Paper chromatography | Principle | Procedure | Development techniques | Applications

Bio science
11 Nov 201915:12
EducationalLearning
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TLDRThis educational video offers a comprehensive guide to paper chromatography, a technique for separating molecules using paper as the stationary phase and a solvent as the mobile phase. It explains the process, including the preparation of materials, application of the sample, and observation of substance migration. The video also covers the calculation of the RF value, different development techniques, and the applications of paper chromatography in separating and analyzing organic and inorganic compounds, amino acids, and halide ions.

Takeaways
  • πŸ” Paper chromatography is a technique used for the separation of molecules from a sample using paper as the stationary phase.
  • πŸ“ Chromatography involves both a stationary and a mobile phase, with the type of stationary phase depending on the technique used.
  • πŸ“‘ The materials required for paper chromatography include a stationary phase (filter paper), a mobile phase (solvent), and a sample.
  • πŸ“ The paper is marked with a start point and an endpoint, and the sample is placed at the start point for the chromatography process.
  • 🌊 The paper is then dipped into the mobile phase, ensuring only the sample area is submerged in the solvent.
  • πŸ”¬ The migration of substances is observed, which depends on the size, nature of the sample, and the KD value.
  • πŸ“ˆ The RF value, or Retention Factor, is calculated using the distance traveled by the solute from the origin divided by the distance traveled by the solvent from the origin.
  • πŸ”„ One-dimensional and two-dimensional techniques are mentioned, with the two-dimensional technique being more effective for proper separation and less diffusion.
  • πŸ“ˆ The type of paper used is Whatman number one filter paper, which is preferred for its purity and composition.
  • πŸ§ͺ The preparation of the paper involves cutting, washing, and drying to remove impurities.
  • 🌑 After the chromatography process, the paper is dried and then exposed to UV light or potassium permanganate solution to visualize the separated particles.
Q & A

  • What is the main purpose of chromatography?

    -The main purpose of chromatography is to separate molecules from a sample based on their properties.

  • What is paper chromatography?

    -Paper chromatography is a type of chromatography that uses paper as the stationary phase to separate the components of a mixture.

  • What are the two main phases involved in chromatography?

    -The two main phases involved in chromatography are the stationary phase and the mobile phase.

  • What type of paper is recommended for paper chromatography?

    -Whatman number one filter paper is recommended for paper chromatography due to its composition and suitability for separation.

  • What should be done to prepare the paper for chromatography?

    -The paper should be cut to an appropriate size and shape, washed to remove impurities, and then dried before use in the experiment.

  • How should the sample be applied on the paper in paper chromatography?

    -The sample should be applied at the start point on the paper using a capillary tube, ensuring it is in microgram quantity and not in excess.

  • What is the role of the solvent in the mobile phase?

    -The solvent in the mobile phase helps in the migration of the sample particles and facilitates the separation process in chromatography.

  • What is the significance of the RF value in paper chromatography?

    -The RF value, or the Retention Factor, is used to quantify the distance a solute travels relative to the solvent front, which helps in identifying and comparing different solutes.

  • What are the two main types of techniques used in paper chromatography?

    -The two main types of techniques used in paper chromatography are the one-dimensional technique and the two-dimensional technique.

  • Why is it preferable to use the two-dimensional technique over the one-dimensional technique in paper chromatography?

    -The two-dimensional technique is preferable because it provides better separation of molecules with less diffusion, leading to more accurate results compared to the one-dimensional technique.

  • What are some applications of paper chromatography?

    -Applications of paper chromatography include the separation and purification of organic and inorganic compounds, analysis of amino acids, identification of halide ions, and separation of carbohydrates and pigments.

Outlines
00:00
πŸ” Introduction to Paper Chromatography

This paragraph introduces the topic of paper chromatography, a technique used for the separation of molecules within a sample. It emphasizes the importance of understanding the principle behind chromatography and the different types available, with a focus on paper chromatography. The stationary phase in this method is paper, while the mobile phase is a solvent. The paragraph outlines the materials needed, such as the specific type of filter paper and the sample, and describes the initial steps of the technique, including marking the paper with start and endpoints and applying the sample. It also touches on the importance of the KD value and the RF value in observing the migration of substances during the process.

05:01
πŸ“š Techniques and Materials in Paper Chromatography

The second paragraph delves deeper into the technical aspects of paper chromatography, discussing the types of techniques used, including one-dimensional and two-dimensional methods. It highlights the superiority of the two-dimensional technique for achieving better separation and less diffusion of molecules. The paragraph also covers the selection of materials, emphasizing the use of Whatman number one filter paper due to its purity and suitability for chromatography. It explains the preparation of the paper, solute mixture, and the spotting process. Additionally, it discusses the choice of solvent, which can be polar, nonpolar, or moderately polar, depending on the sample. The development techniques, such as ascending, descending, and radial methods, are also briefly introduced.

