[關(guān)鍵詞]
[摘要]
目的 建立氣相色譜(GC)法測定異環(huán)磷酰胺原料藥中3種有關(guān)物質(zhì)(起始物料3-氨基丙醇、雜質(zhì)D和雜質(zhì)F)。方法 采用HP-55% Phenyl Methyl Siloxane色譜柱(30 m×320 μm,0.25 μm);采用程序升溫的方式,起始柱溫50℃,保持2 min后以每分鐘5℃的速率升溫至180℃,再以每分鐘20℃的速率升溫至260℃;進樣口溫度為380℃;檢測器溫度為380℃;分流比5∶1。結(jié)果 起始物料峰、雜質(zhì)D、雜質(zhì)F與周圍峰分離度分別為2.6、3.0、1.6;異環(huán)磷酰胺起始物料3-氨基丙醇在39.80~199.0 μg·mL-1線性關(guān)系良好(r=0.999 1),雜質(zhì)D在39.48~197.4 μg·mL-1線性關(guān)系良好(r=0.999 1),雜質(zhì)F在2.000~20.000 μg·mL-1線性關(guān)系良好(r=0.999 9); 3-氨基丙醇和雜質(zhì)D的檢測限分別為9.95、9.87 μg·mL-1,雜質(zhì)F因色譜柱柱效降低且進樣量過大未能進行檢測限測定;3-氨基丙醇、雜質(zhì)D、雜質(zhì)F的定量限分別為19.90、19.74、2.000 μg·mL-1;起始物料、雜質(zhì)D和雜質(zhì)F回收率均在90.09%~115.90%,回收率RSD分別為4.8%、7.2%、2.3%。結(jié)論 該方法的專屬性、精密度、線性關(guān)系良好,準確可靠,符合異環(huán)磷酰胺中3種有關(guān)物質(zhì)的檢測要求。
[Key word]
[Abstract]
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.
[中圖分類號]
R932
[基金項目]
江蘇省產(chǎn)教融合一流課程《藥物分析》建設(shè)項目;江蘇海洋大學專業(yè)學位研究生課程案例庫建設(shè)項目(YJSZY202102);江蘇海洋大學2023年一流課程《藥物儀器分析》建設(shè)項目;連云港市政策引導產(chǎn)學研項目(CYX2202)