猪凝血酶中右旋糖酐20含量测定的高效液相色谱法建立及验证

目的建立测定猪凝血酶中右旋糖酐20含量的高效液相色谱法,并对方法进行验证和初步应用。方法根据分子排阻色谱法,以亲水硅胶高效体积排阻色谱柱进行分离,采用示差折光检测器测定右旋糖酐20含量。应用该方法对3批猪凝血酶样品中右旋糖酐20含量进行检测。结果该方法专属性强,线性范围为2.314~7.406 mg/ml;低、中、高3个浓度水平的平均回收率分别为99.72%、100.60%、101.83%,总平均回收率为100.72%(n=9),相对标准偏差为1.10%(n=9);中间精密度相对标准偏差为1.43%(n=12)。3批猪凝血酶样品中右旋糖酐20含量分别为18.051、18.731、18.161 mg/ml,均符合企业标准。结论该方法专属性强,准确度、精密度、耐用性良好,适用于猪凝血酶中右旋糖酐20含量测定。

Establishment and validation of high performance liquid chromatography determination of dextran 20 in porcine thrombin

Objective To establish a method for determination of dextran 20 in porcine thrombin by high performance liquid chromatography and perform method validation and application.Methods Based on molecular exclusion chromatography and hydrophilic silica gel column with high efficiency volume exclusion chromatography,the content of dextran 20 in porcine thrombin was determined by refractive index detector.The method was applied to determine dextran 20 content in three porcine thrombin batches.Results The method had strong specificity and the linear range was 2.314~7.406 mg/ml.The average recoveries at low,medium and high levels were 99.72%,100.60%and 101.83%,respectively and the overall average recovery was 100.72%(n=9),with the relative standard deviation(RSD)of1.10%(n=9).The RSD obtained from the intermediate precision was 1.43%(n=12).The dextran 20 contents of three porcine thrombin batches were 18.051,18.731,18.161 mg/ml,complied with the enterprise standards.Conclusion This method,with good specificity,accuracy,precision,and durability,is suitable for determination of dextran 20 in porcine thrombin.