LC-MS/MS法分析注射用哌拉西林钠的杂质谱

目的: 建立LC-MS/MS 法分析注射用哌拉西林钠中的有关物质的杂质谱。方法: 采用Kromasil Eternity-5 C₁₈(2.1 mm×150 mm,3.5 μm)色谱柱,以0.2%甲酸溶液(A)-乙腈(B)为流动相,进行梯度洗脱,流速0.2 mL·min^-1;通过光电二极管阵列检测器(PDA)及电喷雾离子化串联质谱检测法,采集杂质谱的PDA数据、质谱的母离子及子离子数据,结合质谱的降解规律,杂质的色谱行为,相关对照品的裂解规律,以及主成分哌拉西林钠的合成与降解反应,进行未知杂质结构解析,并对该结构进行初步验证。结果: 在所建立的条件下,杂质谱中哌拉西林钠及其杂质分离良好,分离出5组同分异构体,共检测到16个较大杂质峰,其中,8个杂质峰为已知杂质峰,分别为BP杂质E、BP杂质A、BP杂质C、BP杂质C同分异构体、BP杂质B、BP杂质B同分异构体、BP杂质F、BP杂质D,8个未知杂质峰中,分别为哌拉西林2个异构体、BP杂质B降解物、哌拉西林去4-乙基-2,3-二氧代-1-哌嗪降解物、哌拉西林去4-乙基-2,3-二氧代-1-哌嗪降解物异构体、BP杂质A与B二聚体、BP杂质A与B二聚体的异构体、哌拉西林二聚体。结论: 建立的LC-MS/MS法能有效地洗脱、分离注射用哌拉西林钠的杂质,并分析其杂质谱,为哌拉西林钠的原料和相关制剂质量控制和工艺优化提供了参考依据。

Objective: To establish an LC-MS/MS method for the identification and determination of the impurity spectra in piperacillin sodium for injection. Methods: The HPLC separation was carried out on a Kromasil Eternity-5 C₁₈(2.1 mm×150 mm,3.5 μm)column by gradient elution,with a mobile phase consisting of 0.2% formic acid aqueous solution(A)and acetonitrile(B)at a flow rate of 0.2 mL·min^-1.The results were obtained by ESI-MS and tandem mass spectrometry.The PDA,parent ions and the corresponding product spectra of all impurity profiles in piperacillin sodium for injection were determined and elucidated.According to fragmentation regularity of mass spectrometry,chromatographic behavior of impurity,fragmentation regularity of related reference substances,as well as synthesis and degradation of the principal component,piperacillin sodium,the structures of each impurity were resolved,speculated and determined primarily. Results: Good resolution of piperacillin sodium and the main related substances was achieved and five groups of isomers were obtained.Among sixteen relatively great impurity peaks,8 peaks were known,including BP impurity E,BP impurity A,BP impurity C,isomeride of BP impurity C,BP impurity B,isomeride of BP impurity B,BP impurity F,BP impurity D.The other 8 peaks were unknown,including 2 isomers of piperacillin,the degaration of BP impurity B,piperacillin 4-ethyl-2,3-oxo-piperazine degradation products,piperacillin 4-ethyl isomer-2,3-oxo-piperazine degradation products,the dipolymer of BP impurity B and A,the isomeride of dipolymer of BP impurity B and A,and the dipolymer of piperacillin. Conclusion: The established method is effective for the separation and identification of the related substances in piperacillin sodium for injection.Our results provide a useful reference for the quality control of its raw materials and the related preparations as well as for process optimization.