头孢他啶杂质H研究

Study on ceftazidime impurity H

目的:对国产头孢他啶中共有的杂质H进行研究。方法:制备液相色谱采用Phenomenex Luna C18柱(150 mm×21.2 mm,5μm),流动相A为醋酸盐缓冲液(20 mmol·L-1醋酸铵,用醋酸调节pH为4.0),流动相B为乙腈,梯度洗脱,流速15 mL·min-1,柱温为室温。收集目标物冻干后利用紫外光谱、红外光谱、质谱、核磁共振对头孢他啶杂质H进行结构确认;并利用斑马鱼模式生物比较头孢他啶及其杂质H的胚胎发育毒性。结果:通过实验证实头孢他啶杂质H的结构为(6R,7R)-7-[[(2-氨基-1,3,4-噻唑-4-基)-2-(1-甲氧基2-甲基-1-氧代内烷-2-基)氧亚氨乙酰]氨基]-8-氧代-3-(吡啶-1-鎓-1-基-甲基)-5-硫杂-1-氮杂双环[4.2.0]辛-2-烯-2-甲酸;头孢他啶杂质H在斑马鱼胚胎毒性实验中的致畸作用是头孢他啶的25倍、致死作用是头孢他啶的8倍。结论:头孢他啶杂质H应予以严格控制。

Objective：To study the common impurity H in domestic ceftazidime. Methods：The HPLC system consisted of a Phenomenex Luna C18column（ 150 mm × 21. 2 mm,5 μm）,the mobile phase A was acetate buffer solution（ 20 mmol·L1ammonium acetate,pH adjusted to 4. 0 with acetic acid）,the mobile phase B was acetonitrile forgradient elution with the UV detector at 254 nm,the flow rate at 15 mL·min1,the column temperature at room temperature. The object was collected and freezedried. Ultraviolet spectrum,infrared spectrometry,mass spectrometry and nuclear magnetic resonance were adopted to confirm the chemical structure of ceftazidime impurity H. Thezebrafish was used as an ideal model to compare teratogenesis and toxicity between ceftazidime and impurity H.Results：The chemical structure of ceftazidime impurity H was confirmed to be（ 6R,7R）7[[（ 2Z）2-（ 2-aminothiazol4yl）2-（ 2methoxy1,1dimethyl2oxoethoxy） imino]acetyl]amino]8oxo3-[（ 1-pyridinio） methyl]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate. Ceftazidime impurity H was 25 times higher in teratogenesis and 8times higher in toxicity compared with ceftazidime in zebrafishembryo toxicity test. Conclusion：Ceftazidime impurity H should be controlled strictly.