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    (실험4) Chromatography

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    smileban859
    The document describes chromatography techniques used to separate mixtures by polarity. It details the theory, apparatus, procedure, and results of using thin-layer chromatography to separate pigments based on their polarity and calculate Rf values.

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    (실험4) Chromatography

    Uploaded by

    smileban859
    5 views11 pages
    The document describes chromatography techniques used to separate mixtures by polarity. It details the theory, apparatus, procedure, and results of using thin-layer chromatography to separate pigments based on their polarity and calculate Rf values.

    Original Description:

    general chemical experiment [4]

    Original Title

    [실험4] Chromatography

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    The document describes chromatography techniques used to separate mixtures by polarity. It details the theory, apparatus, procedure, and results of using thin-layer chromatography to separate pigments based on their polarity and calculate Rf values.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    5 views11 pages

    (실험4) Chromatography

    Uploaded by

    smileban859
    The document describes chromatography techniques used to separate mixtures by polarity. It details the theory, apparatus, procedure, and results of using thin-layer chromatography to separate pigments based on their polarity and calculate Rf values.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    of 11

    Chromatography

    Content
    • Purpose
    • Theory
    • Apparatus and Reagents
    • Procedure
    • Caution
    • Results
    Purpose
    • Learning the basic principle of chromatography (used to
    determine the makeup of a mixed solution) and the concept of
    polarity through the separation of pigments using normal and
    reversed-phase chromatography.

    • Key Concepts
    1. Chromatography
    -Partition Chromatography
    -Thin-Layer Chromatography
    -Normal-Phase Chromatography
    -Reversed-Phase Chromatography
    2. Polarity
    3. Electronegativity
    Theory
     Stages of Chromatography

    Stationary Phase the substance is fixed in place for the


    chromatography procedure
    Mobile Phase the phase where the substance moves
    in a definite direction

     The Basic Principle of Chromatography


    Chromatography is a physical method of separation in which
    the components to be separated are distributed between two
    phases, one of which is stationary (the stationary phase) while the
    other (the mobile phase) moves in a definite direction.
    Theory
     Techniques

    Chromatographic Physical States of Separation


    Bed Shapes Mobile Phase Mechanisms
    - Column Chromatography - Gas Chromatography - Ion Exchange
    - Liquid Chromatography Chromatography
    - Paper Chromatography - Size Exclusion Chro-
    - Planar Chromatography matography
    - Thin-Layer Chromatog-
    raphy

     Retention Time
    Retention time is the characteristic time it takes for a
    particular analyte to pass through the system under set
    conditions.
    Example of GC
    Theory
     Rf value (rate of flow)
    The retention factor, or Rf, is defined as the distance
    traveled by the compound divided by the distance
    traveled by the solvent.

     An Rf value is characteristic for any given compound (provided that the


    same stationary and mobile phases are used). It can provide
    corroborative evidence as to the identity of a compound.
    Apparatus and Reagents
    • 50 mL-Beaker; watch glass

    • TLC plate (4cm x 5cm of silica gel)

    • Capillary tube

    • Pasteur pipette; tweezers

    • Mobile phase (1-butanol:acetic acid:water = 60:15:25)

    • Sample (mixture of red #40, yellow #5 and blue #1)


    Procedure
    1. A small spot of solution containing the sample is applied to a plate, about 1cm
    from the base and left to dry. (sample: a. red #40
    b. yellow #5 c. blue #1 d. unknown)

    2. The plate is dipped into the solvent (a mixture of 1-butanol, acetic acid, DI water
    in the ratio of 60:15:25), placed in a container and sealed with a watch glass.

    3. Once the solvent has reached the line 1cm from the top of the TLC plate, remove
    the plate from the solution and dry it.

    4. Measure the Rf value of the unknown samples and compare with other pure
    pigments. Using this information you can determine which pigments are
    contained in the unknown sample and whether a reaction (if any) is in process or
    has completed.
    Caution
     Prevent evaporation of the mobile phase solution.

     Keep the 1-butanol away from open flames. It is flammable.

     When applying the sample to the TLC, try to make spot as small as
    possible, and apply it repeatedly if it is not dark enough.

     Do not immerse the spot in the mobile phase.


    Result
    • the Distance of the Mobile Phase Solution : _3.0__cm

    • Calculated Rf Value
    Red Yellow Blue #1
    #40 #5 #1
    Distance 2.2 2.5 1.9 1.5/2.2/2.5
    (cm)
    Rf
    value

    • Characterization of the Unknown


    #1:

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