RP-HPLC Method Development and Validation of Stability Indicating Assay for Simultaneous Determination of Pantoprazole, Diclofenac, Chlorzoxazone in Pharmaceutical Dosage Form

Background: High-performance liquid chromatography (HPLC) is the fastest-growing analytical technique for the analysis of drugs. Its simplicity, high specificity and wide range of sensitivity make it ideal for the analysis of many drugs in both dosage forms and biological fluids.

Objective: A simple, accurate, precise method was developed, for the simultaneous estimation of the Chlorzoxazone, Pantoprazole, and Diclofenac in solid dosage form.

Method: The chemicals used were Water-HPLC grade, Acetonitrile, Triethylamine, Potassium dihydrogen orthophosphate, and Orthophosphoric acid. Chromatogram was run through BDS C18150×4.6mm, 5m. Mobile phase containing 0.01 % KH2PO4 and Acetonitrile in the ratio of 52:48 v/v was pumped through column at a flow rate of 1.0ml/min. The buffer used in this method was 0.01% KH2PO4, Temperature was maintained at 30°C. The optimized wavelength for Phar Chlorzoxazone, Pantoprazole, and Diclofenac was 229.0 nm.

Results: In the estimation of the three drugs, four trials were conducted using various solvent systems, columns, and experimental conditions. The retention time of Chlorzoxazone, Pantoprazole, and Diclofenac were found to be 2.229 min, 2.958 min, and 3.568 min. The percent RSD of system precision for Chlorzoxazone, Pantoprazole and Diclofenac were 0.9,0.6 And 0.6, respectively. The percent RSD of the method, Pantoprazole and Diclofenac were found to be 0.6, 1.0 and 0.7 respectively. The percent recovery was obtained as 99.86%, 100.07% and 99.70% for Chlorzoxazone, Pantoprazole, and Diclofenac respectively. LOD values obtained from regression equations of Chlorzoxazone, Pantoprazole and Diclofenac were 0.14 ppm, 0.24 ppm, and 1.83 pm, and LOQ values obtained from regression equations of Chlorzoxazone, Pantoprazole and Diclofenac were 0.42 ppm, 0.72 pm, 5.53 ppm respectively. The regression equation of Chlorzoxazone was y = 8321.9.x + 1397.8 Pantoprazole was y = 9806.1x +6071.7 and of Diclofenac was y = 2575x + 4338.4.

Conclusion: Sensitive detectors have transformed liquid column chromatography into high speed, efficient, accurate and highly resolved methods of separation. Retention times were decreased so that the method developed was simple and economical hence it can be used in regular Quality control tests in industries.