CHEMICAL FOUNDATION OF BIOCHEMISTRY - 1

Course Code: 21BCH2T411

UNIT-3
BUFFER AND COLLEGATIVE PROPERTIES

Prepared by
Dr. Austin Richard S
Department of Biochemistry
Kristu Jayanti College
Kothanur, Bengaluru - 560077
Acid
➢ The term "acid" originates from the Latin words "acidus" or "acere," which mean
"sour."
➢ The characteristics of acids is their sour taste.
➢ An acid is a substance that produces ionizable hydronium ions (H₃O⁺) in an aqueous
solution and turns blue litmus paper red. In solution, acids dissociate to form their
constituent ions, as illustrated by the following examples.

HCl (aq) H+ + Cl- or HCl + H2O → H3O+ + Cl–

H2SO4 (aq) 2H+ + SO42-
CH3COOH(aq) H+ + CH3COO -
Acids are classified into two types based on their source:
1. Natural acids
2. Mineral acids

1. Natural Acids:
These acids are derived from natural sources such as fruits and animal products.
Examples: include lactic acid, citric acid, and tartaric acid.

2. Mineral Acids:
Mineral acids are synthesized from minerals.
Examples: Hydrochloric acid (HCl), Sulfuric acid (H₂SO₄), and Nitric acid (HNO₃).
Bases
➢ The most common characteristics of bases are their bitter taste and slippery, soapy feel.
➢ A base is a substance that produces hydroxide ions (OH⁻) in an aqueous solution and
turns red litmus paper blue.
➢ In aqueous solutions, bases dissociate to release their constituent ions, as shown in the
following examples.

NaOH Na+ + OH-

Ca(OH)2 Ca2+ + 2 OH-
The Arrhenius theory
➢ The Arrhenius theory explains the behavior of acids and bases in aqueous solutions.
➢ The theory states that
1. An acid is a substance that releases hydrogen ions (H⁺) in an aqueous solution.
2. A base releases hydroxyl ions (OH⁻) in an aqueous solution.
3. In a aqueous solution, a hydrogen ion does not exist independently; instead, it
combines with a water molecule to form a hydronium ion (H₃O⁺)

Arrhenious Acid: HCl (aq) → H+ (aq) + Cl– (aq) or HCl + H2O → H3O+ + Cl–

Arrhenious Base: NaOH (aq.) → Na+ (aq.) + OH– (aq.)

Acid-base reactions: Arrhenius acid + Arrhenius base = water + salt
When an Arrhenius acid reacts with an Arrhenius base, the products are typically
water and a salt. These reactions are commonly referred to as neutralization
reactions.

Example :
The reaction between Hydrogen fluoride (HF) and lithium hydroxide (LiOH) results in lithium fluoride
(LiF) and water (H2O).
HF (aq.) + LiOH (aq.) → LiF (aq) + H2O
Let us look at the reaction step-by-step.
Step1 : HF (aq.) → H+ (aq.) + F– (aq.)
Step2 : LiOH (aq.) → Li+ (aq.) + OH– (aq.)
Li+ and F– combine to form LiF, which is a salt. H+ and OH– react to form water.
The overall reaction is given by
HF (aq.) + LiOH (aq.) → LiF (aq.) + H2O
Limitation of The Arrhenius theory

1. The Arrhenius theory only applies to acids and bases in aqueous solutions, but acid-
base reactions can also occur in gaseous phases. The theory fails to explain such
reactions, which is explained the Brønsted-Lowry theory.

