硫普罗宁注射液有关物质的分离与测定

目的优化反相高效液相色谱法分离测定硫普罗宁注射液有关物质并进行方法学验证。方法以各杂质峰的对称性、相对保留时间及分离度等因素为指标,优化了pH、流动相缓冲盐浓度、温度等因素,建立HPLC色谱条件:色谱柱:Waters Symmetry-C18柱;流动相:0.05moL.L-1磷酸二氢钾溶液(用磷酸调pH值至3.2)-乙腈(95∶5);检测波长:210nm;柱温:30℃;流速:1.0mL.min-1。结果优化后的方法具有简便快速、灵敏度高、专属性强的特性;杂质A、B及杂质C、杂质D的相对响应因子(相对于硫普罗宁)分别为0.73、2.32和1.18;硫普罗宁注射液中含有这4个杂质,且含量均大于0.1%。结论采用加相对响应因子的1%主成分自身对照法以及杂质对照外标法更能准确反映杂质含量。

Determination of Related Substances in Tiopronin Injection by HPLC

Objective To optimize a HPLC method for the separation and determination of related substance in Tiopronin Injection.Methods Based on the evaluation results of the symmetry,relative retention time and resolution factors,chromatographic conditions were optimized.The analysis was conducted by using Waters Symmetry C18 column with a mobile phase of acetonitrile-0.05moL·L-1 phosphate buffer（adjust to pH 3.2 with phosphoric acid）（5∶95）.The flow rate was 1.0mL·min-1,and the detection wavelength was 210nm.Results The optimized method was developed with simple,time-saving,sensitive and specific characteristics.The correction factors of impurity A,B,impurity C and impurity D were 0.73,2.32 and 1.18 respectively.Four impurities,which contents were greater than 0.1%（w/w）,were detected in Tiopronin Injection.Conclusion External standard methods with correction factors is a better way to accurately reflect the content of the impurity.