Chem. J. Chinese Universities ›› 2010, Vol. 31 ›› Issue (3): 479.

• Articles • Previous Articles     Next Articles

Analysis of Oversulfated Chondroitin Sulfate in Contaminated Heparin by Precolumn Derivatization High Performance Liquid Chromatography

ZHAO Xia*, LI Guang-Sheng, YU Guang-Li, WANG Jin-Xia, WANG Hao, SUN Shu-Meng, ZHENG Chen   

  1. Key Laboratory of Marine Drugs, Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China
  • Received:2009-10-19 Online:2010-03-10 Published:2010-03-10
  • Contact: ZHAO Xia. E-mail: zhaoxia@ouc.edu.cn
  • Supported by:

    国家自然科学基金(批准号: 30870506)和国际科技合作项目(批准号: 2007DFA30980)资助.

Abstract:

Based on the monosaccharide composition difference between heparin and OSCS, a simple and quantitative high performance liquid chromatography method has been established to determine OSCS present in contaminated heparin in this paper. After degradation of contaminated heparin by 3 mol/L trifluoroacetic acid(TFA) at 110 ℃ for 4 h, the monosaccharides were derivatized with 1-phenyl-3-methyl-5-pyrazolone(PMP), and separated on a C18 reversed phase column with a mobile phase of 0.1 mol/L phosphate buffer-acetonitrile(82∶18, volume ratio) at a column temperature of 25 ℃. The flow rate was 1.0 mL/min and the detection wavelength was 245 nm. There is a good separation between the peaks of monosaccharide-PMP derivatives of heparin and OSCS under the chromatographic conditions, and the two lots of contaminated heparin were found to contain 19.6% and 28.3% OSCS, respectively. This method is easy to apply and suitable for the determination of OSCS contaminant in heparin with high accuracy, reproducibility and sensitivity.

Key words: Heparin; Oversulfated chondroitin sulfate; High performance liquid chromatography; 1-Phenyl-3-methyl-5-pyrazolone

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