CHEMISTR

TERM ASSESSMENT 2:
BUFFER SOLUTIONS
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 Table of contents
01 02 03
INTRODUCTION TYPES OF BUFFER pH OF BUFFERS
A buffer solution is one which maintains its SOLUTIONS An aqueous solution consisting of
pH fairly constant upon addition of small There are two types of buffers, a mixture of a weak acid and its
amounts of acid or base Acid and Basic conjugate base, or vice versa

04 05 06
Handerson- BUFFER
MECHANISM
RefersCAPACITY
to the amount of acid or
Hasselbalch How buffer operates? base a buffer can neutralize before
pH of Equation
acid buffer = pKa + the pH changes by a large amount.
([salt]/[acid])
01 Introduction
● Basically, buffer solutions are aqueous solutions consisting of a weak acid and a strong base or
vice versa.
● Or we can say that, Buffer solutions are aqueous solutions, consisting of a mixture of a weak
acid and its conjugate base or a weak base and its conjugate acid.
● HA+ H2O ↔ H3O+ + A−
● One important characteristic of buffer solutions is its ability to maintain a relatively constant
pH value as a result of addition of small acids.
● Moreover, pH of buffer solutions remains relatively constant during dilution. This is why buffer
solutions are used in a wide range of chemical applications, principally as reagents that
maintain a constant pH during fermentation processes.
● It is used to set the pH of food products, cosmetics and medicines.
02 TYPES OF BUFFER
SOLUTIONS
There are two types of buffer solutions, i.e. Acidic Buffer and Basic
Buffer.

BASIC
BUFFER
A weak base together with a salt of
same base with a strong acid. These are
called basic buffers.
Basic buffer example: Mixture of
ammonium hydroxide and ammonium
chloride.
ACIDIC
BUFFERS
A weak acid together with a salt of same
acid with a strong base. These are called
acidic buffers.
Acidic buffer example: Mixture of acetic
acid and sodium acetate (CH3COOH and
CH3COO-).
Phosphate Buffers
1. These are also called double salt buffers.
2. Besides 2 types of buffers i.e. acidic or Basic, a third appears to exist.
3. Phosphate-buffered saline is a buffer solution (pH ~ 7.4) commonly used in biological
research.
4. PBS has many uses because it is isotonic and non-toxic to most cells.
5. PBS has been shown to be an acceptable alternative to viral transport medium
regarding transport and storage of RNA viruses, such as SARS-CoV-2.
6. Some formulations do not contain potassium and magnesium, while other ones contain
calcium and/or magnesium (depending on whether or not the buffer is used on live or
fixed tissue: the latter does not require KCl or MgCl 2 ).
7. This buffer system is composed of 2 salts:
I. Monobasic potassium phosphate (KH2PO4)
II. Dibasic potassium phosphate (K2HPO4)
pH Of Buffer
Solutions
Let's illustrate the buffer action by taking example of common
buffer system consisting of solution of acetic acid and sodium
acetate.

CH3COOH ⇌ H+ +CH3COO-
CH3COONa -> Na+ + CH3COO-
Since the salt is completely ionized, it provides common ion CH3COO in excess…

The pH of Buffer changes only slightly

The pH of Buffer solution is slightly
upon addition of an acid or base.
higher than acid
Preparation of Buffer Solution
As discussed earlier, these solutions are prepared by mixing the weak bases with
their corresponding conjugate acids, or by mixing weak acids with their
corresponding conjugate bases.

If the dissociation constant of the acid (pKa) and of the base (pKb) are known, a
buffer solution can be prepared by controlling the salt-acid or the salt-base ratio.

An example of this method of preparing buffer solutions can be given by the

preparation of a phosphate buffer by mixing HPO42- and H2PO4-. The pH
maintained by this solution is 7.4.
04 Handerson-Hasselbalch Equation
Preparation of Acid Buffer
Consider an acid buffer solution, containing a weak acid (HA) and its salt (KA) with a
strong base(KOH). Weak acid HA ionizes, and the equilibrium can be written as-

HA + H2O ⇋ H+ + A−

Acid dissociation constant = Ka = [H+] [A–]/HA

Taking, negative log of RHS and LHS:

Continued…
pH of acid buffer = pKa + log ([salt]/[acid])

The equation is the Henderson-Hasselbalch equation, popularly known as the Henderson

equation.

