The development and validation of a reversed‐phase liquid chromatographic method for the determination of lomefloxacin in seminal plasma is described. Lomefloxacin was extracted on a solid‐phase cartridge and separated on a reversed‐phase column with acetonitrile in phosphate buffer at pH 7.0 as mobile phase. The solid‐phase extraction showed high recovery for lomefloxacin from seminal plasma samples. The chromatographic peaks height ratio of lomefloxacin and internal standard, obtained by fluorimetric detection, was used for quantitative analysis. The proposed method was validated with respect to accuracy, precision, linearity, and specificity. Also, the method was determined to be robust with regards to the following parameters: mobile phase, apparent pH; mobile phase organic content. The percent recoveries of lomefloxacin, the limit of detection (LOD) and limit of quantitation (LOQ) for the HPLC method have been determined. This high‐performance liquid chromatographic method has been successfully used in medical laboratories to assay seminal plasma samples for studies on the treatment of chronic bacterial infections with lomefloxacin.
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