DEVELOPMENT AND VALIDATION OF A HPLC-UV METHOD FOR SIMULTANEOUS DETERMINATION OF PHENYLEPHRINE HYDROCHLORIDE AND KETOROLAC TROMETHAMINE IN OCULAR FORMULATION

A simple, rapid, and sensitive high-performance liquid chromatographic method with UV detection has been developed and validated according to the ICH guidelines for the quantitation of phenylephrine Hydrochloride (PHH), Ketorolac Tromethamine (KET) in pharmaceutical dosage form. Chromatographic separation were achieved on C18 column (250 mm × 4.6 mm; 5 µm particle size) with simple mobile phase composition of 10 mM Potassium dihydrogen phosphate buffer with Triethylamine (pH 3.14) and acetonitrile (40:60, v/v) at a flow rate of 0.5 mL min-1 where detector was set at 302 nm with a total run time of 8 mins. The method was linear over the concentration range of 40-100, µg mL-1 with a correlation coefficient of 0.9891 and 0.994. Limit of quantifications (LOQ) of 13.3, 26.3 and limit of detections (LOD) 4.4, 8.7 µg mL-1 for PHH, and KET respectively. Accuracy and precision values of both within-run and between-run obtained from six different sets of three quality control (QC) samples analyzed in separate occasions for both the analytes ranged from 98.15% to 99.85% and 0.95% to 2.13%, respectively. Extraction recovery of analytes in pharmaceutical formulation from 97.72% to 99.78%. The developed and validated method was successfully applied to quantitative determination of PHH and KET in pharmaceutical formulation. Keywords: High performance liquid chromatography, UV, Spectrophptometry, Phenylephrinehydrochloride & Ketorolac Tromethamine.