HPLC梯度法测定注射用哌拉西林钠/他唑巴坦钠中2种成分含量

Determination of Two Components in Piperacillin Sodium/Tazobactam Sodium for Injection with HPLC Gradient Method

目的建立同时测定复方制剂注射用哌拉西林钠/他唑巴坦中钠中2种成分含量的方法。方法采用C18为固定相的色谱柱,流动相A为甲醇,流动相B为水-0.2 mol·L^(-1)磷酸二氢钠溶液-0.4 mol·L^(-1)四丁基氢氧化铵溶液(853∶143∶4)(用20%磷酸调节pH至4.5),采用梯度洗脱;检测波长为220 nm,流速为1.0 ml·min^(-1),进样量为10μl。结果能满足《中国药典》2020版含量测定的系统适用性要求;哌拉西林的线性范围为0.15~1.15 mg·ml^(-1)(r=0.9999,n=5),他唑巴坦的线性范围为0.02~0.15 mg·ml^(-1)(r=0.9999,n=5);哌拉西林、他唑巴坦加样平均回收率分别为99.69%(RSD为0.28%)、100.05%(RSD为0.40%)。在柱温(±2℃)、流速(±0.1 ml·min^(-1))、流动相pH值(±0.1)、色谱柱条件做微小变动时,哌拉西林含量最大偏差<1.0%,他唑巴坦含量最大偏差<2.0%。结论本方法简便快速、专属性强、耐用性好,可用于注射用哌拉西林钠/他唑巴坦钠的含量测定和质量控制。

Objective To establish a high performance liquid chromatography(HPLC)method for simultaneous determination of two components in piperacillin sodium and tazobactam sodium in the compound preparation injection.Methods The drugs were analyzed on a reverse phase HPLC system with a C18 column by gradient elution.Mobile phase A consisted of methanol while mobile phase B was composed of water,0.2 mol·L^(-1) sodium dihydrogen phosphate solution and 0.4 mol·L^(-1) tetrabutylammonium hydroxide solution(853∶143∶4).The detection wavelength was 220 nm.The flow rate was 1.0 ml·min^(-1).The injection volume was 10μl.Results The calibration curves of piperacillin and tazobactam were linear in the range of 0.15 to 1.15 mg·ml^(-1)(r=0.9999),and 0.02 to 0.15 mg·ml^(-1)(r=0.9999),respectively.The average recovery of piperacillin and tazobactam was 99.69%(RSD=0.28%)and 100.05%(RSD=0.40%),respectively.When there were slight changes in such conditions as the column temperature(±2℃),flow rate(±0.1 ml·min^(-1)),mobile phase pH value(±0.1)and chromatographic column,the maximum deviation of piperacillin and tazobactam content was less than 1.0%and 2.0%respectively.Conclusion This method is simple,rapid,specific and durable,which can be used for content determination and quality control of piperacillin sodium/tazobactam sodium for injection.