Applications of Six Sigma and Quality by Design Approach in UHPLC Method Development for Simultaneous Assessment of Eight Bronchodilators in Drug Substance

Abstract

This work portrays the development and validation of a simple and fast UHPLC method for the simultaneous estimation of eight bronchodilators in the drug substance. Process and performance capability (Cpk, Ppk, PPM) studies were conducted by which it may easily conclude how well developed method meeting the requirements. Integration of quality by design (QbD) concept to analytical method development was also discussed. The temperature and mobile phase interaction on chromatographic separation were studied using design of experiments (DoE). Using the QbD and design of experiments (DoE) as combination, a design space was established for developed method. The developed method facilitates the quantitative determination of clenbuterol hydrochloride, fluticasone, formoterol, glycopyrrolate, levalbuterol, metaproterenol, salmeterol and theophylline as a drug substance. All the compounds were separated within 20 min using analytical column X-Bridge BEH C18; (100 × 2.1 mm, 2.5 μ) mobile phase consisted variable mixtures of mobile phase A (10 mM ammonium acetate pH 4.5) and mobile phase B (methanol) with the gradient elution, column temperature 30 ºC, mobile phase flow rate 0.2 mL min/min, wavelength of detection 234 nm.