pH, Buffers and

Isotonic Solutions
pH
Definition
pH is defined as negative logarithm of
hydrogen ion concentration.

pH = - log [H3O+] OR pH = log 1/[H3O+]

pH = -log10 [H+]

Where [H+] is the concentration of H+ ion in

solution
2
 pH
Sorensens pH Scale
Aqueous solns acidic, basic, neutral contains H + and OH-
ions.
The concentration of H+ is 1 x 10-7 and for OH- is 10-7
Therefore the product is 1 x 10-14 which is a constant.
The H+ ion concentration varies from 1- 14 which is conveniently
measured by using Sorensens pH scale to express acidity
and alkalinity of the solution and they are measured by using
H+ ion concentration.
pH = 1-7 indicated acidic solution
pH = 7 indicates neutral solution
pH = >7 indicates basic solution
At pH 7 [H +] = [OH-]
pH of 1N HCl = 0 & pH of 1N NaOH = 14 3
4
Applications of pH
Enhancing Solubility
NaSalicylateprecipitates in acidic pH as Salicylic acid. Acidic
drugs are more soluble in basic pH & vice versa
Inceased Stability
Epinephrine eye drops pH 2.2 4.5
Calcium gluconate inj pH 6.0 8.2
Chloramphenicol ear drops pH 4.0 8.0
Improving Purity
Insulin
precipitates from aqueous solutions pH 5.0 6.0
Amphoteric compounds are least soluble at their isoelecric point

5
Applications of pH
Optimizing Biological Activity
Optimum pH values are required for enzyme acition. Eg Pepsin
pH 1.5

Comforting the Body

Blood pH 7.4
Saliva pH 6.4
Gastric juices pH 1.0 3.0
Instestinal juices pH 5.0 7.0

Storage of Products
Type III containers leach alkali into the formulation
resulting in increased pH. In such cases either a buffer is
employed or Type II or Type I (for parenteral preparations)
glass container have to be employed.
6
pH- Determination
pH Indicators.
pH can also be found out using universal
indicators
Colorimetric method : The color of buffers of
known pH can be matched with the colors of
unknown soution
pH can be determined using pH meter
Conductivity meter can be used to determine the
pH by measuring the conductivity of the soln

7
pH- Determination

ELECTROMETRIC METHOD
More accurate

COLORIMETRIC METHOD
Involves comparison of solutions of known pH

8
 pH Indicators
The pH indicator is defined as a solution when added
to a test solution produces a color change, which helps
in determining the pH of the test solution.
Indicators may be weak acid or a weak base
They exist as two tautomeric forms which differ in
distribution of their valency electrons.
As the pH varies, the color of the solution changes due
to changes in the degree of ionization.
Eg Methyl red pH 4.0 RED & 6.0 YELLOW COLOR

9
Principle of Color change of Indicators
Unionized benzenoid form,

Ionized Quinonoid form,

PINK COLOR
COLORLESS

Phenolphthalein
The color of the indicator depends on the ionized form and
unionized form of the indicator.
The con of the unionized form = ionized form when pH = pKa of the
indicator
Maximum sensitivity of the indicator is seen at this point (pKa
value).
And the range of the pH lies 1.5 units in either side of the pka
value.
10
Buffers & Buffering Agents
BUFFER
Buffer solutions are solutions of compounds or mixtures
of compounds which resist changes in their pH upon
addition of small quantities of an acid or alkali.
Most buffer solutions usually consist of a weak acid
and one of its salts or a weak base and one of its salts.

BUFFER ACTION
The ability of certain solutions to resist change in their
pH upon addition of an acid or a base is known as the
Buffer action

11
What is a buffer and what is not a
buffer?
Can NaCl and HCl or NaCl and NaoH
mixture be a buffer?
pH of NaCl soln = 7
Nacl Soln + 1ml of HCl , pH = 3
Nacl Soln + 1ml of NaOH, pH = 11
Therefore the above mixture is not a buffer
solution
12
Mechanism of Buffer action
Acetic acid (weak acid) and Sodium Acetate (salt of same
acid)
Acetic acid = slightly dissociated acid
Sodium acetate = completely dissociated salt
Therefore such a mixture consists of
CH3COOH molecules and CH3COO- & Na+ ions
If a strong acid is added to the above mixture, the H+ ions
supplied by the acid are immediately taken up by CH3COO-
ions to form the very slightly dissociated CH3COOH.
H+ + CH3COO- = CH3COOH
If a strong base added, the OH- ions supplied by the base are
neutralized by acetic acid present in the mixture and again
there is very little change in pH of the solution
OH- + CH3COOH = CH3COO- + H2O
13
 Mechanism of Buffer action
A mixture of a weak base and its salt also behaves in a similar
manner. E.g. equimolar soln of NH4OH & its largely
dissociated salt NH4Cl.
The mixture contains NH4OH (undissociated) molecules and
NH4+ & Cl- ions.
If a strong acid is added to this mixture, the H + ions supplied
by the acid are neutralized by the base NH 4OH.
H+ + NH4OH = H2O + NH4+
If a strong base is added, the OH- ions are neutralized by NH4+
ions forming slightly dissociated NH 4OH
OH+ + NH4+ = NH4OH
In both the cases there is very little change in the pH
14
 Buffer Equation
Henderson Hasselbalch equation

