HPLC: Principles and Applications

Abstract

HPLC is a chromatographic technique used to separate a mixture of compounds in analytical chemistry and biochemistry with the purpose of identifying, quantifying or purifying the individual components of a mixture. A HPLC system has basically a stationary phase (column) through which the mobile phase carrying the sample is passed and the analytes (compounds to be separated) are eluted from the column depending upon their chemical structure and interaction with the column. Different components of a complex mixture get separated in the column due to various physico-chemical interactions with the column. HPLC has many uses including medical diagnosis (e.g. detecting vitamin D levels in blood serum), legal (e.g. detecting performance enhancement drugs in urine), research (e.g. separating the components of a complex biologicalsample, or of similar synthetic chemicals from each other), and manufacturing (e.g. during the production process of pharmaceutical and biological products) and quality control (quantification of certain compounds in a product).