RP-HPLC-UV Validation Method for Levofloxacin Hemihydrate Estimation in the Nano Polymeric Ocular Preparation

The specific and accurate reversed-phase HPLC-UV method has been validated to determine levofloxacin hemihydrate (LEVH) level. The separation was conducted at C 18 analytical column by administering mobile phase acetonitrile, methanol, and phosphate buffer (pH 3) with the ratio of 17:3:80. The flow rate of the mobile phase was 1 mL/min with a UV detector at 295 nm wavelength. Analytical methods validation evaluated includes specificity, linearity, accuracy, precision, LOD, LOQ, and robustness. The implementation of the analytical method was employed to determine LEVH level in ocular polymeric nanoparticles preparations. The test was specific for LEVH with the retention time of 7.66 min. Linearity was obtained from the concentration range of 4.8 µg/mL to 29.04 µg/mL. All method validation criteria are within the acceptable range. The developed method can be applied for LEVH polymeric nano-formulation analysis.