高效凝胶色谱法测定头孢替唑钠及其注射剂中的聚合物类杂质

目的 建立高效凝胶色谱法测定头孢替唑钠及其注射剂中的聚合物杂质，并采用二维液质联用法对聚合物杂质的结构进行推断。方法 采用TSK-Gel 2000SWXL凝胶色谱柱(7.8mm×300mm, 5μm)，0.005mol/L磷酸盐缓冲液0.005mol/L磷酸氢二钠-0.005mol/L磷酸二氢钠(61:39)]-乙腈(95:5)为流动相，流速为0.7mL/min，检测波长254nm，进样量20μL；二维色谱采用Waters HSS T3(100mm×2.1mm, 1.8μm)，以0.1%甲酸水溶液为流动相A，乙腈为流动相B，梯度洗脱；质谱采用ESI离子源，正离子模式检测。结果 主成分前后共分离得到6个杂质峰；头孢替唑检测浓度的线性范围为0.5~20μg/mL(r=1.0000)，检测限与定量限分别为0.015和0.05μg/mL，重复性试验各杂质含量的RSD均在合理范围内(n=6)；采用LC-MS推定了3个聚合物类杂质的结构。结论 本方法灵敏度和准确度好，适用于头孢替唑钠及其制剂中聚合物杂质的控制。

Determination of polymer impurities in ceftezole sodium and ceftezole sodium for injection by HPGFC

Objective To establish a high performance gel filtration chromatography (HPGFC) method for the separation and analysis of polymers in ceftezole sodium and ceftezole sodium for injection, and determine the structures of the impurities by 2D-LC-MS. Methods HPGFC was performed by using TSK-Gel 2000SWXL (7.8mm×300mm, 5μm) column. The mobile phase was phosphate buffer solution 0.005mol/L disodium hydrogen phosphate buffer solution-0.005mol/L sodium dihydrogen phosphate buffer solution (61:39)]-acetonitrile (95:5). The flow rate was 0.7mL/min, the detection wavelength was 254nm，and the injection volume was 20μL. Two-dimensional chromatographic conditions: Waters HSS T3 column(50mm×2.1mm, 1.8μm) was adopted, with the solution containing 0.1% formic acid as mobile phase A and acetonitrile as mobile phase B. Gradient elution was used. Electrospray ionization (ESI) source was used at the positive ion detection mode. Results Six impurity peaks were obtained in total. The linear range of ceftezole was 0.5~20μg/mL(r=1.0000). The limit of detection was 0.015μg/mL and the limit of quantitation was 0.05μg/mL. The RSD of repeatability was qualified (n=6). Three polymers were identified by LC-MS. Conclusion The HPGFC method can be used for the quantitative and qualitative analysis of polymer impurities. It is sensitive and accurate for the control of polymers in ceftezole.