摘要
建立了高效液相色谱(HPLC)-蒸发光散射(ELSD)法和HPLC-柱后衍生化荧光(FLD)法检测硫酸卡那霉素注射液中的有关物质。HPLC-ELSD法采用Welch LP-C18柱,以0.2 mol/L三氟乙酸溶液∶甲醇(98∶2)为流动相;HPLCFLD法采用Zorbax Eclipse Plus C18柱,以不同比例庚烷磺酸钠溶液与甲醇组成的流动相体系线性梯度洗脱,衍生化反应温度35℃,荧光激发波长340 nm,发射波长455 nm。结果显示,HPLC-ELSD法中各工艺杂质、主要降解杂质与卡那霉素及注射液中其他辅料之间分离度均良好,方法检测限为0.6µg/ml,含量为0.3%的最大未知杂质重复测定结果的RSD小于6%。HPLC-FLD法的检测限为0.03µg/ml,含量为0.4%的最大未知杂质重复测定结果的RSD小于1%。2个方法对于硫酸卡那霉素注射液的质量评价各有特点,优化后的HPLC-ELSD法分析速度快、检验成本低,适用于快速质量分析和评价;HPLC-FLD法的灵敏度和降解产物之间的分离度优于HPLC-ELSD法,适用于微量杂质的准确定量。
The high performance liquid chromatography(HPLC)-evaporative light scattering detection(ELSD)method and HPLC-fluorescence detection(FLD)with post-column derivatization method were established to detect the related substances in kanamycin sulfate injection.A Welch LP-C18 column was used for HPLC-ELSD method with the mobile phase of 0.2 mol/L trifluoroacetic acid solution∶methanol(98∶2).A Zorbax Eclipse Plus C18 column was used for HPLC-FLD method,and a linear gradient elution program was employed with the mobile phase composed of different ratios of sodium heptanesulfonate solution to methanol.The derivatization temperature was 35℃,the fluorescence excitation wavelength was 340 nm,and the emission wavelength was 455 nm.The results showed that for HPLC-ELSD method,the separation effects of the kanamycin process impurities,main degradation impurities and other excipients in injections were good.The detection limit of the method was 0.6µg/ml,and the RSD of repeated determination results of the largest unknown impurity with content of 0.3%was less than 6%.The detection limit of the HPLC-FLD method was 0.03µg/ml,and the RSD of repeated determination results of the largest unknown impurity with content of 0.4%was less than 1%.Two methods have their own characteristics for the quality evaluation of kanamycin sulfate injection.The optimized HPLC-ELSD method has fast analysis speed and low inspection cost,and it is suitable for the rapid quality analysis and evaluation.The sensitivity of HPLC-FLD method and the resolution between degradation products are better than those of HPLC-ELSD method,so that it is suitable for the accurate quantification of trace impurities.
作者
赵敬丹
刘浩
Zhao Jingdan;Liu Hao(Shanghai Institute for Food and Drug Control,Shanghai 201203)
出处
《中国医药工业杂志》
CAS
CSCD
北大核心
2020年第10期1311-1316,共6页
Chinese Journal of Pharmaceuticals
基金
2019年国家药品标准提高专项(2019H02)。
关键词
硫酸卡那霉素注射液
有关物质
柱后衍生化
蒸发光散射检测
荧光检测
kanamycin sulfate injection
relative substance
post-column derivatization
evaporative light scattering detection
fluorescence detection
