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    Plant Pigment Chromatography

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    jwalantkbhatt
    This document describes using paper chromatography to separate plant pigments from spinach leaves. Pigments like chlorophyll, beta carotene, and xanthophyll can be separated based on how far they travel up chromatography paper via capillary action using a solvent. Beta carotene travels furthest as it is highly soluble and does not form hydrogen bonds with the paper. Xanthophyll travels less far as it forms hydrogen bonds. Chlorophyll travels the shortest distance as it is most tightly bound to the paper. The procedure involves extracting pigments from spinach leaves onto paper and developing the bands using a solvent to calculate Rf values to identify the pigments.

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    Plant Pigment Chromatography

    Uploaded by

    jwalantkbhatt
    56 views2 pages
    This document describes using paper chromatography to separate plant pigments from spinach leaves. Pigments like chlorophyll, beta carotene, and xanthophyll can be separated based on how far they travel up chromatography paper via capillary action using a solvent. Beta carotene travels furthest as it is highly soluble and does not form hydrogen bonds with the paper. Xanthophyll travels less far as it forms hydrogen bonds. Chlorophyll travels the shortest distance as it is most tightly bound to the paper. The procedure involves extracting pigments from spinach leaves onto paper and developing the bands using a solvent to calculate Rf values to identify the pigments.

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    This document describes using paper chromatography to separate plant pigments from spinach leaves. Pigments like chlorophyll, beta carotene, and xanthophyll can be separated based on how far they travel up chromatography paper via capillary action using a solvent. Beta carotene travels furthest as it is highly soluble and does not form hydrogen bonds with the paper. Xanthophyll travels less far as it forms hydrogen bonds. Chlorophyll travels the shortest distance as it is most tightly bound to the paper. The procedure involves extracting pigments from spinach leaves onto paper and developing the bands using a solvent to calculate Rf values to identify the pigments.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    56 views2 pages

    Plant Pigment Chromatography

    Uploaded by

    jwalantkbhatt
    This document describes using paper chromatography to separate plant pigments from spinach leaves. Pigments like chlorophyll, beta carotene, and xanthophyll can be separated based on how far they travel up chromatography paper via capillary action using a solvent. Beta carotene travels furthest as it is highly soluble and does not form hydrogen bonds with the paper. Xanthophyll travels less far as it forms hydrogen bonds. Chlorophyll travels the shortest distance as it is most tightly bound to the paper. The procedure involves extracting pigments from spinach leaves onto paper and developing the bands using a solvent to calculate Rf values to identify the pigments.

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    Plant Pigment Chromatography

    Introduction: Paper chromatography is a technique used to separate substances in


    a mixture based on the movement of the different substances up a piece of paper
    by capillary action. Pigments extracted from plant cells contain a variety of
    molecules, such as chlorophylls, beta carotene, and xanthophyll, that can be
    separated using paper chromatography. A small sample of plant pigment placed on
    chromatography paper travels up the paper due to capillary action. Beta carotene is
    carried the furthest because it is highly soluble in the solvent and because tit forms
    no hydrogen bonds with the chromatography paper fibers. Xanthophyll contains
    oxygen and does not travel quite as far with the solvent because it is less soluble the
    beta carotene and forms hydrogen bonds with the paper. Chlorophylls are bound more
    tightly to the paper than the other two, so they travel the shortest distance.

    Objectives:
    • Separate plant pigments in Spinach leaves
    • Calculate the Rf values for the pigments

    Materials
    100 mL graduated cylinder Aluminum foil
    Chromatography paper Pencil Spinach leaves
    Scissors Coin Chromatography Solvent
    Ruler

    Procedure
    The solvent in this experiment is flammable and poisonous. Be sure there are no open flames in the lab during
    the experiment. Avoid inhaling the fumes. Close the solvent bottle immediately after you are finished pouring the
    solvent. Notify your teacher is any accidents occur.

    1. Pour 10 mL of chromatography solvent into a 100 mL graduated cylinder. Cover the cylinder with aluminum foil.

    2. Cut the end of the chromatography paper into a point.

    3. Draw a pencil line 2.0 cm above the pointed end of the paper.

    4. Use the coin to extract pigments from the spinach leaf. Place a small amount of
    the leaf on top of the pencil line. Use the edge of the coin to push the plant cells
    into the chromatography paper. Repeat the procedure 10 times making sure to
    use a different part of the leaf each time.

    5. Place the chromatography paper in the cylinder so that the pointed end just
    touches the solvent. Make are the pigment is not in the solvent.

    6. Fold over the paper at the top of the cylinder and recover with aluminum foil.
    Wait until the solvent is approximately 2 cm from the top of the paper. Remove the paper and mark the solvent front before
    it evaporates.

    7. Return the solvent to the stock bottles.


    8. Allow the paper to dry. Mark the bottom of each pigment band. Measure the distance each pigment moved from the
    starting line to the bottom of the pigment band. Record the distance of each pigment and the distance the solvent moved, in
    millimeters.

    9. Identify each of the bands and label them on the chromatography paper.
    • Beta carotene: yellow to yellow orange
    • Xanthophyll: yellow
    • Chlorophyll a: bright green to blue green
    • Chlorophyll b: yellow green to olive green

    10. Cut the chromatography paper in half length wise and tape into your
    lab notebook.

    DATA: Sample Data Tables


    Table 1
    Band Number Distance Traveled (mm) Band Color Identity
    1
    2
    3
    4
    5*
    Distance solvent traveled (mm)
    * The fifth band may not appear

    Table 2
    Molecule Rf
    Beta carotene
    Xanthophyll
    Chlorophyll a
    Chlorophyll b

    The retention factor (Rf) may be defined as the ratio of the distance traveled by the solvent to the distance traveled by the
    solvent. Rf values are usually expressed as a fraction of 2 decimal places. If the Rf value is zero, the solute is immobile and not
    soluble in the solvent. A known Rf value in a given solvent can be used to identify a molecule.

    𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑 𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡 𝑏𝑏 𝑝𝑝𝑝𝑝𝑝𝑝𝑝


    𝑅𝑓 =
    𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑 𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡 𝑏𝑏 𝑠𝑠𝑠𝑠𝑠𝑠𝑠

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