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Column chromatography in chemistry is a chromatography method used to isolate a single chemical compound from a mixture. Chromatography is able to separate substances based on differential absorption of compounds to the adsorbent; compounds move through the column at different rates, allowing them to be separated into fractions.
Chromatography columns of different types are used in both gas and liquid chromatography: Liquid chromatography: Traditional chromatography columns were made of glass. Modern columns are mostly made of borosilicate glass, acrylic glass or stainless steel. To prevent the stationary phase from leaking out of the column interior a polymer ...
Liquid chromatography as we know it today really got its start in 1969, when the first modern HPLC was designed and marketed as a nucleic acid analyzer. [9] Columns throughout the 1970s were unreliable, pump flow rates were inconsistent, and many biologically active compounds escaped detection by UV and fluorescence detectors. Focus on ...
The mobile phase moves through the chromatography column (the stationary phase) where the sample interacts with the stationary phase and is separated. Preparative chromatography – the use of chromatography to purify sufficient quantities of a substance for further use, rather than analysis.
High performance affinity chromatography (HPAC) [33] works by passing a sample solution through a column packed with a stationary phase that contains an immobilized biologically active ligand. The ligand is in fact a substrate that has a specific binding affinity for the target molecule in the sample solution.
For example, one class of the most commonly used chiral stationary phases both in liquid chromatography and supercritical fluid chromatography is based on oligosaccharides [4] such as Amylose Cellulose or Cyclodextrin (in particular with β-cyclodextrin, a seven sugar ring molecule) immobilized on silica gel.
Typically, when an aqueous solution is used to transport the sample through the column, the technique is known as gel-filtration chromatography, versus the name gel permeation chromatography, which is used when an organic solvent is used as a mobile phase.
[13] [23] A compound elutes from a column when the amount of solvent collected is equal to 1/R f. [24] The eluent from flash column chromatography gets collected across several containers (for example, test tubes) called fractions. TLC helps show which fractions contain impurities and which contain pure compound.