BCH 201: GENERAL
BIOCHEMISTRY 1
TITLE: pH, Buffers solutions and Titration curves
BY
Dr. Enitan O. Adesanya
pH of a solution
pH : potential of hydrogen
is a measure of how acidic/basic a substance is.
pH is effectively a measure of the concentration of hydrogen ions (that is, protons) in a substance.
The pH of an aqueous solution is based on the pH scale which typically ranges from 0 to 14 in
water.
A pH of 7 is considered to be neutral. A pH of less than 7 is considered acidic. A pH of greater
than 7 is then considered basic.
Acidic solutions have high hydronium concentrations and lower hydroxide concentrations.
Basic solutions have high hydroxide concentrations and lower hydronium concentrations.
The measurement was originally used by the Danish biochemist Søren Peter Lauritz Sørensen
(1868-1969) to represent the hydrogen ion concentration, expressed in equivalents per litre.
Measurement of pH
Figure 1: pH meter and indicator
Self-Ionization of Water
• In the self-ionization of water, the amphiprotic ability of water to act
as a proton donor and acceptor allows the formation of hydronium
(H3O+) and hydroxide ions (OH−).
• In pure water, the concentration of hydronium ions equals that of
hydroxide ions.
• At 25 oC, the concentrations of both hydronium and hydroxide ions
equal 1.0×10−7. The ion product of water, Kw, is the equilibrium
condition for the self-ionization of water and is expressed as follows:
• Kw=[H3O+][OH−]=1.0×10−14
• Sorensen defined pH as the negative of the logarithm of the
concentration of hydrogen ions.
• In terms of hydronium ion concentration, the equation to determine the
pH of an aqueous solution is:
• pH=−log[H3O+](1)
• pOH: The pOH of an aqueous solution, which is related to the pH, can
be determined by the following equation:
• pOH=−log[OH−](2)
• This equation uses the hydroxide concentration of an aqueous
solution instead of the hydronium concentration.
Relating pH and pOH
• Another equation can be used that relates the concentrations of
hydronium and hydroxide concentrations.
• Kw=[H3O+] [OH−]=1.0×10−14
• pKw=pH+pOH=14.
Ionisations of acids and Bases
• A. Strong Acids and Strong Bases
• The ionization of strong acids and strong bases in dilute aqueous solutions essentially goes
to completion.
• In aqueous solutions of strong acids and strong bases, the self-ionization of water only
occurs to a small extent.
• Since it only occurs to a small extent, the self-ionization of water is an insignificant source
of hydronium and hydroxide ions.
• Knowing this, we can say in calculating hydronium concentration in an aqueous solution of
a strong acid that the strong acid is the main source of hydronium ions.
• While in calculating hydroxide concentration in an aqueous solution of a strong base the
strong base is the main source of hydroxide ions. This is usually true unless the solutions of
strong acids and strong bases are extremely dilute.
Ionisation cont.
• B. Weak Acids and Weak Bases
• Weak acids only partially dissociate in aqueous solutions and reach a condition of equilibrium, therefore how
much they dissociate is given by the equilibrium equation for that acid in the solution:
• Ka=[H3O+][A−]/[HA](3)(3)
• with
[H3O+] is the Hydronium Concentration
[A−] is conjugate base concentration
[HA] is the Weak Acid concentration
• Weak bases also only partially dissociate in aqueous solutions and reach a condition of equilibrium. The equation
for the partial dissociation of a base is then the equilibrium equation for that base in solution:
• Kb=[OH−][B+][B](4)
• [OH−]=Hydroxide Concentration
• [B+]=Ion
• [B]=WeakBase
•
Applications of pH
In agriculture: the pH is probably the most important single property
of the moisture associated with a soil, since that indication reveals
what crops will grow readily in the soil and what adjustments must be
made to adapt it for growing any other crops.
Enzymes binding potentials: Most enzymes are proteins, they are
sensitive to changes in the hydrogen ion (H+) concentration or pH.
Buffer Solution
An acid-base buffer solution is a solution that can maintain its hydrogen ion
concentration (pH) with only minor changes when diluted or when a small
amount of acid or base is added to the solution.
• Buffer Solutions are used in fermentation, food preservatives, drug delivery,
electroplating, printing, the activity of enzymes, and the oxygen-carrying
capacity of the blood.
• They require a specific hydrogen ion concentration to function properly (pH).
• Solutions of a weak acid and its conjugate base, or a weak base and its
conjugate acid, can maintain pH and are therefore known as buffer solutions
Types of buffers solutions
• 1. Acidic Buffers
• These solutions, as the name implies, are used to keep acidic
environments intact and healthy. pH of an acid buffer is acidic, and it is
made by mixing a weak acid and its salt with a strong base to form an
acid buffer. The pH of an acetic acid and sodium acetate aqueous
solution is 4.74 when the concentrations of the two acids are equal.