1. According to the Arrhenius theory, the presence of H⁺ and OH⁻ ions is essential in
chemical reactions. However, compounds like aqueous sodium bisulfate (NaHSO₄) and
sodium carbonate (Na₂CO₃) do not fit this theory. NaHSO₄ is acidic despite lacking H⁺
ions, and Na₂CO₃ is basic despite not containing OH⁻ ions.
 Lowry and Bronsted concepts
According to Bronsted-Lowry theory,
➢ Acid is a substance which donates a proton or H+ ion to the other compound and forms a
conjugated base.
➢ Base is a substance which accepts a proton or H+ ion from the other compound and forms
conjugated acid.
Example :
For example , consider the following chemical equation:
HCl(aq) + NH3(aq) → NH4+ (aq)+ Cl−(aq)
Here, hydrochloric acid (HCl) "donates" a proton (H+) to ammonia (NH3) which "accepts" it , forming a positively charged
ammonium ion (NH4+) and a negatively charged chloride ion (Cl-).
Therefore, HCl is a Brønsted-Lowry acid (donates a proton) while the ammonia is a Brønsted-Lowry base (accepts a
proton). Also, Cl- is called the conjugate base of the acid HCl and NH4+ is called the conjugate acid of the base NH3
 LEWIS’S CONCEPT OF ACID AND BASES
▪ Lewis theory of acid-base reactions defines:
▪ Lewis acids as electron-pair acceptors.
▪ Lewis bases as electron-pair donors.
▪ A Lewis base donates a pair of electrons to a Lewis acid. This leads to the
formation of a coordinate covalent bond between the acid and base. The
product formed from this reaction is called a Lewis adduct.
 LEWIS ACIDS
➢ Lewis acids are chemical species with empty orbitals that can accept
electron pairs from Lewis bases.
➢ These species typically have a trigonal planar structure and an empty p-
orbital.
➢ An example of a Lewis acid is BF₃, AlCl3 .
➢ Water and certain other compounds can act as both Lewis acids and bases
because they can either accept or donate electron pairs depending on the
reaction.
 LEWIS BASES
➢ Lewis bases are atomic or molecular species with a highly localized HOMO (Highest
Occupied Molecular Orbital: molecular orbital that contains the highest energy electrons in a
molecule).
➢ These species have the ability to donate an electron pair to a Lewis acid, forming an adduct.
➢ Common examples of Lewis bases include ammonia, alkyl amines, and other conventional
amines.
➢ Most Lewis bases are anionic in nature.
➢ Being electron-rich species, Lewis bases can donate electron pairs, making them
nucleophiles.
Examples :
➢ Pyridine and its derivatives can act as electron pair donors, making them Lewis bases.
➢ Compounds where Oxygen, Sulfur, Selenium, and Tellurium (elements from group 16
of the Periodic Table) exhibit an oxidation state of -2 are typically Lewis bases.
➢ Examples include water and ketones.
➢ Simple anions with electron pairs, such as H⁻ and F⁻, can function as Lewis bases by
donating these electrons.
➢ Even complex anions, like the sulfate anion (SO₄²⁻), can donate electron pairs.
➢ π-systems rich in electrons, such as benzene, ethyne, and ethene, also have strong
electron pair donating capabilities.
The H⁺ ion functions as a Lewis acid, while H₂O serves as a Lewis base. When the water molecule reacts with the proton, it
forms a hydronium ion (H₃O⁺), as shown in the illustration below. Here, the oxygen atom donates an electron pair to the
proton, forming a coordinate covalent bond in the process.

The proton accepts an electron pair from the nitrogen atom of the ammonia molecule. This leads to the formation of a
coordinate covalent bond between them, resulting in the creation of a Lewis adduct (the ammonium cation).

Ammonia acts as a Lewis base and the silver ion acts as a Lewis acid. Each nitrogen atom donates an electron pair to
Ag+,
 BUFFERS

Buffer is a solution whose pH is not altered to any great extent by the addition
of small quantities of either an acid or base.

Many chemical reactions take place at a constant pH.

In nature, numerous systems rely on buffering mechanisms to regulate pH

levels. For example, the bicarbonate buffering system helps maintain the pH of
blood.
Types of buffer
(a)Acidic Buffer:
Formed by mixing a weak acid with its salt derived from a strong base.
Examples : CH₃COOH + CH₃COONa

CH3COOH ⇋ CH3COO– + H+ (incomplete dissociation)

CH3COONa → CH3COO– + Na+ (complete dissociation)
H2O ⇋ H+ + OH– (incomplete dissociation)

Action of acid: When a drop of stong acid (HCl) is added in the above buffer solution H+ ions
combine with CH3COO- ions to form weakly ionized CH3COOH. Whose ionization is further
suppressed due to common ion effect. So pH of the solution unaltered.

Action of base: when a drop of strong base (NaOH) is added to the above buffer solution it
react with free acid to form water molecules. So pH of the solution unaltered.
CH3COOH + OH– ⇋ CH3COO– + H2O
(b) Basic Buffer
Formed by mixing a weak base with its salt derived from a strong acid.
It is the mixture of NH4OH and NH4Cl in aqueous solution(Ammonium buffer).

