A Validated RP-HPLC Method for Simultaneous Assay Determination and UV-Spectrophotometric Method for Dissolution Estimation in Film-Coated Tablets Containing Cefixime and Clavulanic Acid

Abstract

A simple and novel reverse phase high performance liquid chromatography (RP-HPLC) method was developed for the simultaneous assay estimation of film coated tablets incorporating clavulanic acid (CVA) and cefixime (CFX). Furthermore, the same method was also applied in order to assess the dissolution of CVA while a UV spectroscopic method was produced to examine the drug release of CFX. In this research, an analytical column used was C18 (4.6-mm × 25-cm); 5-μm. Moreover, the mobile phase was prepared by completely dissolving 3.408 gm of disodium hydrogen phosphate in 800 mL of water (HPLC grade), 200 mL methanol was added in (20:80 v/v) it and then finally pH was adjusted to 6.0 using orthophosphoric acid. In addition, flow rate, detecting wavelength, and run time were 1.0 mL/min, 220 nm, and 15 minutes, respectively. Furthermore, the maximum absorption of CFX was obtained at 288 nm thereby this wavelength was optimized for the dissolution estimation of CFX by UV spectroscopy. The validation parameters were validated according to the specifications and guidelines of the international conference on harmonization (ICH) prior to evaluating both assay and dissolution of CVA and CFX. In case of assay estimation, the findings of the linearity revealed a linear interrelation of concentration between 0.16 - 0.25 mg/mL for both CVA and CFX. Similarly, the calibration curve was linear between the concentration range of 0.03 – 0.045 mg/mL and 0.004 – 0.006 mg/mL, respectively for dissolution determination. Additionally, the value of LOD and LOQ of CVA for assay evaluation was obtained 0.011 and 0.033 μg, respectively. The accuracy of the developed method was assessed by determining recovery studies and the mean recovery of CVA and CFX obtained 99.35% and 100.50%, respectively at 100% spiked level for assay analysis. The repeatability evaluation for assay and dissolution demonstrated that the optimized method is precise within the appropriate range and the RSD% was observed lower than 2.0%. In addition, results of different validation parameters including study of solution stability were achieved within the suitable limit as well.