PHYSICOCHEMICAL

ASPECTS
• Water is the most abundant
substance in living systems, making
up 70% or more of the weight of
most organisms
• The attractive forces between water
molecules and the slight tendency of
water to ionize are of crucial
importance to the structure and
function of biomolecules
• The water molecule and its
ionization products, H and OH ,
+ -

profoundly influence the

–structure,
–self-assembly,
–properties of all cellular
components, including proteins,
nucleic acids, and lipids
Hydrogen Bonding gives Water its Unusual
Properties
• Water has a higher melting point, boiling point,
and heat of vaporization than most other
common solvents
• These unusual properties are a consequence of
attractions between adjacent water molecules
that give liquid water great internal cohesion
• A look at the electron structure of the H2O
molecule reveals the cause of these
intermolecular attractions
• Each hydrogen atom of a water
molecule shares an electron pair
with the central oxygen atom

• The geometry of the molecule is

dictated by the shapes of the
outer electron orbitals of the
oxygen atom
• These orbitals describe a
rough tetrahedron, with
a hydrogen atom at each
of two corners and
unshared electron pairs
at the other two corners
• The oxygen nucleus
attracts electrons more
strongly than does the
hydrogen nucleus
• That is, oxygen is more
electronegative
• This means that the
shared electrons are
more often in the
vicinity of the oxygen
atom than of the
hydrogen
• The result of this
unequal electron
sharing is two electric
dipoles in the water
molecule, one along
each of the H-O bonds
• Each hydrogen bears a
partial positive charge (δ+ ),
and the oxygen atom bears
a partial negative charge
equal in magnitude to the
sum of the two partial
positives (2δ+ )
• As a result, there is an
electrostatic attraction
between the oxygen atom
of one water molecule and
the hydrogen of another,
called a hydrogen bond
• Hydrogen bonds are relatively weak
• Those in liquid water have a bond
dissociation energy (the energy re
required to break a bond) of about 23
kJ/mol, compared with 470 kJ/mol for
the covalent O-H bond in water or 348
kJ/mol for a covalent C-C bond
• The sum of all the hydrogen bonds
between H2O molecules confers great
internal cohesion on liquid water
• At room temperature, the thermal
energy of an aqueous solution (the
kinetic energy of motion of the
individual atoms and molecules) is of
the same order of magnitude as that
required to break hydrogen bonds
• When water is heated, the increase in
temperature reflects the faster
motion of individual water molecules
• The nearly tetrahedral arrangement of the
orbitals about the oxygen atom allows each
water molecule to form hydrogen bonds
with as many as four neighboring water
molecules
• In liquid water at room temperature and
atmospheric pressure, however, water
molecules are disorganized and in
continuous motion, so that each molecule
forms hydrogen bonds with an average of
only 3.4 other molecules
 Water Forms Hydrogen Bonds With Polar Solutes
• Hydrogen bonds are not
unique to water
• They readily form between
an electronegative atom
(the hydrogen acceptor,
usually oxygen or nitrogen)
and a hydrogen atom
covalently bonded to
another electronegative
atom (the hydrogen donor)
in the same or another
molecule
Water Interacts Electrostaticaily With
Charged Solutes
• Water is a polar solvent
• It readily dissolves most
biomolecules, which are generally
charged or polar compounds
• Compounds that dissolve easily in
water are hydrophilic (Greek, "water-
loving")
Nonpolar Compounds Force Energeticaly
favorable Changes in the Structure of Water
• When water is mixed with benzene or hexane,
two phases form; neither liquid is soluble in
the other
• Nonpolar compounds such as benzene and
hexane are hydrophobic-they are unable to
undergo energetically favorable interactions
with water molecules, and they interfere with
the hydrogen bonding among water
molecules
• All molecules or ions in aqueous solution
interfere with the hydrogen bonding of
some water molecules in their immediate
vicinity, but polar or charged solutes
(such as NaCl) compensate for lost water-
water hydrogen bonds by forming new
solute-water interactions
• The net change in enthalpy (energy
change, H) for dissolving these solutes is
generally small
• Hydrophobic solutes, however, offer
no such compensation, and their
addition to water may therefore
result in a small gain of enthalpy
• The breaking of hydrogen bonds
between water molecules takes up
energy from the system, requiring
the input of energy from the
surroundings
• In addition to requiring this input of
energy, dissolving hydrophobic
compounds in water produces a
measurable decrease in entropy
• Water molecules in the immediate
vicinity of a nonpolar solute are
constrained in their possible
orientations as they form a highly
ordered cagelike shell around each
solute molecule
• The number of ordered water
molecules, and therefore the
magnitude of the entropy decrease,
is proportional to the surface area of
the hydrophobic solute enclosed
within the cage of water molecules
• The free-energy change for dissolving
a nonpolar solute in water is thus
unfavorable
Solutes Affect the Colligative
Properties of Aqueous Solutions
• Solutes of all kinds alter certain
physical properties of the solvent,
water:
–vapor pressure,
–boiling point,
–melting point (freezing point),
–osmotic pressure
• These are called colligative
properties (colligative meaning"
tied together"), because the effect
of solutes on all four properties
has the same basis:
–the concentration of water is
lower in solutions than in pure
water
• The effect of solute concentration
on the colligative properties of
water is independent of the
chemical properties of the solute

