肝素钠相对分子质量对照品国际协作标定Ⅱ

The Second International Collaborative Study on Molecular Weight of Heparin Sodium Reference Standard

目的将第一阶段协作标定的对照品用于凝胶色谱法测定肝素钠相对分子质量,并对凝胶色谱法进行评估。方法凝胶色谱法(GPC),色谱柱为TSK G4000SWXL(7.8 mm×30 cm)和TSK G3000SWXL(7.8 mm×30 cm)串联,流动相为0.1 mol·L-1的乙酸铵(含0.02%叠氮化钠),流速为0.6 m L·min-1±0.1%,色谱柱平衡2 h后进样,柱温为30℃,进样体积20μL,检测器为示差折光检测器。结果本实验室测定结果均通过系统适用性实验,在报告值中间。结论所标定的对照品和考察的凝胶色谱法已收载于USP37中,本次标定的思路和方法为其他多糖相对分子质量对照品的标定提供了很好的参考。

OBJECTIVE To apply the heparin sodium molecular weight reference standard calibrated in the first study period to CPC method and assesse the CPC method. METHODS CPC methods were used. The columns were G4000SW× L （ 7.8 mm × 30 cm）-TSK G3000SWxL（7.8 mm× 30 cm） in series maintained at 30 ℃. The mobile phase was 0. 1 mmol· L^-1 ammonium acetate so- lution containing 0. 02% NaN3 eluted at a flow rate of 0. 6 mL · min^-1. The samples were injected after a column equilibration period of 2 h and the injection volume was 20 μL. The analysis was performed by RI detector. RESULTS The analysis result passed the system suitability test, with measured values in the middle of the report. The precision and accuracy were good. The reference standard and GPC method calibrated in this study have already been recorded in USP37. CONCLUSION The methods and ideas we used are suitable for the calibration of other polysaecharides.