Buffered and

Isotonic Solutions

1
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 ?

3
Buffered Solutions

0.1N HCl 1ml

H 2O NaCl HAc,NaAc

pH7 pH7 pH4.7

3 3 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 + OH-
5
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-]

[H3O+][Ac-] salt
Ka =
[HAc] acid

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

7
The Buffer Equation
A Weak Acid and Its Salt

[salt]
pH= pKa+log
[acid]

Buffer equation or
Dissociation Henderson-Hasselbalch
exponent 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
B + H2O OH- + BH+
salt
[OH-][BH+]
Kb =
[B] base

10
The Buffer Equation
A weak base and its salt
[base]
[OH-] = Kb
[salt]

[H3O+] [OH-] = Kw

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

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The Buffer Equation
A Weak Acid and Its Salt
[base]
pH= pKw- pKb + log
[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.
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Activity coefficients
HAc + H2O H3O+ + Ac-
activity
[H3 O+][Ac-] aH3O+ aAc-
Ka = =
[HAc] aHAc
Conc. activity
Molar conc.

(H3O+cH3O+)(Ac- CAc-)
=
(HAcCHAc)
Activity coefficients
13
Activity coefficients
- log[aH3O +] = - log Ka + log
aAc-
aHAc
* activity coefficient of the undissociated acid HAc is
essentially 1 and may be dropped.

[salt]
pH = pKa + log + log Ac-
[acid]

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.
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pH indicator
Acid indicator

HIn + H2O H3O+ + In-

Acid color Alkaline color

[H3O+ ][ In-] base

KIn = acid
[HIn]

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PH indicator
[base]
pH = pKIn + log 1/10~10/1
[acid]
* 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

B
= pH
buffer capacity
= buffer efficiency
= buffer index
= buffer value

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 NaAC + H2O
(0.1- 0.01) 0.01 (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
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Buffer capacity
A more exact equation for buffer capacity (1914, 1922)
Ka [H3O+]
= 2.3 C
(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 [H3O ]) 2 4

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

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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.

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Pharmaceutical buffers

ophthalmic soln.

colormetric determination of pH

research studies in which pH must be

held constant

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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

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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
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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

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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.

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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 C11H16N2O2H + OH
(Pilocarpine base) (Pilocarpine ion)
[base]
pH= pKw- pKb + log [salt]
At pH 7.4 At pH 4.0
[base]
7.4 = 14 7.15 + log [base] 4.0 = 14 7.15 + log [salt]
[salt]
[base] = 3.56 / 1 [base] = 0.0014 / 1
[salt] [salt]
Mole percent of base = Mole percent of base =
3.56 / (1 + 3.56) 100 = 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 Molal freezing
point depression
of water

Liso = Tf / c
Conc. that is
isotonic with
0.52 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 c = 1 g / molecular
weight
Tf = Liso 1g/MW
3.4 E 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
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Class II methods
White vincent method GOAL:
make 30mL of 1% soln.
of procaine HCl
water isotonic with body
fluid

0.9%NaCl add 0.9%NaCl

isotonic or 30ml
0.3g drug
(E=0.21) 7ml
Isotonic buffered sol.

49
Class II methods
Sprowls method
w E 0.9 g
=
V 100 ml

W = 0.3 g
(1% solution)
TABLE

?
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