HPLC法测定注射用哌拉西林钠舒巴坦钠有关物质

HPLC method used in determining related substances of piperacillin sodium and sulbactam sodium for injection

目的建立注射用哌拉西林钠舒巴坦钠有关物质测定方法。方法采用C18柱；流动相为：水、乙腈、0．2mol／L磷酸二氢钾溶液，线性梯度洗脱；检测波长为220nm。结果线性范围为舒巴坦1．5～36．3μg／mL，1-乙基哌嗪-2，3-二酮1．7～40．6μg／mL，氨苄西林1．4～34．5μg／mL，哌拉西林1．6～38．2μg／mL；重复性为1-乙基哌嗪-2，3-二酮RSD4．8％，氨苄西林RSD14．1％，杂质ⅡRSD4．1％；1-乙基哌嗪-2，3-二酮的回收率均值为（98．9±7．8）％，氨苄西林回收率均值为（103．8±9．7）％；哌拉西林的定量限为4ng，检测限为1ng。样品溶液不稳定，需要配制后立即进样，耐用性试验显示需要选定专用色谱柱。结论本方法简便，专属性、重复性良好，可用于注射用哌拉西林钠舒巴坦钠有关物质的测定。

Objective To establish a method for the determination of related substances of piperacillin sodium and sulbactam sodium for injection. Methods The C18 column was used; The moving phase included water, acetonitrile and 0.2mol/L monopotassium solution, and the linear gradient elution was used; The detection wavelength was 220nm. Results The linear range was 1.5-36.3 μ g/mL for sulbactam, 1.7-40.6 μg/mL for 1-ethyl-piperazine-2, 3-dione, 1.4-34.5 μg/mL for ampicillin, 1.6-38.2 μg/mL for piperacillin respectively; The repeatability was RSD 4.8% for 1-ethyl-piperazine-2, 3-dione, RSD 14.1% for ampieillin and RSD 4.1% for impurity II respectively; The recovery rate was （98.9 ±7.8）% for 1-ethyl-piperazine-2, 3-dione and （103.8 ± 9.7）% for ampieillin respectively; The quantitative limit of piperacillin was 4ng and the detection limit was lng. The sample solution was unstable and needed immediate sample introduction after preparation. Durability tests showed the need for dedicated chromatographic column. Conclusion This method is simple and convenient and has good specificity and repeatability, thus can be used for the determination of related substances of piperacillin sodium and sulbactam sodium for injection.