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

Based on the monosaccharide composition difference between heparin and Oversulfated chondroitin sulfate(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.