• It depends only on the number of

solute particles (molecules, ions)
in a given amount of water
• A compound such as NaCl, which
dissociates in solution, has an effect on
osmotic pressure, for example, that is
twice that of an equal number of moles
of a nondissociating solute such as
glucose
• Water molecules tend to move from a
region of higher water concentration to
one of lower water concentration, in
accordance with the tendency in nature
for a system to become disordered
• When two different aqueous
solutions are separated by a
semipermeable membrane (one that
allows the passage of water but not
solute molecules), water molecules
diffusing from the region of higher
water concentration to the region of
lower water concentration produce
osmotic pressure
• Osmosis, water movement across a
semipermeable membrane driven
by differences in osmotic pressure,
is an important factor in the life of
most cells
• Plasma membranes are more
permeable to water than to most
other small molecules, ions, and
macromolecules
• This permeability is due
largely to protein channels
(aquaporins) in the
membrane that selectively
permit the passage of water
• Solutions of
osmolarity equal
to that of a cell's
cytosol are said to
be isotonic relative
to that cell
• Surrounded by an
isotonic solution, a
cell neither gains
nor loses water
• In a hypertonic
solution, one
with higher
osmolarity than
that of the
cytosol, the cell
shrinks as
water moves
out
• In a hypotonic solution,
one with a lower
osmolarity than the
cytosol, the cell swells as
water enters
• In their natural
environments, cells
generally contain higher
concentrations of
biomolecules and ions
than their surroundings,
so osmotic pressure
tends to drive water into
• If not somehow counterbalanced, this
inward movement of water would distend
the plasma membrane and eventually cause
bursting of the cell (osmotic lysis)
• Several mechanisms have evolved to
prevent this catastrophe
• In bacteria and plants, the plasma
membrane is surrounded by a
nonexpandable cell wall of sufficient rigidity
and strength to resist osmotic pressure and
prevent osmotic lysis
• Certain freshwater micro organisms
that live in a highly hypotonic
medium have an organelle
(contractile vacuole) that pumps
water out of the cell
• In multicellular animals, blood plasma
and interstitial fluid are maintained at
an osmolarity close to that of the
cytosol
• The high concentration of
albumin and other proteins in
blood plasma contributes to its
osmolarity
• Cells also actively pump out Na+
and other ions into the interstitial
fluid to stay in osmotic balance
with their surroundings
• Because the effect of solutes on osmolarity
depends on the number of dissolved particles,
not their mass, macromolecules (proteins,
nucleic acids, polysaccharides) have far less
effect on the osmolarity of a solution than
would an equal mass of their monomeric
components
• Storing fuel as polysaccharides (starch or
glycogen) rather than as glucose or other
simple sugars avoids an enormous increase in
osmotic pressure in the storage cell
TO SUMMARIZE
• The very different electronegativities of H and O
make water a highly polar molecule, capable of
forming hydrogen bonds with itself and with
solutes
• Hydrogen bonds are fleeting, primarily
electrostatic, and weaker than covalent bonds
• Water is a good solvent for polar (hydrophilic)
solutes, with which it forms hydrogen bonds, and
for charged solutes, with which it interacts
electrostatically
• Nonpolar
(hydrophobic)
compounds dissolve
poorly in water; they
cannot hydrogen-
bond with the solvent,
and their presence
forces an energetically
unfavorable ordering
of water molecules at
their hydrophobic
surfaces
• To minimize the surface
exposed to water,
nonpolar compounds
such as lipids form
aggregates (micelles) in
which the hydrophobic
moieties are
sequestered in the
interior, associating
through hydrophobic
interactions, and only
the more polar
moieties interact with
water
• The physical properties of aqueous solutions are
strongly influenced by the concentrations of
solutes
• When two aqueous compartments are separated
by a semipermeable membrane (such as the
plasma membrane separating a cell from its
surroundings), water moves across that
membrane to equalize the osmolarity in the two
compartments
• This tendency for water to move across a
semipermeable membrane is the osmotic
pressure
Ionization of Water, Weak Acids, and Weak
Bases
• Although many of the solvent properties of
water can be explained in terms of the
uncharged H2O molecule, the small degree of
ionization of water to hydrogen ions (H+) and
hydroxide ions (OH-) must also be taken into
account
• Like all reversible reactions, the ionization of
water can be described by an equilibrium
constant
• When weak acids are dissolved in
water, they contribute H by
+

ionizing; weak bases consume H+

by becoming protonated

• These processes are also governed

by equilibrium constants
• The total hydrogen ion concentration
from all sources is experimentally
measurable and is expressed as the
pH of the solution
• To predict the state of ionization of
solutes in water, we must take into
account the relevant equilibrium
constants for each ionization reaction
Pure Water ls Slightly lonized
• Water molecules have a slight tendency to
undergo reversible ionization to yield a
hydrogen ion (a proton) and a hydroxide ion,
giving the equilibrium reaction
H2O  H+ + OH-

• Although we commonly show the dissociation

product of water as H+, free protons do not
exist in solution
• Hydrogen ions formed in water are
immediately hydrated to
hydronium ions (H3O+)

• Hydrogen bonding between water

molecules makes the hydration of
dissociating protons virtually
instantaneous
• The ionization of water can be measured
by its electrical conductivity
• Pure water carries electrical current as
H3O+ migrates toward the cathode and
OH- toward the anode
• The movement of hydronium and
hydroxide ions in the electric field is
extremely fast compared with that of
other ions such as Na+, K+, and Cl-
• This high ionic mobility
results from the kind of
"proton hopping“ shown
• No individual proton
moves very far through
the bulk solution, but a
series of proton hops
between hydrogen-
bonded water molecules
causes the net
movement of a proton
over a long distance in a
remarkably short time
• As a result of the high ionic mobility
of H (and of OH , which also moves
+ -

rapidly by proton hopping, but in the

opposite direction), acid base
reactions in aqueous solutions are
exceptionally fast
• As noted above, proton hopping very
likely also plays a role in biological
proton-transfer reactions
• Because reversible ionization is crucial to the
role of water in cellular function, we must
have a means of expressing the extent of
ionization of water in quantitative terms
• A brief review of some properties of reversible
chemical reactions shows how this can be
done
• The position of equilibrium of any chemical
reaction is given by its equilibrium constant,
Keq (sometimes expressed simply as K )
• For the generalized reaction

A+BC+D

• An equilibrium constant can be defined in

terms of the concentrations of reactants (A
and B) and products (C and D) at
equilibrium:

Keq = [C][D]/[A][B]
• The equilibrium constant is fixed and
characteristic for any given chemical reaction
at a specified temperature
• It defines the composition of the final
equilibrium mixture, regardless of the starting
amounts of reactants and products
• Conversely, we can calculate the equilibrium
constant for a given reaction at a given
temperature if the equilibrium concentrations
of all its reactants and products are known
The lonization 0f Water ls Expressed By an
Equilibrium Constant
• The degree of ionization of water at
equilibrium is small
• At 250C only about two of every 109 molecules
in pure water are ionized at any instant
• The equilibrium constant for the reversible
ionization of water is

Keq = [H⁺][OH⁻]/[H₂O]
• In pure water at 250C, the concentration of
water is 55.5 M-grams of H2O in I L divided by
its gram molecular weight:
(1,000 g/L)/(18.015 g/mol)
• It is essentially constant in relation to the very
low concentrations of H+ and OH-, namely,1 x
10-7 M
• Accordingly, we can substitute 55.5 M in the
equilibrium constant expression to yield
Keq = [H⁺][OH⁻]/55.5
• On rearranging, this becomes
(Keq)(55.5)= [H⁺][OH⁻]= Kw

