那屈肝素钙注射液重均相对分子质量与相对分子质量分布探析

Analysis of molecular weight and molecular weight distribution of Nadroparin Calcium injection

目的:考察那屈肝素钙注射液重均相对分子质量与相对分子质量分布情况及存在问题,并对现行标准检验方法进行改进与统一以便更有效评价产品质量。方法:按照现行标准对市售的4家企业47批样品的相对分子质量与相对分子质量分布项目进行检验;改进并统一检验方法:采用TSKgel G2000SWXL(300 mm×7.8 mm)色谱柱分离,以0.1 mol·mL^(-1)醋酸铵溶液为流动相,流速0.5 mL·min-1,柱温30℃,样品浓度1 500 IU·mL^(-1),示差折光检测器对47批样品进行测定;比较分析法定方法和拟定方法对相对分子质量与相对分子质量分布测定结果的影响及不同企业样品相对分子质量与相对分子质量分布的差异;比较分析加速时间对样品相对分子质量与相对分子质量分布的影响。结果:4家企业47批那屈肝素钙注射液的相对分子质量与相对分子质量分布按法定标准检验均符合规定。拟定方法经方法学验证,样品主峰与溶剂峰分离度良好,精密度RSD为0.1%;重均相对分子质量与相对分子质量分布结果的重复性RSD依次为0.1%、1.5%、0.1%、0.1%;日间精密度RSD依次为0.2%、1.9%、0.4%、0.4%。47批样品的相对分子质量与相对分子质量分布按拟定方法检验均符合规定,法定方法与拟定方法测定结果略有差异。B企业、C企业样品相对分子质量小于2 000的级分所占比例较D企业高。A、B、C 3家企业样品重均相对分子质量与加速时间呈负相关性。结论:A、B、C 3家企业样品的相对分子质量与相对分子质量分布及加速试验稳定性较D企业样品有一定差距。1家企业的现行质量标准较低,亟待提高。拟定方法降低流动相盐浓度,减少对色谱柱的损害,操作简便,检验成本低,重复性良好,方法耐用性符合要求,结果准确可靠,可用于那屈肝素钙注射液相对分子质量与相对分子质量分布的测定。

OBJECTIVE To evaluate the quality status and problems of Nadroparin Calcium injection in molecular weight and molecular weight distribution and improve the current methods of evaluating product quality.METHODS The molecular weight and molecular weight distribution of 47 batches of samples from 4 manufacturers were tested by the current standard. Unified and modified method: TSKgel G2000 SWXL(300 mm×7.8 mm)columns were employed for separating the samples. 0.1 mol·mL^(-1) ammonium acetate was utilized as the mobile phase, the flow speed was 0.5 mL·min-1, the column temperature was set at 30 ℃ and the sample concentration was 1 500 IU·mL^(-1). The refractive index detector was used for measuring the results. Two methods were compared with regards to the influences of results and differences of molecular weight and molecular weight distribution results. The effects of acceleration time on molecular weight and molecular weight distribution of samples were compared.RESULTS All samples complied with the general rules of statutory standards. An excellent separation existed between major sample and solvent peaks with a RSD of 0.1%. The results of repeatability for molecular weight and molecular weight distribution were 0.1%, 1.5%, 0.1% and 0.1% and the results of inter-day precision 0.2%, 1.9%, 0.4% and 0.4% respectively. The results for molecular weight and molecular weight distribution of 47 batches of samples from 4 manufacturers complied with the method of formulate standards. The results obtained by two methods were slightly different. The proportion of fractions with molecular weight <2 000 in the samples of B manufacturer and C manufacturer was higher than that of D manufacturer. A negative correlation existed between acceleration time and molecular weight of three manufacturers.CONCLUSION The quality standard of one manufacturer was low and it should be improved. The method of drafting offers the advantages of a low cost, a simple handling, an excellent repeatability and accurate and reliable results. A lower concentration of salt in mobile phase may reduce the damage to chromatographic column. Some gaps in molecular weight and molecular weight distribution and accelerated stability existed among four enterprise samples. The above method is suitable for determining molecular weight and distribution of Nadroparin Calcium injection.