进样量2 μL,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。

-----英文摘要:---------------------------------------------------------------------------------------

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 G3000PWXL in series were employed with the temperature of 35.The mobile phase was 9 mg·mL-1 NaCl aqueous solution at a flow rate of 0.5 mL·min-1.GC-MS was performed on an Agilent HP-5MS 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 aniline-diphenylamine.Results:The weight-average relative molecular mass,radius of gyration,and the content of specific polysaccharide in D. officinale from different locations were 0.533×105-4.575×105,19.8-71.7 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:Similarity of specific polysaccharides in D. officinale from different habitats was high,which could be a good marker for the quality control of D. officinale.

-----参考文献:---------------------------------------------------------------------------------------

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