Concept of pH and

buffer
Dr. Zakiah Jubri

Objectives
At the end of the lecture, students
should be able to:

describe the concept of ionization, pH and

pKa
define acids, bases and their relation in
normal metabolism.
explain the Henderson-Hasselbalch
equation.
explain the principle of buffer and its
function in human body

References

Principles of Biochemistry. Horton HR., Moran LA.,

Ochs RS., Rawn JD., Scrimgeour KG. 2002.Third
Edition. Prentice-Hall, Inc, NJ.
Medical Biochemistry. Bhagavan NV. Fourth Edition.
2002. Harcourt Academic Press, San Diego.
Harpers Illustrated Biochemistry. Murray RK.,
Granner DK., Mayes PA., Rodwell VW. Twenty sixth
edition. 2003. Lange Medical Books/McCraw-Hill, NY.
Basic Medical Biochemistry. A clinical approach.
Marks DB., Marks AD., Smith CM. 1996. Lippincott
Williams & Wilkins.

Water
H2O

H+ + OH-

Pure water consists of H2O.

Water molecule can dissociate and ionize to

form hydrogen ions, H+ and hydroxyl ions, OH-.

These 2 products influences cells constituents
H+ determines pH of the medium
Influences catalytic activities of various cellular
enzymes

Dipolar Nature of Water

Two hydrogen atom

linked covalently to an
oxygen atom
Oxygen has a strong
electron-withdrawing
tendency

Lie more towards

oxygen
two hydrogen atoms
partial positive charges.

Oxygen has two more

pairs of electron which

remain unshared
localized partial
negative charge

 Form hydrogen

bonds between
positive pole and
negative pole of
water molecule.

Solvent Properties
Because of its dipolar character

In the crystal structure, NaCl is held together by strong

electrostatic attractions between Na+ and ClIn water, negative pole of the water is attracted towards Na +
and positive pole towards Cl-

 In the human body

contains inorganic cations and anions.

Cations are attracted to the oxygen

atom and anions

hydrogen atoms.

Ion-dipole interactions
hold the components of soluble salts in
solutions.

pH
Measure the proton H+ concentration in moles per liter.
Defines as negative logarithm of hydrogen ion
concentration, expressed in moles
pH = - log H+

The normal pH of human blood -physiological pH is 7.367.44

Diabetes, can have a lower pH, a condition called
acidosis.
Alkalosis
prolonged vomiting (loss of HCl from the stomach)
hyperventilation (excessive loss of carbonic acid as
carbon dioxide).

 Acids - referred as proton donors

Bases -proton acceptors
Strong acids and strong bases

dissociate completely in water, such as HCl and

NaOH.

Weak acids and weak bases.

do not dissociate completely in water.

such as the amino acids (proteins)
purines and pyrimidines (DNA and RNA)
Important in living systems

Relationship between acids

and bases

HCl + H2O

Cl- + H3O+

acid base

base acid

H+ donor

H+ acceptor

Weak acids
CH3COOH
Acetic acid
(weak acid)

conjugate conjugate
base of HCl acid of H2O

H+ + CH3COOAcetate anion
(conjugate base)

Dissociation constant (Ka)

For a weak acid indicates the tendency of the acid
to lose its proton and form its conjugate base
HA
H + + AKa = H+A-
HA

For acetic acid.

Ka = H+CH3COO-
CH3COOH
Ka = 1.76 X 10-5 M at 25oC

Henderson-Hasselbalch
Equation
pH = pKa + log A-
HA
pH = pKa + log Proton acceptor
Proton donor
pH = pKa + log conjugate base
weak acid
pKa value is a measure of the acid strength of a
particular compound. pKa=2 > 5
pH=pKa, a weak acid is 50% dissociates

BUFFERS
Is a system that resists any changes in its pH

when a small amount of acid or alkali is added.

Consists of a weak acid and its conjugate base.
Effectiveness of a buffer
These two components are present in equimolar
concentrations
pH (of the medium) equals to pKa (of the acid-base
pair)

It remains effective between 1 pH unit below

and 1 unit above the pKa

pH 5.76 (Zone
of buffering
capacity)
pH 3.76

 Body fluids must be protected against

changes in pH because most enzymes are

very pH sensitive.
Constantly active because metabolism
involves the production of acids and bases.
Long term, excess acid or base is
eliminated via the kidney and the lungs
Short term, by buffering system

Major buffer systems

pH of the blood is maintained
7.36-7.44

Bicarbonate-carbonic acid buffer system

operates in extracellular fluid

Hemoglobin buffer system

in red blood cells.

Phosphate buffer system

in all types of cells

Protein buffer system

cells and plasma.

Bicarbonate Buffer System

Carbonic acid and

bicarbonate
Is regulated by
kidney and lungs.
CO2

major source of
metabolic acid in
the body
produced from fuel
oxidation in the
TCA cycle

 CO2(d) in blood= CO2(g) in lung

H2CO3 dependent on the PaCO2(g)
H+ increases, it will be taken up by HCO-3

H2CO3 increases CO2(d) in the blood

increases increase in the pressure of CO2 in
the lungs and exhaled through increased rate
of breathing.
H+ reduces, rate of breathing reduces, CO2
retained, form CO2(d) , form H2CO3 , back to
normal.

Bicarbonate and
Hemoglobin

Buffering the blood and transporting CO2

to the lung.
CO2 diffuses interstitial fluid blood
plasma RBC (carbonic anhydrase)

 H+ combine with bicarbonate.

Carbonic acid dissociate into CO2 and
water.
In lung, CO2 releases

Hb loses H+ allows it to bind oxygen

Bicarbonate buffer system is intimately
linked to the delivery of oxygen to
tissues.

 pH falls, breathe more rapidly and expire more

CO2.
pH rises, breathe more shallowly. CO 2 retained
form carbonic acid back to normal pH
Rate of breathing contributes in pH regulation
dissolved CO2 in blood.
Bicarbonate and carbonic acid buffer system for
plasma and interstitial fluid.
Blood extracellular proteins, buffering capacity
through amino acid side chains accept and
release protons

Phosphate anions and

proteins
Maintaining a constant
pH of intracellular
fluids.
Phosphate anions

Int buffer in RBC and cells

Conc is much higher than
in blood and interstitial
fluid

Organic phosphate
anions

Glucose 6-phosphate and

ATP

Protein- histidine and

amino acids
Accept H+

 Acetoacetic acid and -hydroxybutyric acid

From fatty acid oxidation to ketone bodies

Lactic acid
Glycolysis

Cell becomes too acidic,

more H+ is transported out in exchange for Na+ ions

Cell becomes too alkaline,

more bicarbonate is transported out in exchange for Clions.

THANK YOU