Simultaneous determination of cefdinir and cefixime in human plasma by RP-HPLC/UV detection method: Method development, optimization, validation, and its application to a pharmacokinetic study

摘要

A novel isocratic reversed-phase high performance liquid-chromatography/ultraviolet detection method for simultaneous determination of cefdinir and cefixime in human plasma was developed and validated after optimization of various chromatographic conditions and other experimental parameters. Sample preparation based on a simple extraction procedure consisting of deproteination and extraction with 3 parts of 6% trichloroacetic acid aqueous solution followed by volume make up with the aqueous component of the mobile phase obtained best recoveries of the two analytes. Samples were separated on a Supelco Discovery HS C_(18) (150mm×4.6mm, 5μm) analytical column protected by a Perkin Elmer C_(18) (30mm×4.6mm, 10μm) guard cartridge. The mobile phase, methanol/acetonitrile (50/50, v/v):0.05% trifluoroacetic acid (19:81, v/v), operated at 50°C column oven temperature was pumped at a flow rate of 2.0mLmin~(-1) and the column eluents were monitored at a wavelength of 285nm. When Sample was injected into the Perkin Elmer high performance liquid-chromatography system through Rheodyne manual (or auto-sampler) injector equipped with 20μL loop, separation was achieved within 4min. The present method demonstrated acceptable values for selectivity, linearity within the expected concentration range (0.004-5.0μgmL~(-1); r~2 >0.999 for both analytes), recovery (>95% for cefdinir and >96% for cefixime), precision (%RSD <2.0 for cefdinir and <2.2 for cefixime), sensitivity (limit of detection: 1ngmL~(-1) and lower limit of quantification: 4ngmL-1 for both analytes), stability of solutions, and robustness. The method was efficiently applied to a pharmacokinetic study in healthy volunteers.