METHOD DEVELOPMENT AND VALIDATION OF ANTIBACTERIAL DRUGS BY USING RP-HPLC METHOD IN SOLID DOSAGE FORM

Abstract

The analytical method was developed by studying different parameters. First of all, maximum absorbance was found to be at 230nm and the peak purity was excellent. Injection volume was selected to be 10µl which gave a good peak area. The column used for study was WATERS RP-C18 (250mm×4.6mm, 5µm) produce good peak shape. Ambient temperature was found to be suitable for the nature of drug solution. The flow rate was fixed at 1.0ml/min because of good peak area and satisfactory retention time. Different ratios of mobile phase were studied, mobile phase with ratio of 70:30v/v orthophosphoric acid Buffer: Methanol was fixed due to good symmetrical peak. So this mobile phase was used for the proposed study. Methanol was selected because of maximum extraction sonocation time was fixed to be 15min at which all the drug particles were completely soluble and showed good recovery. Run time was selected to be 6min because analyte gave peak around 3.09 and 4.55 for Levofloxacin and Cefixime respectively and also to reduce the total run time. This system produced symmetric peak shape, good resolution and reasonable retention times of Cefixime and Levofloxacin. The mean recoveries were found to be 100 and 100 respectively for Levofloxacin and Cefixime respectively was linear and precise over the same range. Both system and method precision was found to be accurate and well within range.  Detection limit was found to be 0.006 and 0.002 respectively for Cefixime and Levofloxacin. Linearity study was, correlation coefficient and curve fitting was found to be 0.99 and 0.99 for Cefixime and Levofloxacin respectively. The analytical method was found linearity over the range of 50-150ppm of the target concentration. The average retention time for Cefixime and Levofloxacin were found to be 4.56 and 3.09 respectively.  The analytical passed robustness test. Relative standard deviation was well satisfactory.