Analysis of Monosaccharide Compositions in Suanzaoren Decoction Polysaccharides by Pre-column Derivatization High Performance Liquid Chromatography

LIU Yu; LOU Yan; ZHANG Yao-wen; WANG Tie-jie; BI Kai-shun; LI Qing

doi:10.11653/syfj2013170121

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Analysis of Monosaccharide Compositions in Suanzaoren Decoction Polysaccharides by Pre-column Derivatization High Performance Liquid Chromatography

更新时间：2024-01-04

- Analysis of Monosaccharide Compositions in Suanzaoren Decoction Polysaccharides by Pre-column Derivatization High Performance Liquid Chromatography
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 19, Issue 17, Pages: 121-126(2013)
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- DOI：10.11653/syfj2013170121
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- Published：2013
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LIU Yu, LOU Yan, ZHANG Yao-wen, et al. Analysis of Monosaccharide Compositions in Suanzaoren Decoction Polysaccharides by Pre-column Derivatization High Performance Liquid Chromatography[J]. Chinese journal of experimental traditional medical formulae, 2013, 19(17): 121-126.
DOI：

LIU Yu, LOU Yan, ZHANG Yao-wen, et al. Analysis of Monosaccharide Compositions in Suanzaoren Decoction Polysaccharides by Pre-column Derivatization High Performance Liquid Chromatography[J]. Chinese journal of experimental traditional medical formulae, 2013, 19(17): 121-126. DOI： 10.11653/syfj2013170121.

摘要

目的: 建立1-苯基-3-甲基-5-吡唑琳酮(PMP)柱前衍生高效液相色谱法分离检测4种单糖和2种糖醛酸的方法。方法: 采用水提醇沉法提取酸枣仁汤中多糖

中空纤维超滤膜法纯化

然后用硫酸水解

1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生后

利用反相高效液相色谱法

分析单糖的PMP衍生物。以单糖PMP衍生物的峰面积大小为指标

分别对衍生化反应中反应时间、PMP与NaOH的用量、中和温度和盐酸用量进行考察

从而得到较佳的样品衍生化制备条件。并将该方法应用于酸枣仁汤中总多糖 (SDP)、相对分子量<10 kDa多糖(SDP-1)、相对分子量为10~50 kDa多糖(SDP-2)、相对分子量>50 kDa多糖(SDP-3)的单糖组成分析和含量测定。结果: SDP和SDP-1均由Man-GlcUA-GalUA-Glc-Gal-Ara组成

摩尔比分别为1:0.8:0.7:3.4:3.3:2.1和1:0.5:0.3:0.6:0.6:0.4。SDP-2 由Man-Glc-Gal-Ara组成

摩尔比为1:2.0:0.2:0.3;SDP-3由Man-Glc组成

摩尔比为1:5.4。结论: 方法准确、灵敏度高

适用于酸枣仁汤中多糖的单糖组成测定。

Abstract

Objective: A method of 1-phenyl-3-methyl-5-pyrazolone (PMP) pre-column derivatization high performance liquid chromatographic was established for analysis of four kinds of monosaccharide and two kinds of galacturonic acid and glucouronic acid. Method: The polysaccharides were extracted by hot distilled water

precipitated by alcohol. The product was purified by ultrafiltration and formed monosaccharides by sulfuric acid hydrolysis. The hydrolysate reacted with PMP to produce the PMP derivatives that were analyzed by HPLC. The optimum conditions of derivatization were determined by studying the quantity of PMP and sodium hydroxide

reaction time

reaction temperature and hydrochloric acid consumption during neutralization. Under the optimum mobile phase which was acetonitrile-phosphate buffer (pH 6.8)

PMP derivatives can be separated successfully. This method was applied to monosaccharide compositions analysis of SDP

SDP-1

SDP-2

SDP-3. Result: The results showed that three polysaccharides (SDP-1

SDP-2

SDP-3) were obtained by ultrafiltration

molecular weights were less than 10

10-50 kDa and more than 50 kDa respectively. Both SDP and SDP-1 were composed of Man-GlcUA-GalUA-Glc-Gal-Ara in molar ratio of 1: 0.8: 0.7: 3.4: 3.3: 2.1 and 1: 0.5: 0.3: 0.6: 0.6: 0.4;SDP-2 was composed of Man-Glc-Gal-Ara in molar ratio of 1: 2.0: 0.2: 0.3;SDP-3 was composed of Man-Glc in molar ratio of 1: 5.4. Conclusion: The method is accurate and sensitive

thus suitable for the analysis of monosaccharide composition of Suanzaoren Decoction polysaccharide.

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Related Institution

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