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A Review on Method Development, Validation, Optimization and Applications of HPLC

Abstract: High-Performance Liquid Chromatography (HPLC) is a technique used to separate components from the mixture. The technology consists of two phases, the stationary phase, and the mobile phase. Component separation is based on differences in partition coefficients in two phases. The present review article focuses on the principle, types, instrumentation, process validation, application, advantages, disadvantages of HPLC. HPLC methods development and validation play important roles in the discovery, development, manufacture of pharmaceutical drugs, and various other studies related to humans and animals. An analytical procedure is developed to test a defining characteristic of the drug substance or drug product against established acceptance criteria for that characteristic. Validation of HPLC method as per ICH Guidelines covers all the performance characteristics of validation, like Accuracy, precision, specificity, linearity, range, the limit of detection, the limit of quantification, robustness, and system suitability testing. Keywords: HPLC, Types, Method validation, Method development, Instrumentation, and Applications.