Isotonic and Buffer solutions

Objectives
1. Apply the buffer equation, also known as the
Henderson-Hasselbach equation, for a weak acid or
base and its salt.
2. Discuss the factors influencing the pH of buffer
solutions.
3. Understand the concept and be able to calculate buffer
capacity.
4. Discuss the relationship between buffer capacity and
pH on tissue irritation.
5. Discuss the concept of tonicity and its importance in
pharmaceutical systems.
6. Calculate solution tonicity and tonicity adjustments. 2
 Introduction
• Buffer solutions
• solutions that resist change in Hydronium
ion and the hydroxide ion concentration
(and consequently pH) upon addition of
small amounts of acid or base, or upon
dilution.
• Consist of a weak acid and its conjugate
base (more common) or a weak base and
its conjugate acid (less common).
• 3
 Definition
• Nonelectrolytes, the solution will contain only molecules, and
the osmotic pressure will vary only with concentration of the
solute.
• Electrolytes: solution will contain ions, and the osmotic
pressure of the solution will vary not only with the
concentration but also with the degree of dissociation of the
solute.
• Isosmotic solutions: solutions that have the same osmotic
pressure
• Isotonic solution: a solution having the same osmotic pressure
as a specific body fluid
• Hypotonic solution: a solution of lower osmotic pressure
than that of body fluids.
• Hypertonic solution: a solution of higher osmotic pressure
than that of body fluids.
4
• Important for the pharmacist for preparation of
ophthalmic, nasal, parenteral and some rectal
preparations
• Drugs as buffers:
• Salicylic acid soln in a soft glass bottle is
influence by the alkalinity of the glass.
• Henderson-Hasselback equation

5
 Tonicity
– extent of swelling or contraction of biological
membrane (cells, mucous membranes)

– cell membranes are semipermeable

– hypertonic = higher than cells

• causes cells to crenate or shrink

– hypotonic = lower than cells

• causes cells to rupture (lyse)

– isotonic = same (isoosmotic)

7
 Measurement of Tonicity
• Two methods
• 1. Hemolytic method
• 2. Based on Methods used to determine
colligative properties.
 Hemolytic method
• Suspend the RBC in solutions.
• Observe the effect of various solution of
drug on appearance of RBC.
• Hypotonic solutions liberate
oxyhemoglobin in direct proportion to the
number of cells hemolysed.
• By such means the van’t Hoff i (π=iRTC)
can be determined and the value
compared with that computed from
cryoscopic data, osmotic co-efficient, and
activity co-efficient.
 Based on Methods used to
Determine Colligative properties
• This method is based on the measurement of slight
temp differences in the vapor pressure of thermally
insulated samples contained in constant humidity
chambers.
• The freezing point of blood is -0.56 oC and of tear is -
0.80 oC.
• Now for both it is -0.52 oC.
• This temp corresponds to the freezing point of 0.9%
Nacl solutions, which is therefore considered as a
isotonic with both blood and lacrimal fluids
Calculations for preparation of isotonic
solution:
• Freezing point depression (colligative properties)
- 0.52 is the freezing point of both blood serum and lacrimal fluids

• For nonelectrolytes (negligible dissociation) as 1gm of molecular weight of

any nonelectrolyte. The freezing point of the solution is about 1.86 C below
the freezing point of water
• Therefore: we can calculate the weight of any non electrolyte that should be
dissolved in each 1000g g of water if the solution is to be isotonic with body
fluids
• Boric acid: MWt 61.8 thus if 61.8 g in 1000 g of water should produce a
freezing point of -1.86 oC

