目的 建立气相色谱（GC）法测定异环磷酰胺原料药中3种有关物质（起始物料3-氨基丙醇、杂质D和杂质F）。方法 采用HP-55% Phenyl Methyl Siloxane色谱柱（30 m×320 μm，0.25 μm）；采用程序升温的方式，起始柱温50℃，保持2 min后以每分钟5℃的速率升温至180℃，再以每分钟20℃的速率升温至260℃；进样口温度为380℃；检测器温度为380℃；分流比5∶1。结果 起始物料峰、杂质D、杂质F与周围峰分离度分别为2.6、3.0、1.6；异环磷酰胺起始物料3-氨基丙醇在39.80～199.0 μg·mL^-1线性关系良好（r=0.999 1），杂质D在39.48～197.4 μg·mL^-1线性关系良好（r=0.999 1），杂质F在2.000～20.000 μg·mL^-1线性关系良好（r=0.999 9）； 3-氨基丙醇和杂质D的检测限分别为9.95、9.87 μg·mL^-1，杂质F因色谱柱柱效降低且进样量过大未能进行检测限测定；3-氨基丙醇、杂质D、杂质F的定量限分别为19.90、19.74、2.000 μg·mL^-1；起始物料、杂质D和杂质F回收率均在90.09%～115.90%，回收率RSD分别为4.8%、7.2%、2.3%。结论 该方法的专属性、精密度、线性关系良好，准确可靠，符合异环磷酰胺中3种有关物质的检测要求。

Objective A gas chromatographic (GC) method was established for the determination of related substances (starting material 3-aminopropanol, impurity D and impurity F) in isocyclophosphamide API. Methods HP-5 5 % Phenyl Methyl Siloxane Column (30 m × 320 μm, 0.25 μm) was used. The column temperature was programmed to start at 50°C, held for 2 min and then ramped up to 180 ℃ at a rate of 5 ℃ per minute and then to 260 ℃ at a rate of 20 ℃ per minute; the inlet temperature was 380 ℃; the detector temperature was 380 ℃; and the split ratio was 5:1. Results Under the chromatographic conditions, three related substances in isocyclophosphamide API could be detected and the methodology was verified. The separation between the peak of the starting material and the surrounding peaks was 2.6, the separation between impurity D and the surrounding peaks was 3.0, and the separation between impurity F and the surrounding peaks was 1.6, with good separation effects all greater than 1.5. Starting materials, 3-aminopropanol, had a good linear relationship in the range of 39.80-199.0 μg·mL^-1 (r=0.999 1). Impurity D had a good linear relationship in the range of 39.48-197.4 μg·mL^-1 (r=0.999 1). Impurity F had a good linear relationship in the range of 2.000-20.00 μg·mL^-1 (r=0.999 9). The limits of detection (LODs) for 3-aminopropanol and impurity D were 9.95 and 9.87 μg·mL^-1, respectively, while the LOD for impurity F was not determined due to the reduced column efficiency and the large injection volume; the limits of quantification (LOQs) for 3-aminopropanol, impurity D, and impurity F were 19.90, 19.74, and 2.000 μg·mL^-1, respectively. The recovery rates of starting materials, impurity D and impurity F should be in the range of 80%-120%, and the recovery rate RSD should be less than 10%. Conclusion The method is characterised by good specificity, precision, linearity, accuracy and reliability, and is able to meet the requirements for the determination of the three substances of interest in isocyclophosphamide.

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《药物仪器分析》建设项目；连云港市政策引导产学研项目（CYX2202）