• Where Kw designates the product

(55.5M)(Keq), the ion product of water at
250C
• The value for Keq, determined by
electrical conductivity measurements of
pure water, is
1.8 X 10-16 M at 250C
• Substituting this value for Keq in the
equation gives the value of the ion
product of water
Kw =
(1.8 x 10⁻¹⁶M) (55.5M)= [H⁺]
[OH⁻]

Kw = 1.0 x 10⁻¹⁴ M²= [H⁺][OH⁻]

• Thus the product [H+][OH-] in aqueous
solutions at 250C always equals 1 x 10-14 M2
• When there are exactly equal
concentrations of H+ and OH- , as in pure
water, the solution is said to be at neutral
pH
• At this pH, the concentration of H+ and OH-
can be calculated from the ion product of
water as follows:
Kw= [H⁺][OH⁻]= [H⁺]² = [OH-]²
• Solving for[H⁺] gives:
[H⁺]= √ Kw = √ 10⁻¹⁴ M²
[H⁺]= 10⁻⁷ M

• As the ion product of water is

constant, whenever [H+] is greater
than 1 x l0-7M, [OH-] must be less
than 1 X 10 M, and vice versa
-7
• When [H+] is very high, as in a
solution of hydrochloric acid,
[OH ] must be very low
-

• From the ion product of water

we can calculate [H ] if we
+

know [OH ], and vice versa

-
W0RKED EXAMPLE- Calculation of [H+]
• What is the concentration of H+ in a solution of
0.1 M NaOH?
Solution:
• We begin with the equation for the ion product of
water:
Kw= [H⁺][OH⁻]
• With [OH-] = 0.1 M, solving for [H+] gives
• [H⁺] = Kw/[OH⁻]
= 1 × 10-14M2/ 0.1M
= 10-14M2/0.1M
= 10-13M
• What is the concentration of OH- in a solution
with an H+ concentration of 1.3x 10-4M?
Solution:
• We begin with the equation for the ion
product of water:
Kw= [H⁺][OH⁻]
With [H+] = 1.3x 10-4M, solving for [OH-] gives
[OH-] = Kw/[H+]
= 1x10-14M2/ 1.3x 10-4M
= 7.7 x 10-11
The pH Scale Designates the H+ and
OH Concentrations
-

• The pH of a solution is defined as the

logarithm to the base 10 of the
reciprocal of the [H+], ie, the negative
logarithm of the [H ]+

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

• The symbol p denotes "negative
logarithm of"
• The pH of water at 25°C, in which
H and OH ions are present in
+ –

equal numbers, is 7.0

• pH = log 1/1 x 10-7 = 7
• For each pH unit less than 7.0, the
[H ] is increased tenfold; for each
+

pH unit above 7.0, it is decreased

tenfold
• In the plasma of healthy
individuals, pH is slightly alkaline,
maintained in the narrow range of
7.35 to 7.45
• Conversely, gastric fluid pH can be
quite acidic (on the order of 2.0)
and pancreatic secretions can be
quite alkaline (on the order of 8.0)
 pH of body fluids
BODY FLUID pH
Blood plasma 7.35-7.45
BODY FLUID pH

Cerebrospinal fluid
Blood plasma
7.35-7.45
7.35-7.45
Cerebrospinal fluid 7.35-7.45
saliva 6-8
saliva
urine 6-8
4-8
Gastric juice 1
urine
Pancreatic juice 4-8
7.5-8

Gastric juice 1
Pancreatic juice 7.5-8
• Enzymatic activity and protein
structure are frequently sensitive
to pH
• In any given body or cellular
compartment, pH is maintained to
allow for maximal enzyme/protein
efficiency
• Solutions having a pH greater
than 7 are alkaline or basic
• The concentration of OH is
-

greater than that of H +

• Conversely, solutions having a

pH less than 7 are acidic
The pH values of some common aqueous fluids

• A cola drink (pH

3.0) has an H +

concentration
approximately
10,000 times
that of blood
(pH 7.4)
• Keep in mind that the pH scale is
logarithmic, not arithmetic

• To say that two solutions differ in

pH by I pH unit means that one
solution has ten times the H+
concentration of the other, but it
does not tell us the absolute
magnitude of the difference
 Logarithm

Number Number Index by which

written in written in 10 has been
usual manner exponential raised (log)
manner

10,000,000 10⁷ 7
10,000 10⁴ 4
10 10¹ 1
1 10⁰ 0
0.1 10⁻¹ -1
0.000,0001 10⁻⁷ -7
 True significance of pH
• pH scale is a logarithmic, not arithmetic
scale
• Each degree decrease or increase in pH
denotes 10 times more or less number
of H⁺ respectively
• Changes which appear small on pH
scale are significant in terms of actual
H⁺ concentration
 Biomedical importance of pH
• Normal pH of blood plasma ranges
from 7.35-7.45
• A pH above 7.45 means alkalosis and
a pH below 7.35 is acidosis, both are
pathological conditions
• All biochemical reactions are enzyme
catalyzed
• The catalytic activity of each enzyme is
maximum at its optimum pH
• Biomolecules contain ionizable groups,
and their ionic state depends upon the
surrounding pH
• Cells and organisms maintain a specific
and constant cytosolic pH, keeping the
biomolecules in their optimum state
• Constancy of pH is maintained by
biological buffers
 Calculating pH
• What will be the pH of 0.1 M HCl?
• Assuming that being a strong acid HCl is
completely dissociated, it’s 0.1 M solution
will contain 0.1 or 10⁻¹ grams H⁺ per litre
• pH= - log [H⁺]
• pH = - log [10⁻¹]
= - [- 1]
=1
• The blood plasma has H⁺ concentration
of 3.95 x 10⁻⁸ M/L. Find out it’s pH?
Measurement of pH
• Measurement of pH is one of the
most important and frequently
used procedures in biochemistry
• The pH affects the structure and
activity of biological
macromolecules
• For example, the catalytic
activity of enzymes is strongly
dependent on pH
• Measurements of the pH of
blood and urine are
commonly used in medical
diagnoses
Measurement of pH
• The two most frequently
used methods to determine
the pH of a solution are by
the use of
• indicators
• pH metery
 Indicators
• The indicators may be used in
liquid form or as indicator papers
commonly called pH papers which
are available commercially

