Application of Central Composite Design to the Optimization of HPLC Analysis of Nitroimidazoles

Abstract

A 2 factor central composite design is used to study the effects of acetonitrile concentration and eluent pH on the retention and resolution of metronidazole and tinidazole on a C-18 column. The equations obtained have been used to predict the composition of an eluent for an optimum separation. The assay developed has been applied to the quantitation of metronidazole in human plasma following simple protein precipitation. The detection limit at 320nm for metronidazole is 0.01μg/mL. Using this assay a relative bioavailability study of two commercial metronidazole tablets in ten healthy volunteers has been performed.