A very accurate, precise, and reliable isocratic HPLC technique has been developed and verified for the simultaneous quantification of Rifampicin and Ofloxacin. The chromatographic separation was performed using a Phenomenex C18 column with a mobile phase consisting of a combination of 0.05M KH2PO4 buffer at pH 4.0 and methanol in a ratio of 55:35. The separation was monitored at a wavelength of 230 nm. Ofloxacin had a retention time of 4.920 min, whereas Rifampicin had a retention time of 11.710 min. The calibration plots for Rifampicin and Ofloxacin were linear between the concentration ranges of 2–10 µg/ml and 4–20 µg/ml, respectively. The correlation coefficient for Rifampicin was 0.9999, while for Ofloxacin it was 0.9996. The Rifampicin had a limit of detection (LOD) of 0.2345 µg/ml, whereas the Ofloxacin had a LOD of 0.4625 µg/ml.The approach underwent validation to assess its linearity, accuracy, precision, and robustness. The percentage of recoveries was determined to be about 100% with little fluctuation. This approach can be used for regular analysis