UNIT-V

pH, BUFFER and

ISOTONICSOLUTION
|Points to be covered in this topic
1. SORENSEN'S pH SCALE

+2. pH DETERMINATION (ELECTROMETRIC

& CALORIMETRIC)
3. APPLICATION OF BUFFER

+4. BUFFER EQUATION

5. BUFFER CAPACITY

6.BUFFER IN PHARMACEUTICAL AND

BIOLOGICAL SYSTEM

7. BURFER ISOTONIC SOLUTION

 OsORENSEN'SpH SCALE
pH refers topotential of hydrogen ions concentration.
Sorenson's has defined pH ofasolution asthe logarithm of the reciprocal
of thehydrogen ions or hydronium ions concentration [H,0 *]
> Mathematically
pH= log 1/ [H,0*] --. --(1)
The above equation can be rearranged as
pH=log 1-log [H,0*]------ ---(2)
=> pH=-log [H,0+]or pH=-log [H]{as the value of log 1 is Zero} --- (3)
Hence pH can also defined as the negative logarithm of hydrogen ion or
hydronium ions concentration.
The concentration of [H,0]isexpressed in molarity, mol/L
" In pure water[H]=1.0X107
" SopH of neutral (pure) water is -log (10) =7
Acidic solution:
" The solutions having [H*] value greater than 107
are called acidic solution. PH7 PH4

The solutions having [H]value less than 10 are

called basic solution.

" Hence pH value of all acidic solutions are less than 7 and pH value of all
basic solutions are greater than 7.
" Sorenson developed a scale based on the pH value and different
concentration of H,0* in asolution which is called Sorenson's pH scale
OR 'Hydrogen ion exponent.
Sorenson's scale assigns a pH of 0 to 14, with 0 being the most acidic, 14
beingthe most basic, and 7 being neutral (neither acidic nor basic).
 [H*] pH Color [OH-] pOH
100 10-14 14 ACIDIC
10-1 1 10-13 13
10-2 2 10-12 12
10-3 3 10-11 11
10-4 4 10-10 10
10-5 5 10-9 9
10-6 6 10-8
10-7 7 10-7 7
10-8 10-6
NEUTRAL
8 6
10-9 9 10-5 5
10-10 10 10-4 4
10-11 11 10-3 3
10-12 12 10-2 2
10-13 13 10-1 1 BASIC
10-14 14 100 0

Applications
> The pH of the solutions must be controlled in pharmacy particularly in
formulations of eye drops,ear drops, injections and liquid orals for the
following reasons :
The pH of the pharmaceutical preparations should
Enhancing solubility be adjusted so as to make the API soluble and
and stability
remain physically stable in the formulation.
The purity of the protein can be determined as the
Improving purity amphotericcompounds are least soluble at their
isoelectric points.
Thedrug molecules are absorbed differently from
Absorption of drugs various parts of the GIT as the later differs in their
pH.
Special type of glass is used in case the glass
Storage of products container imparts alkalinity and alters the pH of
the contents.
The pH of the formulations that are administered to
Comforting the body different tissues of the body should be optimum to
avoid irritation (eyes), haemolysis (blood) or
burning sensation (abraded surface).
Optimizing biological Enzymes have maximum activity at a definite pH
activity value.
 OpH DETERMINATION
The are two widely accepted methods for the determination of the
pH of a solution
(a) Colorimetric method
(b) Electrometric method
(a) Colorimetricmethod
Principle
" The colour comparison of the test solution to
that of the standard both treated with universal
indicator.
This method is used to determine the pH of the
solution in the pH range of 3to 11 ± 0.2 units.
Indicator strips of filter papers are used for identifying the pH.
several standard solutions can be prepared or procured which are mixed
solution of buffer and indicator.

Also Capillators and Comparators are commercially available for this

purpose.