NH4OH ⇋ NH4+ + OH– (incomplete dissociation)

NH4Cl → NH4+ + Cl– (complete dissociation)

H2O ⇋ H+ + OH– (incomplete dissociation)

Action of acid: when a drop of HCl is added, the added H+ ions combine with NH4OH to form
water molecules. So the pH of buffer is unaffected.

NH4OH + H+ ⇋ NH4+ + H2O

Action of base: when a drop of NaOH is added, the added OH– ions combine with NH4+ ions
to form weakly ionized NH4OH. It is further suppressed due to common ion effect. So the pH
of buffer is unaffected.
pH
➢ pH is defined as the negative logarithm of H+ ion concentration.
➢ pH can determine the strength of acids and bases quantitatively.

pH scale
➢ A pH scale is a tool for measuring acids and bases. The scale ranges from 0 - 14.
➢ The pH is the concentration of the hydrogen ions. This calculation is based on a
pH scale.
➢ The pH scale is logarithmic and shows the solution’s concentration of hydrogen ions
inversely.

➢ More specifically, the pH of the solution is the negative logarithm to the base 10 of its
hydrogen ion concentration in moles per litre.

pH = - log [H+] or pH = log [1/H+]

Why Logarithm in pH ?

➢ The concentration of hydrogen ions can vary across many orders of magnitude — from 1
to 0.00000000000001 moles per liter.

➢ A change of one pH unit corresponds to a ten-fold change in hydrogen ion concentration

 pKa
The pKa value is the negative base -10 logarithm of the acid dissociation constant (Ka) of a
solution.
pKa = - log 10 Ka

➢ pKa describes the acidity of a particular molecule.

➢ It measures the strength of an acid

➢ The lower the value of pKa, the stronger the acid and the greater its ability to donate its

protons.

➢ Ka denotes the acid dissociation constant. It measures how completely an acid dissociates in

an aqueous solution. The larger the value of Ka, the stronger the acid as acid largely

dissociates into its ions and has lower pka value.

 Henderson-Hasselbalch equation

➢ Describe the derivation of pH as measure of acidity in biological sample

➢ The equation is useful to estimation of pH in a buffer solution

The Henderson-Hasselbalch equation can be written as:

pH = pKa + log10 ([A–]/[HA])

[A–] denotes the molar concentration of the conjugate base (of the acid)
[HA] denotes the molar concentration of the weak acid.

According to Bronsted –Lowry theory Acid is the Substance which donate the proton
Base is the substance which accept the he proton
 Derivation of Henderson-Hasselbalch equation
Let us take an example of ionization of weak acid HA :

HA + H2O ⇌ H+ + A−
(weak acid) (conjugate base)
Acid dissociation constant, Ka can be given as:

Ka​ = [H+][A−]​
[HA]

Taking, - log 10 of RHS and LHS:

- log10 Ka = - log10 [H+][A−]​ (log10 mn = log10m + log10n)

[HA]

- log10 ka = - log10 [H+] - log10 [A −]

[HA]

pKa = pH - log10 [A −]
[HA]

pH = pKa + log10 [A −]
[HA]
 Derivation of Henderson-Hasselbalch equation
Let us take an example of ionization of weak Base HA :

B + H2O ⇌ OH- + HB+

(conjugate acid)
Acid dissociation constant, Kb can be given as:

Kb​ = [OH-][HB+]​
[B]

Taking, - log 10 of RHS and LHS:

- log10 Kb = - log10 [OH-][HB+] (log10 mn = log10m + log10n)

[B]

- log10 Kb = - log10 [OH-] - log10 [HB+]

[B]

pKb = pOH - log10 [HB+]

[B]

pOH = pKb + log10 [HB+]

[B]
Titration curve of phosphoric acid H3PO4

At pH 9.6, indicator
thymolphthalein
turns to blue

At pH 4.7 methyl orange turn to yellow

 Colligative properties
The term "colligative" is derived from the Latin word "colligatus," meaning "bound together."

In the context of solutions, colligative properties help explain how the properties of a

solution are connected to the concentration of solute present in it.

➢ Colligative properties depend on the ratio of solute particles to solvent particles in a

solution.

➢ They do not depend on the chemical nature of the solute or solvent.

➢ Colligative properties are related to the concentration of a solution, expressed by

molarity, normality, or molality.