• Indicators cannot, however, be

used in case of colored solutions
• Indicators are weakly
dissociating acids or bases and
are used to find out the acidic
or basic reactions of solutions
• Examples include
–Congo red
–Phenolphthalein
 Congo red
• pH range 3-5
• Yellow at pH 3 or less
• Red when pH is 5 or more
Phenolphthalein
• pH range of 8.3-10
• Colorless at 8.3 or less
• Red at pH 10 or more
• Decreasing or increasing pH beyond this
range will have no further effect
• The electrometric method of
determining pH specially that
involving the use of commercially
available pH meters is very
accurate
• Two electrodes, a glass electrode
and a calomel electrode, are
dipped in unknown solution
• The calomel electrode acts
as the reference electrode

• It contains mercury-
mercurous chloride paste
which is surrounded by KCI
solution
• The glass electrode is a glass bubble
containing known concentration of
HCI (usually 0.1 M)
• When it is dipped into a solution of
unknown pH, a potential difference is
set in proportion to the difference in
the concentration between H+ ions
inside the glass electrode and H+ ions
in the solution with the unknown pH
• The potential difference in the
glass electrode with reference to
the calomel electrode is first
amplified and then measured with
a potentiometer
• The pH value of the unknown
solution is read directly from a
calibrated scale
pHmeter
 RECAP
• Equilibrium constant , concept of Keq
• Ionization and ionic product of H₂O ,
concept of kw
• Hydrogen ion concentration
• Concept of pH
• Significance of pH
• Measurement of pH
• Acid dissociation constant Ka and
pKa
• Henderson Hasselbach equation
• Buffers
• Titration curve of weak acids
Weak Acids and Bases Have
Characteristic Acid Dissociation
Constants
• Hydrochloric, sulfuric, and nitric acids,
commonly called strong acids, are
completely ionized in dilute aqueous
solutions
• The strong bases NaOH and KOH are
also completely ionized
• Of more interest is the behavior of weak
acids and bases-those not completely
ionized when dissolved in water
• These are ubiquitous in biological
systems and play important roles in
metabolism and its regulation
• The behavior of aqueous solutions of
weak acids and bases is best understood
if we first define some terms
• Acids may be defined as proton donors and
bases as proton acceptors
• A proton donor and its corresponding proton
acceptor make up a conjugate acid-base pair
• Acetic acid (CH3COOH), a proton donor, and
the acetate anion (CH3COO-), the
corresponding proton acceptor, constitute a
conjugate acid-base pair, related by the
reversible reaction
CH3COOH  CH3COO- + H+
• Each acid has a characteristic tendency to lose
its proton in an aqueous solution
• The stronger the acid, the greater its tendency
to lose its proton
• The tendency of any acid (HA) to lose a proton
and form its conjugate base (A-) is defined by
the equilibrium constant (Keq) for the
reversible reaction
HA  H+ + A -
Keq = [H+][A-]/[HA] = Ka
• Equilibrium
constants for
ionization
reactions are
usually called
ionization
constants or
acid
dissociation
constants, often
designated Ka
• Stronger acids, such as
phosphoric and carbonic acids,
have larger ionization constants
• Weaker acids, such as
monohydrogen phosphate
(HPO4 ), have smaller ionization
2-

constants
Titration Curves Reveal the ph of
Weak Acids
• Titration is used to determine the
amount of an acid in a given solution
• A measured volume of the acid is
titrated with a solution of a strong
base, usually sodium hydroxide
(NaOH), of known concentration
• The NaOH is added in small
increments until the acid is
consumed (neutralized), as
determined with an indicator dye or
a pH meter
• The concentration of the acid in the
original solution can be calculated
from the volume and concentration
of NaOH added
• A plot of pH against the amount of NaOH
added (a titration curve) reveals the pKa
of the weak acid
• Consider the titration of a 0.1M solution
of acetic acid with 0.1M NaOH at250C
• Two reversible equiIibria are involved in
the process (here, for simplicity, acetic
acid is denoted HAc)
• H2O  H+ + OH
• HA  H+ + A -
• The equilibria must
simultaneously conform to their
characteristic equilibrium
constants, which are,
respectively,
K
• w  [H +
][OH -
] = 1 x 10 -14
M 2

• Ka  [H ][Ac ]/[HAc]
+ -

= 1.74 x 10 M
-5
• At the beginning of the titration, before any
NaOH is added, the acetic acid is already
slightly ionized, to an extent that can be
calculated from its ionization constant
• As NaOH is gradually introduced, the added
OH combines with the free H+ in the solution
to form H2O, to an extent that satisfies the
equilibrium relationship in the equation
• As free H+ is removed, HAc dissociates further
to satisfy its own equilibrium constant
• The net result as the titration proceeds
is that more and more HAc ionizes,
forming Ac-, as the NaOH is added
• At the midpoint of the titration, at
which exactly 0.5 equivalent of NaOH
has been added, one-half of the original
acetic acid has undergone dissociation,
so that the concentration of the proton
donor, [HAc], now equals that of the
proton acceptor, [Ac-]
• At this midpoint a very
important relationship
holds:
• The pH of the equimolar
solution of acetic acid and
acetate is exactly equal to
the pKa of acetic acid (pKa
= 4.76)

• As the titration is
continued by adding
further increments of
NaOH, the remaining
nondissociated acetic acid
is gradually converted into
• The end point of the titration occurs
at about pH 7.0:
• All the acetic acid has lost its
protons to OH-, to form H2O and
acetate
• Throughout the titration the two
equilibria coexist, each always
conforming to its equilibrium
constant
• Figure compares
the titration
curves of three
weak acids with
very different
ionization
constants
• acetic acid (pKa =
4.76); dihydrogen
phosphate, H2PO,
(pKa = 6.86); and
ammonium ion,
NH4, (pKa = 9.25)
• Although the titration curves of these
acids have the same shape, they are
displaced along the pH axis because the
three acids have different strengths

