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Column chromatography

Column chromatography in chemistry is a chromatography method used to purify individual chemical compounds from mixtures of compounds. It is often used for preparative applications on scales from micrograms up to kilograms. The main advantage of column chromatography is the relatively low cost and disposability of the stationary phase used in the process. The latter prevents cross-contamination and stationary phase degradation due to recycling.

Column chromatography is an extremely time consuming stage in any lab and can quickly become the bottleneck for any process lab. Therefore, several manufacturers like Biotage, Buchi, Interchim and Teledyne Isco have developed automated flash chromatography systems (typically referred to as LPLC, low pressure liquid chromatography, around 350–525 kPa or 50.8–76.1 psi) that minimize human involvement in the purification process. Automated systems will include components normally found on more expensive high performance liquid chromatography (HPLC) systems such as a gradient pump, sample injection ports, a UV detector and a fraction collector to collect the eluent. Typically these automated systems can separate samples from a few milligrams up to an industrial many kilogram scale and offer a much cheaper and quicker solution to doing multiple injections on prep-HPLC systems.

The resolution (or the ability to separate a mixture) on an LPLC system will always be lower compared to HPLC, as the packing material in an HPLC column can be much smaller, typically only 5 micrometre thus increasing stationary phase surface area, increasing surface interactions and giving better separation. However, the use of this small packing media causes the high back pressure and is why it is termed high pressure liquid chromatography. The LPLC columns are typically packed with silica of around 50 micrometres, thus reducing back pressure and resolution, but it also removes the need for expensive high pressure pumps. Manufacturers are now starting to move into higher pressure flash chromatography systems and have termed these as medium pressure liquid chromatography (MPLC) systems which operate above 1 MPa (150 psi).

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