Paper Chromatography - MeitY OLabs

00:05:38
https://www.youtube.com/watch?v=7q5HDMXSdtU

Summary

TLDRThe video explains the procedure of paper chromatography to separate plant pigments using spinach leaves. It describes materials such as fresh spinach leaves, filter paper, chromatography chamber, ether acetone solvent, and more. The method involves cutting the leaves, grinding them with acetone to extract pigments, and applying this on a notched filter paper strip. The strip is then placed in a chromatography chamber with solvent, ensuring the pigment spot stays above the solvent level. As the solvent rises, it separates the pigments into colored bands. Observations include four distinct pigment bands representing different pigments: carotene, xanthophylls, chlorophyll a, and chlorophyll b. The RF value for each pigment can be calculated, ensuring certain precautions like keeping the filter paper's loading spot properly positioned above the solvent. The process highlights separating compounds based on differences in their physical and chemical properties.

Takeaways

  • 🧪 Paper chromatography is used to separate plant pigments.
  • 🌿 Fresh spinach leaves are used for extracting pigments.
  • 📝 Filter paper preparation includes a notched strip and marking with a pencil.
  • 💧 Solvent travels up the paper, separating compounds.
  • 🌈 Pigment bands appear in distinct colors on the paper.
  • 📏 RF value is calculated by comparing distances traveled.
  • 🔍 Carotene is the topmost orange-yellow band.
  • 🟢 Xanthophylls and chlorophylls appear in greenish bands.
  • ⚠️ Proper placement of filter paper in the chamber is crucial.
  • 🍃 Freshness of spinach leaves affects the results.

Timeline

  • 00:00:00 - 00:05:38

    The tutorial explains the concept of paper chromatography, which is used to separate molecules based on differences in their properties. Specifically, this method uses the capillary action to move a solvent through a paper to separate compounds into colored spots. The materials required for this procedure include spinach leaves and various lab equipment. The steps involve preparing an extract from spinach leaves, applying it to filter paper, and allowing a solvent to separate the pigments. Observations and calculations follow, such as measuring the distance of pigment bands and calculating RF values, leading to the identification of pigments like carene and chlorophyll. Precautions emphasize maintaining specified distances to ensure accuracy.

Mind Map

Video Q&A

  • What is paper chromatography?

    It is a technique used to separate molecules on the basis of size, shape, mass, charge, solubility, and absorption properties.

  • What materials are needed for paper chromatography as shown in the video?

    Fresh spinach leaves, filter paper strips, a chromatography chamber, mortar and pestle, ether acetone solvent, measuring cylinder, capillary tube, thread, stapler, scissors, spatula, watch glass, scale, and a pencil.

  • How is the chromatography paper prepared?

    A strip of filter paper is notched, a line is drawn 2-3 cm from the notch, and the pigment extract is applied at the center of the line.

  • What is the role of the solvent in paper chromatography?

    The solvent helps separate the mixture's compounds into colored spots as it flows through the paper by capillary attraction.

  • What observations are made from the chromatography experiment?

    The paper strip shows four distinct pigment bands, each representing different pigments identified by their colors.

  • How do you calculate the RF value in chromatography?

    RF is the distance traveled by the compound divided by the distance traveled by the solvent.

  • What does each pigment band represent?

    The orange-yellow band is carotene, the yellowish band is xanthophylls, the dark green band is chlorophyll a, and the light green band is chlorophyll b.

  • What are the precautions to take during the experiment?

    Ensure spinach leaves are fresh and green. Hang the filter paper strip with the loading spot 1 cm above the solvent level.

  • How do you prepare the plant extract?

    Grind fresh spinach leaves with acetone in a mortar and pestle, then collect the extract using a spatula into a watch glass.

  • What is the function of the capillary tube in this experiment?

    It is used to apply the plant extract onto the filter paper.

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  • 00:00:00
    [Music]
  • 00:00:11
    paper
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    [Music]
  • 00:00:16
    chromatography chomatography is a
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    technique used to separate molecules on
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    the basis of differences in size shape
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    Mass charge solubility and absorption
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    properties
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    in paper chromatography the mixture is
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    spotted on the paper dried and the
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    solvent is allowed to flow along the
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    sheet by capillary
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    attraction as the solvent slowly moves
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    through the paper the different
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    compounds of the mixture separate into
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    different colored spots the paper is
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    dried and the position of the different
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    compounds is
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    visualized different plant segments can
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    be separated
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    by using the technique of paper
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    [Music]
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    chromatography materials
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    required fresh spinach leaves filter
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    paper strips chromatography
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    chamber MTAR and
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    pesel ether acetone solvent measuring
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    cylinder containing 5 ml acetone
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    capillary tube
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    thread
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    stapler
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    scissors
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    spatula watch glass scale and a
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    [Music]
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    pencil
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    [Music]
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    procedure take a few freshly plucked
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    green spinach leaves using scissors cut
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    the spinach leaves into small pieces and
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    let them fall into to the
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    mortar take a measuring cylinder that
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    contains 5 ml of acetone and pour it
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    into the
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    mortar grind the spinach leaves using
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    the mortar and
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    pesel place the extract into a watch
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    glass using a
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    spatula take a strip of filter paper
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    having a narrow Notch at one end of the
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    strip take a pencil and a scale and drw
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    a horizontal line with the pencil about
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    2 to 3 cm away from the tip of the
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    notch put a drop of the pigment extract
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    in the middle of the line with the help
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    of a capillary tube allow the drop to
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    dry and repeat till four or five drops
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    are placed on the
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    paper take the chromatographic chamber
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    and pour ether acetone solvent into
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    it fold one end of the filter paper
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    strip and stap
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    it using a thread hang the filter paper
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    strip into the chromatographic
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    chamber the loading spot should remain
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    about 1 cm above the solvent
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    level leave the chromatographic chamber
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    undisturbed for some time we can observe
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    as the solvent moves through the paper
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    it spreads the different pigments of the
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    mixture in various
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    distances when the solvent rises above
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    34 of the way up the strip remove the
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    strip carefully and let it
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    dry
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    observations the dried chromatographic
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    paper strip shows four distinct pigment
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    bands different pigments can be
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    identified by their
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    colors
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    calculations RF value of each pigment
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    spot can be calculated by the equation
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    RF is equal to distance traveled by the
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    compound divided by distance traveled by
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    the
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    solvent measure the distance of each
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    pigment band from the loading spot and
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    also the distance traveled by the
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    solvent calculate the RF value using the
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    equation and record the values into a
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    table
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    [Music]
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    result the topmost orange yellow band of
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    pigments in the separation corresponds
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    to
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    kerene the yellowish band appearing
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    below it indicates the zantho
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    fils the third from the above dark green
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    band represents chlorophyll
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    a the lower most yellowish green band is
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    that of Chlorophyll
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    [Music]
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    B
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    precautions spinach leaves should be
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    fresh and
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    green the loading spot should be 2 to 3
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    cm away from the tip of the
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    knot while hanging the strips in the
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    chromatography chamber the loading spot
  • 00:05:25
    should remain about 1 cm above the
  • 00:05:28
    solvent level
  • 00:05:31
    [Music]
Tags
  • Chromatography
  • Spinach
  • Pigments
  • Separation
  • Solvent
  • Filter Paper
  • Carotene
  • Chlorophyll
  • Xanthophylls
  • Laboratory Technique