• Acetic acid, with the highest Ka (lowest

pKa) of the three, is the strongest of the
three weak acids (loses its proton most
readily); it is already half dissociated at
pH 4.76
• Dihydrogen phosphate loses a proton less
readily, being half dissociated at pH 6.86
• Ammonium ion is the weakest acid of the
three and does not become half
dissociated until pH 9.25
• The titration curve of a weak acid shows
graphically that a weak acid and its anion-
a conjugate acid-base pair-can act as a
buffer
Buffering against pH Changes In Biological Systems
• Almost every biological process is pH-dependent; a
small change in pH produces a large change in the
rate of the process
• This is true not only for the many reactions in which
the H+ ion is a direct participant, but also for those
in which there is no apparent role for H+ ions
• The enzymes that catalyze cellular reactions, and
many of the molecules on which they act, contain
ionizable groups with characteristic pKa values
• The protonated amino and carboxyl groups of
amino acids and the phosphate groups of
nucleotides, for example, function as weak acids;
their ionic state is determined by the pH of the
surrounding medium
• When an ionizable group is sequestered in the
middle of a protein, away from the aqueous
solvent, its pKa, or apparent pKa, can be
significantly different from its pKa in water
• As we noted above, ionic interactions are among
the forces that stabilize a protein molecule and
allow an enzyme to recognize and bind its substrate
• Cells and organisms maintain a specific
and constant cytosolic pH, usually near
pH 7, keeping biomolecules in their
optimal ionic state
• In multicellular organisms, the pH of
extracellular fluids is also tightly
regulated
• Constancy of pH is achieved primarily by
biological buffers: mixtures of weak acids
and their conjugate bases
The Henderson-Hassebalch Equation Relates
pH, pka, and Buffer Concentration
• This equation is simply a useful way of
restating the expression for the ionization
constant of an acid
• For the ionization of a weak acid HA, the
Henderson-Hasselbalch equation can be
derived as follows:
Ka = [H+][A-]/[HA]
• First solve for [H+]:
[H+] = ka[HA]/[A-]
• Then take the negative logarithm of both
sides:
-log[H+] = -logka - log[HA]/[A-]
• Substitute pH for -log [H+] and pKa for -log Ka
pH = pka – log[HA]/[A-]
• Now invert -log [HA]/[A-], which involves
changing its sign, to obtain the Henderson-
Hasselbalch equation:
pH = pka + log[A-]/[HA]
• This equation enables us to deduce some
important quantitative relationships
• For example, it shows why the pKa of a
weak acid is equal to the pH of the
solution at the midpoint of its titration
• At that point,
[HA] = [A-]
pH = pka + log 1 = pKa + 0
• The Henderson-Hasselbalch equation
also allows us to
(1) calculate pKa, given pH and the molar
ratio of proton donor and acceptor
( 2) calculate pH, given pK, and the molar
ratio of proton donor and acceptor
(3) calculate the molar ratio of proton
donor and acceptor, given pH and pKa
 Solving problems using HH equation
• Calculate the pKa of lactic acid, given that when the
concentration of lactic acid is 0.01M and the
concentration of lactate is 0.087 M, the pH is 4.80
• pH= pKa + log [lactate]/[lactic acid]
• pka= pH- log [lactate]/[lactic acid]
= 4.80 - log 0.087/0.010
= 4.80 - log 8.7
= 4.80 - 0.94
= 3.9 (answer)
• Calculate the ratio of the concentrations
of acetate and acetic acid required in a
buffer system of pH 5.30 (pKa = 4.76)
• pH= pKa + log [acetate]/[acetic acid]
• log [acetate]/[acetic acid]= pH- pKa
= 5.30-4.76
= 0.54
• [acetate]/acetic acid] = antilog 0.54
= 3.5
• An important buffer system of
plasma is NaHCO₃/H₂CO₃. Their ratio
under normal circumstances is 20/1.
Find out the pH of plasma. The pKa
of H₂CO₃ is 6.1.
• pH = pKa + log [salt]/[acid]
• = 6.1 + log 20/1
• = 6.1+ 1.3 = 7.4
• Three primary systems regulate the
H concentration in the body fluids
+

to prevent acidosis or alkalosis

1. The chemical acid-base buffer

systems of the body fluids, which
immediately combine with acid or
base to prevent excessive changes
in H+ concentration
2. The respiratory center, which
regulates the removal of CO2 (and,
therefore, H2CO3) from the
extracellular fluid

3. The kidneys, which can excrete either

acid or alkaline urine, thereby
readjusting the extracellular fluid H+
ion concentration toward normal
during acidosis and alkalosis
 The buffer systems
• When there is a change in H+ ion
concentration, the buffer systems of
the body fluids react within seconds to
minimize these changes
• Buffer systems do not eliminate
hydrogen form or add them to the body
but only keep them tied up until
balance can be reestablished
• The second line of defense, the
respiratory system acts within a few
minutes to eliminate CO2, and
therefore H2CO3 from the body
• These first two lines of defense keep
the H+ ion concentration from
changing too much until the more
slowly responding third line of
defense, the kidneys, can eliminate
the excess acid or base from the body
• Although the kidneys are
relatively slow to respond
compared with other
defenses, over a period of
hours to several days, they
are by far the most powerful
of the acid base regulatory
systems
 Buffers Are Mixture of Weak Acids
and Their Conjugate Bases
• Buffers are aqueous systems that
tend to resist changes in pH when
small amounts of acid (H+) or base
(OH ) are added
-

• A buffer system consists of a weak

acid (the proton donor) and its
conjugate base (the proton acceptor)
• As an example, a
mixture of equal
concentrations of acetic
acid and acetate ion,
found at the midpoint
of the titration curve, is
a buffer system
• Notice that the titration
curve of acetic acid has
a relatively flat zone
extending about 1 pH
unit on either side of its
midpoint pH of 4.76
• In this zone, a given amount of H+
or OH added to the system has
-

much less effect on pH than the

same amount added outside the
zone
• This relatively flat zone is the
buffering region of the acetic acid-
acetate buffer pair
• At the midpoint of the buffering
region, where the concentration of
the proton donor (acetic acid) exactly
equals that of the proton acceptor
(acetate), the buffering power of the
system is maximal; that is, its pH
changes least on addition of H or OH
+ -

• The pH at this point in the titration

curve of acetic acid is equal to its pKa
• The pH of the acetate buffer system does
change slightly when a small amount of
H+ or OH- is added, but this change is very
small compared with the pH change that
would result if the same amount of H+ or
OH- were added to pure water or to a
solution of the salt of a strong acid and
strong base, such as NaCl, which has no
buffering power
• Each conjugate acid-base pair has a
characteristic pH zone in which it is an
•
effective buffer
• The H2PO4-/HPO42- pair has a pKa of 6.86
and thus can serve as an effective buffer
system between approximately pH 5.9
and pH 7.9
• The NH 4 /NH3 pair, with a pKa of 9.25,c
+

an act as a buffer between approximately

pH 8.3 and pH 10.3
 Buffering capacity
• The capacity of a buffer to absorb
H⁺ or OH⁻ions depends mainly
upon the concentration of the
buffering components
• It also depends upon the ratio of
salt to acid concenteration
• Efficiency of a buffer also
depends upon the pH
The buffer value or capacity
• The smaller the pH change
caused by the addition of a given
amount of acid or alkali, the
greater is the buffer capacity
 Weak Acids or Bases Buffer Cells and
Tissues against pH Changes
• The intracellular and extracellular
fluids of multicellular organisms have
a characteristic and nearly constant
pH
• The organism's first line of defense
against changes in internal pH is
provided by buffer systems
 Buffering in the Blood
• The major source of acids in the
blood under normal conditions is
through cellular metabolism

