HPLC法测定valrubicin膀胱滴注液含量及有关物质.docVIP

HPLC法测定valrubicin膀胱滴注液含量及有关物质 　【摘要】 目的 建立合适的高效液相色谱法测定valrubicin膀胱滴注液中valrubicin的含量及有关物质。方法 采用岛津Shim-pack VP-ODS色谱柱；流动相为0.015mol/L磷酸溶液(取磷酸1ml，加水稀释至1000ml)∶乙腈(43∶57)；柱温35℃；检测波长254nm；流速1.5ml/min。结果 主药与杂质和或降解产物良好分离，空白油辅料无干扰。以本法检测样品中所含0.05%～10%范围的有关物质，其线性关系良好；在0.01～1.18mg/ml范围内valrubicin浓度与峰面积呈良好的线性关系，适合于含量测定。有关物质检测的LOD为0.1μg/ml，相当于可检出所有浓度低至0.01%的杂质和或降解产物。有关物质检测结果重复性好；含量测定重复性试验测得valrubicin平均含量为96.8%(RSD=0.34%，n=9)。Valrubicin含量测定的平均回收率为100.5%(RSD=0.4%，n=9)。结论 本法色谱性能明显优于USP个论方法，专属性强，更适用于valrubicin膀胱滴注液的含量测定和有关物质检测。 【关键词】 Valrubicin； 膀胱滴注液； 高效液相色谱法； 含量测定； 有关物质 ABSTRACT Objective To establish an alternative high performance liquid chromatography method for the determination of valrubicin and its related substances in valrubicin intravesical solution. Methods TheHPLC method was applied with a Shimadzu Shim-pack VP-ODS (4.6mm×150mm, 5μm) column by anelution system consisted of 0.015 mol/L phosphate acid solution (phosphate acid 1ml added to 1000ml water)∶acetonitrile (43∶57); the flow rate was 1.5 ml/min. The detective wavelength was set at UV 210nm; the column temperature was hold at 35℃. Results The good resolution between peaks of valrubicin and the adjacent impurity and/or degradation product was achieved. The blank excipient consisted of polyoxyl 35 castor oil and ethanol (50∶50) did not interfere in the separation and detection of valrubicin. The linear range for analysis of the related substances of valrubicin in intravesical solution samples was 0.05%～10%, and the linear range for valrubicin assaying was 0.01～1.18 mg/ml. The LOD of the related substances of valrubicin in simulated samples spiked with the blank excipient was 0.1 μg/ml. The average recovery of the method was 100.5% with RSD of 0.4% (n=9); and the average content 96.8% was obtained from the precision test with RSD of 0.3% (n=9). Conclusion The method showed much higher chromatographic performance and specificity than the USP monographic method. It′s sensitive, precise, accurate and robust for assay and