利伐沙班中苯胺类潜在基因毒性杂质的HPLC法测定

目的：建立高效液相色谱法测定利伐沙班中苯胺类潜在基因毒性杂质4-（4-氨基苯基）-3-吗啉酮。方法：采用Hedera^TM苯基硅烷键合相色谱柱（250 mm×4.6 mm，5 μm），流动相A为乙腈，流动相B为乙腈-10 mmol·L^-1磷酸二氢钠溶液（pH 3.5）（10：90），梯度洗脱，流速1.2 mL·min^-1，柱温30℃，检测波长242 nm。结果：杂质4-（4-氨基苯基）-3-吗啉酮质量浓度在20.12~201.2 ng·mL^-1范围内线性关系良好（r=0.998 1），检测限为10.10 ng·mL^-1，定量限为30.30 ng·mL^-1，平均回收率为97.4%（RSD=2.8%，n=9），重复性良好（RSD=2.0%，n=6）。3批原料药中未检出4-（4-氨基苯基）-3-吗啉酮。结论：经方法学验证，本法适用于利伐沙班中4-（4-氨基苯基）-3-吗啉酮的测定。

Objective: To develop an HPLC method for the determination of the aniline potentially genotoxic impurity,4-4-(aminophenyl)-3-morpholinone,in rivaroxaban.Methods: A Hedera^TM phenyl column (250 mm×4.6 mm,5 μm)was adopted with a gradient elution system.The mobile phase consisted of acetonitrile(A phase)and acetonitrile-10 mmol·L^-1 sodium dihydrogen phosphate aqueous solution(pH 3.5) (10:90,B phase).The flow rate was 1.2 mL·min^-1,and the column temperature was 30℃.The ultraviolet detection wavelength was 242 nm.Results: The calibration curve was linear for 4-(4-aminophenyl)-3-morpholinone in the range of 20.12-201.2 ng·mL^-1(r=0.998 1).The limit of detection and limit of quantification were 10.10 ng·mL^-1 and 30.30 ng·mL^-1,respectively.The average recovery was 97.4%,with RSD of 2.8%(n=9).The repeatability was fine,with RSD of 2.0%(n=6).The impurity was not detected in three batches of rivaroxaban.Conclusion: The method was applicable for the quality control of 4-(4-aminophenyl)-3-morpholinone in rivaroxaban through the methodology validation.

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