1.86 C  61.8 g 

•  X = 17.3 g
0.52 C  X g 
• So 17.3 g of boric acid in 1000 g of water (1.73 %) should make a solution
isotonic with lacrimal fluid. 11
• For electrolytes,
• Osmotic pressure depends more on the number than on the
kind of particles, substances that dissociate have a tonic effect
that increases the dissociation:
• The greater the dissociation, the smaller the quantity
required to produce any given osmotic pressure/
• it depends on the degree of dissociation
• Example;
• NaCl (M.Wt. 58.5) in weak solutions is 80 % dissociated, then
each 100 molecules yields 180 particles, 1.8 times
nonelectrolyte. This dissociation factor is symbolized by the
letter i .
1.86 C 1.8 58.5 g 

0.52 C  X g 
• X = 9.09 g
• So 9.09 g of NaCl in 1000 g of water (0.9 % w/v) should make
a solution isotonic with blood or lacrimal fluid.
12
• Isotonic solutions are calculated by the following formula

0.52  molecular weight

 g of solute per 1000 g of water
1.86  dissociati on i 
• The value i for many medicinal salts has not been experimentally determined
• Some salts as zinc sulfate with 40% dissociation and i value = 1.4 are
exceptional.
• Most medicinal salts approximate the dissociation of NaCl.
• If the number of ions is known so:
• Nonelectrolytes and substances of slight dissociation i =1
• Substances that dissociate into 2 ions i = 1.8
• Substances that dissociate into 3 ions i = 2.6
• Substances that dissociate into 4 ions i = 3.4
• Substances that dissociate into 5 ions i = 4.2
13
• The Sodium Chloride Equivalent (E value) of a
drug: is the amount of sodium chloride which has the
same osmotic effect as 1 gram of the drug.
Mwt of NaCl i factor of subs tan ce
  Sodium Chloride equivalent
i factor of NaCl Mwt ofsubs tan ce

• How much NaCl should be used in preparing 100 ml

of 1% w/v solution of atropine sulfate, which is to be
made isotonic with lacrimal fluids?
695  2.6 1g 

58.5  1.8 X g 
• M.Wt of NaCl = 58.5, i = 1.8
• M.Wt of atropine sulfate = 695, i = 2.6
14
 Methods of adjusting tonicity
and pH
• Class I
A. CRYOSCOPIC METHOD/ FREEZING PT
DEPRESSION METHOD
B. SODIUM CHLORIDE EQUIVALENT
METHOD
Class II
A. WHITE- VINCENT METHOD
B. SPROWLS METHOD
• 15
 Methods for adjusting Tonicity &
• Two type pH
• 1. Class I methods : NaCl or another substance is
added to the solution of the drug to lower the
freezing point of solution to -0.52 oC and thus
make isotonic with body fluid.
• E.g. Cryoscopic Method(used of freezing point)
• NaCl equivalent method (E-value)
• 2. Class II methods: water is added to the drug in
sufficient amount to form isotonic s0lution. The
preparation is then brought to its final vol with
isotonic or buffered isotonic dilution solution.
• E.g. White Vincent method and Sprows
 Cryoscopic Method

• Freezing point is used isotonicity

calculations when the agent has tonicic
effect and does not penetrate the biologic
membranes (eg.RBC)
• -0.52C- Fp of both blood and lacrimal
fluids thus also isotonic solution.
• Each pharmaceutical substance has own
data in Fp depression at strength of 1%
 FREEZING POINT DATA FOR SELECTED AGENTS FREEZING
POINT DEPRESSION,AGENT 1% SOLUTIONS (T1%ƒ )