 Colligative properties

➢ The four colligative properties exhibited by a solution are:

1. Boiling point elevation
2. Freezing point depression
3. Relative lowering of vapor pressure
4. Osmotic pressure.

➢ All these properties depend on the number of solute particles irrespective of their
nature relative to the total number of particles present in the solution. Such properties
are called colligative properties (colligative: from Latin: co means together, ligare
means to bind).
 Vapour pressure
➢ Evaporation is a type of vaporization that occurs on the surface of a liquid as it changes into
the gas phase.
➢ Vapour pressure is defined as the pressure exerted by the vapour present above the liquid.
➢ Vapor pressure or equilibrium vapor pressure is the pressure of a vapor in thermodynamic
equilibrium with its condensed phases in a closed container.
ELEVATION OF BOILING POINT

The vapour pressure of a liquid increases with increase of temperature. It boils at the
temperature at which its vapour pressure is equal to the atmospheric pressure.

For example water boils at 373.15 K (100° C) because at this temperature the vapour
pressure of water is 1.013 bar (1 atmosphere)
 ➢ The boiling point of a solution is always higher than
that of the boiling point of the pure solvent in which
the solution.

➢ lowering of vapour pressure, the elevation of boiling

point also depends on the number of solute
molecules rather than their nature.

ΔTb = Tb- T0
ΔTb = Elevation in the boiling Point
Tb = Boiling point of Solution
Figure : The vapour pressure curve T0 = Boiling point of Pure solvent
for solution lies below the curve
for pure water.
Solutions the elevation of boiling point (ΔTb) is directly proportional to the molal
concentration of the solute in a solution
ΔTb = Boiling point elevation
ΔTb α m Kb = Boiling point elevation constant
ΔTb = Kb m M= Molality
ΔTb = Boiling point elevation
Kb = Boiling point elevation constant
ΔTb = Kb x w2 /M2 w2= weight of the solute in g
M2= Molecular weight of the solute
w1 /1000 W1 = weight of the Solvent in g

ΔTb = Kb x w2 x 1000
M2 x w1
 Freezing point depression
➢ The lowering of vapour pressure of a solution causes a lowering of the freezing point
compared to that of the pure solvent
➢ The freezing point is the temperature where the vapor pressure of a substance in its
liquid phase equals the vapor pressure in its solid phase.
➢ According to Raoult’s law, adding a non-volatile solute to a solvent lowers its vapor
pressure.
➢ As a result, the vapor pressure of the solution matches that of the solid solvent at a
lower temperature.
➢ The difference between the freezing point of the pure solvent and the solution is known
as the depression in freezing point.
➢ A solution will freeze when its vapour pressure equals the
➢ vapour pressure of the pure solid solvent.
➢ According to Raoult’s law, when a non-volatile solid is added
to the solvent its vapour pressure decreases and now it
would become equal to that of solid solvent at lower
temperature.
Depression in the freezing point
ΔTf = Tf0- Tf
ΔTb = Depression of freezing point
Tf0 = Freezing point of pure solvent
Tf = Freezing point of Solution
Depression of freezing point for dilute Solution (ΔTf) is directly proportional to the molal
concentration of the solute in a solution

ΔTf α m ΔTb = Depression in the freezing point

ΔTf = Kf m Kf = Freezing point depression constant

ΔTf = Kf x w2 /M2 M = Molality

w1 /1000
ΔTf = Kf x w2 x 1000
M2 x w1 ΔTb = Depression in the freezing point
Kf = Freezing point depression constant
w2= weight of the solute in g
M2= Molecular weight of the solute
W1 = weight of the Solvent in g
Relative lowering of vapor pressure
Let p1 the vapor pressure of the solvent in a solution, x1 be its mole fraction of solvent in a
solution, P10 be the vapor pressure of pure solvent.

Then according to Raoult’s law “In a binary solution the partial vapor pressure of each
volatile component in the solution is directly proportional to its mole fraction ”.
p1 α x 1
p1 = x1 p10 --------> eq. 1
Reduction in vapour pressure can be represent as
Δ p = p10 - p1 (apply eq. 1 for p1 )
= p10 - x1 p10
= p10 (1- x1) --------> eq. 2

Knowing that x2 = 1 – x1, (apply x2 value in eq. 2 )

Δ p = p10 x2 x2 = Δp
p10
Δ p = p10 x2

x2 = Δp
p10
Here n1 and n2 are the number of moles of solvent
and solute respectively present in the solution.

n2 Δp For dilute solutions n2<<n1, hence neglecting n2 in

n1+n2
=
p10 the denominator we have

n1 = number of moles of solvent

n2 = number of moles of solute
n2 Δp
= Δ p = reduction of vapour pressure
n1 p10
P10 = Partial vapour pressure of pure solvent
 Osmotic Pressure
Osmosis
Osmosis is the movement of solvent from a region of lower solute concentration to a region of
higher solute concentration through a semi-permeable membrane. The flow will continue till
the equilibrium is attained.

