Journal of Biochemistry Research

High Performance Liquid Chromatography Scholarly Open Access Journals

High Performance Liquid Chromatography ( HPLC) is a form of column chromatography that pumps a sample mixture or analyte through a column with chromatographic packaging material (stationary phase) at high pressure in a solvent (known as the mobile phase). The sample is carried by a flux of moving helium or nitrogen carrier gas. HPLC has the ability to separate and identify compounds present in any sample that may dissolve at trace concentrations as low as parts per trillion in a liquid. HPLC is used in a variety of industrial and scientific applications, such as pharmaceutical, environmental, forensics, and chemicals, because of this versatility. Depending on the interaction between the stationary phase, the molecules being analyzed, and the solvent or solvents used, sample retention time will vary. As the sample passes through the column it interacts at different rates between the two phases, mainly due to different polarities in the analytes.  Analytes that have the least amount of stationary phase interaction or the most amount of mobile phase interaction will exit the column more quickly. The solvent [called the mobile phase, because it moves] is held in a reservoir. Using a high pressure pump, a specified mobile phase flow rate, typically milliliters per minute, is generated and metered. An injector can insert the sample into the continuously flowing mobile phase stream, which carries the sample into the HPLC column. The column contains the material for chromatographic packing required to effect the separation. This packaging material is called the stationary phase, because the column hardware holds it in place. To see the separate compound bands as they elute from the HPLC column, a detector is needed. 

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