• Atropine sulfate 0.07

• Boric acid 0.29
• Butacaine sulfate 0.12
• Chloramphenicol 0.06
• Chlorobutanol 0.14
• Dextrose 0.09
• Dibucaine hydrochloride 0.08
• Ephedrine sulfate 0.13
• Epinephrine bitartrate 0.10
• Ethylmorphine hydrochloride 0.09
• Glycerin 0.20
• Homatropine hydrobromide 0.11
• Lidocaine hydrochloride 0.063
• Lincomycin 0.09
• Morphine sulfate 0.08
• Naphazoline hydrochloride 0.16
• Physostigmine salicylate 0.09
• Pilocarpine nitrate 0.14
• Sodium bisulfite 0.36
• Sodium chloride 0.58
• Sulfacetamide sodium 0.14
• Zinc sulfate 0.09
18
 Problem
• How many milligrams each of NaCl and
dibucaine HCl are required to prepare 30
ml of a 1% solution of dibucaine HCl (1%-
0.08-Fp lowering)isotonic with tears?
• Note: To make the soln isotonic, the Fp
must be lowered to -0.52C
A. (from the table) It is determined that a 1%
solution of dibucaine HCl has a freezing
point lowering of 0.08%.
B. Thus, sufficient NaCl must be added to
lower the freezing point 0.44 (0.52-0.08) 19
 it is determined that a 1% solution of
sodium chloride lowers the freezing point by
0.58. By proportion:
• 1% (NaCl) = 0.580
• x% (NaCl) 0.440
• x = 0.76% (the concentration of sodium
chloride needed to lower the freezing point
by 0.44,required to make the solution
isotonic)

20
C. Thus to make the 30ml soln

30ml X 1%= 0.3 g=300mg dibucaine HCl

30ml X 0.76%=0.228g=228mg sodium

chloride

21
 NaCl equivalent
• For determining the amount of sodium chloride used to cause a solution
isotonic,
• multiply the quantity of each drug in the prescription by it’s sodium chloride
equivalent E ,
• and subtract this value from the concentration of sodium chloride which is
isotonic with body fluids (0.9 gm per 100 ml).
• How many grams of sodium chloride should be used in compounding the
following prescription
• R/ Pilocarpine nitrate 0.3 g
• Sodium chloride q.s.
• Purified water 30 ml
• Make isoton. Sol.
• Sig. for the eye
Dr. Osama A. A. Ahmed 22
 Isotonic and Buffer solutions
• Calculations for preparation of isotonic solution:
Example: How many grams of sodium chloride should be used in compounding
the following prescription
R/ Pilocarpine nitrate 0.3 g
Sodium chloride q.s.
Purified water 30 ml
Make isoton. Sol.
Sig. for the eye
• Sod. Chloride equivalent for Pilocarpine nitrate = 0.23
• 1- 0.23 X 0.3 = 0.069 g of NaCl represented by the pilocarpine nitrate
• 2- 0.9 g NaCl 100 ml water to be isotonic
Xg 30 ml
X = 0.9 X 30 / 100 = 0.27 g
• 3- 0.27 - 0.069 = 0.201 g of sodium chloride to be used
Dr. Osama A. A. Ahmed 23
EXAMPLES:
Rx
1.A solution contains 3.0 g of Ephedrine sulfate in 75
ml.
1.How many NaCl must be added to make the
solution isotonic?
2.How much Dextrose would be required to make
the solution isotonic?
E- value
Ephedrine sulfate 0.13
Dextrose 0.16

24
a.
Step 1: 3 g x 0.13 = 0.39 g of NaCl represented by
Ephedrine sulfate
Step 2: 0.009 x 75 ml = 0.675 g of NaCl in 75 ml of an
isotonic solution
Step 3: 0.675 g – 0.39 g = 0.285 g of NaCl to be used or
added to make the solution isotonic.
b.
Step 4:
= g of NaCl to be added (step 3 answer)
E value of dextrose
= 0. 285
0.16
= 1.78 g of Dextrose is required to make the
solution isotonic 25
2. How many grams of NaCl should be used in
compounding the following prescription?
Rx E value
Phenacaine HCl 1.5% 0.20
Chlorbutanol ½% 0.24
NaCl qs
P. water ad 60 ml

