HPLC法测定富马酸伏诺拉生注射液中有关物质的含量

HPLC determination of related substances in vonoprazan fumarate injection

目的建立HPLC测定富马酸伏诺拉生注射液中有关物质含量的方法。方法采用YMC-Pack Pro C18色谱柱(4.6 mm×150 mm,3μm),流动相A为0.04 mol·L^(-1)磷酸二氢钾溶液(含0.2%三乙胺,用磷酸调节pH至6.6)-乙腈-异丙醇(92∶4∶4),流动相B为0.04 mol·L^(-1)磷酸二氢钾溶液(含0.2%三乙胺,用磷酸调节pH至6.6)-乙腈(40∶60),采用梯度洗脱,流速1.0 mL·min^(-1),柱温40℃,检测波长220 nm,进样量20μL。结果杂质H、杂质B、杂质C、杂质F、杂质D、杂质E、杂质J和伏诺拉生的质量浓度分别在0.12~4.99、0.12~4.99、0.12~4.90、0.13~5.01、0.12~4.99、0.13~5.01、0.13~5.01、0.13~5.01μg·mL^(-1)内与峰面积呈良好线性关系(r>0.9990);7个杂质的回收率为94.72%~100.67%,其RSD值均小于2.0%;7个杂质和伏诺拉生的定量限为0.12~0.13μg·mL^(-1),检测限为0.040~0.043μg·mL^(-1)。结论经方法学验证,本法专属性强、灵敏度高、准确度高,可用于富马酸伏诺拉生注射液中有关物质的含量测定。

Objective To establish an HPLC method to determine the related substances in vonoprazan fumarate injection.Methods YMC-Pack Pro C18 column(4.6 mm×150 mm,3μm)was used.Mobile phase A was 0.04 mol·L^(-1)potassium dihydrogen phosphate solution(containing 0.2%triethylamine,adjusted to pH 6.6 with phosphoric acid)-acetonitrile-isopropanol(92∶4∶4),while mobile phase B was 0.04 mol·L^(-1)potassium dihydrogen phosphate solution(containing 0.2%triethylamine,adjusted to pH 6.6 with phosphoric acid)-acetonitrile(40∶60).Gradient elution was used,the flow rate was 1.0 mL·min^(-1),the column temperature was 40℃,the detection wavelength was 220 nm,and the injection volume was 20μL.Results Impurity H,impurity B,impurity C,impurity F,impurity D,impurity E,impurity J and vonoprazan all showed good linear relationship with the peak area at the concentration ranges of 0.12~4.99,0.12~4.99,0.12~4.90,0.13~5.01,0.12~4.99,0.13~5.01,0.13~5.01 and 0.13~5.01μg·mL^(-1)(r>0.9990).The average recoveries of the above 7 impurities ranged 94.72%~100.67%,with all the RSD less than 2.0%.The quantitation limits of 7 impurities and vonoprazan were 0.12~0.13μg·mL^(-1)and the detection limits were 0.040~0.043μg·mL^(-1).Conclusion The method is exclusive,sensitive and accurate,which can be used to determine the related substances in vonoprazan fumarate injection.