High Pressure Liquid Chromatography (HPLC) is a form of chromatography in which the liquid mobile phase is forced through a column of solid stationary phase under high pressure. HPLC is a powerful analytical tool used to purify, separate, identify, and quantify each component in a mixture.
An HPLC instrument consists of a mobile phase, a pump, an injector, a separation column, and a detector. The sample material is analyzed by pumping a liquid phase (solvent) mixed with the sample through a column packed with a solid phase material. Each component in the mixture responds differently to the absorbent material and causing the constituents of the sample material to separate and come out of the column at different times. Out comes first the material that binds well to the liquid phase and secondly, the material that binds well to the solid phase. HPLC is used to measure the content of various constituents of sample material or for purification. As an example, it can be used in the pharma industry, where HPLC is very useful in measuring the level of active drug in a given material.