Make the soln isotonic

Sig one drop in each eye

26
Step 1: (.015 x 75 ml) x 0.20 = 0.225 g of NaCl represented by
Phenacaine HCl
( 0.005 x 75 ml) x 0.24 = 0.09 g of NaCl represented by
Chlorbutanol
0.315 g of NaCl rep by
Phenacaine & Chlorbutanol
Step 2: 0.009 x 75 ml = 0.675 g of NaCl in 75 ml of an isotonic
solution
Step 3: 0.675 g – 0.315 g = 0.36 g of NaCl to be used or added to
make the solution isotonic.
Step 4:
= g of NaCl to be added (step 3 answer)
E value of boric acid

= 0.36 g/ 0.50

= 0.72 g of boric acid

27
 White –Vincet Method
• water is added to the drug in sufficient amount to
form isotonic s0lution.
• The preparation is then brought to its final vol
with isotonic or buffered isotonic dilution
solution.
• So the eqn V = W * E * 111.1
• Where V is vol in ml of isotonic solution that may
be prepared by mixing the drug with water.
Rx

E - value
Phenacaine HCl 0.06 0.20
Boric acid 0.3 0.50
H20 q.s 100 ml
V = wt x E x 111.1
V = [ (0.06 g x 0.20) + (0.3 g x 0.50) ] x 111.1
V = 18 ml
The drugs are mixed with water to make 18 ml of an
isotonic solution, and the preparation is brought to a
volume of 100 ml by adding an isotonic diluting solution.

29
 The Sprowls Method
• V value
• Volume of the water to be added to a specified
weight of the drug (0.3 or 1.0g depending on the
table used)-30 ml or 1 Fz.
• The basic principle underlying the use of V values
to prepare an isotonic soln. of the prescribed drugs
and then dilute this solution to final volume with
suitable isotonic vehicle
 • The eqn V = 0.3 * E * 111.1 could be used to
construct a table of values of V when the wt of
the drug is fixed.
• Sprowls choose the wt of drug 0.3 g, the
quantity for 1 fluid ounce of 1% solution.
• Compute the vol V of isotonic solutions of 0.3
g drug with sufficient water for drugs
commonly used in ophthalmic and parental

Dr. Osama A. A. Ahmed 31

 Problems
1.
• Dexamethasone Sodium Phosphate– 1%
• Purified water qs ------ 30ml
• MFt isotonic solution

Dr. Osama A. A. Ahmed 32

 Buffered isotonic solutions
 Pharmaceutical solutions that are meant for
application to delicate membrane of the body
should be adjusted to same osmotic pressure as
that of body fluids.
 Isotonic solutions cause no swelling or
contraction. E.g. isotonic NaCl solutions.
 Mix small quantity of blood with aq. NaCl
solutions of varying tonicity.
 Blood cells + 0.9 % NaCl = cells retain normal
size (Isotonic with blood)
 Blood cells + 2 % NaCl = cells shrink and become
wrinkled or crenated (Hypertonic with blood)
 Blood cells + 0.2 % NaCl = cells swells and burst
liberating hemoglobin (Hypotonic with blood)
 Buffered isotonic solutions
 The RBC membrane permit the passage of water
molecules, urea, ammonium chloride, alcohol, boric
acid.
 A 2.0 % boric acid solution is isosmotic to blood cell.
 The molecules of boric acid pass freely through the
erythrocyte membrane regardless of conc.
 As a result boric acid solution is hypotonic and cause
hemolysis.
 So the solution containing quantity of drug calculated to
be isosmotic with blood is isotonic only when blood cells
are impermeable to solute molecules and permeable to
solvent molecules.
 Mucous lining of the eye is true semi permeable to boric
acid solutions and hence 2 % boric acid solution is
isotonic ophthalmic preparation.
 Factors affecting the pHof the
buffer equation
1. Addition of neutral salts to buffer changes
the pH of the solution by altering ionic
strength
2. Dilution
3. Temperature
Ex.
Acetate buffer-pH increased with temp
Basic acid sodium borate buffer-decrease
w/temp 35
 Buffers in Pharmaceutics and
biological system
A. In Vivo Biological buffer system

Blood is maintained at a pH of about 7.4

1o buffer-plasma
2o buffer-erythrocyte

36
 Plasma
• Contains carbonic acid/carbonates and
acid/alkali sodium salts of the phosporic
acid as buffers

• Behave as acids in blood can combine

with base and can act as buffer

37
 Erythrocytes
• The two buffer consist of
hemoglobin/oxyhemoglobin and acid/alkali
of potassium salts of phosphoric acid.

