An Overview of Developing and Validating HPLC Methods

Abstract

High-performance liquid chromatography (HPLC) is the most commonly used technique for detecting, separating, and quantifying drugs. Various chromatographic parameters (sample pretreatment, mobile phase, column choice, detector type) are considered in the optimisation of HPLC methods. The objective of this article is mainly to describe the processes of method development, optimisation, and validation. The development and validation of analytical methods are critically important for the discovery, development, and manufacture of pharmaceutical drugs and research with humans and animals. HPLC is one of the leading techniques in separation science where analytes are separated by passing through a column packed with micrometre-sized particles. Pharmaceutical industries are required to develop a comprehensive validation policy in accordance with good manufacturing practices (GMP), which describes the approach to validation. This article is focused on the optimisation of HPLC conditions and gives an overview of the introduction, principles, types, instrumentation, method development, and validation processes in HPLC. Validation of HPLC methods provides important information regarding the range, linearity, specificity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), robustness, ruggedness, and system suitability. These validation processes must adhere to regulatory guidelines, for example, those from the International Council for Harmonisation (ICH.