HPLC法测定人脑脊液中美罗培南浓度

Determination of meropenem in human cerebrospinal fluid by HPLC

目的建立美罗培南在人脑脊液中浓度测定的HPLC法。方法色谱柱为C_(18)柱。流动相为乙酸胺:乙腈(90:10,V/V),紫外检测波长308nm。样品加入内标(PABA)后,乙腈直接沉淀,上清液吹干重组,进样量为20μL。结果脑脊液样品中美罗培南保留时间为3.810 min,在脑脊液药浓度0.25～50 mg·L^(-1)内线性关系良好(r=0.999 0);各浓度质控提取回收率在90%～110%之间,日内、日间RSD均<5%;-80℃冰箱保存6个月内回收率稳定。结论本研究建立的人脑脊液中美罗培南检测方法灵敏高、特异性强,适用于临床药动学的研究。

AIM To establish a HPLC method for determination of meropenem in hunman cerebrospinal fluid. METHODS Chromatorgraphic separation was performed on a TSK - gel ODS - 80TM C18 column.The mobile phase was ammonium acetate and acetonetrile（90：10,V/V）.The flow rate was 1.0 mL·min^（-1）.The ultraviolet absorption was set at 308 nm for detection.The temperature of column was 35℃.After protein was precipitated by acetonetrile,the derivatized sample was eluted and 20μL was injected into the column.Para-aminobenzoic acid（PABA） was used as an internal standard.RESULTS In human cerebrospinal fluid,the retention time of meropenem was 3.810 min and the detector response was linear within the concentration range of 0.25 - 50 mg·L^-1.The recovery was between 90%- 110%,and relate standard deviation（RSD） of intra-day and inter-day assays were all less than 5%.Stored at - 80℃for 6 months at various concentrations in cerebrospinal fluid,meropenem did not reveal any appreciable degradation.CONCLUSION The method is simple,sensitive and accurate,and can be used for the pharmacokinetic study.