Buffer Equation helps in calculating

The pH of a buffer solution and
The change in the pH upon addition of an acid or
base.

Buffer Equation is an expression developed by

considering the effect of a salt on the ionization
of a weak acid or a weak base when the salt and
the acid or base have an ion in common.

15
 Buffer Equation for a
Weak Acid and its Salt / Common Ion Effect
Let us take an example of the effect of addition of sodium acetate
on the ionization of acetic acid. Both sodium acetate and acetic
acid have an ion common between them, i.e., CH3COO-
The dissociation constant for the acid is given by :
CH3COOH = CH3COO- + H3O+
Ka = [CH3COO-][H3O+]
[CH3COOH]
If sodium acetate is added to the acetic acid soln, it ionizes to give
acetate ion.
CH3COONa ----- CH3COO- + Na+
This causes a momentary increase in the concentration of CH3COO-
in the soln. To re-establish the dissociation constant for the
solution, the hydrogen ion term in the numerator is
instantaneously decreased. This results in the increase of the
concentration of CH3COOH in the denominator
16
 Thus the reaction,
H3O+ + CH3COO- = CH3COOH + H2O
is favoured and the Ka remains unaltered.
In other words, the ionization of acetic acid is repressed upon
the addition of the common acetate ion to the solution and
this is known as the common ion effect
The pH of the buffer soln can be obtained by rearranging the
above equation for the dissociation constant :
[H3O+] = Ka [CH3COOH]
[CH3COO-]

Since acetic acid ionizes only slightly, the conc of acetic acid
may be considered to represent the total conc of the acid in
the soln. Hence, the term [CH3COOH] = [acid]. Similarly, the
acetate ion is contributed almost entirely by the salt, NaAc,
hence, the term [CH3COO-] = [salt]
Therefore, [H3O+] = Ka [acid]
[salt] 17
Expressing in log form,
log [H3O+] = log Ka + log [acid] - log [salt]

Reversing the sign

log [H3O+] = -log Ka log [acid] + log [salt]

pH = pKa + log [salt]

[acid]
This is known as the buffer equation or the Henderson Hasselbalch
equation for a weak acid and its salt.

The buffer equation is useful expression used in the preparation of buffered

pharmaceutical solutions. It is satisfactory for calculations within the pH
range of 4 to 10.

In actual conditions, there is small differences in the pH of the soln since the
activity coefficient of the components varies with con. Taking into a/c the
activity coefficient, the H-H Eqn can be modified as:
pH = pKa + log [salt] / [acid] log AC
Where AC = activity coefficient of common ion
18
 Buffer Equation for a Weak Base and its Salt
Buffers prepared with weak bases = poor stability, therefore not
used commonly.
The buffer equation for a weak base and its salt:
[OH-] = Kb [base] / [salt]
Since the ionic product of water Kw = H3O+ X OH-
Or OH - =Kw / H3O+
On substituting the value of OH-, we get, Kw = Kb [base]
[H3O+] [salt]
Expressing in log terms,
log Kw log [H3O+] = log Kb + log [base] log [salt]
Or -log [H3O+] = -log Kw + log Kb +log [base] log [salt]
Or pH = pKw - pKb + log [base]
[salt]
19
Buffer Capacity
The Buffer capacity of a solution is a measure of its magnitude of
resistance to change in pH on addition of an acid or a base.
It is also referred to as buffer index, buffer efficiency, buffer
coefficient or buffer value.
The buffer capacity has been defined as the ratio of the increment
of a strong base (or acid) to the small change in pH brought about
by this addition.
= B
pH
Where B = small increment in gm per lt of a strong base (or acid)
added to the buffer to bring about a pH change of pH

20
According to the eqn, a soln has a buffer capacity of 1 when 1 lt
of it requires 1 gm of a strong base (or acid) to change the
pH by 1 unit.

