Techniques in Biotechnology Chapter 2: Thin Layer Chromatography (TLC)
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Questions and Answers

What is the principle of TLC?

  • The distribution of a compound between a solid phase and a liquid mobile phase (correct)
  • The reaction of a compound with the reagents used for visualization
  • The separation of two compounds based on their boiling points
  • The interaction between a compound and the TLC plate material

    • What happens to a less polar compound in TLC?

  • It has a weaker interaction with the silica
  • It moves higher up the plate (correct)
  • It stays with the mobile phase
  • It has a lower Rf value

    • How are individual separated compounds visualized in TLC?

  • By using reagents like Iodine, Sulfuric acid, Ninhydrin, Potassium Permagnet, Ferric chloride, UV light, etc. (correct)
  • By reacting with the solvent used in TLC
  • By heating the TLC plate in an oven
  • Using UV light only

    • What is used as the solid phase in TLC?

    <p>Silica gel</p> Signup and view all the answers

    How is Rƒ (Retardation Factor) value calculated in TLC?

    <p>By dividing the distance the compound traveled by the distance the solvent traveled</p> Signup and view all the answers

    What is the main purpose of using Thin Layer Chromatography (TLC)?

    <p>To identify compounds present in a given mixture/substance</p> Signup and view all the answers

    What is the role of the stationary phase in Thin Layer Chromatography (TLC)?

    <p>To compete with the mobile phase for binding to the solute</p> Signup and view all the answers

    Which material can be used as the stationary phase in Thin Layer Chromatography (TLC)?

    <p>Cellulose</p> Signup and view all the answers

    What is the mobile phase in Thin Layer Chromatography (TLC)?

    <p>The solvent or solvent mixture drawn up the plate via capillary action</p> Signup and view all the answers

    What can Thin Layer Chromatography (TLC) be used to determine?

    <p>The purity of a substance</p> Signup and view all the answers

    Study Notes

    Principle of TLC

    • The principle of TLC is based on the difference in the partitioning behavior of the components of a mixture between the stationary phase and the mobile phase.

    Separation and Visualization

    • A less polar compound moves faster and farther up the TLC plate, while a more polar compound moves slower and stays closer to the starting point.
    • Individual separated compounds are visualized in TLC by using a chemical reagent or by exposing the plate to iodine vapor or ultraviolet light.

    Stationary Phase

    • The solid phase used in TLC is usually a thin layer of silica gel or alumina on a glass plate or plastic sheet.
    • The stationary phase is responsible for separating the components of a mixture based on their polarity.

    Rƒ (Retardation Factor) Value

    • The Rƒ value is calculated by dividing the distance traveled by the compound by the distance traveled by the solvent front.

    Main Purpose of TLC

    • The main purpose of using TLC is to identify, separate, and quantify the components of a mixture.

    Role of Stationary Phase

    • The role of the stationary phase is to separate the components of a mixture based on their polarity, allowing them to be visualized and characterized.

    Stationary Phase Materials

    • Silica gel, alumina, and cellulose can be used as the stationary phase in TLC.

    Mobile Phase

    • The mobile phase in TLC is a solvent that moves up the plate, carrying the components of the mixture with it.

    Applications of TLC

    • TLC can be used to determine the purity of a substance, identify the components of a mixture, and monitor the progress of a reaction.

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    Description

    Explore the fundamentals of Thin Layer Chromatography (TLC) in the context of Biotechnology Chapter 2. Learn about the principles and applications of TLC and its significance in biotechnological processes.

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