Buffered and

Isotonic Solutions

Contents
The Buffer Equation
Buffer Capacity
Buffers in
pharmaceutical and Biologic Systems
Buffered Isotonic Solutions
Methods of Adjusting Tonicity and pH
2

Introduction
Buffered Solutions ?

Buffered Solutions
0.1N HCl 1ml

H 2O

NaCl

HAc,NaAc

pH7

pH7

pH4.7

4.58
buffer action
4

Buffered Solutions
Combination of a weak acid and its conjugate base

HA + OH-

A- + H2O

Combination of a weak base and its conjugate acid

A-

H3O+

HA

+ OH5

Contents
The Buffer Equation
Buffer Capacity
Buffers in
pharmaceutical and Biologic Systems
Buffered Isotonic Solutions
Methods of Adjusting Tonicity and pH
6

The Buffer Equation

A Weak Acid and Its Salt
HAc + H2O

H3O+ + Ac-

K1[HAc][H2O] = K2[H3O+][Ac-]
Ka =

[H3O+][Ac-]
[HAc]

salt
acid

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

7

The Buffer Equation

A Weak Acid and Its Salt

pH= pKa+log

Dissociation
exponent

[salt]
[acid]

Buffer equation or
Henderson-Hasselbalch
equation
8

Common ion effect

* when Sod. acetate is added to acetic acid

[H3O+][Ac-]
Ka =
[HAc]
is momentarily disturbed since the acetate ion supplied
by the salt increases the [Ac-]

HAc + H2O

H3O+ + Ac-

The ionization of HAc is repressed upon the addition

of the common ion [Ac-]
9

The Buffer Equation

A Weak Base and Its Salt
OH- + BH+

B + H2O
Kb =

[OH-][BH+]
[B]

salt
base

10

The Buffer Equation

A weak base and its salt
[OH-] = Kb

[base]
[salt]
[H3O+] [OH-] = Kw

-log[H3O+]= - logKw log1/Kb - log[salt]/[base]

11

The Buffer Equation

A Weak Acid and Its Salt

pH= pKw- pKb + log

[base]
[salt]

* Buffers are not ordinarily prepared from weak

bases because of the volatility & instability of the

bases and because of the dependence of their pH
on pKw, which is often affected by temp. changes.
12

Activity coefficients
HAc + H2O
O+][Ac-]

[H3
Ka =
[HAc]

Conc.

H3O+ + Ac-

aH3O+ aAc=
aHAc

activity

activity
Molar conc.

(H3O+cH3O+)(Ac- CAc-)
=
(HAcCHAc)
Activity coefficients
13

Activity coefficients
- log[aH3O

+]

= - log Ka + log

aAcaHAc

* activity coefficient of the undissociated acid HAc is

essentially 1 and may be dropped.

pH = pKa + log

[salt]
[acid]

+ log Ac-

14

pH
1. Altering the ionic strength
Addition of neutral salts
Dilution (alter activity coefficients)
2. Temperature
The pH of the most basic buffer was found
to change more markedly with temp. than
that of acid buffers, owing to Kw.
15

pH indicator
Acid indicator

HIn + H2O

H3O+ + In-

Acid color

KIn =

Alkaline color

[H3O+ ][ In-]

[HIn]

base
acid

16

PH indicator
pH = pKIn + log

[base]
[acid]

1/10~10/1

* From experience, one cannot discern a change from the

acid color to the salt color the ratio of [base] to [acid] is
about 1 to 10
* The effective range of the indicator is

pH =pKIn + 1
base

10/1

1/10

acid
17

pH indicator
Characteristics of colorimetric method
less accurate
less convenient but less expensive than
electrometric method
difficult to apply for the unbuffered
pharmaceutical preparation (change the pH indicator itself is acids or base)
error may be introduced by the presence of
salts & proteins
18

Contents
The Buffer Equation
Buffer Capacity
Buffers in
pharmaceutical and Biologic Systems
Buffered Isotonic Solutions
Methods of Adjusting Tonicity and pH
19

Buffer capacity
the magnitude of the resistance of a buffer to
pH changes

=
buffer capacity
= buffer efficiency
= buffer index
= buffer value

B
pH

B : small increment in gram equivalents/Liter

of strong(or acid) added to the buffer
soln. to produce a pH change of pH
20

Buffer capacity ( )
HAc + NaOH
(0.1- 0.01)

0.01

NaAC

+ H2O

(0.1+ 0.01)

