反相高效液相色谱法测定头孢地尼及制剂中的聚合物类杂质

Determination of polymer impurities in cefdinir and preparations by RP-HPLC

目的建立反相高效液相色谱法测定头孢地尼及制剂中的聚合物杂质。方法以CAPCELL PAK C_(18)MGⅡ为色谱柱(250 mm×4.6 mm,5μm),0.25%四甲基氢氧化铵溶液(p H 5.5)[每1000mL中加入0.1 mol/L乙二胺四乙酸二钠溶液0.4 mL]-甲醇-乙腈为流动相系统,梯度洗脱,建立了头孢地尼聚合物类杂质的分析方法并进行了方法学验证。采用二维液质联用法对检出的聚合物类杂质进行定性分析。结果在该色谱条件下头孢地尼及已知杂质均与头孢地尼聚合物间分离良好,头孢地尼检测浓度的线性范围为0.30~30.1μg/mL(r=1.0000),方法的检测限与定量限分别为1.5 ng和4.5 ng。所建立的方法能够检出头孢地尼二聚体,头孢地尼二聚体的多个同分异构体及头孢地尼三聚体。结论本方法专属性好,灵敏度高,耐用性好,可用于头孢地尼及其制剂中聚合物杂质的控制。

Objective To establish a reverse high-performance liquid chromatography(HPLC)for the separation and determination of polymers in cefdinir and its preparations.Methods The method was performed by using CAPCELL PAK C_(18)MGⅡcolumn(250 mm×4.6 mm,5μm).The mobile phase was 0.25%tetramethyl ammonium hydroxide(pH5.5)containing 0.1 mol/L disodium EDTA solution(0.4 mL per 1000 mL)-acetonitrilemethanol.The gradient elution was used.The method was validated,and the polymer impurities were analyzed by the two dimensional liquid chromatography-mass spectrometry method.Results Cefdinir,the identified impurities and polymers were separated effectively.The linear range of cefdinir was 0.30~30.1μg/mL(r=1.0000).The limit of detection was 1.5 ng and the limit of quantitation was 4.5 ng.Cefdinir dimer,isomers of cefdinir dimer,and cefdinir trimer can be detected by the new method.Conclusion The established RP-HPLC method with good robustness is specific and sensitive.Therefore,it can be used for the control of polymer impurities in the pharmaceutical preparations.