Osmotic Pressure
Osmotic pressure can be defined as the
minimum pressure that must be applied to a
solution to just stop the flow of solvent
molecules through a semipermeable
membrane.
The formula for osmotic pressure is π = iCRT,
where:
π = Osmotic pressure
i = Van’t Hoff factor
C = Molar concentration of the solute in the solution
R = Universal gas constant
T = Temperature
 The Valence Shell Electron Pair Repulsion Theory (VSEPR)

LONE PAIR ELECTRONS

A lone pair electrons refers to a pair of valence electrons which do not participate in
chemical bonding with another atom, It is also called a non-bonding pair.

BOND PAIR ELECTRONS

A bond pair electrons refers to a pair
of valence electrons which
participate in chemical bonding
with another atom.
Geometry of the Molecule
Geometry is determined by the both the bond pair and lone pair of electrons.

Geometry : Tetrahedron
Shape of the Molecule
Shape is determined by the bond pair of electrons.

Shape : Bent
Main Points of VSEPR THEORY

1. Electron pair repel each other and tends to stay away .

2. Different electron pair have different types of repulsion.

Lone pair : Lone Pair > Lone pair: bond pair > Bond pair :Bond pair

3. Geometry and shapes is determined by bond pairs and lone pairs.

a. If lone pair is Zero in a molecule, Geometry = shape.
b. If lone pair is not Zero in a molecule, Geometry ≠ shape.
 2 atom bonding with central atom

1. BeCl2
Bond pair :2
Lone pair :0
Geometry : Linear
Shape : Linear 180 ⁰
Hybridization : sp
Bond angle : 180⁰
Shape = Geometry
 2 atom bonding with central atom
2. H2O
Bond pair :2
Lone pair :2
Geometry : Tetrahedral
Shape : Bent
Hybridization : sp3
Geometry of water molecule
Bond angle : 105⁰
Shape ≠ Geometry

Shape of water molecule

2. CO2
Bond pair :4
Lone pair :0
Geometry : Linear
Shape : Linear
Hybridization : sp 180 ⁰
Bond angle :180⁰
Shape = Geometry
 3 atoms bonding with central atom

1. BF3
Bond pair :3
Lone pair :0
Geometry : Triogonal
Shape : Triogonal
Hybridization : sp2
Bond angle : 120⁰
Shape = Geometry
2. SO2
Bond pair :4
116⁰
Lone pair :1
Geometry : Triogonal Lone pair: bond pair > Bond pair :Bond pair

Shape : Bent
Hybridization : sp2
Bond angle : 116⁰
Shape ≠ Geometry
4 atoms bonding with central atom
Ionic product of water:
Ionic product of water is defined as the product of molar concentrations of H+ and OH-at a
specified temperature.

The ionization of water can be represented as:

H2O(l) H+ (aq) + OH- (aq)
Effect of temperature
The ionization constant is given
as shown below. 1. With the rise in temperature, there is increase in
concentrations of these ions and hence the value of the
K = [H+][OH-] ionic product also increases.
[H2O]
2. At 25°C, the value of ionic product is 1×10-14 mol2 /L2

K [H2O] = [H+] [OH-] 3. Therefore, H+= OH- =1×10-7 mol/ltr.

4. But for different temperatures, these values may be
Kw = [H+] [OH-] different and can be calculated accordingly.

@ 25 ⁰C Kw = [10-7mol /L] [10-7mol /L] = 10-14 mol2 /L2

 BUFFERS IN ANIMAL SYSTEM
Acid Base Balance

➢ The normal pH of the blood is 7.35-7.45

➢ Erythrocytes pH is 7.2 (intracellular pH).

➢ Changes in blood pH will alter the intracellular pH which, in turn, influence

the metabolism e.g. distortion in protein structure, enzyme activity etc.

➢ It is estimated that the blood pH compatible to life is 6.8-7.8

Production of acids by the body
➢ Carbonic acid, lactic acid, sulfuric acid, phosphoric acid etc.
➢ A diet rich in animal proteins results in more acid production by the
body.