• I. The pH of these solutions is lower than seven.
• II. Essentially, these solutions are composed of a weak acid and a
weak acid salt.
Types cont.
• 2. Alkaline Buffers
• These buffer solutions are used to keep the fundamental conditions in
place. It is possible to create a basic buffer by mixing a weak base and
its salt with a strong acid to produce a basic pH. The pH of an
ammonium hydroxide and ammonium chloride aqueous solution is
9.25 when the concentrations of the two acids are equal.
• Moreover, the pH of these solutions is greater than seven.
• A weak base and a weak base salt are present in these formulations.
• An alkaline buffer solution, such as a mixture of ammonium hydroxide
and ammonium chloride (pH = 9.25), is an example of a base.
Preparation of Buffer
Solution
If the dissociation constants of the acid (pKa) and the base (pKb) are known, it is possible to
prepare a buffer solution by controlling the salt-acid or salt-base ratios, respectively.
• It composes:
1. weak conjugate acid-base pair, or either a weak acid and its conjugate base or
2. weak base and its conjugate acid, must make up a buffer. When making the buffer, using one or
the other will just rely on the required pH. For example, the following could function as buffers
when together in solution:
a. Acetic acid (weak organic acid w/ formula CH3COOH) and a salt containing its conjugate base,
the acetate anion (CH3COO-), such as sodium acetate (CH3COONa)
b. Pyridine (weak base w/ formula C5H5N) and a salt containing its conjugate acid, the pyridinium
cation (C5H5NH+), such as Pyridinium Chloride.
c. Ammonia (weak base w/ formula NH3) and a salt containing its conjugate acid, the ammonium
cation, such as Ammonium Hydroxide (NH4OH)
Henderson-Hasselbalch Equation
Preparation of Acid Buffer
An acid buffer solution containing a weak acid (HA) and its salt (KA) combined with a strong
base (BB) is considered (KOH). It is possible to write down the equilibrium as:
Preparation of Base Buffer
Consider the case of a base buffer solution that contains a weak base (B) and its salt (BA) in
the presence of a strong acid.
As previously stated, pOH can be calculated.
The pOH of a basic buffer is calculated as pKb + log ([salt]/[acid]).
The pH of a basic buffer is calculated as pKa – log ([salt]/[acid]).
• Significance of Henderson-Hasselbalch Equation
• The Henderson-Hasselbalch Equation can be used to:
• 1. Calculate the pH of a buffer prepared from a mixture of the salt
and a weak acid/base; and
• 2. determine the value of pKa.
• 3. Prepare a buffer solution with the appropriate pH.
• The Henderson-Hasselbalch Equation has some limitations.
• The Henderson – Hasselbalch equation cannot be used with strong
acids or bases because of the nature of the equation.
pH Maintenance
• Consider the following example of a buffer solution containing sodium
acetate and acetic acid to better understand how buffer solutions
maintain a constant pH.
• This example illustrates how almost completely the sodium acetate
undergoes ionisation whereas only a weak amount of ionisation
occurs in the case of the acetic acid. The following is an example of an
equilibrium reaction:
What is Acid-Base Titration?
An acid-base titration is an experimental technique used to acquire
information about a solution containing an acid or base.
Hundreds of compounds both organic and inorganic can be determined
by titration based on their acidic or basic properties.
Acid is titrated with a base and base is titrated with an acid. The
endpoint is usually detected by adding an indicator.
Titration Curves
As base is added to acid at the beginning of a titration, the pH rises
very slowly. Nearer to the equivalence point, the pH begins to rapidly
increase.
If the titration is a strong acid with a strong base, the pH at the
equivalence point is equal to 7.
A bit past the equivalence point, the rate of change of the pH again
slows down.
A titration curve is a graphical representation of the pH of a
solution during a titration.
Figure 2.1 and 2.2: A titration curve shows the pH changes that occur during the
titration of an acid with a base. On the left, the base is being added to acid. On
the right, acid is added to the base. In both cases, the equivalence point is at pH
7.
Titration curve cont.
Titration curves can also be generated in the case of a weak acid-
strong base titration or a strong acid-weak base titration.
The general shape of the titration curve is the same, but the pH at the
equivalence point is different.
In a weak acid-strong base titration, the pH is greater than 7 at the
equivalence point. In a strong acid-weak base titration, the pH is less
than 7 at the equivalence point.
Summary of Titration curve
• 1. A titration curve is a graphical representation of the pH of a
solution during a titration.
• 2. In a strong acid-strong base titration, the equivalence point is
reached when the moles of acid and base are equal and the pH is 7.
• 3. In a weak acid-strong base titration, the pH is greater than 7 at
the equivalence point.
• 4. In a strong acid-weak base titration, the pH is less than 7 at the
equivalence point.
