注射用头孢唑肟钠有关物质测定方法的建立

Establishment of an HPLC method for determination of related substance of cefazoxime sodium for injection

目的建立注射用头孢唑肟钠有关物质HPLC分析方法并对国产样品杂质情况行分析。方法采用Agilent Zorbax SB-C_(18)(250 mm×4.6 mm,5μm)色谱柱,柱温:35℃,流动相:以pH3.6乙酸铵溶液为流动相A,乙腈为流动相B,线性梯度洗脱:0→10 min,97%A→90%A;10→20 min,90%A→80%A;20→30 min,80%A;30→35 min,80%A→15%A;35→45 min,15%A;45→46 min,15%A→97%A,46→55 min,97%A;流速:0.8 mL/min,检测波长:254 nm,进样量:20μL。结果酸、碱、氧化、高温、光照强制降解试验证明方法的专属性良好;头孢唑肟及头孢唑肟诸分别在0.75~4.5μg/mL浓度范围内线性关系良好;头孢唑肟及10种已知杂质(杂质18、开环脱羧S-氧化物1、开环脱羧S-氧化物2、杂质KTO、杂质16、杂质6、杂质3、杂质13、二聚体及杂质12)的定量限分别为1.63、6.30、2.78、2.84、2.31、7.86、3.06、1.62、1.90、3.87和3.09 ng;各已知杂质平均回收率均在“96.5%~101.6%”之间;重复性RSD均符合要求。结论所建立的方法专属性、准确性及耐用性均良好,可用于注射用头孢唑肟钠有关物质的检测。

Objective This study constructed an HPLC method for the determination of the related substance cefazoxime sodium for injection and analyzed the impurities in domestic products.Methods The HPLC analysis was performed on an Agilent Zorbax SB-C_(18)column(250 mm×4.6 mm,5μm)with the column temperature at 35 o C.The mobile phase A was the pH 3.6 ammonium acetate solution,and the mobile phase B was acetonitrile.Gradient elution was performed as follows:0→10 min,97%A→90%A;10→20 min,90%A→80%A;20→30 min,80%A;30→35 min,80%A→15%A;35→45 min,15%A;45→46min,15%A→97%A;46→55min,97%A.The flow rate was 0.8 mL/min.The detection wavelength was 254 nm.The injection volume was 20μL.Results The acid,alkali,oxidation,high temperature,and light-forced degradation tests demonstrated good specificity of the methods.The linear range of cefzoloxime and cefazoxime was found to be between 0.75~4.5μg/mL with a good linear relationship.The quantification limits of cefotaxime and 10 of the known impurities(impurity 18,open ring decarboxylated S-oxide1,open ring decarboxylated S-oxide2,impurity KTO,cefzoloxime open ring,impurity D,impurity 3,cefzoloxime E isomer,dimer and impurity 12)were 1.63,6.30,2.78,2.84,2.31,7.86,3.06,1.62,1.90,3.87 and 3.09 ng,respectively.The average recoveries of all known impurities were between 96.5%and 101.6%.The RSD of repeatability all met the requirements.Conclusion The method proved to be specific,accurate and rugged,which made it suitable for the related substance analysis of cefazoxime sodium for injection.It could be used for the control of cefazoxime sodium for injection in the domestic market.