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白术多糖的分子量及其单糖组成分析
王少杰,杨鸿
0
(佛山科学技术学院生命科学与工程学院)
摘要:
为了建立一种对白术多糖的分子量及其单糖组成测定的方法。采用水提醇沉法提取多糖,提取条件为提取温度90 ℃、料液比1∶16、提取时间1.5 h,重复提取3次;AB-8大孔树脂纯化。采用高效凝胶色谱法测定白术多糖的分子量,流动相:超纯水,流速:0.7 mL/min,柱温:35 ℃,示差折光检测器检测;柱前衍生化HPLC法检测白术多糖的单糖组成,多糖样品用三氟乙酸水解,1-苯基-3-甲基-5-吡唑啉酮柱前衍生化,经C18色谱柱分离,以磷酸盐缓冲溶液-乙腈(85∶15)洗脱,柱温40 ℃,流速1.0 mL/min,进样量20 μL检测波长254 nm。结果显示白术多糖中95%为Mw≈3 500的小分子多糖;经柱前衍生化HPLC法检测,白术多糖由D-甘露糖、D-葡萄糖、D-半乳糖、阿拉伯糖组成,其物质的量比为1∶239∶18.6∶19.3。结果表明试验建立的白术多糖单糖组成分析方法稳定、可靠,可作为白术多糖质量控制方法的参考。
关键词:  白术多糖  分子量  特征性单糖  质量控制
DOI:
投稿时间:2018-11-26修订日期:2019-04-28
基金项目:广东省教育厅预防兽医学重点实验室项目(2014KTSPT037);广东省省级科技计划项目(项目编号2016A020210140)
Analysis of Molecular Weight and Monosaccharide Composition of Atractylodes Macrocephala Polysaccharides
(College of life science and engineering, Foshan University)
Abstract:
A method for the determination of molecular weight and monosaccharide composition of polysaccharide from atractylodes atractyloides was established. Polysaccharides were extracted by water extraction and alcohol precipitation, the extraction conditions were as follows: extraction temperature 90 ℃、material-liquid ratio 1:16、extraction time 1.5 h、repeated extraction 3 times; Purification of Polysaccharides by AB-8 Macroporous Resin. Determination of molecular weight of Atractylodes macrocephala polysaccharides by high performance gel chromatography, Mobile phase: ultrapure water, Flow velocity: 0.7 mL/min, Column temperature: 35 ℃,detection using differential refractive index detector; Using pre-column derivatization HPLC to measure the monosaccharide composition of polysaccharides. The polysaccharide samples were hydrolyzed in trifluoroacetic acid solution, then precolumn derivatizated with 1-phenyl-3-methyl-5-pyrazolone, The chromatographic separation was carried out on Wondasil C18 (250mm×4.6mm, 5μm) using phosphoric acid aqueous solution-acetonitrile ( pH=6.9 ) and acetonitrile ( 85 : 15 ) as mobile phases, at a flow rate of 1 mL/min, 40 ℃, with 254 nm detected wavelength. The results showed:Through HPLC-GPC, 95% of Polysaccharide from Atractylodes macrocephala was Mw≈3 500 small molecule polysaccharide; By pre-column derivatization HPLC, Atractylodes macrocephala polysaccharides are composed of 4 kinds of monosaccharides, D- mannose, D- glucose, D- galactose and Arabia sugar. The monosaccharide composition analysis method of Atractylodes macrocephala polysaccharides established in this experiment is stable and reliable, and can be used as a reference for the quality control of Atractylodes macrocephala polysaccharides.
Key words:  atractylodes macrocephala polysaccharide  molecular weight  characteristic monosaccharide  quality control

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