在线体积排阻色谱-反相液相色谱-飞行时间质谱法鉴定注射用头孢哌酮钠舒巴坦钠中聚合物杂质

Determination of the Polymer Impurities in Cefoperazone Sodium by Two-Dimensional SEC-RPLC-QTOFMS Method

目的建立在线体积排阻色谱-反相液相色谱-飞行时间质谱法(SEC-RPLC-QTOFMS)用于注射用头孢哌酮钠舒巴坦钠中聚合物杂质的鉴定。方法一维采用Shimpack Diol-150(7.9 mm×250 mm,5μm)色谱柱;进样量为20μL并以254 nm为检测波长,建立头孢哌酮和聚合物杂质的SEC分离条件;并将SEC分离所得聚合物杂质通过中心切割技术收集到二维RPLC中脱盐和进一步分离,采用Q-TOF为检测器,采集分离所得杂质一级和二级质谱信息后对其进行结构鉴定。结果在一维条件下头孢哌酮与聚合物峰分离良好;通过二维RPLC-QTOFMS推测出9个杂质的结构,其中有4个为闭环二聚物。结论在线SEC-RPLC-QTOFMS技术可以将聚合物指针性定位与在线脱盐高分辨质谱鉴定有机结合起来,为表征聚合物杂质的研究思路提供一定的参考。本实验建立的方法灵敏度高,适用于低含量杂质的结构鉴定,对β-内酰胺类抗生素中聚合物的分离、分析及产品质量提高提供了手段。

OBJECTIVE To establish a high performance size exclusion chromatography(SEC)method for determining the polymer impurities in cefoperazone sodium and its structure identification.METHODS The analysis was carried out by using a Shimpack Diol-150 column(7.9 mm×250 mm,5μm).The mobile phase was 95 volumes of NaH2PO4 and Na2HPO4 buffer and 5 volumes of acetonitrile.The detection wavelength was set at 254 nm.The injection volume was 20μL.The ion source was ESI ionization source,and the scan pattern was positive.RESULTS The polymers were completely separated from cefoperazone.The polymer impurities found by SEC mainly were degradation impurities and openβ-lactam ring impurities.CONCLUSION The method provides a basis for the separation and analysis ofβ-lactams polymers,and it can be used to improve the product quality.