目的 采用超高效液相色谱质谱联用（UPLC-MS/MS）法同时测定异环磷酰胺原料药中主成分及2种有关物质。方法 采用ACQUITY UPLC^® HSS T3（100 mm×2.1 mm，1.8 μm）色谱柱，以甲醇（A）-5 mmol·L-1碳酸氢铵溶液（B）为流动相进行梯度洗脱，对异环磷酰胺原料药中的异环磷酰胺、杂质G、杂质E进行分离，体积流量为0.3 mL·min^-1，进样量为10 μL，柱温为30℃，质子离子源ESI，正离子模式下以多反应监测方式（MRM）进行定量分析。结果 异环磷酰胺、杂质E、杂质G的线性范围分别1.00～50.00、1.00～50.00、0.01～0.5 ng·mL^-1，线性关系良好，r均大于0.999 4；且该方法的仪器精密度、重复性良好，异环磷酰胺的回收率为98.1%～101.7%、RSD为0.40%～1.73%，杂质E的回收率为96.1%～111.1%、RSD为0.67%～2.34%，杂质G的回收率为93.7%～108.3%、RSD为2.2%～7.6%，符合标准。原料药中的异环磷酰胺质量分数为97.1～98.5%，杂质E均未检出，杂质G在1501批次检出量最高，为0.009 25%，杂质E和G未超千分之一限度。结论 建立的UPLC-MS/MS法灵敏度较高、重复性好，为异环磷酰胺原料药的质量控制提供了一种可行的分析方法，适用于该产品的质量控制。

Objective To use liquid chromatography-mass spectrometry (UPLC-MS/MS) to simultaneously determine the principal components and two related substances in ifosfamide API. Methods The ACQUITY UPLC^® HSS T3 (100 mm × 2.1 mm, 1.8 μm) column was used to perform gradient elution of methanol (A) -5 mmol·L^-1 ammonium bicarbonate (B) as the mobile phase, and ifosfamide, impurity G and impurity E in ifosfamide API were separated with a flow rate of 0.3 mL·min^-1, an injection volume of 10 μL, a column temperature of 30 ℃, and a proton ion source ESI. Quantification in positive ion mode in multi-reaction monitoring mode (MRM). Results The linear ranges of ifosfamide, impurity E and impurity G were 1.00-50.00、1.00-50.00、0.01-0.5 ng·mL^-1 respectively, with good linear relationships and correlation coefficients greater than 0.994, and the instrument precision and repeatability of the method were good. The recoveries of isocyclophosphamide were 98.1%-101.7% with RSD of 0.40%-1.73%. The recovery of impurity E was 96.1%-111.1% with RSD of 0.67%-2.34%. The recoveries of impurity G were 93.7%-108.3% with RSD of 2.2%-7.6%. The mass fraction of ifosfamide in the raw material was 97.1%-98.5%, and impurity E had not been detected. The detection amount of impurity G in batch 1501 is 0.009 25%, and impurities E and G do not exceed the limit of one thousandth. Conclusion The proposed method has high sensitivity and good repeatability, which provides a feasible analysis method for the quality control of ifosfamide API, which is suitable for the quality control of this product.

R932

江苏省产教融合一流课程《药物分析》建设项目；江苏海洋大学专业学位研究生课程案例库建设项目（YJSZY202102）；江苏海洋大学2023年一流课程立项；连云港市政策引导产学研项目（CYX2202）