10:02
🌐 Advanced Chromatography Techniques and Applications

The final paragraph discusses the formation and use of different development methods in chromatography, including the ascending, descending, and radial techniques. It explains how the direction of particle migration is influenced by the position of the mobile phase. The paragraph also addresses the post-chromatography steps, such as removing and drying the paper, and using potassium permanganate solution or UV light for better observation of the separated molecules. It concludes with the applications of paper chromatography, including the separation and identification of organic and inorganic compounds, amino acids, halide ions, and carbohydrates. The video script ends with an invitation for viewers to join a WhatsApp group for further notes and discussions on paper chromatography.

Mindmap
Keywords
πŸ’‘Paper Chromatography
Paper chromatography is a method within the field of chromatography used for separating the components of a mixture based on their affinity to the paper and the solvent. It is a fundamental technique in analytical chemistry and is central to the video's theme. In the script, it is described as a process where the separation of mixture components occurs using paper as a stationary phase.
πŸ’‘Chromatography
Chromatography is a collective term for a family of methods used to separate mixtures into their individual components. It is the overarching concept in the video, with paper chromatography being one of its types. The script explains that chromatography techniques are used for the separation of molecules from a sample.
πŸ’‘Stationary Phase
In the context of chromatography, the stationary phase is the medium on which the sample is applied and through which the mobile phase travels. In the video script, the stationary phase is specifically the paper used in paper chromatography, which is marked with start and end points for the application of the sample and observation of the separation process.
πŸ’‘Mobile Phase
The mobile phase in chromatography is the substance that moves through the stationary phase, carrying the sample components with it. The script mentions that the mobile phase is a solvent, which is common across all types of chromatography techniques, and it is responsible for the migration of substances in paper chromatography.
πŸ’‘Sample
The sample in chromatography is the mixture that is to be separated into its individual components. The script specifies that the sample should be applied to the start point on the paper in paper chromatography and that its migration depends on its size, nature, and the KD value.
πŸ’‘RF Value
The RF value, or the Retention Factor, is a measure used in chromatography to quantify the interaction between the solute and the stationary phase. It is calculated as the distance traveled by the solute divided by the distance traveled by the solvent from the origin. The script explains that the RF value is crucial for understanding the separation efficiency in paper chromatography.
πŸ’‘One-Dimensional Technique
A one-dimensional technique in paper chromatography refers to the process where the stationary phase is presented in a vertical position and the separation occurs in one direction. The script contrasts this with the two-dimensional technique, noting that one-dimensional is less commonly used due to potential issues with diffusion and incomplete separation.
πŸ’‘Two-Dimensional Technique
The two-dimensional technique in paper chromatography involves the stationary phase being tilted or placed in a different orientation, allowing for a second dimension of separation. The script highlights that this technique is more effective, resulting in less diffusion and better separation of molecules.
πŸ’‘Ascending Technique
The ascending technique in chromatography is a development method where the mobile phase moves against the gravitational force, typically moving upward. The script describes this as a method where the particles' migration moves against the gravitational force, which is an important aspect of the development process in paper chromatography.
πŸ’‘Descending Technique
The descending technique is another development method in chromatography, where the mobile phase moves in the direction of the gravitational force, causing the sample to move downward. The script mentions this technique as one that is less commonly used in paper chromatography due to its limitations.
πŸ’‘Radial Development Method
The radial development method is a specific technique in chromatography where the sample is placed in the center of a circular stationary phase, and the mobile phase moves radially outward. The script describes this method as being used in rare cases and results in a circular pattern of separated molecules.
πŸ’‘Whatman Number One Filter Paper
Whatman Number One filter paper is a specific type of paper used in paper chromatography due to its high purity and uniformity. The script emphasizes its use because it contains alpha cellulose, beta cellulose, and other components that facilitate better separation of compounds.
πŸ’‘Visualizing Agent
A visualizing agent is a substance used in chromatography to make the separated components visible after the development process. The script mentions potassium permanganate and silver nitrate as examples of visualizing agents used for observing the separated particles and identifying halide ions, respectively.
Highlights

Introduction to paper chromatography, including a complete detailed explanation and its applications.

Explanation of the principle of chromatography and its use for separating molecules from a sample.

Description of paper chromatography, where paper is used as the stationary phase.

Materials required for paper chromatography: stationary phase (paper), mobile phase (solvent), and sample.

Step-by-step process: Marking the start and end points on the paper.

Placing the sample on the start point of the paper.

Dipping the stationary phase (paper) into the mobile phase (solvent).

Observation of the migration of substances, dependent on sample size, nature, and KD value.

Calculation of the RF value: Distance traveled by the solute divided by distance traveled by the solvent.

Comparison of one-dimensional and two-dimensional paper chromatography techniques.

Advantages of two-dimensional technique: Better separation of molecules and reduced diffusion.

Details on the preparation of the paper and the importance of using Whatman number one filter paper.

Explanation of different development techniques: ascending, descending, and radial.

Steps involved in drying and observing the paper after chromatography, using UV light or potassium permanganate solution.

Applications of paper chromatography: Separation of organic and inorganic compounds, analysis of amino acids, identification of halide ions, and separation of carbohydrates and pigments.

Transcripts

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