低分子肝素相对分子质量测定用对照品的研制

Study on the preparation of relative molecular mass calibrator for low molecular mass heparin

目的:制备类似WHO的附带一宽分布标样表的低分子肝素(low molecular weight heparin,LMWH)相对分子质量测定用对照品。方法:采用肝素酶Ι控制降解肝素得到宽分布低分子的肝素片段混合物,并以WHO LMWH相对分子质量对照品为对照,采用高效分子排阻色谱法绘制自制相对分子质量与累积面积百分比的量表。分别利用该自制对照品与WHO对照品检测4种不同工艺制备的LMWH相对分子质量。结果:自制LMWH相对分子质量对照品与WHO相对分子质量对照品的校正曲线基本重合在一起,2种对照品测定的4种LMWH相对分子质量结果十分接近。结论:自制的LMWH相对分子质量测定用对照品可以用于LMWH相对分子质量的测定,为LMWH相对分子质量测定用对照品国产化奠定了基础。

Objective： To establish a relative molecular mass calibrator with a broad standard table for low molecu- lar mass heparin （LMWH）. Methods： LMWH relative molecular mass calibrator with broad distribution was pro- duced through controlled degradation of heparin sodium by heparinase I. With the WHO calibrator as the reference standard, a broad standard table for the prepared calibrator was constructed by high performance size exclusive chro- matography （HPSEC）. To evaluate the feasibility of the latter, molecular mass profiles of four LMWHs obtained with different methods were determined by WHO LMWH calibrator and the prepared calibrator. Results： The calibration curves of the prepared calibrator coincided with the WHO calibrator in general. Relative molecular mass of LMWHs determined by the prepared calibrator agreed well with that of the WHO calibrator. Conclusion： The prepared cali- brator was suitable for routine quality control of molecular mass determination for commercial LMWH products.