色谱

色谱 ›› 2012, Vol. 30 ›› Issue (02): 173-183.DOI: 10.3724/SP.J.1123.2011.10005

• 研究论文 • 上一篇    下一篇

低分子量硫酸化多糖的体积排阻色谱法分离及其组成定量分析

吴艳芳1, 李晓鸽2, 车铁军3, 朱智甲1, 康经武2*   

  1. 1. 东华大学化学化工与生物工程学院, 上海 201620; 2. 中国科学院上海有机化学研究所, 上海 200032; 3. 宝鸡石油机械有限责任公司, 陕西 宝鸡 721002
  • 收稿日期:2011-10-10 修回日期:2011-12-08 出版日期:2012-02-28 发布日期:2012-03-22
  • 通讯作者: 康经武,研究员,博士生导师,研究方向为色谱分离. Tel: (021)54925385,
  • 基金资助:

    国家自然科学基金项目(Nos. 21175146, 20975109, 9713021)和国家科技重大专项项目(2011ZX09202-101-10)

Separation and quantitative analysis of the composition of low molecular weight sulfated polysaccharides by size exclusion chromatography

WU Yanfang1, LI Xiaoge2, CHE Tiejun3, ZHU Zhijia1, KANG Jingwu2*   

  1. 1. College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620, China; 2. Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, China; 3. Baoji Oilfield Machinery Co., Ltd., Baoji 721002, China
  • Received:2011-10-10 Revised:2011-12-08 Online:2012-02-28 Published:2012-03-22

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442

被引次数

3

摘要： 建立了用于分离并定量测定低分子量硫酸化多糖中不同糖链数的各个组分分布比例的体积排阻色谱方法。系统考察了流动相的组成、离子强度和pH值、流速、柱温等因素对分离的影响。最佳分离条件: 两支TSK-GEL G2000 SWxl色谱柱(300 mm×7.8 mm)串联,流动相100 mmol/L Na2HPO4-NaH2PO4 (pH 7.0),流速0.5 mL/min,柱温35 ℃,进样量5 μL,样品质量浓度10 g/L。在最佳的分离条件下,可以将低分子量硫酸化多糖样品中不同糖链数的各个组分分离并对各个组分的分布进行了定量分析。用该方法对美国药典标准品(USP)、商品和实验室制备的低分子量硫酸化多糖糖链数分布进行了定量化比较,证明该方法可用于低分子量硫酸化多糖类药物的组成成分的质量控制。

关键词: 低分子量硫酸化多糖, 体积排阻色谱, 质量控制, 组成分布

Abstract: As the composition of low molecular weight sulfated polysaccharides is very complex, the quantitative analysis of their compositions is very important for understanding their bioactivity and the purpose of the quality control. A method was developed for the separation and quantitative analysis of the composition of low molecular weight sulfated polysaccharides. The effects of the parameters, such as the mobile phase constitution including ionic strength and pH, column length and temperature, flow rate of the mobile phase on the separation were systematically investigated. The obtained optimal conditions were as follows: two TSK-GEL G2000 SWxl columns (300 mm×7.8 mm) coupled together; mobile phase, 100 mmol/L Na2HPO4-NaH2PO4 (pH 7.0); flow rate, 0.5 mL/min; column temperature, 35 ℃; injection volume, 5 μL; sample concentration, 10 g/L. The method was validated in terms of its reproducibility and robustness. Under the optimized chromatographic separation conditions, each composition of the low molecular weight sulfated polysaccharide can be clearly separated, and their distribution ratios were quantitatively analyzed. The composition profilings of the samples from United States pharmacopoeia (USP), the two commercial available samples and two home-made samples were quantitatively compared. The method can be used for the quality control of the drugs based on low molecular weight sulfated polysaccharides.

Key words: composition profilings, low molecular weight sulfated polysaccharides, quality control, size exclusion chromatography (SEC)