Production of bases by the body

➢ The formation of basic compounds in the body is negligible
➢ Bicarbonates, Ammonia.
 Maintenance of Blood pH

Blood pH is maintained by three mechanism

1. Blood buffers

2. Respiratory mechanism

3. Renal mechanism.
1. BLOOD BUFFERS

Buffers are solutions which can resist changes in

pH when small amount of acid or alkali is
added.

The blood contains 3 buffer systems.

1. Bicarbonate buffer
2. Phosphate buffer
3. Protein buffer.
1. Bicarbonate buffer

➢Sodium bicarbonate & carbonic acid (ratio 20:1)

(NaHCO3- H2CO3).

➢Predominant buffer of the extracellular fluid

➢Referred as Alkali reserve of the in the body.

2. Phosphate buffer system
➢ Sodium dihydrogen phosphate & disodium hydrogen phosphate

➢ ( NaH2PO4- Na2HPO4) - Ratio 4:1

➢ pKa of 6.8 (closet o blood pH 7.4)

➢ Intracellular buffer and low concentration in plasma.

3. Protein Buffer
➢ Constitute Plasma proteins & hemoglobin .
➢ Account for about 2% of the total buffering capacity of the plasma.
➢ The buffering capacity is dependent on the pKa of the ionizable groups of
amino acids.
➢ The imidazole group of histidine (pK = 6.7) is the most effective
contributor of protein buffers.
➢ Hemoglobin buffers the fixed acids.
 Boyles Law
➢ Formulated by: Anglo-Irish chemist Robert Boyle in
1662.
➢ Stating the relationship between pressure and volume
of a gas.
➢ It says that pressure exerted by a gas (at constant
amount (mole) and constant temperature) is inversely
proportional to the volume it occupies.
➢ As pressure increases, volume decreases, and vice
versa, provided the temperature and quantity of gas
remain constant.
For a gas, the relationship between volume and
pressure (at constant mass and temperature) can be
expressed mathematically as follows.

P ∝ (1/V)

Where P is the pressure exerted by the gas and V is

the volume occupied by it. This proportionality can be
converted into an equation by adding a constant, k.

P = k*(1/V) ⇒ PV = k
In mathematical terms, this relationship is expressed as

P1V1 = P2V2,

P1 : initial pressure,

P2 : final pressure,

V1 : initial volume and

V2 : final volume.

This equation highlights the inverse relationship between pressure and volume. As one

variable increases, the other decreases, and vice versa, assuming temperature remains

constant.
Charles' Law
➢ Formulated by French physicist Jacques Charles(1780)
➢ States that the volume(V) of an ideal gas is directly
proportional to its absolute temperature (T in Kelvin)
at constant pressure.
➢ When the pressure on a sample of dry gas is kept
constant, the temperature and volume are in direct
proportion.
V1 V2
=
T1 T2
𝑉∝𝑇 V / T = K(constant)
Mathematically:
𝑉∝𝑇 V / T = K(constant)
As temperature increases, volume increases, and
vice versa.
Determination of osmotic pressure: Berkeley and Hartley’s method
Berkeley and Hartley’s method employed the technique of applying external pressure on the
solution just enough to prevent osmosis (Osmotic pressure).

Apparatus
A porcelain tube having Cu2Fe(CN)6 copper
ferrocyanide membrane in its walls in enclosed in
a metallic jacket. The porcelain tube is fitted with
a reservoir of pure solvent at one end and a
capillary tube at the other end. In a metallic
jacket is an arrangement for applying external
pressure which is measured with the help of
pressure gauze.
Procedure:
➢ The porcelain tube is filled with pure solvent and the
metallic jacket with solution.
➢ The level in the capillary tube will tend to move
down as the solvent flows towards solution due to
osmosis.
➢ External pressure is now applied on the solution by
the piston so that level in capillary remain stationary.
The reading of pressure gauge is recorded. This is the
osmotic pressure of the solution.
Advantages:
➢ Quick and Accuracy : It is a quick and accurate method to measure
osmotic pressure.
➢ Reliable :The concentration of the solution does not change because flow
of solvent is not permitted into solution;
➢ Good Design : As the osmotic pressure is balanced by external pressure,
there is no strain on membrane and the danger of its bursting is
eliminated.
➢ Method can be used to measure high osmotic pressure