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    Column Chromatography PDF

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    GLADYS FERNANDES
    Column chromatography is a technique used to separate chemical compounds dissolved in a fluid mixture. It works by differential adsorption of compounds to the stationary phase material in the column, causing the compounds to move through the column at different rates and elute separately. Factors like a compound's affinity and adsorption to the stationary phase determine its rate of movement. This technique can purify materials on both small and large scales for use in experiments. It has applications like isolating active ingredients, separating compound mixtures, drug estimation in formulations, and removing impurities.

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    Column Chromatography PDF

    Uploaded by

    GLADYS FERNANDES
    139 views11 pages
    Column chromatography is a technique used to separate chemical compounds dissolved in a fluid mixture. It works by differential adsorption of compounds to the stationary phase material in the column, causing the compounds to move through the column at different rates and elute separately. Factors like a compound's affinity and adsorption to the stationary phase determine its rate of movement. This technique can purify materials on both small and large scales for use in experiments. It has applications like isolating active ingredients, separating compound mixtures, drug estimation in formulations, and removing impurities.

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    Column chromatography is a technique used to separate chemical compounds dissolved in a fluid mixture. It works by differential adsorption of compounds to the stationary phase material in the column, causing the compounds to move through the column at different rates and elute separately. Factors like a compound's affinity and adsorption to the stationary phase determine its rate of movement. This technique can purify materials on both small and large scales for use in experiments. It has applications like isolating active ingredients, separating compound mixtures, drug estimation in formulations, and removing impurities.

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
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    139 views11 pages

    Column Chromatography PDF

    Uploaded by

    GLADYS FERNANDES
    Column chromatography is a technique used to separate chemical compounds dissolved in a fluid mixture. It works by differential adsorption of compounds to the stationary phase material in the column, causing the compounds to move through the column at different rates and elute separately. Factors like a compound's affinity and adsorption to the stationary phase determine its rate of movement. This technique can purify materials on both small and large scales for use in experiments. It has applications like isolating active ingredients, separating compound mixtures, drug estimation in formulations, and removing impurities.

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    of 11

    Separation

    Techniques-III
    Column
    Chromatography
    Column Chromatography
    In chemistry, Column chromatography is a technique which is used to separate a
    single chemical compound from a mixture dissolved in a fluid.

    It separates substances based on differential adsorption of compounds to the


    adsorbent as the compounds move through the column at different rates which
    allow them to get separated in fractions.

    This technique can be used on small scale as well as large scale to purify
    materials that can be used in future experiments.

    This method is a type of adsorption chromatography technique.


    Column Chromatography- Principle
    When the mobile phase along with the mixture that needs to be separated is
    introduced from the top of the column, the movement of the individual
    components of the mixture is at different rates. The components with lower
    adsorption and affinity to stationary phase travel faster when compared to the
    greater adsorption and affinity with the stationary phase. The components that
    move fast are removed first whereas the components that move slow are eluted
    out last.

    The adsorption of solute molecules to the column occurs in a reversible manner.


    The rate of the movement of the components is expressed as:

    Rf = the distance travelled by solute/ the distance travelled by solvent

    Rf is the retardation factor.


    Column Chromatography- Diagram
    Column Chromatography- Procedure
    Mobile phase – This phase is made up of solvents and it performs the following
    functions:

    It acts as a solvent – sample mixture can be introduced in the column.

    It acts as a developing agent – helps in the separation of components in the


    sample to form bands.

    It acts as an eluting agent – the components that are separated during the
    experiment are removed from the column

    Some examples of solvents used as mobile phase based on their polarity are –
    ethanol, acetone, water, acetic acid, pyridine, etc.
    Stationary phase – It is a solid material which should have good
    adsorption property and meet the conditions given below:

    Shape and size of particle: Particles should have uniform shape and
    size in the range of 60 – 200μ in diameter.

    Stability and inertness of particles: high mechanical stability and


    chemically inert. Also, no reaction with acids or bases or any other
    solvents used during the experiment.

    It should be colourless, inexpensive and readily available.

    Should allow free flow of mobile phase

    It should be suitable for the separation of mixtures of various


    compounds.
    Column Chromatography- Applications
    Column Chromatography is used to isolate active ingredients.

    It is very helpful in Separating compound mixtures.

    It is used to determine drug estimation from drug formulations

    It is used to remove impurities.

    Used to isolation metabolites from biological fluids.


    Types Of Column Chromatography

    Adsorption column
    chromatography –
    Adsorption
    chromatography is a
    technique of separation, in
    which the components of
    the mixture are adsorbed
    on the surface of the
    adsorbent.
    2.Partition column chromatography –
    The stationary phase, as well as mobile
    phase, are liquid in partition
    chromatography.
    3.Gel column chromatography –
    In this method of chromatography,
    the separation takes place through a
    column packed with gel. The
    stationary phase is a solvent held in
    the gap of a solvent.
    4.Ion exchange column
    chromatography- A chromatography
    technique in which the stationary
    phase is always ion exchange resin.

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