Simultaneous Determination of Orphenadrine Citrate and Paracetamol in Tablets by using RP- HPLC Coupled with UV Detection

Abstract

A simple, sensitive, specific, and cost effective method for simultaneous determination of Orphenadrine Citrate and Paracetamol was developed and validated in single dosage formulation. The sample solution of PA and ORC was prepared using methanol as a solvent. Separation of PA and ORC was achieved with a mobile phase consisting of 1% Triethylamine aqueous: Methanol: Acetonitrile (35:20:45 v/v) at a flow rate of 2.0 ml/min and the wavelength of 220nm. Separations were performed on Merck Hibar 250-4.6 RP18 (5 µm) column (250 mm X 3.4 mm), using a Lab Alliance HPLC system equipped with a UV-Vis detector, manual injector with 20 μL loop, LC pump, Clarity controller and software. Retention times of PA and ORC were 2.265 and 3.882 minutes respectively. Absolute recovery of PA and ORC was 100.20 and 100.07 % respectively. The lower limit of quantification (LLOQ) of PA and ORC was 0.3097 and 0.1063 ppm and lower limit of detection (LLOD) of PA and ORC was 0.0153 and 0.0135 ppm respectively. Linearity was established for the range of concentrations (20-140) μg/ml and (0.1-50) μg/ml for PA and ORC respectively with the coefficient of determination (R²) of 0.9991 and 0.9997 for both the compounds. The inter- and intra-day precision in the measurement of PA quality control (QC) sample 450 μg/ml, were in the range 0.1-0.2 % relative standard deviation (R.S.D) and 0.2-0.3 % (R.S.D)., respectively. The inter- and intra-day precision in the measurement of ORC quality control (QC) sample 35 μg/ml, were in the range 0.1-0.2 % (R.S.D)., and 0.0-0.3 % (R.S.D)., respectively. The developed method would be applicable for routine quality control of PA and ORC in bulk as well as in pharmaceutical formulations.

Keywords: Orphenadrine Citrate, Paracetamol, RP-HPLC, Tablets, Validation.