Standard solutions (mixture of buffer solution and

universal indicator) of large volume placed in capillary
Comparators tubes are called comparators.
This is useful in examining turbid and coloured solutions.
Standard solutions (mixture of buffer solution and
Capillators universal indicator) of small volume placed in capillary
tubes are called capillators.
Method:
Step-1: Standard buffer solutions of known pH ranging from 3.0 to 11.0
are prepared with 1.0 pH interval.
 Sten-2: A few drops of universal indicator solutions are added to the
above solution that produce different colours.
Sten-3: The colour of the test solution is compared with the colour of the
standard solutions. The pH of the standard solution that has nearly same
colour as that of test is consider as the approximate pH of test solution.
Step-4: In a similar way the test solution isagain compared with the colour
of the indicator treated standard solution of narrow pH range with 0.2
pH interval.
Step-5: Step 2 and 3are again repeated and the pH of the solution is
reported.
Advantages
Less expensive
Easy estimation of pH unless the drug shows buffer action.
* Disadvantage
V'oter
" This method is less accurate and less convenient
" It is not useful for coloured or turbid solution. esarg Eetle
Teaperate

This isnot useful in presence of salts, proteins etc.

soaton

b)Electrometricmethod Clas Mrahrae

Cerde

(bre
* Principle
The magnitude in the potential difference between glass and a solution
containing hydrogen ion varies with concentration of H

concentration.

Hence the pH of the solutions are determined by means of the electrodes.

Hydrogen electrode and glass electrodes are used.
The instrument used to determine the pH of unknown solution by this
method iscalled pH meter.
 Method:
The glass electrode is attached to the instrument.
ApH meter with its control knobs.
Sten-1: At first the instrument temperature is set to that of the solution
temperature.
Sten-2: The electrode is immersed into a standard buffer solution of pH
7.0. The potential control knob is adjusted till the pH reading in digital
meter becomes 7.0.

Sten-3: Then the instrument is calibrated using standard buffersof pH

4.0(M/20 potassium hydrogen phthalate) and pH 9.14.
º Step-4:The electrode is now rinsed with distilled water properly and re
immersed into the test solution. The pH value is obtained from the
digital meter.
" The pH of the test solution can be changed by the addition of slight
amount acid or base solution.
The procedure is followed till the desired pH is obtained.
* Advantages:
It gives an accurate measurementof pH.
Glass electrode is not affected by oxidation-reduction system.
Theelectrode establishes equilibrium rapidly.
The indicator need not required.
14
* Disadvantages: 3

ALKALINE
The cost of pH meter is high compared
to colorimetric method. 0
pH
Thismethod is not suitable for viscous
solutions and gels because of poor
ionic mobility.
 INDICATORS
> The pH indicators
The pH indicator is aweak acid or weak base
that exists in tautomeric form that readily
interconvert.
It is asolution when added to test solution
produces a colour change, which helps in
determining the pH of the test solution.
The colour of any indicator depends on the pH
of the solution
Ex- Phenolpthalein, methyl red,Thymol
blue

Universalindicator
Universal indicator is defined as a mixture of AnehtMed Rogon

several indicators, which gives different Unlvreat indloator

Solution p H 4 2
color shades as the pH of the solution varies,
in aparticular pH range.
Name of Colour Universal
indicator pH range change indicator

Methyl yellow 3.1-4.4 Blue yellow

Methyl red 4.2-6.2 Red-yellow

Mixture of all

Bromothymol 6.0-7.6 Yellow- blue

blue indicator range of

Thymol blue 8.0-9.6 Yellow-blue

pH is1 to 11

Phenolpthalein 8.3-10.0 Colourless-pink

 OBUFFER
" Buffers are defined as a compound or a mixture of compounds that
resists the pH upon the addition of small quantities of acid or alkali.
Buffer have definite pH value.
Buffer action:
The resistance to a change in pH is known as buffer action.
Buffer capacity:
The amount of acid/base required to produce a unit change in pH in a
solution is called buffer capacity.
O APPLICATION OF BUFFER

The pHof the pharmaceutical formulations are adjusted

Solubility
to an optimum value so that the drug remain solubilized
enhancement
though out its shelf-life and not precipitated out.
The purity of proteins can be identified from its solubility
Improving
at their isoelectric point as they are least soluble at this
purity
point.

