梯度洗脱HPLC测定奥达特罗的有关物质

Detection of related substances of olodaterol by HPLC with gradient elution

目的:建立奥达特罗有关物质检查的方法。方法:采用Sapphire C18色谱柱(250 mm×4.6 mm,5μm);以10 mmol·L-1磷酸二氢钠溶液为流动相A、乙腈为流动相B进行梯度洗脱;流速为1 m L·min-1;检测波长为220 nm;柱温30℃。结果:主成分与5种有关物质分离完全。奥达特罗的线性范围为0.01~25μg·mL^(-1),相关系数r为1.000 0,检测限为4.06 ng·mL^(-1)。各杂质的线性范围为0.04~5μg·mL^(-1),相关系数r≥0.999 9,精密度良好。奥达特罗1%自身对照溶液重复进样的RSD值为0.35%。各已知杂质以杂质对照品法,未知杂质以主成分自身对照法计算杂质含量,3批样品最大单个杂质不超过0.3%,总杂质含量不超过0.7%。结论:该方法简便、灵敏、准确、专属性强,可用于奥达特罗有关物质的检查。

Objective： To establish an HPLC method for the detection of related substances of olodaterol.Methods： The separation was performed on Sapphire C18 column（ 250 mm × 4. 6 mm,5 μm）. Gradient elution was used to determine the related substances of olodaterol with 10 mmol·L-1 sodium dihydrogen phosphate as mobile phase A and acetonitrile as mobile phase B. The flow rate was 1 m L·min-1. The detection wavelength was220 nm,and the column temperature was 30 ℃. The contents of known impurities was determined by external standard method and that of the unknown impurities were calculated by main component self-comparison method.Results： The related substances were completely separated from the principal component. The linear range of olodaterol was 0. 01 - 25 μg·m L^-1,with correlation coefficient of 1. 000 0 and limit of detection of 4. 06 ng·m L^-1.The linear ranges of the impurities were 0. 04 - 5 μg·m L-1（ r≥0. 999 9）. The precision was good,with RSD of0. 35% for duplicate injection of 1% olodaterol solution. The content of individual impurity was lower than 0. 3%,and that of total impurities was lower than 0. 7%. Conclusion： The method is simple,sensitive,accurate and specific,and can be used to determine the related substances of olodaterol.