Development and validation of HPLC method - a review

Abstract

Many different strategies of high performance liquid chromatographic method development are used today. This review describes a strategy for the systematic development of High performance liquid chromatographic (HPLC) methods. HPLC is an analytical tool which is able to detect, separate and quantify the drug, its various impurities and drug related degradants that can form on synthesis or storage. It involves the understanding of chemistry of drug substance and facilitates the development of analytical method. A number of chromatographic parameters were evaluated in order to optimize the method. An appropriate mobile phase, column, column temperature, wavelength and gradient must be found that affords suitable compatibility and stability of drug as well as degradants and impurities. Forced degradation or alternatively referred as stress testing and it demonstrates specificity when developing stability indicating methods, especially when little is known about potential degradation products. Force degradation studies are helpful in development and validation of stability-indicating methodology, determination of degradation pathways of drug substances and drug products, discernment of degradation products in formulations that are related to drug substances versus those that are related to non–drug substances (e.g. excipients).