Before the addition of NaOH

[salt]
pH=pKa + log
[acid] = 4.76
After the addition of NaOH
pH=pKa + log [salt] + [base] = 4.85
[acid] - [base]

B
0.01
=
= 0.11
pH
0.09
21

Buffer capacity
A more exact equation for buffer capacity (1914, 1922)
= 2.3 C

Ka [H3O+]
(Ka + [H3O+])2

c : total buffer conc.(sum of the molar conc. of

the. acid & the salt)

---- at any [H3O+]

22

Maximum Buffer capacity

max occurs where pH = pKa ([H3O ] = Ka)
+

[H3O+]2

2.303
max = 2.303 C
C
=
+
2
4
(2 [H3O ])

max = 0.576 C
( pH = pKa )
23

Characteristics of Buffer Capacity

is not a fixed value, but rather depend on the amount
of base added
depends on the value of the ratio [salt]/[acid] and
magnitude of the individual concentrations of the
buffer components
The greatest capacity(max) occurs where
[salt]/[acid] = 1 and pH = pKa
Because of interionic effects, buffer capacities do not in
general exceed a value of 0.2
24

Universal Buffer
Total buffer capacity of a universal buffer
(combination of several buffers)

25

Contents
The Buffer Equation
Buffer Capacity
Buffers in
pharmaceutical and Biologic Systems
Buffered Isotonic Solutions
Methods of Adjusting Tonicity and pH
26

In Vivo biologic buffer systems

Blood
Primary buffers : Plasma ;
NaHCO3-- H2CO3, NaHPO4--NaH2PO4, protein
Secondary buffers : Erythrocytes ;
hemoglobin-oxyhemoglobin, K2Hpo4--KH2PO4
Lacriminal fluid
- pH: 7.4 (range 7 8 or slightly higher)
Urine
- pH: 6.0 (range 4.5 7.8)
- below normalhydrogen ions are excreted by the kidney.
- above pH 7.4hydrogen ions are retained by action of the kidney.
27

Pharmaceutical buffers
ophthalmic soln.
colormetric determination of pH
research studies in which pH must be
held constant
28

Pharmaceutical buffers
Clark-Lubs mixtures and pH
(a) HCl & KCl, pH 1.2 - 2.2
(b) HCl & potassium biphthalate, pH 2.2 - 4.0
(C) NaOH & potassium biphthalate, pH 4.2 - 5.8
(d) NaOH & KH2PO4 , pH 5.8 - 8.0
(e) H3BO3, NaOH & KCl, pH 8.0 - 10.0

29

Preparation of pharmaceutical buffer solutions

Steps for development of a new buffer
Select a weak acid having a pKa approximately equal
to the pH at which the buffer is to be used.
Calculate the ratio of salt & weak acid required to
obtain the desired pH.
Consider the individual conc. Of the buffer salt & acid
needed to obtain a suitable buffer capacity
* Individual conc. : 0.05 ~ 0.5M
* buffer capacity : 0.01 ~ 0.1
30

Preparation of pharmaceutical buffer solutions

Steps for development of a new buffer
Availability of chemicals, sterility of the final soln,
stability of the drug & buffer, cost of materials, freedom
from toxicity
ex) borate buffer toxic effect not be used for oral or
parenteral products.
Determine the pH and buffer capacity
using a reliable pH meter

31

Buffer in
pharmaceutical and biologic systems
Influence of buffer capacity and pH on tissue
irritation
* Tissue irritation will be minimal when

(a) Buffer solution , Volume

(b) Physiologic fluid - , Volume

32

Buffer in
pharmaceutical and biologic systems
Stability vs. optium therapeutic response
* Undissociated form of a weakly acidic or basic drug has a
higher therapeutic activity than the dissociated salt form.
* Molecular form is lipid soluble & can penetrate body
membranes readily, where the ionic form, not being lipid
soluble, can penetrate membranes only with greater
difficulty.
33

Buffer in
pharmaceutical and biologic systems
pH and solubility
* Influence of buffering on the solubility of base
- At a low pH : base is in the ionic form & usually very
soluble in aqueous media
- As the pH is raised : more undissociated base is formed
when the amount of base exceeds the limited water
solubility of this form, free base precipitates from soln.
Base soln. should be buffered at a sufficiently low pH
for stabilization against precipitation.
34

Buffer in
pharmaceutical and biologic systems
(Example)
GOAL: Compute the mole percent of
free base present on 25 and at a
pH of 7.4. The pKb of pilocarpine
is 7.15 at 25.

