甘露醇原料药及其制剂中甘露醇的高效液相色谱-电喷雾检测器分析方法

Determination of Mannitol in Mannitol API and Its Preparation by HPLC-CAD

目的建立甘露醇原料药及其制剂中甘露醇的高效液相色谱-电喷雾检测器(HPLC-CAD)分析方法。方法色谱柱为Waters XBridge Amide柱(250 mm×4.6 mm,3.5μm),流动相为乙腈-水(70∶30,V/V),流速为1.0 mL/min,柱温为35℃,进样量为10μL;雾化温度为55℃,幂率为1.0,采样频率为10 Hz,过滤常数为3.6 s。结果甘露醇质量浓度在10~200μg/mL范围内与峰面积线性关系良好(r=0.9978,n=5);平均回收率为99.96%,RSD为1.24%(n=9);定量限为1.0μg/mL,检测限为0.2μg/mL;精密度、稳定性、重复性试验的RSD均小于2.0%。3批甘露醇原料药中甘露醇的含量测定结果分别为99.21%,99.43%,99.32%,RSD均小于1.0%(n=4);有关物质总杂质含量分别为0.25%,0.21%,0.23%。3批注射用胸腺五肽中甘露醇的含量测定结果分别为0.94,0.93,0.91 mg/mg,RSD均小于1.50%(n=4)。结论该方法简便、快速、灵敏、准确,可用于甘露醇原料药及其制剂中甘露醇的含量测定及质量控制。

Objective To establish a high-performance liquid chromatography-charged aerosol detection(HPLC-CAD)method for the determination of mannitol in mannitol active pharmaceutical ingredients(API)and its preparation.Methods The chromatographic column was Waters XBridge Amide column(250 mm×4.6 mm,3.5μm),the mobile phase was acetonitrile-water(70∶30,V/V),the flow rate was 1.0 mL/min,the column temperature was 35℃and the injection volume was 10μL.The detector of CAD was applied with nebulization temperature at 55℃,power function value(PFV)at 1.0,collection frequency at 10 Hz and filter constant at 3.6 s.Results The linear range of mannitol was 10-200μg/mL(r=0.9978,n=5).The average recovery of mannitol was 99.96%with an RSD of 1.24%(n=9).The limit of quantification(LOQ)was 1.0μg/mL and the limit of detection(LOD)was 0.2μg/mL.The RSDs of precision,stability and repeatability tests were less than 2.0%.The contents of mannitol in the three batches of mannitol API were 99.21%,99.43%and 99.32%with RSDs less than 1.0%(n=4),respectively,and the total impurity contents of related substances in the above three batches of mannitol API were 0.25%,0.21%and 0.23%,respectively.The contents of mannitol in the three batches of Thymopentin for Injection were 0.94,0.93 and 0.91 mg/mg with RSDs less than 1.50%(n=4),respectively.Conclusion The method was simple,rapid,sensitive and accurate,which can be used for the determination and quality control of mannitol in mannitol API and its preparation.