Optimising Enzymes have maximum activity at definite pH values.

biological Hence buffer of desired pH is added to the preparation.
activity
The pH of the formulations that are administered to
Comforting the different tissues of the body should be optimum to avoid
body irritation (eyes), haemolysis (blood) or burning
sensation (abraded surface).

Increasing To prevent hydrolysis and for maximum stability, the pH

stability of the medium should be adjusted suitably.
 Pharmacy utilizes buffer to ensure
In pharmaceutical system maximum stability of product by
adjusting their pH Level
It inhibits gastric acid production by acting as a buffer by reducing the
stomach acid content.

Buffer equation-Henderson-Hasselbalch equation

"Common lon Effect and the Buffer Equation for a Weak Acid and Its Salt"
" The pH of a buffer solution and the change in pH uponthe addition of
an acid or base can be calculated by use of the buffer equation.
Two separate equations are obtained for each type of buffer, acidic and
basic.

Buffer equation is developed by considering the effect of a salt on the

ionization of a weak acid when the salt and the acid have an ion in
common.

The buffer equation is also knowm as Henderson-Hasselbalch equation.

Acombination of a weak acid and its conjugate base or a weak base
and its conjugate acid acts as a buffer.
If 1 mL of a 0.1 N HCl solution is added to 100 mL of pure water, the
pH is reduced from 7 to3.
If the strong acid is added to a0.01 M solution containing equal
quantities of acetic acid and sodium acetate, the pH is changed only
0.09 pH units because the base Ac ties up the hydrogen ions according
tothe reaction
Ac+H,0**HAc+H,0 (1)
If a strong base, sodium hydroxide, is added to the buffer mixture, acetic
acid neutralizes the hydroxyl ions as follows:
HAC + 0H H,0+AC: (2)
For exanple:
When sodium acetate is added to acetic acid, the dissociation constant
for the weak acid, is momentarily disturbed because the acetate ion
supplied by thesalt increases the [Ac]term in the numerator.
The constant k, at 1.75 x10-5, the hydrogen ion term in the numerator
[H,0* ]isinstantaneously decreased, with a corresponding increase in
[HAC].
Therefore, the constant k, remains unaltered.
The equilibrium isshifted in the direction of the reactants.
The ionization of acetic acid is repressed upon the addition of the
common ion, Ac.
This is an example of the common ion effect.

K, = [H,0][Ac -(3)
[HAC]
ThepH of the finalsolution is obtained by rearranging the equilibrium
expression for acetic acid:

HAc+ H,0 H,0*+Ac (4)

If the acid is weakand ionizes only slightly
Expression [HAc] =Total concentration of acid.

[H,0*] =
K,[HAC] (5)
[Ac]
The slighthy ionized acidic solution, the acetate concentration [Ac ] can
be considered as having come entirely from the salt,sodium acetate.
Because 1mole of sodium acetate yields 1 mole of acetate ion,[Ac ]is
equal to the total salt concentration and is replaced by the term (Salt).
Hence, equation (5) is written as
 K,[acid] (6)
[H,0*] =
[Salt]
Equation (8-6) can be expressed in logarithmic form, with the signs
reversed, as
-log[H,0*] =-log k,-log [acid]+ log [salt] (7)
from which is obtained an equation ,known as the buffer equation or the
Henderson-Hasselbalch equation, for a weak acid andits salt:

pH. pk," log |Salt]

Acid) (8)

The ratio [Acid]/[Salt] in equation (6) has been inverted by logarithmic

operations in equation (7), and it appears in equation (8) as

[Salt]/[Acid].
pk,,the negative logarithm of K,, iscalled the dissociation exponent.
The buffer equation is important in the preparation of buffered
pharmaceutical solutions it is satisfactory for calculations within the pH
range of 4 to 10.
The Buffer Equation for a Weak Base and Its Salt
Buffer solutions are not ordinarily prepared from weak bases and their
salts because of the volatility and instability of the bases and because of
the dependence of their pH on pK,which is often affected by temperature
changes.
The buffer equation for solutions of weak bases and the corresponding salts
be derived in a manner analogous to that for the weak acid buffers.
Accordingly, [basel
[OH|=K, |salt] (9)

