CHM131:

Biochemistry

LE C T U R E 7
D R . P A S H A G H A Z AL
ASSI STANT PRO FESSO R
BIOSCIENCE DEPARTMENT
Review of pH, acids, and bases
• pH is generally defined as the negative logarithm of the hydrogen ion activities
(concentration) expressed over 14 orders of magnitude

pH = -log10 [H+]

• The pH scale is a reciprocal relationship between [H+] and [OH-]

• Because the pH scale is based on negative logarithms, low pH values represent the
highest [H+] and thus the lowest [OH-]
• At neutrality, pH 7, [H+] = [OH-]
Review of pH, acids, and bases
• Strong electrolytes dissociate completely in water
• Electrolytes are substances capable of generating ions in solution
• Increase the electrical conductivity of the solution
• The dissociation of a strong acid in water

The equilibrium constant is

HCl + H2O H3O+ + Cl-

[H3O+][Cl-]
K=
[H2O][HCl]
[H2O] is constant in dilute aq. Solutions and is incorporated into the equilibrium constant. giving
rise to a new term Ka-the acid dissociation constant = K[H2O], [H3O+] is expressed as [H+]

[H+][Cl-] Because Ka is large for HCl,

Ka = [H+] in solution = [HCl] added to solution.
[HCl]
Thus, a 1M HCl solution has a pH of 0, a
1 mM HCl solution has a pH of 3, and so
on. Conversely, 0.1 M NaOH solution has
a pH of 13.
 For a strong acid Ka will approach be large because the
[H+][Cl-]
Ka= nearly all of the protons will be dissociated.
[HCl] The [H+] at equilibrium is equal to the initial concentration
of the acid.

Calculate the pH of a 1M HCl solution

HCl + H2O H3O+ + Cl-

0.0004% at equilibrium 99.996% at equilibrium

Since we are at equilibrium, H3O+ is equal to the initial concentration of acid.

[H+] = [H3O+] = [HCl] = 1M

We know that pH is the -log of [H+], therefore for 1M HCl at equilibrium

pH = -log10 [H+]
pH = -log10 (1)
pH = 0
pH [H+] [OH-]
0 (100) 1.0 0.00000000000001 (10-14)
1 (10-1) 0.1 0.0000000000001 (10-13)
2 (10-2) 0.01 0.000000000001 (10-12)
3 (10-3) 0.001 0.00000000001 (10-11)
4 (10-4) 0.0001 0.0000000001 (10-10)
5 (10-5) 0.00001 0.000000001 (10-9)
6 (10-6) 0.000001 0.00000001 (10-8)
7 (10-7) 0.0000001 0.0000001 (10-7)
8 (10-8) 0.00000001 0.000001 (10-6)
9 (10-9) 0.000000001 0.00001 (10-5)
10 (10-10) 0.0000000001 0.0001 (10-4)
11 (10-11) 0.00000000001 0.001 (10-3)
12 (10-12) 0.000000000001 0.01 (10-2)
13 (10-13) 0.0000000000001 0.1 (10-1)
14 (10-14) 0.00000000000001 1.0 (100 )
[H+] pH

10-7 = 7

10-3 = 3 pH = -Log[H+]
10-2 = 2

10-10 = 10

5x10-4 = 3.3

7x10-6 = 5.15

3.3x10-8 = 7.48
Review of pH, acids, and bases
• Weak electrolytes only slightly dissociate in water
• Acetic acid, CH3COOH
• The dissociation of a weak acid in water

CH3COOH + H2O H3O+ + CH3COO-

The acid dissociation constant is

[H+][CH3COO-]
Ka = = 1.74 X 10-5 M
[CH3COOH]

Ka is also called the ionization constant, because Ka is small, most of the acetic acid is not ionized.
 Acid dissociation constant
• The general ionization of an acid is as follows:
HA H+ + A-
So the acid dissociation constant is as follows:
[H+][A-]
Ka =
[HA]

There are many orders of magnitude spanned by Ka values, so pKa is used instead:

pKa = - log10 Ka

The larger the value of the pKa, the smaller the extent of dissociation.
pKa <2 is a strong acid
Henderson-Hasselbalch Equation
• Describes the dissociation of a weak acid in the presence of its conjugate base
• The general ionization of a weak acid is as follows:

HA H+ + A-
So the acid dissociation constant is as follows:

[H+][A-]
Ka =
[HA]
Rearranging this expression in terms of the parameter of interest [H+] gives the following:

[H+] =
Ka [HA]
[A-]
Henderson-Hasselbalch Equation
Take the log of both sides:

[HA]
log[H+] = log Ka + log
[A-]
Change the signs and define pKa as -log Ka :

[HA]
pH = pKa - log
[A-]
or
[A-]
pH = pKa + log
[HA]
 Observation

If you add .01 ml i.e 1/100 ml of 1M HCl to 1000 ml

of water, the pH of the water drops from 7 to 5!!

i.e 100 fold increase in H+ concentration: Log = 2

change.

