玻璃酸钠及制剂分子质量与分子质量分布测定方法研究

Determination of molecular weight and molecular weight distribution of sodium hyaluronate and its preparations

目的:分析玻璃酸钠(SH)及制剂分子质量和分子质量分布测定不同模式的特点,重点研究折光指数增量(dn/dc值)、对照品等对测定结果的影响。方法:采用凝胶色谱法(SEC),色谱柱为ShodexSB806HQ(300 mm×8 mm),流动相为0.2 mol·L-1的氯化钠溶液(含0.02%叠氮化钠),流速为0.5 mL·min-1,柱温为35℃,进样量100μL,检测采用2种方法,单一示差折光检测器及采用示差折光检测器和激光光散射仪(MALLS)联用。结果:玻璃酸钠及制剂相对分子质量和绝对分子质量测定结果存在约30%的误差,dn/dc值、对照品等对测定结果产生一定影响。结论:凝胶色谱示差折光检测法和凝胶色谱激光光散射联用法,2种方法均可用于玻璃酸钠及制剂分子质量控制。

Objective： To analyze the different determination modes of molecular weight and molecular weight distribution of sodium hyaluronate（ SH） and its preparations,especially the influence of dn /dc value and standards on the determination results. Methods： Using SEC method,samples were analyzed on a Shodex SB806 HQ column（ 300 mm ×8 mm）;the mobile phase was 0. 2 mol·L- 1sodium chloride（ containing 0. 02% sodium azide） at a flow rate of 0. 5 mL·min- 1;the injection was 100 μL; the column temperature was 35 ℃,and the signals were detected by RI detector（ used by SEC method） or RI coupled with MALLS detectors（ used by SEC- MALLS method）. Results： The results showed that there were about 30% differences between the molecular weight and molecular weight distribution results determined by SEC-RI and SEC- MALLS methods,and the dn /dc value and standards had certain influences on the determination results.Conclusion： SEC- RI and SEC- MALLS methods could both be used for quality control of SH and its preparations.