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    Name-Avneet Kaur Student No. - 301095622 Post-Lab Questions: Ques 1 - Explain Basic Principle of Paper Chromatography?

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    Adv Sandeep Singh
    This document contains the results of a student's post-lab questions for an experiment on paper chromatography. The student explains that paper chromatography separates chemical substances based on their differing migration speeds on chromatographic paper. Aspartic acid moves slower than leucine or phenylalanine because the nonpolar solvent attracts the nonpolar leucine more than the polar aspartic acid. The data collected showed that leucine had the highest Rf value, indicating it traveled farther on the paper. Ninhydrin and other tests were used to identify the amino acids.

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    Name-Avneet Kaur Student No. - 301095622 Post-Lab Questions: Ques 1 - Explain Basic Principle of Paper Chromatography?

    Uploaded by

    Adv Sandeep Singh
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    This document contains the results of a student's post-lab questions for an experiment on paper chromatography. The student explains that paper chromatography separates chemical substances based on their differing migration speeds on chromatographic paper. Aspartic acid moves slower than leucine or phenylalanine because the nonpolar solvent attracts the nonpolar leucine more than the polar aspartic acid. The data collected showed that leucine had the highest Rf value, indicating it traveled farther on the paper. Ninhydrin and other tests were used to identify the amino acids.

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    This document contains the results of a student's post-lab questions for an experiment on paper chromatography. The student explains that paper chromatography separates chemical substances based on their differing migration speeds on chromatographic paper. Aspartic acid moves slower than leucine or phenylalanine because the nonpolar solvent attracts the nonpolar leucine more than the polar aspartic acid. The data collected showed that leucine had the highest Rf value, indicating it traveled farther on the paper. Ninhydrin and other tests were used to identify the amino acids.

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    © All Rights Reserved
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    Download as docx, pdf, or txt
    70 views2 pages

    Name-Avneet Kaur Student No. - 301095622 Post-Lab Questions: Ques 1 - Explain Basic Principle of Paper Chromatography?

    Uploaded by

    Adv Sandeep Singh
    This document contains the results of a student's post-lab questions for an experiment on paper chromatography. The student explains that paper chromatography separates chemical substances based on their differing migration speeds on chromatographic paper. Aspartic acid moves slower than leucine or phenylalanine because the nonpolar solvent attracts the nonpolar leucine more than the polar aspartic acid. The data collected showed that leucine had the highest Rf value, indicating it traveled farther on the paper. Ninhydrin and other tests were used to identify the amino acids.

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    Name- Avneet Kaur

    Student no.- 301095622

    Post-lab questions:

    Ques 1- Explain basic principle of paper chromatography?

    Ans- Chromatography system that utilizations paper sheets or strips as the adsorbent being the
    stationary stage through which a solution is made to pass is called paper chromatography. It is a
    modest technique for isolating disintegrated chemical substances by their distinctive movement
    rates over the pieces of paper.

    Ques 2- Aspartic acid moves slower on the chromatographic paper than either leucine or
    phenylalanine. How do you explain the difference?

    Ans – the mix solvent used was primarily nonpolar, as a result, nonpolar amino acids like leucine
    are eluting faster than polar amino acid like aspartic acid.

    Discussion

    Chromatography is a technique for separating isolating chemical substances by their different


    migration speeds on the paper causing them to separate. In paper chromatography, the stationary
    phase is a very uniform absorbent paper. The mobile phase is a appropriate liquid solvent or
    mixture of solvents. The separation of chemical substances is based on differential partitioning
    between the mobile and stationary phase. Slight differences in a compound’s separation
    coefficient result in different retention on the stationary phase and thus affect the separation.

    The collected data contains the solvent distance and spot distance. In this experiment, four amino
    acids alanine, aspartic acid, leucine and lysine was used. In the known component’s leucine is
    having the highest Rf value, which is 0.61, Whereas Lysine has the lowest Rf value which is 0.17.
    Ninhydrin was used for detecting the amino acids, since amino acids are colourless compounds.
    Ninhydrin act as a powerful oxidizing agent which reacts with α- amino acids that results in
    purple coloured spots which is termed as Rheuman's purple.

    Rf = distance of component travelled / distance of solvent


    The aspartic acid moves slower than the leucine as it is attracted more to paper than hydrophobic
    solvent, thus has low Rf. In this case the most similar Rf value was taken to be the unknown.
    From the results obtained one can conclude that unknown was most probably [CITATION UKE20 \l
    4105 ].

    Xanthoproteic test was performed which shows presence or absence of phenyl group. It detected
    the protein and undergo nitration reaction with aromatic compound to give yellow color.
    Albumin, casein, tryptophan and tyrosine were positive, and gelatin gave negative reaction for
    xanthoproteic test[ CITATION Cernd \l 4105 ].

    Hopkins-Cole test was performed which shows presence of indole. Indole reacts with glyoxylic
    acid and form purple ring in solution. Albumin, Casein, and tryptophan were positive for
    Hopkins-Cole test. A purple ring appears between the two layers if the test is positive [ CITATION
    Athnd \l 4105 ]

    References

    Athirah, N. (n.d.). LAB BCM AMINO ACID. Retrieved january 29, 2020, from
    https://www.academia.edu/38927279/LAB_BCM_AMINO_ACID

    Cerna, E. (n.d.). Experiment 6. Retrieved january 29, 2020, from


    https://www.academia.edu/13753057/Experiment_6

    CH222 Biochemistry 1 lab manual (2020) ecentennial. Separation and identification of amino
    acids, centennial college, Scarborough, Ontario. Retrieved January 29, 2020, from

    https://e.centennialcollege.ca/d2l/le/content/504972/viewContent/5316131/View
    UKEssays. (n.d.). Separation of Amino Acids by Paper Chromatography. Retrieved january 29,
    2020, from https://www.ukessays.com/essays/chemistry/separation-amino-acids-paper-
    7414.php

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