高效液相色谱法同时测定小鼠血浆中的利福平与舒巴坦

Simultaneous determination of rifampicin and sulbactam in mouse plasma by high-performance liquid chromatography.

作者信息

Aparicio Irene, Bello Miguel Angel, Callejón Manuel, Guiraúm Alfonso

机构信息

Department of Analytical Chemistry, University of Seville, 41011 Seville, Spain.

出版信息

Biomed Chromatogr. 2006 Aug;20(8):748-52. doi: 10.1002/bmc.591.

DOI:10.1002/bmc.591

PMID:16252264

Abstract

A simple and rapid high-performance liquid chromatographic (HPLC) method with ultraviolet detection has been developed and validated for the simultaneous determination of rifampicin and sulbactam in mouse plasma. Plasma samples were deproteinized with acetonitrile and separated by HPLC on a RP-18 (125 x 4 mm, 5 microm) column and gradient elution with potassium dihydrogen phosphate solution (pH 4.5; 50 mm) and acetonitrile at a flow-rate of 1.0 mL/min. Rifampicin and sulbactam were monitored at 230 nm and confirmed by means of their UV spectra using a diode-array detector. The method was linear at plasma levels from 1 to 100 microg/mL for rifampicin and from 5 to 200 microg/mL for sulbactam. The limits of quantification were 0.6 microg/mL for rifampicin and 4.2 microg/mL for sulbactam. The intra- and inter-day precisions of the method (RSD) were lower than 5% for both compounds. Average recoveries of rifampicin and sulbactam from mice plasma were 98.2 and 89.3%, respectively. The developed method was successfully applied to the determination of the pharmacokinetic profile of both compounds in mice.

摘要

已开发并验证了一种简单快速的带紫外检测的高效液相色谱（HPLC）方法，用于同时测定小鼠血浆中的利福平与舒巴坦。血浆样品用乙腈进行脱蛋白处理，然后在RP - 18（125×4 mm，5微米）柱上通过HPLC进行分离，采用磷酸二氢钾溶液（pH 4.5；50 mM）和乙腈进行梯度洗脱，流速为1.0 mL/min。利福平和舒巴坦在230 nm处进行监测，并使用二极管阵列检测器通过其紫外光谱进行确证。该方法在利福平血浆浓度为1至100 μg/mL以及舒巴坦血浆浓度为5至200 μg/mL范围内呈线性。利福平的定量限为0.6 μg/mL，舒巴坦的定量限为4.2 μg/mL。两种化合物的日内和日间精密度（RSD）均低于5%。利福平和舒巴坦从小鼠血浆中的平均回收率分别为98.2%和89.3%。所开发的方法成功应用于测定两种化合物在小鼠体内的药代动力学特征。