• Great degree of buffer capacity

• Dilution of 1:15 with neutral distilled water
before alteration of pH is noticed
• pH=7.4 (7.8) or slightly higher 38
 Pure conjunctival fluid
• More acidic than tear fluid
• Usually used in pH measurements

• Urine :
The urine of a normal adult has a pH of
about 6.0 with the range of 4.5 to 7.8
• When the pH of the urine is below normal
values, hydrogen ions are excreted by the
kidneys.
 Pharmaceutical buffers
• Ophthalmic solns
• Calorimetric determination of pH
• Research studies in w/c pH must be held
constant

40
A. Gifford (boric acid and other
monohydrated sodium carbonate)pH(5-9)
when mixed
B. Sorensen (mixture of salts of sodium
phosphate-pH 6-8) NaCl is added to
each buffer mixture to make it isotonic
with body fluids
C. Palitzch modified by Hind and Goyan
Consist of boric acid, sodium borate and qs
NaCl to make it it isotonic
Used for ophthalmic soln (pH range of 7-9)
41
Influence of buffer capacity and
pH on tissue irritation
• If the pH is greatly removed from the
normal pH of the relevant body fluid

• Formulating ophthalmic soln

• Parenteral products
• Fluids to be applied to abraded surfaces

42
 Tissue irritation
• Due to large pH differencesbetween the
soln being administered and the
physiological environment in w/c it is used
will be minimal
• According to: Friedwald, Huger and
Heuman
• pH in the eye should be 4.5-11.5
• w/out mark pain and damage
43
 Buffered isotonic soln
• Cause no swelling
• No contraction of the tissues
• No discomfort when instilled into the eye,
nasal tract, blood stream or other body
tissues
• Ex. Isotonic sodium chloride

44
 BUFFER EQUATION
• Buffer Equation for a weak acid and its
salt

• pH = pKa + log [salt]

[acid]

45
 Problems:
1.What is the molar ratio (salt/acid), required
to prepare an acetate buffer of pH 5.0 Also
express the result in mole percent. (pKa=
4.76)
A. Mole ratio?
B. Mole percent?

46
Data: pH = 5.0 pKa= 4.76
A.
Eq: pH = pKa + log [salt]
[acid]

5 = 4.76 + log [ salt ]

[acid ]

5 – 4.76 = log [ salt ]

[ acid ]

antilog 0.24 = [ salt ]

[acid ]

1.74 = [ salt ]
[acid ]
The mole ratio of salt to acid is 1.74/1.
47
•
 B.
Mole ratio of salt to acid is 1.74/1
Salt: 1.74
Acid: + 1
_____
salt + acid 2.74
Mole fraction of salt in salt-acid mixture = 1. 74 = 0.635
1 + 1.74
Mole percent = mole fraction X 100
= 0.635 x 100
= 63.5 %

Dr. Osama A. A. Ahmed 48

Problem no.2

A buffer solution was prepared by mixing 0.25M

CHOONa and 0.125M HCOOH (pKa =3.75).
A. Determine the pH of the buffer solution
B. If the same composition is to be used in
preparing a buffer solution with a pH of 4.5.
What will be the molar ratio of the salt to
acid?
a.What is the percentage of the ionized
species in the buffer system
b.What is the percentage of the unionized
species in the buffer system
49
A.
pH = pKa + log [salt]
[acid]
pH = 3.75 + log 0.25 M
0.125 M
pH = 3.75 + log 2
pH = 3.75 + 0.30
pH = 4.05

