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From a carboxymethyl-cellulose column at pH 6.0, Arginine, valine and Glutamic acid will elute in the order:
- a)Arginine, valine, Glutamic acid
- b)Arginine, Glutamic acid, valine
- c)Glutamic acid, Arginine, valine
- d)Glutamic acid, valine, Arginine
Correct answer is option 'D'. Can you explain this answer?
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From a carboxymethyl-cellulose column at pH 6.0, Arginine, valine and ...
Elution order of Arginine, Valine, and Glutamic acid from carboxymethyl-cellulose column at pH 6.0
Carboxymethyl-cellulose (CMC) column is a type of ion exchange chromatography in which anion exchange resins interact with positively charged molecules. The elution order of amino acids from CMC column depends on their pKa values and the pH of the buffer used. At pH 6.0, the carboxyl groups on CMC resin are partially ionized, and the amino acids are in their zwitterionic form.
The pKa values of Arginine, Valine, and Glutamic acid are as follows:
- Arginine: pKa1=2.17, pKa2=9.04, pKa3=12.48
- Valine: pKa1=2.32, pKa2=9.62
- Glutamic acid: pKa1=2.19, pKa2=4.25, pKa3=9.67
Based on the pKa values and the pH of the buffer, the following elution order is expected:
- Glutamic acid (pI=3.22) will elute first because it has the lowest pI value among the three amino acids and will be negatively charged in the column.
- Valine (pI=6.01) will elute next because it is neutral at pH 6.0 and does not interact strongly with the CMC resin.
- Arginine (pI=10.76) will elute last because it has the highest pI value and will be positively charged in the column, interacting strongly with the negatively charged CMC resin.
Therefore, the correct elution order of Arginine, Valine, and Glutamic acid from CMC column at pH 6.0 is:
- Glutamic acid, Valine, Arginine
Carboxymethyl-cellulose (CMC) column is a type of ion exchange chromatography in which anion exchange resins interact with positively charged molecules. The elution order of amino acids from CMC column depends on their pKa values and the pH of the buffer used. At pH 6.0, the carboxyl groups on CMC resin are partially ionized, and the amino acids are in their zwitterionic form.
The pKa values of Arginine, Valine, and Glutamic acid are as follows:
- Arginine: pKa1=2.17, pKa2=9.04, pKa3=12.48
- Valine: pKa1=2.32, pKa2=9.62
- Glutamic acid: pKa1=2.19, pKa2=4.25, pKa3=9.67
Based on the pKa values and the pH of the buffer, the following elution order is expected:
- Glutamic acid (pI=3.22) will elute first because it has the lowest pI value among the three amino acids and will be negatively charged in the column.
- Valine (pI=6.01) will elute next because it is neutral at pH 6.0 and does not interact strongly with the CMC resin.
- Arginine (pI=10.76) will elute last because it has the highest pI value and will be positively charged in the column, interacting strongly with the negatively charged CMC resin.
Therefore, the correct elution order of Arginine, Valine, and Glutamic acid from CMC column at pH 6.0 is:
- Glutamic acid, Valine, Arginine
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From a carboxymethyl-cellulose column at pH 6.0, Arginine, valine and Glutamic acid will elute in the order:a)Arginine, valine, Glutamic acidb)Arginine, Glutamic acid, valinec)Glutamic acid, Arginine, valined)Glutamic acid, valine, ArginineCorrect answer is option 'D'. Can you explain this answer?
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