Hence buffer capacity can also be defined as the amt of a strong

acid or strong base (expressed in gm per lt) required to
change the pH of 1 lt of a buffer system by one unit.
The smaller the pH change in a soln after addition of a
specified amt of acid or base, the greater is the buffer capacity
of the solution.
More exact eqn
= 2.303 C Ka [H3O+]
(Ka + [H3O+])2
Where C = total buffer con, i.e., the sum of the molar con of the
acid and the salt 21
A buffer solution containing a weak acid and its salts has a
maximum buffer capacity (max) when the pH is equal to the
pKa value for the weak acid or in equivalent terms
when [H3O+] equals Ka.

Substituting [H3O+] for Ka in the above eqn, we get

max = 2.303C [H3O+]2
(2 [H3O+])2

max = 2.303 C = 0.567 C

4
For most buffer systems, a total buffer con of between 0.5 and
0.05 mol per lt gives an adequate buffer capacity over a pH
range of about 2 units. 22
Factors affecting pH of Buffer solutions

Temperature Effects
Dilution Effects
Salt Effects

23
 Factors affecting pH of Buffer solutions
- Temperature Effects
The activity coefficient and the pKa value of a
buffer are dependent on the temperature.
Hence, buffer pH changes with temperature.
Buffer consisting of a base and its salt show
greater changes with temp.

An increase in temp lowers the pH of a buffer soln

containing boric acid and sod borate and raises
the pH of a buffer soln containing acetic acid and
sod acetate.
24
 Factors affecting pH of Buffer solutions -
Dilution Effects
Dilution of an aq buffer soln with water in moderate quantities
shows only a small effect on the pH of the buffer soln.
Dilution of an acidic buffer shows an increase in pH while
dilution of a basic buffer shows a decrease.

Quantification of the dilution effect can be done by the use of

dilution value which has been defined as the change in
pH brought about by the dilution of a buffer solution with
an equal volume of water .

The dilution values for most pharmaceutical buffer systems

are usually less than 0.1 pH units.

25
Factors affecting pH of Buffer solutions
Salt effects
When a salt is added to a dil buffer soln, the
acitivity coefficients of the ions are lowered.
Hence, salt added to acidic buffer lowers its pH
while that added to a basic buffer increases its
pH.
However, the change in pH is not greater than
0.1 pH units provided the final con of the neutral
salt added is not greater than that of the buffer
system itself

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 Biological Buffers
pH of blood = 7.4 by

1 buffer components in plasma

2 buffer components in erythrocytes

Plasma = 1) carbonic acid carbonate

2) acid/alkali sod salts of phosphoric acid

Erythrocytes = 1) oxyhaemoglobin haemoglobin

2) acid/alkali (K) salts of phosphoric acid

Buffer capacity = 0.025 0.039 gm per lt per pH unit.

Physiological pH range of blood = 7.0 to 7.8. below and
above which life is in danger. (acidosis & alkalosis).
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 Measurement of Tonocity

Hemolytic Method

Colligative Properties

28
ISOTONIC SOLUTIONS

The solution has essentially the same salt

concentration and hence the same osmotic
pressure as the red blood cell contents and is
said to be isotonic with blood.

29
 Hemolytic Method
RBCs are suspended in the solution whose tonocity
is to be determined. If the soln cause shrinkage of
the cells, they are said to be hypertonic and if they
cause haemolysis, they are said to be hypotonic.

Quantitative measurements are also possible using

this method based on the fact that hypotonic solns
liberates oxyhaemoglobin in direct proportion to the
number of cell haemolysed.

30
31
 Colligative methods
Colligative properties of solutions
properties that depend upon the concentration of
solute molecules or ions, but not upon the
identity of the solute.
Colligative properties include
freezing point depression,
boiling point elevation,
vapor pressure lowering, and
osmotic pressure.

32
Calculation of Tonocity using Liso Values

The Liso value for 0.9% NaCl is Tf = -0.52

C= 0.154M
Liso = Tf / C = -0.52 / 0.154 = 3.4

Tf = freezing point depression , C = molar con.

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34
METHODS OF ADJUSTING TONICITY AND pH
One of several methods can be used to calculate the quantity of sodium
chloride, dextrose, and other substances that may be added to solutions of
drugs to render them isotonic.
The methods are divided into two classes
class I methods,
sodium chloride or some other substance is added to the solution of
the drug to lower the freezing point of the solution to 0.52C and
thus make it isotonic with body fluids.
Under this class are included
The cryoscopic method and the sodium chloride equivalent
method.