50
B.
pH = pKa + log [salt]
[acid]
4. 5 = 3.75 + log [salt]
[acid]
4.5 – 3.75 = log [salt]
[acid]
antilog 0.75 = [salt]
[acid]
5. 6 = [salt]
[acid]
51
The ratio of salt to acid is 5.6:1.
B 1 : percentage of the ionized species in the buffer system
The ratio of salt to acid is 5.6 : 1.
Salt (ionized) : 5.6
Acid ( unionized) : 1
_____________
6.6
% of ionized species = ionized______ x 100
ionized + unionized
= 5.6 x 100
6.6
= 84.85 %

B2
% of unionized species = unionized_____ x 100
ionized + unionized
= 1_ x 100
6.6
= 15. 15 %

52
 3. A buffer was prepared containing 5 x 10 -2 M
Sodium borate with 5 x 10 -7 M Boric acid. Determine the
Ka of the acid if the solution have a pH of 1.0 .
pH = pKa + log [salt]
[acid]
1.0 = pKa + log 5 x 10-2
5 x 10 -7
1. 0 = pKa + 5
1-5 = pKa
-4 = pKa
since pKa = - log Ka
antilog - 4 = Ka
1 x 10 -4 = Ka
53
Buffer Equation for a weak base
and its salt

pH = pKw - pKb + log [base]

[salt]

54
Problems:
1.What is the pH of a solution containing 0.10
moles of ephedrine and 0.01 mole of ephedrine
hydrochloride per liter of solution? The pKb of
ephedrine is 4.64.
pH = pKw - pKb + log [base]
[salt]
pH = 14 – 4.64 + log 0.10
0.01
pH = 14 – 4.64 + 1

= 10.36 55
1.The Kb of pilocarpine is found to be 6.34 x l0 -7 at 25°C.
1.Determine the mole ratio (salt/base) present at pH of 9
2.Deterrnine the mole percent of the base
3.Determine the mole percent of the salt

a. mole ratio (salt/base) present at pH of 9

pKb = - log Kb
= - log (6.34 x l0 -7)
= 6.20

56
 pH = pKw - pKb + log [base]
[salt]
9 = 14 – 6.20 + log [base]
[salt]
9 = 7. 8 + log [base]
[salt]
9 – 7.8 = log [base]
[salt]
1.2 = log [base]
[salt]
antilog 1.2 = [base]
[salt]
15.85 = [base]
[salt]
The mole ratio of salt to base is 1: 15.85.
Jham 57
b. mole percent of the base
salt : 1
base : + 15.85
____
16.85
mole percent (base) = 15.85 x 100 = 94.07%
16.85
c. mole percent of the salt
mole percent (salt) = 1_ x 100 = 5.93%
16.85

58
 Buffer capacity
• The magnitude of the resistance of a
buffer to changes is referred to as a
buffer capacity β.
• Also known as a buffer efficiency, buffer
index, buffer value.
• It is the ratio of the increment of strong
acid or base to the small changes in pH
brought about by addition.
• β = Δ B/ ΔpH where Δ B is small
increment in gram equivalent / liter of
strong base added.
BUFFER CAPACITY
β β/ pH

Where:
β is the small increment in gram
equivalent/liter of strong base added
to the buffer solution
pH is the change in pH

60
• Calculate the approximate buffer capacity of the given data:
pH of glacial acetic acid and sodium acetate solution: 6.9
pH of buffer solution added with 0.05 N NaOH = 8.4

β = β
pH
= 0.05
8.4 – 6.9
= 0.033

61
• Maximum Buffer Capacity
• βmax = 0.576 C where C is the total
buffer concentration

• Example
• What is the maximum buffer capacity of an
acetate buffer with a total concentration of
0.020 mole/liter?
• βmax = 0.576 X 0.020
• = 0.01152 or 0.012
62