Consider that you wish to separate a mixture of arginine, glutamate and valine by ion exhange chromatography .
Using a cation exchange column at pH 6 , which amino acid would you expect to elute first and which last?
pKa and pI values are as follows:
Arginine: p K 1 = 2.17, p K 2 = 9.04, p K R = 12.48, pI = 10.76
Glutamate: p K 1 = 2.19, p K 2 = 9.67, p K R = 4.25, pI = 3.22
Valine: p K 1 = 2.32, p K 2 = 9.62, pI = 5.97

Question

Consider that you wish to separate a mixture of arginine, glutamate and valine by  ion exhange chromatography . 
 Using a  cation exchange column at pH 6 , which amino acid would you expect to elute first and which last? 
 pKa and pI values are as follows: 
 Arginine:  p K 1  = 2.17, p K 2  = 9.04, p K R  = 12.48, pI = 10.76 
 Glutamate:  p K 1  = 2.19, p K 2  = 9.67, p K R  = 4.25, pI = 3.22 
 Valine:  p K 1  = 2.32, p K 2  = 9.62, pI = 5.97

Answer

Glutamate first. Valine last.

Answer

Arginine first. Valine last.

Answer

Arginine first. Glutamate last.

Answer

Glutamate first. Arginine last.

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