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]

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The Buffer Equation
• A Weak Acid and Its Salt

[salt]
pH= pKa+log
[acid]

Buffer equation or
Dissociation Henderson-Hasselbalch
exponent equation
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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-]
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The Buffer Equation
• A Weak Base and Its Salt
B + H2O OH- + BH+
salt
[OH-][BH+]
Kb =
[B] base

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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
[H3O+][Ac-] aH3O+ •aAc-
Ka = =
[HAc] aHAc
Conc. activity
Molar
conc.
(γH3O+•cH3O+)•(γAc-• CAc-)
=
(γHAc•CHAc)
Activity
coefficients 13
Activity coefficients
- log[a + aAc-
H3O ] = - log Ka + log
aHAc
* activity coefficient of the undissociated acid γHAc is
essentially 1 and may be dropped.

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

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pH 에 영향 주는 인자

1. Altering the ionic strength

① Addition of neutral salts
② 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 indicator의 경우

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

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pH indicator
• Characteristics of colorimetric method

① less accurate
② less convenient but less expensive than
electrometric method
③ difficult to apply for the unbuffered
pharmaceutical preparation (change the pH -
indicator itself is acids or base)
④ error may be introduced by the presence of
salts & proteins
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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

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

ΔB : small increment in gram equivalents/Liter

of strong(or acid) added to the buffer
soln. to produce a pH change of ΔpH
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Buffer capacity (근사식 이용)
HAc + NaOH NaAC + H2 O
(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
β= = = 0.11
pH 0.09
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Buffer capacity
• A more exact equation for buffer capacity (1914, 1922)
Ka • [H3O+]
β = 2.3 • C •
(Ka + [H3O+])2

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

the acid & the salt)
.
β ---- at any [H3O+]

22
Maximum Buffer capacity

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

2
[H3O+] 2.303
βmax = 2.303 • C • = •C
+
(2 [H3O ]) 2 4

βmax = 0.576 • C
( pH = pKa )

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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(βmax) 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)

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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
① Primary buffers : Plasma ;
NaHCO3-- H2CO3, NaHPO4--NaH2PO4, protein
② 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 normal…hydrogen ions are excreted by the kidney.
- above pH 7.4…hydrogen 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
① Select a weak acid having a pKa approximately equal
to the pH at which the buffer is to be used.

② Calculate the ratio of salt & weak acid required to

obtain the desired pH.

③ 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
④ 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.

⑤ 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 – β , Volume

(b) Physiologic fluid - β , Volume

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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.
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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 value…the 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 ºC (Freezing point lowering of

human blood & lacrimal fluid)
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Calculating Tonicity Using Liso values

• The Van’t 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°
① Cryoscopic method
② Sodium chloride equivalent method

Class II …add Water to form an isotonic soln.

① White-Vincent method
② Sprowls method

44
Class I methods
• Cryoscopic method (빙점강하도법)
(Example)
How much NaCl is required to render 100mL of a 1% soln. of
apomorphine HCl isotonic with blood serum?
Δ Tf0.9% of NaCl soln : 0.52°(Isotonic with blood)
Δ Tf1% of apomorphine HCl soln : 0.08° (from table)

to reduce the freezing point by an additional 0.44°(0.52-0.08)

Δ 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
(염화나트륨당량법) by Mellen & Seltzer
1g drug tonicity = Eg NaCl tonicity
E : weight of NaCl with the same freezing point
depression as 1g of the drug.

ΔTf = Liso · c c=1g/

molecular weight
ΔTf = 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
① 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)

② 0.9 g/100mL = 0.063 g / V

③ V = 0.063 • 100/0.9
④ V = 7.0 mL
⑤ 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.

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Class II methods
• Sprowls method

w E 0.9 g
=
V 100 ml

W = 0.3 g
(1% solution)
TABLE

?
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Thank you for your attention

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