Quality Study Of Anticancer Drug Cyclophosphamide

Cyclophosphamide is one of alkylating agents and commonly used as anti-tumor drugs.Which has good therapeutic effect on a variety of tumors in clinic.This study focuses on the quality study of cyclophosphamide,mainly including four parts:PartⅠ:A HPLC method was established for the determination of cyclophosphamide.By optimizing chromatographic conditions,the determination of cyclophosphamide was performed in an Alltima C₁₈ capillary column（4.6 mm×250 mm×5μm）.The results showed that the linearity of cyclophosphamide in the range of 234.472～937.889μg/m L was good（R²=0.99995）.The average recovery of cyclophosphamide was 100.3%with RSD（n=9）equal to 0.29%.The method has been proved to be suitable for the determination of cyclophosphamide content with wide linear range,short analysis time,good repeatability,good stability and high recovery.Part II:A headspace gas chromatography method was established for the determination of residual solvents such as acetone,dichloromethane and pyridine in cyclophosphamide.By optimizing chromatographic conditions,the detection of these three residual solvents was performed in a PEG-20M capillary column（30 m×0.32 mm×1.8μm）.The results showed that the linearities of the three residual solvents in the test concentration ranges are good.The quantification limits of the three residual solvent were less than or equal to 3.998μg/m L.Their detection limits were less than or equal to 1.120μg/m L.The average recoveries of acetone,dichloromethane and pyridine were 101.3%,86.4%and 107.0%with RSD（n=9）no more than1.5%.The proposed method was applied to detect three batchs of cyclophosphamide samples,dichloromethane and pyridine were not detected,but the detection range of acetone was 0.0100%～0.0114%.With high sensitivity,good specificity and accuracy,the method established in this work is suitable for simultaneous determination of the three residual solvent in cyclophosphamide.PartⅢ:A GC method was established for the determination of related substances1-amino-2-propanol（B）,2-amino-1-propanol（C）,N-methyl-2-pyrrolidone（D）,and3-hydroxypropanitrile（E）in intermediate 3-amino-propanol of cyclophosphamide for the first time.By optimizing chromatographic conditions,the separation of these four related substances was performed in a TG-WAXMS B capillary column（30 m×0.32 mm×0.25μm）.The results showed that the linearities of the four related substances in the test concentration ranges are good.The quantification limits of the four related substances were less than or equal to 7.999μg/m L.Their detection limits were less than or equal to 2.400μg/m L.The average recoveries of B,C,D and E were 108.7%,116.6%,97.1%and 89.2%with RSD（n=9）no more than 6.3%.The proposed method was applied to detect three batchs of3-amino-propanol samples,and B,D and E were not detected,but the detection range of C was 0.01%～0.02%.With high sensitivity,good specificity and accuracy,the method established in this work is suitable for the simultaneous determination of the four related substances in cyclophosphamide intermediate 3-amino-propanol.Part III:A GC method was established for the determination of related substances monoethanolamine（G）and triethanolamine（H）in intermediate diethanolamine of cyclophosphamide.By optimizing chromatographic conditions,the separation of these two related substances was performed in a CP-Volamine capillary column（30 m×0.32 mm×5μm）.The results showed that the linearities of the two related substances in the test concentration ranges are good.The quantification limits of the two related substances were less than or equal to 10.362μg/m L.Their detection limits were less than or equal to 3.109μg/m L.The average recoveries of G and H were 92.8%and 97.8%with RSD（n=9）no more than 5.5%.The proposed method was applied to detect three batchs of diethanolamine samples,and the detection range of G and H were 0.06%～0.09%,0.16%～0.22%.The established analytical method is simple,sensitive and applicable,and can be used to determine these two related substances in cyclophosphamide intermediate diethanolamine in a short time,which is helpful to improve the quality of cyclophosphamide.The thesis has 23 figures,49 tables and 65 references.