超高效液相色谱法测定盐酸博来霉素的有关物质

目的: 建立超高效液相色谱梯度洗脱法测定盐酸博来霉素中的有关物质。方法: 采用Waters HSS T3(100 mm×2.1 mm,1.8 μm)色谱柱,以己烷磺酸钠溶液(取己烷磺酸钠4.7 g与乙二胺四乙酸1.86 g,以0.08 mol·L^-1乙酸溶液溶解并稀释至1 000 mL 用氨溶液调节pH为4.3)为流动相A,甲醇-乙腈(70: 30)为流动相B,线性梯度洗脱,流速0.4 mL·min^-1,检测波长254 nm。结果: 博来霉素A₂质量浓度在4.75~237.4 μg·mL^-1范围内,与相应的峰面积呈良好的线性关系,r=0.999 6;博来霉素B₂质量浓度在2.23~111.6 μg·mL^-1范围内,与相应的峰面积呈良好的线性关系,r=0.999 6。方法的专属性、重复性及供试品溶液的稳定性良好。3批样品的最大单一杂质含量在1.2%~1.7%,总杂质含量在4.9%~6.5%。结论: 本法经方法学验证,可作为测定盐酸博来霉素组分与有关物质的有效方法。

Objective: To develop a gradient UPLC method for determination of the related substances of bleomycin hydrochloride. Methods: The Waters HSS T3 column(100 mm×2.1 mm,1.8 μm) was adopted.The mobile phase consisted of sodium hexane sulfonate and EDTA-2Na(pH 4.3,adjusted with ammonia) as the mobile phase A and methanol-acetonitrile(70: 30) as the mobile phase B.The linear gradient elution was used with the flow rate of 0.4 mL·min^-1 and the detection wavelength of 254 nm. Results: The good linear ranges of bleomycin A₂ and bleomycin B₂ were 4.75-237.4 μg·mL^-1 (r=0.9996) and 2.23-111.6 μg·mL^-1 (r=0.9996),respectively.This method had fine repeatability,specificity and the test solution had good stability.The content of the maximum single impurity of the samples was in the range of 1.2%-1.7%.The content of the total impurities of the samples was in the range of 4.9%-6.5%. Conclusion: By methodology validation,this method is proved to be suitable for determination of the components and the related substances of bleomycin hydrochloride.

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