问题肝素中多硫酸软骨素杂质的柱前衍生高效液相色谱分析  

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

作　　者：赵峡[1] 李广生[1] 于广利[1] 王金霞[1] 王皓[1] 孙淑萌[1] 郑晨[1] 

机构地区：[1]中国海洋大学医药学院,海洋药物教育部重点实验室,青岛266003

出　　处：《高等学校化学学报》2010年第3期479-483,共5页Chemical Journal of Chinese Universities

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

摘　　要：基于肝素和多硫酸软骨素(OSCS)在单糖组成上的差别,建立了可用于肝素中OSCS检测的柱前衍生高效液相色谱法.采用3 mol/L三氟乙酸,将受污染的问题肝素在110℃下充氮封管水解4 h,在碱性条件下与1-苯基-3-甲基-5-吡唑啉酮进行衍生化反应,再采用C18反相色谱柱,以0.1 mol/L磷酸盐(pH=6.7)缓冲液/乙腈(体积比82∶18)为流动相,在流速1.0 mL/min、柱温25℃及紫外检测波长245 nm的条件下进行液相色谱分析.结果表明,肝素和OSCS的单糖色谱峰具有良好的分离度,测得2批问题肝素中OSCS杂质的质量分数分别为19.6%和28.3%.该方法具有良好的精密度和重现性,易于推广,适合于肝素中OSCS杂质的检测,并可用于硫酸软骨素A和C与硫酸软骨素B的区分和鉴别.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.

关 键 词：肝素 多硫酸软骨素 高效液相色谱 1-苯基-3-甲基-5-吡唑啉酮 

分 类 号：O657[理学—分析化学] O629[理学—化学]