Reversed Phase High Performance Liquid Chromatography Method Development and Validation for Simultaneous Estimation of Dicyclomine Hydrochloride, Paracetamol and Mefenamic Acid in Bulk and Tablet Dosage Form

Abstract

Objective: The objective of this study was to develop and validate a simple, precise and accurate reverse phase high performance liquid chromatographic method for simultaneous estimation of dicyclomine hydrochloride, paracetamol, and mefenamic acid from multicomponent tablet dosage form. Methods:The optimized chromatographic separation was achieved using a stationary phase of hypersil gold ODS-C18 (250 × 4.6 mm i.d, 5 μm particle size) column and mobile phase of acetonitrile: Methanol: Potassium dihydrogen phosphate buffer, pH adjusted to 4.5 with orthophosphoric acid (OPA) in the ratio of 50: 30: 20 (v/v/v) with a flow rate of 1 mL/minute. The ultraviolet detection was carried out at 260 nm. Results:The retention times of paracetamol, dicyclomine hydrochloride and MEF were found to be 4.2, 6.1 and 3.4 minutes, respectively. The method was validated for linearity, precision, accuracy, and robustness. The developed method provided linear responses within the concentration ranges 2-12 µg/mL for dicyclomine hydrochloride, 50-300 µg/mL for paracetamol, and 25-150 µg/mL for MEF. The % recovery obtained were found to be 99.09-99.44% for dicyclomine hydrochloride, 99.22-99.51% for paracetamol, and 99.49-99.60 for MEF. Conclusion:The proposed method enables rapid quantification and simultaneous analysis of all the drugs from commercial formulations without any interference of excipients. Hence, the developed method can be successfully applied for routine analysis of dicyclomine hydrochloride, paracetamol, and MEF in their combined tablet formulation.