HPLC法测定富马酸沃诺拉赞原料药中的有关物质

目的：建立富马酸沃诺拉赞原料药有关物质的HPLC测定方法。方法：采用Symmetry C₁₈色谱柱（4.6 mm×250 mm，5 μm），对降解杂质和工艺杂质进行定量分析，以0.02 mol·L^-1磷酸氢二钾（加入磷酸调pH至5.0）-乙腈为流动相，梯度洗脱，流速1.0 mL·min^-1，检测波长为235 nm，柱温30℃。用UPLC-MS鉴定其中1个未知杂质；采用ACQUITY UPLC BEH C₁₈色谱柱（50 mm×2.1 mm，1.7 μm），以乙腈为流动相A，0.1%乙酸+2 mmol·L^-1醋酸铵溶液为流动相B，梯度洗脱，ESI离子源，正离子扫描方式，检测离子m/z为361.9。结果：主峰与各杂质峰间能达到基线分离，沃诺拉赞及各杂质质量浓度在0.2~2.0 μg·mL^-1范围内与峰面积呈良好的线性关系（r=0.999），最低检测限为0.1 μg·mL^-1。3批样品有关物质测定结果显示，已知杂质和未知杂质含量均低于0.1%；其中1个未知杂质为1-［5-（2-氟苯基）-1-（N-氧化物吡啶-3-基-磺酰基）-1H-吡咯-3-基］-N-甲基甲胺。结论：本法可作为富马酸沃诺拉赞质量控制的方法。

Objective: To establish an HPLC method for the determination of the related substances in vonoprazan fumarate.Methods: The test for degradation and process impurities was performed on a symmetry C₁₈ column(4.6 mm×250 mm,5 μm),the mobile phase consisted of 0.02 mol·L^-1 potassium phosphate solution(adjusted to pH 5.0 with phosphoric acid)-acetonitrile with gradient elution at the flow rate of 1.0 mL·min^-1;the detection wavelength was 235 nm and the column temperature was 30℃.One of the unknown impurities was identified by UPLC-MS with the gradient elution.The separation was performed on the ACQUITY UPLC BEH C₁₈ column(50 mm×2.1 mm,1.7 μm),using acetonitrile as mobile phase A and 0.1% acetic acid+2 mmol·L^-1 ammonium acetate as the mobile phase B.The mass spectrometer was operated in positive electrospray ionization modeswith the transition m/z of 361.9.Results: The impurity peaks were separated well from the main peak.A good linearity was observed over the range of 0.2-2.0 μg·mL^-1(r=0.999),the LOD was 0.1 μg·mL^-1;the determination results of the three batches of samples showed that each of the impurities was less than 0.1%.The unknown impurity identified was 3-((2-(2-fluorophenyl)-4-((methylamino)methyl)-1H-pyrrol-1-yl)sulfonyl)pyridine-1-oxide.Conclusion: The established determination method can be used for quality control of vonoprazan fumarate.