α-AMINO ACID

 The actual structure of amino acid is ionic and depends on the pH. The carboxyl group loses a
proton, giving a carboxylate ion, and the amino group is protonated to an ammonium ion. This
structure is called a dipolar ion or a zwitterion.

 The dipolar nature of amino acids gives them some unusual properties:
1. Amino acids have high melting points, generally over 200 °C.
2. Amino acids are more soluble in water than they are in ether, dichloromethane, and other
common organic solvents.
3. Amino acids have much larger dipole moments than simple amines or simple acids.
4. Amino acids are less acidic than most carboxylic acids and less basic than most amines.
 ZWITTERION
• In an acidic medium, the –COO- group is protonated to a free –COOH group, and the molecule
has an overall positive charge. As the pH is raised, the –COOH loses its proton at about pH 2. This
point is called pKa1, the first acid-dissociation constant.

• In alkaline medium, as the pH is raised further, the –NH3+ group loses its proton at about pH 9 or
10. This point is called pKa2, the second acid-dissociation constant. Above this pH, the molecule
has an overall negative charge.
ZWITTERION
 ISOELECTRONIC POINT (PI)
 The Isoelectronic point also known as iso ionic point is the pH at which the amino acid does not
migrate in an electric field, i.e. it is the pH at which the amino acid is in neutral form (have equal
positive and negative charges), i.e. the zwitterion form is dominant.

 Calculating PI values

For an amino acid with only one amine and one carboxyl group, the pI can be calculated from
the mean of the pKas of the molecule.
 ELECTROPHORESIS
• Electrophoresis is the technique to separate the mixtures of charged amino acids when placed in
an electric field. A streak of the amino acid mixture is placed in the center of a layer of acrylamide
gel or a piece of filter paper wet with a buffer solution. Two electrodes are placed in contact with
the edges of the gel or paper, and a potential of several thousand volts is applied across the
electrodes. Positively charged (cationic) amino acids are attracted to the negative electrode (the
cathode), and negatively charged (anionic) amino acids are attracted to the positive electrode
(the anode). An amino acid at its isoelectric point has no net charge, so it does not move.
For example, consider a mixture of alanine, lysine, and aspartic acid in a
buffer solution at pH 6. Alanine is at its isoelectric point, in its dipolar zwitterionic form with a pH
of 6 is more acidic than the isoelectric pH for lysine (9.7), so lysine is in the cationic form. Aspartic
acid has an isoelectric pH of 2.8, so it is in the anionic form.
When a voltage is applied to a mixture of alanine, lysine, and aspartic acid at pH 6, alanine does
not move. Lysine moves toward the negatively charged cathode, and aspartic acid moves toward
the positively charged anode. After a period of time, the separated amino acids are recovered by
cutting the paper or scraping the bands out of the gel. If elctrophoresis is being used as an
analytical technique (to determine the amino acids present in the mixture), the paper or gel is
treated with a reagent such as ninhydrin to make the bands visible. Then the amino acids are
identified by comparing their positions with those of standards