Using the relationship [OH] =k«/[H,0 ], the buffer equation is obtained

 [basel
pH.pK - pk, t log Salt) (10)
* Applications:
For a definite pH solution,it is essential to add salt and acid (or base)
water in a desired ratio. This ratio is determined by Henderson
Hasselbalch equation.
Since salt and acid are added in the preparation of abuffer solution,
their concentrations are known. Using this data, the resultant pH of a
solution can be calculated using buffer equation.
The pKa of various drugs can be determined fronm pH of solutions
The solubility of a substance at any pH can be predicted provided
intrinsic solubility and pK, are known.
A suitablesalt forming substance can be selected based on Henderson
Hasselbalch equation.
OBUFFER CAPACITY
Buffer counteracts the change in pH of a solution upon the addition of a
strong acid, astrong base,or other agents that tend to alter the hydrogen
ion concentration.

The magnitude of the resistance of a buffer to pH changes is referred to as

the buffer capacity (B).
" It is also known as buffer efficiency, buffer index, and buffer value.
Koppel and Spiro and Van Slyke introduced the concept of buffer
capacity.
Buffer capacity, B, is mathematically expressed as:
AB 1
ApH
 B= concentration of base (or acid) added, gram-Eq/L
Delta, Ahas its usual meaning, a finite change
AB is the small increment in gram equivalents (g.Eq)/liter of
strong base
pH change of ApH.
According to equation (1), the buffer capacity has a value of 1 when I gram
equivalent of strong base (or acid) is added to 1litre of buffer solution, if
the change in pH is I unit.
The buffer has its greatest capacity, when [salt]/[acid] is equal to 1.
Therefore, Henderson Hasselbalch equation may be written as pH = pk,.
Buffer capacity decreases appreciably as the pH deviates more than 1
unit on each side of the pK, value.
Buffer capacity is not a fixed value for a given buffer system, but depends
on the amount of base added.

Buffer capacity changes as the ratioof log [salt]/[acid] increases with

added base.

Buffer equation can be used to calculate the pH of the solution after the
addition of base.

100
80

40

20

4 5 7 10
pH
1. Approximate Calculation ofBuffer Capacity
Acetate buffer containing 0.1 mole each of acetic acid and sodium
acetate in 1 liter of solution.

To this are added 0.01-mole portions of sodium hydroxide.

When the first increment of sodium hydroxide is added, the concentration
of sodium acetate, the [salt] term in the buffer equation, increases by
0.01 mole/liter and the acetic acid concentration, [acid], decreases
proportionately because each increment of base converts 0.01 mole of
acetic acid into 0.01 mole of sodium acetate according to the reaction
HAC + NAOH NaAc + H,0 2
(0.1-0.01) (0.01) (0.1+ 0.01)

2. AMore Exact Equation for Buffer Capacity

The buffer capacity calculated from equation (1) of buffer capacity.
" It gives the average buffer capacity over the increment of base added.
Koppel and Spiro and Van Slyke developed a more exact equation,

K,[H,0+]
Bmax = 2.3C ,(K¡+[H,0*)² 3

Where Cisthe total buffer concentration, that is, the sum of the molar
concentrations of the acid and the salt.

3. The Influence of Concentration on Buffer Capacity

The buffer capacity is affected not only by the [Salt]/[Acid] ratio but also by
the total concentrations of acid andsalt.
4. Maximum Buffer Capacity
An equation expressing the maximum buffer capacity can be derived from
the buffer capacity formula of Koppel and Spiro and Van Slyke equation
(3).
The maximum buffer capacity occurs where pH = pKa
 In equivalent terms, where [H,0]=k,:
Substituting [H,0 ]for k, in both the numerator and the denominator of
equation (3) gives.
where C is the total buffer concentration.

