低分子量肝素核磁共振氢谱鉴别法的改进

Improvement of proton nuclear magnetic resonance identification of low molecular weight heparins

目的:改进《美国药典》第42版(USP 42)中低分子肝素(达肝素钠)核磁共振氢谱(~1H NMR)鉴别方法,为低分子量肝素质量标准的制定提供参考。方法:选择3种低分子量肝素(达肝素钠、依诺肝素钠和那屈肝素钙)作为研究对象,比较不同冻干时间和次数与不进行冻干处理的样品的~1H NMR谱,以及有水峰压制实验与无水峰压制实验的样品的~1H NMR谱。结果:利用~1H NMR法对达肝素钠、依诺肝素钠和那屈肝素钙3种低分子量肝素进行鉴别时,是否进行冻干处理和是否选择水峰压制实验的样品的整体谱图和特征峰信号无明显差异,不影响鉴别结果。结论:可以对现有药典低分子量肝素的~1H NMR鉴别方法进行改进,即不进行冻干处理和水峰压制实验。改进后的方法简单易操作,大大提高了效率,为3种低分子肝素(达肝素钠、依诺肝素钠和那屈肝素钙)制订~1H NMR鉴别方法提供参考。

Objective:To improve the proton nuclear magnetic resonance(~1 H NMR)identification method of low molecular weight heparins(LMWHs)(dalteparin sodium)in the United States Pharmacopoeia 42 nd edition(USP 42)to provide reference for the establishment of LMWHs quality standard.Methods:Three types of low molecular heparins(dalteparin sodium,enoxaparin sodium and nadroparin calcium)were selected as the research objects,and the^1 H NMR spectra of samples without freeze-dried operation were compared with those with different freezedried time and numbers,and the^1 H NMR spectra of samples with or without water-suppression experiments were also compared.Results:Freeze-dried operation and water-suppression experiment had no influence on the overall spectrum and characteristic peak of three kinds of LWMHs.With or without the two operations,neither would not affect the judgment of the identification results.Conclusion:The original^1 H NMR identification method of LMWHs in the pharmacopoeia can be improved,that is,no freeze-dried operations and water-suppression experiment are compared.Results:Freeze-dried operation and water-suppression experiment had no influence on the overall spectrum and characteristic peak of three kinds of LWMHs.With or without the two operations,neither would not affect the judgment of the identification results.Conclusion:The original^1 H NMR identification method of LMWHs in the pharmacopoeia can be improved,that is,no freeze-dried operations and water-suppression experiment are carried out.The method after improvement is simple to operate,which greatly improves the efficiency and provides a reference for the establishment^1H NMR method of three types of LMWHs(dalteparin sodium,enoxaparin sodium and nadroparin calcium).