Method Development and Validation for the Simultaneous Estimation of Ascorbic acid, Phenylephrine HCl, Paracetamol and Levocetirizine HCl using RP-HPLC

An accurate, precise RP-HPLC method was developed for the simultaneous estimation of Ascorbic acid (ASC), Phenylephrine HCl (PHE), Paracetamol (PAR), Levocetirizine HCl (LEV) in tablet dosage form and has been validated as per ICH guidelines. A gradient method was developed using column Aegispak C18 (25cm x 4.6mm, 5μm) and phosphate buffer pH 4.0±0.05, Acetonitrile. The detection was done using wavelength of 220nm. The flow rate was 1.5mL/minute and load volume was 50μL. The retention time of ascorbic acid, phenylephrine HCl, paracetamol, levocetirizine HCl was found to be 2.4±0.1, 5.4±0.1, 10.0±0.1, and 22.7±0.1 respectively. The method was linear in concentration ranges of 50 to 150% and R2 was found to be with in the limit. The result of accuracy was found to be between 98 to 102%. The method was precise as the % RSD was found that NMT 2.0%. Specificity studies reveals no interference in the standard peaks due to placebo and diluent effect. The developed precise method was validated in respect to linearity, range, precision, accuracy, robustness.