• Amino acids are utilized in the liver

for gluconeogenesis, leaving NH4+
and HCO3– as products from their
amino and carboxyl groups
• The NH 4 is incorporated into urea and
+

the protons that are formed are buffered

intracellularly by HCO3–, so little NH4+ and
HCO3– escape into the circulation

• However, metabolism of sulfur-containing

amino acids produces H2SO4, and
metabolism of phosphorylated amino
acids such as phosphoserine produces
H3PO4
• These strong acids enter the circulation
and present a major H+ load to the
buffers in the ECF
• The H+ load from amino acid
metabolism is normally about 50 mEq/d
• The CO2 formed by metabolism in the
tissues is in large part hydrated to
H2CO3, and the total H+ load from this
source is over 12,500 mEq/d
• However, most of the CO2 is excreted in
the lungs, and only small quantities of the
H+ remain to be excreted by the kidneys
• Common sources of extra acid loads are
– Strenuous exercise (lactic acid)
– Diabetic ketosis (acetoacetic acid and -
hydroxybutyric acid)
– Ingestion of acidifying salts such as NH4Cl
and CaCl2,
• Fruits are the main dietary source
of alkali

• They contain Na+ and K+ salts of

weak organic acids, and the anions
of these salts are metabolized to
CO2, leaving NaHCO3 and KHCO3 in
the body
• Such ingestion contributes little to
changes in pH and a more
common cause of alkalosis is loss
of acid from the body as a result
of vomiting of gastric juice rich in
Hill
• This is, of course, equivalent to
adding alkali to the body
• Acid and base shifts in the blood
are largely controlled by three
main buffers in blood:
(1) proteins
(2) hemoglobin
(3) the carbonic acid–
bicarbonate system
• Plasma proteins are effective
buffers because both their free
carboxyl and their free amino
groups dissociate:
RCOOH  RCOO- + H+
RNH3  RNH2 + H
+ +
• The second buffer system is
provided by the dissociation of
the imidazole groups of the
histidine residues in hemoglobin
• In the pH 7.0–7.7 range, the free
carboxyl and amino groups of
hemoglobin contribute relatively
little to its buffering capacity
• However, the hemoglobin molecule contains
38 histidine residues, and on this basis—plus
the fact that hemoglobin is present in large
amounts—the hemoglobin in blood has six
times the buffering capacity of the plasma
proteins
• The action of hemoglobin is unique because
the imidazole groups of deoxyhemoglobin
(Hb) dissociate less than those of
oxyhemoglobin (HbO2), making Hb a weaker
acid and therefore a better buffer than HbO2
• The third and major buffer system in
blood is the carbonic acid–bicarbonate
system:
• H2CO3  H+ + HCO3-

• The Henderson–Hasselbalch equation

for this system is
• pH  pK + log [HCO3-]/[H2CO3]
• The pK of this system is low relative
to the pH of the blood, but the
system is one of the most effective
buffer systems in the body because
the amount of dissolved CO2 is
controlled by respiration
• Additional control of the plasma
concentration of HCO3– is provided
by the kidneys
• When H+ is added to the blood,
HCO3 declines as more H2CO3 is
–

formed

• If the extra H2CO3 were not

converted to CO2 and H2O and the
CO2 excreted in the lungs, the
H2CO3 concentration would rise
• When enough H+ has been added to
halve the plasma HCO3–, the pH would
have dropped from 7.4 to 6.0
• However, not only is all the extra H2CO3
that is formed removed, but also the H+
rise stimulates respiration and therefore
produces a drop in PCO2, so that some
additional H2CO3 is removed
• The pH thus falls only to 7.2 or 7.3
• There are two additional factors
that make the carbonic-acid-
bicarbonate system such a good
biological buffer
• First, the reaction CO2 + H2O ⇄
H2CO3 proceeds slowly in either
direction unless the enzyme
carbonic anhydrase is present
• There is no carbonic anhydrase in
plasma, but there is an abundant
supply in red blood cells
• Second, the presence of hemoglobin
in the blood increases the buffering
of the system by binding free H+
produced by the hydration of CO2
and allowing for movement of the
HCO3 into the plasma
–
• Buffering in vivo is, of course, not limited to the
blood
• The principal buffers in the blood, interstitial fluid,
and intracellular fluid are listed below
Blood:
1. Bicarbonate
2. Proteins
3. Haemoglobin
Interstitial fluid:
4. Bicarbonate buffer
Intracellular fluid:
1. Proteins
2. Phosphate
Cerebrospinal fluid (CSF):
3. Bicarbonate
4. Phosphate systems
Renal and Urinary buffers:
5. Bicarbonate
6. Phosphate
7. Ammonia
 Bicarbonate buffer
NaHCO₃ /H₂CO₃
• H₂CO₃ is formed by the following reaction
CO₂ + H₂O ↔ H₂CO₃
• H₂CO₃ ionizes weakly:
H₂CO₃↔ H⁺+ HCO₃⁻
• NaHCO₃ ionizes completely:
NaHCO₃ → Na ⁺+ HCO₃⁻
• Putting the entire system together:
CO₂ + H₂O ↔ H₂CO₃ ↔ H⁺ + HCO₃⁻
• When a strong acid is added to the system:
↑H⁺+ HCO₃⁻ → H₂CO₃→ CO₂ + H₂O
• When a strong base is added to the system:
NaOH + H₂CO₃ → NaHCO₃ + H₂O
 Quantitative dynamics of the bicarbonate
buffer
• H₂CO₃ ↔ H⁺+HCO₃⁻