Baax 2.303C (H,0*

(2H,0»)'
2.303
C Bmax = 0.576C
Pmast 4
5. Neutralization Curves and Buffer Canacity
Buffer capacity can be obtained by considering the titration curves of
strong and weak acids when they are mixed with increasing quantities
of alkali.

The reaction of an equivalent of an acid with an equivalent of a base is

called neutralization.
" It can be expressed according to the method of Bronsted and Lowry.

D
BUFFERS IN P'CEUTICAL & BIOLOGICAL SYSTEMS
& Pharnaceutical
Buffer solutions are used frequently in pharmaceutical practice,
particularly in the formulation of ophthalmic solutions.
They also find application in the colorimetric determination of pH.

Gifford15 suggested two stock

solutions - one containing boric acid
and the other monohydrated sodium
carbonate, which, when mixed in
various proportions, yield buffer
solutions with pH values from about5
 A
buffer system introduced by Palitzsch and modified by Hind and Goyan
consistsof boricacid, sodium borate, and sufficient sodium chloride to
make the mixtures isotonic
Itis used for ophthalmic solutions in the pH range of 7 to 9.
The buffers of Clark and Lubs based on the original pH scale of
Sorensen, have been redetermined at 25°C by Bower and Batesl8 so as
to conform to the present definition of pH.
The Clark-Lubs mixtures and their corresponding pH ranges are as
follows:
a) HCl and KCl,pH 1.2 to 2.2
b) HCl and potassium hydrogen phthalate, pH 2.2 to 4.0
c) NaOH and potassium hydrogen phthalate, pH 4.2 to 5.8
d) NaOH and KH,PO4, pH 5.8to 8.0
e) H,B03, NaOH, and KCI, pH 8.0 to 10.0
º General Procedures for Preparing Pharmaceutical Buffer
Solutions
a) Select a weak acid havinga pK, approximately equal to the pH at which the
buffer is to be used. This will ensure maximum buffer capacity.
b) From the buffer equation, calculate the ratio of salt and weak acid required
toobtain the desired pH. The buffer equation is satisfactory for approximate
calculations within the pH range of 4 to 10.
c) Consider the individual concentrations of the buffer salt and acid needed to
obtain a suitable buffer capacity. A concentration of 0.05 to 0.5 M is
usually sufficient, and a buffer capacity of 0.01 to 0.1 is generally
adequate.
d) Choice of a pharmaceutical buffer include availability of chemicals,
sterility of the final solution, stability of the drug and buffer on
aging,cost of materials,and freedom from toxicity.
 Sorensen proposed a mixture of the salts of sodium phosphate for
buffer solutions of pH 6to 8.
Sodium chloride is added toeach buffer mixture to make it isotonic
with body fluids
For example, a borate buffer, because of its toxic effects,certainly cannot
be used to stabilize a solution to be administered orally or parenterally.
e) Finally, determine the pH and buffer capacity of the completed buffered
solution usinga reliable pH meter.
*In vivo hiologic Buffersystem (hiological system)
Blood
Blood consists of primary (plasma) and secondary buffer
Erythrocytes) systems contributing the pH 7.4.
When the pH of the bloods below 7.0 or above 7.8, life is in danger. The
pHof the blood in diabetic coma is reported to drop as low as 6.8.
Primary buffers present in plasmna are carbonic acid-bicarbonate
system and acid/alkali salts of phosphoricacid system.
Secondary buffers that are present in erythrocytes are

Haemoglobin/oxyhemoglobin system and acid/alkali salts of

phosphoricacid system.
The buffer capacity is 0.0318 + 0.0035 for the whole blood, in which
0.031 is contributed by the cells and 0.008 is contributed by the plasma.
º Lacrimal fluids Lacrimalsac
Lacrimal gland
Lacrimal fluids (or tears) have
40
been found to have a great ExCretory ducts
of lacrimal gland

degree of affer
buffer capacity. Lacrimalcanal
Lacrmal punctum

allowing dilution of 1:15 with

Nasolacrimalduct
neutral distilled water.
 The pH of tears is about 7.4 with a range of 7.0 to 8.0.
Normally, pure conjunctivalfluid is moreacidic than the tear fluids
commonly employed in pharmacy.
The pH increases rapidly when the sample is removed for analysis
because of loss of carbon dioxide from the tear fluid.

