HPLC relies on pumps to pass a pressurized liquid and a sample mixture through a column filled with adsorbent, leading to the separation of the sample components. Ĭhromatography can be described as a mass transfer process involving adsorption. HPLC has been used for manufacturing ( e.g., during the production process of pharmaceutical and biological products), legal ( e.g., detecting performance enhancement drugs in urine), research ( e.g., separating the components of a complex biological sample, or of similar synthetic chemicals from each other), and medical ( e.g., detecting vitamin D levels in blood serum) purposes. Each component in the sample interacts slightly differently with the adsorbent material, causing different flow rates for the different components and leading to the separation of the components as they flow out of the column. It relies on pumps to pass a pressurized liquid solvent containing the sample mixture through a column filled with a solid adsorbent material. High-performance liquid chromatography ( HPLC), formerly referred to as high-pressure liquid chromatography, is a technique in analytical chemistry used to separate, identify, and quantify each component in a mixture. (1) Solvent reservoirs, (2) Solvent degasser, (3) Gradient valve, (4) Mixing vessel for delivery of the mobile phase, (5) High-pressure pump, (6) Switching valve in "inject position", (6') Switching valve in "load position", (7) Sample injection loop, (8) Pre-column (guard column), (9) Analytical column, (10) Detector ( i.e., IR, UV), (11) Data acquisition, (12) Waste or fraction collector.
Schematic representation of an HPLC unit.
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