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
 Sorensen’s 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 Sorensen’s 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
 NaSalicylate precipitates 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
pH- Determination by
Potetiometric Method
 The [H+] con of a solution can be determined
electrometrically by measuring the potential differences
set up between an electrode immersed in the solution
and a standard electrode.

 Types of Electrodes
 Hydrogen electrode (platonized Pt in equilibrium
with hydrogen gas) (not for routine works)
 Glass electrode. (modern commercial pH meters
employ the glass electrode)

11
 Glass Electrode
 Very useful electrode for the determination
of pH of a solution

 It involves no electron exchange but allows

transfer of H+ ions through its glass membrane

 It consists of a very thin bulb made from glass

membrane having high electrical conductivity

 This bulb contains HCl of definite con and/or

Ag/AgCl wire to make electrical contact with it
12
 Glass Electrode
1. a sensing part of electrode, a bulb made from
a specific glass
2. sometimes the electrode contains a small
amount of AgCl precipitate inside the glass
electrode
3. internal solution, usually 0.1M HCl for pH
electrodes
4. internal electrode, usually silver chloride
electrode or calomel electrode
5. body of electrode, made from non-conductive
glass or plastics.
6. reference electrode, usually the same type as 4
7. junction with studied solution, usually made
from ceramics or capillary with asbestos or
quartz fiber.

13
Bulb construction & sensitivity
 The thin membrane of bulb is prepared from special type
of glass sensitive and permeable to H+ ions. Generally
the glass membrane is made from soft soda-lime glass
containing lithium silicate with lanthanum and barium
ions added to it

 The sensitivity of membrane to protons and other cations

generally depend upon the composition of glass
membrane. It is specific to H+ ions upto pH 9.0 and
above which somewhat responsive to Na+ and other
cations.

14
Principle
 The surface of the glass membrane must be hydrated before
it can function as pH electrode. The hydration of pH sensitive
glass membrane involves an ion-exchange reaction between
strongly charged cation in glass lattice and proton from the
solution.
H+ + Na+ (Cl)- = Na+ + H+ (Cl)-
soln glass soln glass
 These ions act as lattice adjusters. Thus bulbs function like a
semi permeable membrane permeable to H+ ions. The H+
ions enter glass lattice.
 It is known that on passing of electric current through the
glass membrane, the amt of H+ ions transferred through glass
nearly in accordance with Faraday’s Law. For this, presence
of water in the glass is essential.
 The pH function is impaired if the glass gets dried.
 To carry out pH measurement of a solution, glass electrode is
dipped into it and connected to a suitable reference electrode
like saturated calomel electrode and emf, recorded. 15
Principle
 Each glass electrode have its own characteristic
potential. This depends on several factors like
composition of glass, thickness of glass bulb,
size and area of glass membrane.

 The potential of glass electrode when immersed

in a solution of which pH is to be measured is
given by,
E = K – 0.0591 pH

16
pH- Determination by
Potetiometric Method
 The glass electrodes, immersed in the solution
to be investigated form half of an electrical cell,
the other half of which is a standard half – cell
such as calomel electrode.

It is the e.m.f. of this cell which is actually

measured, by means of potentiometer and the
pH is calculated from the relation:
E = K – 0.0591 pH (at 25°C)

17
Advantages
 Reasonably rapid response over a wide
pH range
 Uninfluenced by presence of oxidizing or
reducing agents.
 Viscous, Colored solns, suspension or
colloidal solns can be used.

18
Disadvantages
 Fragile
 Unsatisfactory results above pH 10
 Glass electrode must not be allowed to go
dry
 Requires conditioning for some time by
soaking in water or in a dil acid buffer soln.

19
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

20
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

21
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
22
 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 NH4OH.
H+ + NH4OH = H2O + NH4+
If a strong base is added, the OH- ions are neutralized by NH4+
ions forming slightly dissociated NH4OH
OH+ + NH4+ = NH4OH
In both the cases there is very little change in the pH
23
 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.

24
 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
25
 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 con of acetic acid
may be considered to represent the total con 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]
26
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 γAC
Where γAC = activity coefficient of common ion

27
 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]
28
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 β 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.
β = ΔB
Δ pH
Where ΔB = small increment in gm ≡ per lt of a strong base
(or acid) added to the buffer to bring about a pH change
of ΔpH
29
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
β = 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
30
A buffer solution containing a weak acid and its salts has a
maximum buffer capacity (βmax) 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

βmax = 2.303C [H3O+]2
(2 [H3O+])2

βmax = 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.
31
Factors affecting pH of Buffer
solutions

 Temperature Effects
 Dilution Effects
 Salt Effects

32
 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.
33
 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.
34
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

35
 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).
36
 Measurement of Tonocity

 Hemolytic Method

 Colligative Properties

37
 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.

38
 Colligative methods
 Calculation of Tonocity using Liso Values
 Lowering of volatile point
 Elevation of boiling point
 Depression of freezing point
 Osmotic pressure of solution

The colligative properties depends on the # of

paricles and not on the nature of solvent
The Liso value for 0.9% NaCl is
Liso = ΔTf / C = -0.52 / 0.154 = 3.4
ΔTf = freezing point depression , C = molar
con.
39
 Methods of Adjusting Tonocity

 NaCl, Dextrose, Water + Drug in con ≈ isotonic soln

 Cryoscopic or Freezing Point Depression method
 Sodium Chloride ≡ Method
 White – Vincent Method
 Sprowls Method

40
 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

41
Example
 0.76% w/v is the amount of 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

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
 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
45
 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

46
47
Faraday's 2nd Law of Electrolysis

 The number of Faradays of electric charge

required to discharge one mole of
substance at an electrode is equal to the
number of "excess" elementary charges
on that ion.

48