V3I10P27

A Review on High-Performance Liquid Chromatography (HPLC): Principle, Instrumentation, Working, Types and Pharmaceutical Applications

Afruza Begum¹, Kaja Reshma^2*

Abstract

High-performance liquid chromatography (HPLC) is one of the most common analytical methods used in pharmaceutical analysis and quality control. HPLC is a type of liquid chromatography that is used to separate, identify and quantitatively determine the various components present in a mixture. HPLC is characterized by a high level of sensitivity, accuracy, precision, and reproducibility, making it an important technique for the analysis of drugs, formulations, biological samples and other chemical substances. The method involves passing a liquid mobile phase that contains the sample through a column packed with a stationary phase under high pressure. The various components of the sample interact differently with the stationary and mobile phases leading to their separation. Components thus separated are then detected and represented as peaks in a chromatogram. HPLC can be carried out using different chromatographic mechanisms, including adsorption, partition, ion exchange and size exclusion. Reversed phase HPLC is particularly vital in pharmaceutical analysis due to its wide applicability. This article discusses the principle, instrumentation, working procedure, main types, important chromatic parameters, applications, advantages, limitations, and precautions associated with HPLC. The technique is very vital in drug identification, assay, impurity testing, stability studies and quality control of pharmaceutical products.

Keywords:

High-Performance Liquid Chromatography; HPLC; Chromatography; Pharmaceutical Analysis; Separation; Quality Control; Reversed-Phase HPLC