• pKa for the system is 6.1

• Buffering zone is 5.1-7.1
• Molar concentration of
HCO₃-/ H₂CO₃= 20/1
• Plasma concentration is low, but powerful
buffer
 Phosphate buffer system
• Plays a major role in buffering renal tubular
fluid and intracellular fluids
• Main elements are H₂PO₄⁻ and HPO₄⁻²
• When a strong acid is added to phosphate
buffer solution:
HCl + Na₂HPO₄ → NaH₂PO₄ + NaCl
• The strong acid , HCl is replaced by a weak
acid NaH₂PO₄
• When a strong base NaOH is added to the
system
NaOH + NaH₂PO₄ → Na₂HPO₄ + H₂O
• pKa is 6.8, which is close to 7.4 , allowing
this buffer to operate near it’s maximum
buffering power
• However it is a weak buffer, as its
concentration in the extra cellular fluid is
low (8% the concentration of bicarbonate
buffer)
• However, it is important in important in
the tubular fluid of kidneys because:
1. Phosphate usually becomes greatly
concentrated in the tubules, thereby
increasing the buffering power of the
phosphate system
2. The tubular fluid has a lower pH than
E/C fluid, bringing the operating range
of buffer, pKa (6.8) closer to the
surrounding pH
 Protein buffer system
• About 60-70% of the total chemical
buffering of the body fluids is inside the
cells, and most of it results from the
intracellular proteins, because:
1. High concentration of proteins
intracellularly
2. Pka of protein systems close to 7.4
(Histidine has a pKa of 6)
Acidosis & Alkalosis
• The pH of the arterial plasma is
normally 7.40 and that of venous
plasma slightly lower
• Acidosis:A decrease in pH below
the norm is technically present
whenever the arterial pH is
below 7.40
• Alkalosis: An increase in pH is
technically present whenever
pH is above 7.40
• In practice, variations of up to
0.05 pH unit occur without
untoward effects
• Acid–base disorders are split into four
categories:
1. respiratory acidosis
2. respiratory alkalosis
3. metabolic acidosis
4. metabolic alkalosis

• In addition, these disorders can occur in

combination
Use of Anion Gap to Diagnose Acid-Base
Disorders
• The concentrations of anions and cations in
plasma must be equal to maintain electrical
neutrality
• Therefore, there is no real “anion gap” in the
plasma
• However, only certain cations and anions are
routinely measured in the clinical laboratory
• The cation normally measured is Na+, and the
anions are usually Cl– and HCO3–
• The “anion gap” (which is only a
diagnostic concept) is the difference
between unmeasured anions and
unmeasured cations, and is
estimated as

• Plasma anion gap

= [Na+] – [HCO3–] – [Cl–]
= 144 – 24 – 108 = 10 mEq/L
• The anion gap will increase if unmeasured
anions rise or if unmeasured cations fall
• The most important unmeasured cations
include calcium, magnesium, and potassium,
and the major unmeasured anions are
albumin, phosphate, sulfate, and other
organic anions
• Usually the unmeasured anions exceed the
unmeasured cations, and the anion gap
ranges between 8 and 16 mEq/L
• The plasma anion gap is used mainly in
diagnosing different causes of metabolic
acidosis
• In metabolic acidosis, the plasma HCO3–
is reduced
• If the plasma sodium concentration is
unchanged, the concentration of anions
(either Cl– or an unmeasured anion) must
increase to maintain electroneutrality
• If plasma Cl– increases in proportion to
the fall in plasma HCO3–, the anion gap
will remain normal, and this is often
referred to as hyperchloremic metabolic
acidosis
• If the decrease in plasma HCO3– is not
accompanied by increased Cl–, there must
be increased levels of unmeasured anions
and therefore an increase in the
calculated anion gap
• Metabolic acidosis caused by excess
nonvolatile acids (besides HCl), such as
lactic acid or ketoacids, is associated
with an increased plasma anion gap
because the fall in HCO3– is not matched
by an equal increase in Cl–
• By calculating the anion gap, one can
narrow some of the potential causes of
metabolic acidosis
THE COLLOIDAL STATE
• The word colloid was first used for solutions of
certain substances such as proteins, starch and
gums which do not diffuse through most of the
membranes
• Substances which diffuse readily through
membranes were called crystalloids
• Now it is known that this property is only
determined by the size of the solute particles
• Colloidal particles may be single ions, molecules or
aggregates of many molecules
 True solution particles in a solution
• When dissolved, they
disintegrate into
individual ions or
molecules which are less
then 1 nm in diameter
• They are not visible even
with electron
microscope, with which
particles as small as 1
nm can be seen
 Colloidal particles
• In this case the size of
the particles ranges
from 1 to 100 nm or
according to some
authors even to 500 nm
• They can be seen with
the help of electron
microscope and can
also be made visible by
the ultramicroscope
Colligative Properties of
Solutions
• The term colligative means
tied together and colligative
properties are those which are
simultaneously bestowed
upon a solvent by its content
of solute particles
• The following four changes in the
colligative properties are seen as the
number of particles increase in a
solution
1. The osmotic pressure is increased
2. Boiling point is raised
3. Freezing point is depressed
4. Vapor pressure is decreased
• As the particles in a colloidal solution are
much bigger than those in a true solution,
a colloidal solution will contain only a
minute fraction of the number of particles
present in a true solution of the same
strength
• For this reason, the above mentioned
changes are minimal with a colloidal
solution because these properties depend
only on the number of particles in the
solution
 Properties of Colloidal Solutions
1. Sols and gels
• A colloidal solution
when it exists in
liquid form is called a
sol
• Many lyophilic
colloids can be
changed into a
semisolid form which
is called a gel
2. Thixotropy
• The structure of a gel may
be broken by shaking or
stirring and on standing a
gel may be formed again
• This phenomenon of
reversible gel - sol
transformation is termed
thixotropy
• It is thought that
thixotropy occurs in
colloidal systems having
non-spherical particles
3. Imbibition
• It is a process by which
dry proteins take up
water and retain it
• The water can be
squeezed out only with
great difficulty and
pressure
• Seeds take up water with
great ease as their
proteins are in a
dehydrated state
• Imbibition is greatly increased in the
presence of Na+, K+ and Ca+ ions
• Plasma albumin can hold 17 ml
water/gram and is the chief agent in
keeping the plasma volume constant by
not letting plasma water to leave the
lumen of blood capillaries
• For this reason, it is very useful as a
plasma expander and is sometimes
transfused as a substitute for blood
plasma
4. Brownian movement
• If we see a colloidal solution constantly through
an ultramicroscope, the particles are seen to be in
a zig-zig motion
• This is called Brownian movement, and is due to
an unequal and random bombardment of the
particles by the molecules of the solvent
• The Brownian movement is more, if the particle is
of small size and the solvent is less viscous
• A rise in temperature also increases Brownian
movement
5. Tyndall effect
• In the ultramicroscope an intense beam of
light is passed transversely through the
colloidal solution, which is then viewed at
right angle to the path of incident light
• The colloidal particles appear as bright spots
against a dark background and the path of the
beam of light passing through a colloidal
solution becomes plainly visible as milky
turbidity
• This
phenomenon is
known as
Tyndall effect
and is due to the
reflection and
scattering of the
light by the
particles present
in the solution
Adsorption
• Adsorption is the
phenomenon by
which a layer of ions,
molecules or
aggregates of
molecules that are
present in a medium
is condensed upon a
surface with which
they come in contact
• Activated charcoal (charcoal heated in
steam and then to 7000 to 8000C), silica
gel, kaolin, alumina cream and Fuller’s
earth are the commonly used adsorbents
• Adsorption is a reversible phenomenon,
it decreases with rise in temperature and
increases with rise in pressure
• The mechanism of adsorption is still
unclear
• The adsorbing property of an adsorbent is
much increased by dividing it into small-
sized particles that serve to increase the
surface area relative to the volume
• Thus more area becomes available for
adsorption to take place
• Activated charcoal is one of the best
adsorbents, because one gram of it has been
found to have a surface area around 120
square meters
• Adsorption is a very important
phenomenon and finds many
applications
• Adsorbents such as charcoal are
used to decolorize many solutions
• Charcoal is also used in gas masks
because it takes up obnoxious
gases
• It is also used for the adsorption of
excessive gas in the gastrointestinal
tract and for the treatment of
poisoning
• Drugs like kaolin used in diarrhea owe
their actions to adsorption of
bacterial toxins
• Adsorbents are also used in the
purification of many proteins
SURFACE TENSION
• The surface of a liquid behaves as if it
were a stretched elastic membrane which
resists external forces without rupture
and exerts a pressure inwards on the rest
of the liquid
• This force causes a liquid to assume the
form of a spherical drop in which form it
has the smallest surface area for a given
volume
• The molecules of a
solution are strongly
attracted to each
other, but only to a
very short distance
• Surface tension is the
force or tension
required to break this
film and is defined as
the force in dynes
acting upon a line one
cm long on the surface
• Surface tension is measured as
ergs/cm2 or as dynes/cm