35

Buffer in
pharmaceutical and biologic systems
Example
C11H16N2O2 + H2O
(Pilocarpine base)

C11H16N2O2H + OH
(Pilocarpine ion)

[base]
pH= pKw- pKb + log [salt]
At pH 7.4

At pH 4.0

7.4 = 14 7.15 + log [base]

[salt]
[base] = 3.56 / 1
[salt]

[base]
4.0 = 14 7.15 + log [salt]
[base] = 0.0014 / 1
[salt]

Mole percent of base =

3.56 / (1 + 3.56) 100 =

Mole percent of base =

0.0014 / (1 + 0.0014) 100 =

78%

0.13%

36

Contents
The Buffer Equation
Buffer Capacity
Buffers in
pharmaceutical and Biologic Systems
Buffered Isotonic Solutions
Methods of Adjusting Tonicity and pH
37

Buffered isotonic solution

Red blood
cell

NaCl solution

2.0 %
0.9 %
0.2 %
Hypertonic, Isotonic Hypotonic,
Shrink
Hemolysis
38

Buffered isotonic solution

The term Isotonic should be restricted to
solutions having equal osmotic pressures
which respect to a particular membrane
(Husa)

Isotonicity valuethe concentration of an

aqueous NaCl soln. having the same
colligative properties as soln. (Goyan &
Reck)
39

Measurement of tonicity
Hemolytic method
apply red blood cells
based on the fact that a hypotonic soln. liberates oxyhemoglobin
in direct proportion to the number of cells hemolyzed

determine colligative properties (chapter 5)

modifications of the Hill-Blades Technique
based on a measurement of the slight temp. differences arising
from differences in the vapor pressure of thermally insulated
samples contained in constant-humidity chambers
Tf = 0.52 C (Freezing point lowering of
human blood & lacrimal fluid)
40

Calculating Tonicity Using Liso values

The Vant Hoff expression (Chapter 6)

Tf = L c
Liso =

Molal freezing
point depression
of water

Tf / c

0.52

Conc. that is
isotonic with
body fluids
41

Calculating Tonicity Using Liso values

42

Contents
The Buffer Equation
Buffer Capacity
Buffers in
pharmaceutical and Biologic Systems
Buffered Isotonic Solutions
Methods of Adjusting Tonicity and pH
43

Method of adjusting tonicity and pH

Class I add Sod. Chloride to lower the
freezing point of soln. to -0.52

Cryoscopic method
Sodium chloride equivalent method

Class II add Water to form an isotonic soln.

White-Vincent method
Sprowls method
44

Class I methods
Cryoscopic method ()
(Example)
How much NaCl is required to render 100mL of a 1% soln. of
apomorphine HCl isotonic with blood serum?
Tf0.9% of NaCl soln : 0.52(Isotonic with blood)
Tf1% of apomorphine HCl soln : 0.08 (from table)
to reduce the freezing point by an additional 0.44(0.52-0.08)
Tf1% of NaCl soln : 0.58
1(%)/X = 0.58/0.44 ; X = 0.76 (%)
Dissolve 1 g apomorphine HCl + 0.76g NaCl make 100mL
soln. with water
45

Class I methods
Sodium chloride equivalent(E) method
()

by Mellen & Seltzer

1g drug tonicity = Eg NaCl tonicity

E : weight of NaCl with the same freezing point depression
as 1g of the drug.

Tf = Liso c
Tf = Liso 1g/MW
3.4

c = 1 g / molecular
weight

58.45

E 17 Liso / MW

46

Class II methods
White-Vincent method
(Example)
GOAL: make 30mL of a 1% soln. of procaine
HCl isotonic with body fluid

47

Class II methods
Steps for White-Vincent method
Weight in grams of drug(0.3 g) Sod. Chloride
equivalent E(0.21..from table) = quantity of sod.
Chloride equivalent to w of drug(0.063 g)

0.9 g/100mL = 0.063 g / V

V = 0.063 100/0.9
V = 7.0 mL
Add isotonic-buffered diluting soln. to complete

V = w E 111.1
48

Class II methods
White vincent method

GOAL:
make 30mL of 1% soln.
of procaine HCl
isotonic with body
fluid

water

add 0.9%NaCl

0.9%NaCl
isotonic

0.3g drug
(E=0.21)

or

30ml

7ml
Isotonic buffered sol.

49

Class II methods
Sprowls method
w

E
V

0.9 g
=
100 ml

W = 0.3 g
(1% solution)

TABLE

?
50