PHARMACEUTICAL ANALYSIS 1

BACHELOR SCIENCE (HONS) IN

PHARMACEUTICAL TECHNOLOGY

4.3
a) Henderson–Hasselbalch
equation
b) Buffer solutions
c) Theory of Indicator

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Introduction
Buffers are solutions that maintain a relatively
constant pH when an acid or a base is added.

They therefore protect, or “buffer,” other molecules in

solution from the effects of the added acid or base.

Buffers contain either a weak acid (HA) and its

conjugate base (A−) or a weak base (B) and its
conjugate acid (BH+), and they are critically important
for the proper functioning of biological systems and
pharmaceutical preparation.

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 Unbuffered solution

A solution that is buffered

by acetic acid/acetate
Question

Which combination(s) make a simple buffer solution?

A. HCl and KCl
B. H2CO3 and NaHCO3
C. H3PO4 and NaCl
D. CH3COOH and CH3COOK

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How is the buffer works?
(Acetic acid with its conjugate base
– acetate ion)

http://www.ehu.eus/biomoleculas/buffers/mm/buffer.swf

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 Characteristics of Buffered solutions

 Contain relatively large concentrations of a weak

acid and its conjugate base.
 When acid is added, it reacts with the conjugate
base
 When base is added, it reacts with the acid.
 pH is determined by the ratio of the base and
acid.
Importance of buffer solutions
Buffer solutions are used in preparation of:
a) Dosage forms such as injections and
ophthalmic solutions which are given
directly to pH sensitive body fluids
b) In pharmaceutical analysis where pH
adjustment are required.

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Buffer equation:
The Henderson-Hasselbalch
Equation
 The change in pH upon the addition of an acid
or base and the pH of a buffer solution is
calculated by buffer equation:
 For weak acid:

pH = pKa + log [salt] / [acid]

 For weak base:

pH = pKa + log [base] / [salt]

Theory of Indicator

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Indicators
 Indicators are the compounds which give
visible colour change at the completion of
the reaction.
 Indicators are weak acid / weak base with
pKa. The colour changes of indicators is in
range of +1 and -1 of its pKa.
 Neutralization indicators or acid-base
indicators are substance which indicates
the equivalence points in neutralization.
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Theory of indicator
 The theory suggest that the colour of organic
compounds is due to presence of some
unsaturated chromophores with C=O, C=C,
N=O and N=N.
 There are three theories of indicators:
a) Litmus theory
b) Ostwald theory
c) Quinonoid theory

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 1) Litmus theory
 Indicators behave as litmus.
 Litmus is a weak acid which give blue colour in ionic
form and red colour in unionized form.
 Addition of hydroxide ion in litmus solution causes
reaction between hydroxide and hydrogen ion of litmus
releasing ionized litmus ion which give blue colour.
 Addition of hydrogen ion give excess hydrogen ion in
the system which favours the formation of litmus
(unionized form) leads to development of red colour.

Red colour Blue colour (ionized form) 12

 1) Litmus theory
Litmus paper produced from Lichens

Red colour Blue colour (ionized form) 13

2) Quinonoid theory
 According to this theory the two colour forms
of indicators are interconvertible (there are
tautomers).
 One form is benzonoid form having light color
or colourless. Another form is quinonoid form
which has deep colour.

(Colourless) (Coloured)
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Quinonoid theory

 At pH between 8.4 – 10.4, Phenolphthalein

undergoes a structural rearrangement as a
proton is removed from one of its phenol group
as the pH rises.

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3) Ostwald Theory
 According to this theory, acid base
indicator is a weak organic acid or base
which ionizes in aqueous solution to give
different colour.
 Unionized and ionized forms of indicator
have different colours which indicated the
end point.

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Ostwald Theory
- Methyl orange

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Natural Indicators

An Indicator Derived from Red Cabbage:

pH increases

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Structure arrangement of pigment
colour in red cabbage

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 Test
Theory of Indicator Definition and example

Litmus theory

Quinonoid theory

Ostwald Theory

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Strong acid/strong base titration
Strong acid/ strong base titration are used
in pharmacopoeial assays :
1) Perchloric acid
2) Hydrochloric acid
3) Sulphuric acid
4) Thiamine hydrochloride

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Choice of acid base indicator
 For acid/base donate/accept 1 proton,
there will only have 1 inflection of
titration curve
 Choice of acid base indicator will fall
within the inflection of titration curve

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 Choice of acid base indicator (1)

When 1M NaOH is
added to 25mL of 1M
solution of weak acid
aspirin (pka 3.5).

From the inflection of

titration curve,
Phenolphthalein is
suitable as indicator.

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 Choice of acid base indicator (2)
When 1M HCl is
added to 25mL of 1M
solution of weak
base quinine (pka
8.5).

From the inflection of

titration curve,
Methyl orange is
suitable as indicator.

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Self-test
Determine which of these indicators could be
used in the titration of aspirin and which
could be used in the titration of quinine?

Indicator Titration of Aspirin /

quinine
Bromphenol blue (pKa 4.0)
Methyl red (pKa 5.1)
Cresol red (pKa 8.3) .
Chlorophenol blue (pka 6.0) .

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Choice of acid base indicator (3)
 For acid/base donate/accept more than
1 proton
 If pKa values of any acidic or basic group
differ for more than 4
 there will have 2 inflection of titration
curve.

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 Choice of acid base indicator (4)
When 1M HCl is added to
25mL of 1M solution of
sodium carbonate with
(pka 10.32 and 6.38), will
have 2 titration curve.

From the inflection of

titration curve,
Phenolphthalein and
Methyl orange is suitable
as indicator.

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Self-test

How many inflections do the following substances

have in their titration curve?

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30
1. The pKa of histamine is 5.74. What is the ratio
of ionized to unionized histamine (a) at pH 5.74
(b) at pH 7.4
2. Find the pH of 1.25 M acetic acid and 0.75 M
potassium acetate, pKa = 3.5 (A: 4.0)
3.0.788 M lactic acid and 1.27 M calcium lactate.
pKa = 3.86 (A: 4.06)

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4. 0.590 M ammonium hydroxide and 1.57 M
ammonium chloride. pKa = 9.25 (A: 8.8)

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5.All of the following are true of buffers EXCEPT:

 Consist of a strong acid and its conjugate base

 Work best within one pH unit of the pKa
 Effectiveness is relative to the pH of solution and
concentration of buffer
 Buffered solutions resist changes in pH when acid
or base is added to solution

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6) Calculate the ratio of acetate/acetic acid in the
medium of pH 3 (pKa 4.74) (Ans: 0.018/1)
7) Calculate the % ionization of Acetylsalicylic acid
if acetylsalicylic dissolve in a pH 3.00, pKa=3.5
solution (Ans: 24%)

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