Vol. 4, Issue 7 (2015)

Author(s):
Brijesh Joshi, Prasanna Ku Pradhan, UM Upadhyay, Pratik Pajput

Abstract:
A simple, rapid, accurate, precise and inexpensive reverse phase- High performance Liquid Chromatography (RP-HPLC) method was developed for simultaneous estimation of Naphazoline HCl (NAPH) and Phenylephrine HCl (PHE) utilizing isocratic separation mode with Grace C18, (250 mm X 4.6 mm i.d., 5 μm particle size) column. Optimum mobile phase consisted of Potassium Dihydrogen Phosphate buffer pH 3: Acetonitrile: Triethyleamine in the ratio of 80:20:0.01 v/v/v (pH 3 adjusted with o-Phosphoric acid) with effluent flow rate of 1 ml/min and detection wavelength at 276 nm. The column temperature was kept ambient. Naphazoline HCl and Phenylephrine HCl were found at retention time of 5.976 and 3.154 minutes respectively. Linearity was established in the range of 10-60μg/ml for NAPH and 24-144μg/ml for PHE. With correlation coefficient 0.9997 and 0.9998 respectively. The LOD and LOQ values were found to be 0.28 and 0.85μg/ml for NAPH and 0.48 and 1.47μg/ml for PHE respectively. The developed method can be used for routine quality control analysis of both drugs in eye drops formulation.