• Soaps, detergents, bile salts

and proteins reduce the
surface tension, while salts like
NaCl increase it
• Bile salts, on account
of their surface
tension lowering
property, greatly
facilitate the
emulsification and
digestion of fats in the
small intestine
• The surface tension is
also usually decreased
by an increase in
temperature
Donnan Effect
• When an ion on one side of a membrane
cannot diffuse through the membrane,
the distribution of other ions to which
the membrane is permeable is affected in
a predictable way
• For example, the negative charge of a
nondiffusible anion hinders diffusion of
the diffusible cations and favors diffusion
of the diffusible anions
• Consider a situation, in which a membrane
(m) between compartments X and Y is
impermeable to charged proteins (Prot–) but
freely permeable to K+ and Cl–
• Assume that the concentrations of the anions
and of the cations on the two sides are
initially equal
• Cl– diffuses down its concentration gradient
from Y to X, and some K+ moves with the
negatively charged Cl– because of its opposite
charge
• Donnan and Gibbs showed that in the
presence of a nondiffusible ion, the
diffusible ions distribute themselves
so that at equilibrium their
concentration ratios are equal
• This is the Gibbs–Donnan equation
• It holds for any pair of cations and
anions of the same valence
• The Donnan effect on the distribution of ions
has three effects in the body
• First, because of charged proteins (Prot–) in
cells, there are more osmotically active
particles in cells than in interstitial fluid, and
because animal cells have flexible walls,
osmosis would make them swell and
eventually rupture if it were not for Na, K
ATPase pumping ions back out of cells
• Thus, normal cell volume and pressure
depend on Na, K ATPase
• Second, because at equilibrium the distribution of
permeant ions across the membrane is
asymmetric, an electrical difference exists across
the membrane
• The charges line up along the membrane, with the
concentration gradient for Cl– exactly balanced by
the oppositely directed electrical gradient, and
the same holds true for K+
• Third, because there are more proteins in plasma
than in interstitial fluid, there is a Donnan effect
on ion movement across the capillary wall
VISCOSITY
• Flowing is one of the
characteristic properties of
liquids and this property is
termed fluidity
• Some liquids flow more readily
than other because they have
more fluidity
• The reciprocal of fluidity is called
viscosity which means the resistance
offered by a liquid to flow; this
resistance to flow depends on the
friction of its component molecules as
the flow past one another
• Viscosity is believed to result from the
attraction of the molecules between
one another as well as from asymmetry
in their structure
• Thus water will flow over a glass plate
much fast than honey and therefore
water is said to have more fluidity but
less viscosity than honey
• The rate of flow depends on the nature of
the liquid and on the force which
produces the flow
• The liquid may be thought to consist of a
number of molecular layers arranged one
over the other
• The displacement of different layers
relative to one another is opposed by
the internal friction or viscosity of the
liquid

• The rate of flow of a liquid is inversely

proportional to its viscosity; in other
words, higher the viscosity, lesser will
be the flow
 Factors affecting viscosity
1. Temperature: viscosity decreases 2% with each
degree centigrade rise in temperature
2. Chemical composition: Viscosity is greater with
larger and elongated molecules
3. Presence of colloidal particles: the viscosity of
lyophilic colloids is relatively high
4. Effect of suspended material: suspended
materials increase viscosity in proportion to the
volume of the suspended material
 Clinical application
• The viscosity of blood is 4.5 times that of
water
• It is important in blood flow and pressure
• The relative viscosity of serum is 1.4-1.8
times that of water
• When it goes between 4 and 7 owing to
the presence of abnormal proteins
hyperviscosity syndrome results
• The viscosity of a liquid
is determined by
comparison with water
• One method of
determining the
viscosity of a liquid is by
the use of Ostwald’s
viscosimeter or
viscometer, which
consists of a U-tube
bearing two bulbs in one
arm and a capillary tube
of a suitable bore
 Measurement of viscosity
• Viscosity is determined in comparison to
water.

• Ostwald’s vicosimeter.
• Relative viscosity of a liquid =
• density of liquid x T₂
density of water x T₁
T₁ = time in seconds taken by water from
upper to lower mark.
T₂ = time in seconds taken by a liquid
from upper mark to lower mark.
0.8 x 120 = 1.6
1 x 60
• Unit of viscosity : poise
• expressed in dyne sec per cm².

• The absolute viscosity of water at 25°C

is 0.00895 poise.
Chloride shift