Abstract

HIGH- PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) METHOD DEVELOPMENT AND VALIDATION

Priyanka Chirde*, A. P. Dewani and Dr. A. V. Chandewar

ABSTRACT

High Performance Liquid Chromatography (HPLC) is a type of column chromatography which is used in Analytical chemistry and also in Biochemistry HPLC is the dominant separation technique to detect, separate and quantify the active components. HPLC method depends on chemical structure of molecules, synthetic route, solubility, polarity, pH, pKa value and functional activity group. Chromatography is a technique which is used for separation of constituents in a mixture. This involves two phases Stationary phase and Mobile phase Stationary phase used is column and mobile phase is solvent used for separation. There are different types of chromatography i.e Paper Chromatography, Gas chromatography, Thin Layer Chromatography, Ion exchange Chromatography, High Performance Thin Layer Chromatography, Reverse-Phase High Performance Liquid Chromatography In HPLC method the solvent will be forced under pressure 400 atmosphere so that the sample can be separated. Technique documentation is very important for activity in pharmaceutical evaluation to ensure that analytical methods are suitable for intended use. The objective of this article is to review the method development, optimization and validation Validation of HPLC method as per ICH Q2 (R1) Guideline gives information regarding various stages and knowing characteristics like system suitability, Accuracy, specificity, linearity, precision, robustness, limit of detection, limit of quantification.

Keywords: High-Performance Liquid Chromatography (HPLC), Method validation, Method development.