Problem:
Biological properties change with small changes in
pH, usually less than 1 pH unit.

How does a system prevent fluctuations in pH?

 Buffers

A buffer can resist pH changes if the pH is at or near a weak

acid pK value.

Buffer range: the pH range where maximum resistance to pH

change occurs when adding acid or base. It is = +1 pH from
the weak acid pK

If pK is 4.8 the buffering range is 3.8 5.8

Why?
Henderson - Hasselbalch equation
+ -
From [ H ][ A ]
K=
[ HA]

Rearrange + [ HA]
[H ] = K -
[A ]
Take (-)Log of each [ A- ]
pH = - log K + log
[ HA]
-
[A ]
pH = pK + log
[ HA]
 -
[A ]
pH = pK + log
[HA]
-
[A ] 1 10
ratio varies from Þ
[HA] 10 1

Above and below this range there is insufficient

amount of conjugate acid or base to combine with
the base or acid to prevent the change in pH.
 For weak acids

HA A- + H+
This equilibrium depends on concentrations of each component

If [HA] = [A-] or 1/2 dissociated

-
[A ]
Then log = log 1 = 0 : pH = pK
[ HA]
By definition the pK is the pH where [HA] = [A-] :
50% dissociated
 The buffer effect can be seen in a titration curve.

To a weak acid salt, CH3C00-, add HC1 while monitoring pH vs. the
number of equivalents of acid added.
or
do the opposite with base.

Buffer capacity: the molar amount of acid which the buffer can
handle without significant changes in pH.

i.e

1 liter of a .01 M buffer can not buffer 1 liter of a 1 M solution of

HCl
but
1 liter of a 1 M buffer can buffer 1 liter of a .01 M solution of HCl
Table 2-3
Dissociation constants and pK’s of Acids & buffers

Acid K pK

Oxalic 5.37x10-2 1.27

H3PO4 7.08x10-3 2.15
Succinic Acid 6.17x10-5 4.21 (pK1)
Succinate 2.29x10-6 5.65 (pK2)
H2PO4- 1.51x10-7 6.82
NH4+ 5.62x10-10 9.25
Glycine 1.66x10-10 9.78
 Exams Problems
You will be asked to solve Henderson -
Hasselbalch type problems:

-
[A ]
pH = pK + log
[HA]

You may be asked the pH, pK, the ratio of acid or base or solve for
the final concentrations of each.
 The 6 step approach
1. Write the Henderson + Hasselbalch equation.

2. Write the acid base equation

3. Make sure either an H+ or OH- is in the equation.

3. Find out what you are solving for

4. Write down all given values.

5. Set up equilibrium conditions.

6. Plug in H + H equation and solve.

 What is the pH of a solution of that contains 0.1M CH3C00-
and 0.9 M CH3C00H?

1) pH = pK + Log [A-]
[HA]

2) CH3C00H CH3C00- + H+

3) Find pH

4) pK = 4.76 A- = 0.1 M HA = 0.9 M

5) Already at equilibrium

6) X = 4.76 + Log 0.1

0.9

Log 0.111 = -.95 X = 4.76 + (-.95) X = 3.81

What would the concentration of CH3C00- be at pH 5.3
if 0.1M CH3C00H was adjusted to that pH

1) pH = pK + Log [A-]
[HA]
2) CH3C00H CH3C00- + H+
3) Find equilibrium value of [A-] i.e [CH3C00-]
4) pH = 5.3; pK = 4.76
5) Let X = amount of CH3C00H dissociated at equilibrium

[A-] = [X]
[HA] = [0.1 - X]

6) 5.3 = 4.76 + Log [X]

[0.1 - X]

Now solve.
Numerical problem
• Calculate the pH of a buffer that contains 0.7 M
ammonia and 0.9 M ammonium chloride. (pKa =
9.248).
Solution
• By using the Henderson Hasselbalch Equation is:
pH = pK + Log [A-]
[HA]
• pH = 9.248 + log [0.9 / 0.7]
• pH = 9.248 + log 1.29
• pH = 9.248 + 0.11
• pH = 9.358