35
 The class II methods,
water is added to the drug in a sufficient amount
to form an isotonic solution. The preparation is
then brought to its final volume with an isotonic or
a buffered isotonic dilution solution.
Included in this class are
the WhiteVincent method and

the Sprowls method.

36
 Cryoscopic or Freezing Point Depression
Method
The freezing point depressions of a number of drug
solutions, determined experimentally or theoretically,
are given in table

The freezing point depressions of drug solutions that

have not been determined experimentally can be
estimated from theoretical considerations, knowing
only the molecular weight of the drug and the Liso
value of the ionic class.

37
38
 Isotonicity
How much sodium chloride is required to render 100 mL of a 1% solution of
apomorphine hydrochloride isotonic with blood serum?
From Table it is found that
1% solution of the drug has a freezing point lowering of 0.08C.
1% solution of sodium chloride has a freezing point lowering of 0.58C.
To make this solution isotonic with blood, sufficient sodium chloride must be added to
reduce the freezing point by an additional 0.44C (0.52C 0.08C).

Thus, 0.76% sodium chloride will lower the freezing point the required 0.44C and will render the
solution isotonic.
The solution is prepared by dissolving 1.0 g of apomorphine hydrochloride and 0.76 g of sodium
chloride in sufficient water to make 100 mL of solution .

39
 Cryoscopic or Freezing Point
Depression Method
Blood Plasma, Lachrymal secretion FP = -0.52 C. Hence
all solns having FP = -0.52 will be isotonic solutions
% w/v of adjusting substance = 0.52 a

b
a = depression in FP due to the unadjusted solution or
substance.
b = depression in FP of 1% w/v of adjusting substance
% w/v of adjusting substance = 0.52 0.08
0.58

40
Example
How much NaCl needed to render 100ml
of 1% soln of Procaine HCl isotonic with
blood serum. A 1% soln of Procaine HCl
has a FP depression of 0.12 C

41
 Sodium Chloride Equivalent
tonicic equivalent of a drug is the amount of sodium chloride
that is equivalent to (i.e., has the same osmotic effect as) 1 g, or other
weight unit, of the drug.

E is the weight of NaCl with the same freezing point depression as 1 g of

the drug.

Calculate the approximate E value for a new amphetamine hydrochloride

derivative (molecular weight 187).
Because this drug is a uni-univalent salt, it has an Liso value of 3.4. Its E value is
calculated from equation

42
 Sodium Chloride Equivalent Method
NaCl of a drug is the amount of NaCl to 1g of drug
osmotically. It is designed as E.
This method involves simply multiplying the quantity of
the given drug by the NaCl and subtracting this value
from 0.9g of NaCl which produces an isotonic solution
with body fluids in 100ml of solution.
% NaCl = 0.9-( xE)
x = g of drug present in 100ml solution
E = Sodium Chloride equivalent

43
E values of Drugs & Added Substances

Solution, 1% w/v drug Sodium Chloride

equivalent E
NaCl 1
Apomorphine HCl 0.14
Boric acid 0.50
Calcium gluconate 0.16
KCl 0.76
Pilocarpine nitrate 0.23
44
A solution contains 1.0 g of ephedrine sulfate in a volume of
100 mL. What quantity of sodium chloride must be added
to make the solution isotonic?
The ephedrine sulfate has contributed a weight of material
osmotically equivalent to 0.23 g of sodium chloride.
% NaCl = 0.9-( xE)

= 0.9-(1X0.23)

= 0.67

0.9 g of sodium chloride is required for isotonicity, 0.67 g

(0.90 0.23 g) of NaCl must be added for 100ml sol.

45
 White-Vincent Method
This method involves the addition of sufficient qty of
water to a drug in order to prepare an isotonic solution.
An isotonic or a buffered isotonic solution is then added
to this drug solution to give the final volume.
The volume of water required for a drug to obtain an
isotonic soln can be calculated from the following
equation :
V = w x E x 111.1
V = vol in ml of an isotonic soln that can be prepared by
dissolving w gm of drug in water;
E = NaCl of the drug and
111.1 is a constant representing the vol in ml of isotonic
soln obtained by dissolving 1 g of NaCl in water
46
 Sprowls Method

Modification of White Vincent method

which uses tables listing the volume V of
isotonic solution that can be prepared by
mixing 0.3g of a drug in water

47
The weight of the drug, w,was arbitrarily fixed.
Sprowls chose as the weight of drug 0.3 g, the
quantity for 1 fluid ounce of a 1% solution.

The volume, V, of isotonic solution that can be

prepared by mixing 0.3 g of a drug with sufficient
water can be computed for drugs.

commonly used in ophthalmic and parenteral

solutions.

48