Until now, analysis of Ofloxacin in human blood plasma using solidphase extraction (SPE) by HPLC UV detector has not been reported. This study aims to determine the validity of analytical methods in Ofloxacin study in human blood plasma (in vitro) using an HPLC SPE UV detector. Plasma samples were extracted by SPE. Analytes were analyzed using a C18 column (octadecylsilane) 250x4.6 mm, particle size 10 0m, mobile phase 85,5:14,5 v v 0.025 M phosphate buffer (pH 2.2) and acetonitrile with a flow rate of 2 ml/min, detection performed at 294 nm with the internal standard ciprofloxacin. Validated analytical method was based on the parameters: selectivity, accuracy, precision, repeatability, linearity, LOD, LOQ, and the suitability of the system. Validation analysis showed selectivity test Rs>1.5, test repeatability with CV(%) <10%, linearity was obtained in the range of 0.1 to 6 ;g/ml with correlation coefficient (r) from 0.9998 to 0.9999. Based on the area ratio of peak height and a segment of the chromatogram obtained LOD values 0.023 and 0.024 ;g/ml, LOQ value of 0.076 and 0.080 tg/ml, percent accuracy from 94.32 to 100.45% and 97.68 to 101.63%, and precision CV (%) 0.31 to 0.85% and 0.84 to 1.08%. System suitability test results on the retention time, area ratio, and high ratios of peak chromatogram shows the CV(%) <10%. Can be concluded that the analytical methods used have validity in accordance with the requirements.