RP-HPLC法测定安塞曲匹的有关物质

目的：建立RP-HPLC法测定安塞曲匹有关物质。方法：采用Agilent Zorbax SB C₁₈（150 mm×4.6 mm，5μm）色谱柱，流动相采用乙腈-0.1%磷酸溶液，梯度洗脱，流速1.0 mL·min^-1，柱温30℃，检测波长210 nm。结果：在选定的色谱条件下，主峰与各杂质峰均能良好分离，杂质A、杂质B、杂质C、杂质D、杂质E和安塞曲匹分别在0.41~24.52 μg·mL^-1（r=0.999 9）、0.50~29.93 μg·mL^-1（r=0.999 9）、0.46~27.88μg·mL^-1（r=0.999 6）、0.58~34.99μg·mL^-1（r=0.999 8）、0.51~30.88μg·mL^-1（r=0.999 8）、0.55~1 090.4μg·mL^-1（r=0.999 9）浓度范围内与峰面积呈良好线性关系。上述各杂质的定量下限分别为4.95、3.23、5.02、5.44、3.79、3.37 ng，平均回收率（n=9）分别为93.5%、92.5%、94.1%、91.0%、93.2%。结论：经方法学验证，本法可用于安塞曲匹的有关物质测定。

Objective: To establish an RP-HPLC method for the determination of related substances of anacetrapib. Methods: The determination was performed on an Agilent Zorbax SB C₁₈ column(150 mm×4.6 mm, 5μm),with a mobile phase consisting of a mixture of acetonitrile and 0.1% phosphoric acid solution by gradient elution. The flow rate was 1.0 mL·min^-1. The column temperature was set at 30℃ and the detective wavelength was 210 nm. Results: Related substances were completely separated from the principal component. The calibration curves for impurity A,impurity B,impurity C,impurity D,impurity E and anacetrapib,revealed good linearities over the ranges of 0.41-24.52μg·mL^-1 (r=0.999 9),0.50-29.93μg·mL^-1 (r=0.999 9),0.46-27.88μg·mL^-1 (r=0.999 6),0.58-34.99μg·mL^-1 (r=0.999 8),0.51-30.88μg·mL^-1 (r=0.999 8),0.55-1 090.4μg·mL^-1 (r=0.999 9),respectively. The limits of quantitation were 4.95, 3.23, 5.02, 5.44, 3.79, 3.37 ng, and the recoveries(n=9) were 93.5%, 92.5%, 94.1%, 91.0%, 93.2% for the above impurities, respectively. Conclusion: The method is proved by the methodology validation that it can be used for testing the related substances of anacetrapib.

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