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    Gel chromatography used to test Polymer standard

    来源:cxbio.com 作者:女足世界杯开户 人气:- 发表时间:2012-06-21 10:39:13【

    Gel chromatography
    gel-chromatographyGel chromatography separates bimolecular on the basis of size. This technique uses a molecular sieve in the form of polysaccharide gel that comprises spherical beads. As these beads are tiny and porous, so they allow only smallmoleculesto enter the gel, while the largermoleculesremain outside these beads. Therefore, when the column is eluted with an appropriate buffer, largermoleculespassout through the column. At a faster rate then smaller ones. This technique is also called as size exclusive chromatography.

    Phases of chromatography

    The components to be separated are distributed between two phases: astationaryphase and a mobile phase, which percolates through astationarybed. Thestationaryphase basically comprises a column field with porous beads.
    Column packing
    Spherical porous beads of defined sizes are used to pack the chromatography column. The porous gel may comprise cross-linked dextrans (sephadex), cross-linked poly-acrylamide (bio- gel P) or cross-linked agarose (sepharose). When columns are created they are packed with porous beads with a specific pore size so that they are mostaccurateat separatingmoleculeswith similar to the pore size.


    Method

    Asamplecomprising a mixture ofproteinsis dissolved in the same solvent that is running through the column. The variousmoleculesof this mixture get flushed through the column at different rates. The separation of thesemoleculesdepends on whether they can fit with in the pore size of packing material.

    As a molecule flow through the column, itpassesby a no. of porous beads. Smallmoleculesenters the porous beads and hence, their rate of passage through the gel is retarded as compared to that of the larger molecule that can not fit into the pores and continue moving along the solvent. So, molecule with the larger size reaches the end of the column before the molecule with smaller size do. Small amount of solvent eluted through the gel are collected in differenttubesand theproteinpresent in eachsampleare analyzed.

    Applications of the gel chromatography
    Gel chromatography is a very efficient method for the separation, analyzing and characterization of the mixture of macro-molecules.
    1. In nay chemical or bioprocessing industry, separation and purification of a product from complex mixture is very important. Gel chromatography can separate complex mixtures with great precision.
    2. Chromatography is well suited to a variety of uses inthe fieldof biotechnology, such as separating mixture ofproteins.

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