º Urine:
The pH of urine is 6.0 for normal subjects (adults), when 24 h urine
was collected.

The pH may be as low as 4.5 oras high as 7.8.

If urine pH is low (4.5), hydronium ions are excreted into it urine by
the kidneys.
If urine pH is high (7.4), hydronium ions are retained by the action of
kidneys.
D
BUFFERED ISOTONIC SOLUTION
Isotonic buffered solution is defined as asolution which maintains
the isotonicity and the pH as that of the body fluids.
Isotonic buffer solution should be compatible with the body fluids for
the following reasons.
Blood and lacrimal fluids are in vivo buffer systems. Any
solution that comes in contact with these fluids should be buffered
to adesired pH, so that these are compatible with the body fluids.
Some solutions are meant for the application on delicate
membranes of the body. Such solutions may cause haemolysis,
tissue irritation, necrosis and tissue toxicity.
In such cases, solutions must be just to the same osmotic pressure
and tonicity as that of the body fluids.
 Applications:
Isotonicity should be adjusted for several dosage forms.
1. Parenteralpreparations shouldbe isotonic with blood plasma.
> There can be some flexibility depending on the route of administration
andQuantity of solution to be injected.
Intravenous infusions, irrigating solutions, lotions for open wound
Subcutaneous injection.
Parenteral preparations meant for diagnostic purposes, in order to
avoid false reaction

Solutions meant for intrathecal injection,because the volume of CSF

(Cerebro Spinal Fluid) is only 60 to 80 ml. Hence or hypotonic solutions
though in smallvolumes, will disturb the osmotic pressure and may
cause vomiting
2. Aqueoussolutions used as nasal drops.
3. Ophthalmicdrops
Preparation of Isotonic Buffer Solution
1) The drug and other ingredients are dissolved in water.
2) The pH of the solution is determined and adjusted to the desired
value.

3) The tonicity value of the solution is calculated procedures using

standard procedures.
4) The amount of sodium chloride required to adjust the tonicity is
calculated.
5) The required amount of sodium chloride is added to the solution, so
that the final solution becomes isotonic.
6) Isotonic diluting solution is added for maintaining the drug
concentration to the desired level (dose).
7) If the pH is also needs to be maintained, then buffered isotonic diluting
solution is added tomake up the desired volume (dose).
RINGERs
Ex- Isotonic diluting solutions are: SOLUTION

V
Isotonic sodium chloride solution
Dextrose solution
V Ringer solution

The isotonic buffered diluting solutions are available in acidic, neutra

and alkalinerange.
Isotonic buffered diluting solutions are
i) Iso-osmotic Solutions
Iso-osmotic solutions are those osmotic pressure as that of the cell
contents in question, but the solute is solutions which produce the same
permeable through the cell membrane thereby altering the tone of the
cell.

V Example of iso-osmotic solution is 1.8 % solution of urea.

ii) Isotonic Solution
Isotonicsolutions are those solutions which produce the same osmotic
pressure as that of the cell contents in question, with out net gain or
loss of both solutions,provided the cell membrane is impermeable to
the solutes.

Isotonic solutions are iso-osmotic as well as isotonic with the cells

and membranes.
Some of the standard isotonic solutions are:

0.9% w/v Normal saline (sodium chloride) solution

5.0% w/vDextrose solution
2.0% w/v Boricacid solution
 IsotonicSolution

ii) Hypertonic Solutions

Hypertonicsolutions are defined as those solutions containing the solute in
higher concentration than that is required for isotonic solutions.
Some hypertonic solutions are:
2.0% W/v Normal saline (sodium chloride) solution (concentration
>0.9% w/v).
10.0 % w/vDextrose solution (concentration > 5.0% w/v).
3.0 % w/v Boric acid solution (concentration > 2.0% w/v).