Preparation of Base Buffer

Consider base buffer solution, containing a weak base (B) and its salt (BA) with strong acid.

pOH, can be derived as above,

pOH of a basic buffer = pKb + log ([salt]/[acid])

pH of a basic buffer = pKa – log ([salt]/[acid])

The pKa value is a quantitative information of the acid strength. It is assumed
that weak acids have pKa values greater than 3 (-log(0.001) = 3) The higher
the pKa value, the weaker the acid.

Based on the Henderson-Hasselbalch equation, it can be seen that the pH of the

buffer solution is equal to the pKa value of the acid, when the ratio of undissociated
acid concentration to the anion concentration (resulting from the dissociation of this
acid) equals 1, because log(1) = 0.

In the case of weak acid titration with a strong base, this situation occurs after
adding the amount of base equal to half of the amount needed to completely
neutralize the acid when the titration rate is 50% (Fig.). In addition, the pH of the
solution changes relatively slowly in the buffering range, ie at pH = pKa ± 1.

However, addition of too big amount of an acid or base to a buffer solution will
lead to an overload of a buffer causing a significant change in the pH of the
solution.
Example of weak acid titration curve. Titration of acetic acid solution (pKa = 4.7) with strong base (e.g.
NaOH).
 Handerson Equation
Significance of Handerson Equation
Handerson Equation can be used to:
Calculate the pH of the buffer prepared from a mixture of the salt and weak acid/base.
Calculate the pKa value.
Prepare buffer solution of needed pH.
Limitations of Henderson-Hasselbalch Equation
The Henderson – Hasselbalch equation cannot be used for strong acids and strong bases.
 05 Mechanism Of Buffer Action
The property of the solution to resist the changes in its pH value on the addition of small amounts of strong
acid or base is known as buffer action”.

In solution, the salt is completely ionized and the weak acid is partly ionized.
CH3COONa ⇌ Na+ + CH3COO–
CH3COOH ⇌ H+ + CH3COO–
On Addition of Acid and Base

1. On addition of acid, the released protons of acid will be removed by the acetate ions to form an acetic acid
molecule.
H+ + CH3COO– (from added acid) ⇌ CH3COOH (from buffer solution)

2. On addition of the base, the hydroxide released by the base will be removed by the hydrogen ions to form
water.
HO– + H+ (from added base) ⇌ H2O (from buffer solution)
06 Buffer Capacity
The number of millimoles of acid or base to be added to a litre of buffer solution to change the pH by
one unit is the Buffer capacity of the buffer.

Β = millimoles /(ΔpH)

The term "buffer capacity" (β) quantifies the change in pH of the solution caused by the addition of a
strong acid or base. It is calculated in relation to 1 dm3 of a buffer solution.
β ≈ |∆n/∆pH|
β – buffer capacity,
Δn – amount of added acid/base to the buffer solution [mol],
ΔpH – pH change cause by the addition of acid/base.
CONTINUED…
The value of the buffer capacity is strongly related to the concentrations of
ingredients used and increases with their increase. Buffer solutions with a pH
equal to the pKa value of the acid (used to make this solution) have the greatest
buffering capacity.
FORENSIC SCIENCE
AND BUFFERS
 USES IN FORENSIC
SCIENCE
The use of phosphate buffered saline for the recovery of
biological cells from swabs.

Phosphate buffered saline (PBS) is a non-toxic solution used in many

biological laboratories. Unlike water, PBS prevents cells rupturing or
shrivelling up due to osmosis. This study demonstrates that PBS can be
used for the extraction of cells from swabs and that PBS does not affect
subsequent DNA profiling.

Tissue storage solution for preservation and transfer of forensic specimen in high ambient-
temperature

Storage of tissue samples in high ambient-temperature can affect the quality of forensic evidence. Experiments
were conducted to investigate the potential use of 3 tissue storage solutions for the preservation and transfer of
forensic specimen in high ambient temperature conditions and Longmire’s buffer was found useful to store the
specimens for less than a week.
THANKS!
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