Abstract

A simple, fast, new, precise, sensitive, and accurate reverse-phase high performance liquid chromatography method was developed and validated according to International Council for Harmonisation guidelines for the estimation of enrofloxacin and ketoprofen in bulk and marketed formulation. Chromatographic separation was achieved using a Shimadzu high performance liquid chromatography system with an inertsil octadecylsilyl C₁₈ column (4.6 mm×250 mm internal diameter, 5 µm particle size). The best results were obtained with the mobile phase composition consisting of 0.1 % trifluoroacetic acid, methanol, and acetonitrile in a ratio of 20:40:40 v/v/v. The system was regulated at a 0.7 ml/min flow rate at an optimized wavelength selected for detection at 262 nm. The retention time for enrofloxacin and ketoprofen were 2.941 and 5.756 min, respectively. The method has been validated for linearity, accuracy, precision, limit of detection, limit of quantification, and robustness as per International Council for Harmonisation guidelines. The calibration graphs were linear over the concentration range of 10-50 µg/ml for enrofloxacin and 6-30 µg/ml for ketoprofen. The values for enrofloxacin and ketoprofen were 1 µg/ml and 0.6 µg/ml for the limit of detection, and 0.3 µg/ml and 1.8 µg/ml for the limit of quantification, respectively. The analysis's conclusion indicates that for all of the validation parameters, the % relative standard deviation will be less than 2, and recovery studies revealed that the results were within the predetermined bounds. Therefore, it was determined that the suggested method was effective and that it could be utilized for the routine examination of enrofloxacin and ketoprofen in their marketed formulation.