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    Thin Layer Chromatography

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    GLADYS FERNANDES
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    Thin Layer Chromatography

    Uploaded by

    GLADYS FERNANDES
    91 views12 pages
    A power point presentation on Thin Layer Chromatography

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    © © All Rights Reserved

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    PDF, TXT or read online from Scribd

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    A power point presentation on Thin Layer Chromatography

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    91 views12 pages

    Thin Layer Chromatography

    Uploaded by

    GLADYS FERNANDES
    A power point presentation on Thin Layer Chromatography

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 12

    Separation Techniques-II

    Thin layer chromatography


    Thin layer chromatography
    Thin Layer Chromatography is a technique used to isolate non-volatile mixtures.

    The experiment is conducted on a sheet of aluminium foil, plastic, or glass which


    is coated with a thin layer of adsorbent material.

    The material usually used is aluminium oxide, cellulose, or silica gel.

    On completion of the separation, each component appears as spots separated


    vertically.Each spot has a retention factor (Rf) expressed as:

    Rf = dist. travelled by sample / dist. travelled by solvent


    The factors affecting retardation factor are the solvent system, amount of material
    spotted, absorbent and temperature.

    TLC is one of the fastest, least expensive, simplest and easiest chromatography
    technique.
    Thin Layer Chromatography -Principle
    The compounds in the mobile phase move over the surface of the stationary
    phase. The movement occurs in such a way that the compounds which have a
    higher affinity to the stationary phase move slowly while the other compounds
    travel fast. Therefore, the separation of the mixture is attained.
    Diagram of thin layer chromatography
    Thin layer chromatography-Procedure
    1.Thin Layer Chromatography Plates –ready-made plates are used which are
    chemically inert and stable. The stationary phase is applied on its surface in the
    form of a thin layer. The stationary phase on the plate has a fine particle size and
    also has a uniform thickness.

    2.Thin Layer Chromatography Chamber – Chamber is used to develop plates. It


    is responsible to keep a steady environment inside which will help in developing
    spots. Also, it prevents the solvent evaporation and keeps the entire process
    dust-free.
    3.Thin Layer Chromatography Mobile phase – Mobile phase is the one that
    moves and consists of a solvent mixture or a solvent. This phase should be
    particulate-free. The higher the quality of purity the development of spots is better.

    4 .Thin Layer Chromatography Filter Paper – It has to be placed inside the


    chamber. It is moistened in the mobile phase.
    Thin layer chromatography- Application
    The qualitative testing of Various
    medicines such as sedatives, local
    anaesthetics, anticonvulsant
    tranquilisers, analgesics,
    antihistamines, steroids, hypnotics
    is done by TLC.
    TLC is extremely useful in Biochemical analysis
    such as separation or isolation of biochemical
    metabolites from its blood plasma, urine, body
    fluids, serum, etc.
    Thin layer chromatography can be used to
    identify natural products like essential oils or
    volatile oil, fixed oil, glycosides, waxes,
    alkaloids, etc
    It is used in the food industry, to separate and
    identify colours, sweetening agent, and
    preservatives

    It is used in the cosmetic industry.

    It is used to study if a reaction is complete.


    Disadvantage Of Thin Layer Chromatography
    Thin Layer Chromatography plates do not have longer stationary phase.

    When compared to other chromatographic techniques the length of separation is


    limited.

    The results generated from TLC are difficult to reproduce.

    Since TLC operates as an open system, some factors such as humidity and
    temperature can be consequences to the final outcome of the chromatogram.

    The detection limit is high and therefore if you want a lower detection limit, you
    cannot use TLC.

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