HPLC法测定米力农注射液有关物质

目的：建立HPLC法测定米力农注射液有关物质，控制产品质量。方法：色谱柱为Venusil MP C₁₈（4.6 mm×250 mm，5 μm），柱温35 ℃；流动相为水-甲醇-硼酸钠缓冲液（725：250：25），流速1 mL·min^-1；检测波长254 nm。结果：米力农峰与各杂质峰间的分离度良好；米力农、米力农杂质A和米力农杂质B浓度分别在0.5~6.0 μg·mL^-1（r=1.000）、0.25~3.0 μg·mL^-1（r=0.9998）和0.25~3.0 μg·mL^-1（r=0.9998）内线性关系良好；杂质A和杂质B的平均回收率分别为99.1%（RSD=0.5%，n=3）和98.8%（RSD=0.9%，n=3）；米力农、杂质A和杂质B的定量限分别为0.17、0.17和0.7 ng。结论：建立的方法符合方法学验证要求可用来测定米力农注射液的有关物质，经对不同批号和同一批号不同流通区域抽样产品的检验，其有关物质的量与储存时间和区域分布呈明显的相关性。

Objective: To establish an HPLC method for the determination of related substances to control the quality of milrinone injection. Methods: A Venusil MP C₁₈ column (4.6 mm×250 mm,5 μm) was adopted and the column temperature was set at 35 ℃,the mixture of water-methanol-sodium borate buffer (725:250:25) was used as the mobile phase and the flow rate was 1 mL·min^-1,and the detection wavelength was 254 nm. Results: Milrinone and its related substances were separated well,and the calibration curves were linear within the concentration range of 0.5-6.0 μg·mL^-1(r=1.000)for milrinone,0.25-3.0 μg·mL^-1(r=0.9998) for milrinone impurity A,and 0.25-3.0 μg·mL^-1(r=0.9998) for milrinone impurity B,respectively.The average recoveries for milrinone impurity A and milrinone impurity B were 99.1% (RSD=0.5%,n=3) and 98.8% (RSD=0.9%,n=3),respectively.The limit of quantification(LOQ) of milrinone,milrinone impurity A and milrinone impurity B were 0.17,0.17 and 0.7 ng,respectively. Conclusion: The method meets the requirement of methodology validation.The analysis results of milrinone injection showed that there is an evident correlation between the amount of related substances,the storage time,and the regional distribution of the injection.

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