Analytical Method Development, Optimization and Validation of Combination of Azithromycin and Benzoyl Peroxide By RP-HPLC Using Design of Experiment as Per ICH Guideline

Abstract

A simple, fast, accurate and precise RP-HPLC method were developed and validated for the estimation of Azithromycin & Benzoyl peroxide per ICH guidelines. Potassium dihydrogen phosphate and Acetonitrile (50:50) are commonly used as solvents. The method was developed in Eclipse C18 column (Waters XTerra®, 4.6Χ250 mm, particle 5μ) with Potassium dihydrogen phosphate and Acetonitrile are commonly used solvents in RP -HPLC having low UV cut- off of 200-400 nm respectively. In RP-HPLC method was found to be linear in the range of Azithromycin/Benzoyl peroxide is 1-5μg/ml with a correlation coefficient value of 0.99. The accuracy studies of RP-HPLC method was performed at three different levels, i.e., 80%, 100%. The limit of detection (LOD) and Limit of Quantification (LOQ) for Azithromycin 0.009μg/ml & 0.028μg/ml and for Benzoyl peroxide 0.015μg/ml & 0.046μg/ml is to be were found. The Percentage RSD should not be more than 2 which indicate the accuracy and precision of the method. Hence the method was robust.Thus this shows that the method is capable to give a good detector response, the recovery calculated was within the range of 98% to 101% of the specification limits. Hence the method was a rapid tool for routine analysis of Azithromycin/Benzoyl peroxide in the bulk and in the pharmaceutical dosage form.