Hypertonic Solution

When red blood cells are suspended in a 2.0 % w/v solution of sodium
chloride, the water within the cells passes out through the cell
membranes in an attempt to dilute the surrounding salt solution.
Thisprocess continues until the salt concentrations on both sides of the
erythrocyte membrane are equal.
 Thus outward passage of water causes the cells toshrink and becomes
wrinkled or crenated. Such a salt solution issaid to be hypertonic with
respect to blood.

iv) Hypotonic Solution

Hypotonic solutions are defined as those solutions containing the
Solute in lower concentration than that is required for isotonic
solutions.

Hypotonic solutions are :0.2% w/v normal saline (sodium chloride)

When blood cells are suspended in a 0.2 % w/v solution of sodium
chloride (or in distilled water), water enters the blood cells causing
them to swell and finally burst with the liberation of haemoglobin.
This process is known as haemolysis.
Such a weak salt solution is said to be hypotonic with respect to blood.

Hypotonic Solution

Hypertonic Isotonic Hypotonic

3 Measurementof Tonicity
Isotonicity value is defined as the concentration of an aqueous
sodium chloride solution having same colligative properties as the
solution in question.
Apart from sodium chloride, anumber of chemicals and drugs
also included in the formulations. These ingredients also contribute to
the tonicity of the solution.
Two methods are mentioned below:
Hemolytic method: Red blood cells (RBCs) are suspended in various
drug solutions and the swelling of RBCs is observed bursting,
shrinking and wrinkling of the blood cells.
In hypotonic solutions, oxyhemoglobin is released, which is in direct
proportion to the number of cells hemolyzed.
In hypertonic solutions, the cells shrink and become wrinkled or
crenated

In isotonic solutions,the cells do not change their morphology.

Calculating Tonicity IIsing LËso Values
Freezing point depressions for solutions of electrolytes of both the weak
and strong types are always greater than those calculated from the
equation
ATF kc &new factor, L = K,introduced to overcome this difficulty

AT- Lc
The L value can be obtained from the freezing point lowering of
solutions of representative compounds of a given ionic type at a
concentration cthat is isotonicwith body fluids.
Thisspecific value of Lis written as Lise:
 The Ligo Value for a 0.90% (0.154 M) solution of sodium chloride, which
has a freezing point depression of 0.52°C andis thus isotonicwith body
fluids, is 3,4.
AT;
C
0.520
0.154=3.4

o Methods ofadjusting the tonicity &pH

CLASS-I
>These methods are of two types:
i. Cryoscopic method
ii. Sodium chloride equivalent method
CLASS-I
> These methods are of two types:
i. White Vincent method
i. Sprowls method

CLASS-I
i. Cryosconic method
>Principle:
Water has the freezing point of 0°C.
Blood contains a number of substances such as carbonic acid,
carbonates, salts of phosphoric acid and hemoglobin.
As a result, the depression in the freezing
point of the blood is -0.52 °C. When substances
such as sodium chloride are added to water

the freezing point of water decreases.

 ent of depression in the freezing point depends On t
oncentration of the added substance.
For example, sodium chloride at 1 % w v solution decreases th
freezingpoint of water to -0.58°C.
order make the drug solution isotonic, the freezing point

depression of the solution must be maintained at-0.52°C.

Initially the drug solution is prepared whosedepression in the freezing
point (AT) is known.
The remaining (AT;) value is adjusted by adding additional substances such
as sodium chloride. The freezing point depression of a number of drugs
are determined experimentally or theoretically a concentration of 10 %
w/v (or sometimes 0.5 % w/v).
Similarly the freezing point depression values of 1w/v solution of
sodiumchloride andother general ingredients are also determined.
Derivation:
Freezing point depression (AT) of blood 0.52°C.
" Since the drug solution must be isotonic, it must have AT;,same as that of
the blood.

