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    Description of the Chromatography Technique

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    What is Chromatography?
    How do differences in polarity and molecular weight between amino acids affect migration in a mixture of proteins?

    © BrainMass Inc. brainmass.com October 4, 2022, 1:00 pm ad1c9bdddf

    SOLUTION This solution is FREE courtesy of BrainMass!

    Chromatography is a technique that is used to separate a mixture of substances. The mixture that is to be separated is dissolved in a liquid called the mobile phase. The mobile phase is transported (usually by capillary action) through another material called the stationary phase. The separation of the constituents of the mobile phase is based on a number of factors.

    The separation of a mixture of amino acids in the mobile phase depends on the polarity and molecular weight of the individual amino acids.

    The amino acids will solubilize (dissolve) in the mobile phase as it passes through the stationary phase. Amino acids at the molecular level have a structure that can have different polarities. These differences in polarity arise from the different molecular structure of each amino acid. The affinities of these amino acids for the mobile phase can be correlated to the solubility of the different amino acids in the solvent (i.e., an amino acid that is highly soluble in the eluting solvent will have a higher affinity for the mobile phase than an amino acid that is less soluble in the solvent.). So, amino acids that have a greater affinity (greater solubility) to mobile phase will therefore travel farther through the stationary phase. The end result is that the individual amino acids will separate out according to their polarity (i.e. affinity/solubility).

    Amino acids all have different molecular weights. As the amino acids are transported through the stationary phase, the smaller, lighter amino acids will travel faster than the heavier bulkier amino acids.

    The combination of polarity and molecular weight allows you to separate a mixture of amino acids into individual amino acids

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    1. Mathew DC, Luthey-Schulten Z. On the physical basis of the amino acid polar requirement. J Mol Evol. 2008 May;66(5):519-28.
    2. Still, W. C.; Kahn, M.; Mitra. A. Rapid chromatographic technique for preparative separations with moderate resolution. J. Org. Chem. 1978, 43(14), 2923?2925.

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