Acidic and Basic Properties of

amino acids.

Titration of amino acids.

Henderson - Hasselbalch
equation

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• Amino acids in aqueous solution contain
weakly acidic α-carboxyl group and weakly
basic α-amino groups;
• Each of the acidic and basic amino acids
contains an ionizable group in its side chain;
• Both, free amino acids and some amino acids
combined in peptide linkages can act as
buffers.

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• Concentration of protons in aqueous solution
is expressed as pH.

• The quantitative relationship between the pH

of the solution and concentration of a weak
acid (HA) and its conjugate base (A-) is
described as Henderson - Hasselbalch
equation.

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 Henderson - Hasselbalch equation
• Describes the behavior of weak acids and
buffers;
• Describes the derivation of pH as a measure of
acidity in biological and chemical systems;
• For estimating the pH of a buffer solution and
finding the equilibrium pH in acid-base
reaction.

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• Larger Ka – stronger the acid;
• Smaller Ka – weaker the acid, the less acid has
dissociated;
• Smaller pKa – stronger acid;

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• The pKa value is one method used to indicate
the strength of an acid. 
• pKa is the negative log of the acid dissociation
constant or Ka value.
• A lower pKa value indicates a stronger acid.
That is, the lower value indicates the acid
more fully dissociates in water.

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 Buffers
• Buffer is solution that resist change in pH
following the addition of an acid or base.
• Buffer can be created by mixing a weak acid
with its conjugate base.

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• The maximum buffering capacity occurs at a
pH equal to the pKa;
• If the amount of HA and A are equal, the pH is
equal to the pKa;

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• Titration is the slow
addition of one solution of
a known concentration
(called a titrant) to a known
volume of another solution
of unknown concentration
until the reaction reaches
neutralization, which is
often indicated by a color
change;
• A titration curve is a
graphical representation of
the pH of a solution during
a titration.

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Joke

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 Titration curve of acetic acid
The maximum
buffering capacity
occurs at a pH equal
to the pKa;

If the amount of HA
and A are equal, the
pH is equal to the
pKa;

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 Dissociation of the carboxyl group

The isoelectric point (pI, pH(I), IEP), is the pH at which a molecule

carries no net electrical charge or is electrically neutral in the
statistical mean.
Zwitterion is an ion carrying both a positive and a negative charge
in different parts of the molecule.

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Figure shows the changes in pH that occurs during the
addition of base to the fully protonated form of alanyne
(I) to produce the completely deprotonated form (III).

• -COOH/-COO- pair can

serve as a buffer in the pH
region around pK1 ;
• -NH3+/-NH2 pair can buffer
in the region around pK2;
•When pH=pK1, forms I=II;
•When pH=pK2, forms II=III;

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 Isoelectric point
• pl = (2,3 + 9,1) / 2 = 5,7
• It is a midway between pK1 and pK2
• pI corresponds to the pH at which the Form II
(net charge = 0) predominates, and at which
there are also equal amounts of Forms I (net
charge of +1) and III (net charge of -1).

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 Henderson - Hasselbalch equation can be used:
• Calculate how the pH of physiologic solution
responds to changes in the concentration of a
weak acid and/or its corresponding “salt” form;

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Henderson - Hasselbalch equation can be used:

• Calculating the abundance

of ionic forms of acidic and
basic drugs.
• Most drugs are either weak
acids HA ↔ H+ + A-
• or weak bases BH+ ↔B + H+

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• Drugs passes through membrane more readily
if it is uncharged;

• Aspirin (weak acid) HA can permeate through

membranes and A- can not;
HA ↔ H+ + A-

• Morphine (weak base) B penetrates through

the cell membranes and BH+ can not;
BH+ ↔B + H+

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 The letters A through E
designate certain regions on
the titration curve for Glycine.
Which one of the following
statements concerning this
curve is correct?

A. Point A represents the region where Glycine is

deprotonated.
B. Point B represents a region of minimal buffering.
C. Point C represents the region where the net charge of
Glycine is zero.
D. Point D represents the pK of Glycine’s carboxyl group.
E. Point E represents the pI for Glycine.
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Which of this following statements concerning
the peptide shown bellow is correct?
Gly-Cys-Glu-Ser-Asp-Arg-Cys
A. The pepide contains Glutamine.
B. The peptide contains a side chain with a
secondary amino group.
C. The peptide contains a majority of amino
acids with side chains that would be
positively charged at pH 7.
D. The peptide is able to form an internal
disulfide bond.
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