AT,=0.52°C.
Totaldrug solution AT,= AT, of drug + AT, adjustingsubstance ..-.(1)
Freezingpoint depression (AT) of thetotal drug solution =0.52°C
AT, value of thedrug =xX AT, of 1 % drug solution =d
where x = drug concentration in the preparation, g/100 MI
AT,for adjusting solution =w Xa
where w=weight of the adjusting substance, g/100 mL.
a=AT; ofthe adjusting substance (sodium chloride), 19% (=0.58)
isotonic solution, equation (1) issubstituted.
 0.52°=d+ W,
The % w/v of adjusting substance needed is:
W= (0.52-d)/a = (0.52-d)/0.58 --------- (2)
Equation (2) is valid, if 1 % drug solution is specified. For any given
percentage strength of medicament (PSM), equation (2) may be
modified as:

W= [0.52- (PSM x d)]/ 0.58 -- ----3)

Thus, the desired concentration of adjusting substance is calculated and
added in order to make the drug preparation isotonic with blood.
Hence, if two or moresubstances are present, a sum of their freezing
point depression should be considered.
º Advantage:
Determination of depression in the freezing point is much simpler and
more convenient.

ii. Sodium Chloride Equivalent Method

Tonicic equivalent or sodium chloride equivalent method is used to
adjust the tonicity of pharmaceutical solutions.
Sodium chloride equivalent (D) of a drug is the amount of sodium
chloride that is equivalent to 1 gof the drug.
Equivalent refers to sodium chloride concentration having the same
osmotic effect as that of the drug.
In the absence of available data, the E value of a new drug can be
calculated from equation (4).
E= (17 X Ljo]/M -(4)
where M = molecular mass, AMU
Liso = freezing point depression of the drug solution for showing
isotonicity.
 Method:
The percent of sodium chloride required for adjusting isotonicity can
be calculated using equation (5).
PSA =0.9 - (PSM Eof medicament) ---oonoo-(5)
PSM= Percent strength of medicament
PSA = Percent of sodium chloride for adjustment of isotonicity
Equation (5) is used to calculate the amount of adjusting substance
(sodium chloride) required for making the solution isotonic. It is valid
for 100 mL solution.

CLASS-II
> These methods are oftwo types:
i. White Vincent method
ii. Sprowls method
i. White Vincent method
The class II methods of computing tonicity involve the addition of
to the drugs tomake an isotonic solution, followed by the addition
of an isotonic or isotonic-buffered diluting vehicle to bring the solution
to the final volume.

Stimulated by the need to adjust the pH in addition to the tonicity of

ophthalmic solutions, White and Vincent developed a simplified
method.

Suppose that one wishes to make 30 mL of a19% solution of procaine

hydrochloride isotonic with body fluid.

" First, the weight of the drug =W

" Multiplied bythe sodium chloride equivalent=E
0.3g x 0.21-0.063g (1)
Qty of sodium chloride osmotically equivalent to 0.3 g of procaine HCL
 Second, it is known that 0.9 g of sodium chloride, when dissolved
enough water to make 100 ml,yields asolution that is isotonic.
The volume V, of isotonic solution that can be prepared from 0.063 g
sodium chloride (equivalent to 0.3 g of procaine hydrochloride)
obtained by solving the proportion.
0.9g 0.063
100 ml (2)
100
V= 0.063 X (3)
0.9

V=7.0ML (4)
In equation (3)
Thequantity 0.063 is equal to the weight of drug w
Multiplied by the sodium chloride equivalent E, as seen in equation (1).
The value of theratio 100/0.9 is 111.1.
Accordingly,equation (3) can be written as
Here V=w xEx111.1 (5)
V= is the volume in milliliters of isotonic solution that may be
prepared by mixing the drug with water
weightin grams of the drug given in the problem
E=is the sodium chloride equivalent obtained from reference range.
The constant, 111.1, represents the volume in milliliters of isotonic
solution obtained by dissolving 1g of sodium chloride in water.
V=0.3*0.21x111.1 (5)
v=7.0ml
ii. Sprowls method
V=0.3gxEx111.1
NOTE:- The-value in Sprowl method is based on 1fluid ounce of the preparation
After calculating the volume of water to render the drug isotonic dilution
solution follow to make the volume of the preparation
The usedto compute the amount of isotonic diluting solution or vehicle
V.o=Vof Rx -V of water