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Unit-II : Water, PH, Buffer

 Introduction - Properties of water - Role of water in biological system Dr. A.D.NAVEEN KUMAR
 Acids and Bases – pH – pKa – pKb - Acid-base balance CMHS

 Water is an important solvent in which chemical reactions of biological processes
A.D.NAVEEN evolved.
 All aspects of cell structure and function are adapted to the physical and chemical
CHMSKUMAR, CHMSproperties of
 Water accounts for 70% of the weight of most organisms.
 The strong attractive forces between water molecules result in waters solvent properties.
 The H2O molecules and its ionization products – H+ and OH- ions greatly influence the structure.
and functions of all cellular compartments such as Enzymes, Proteins, Nucleic acids and Lipids.
Water is needed not only for Biochemical reactions of cell but also for:
 Transporting substances across the membranes.
 Maintenance of intracellular pH.
 Maintenance of osmolarity with electrolytes.
 Maintaining body temperature (370C).
 Dissolving and excretion of waste products.
 Producing digestive fluids.

H2O is the most abundant substance on earth and can exists in three states – Liquid, Solid and Gaseous.

In H2O the hydrogen atoms (H) have a partial positive charge and the oxygen atoms (O) have partial
negative charge.

Water is a dipole because of its geometry and the difference in electro negativity between hydrogen and
oxygen. Oxygen is more electronegative than hydrogen.

The polar nature and geometry of the water molecule allows water molecules to form hydrogen
bonds with each other and with dissolved hydrophilic substances.

Hydrogen bonds between water molecules = electrostatic attraction between the oxygen atom of
one water and the hydrogen of another.

Water can also form hydrogen bonds with functional groups of hydrophilic (polar or ionic)
biomolecules and organic compounds.

Each H2O molecule can form four Hydrogen bonds to other molecules, resulting in a tetrahedral
Distribution of Water in Human body:

Acids, Bases and Buffers :

Acids- releases proton in water (proton donor).
HA H+ + A-
Acid Proton Conjugate base
Base- accepts proton in water (proton acceptor) or releases hydroxyl ions (OH-) in water.
BOH OH- + B+
Base Proton Conjugate acid
Each acid has a characteristic tendency to lose its proton(H+) in an aqueous solution. The stronger the
acid, the greater its tendency to lose its proton (H+) but the weak acids release their protons slowly.

Examples of Strong acids: HCl, H2SO4, HNO3 (Inorganic acids) - Dissociate completely in water.
Examples of Weak acids: Acetic acid, Lactic acid, Phosphoric acid, Carbonic acid and Citirc acid
(Organic acids). Weak acids are common in biological systems and produced during metabolic reactions.

Examples of Strong bases: NaOH, KOH, Ba(OH2)

Examples of Weak bases: Amines, NH4+, Aniline, Pyridine

The tendency of any acid (HA) to lose proton and form its conjugate base is defined as Dissociation
constant (Ka). The dissociation constant of acid can be written as

pKa = log 1 / Ka = -log Ka

pKa is a quantitative measure of acid strength.

 Smaller pKa (<1) refers to Stronger acid – the stronger tendency to dissociate a Proton.
 Larger pKa (>1) refers to Weaker acid - the weaker tendency to dissociate a Proton.

pKb refers to the strength of Bases.

pH :
 Sorenson introduced the pH scale in 1909 using the symbol pH ranging from 0 – 14.
 pH refers to the H+ ion concentrations.
 The concentration of H+ must be expressed in Molar terms i.e. Moles/Litre.

The term pH is defined by the expression of pH = log 1 / [H] = -log [H+]
 p - denotes the negative logarithm of H+.
 In pH scale 0 - 7 refers to Acidic, 7 refers to Neutral and 7 – 14 refers to Basic or Alkali.
For a precisely neutral solution at 250C, in which the concentration of Hydrogen ions is 1.0 x 10-7M, the
pH can be calculated as follows pH = log 1 / [1.0 x 10-7] = 7.0

pH values of Human body fluids and secretions :

Body fluid/secretion pH
HCl of Gastric juice 0.87
(parietal secretion)
Urine 6.0
Milk 6.6 - 6.8
Saliva 7.2
Aqueous humor of eye 7.2
Blood 7.4
Cerebrospinal fluid 7.4
Intestinal juice 7.6 - 7.8
Bile 7.6 - 7.8
Pancreatic Juice 8.0

Buffers :
Buffers are aqueous systems that tend to resist or prevent the changes in pH of the solution upon addition
of acid(H+) or base (OH-) ions are added.

Buffers are extremely important in biological systems.

Examples: Maintaining blood pH; maintaining physiological pH inside cells

Buffers are Mixtures of Weak acids (Proton donor) and their Conjugate base (Proton acceptor).

Example: Acetic acid (CH3COOH) and Acetate ion (CH3COO-)

Henderson-Hasselbalch Equation:
The quantitative relationship among pH, buffering action of a mixture of weak acid with its conjugate
base, and the pKa of the weak acid is given by a simple expression called Henderson-Hasselbalch

It allows us to calculate pKa, given pH and the molar ratio of proton donor and acceptor.
In human body two major types of Buffer systems are responsible for the maintenance of pH.

1). Bicarbonate Buffer System :

It is the major buffer system responsible for Extracellular fluid (Blood) pH maintenance-7.4 (7.35-7.45).
Composition: Sodium bicarbonate (NaHCO3) and Carbonic acid (H2CO3) in 20:1
 H++ NaHCO3 ↔ H2CO3 + Na+
 OH- + H2CO3 ↔ HCO3- + H2O

2). Phosphate Buffer System :

It is the major buffer system responsible for Intracellular pH maintenance-7.1 - 7.2
Composition: Mono hydrogen phosphate (HPO42-) and Di hydrogen phosphate (H2PO4-)
• H+ + HPO42- ↔ H2PO4-
• OH- + H2PO4- ↔ H2O + HPO42-

3). Protein Buffer System :

 Includes hemoglobin and works in blood.
 Proteins are made up of amino acids and having Carboxyl group(-COOH) and Amino group(-
 Carboxyl group gives up H+ where as Amino Group accepts H+.
 Side chains that can buffer H+ are present on 20 amino acids.
 Among all amino acids Hisitidine (H) and Cysteine (C) posses buffering capacity.
 The pH of the extracellular fluid of human body maintaining in the range of 7.35 – 7.45.

 The changes in pH leads to Acid-Base imbalance that may leads to the dysfunction of the Cellular
and metabolic activities.

 When the pH of blood is < 7.35, this condition referred as Acidosis.

 When the pH of the blood is >7.45, this condition referred as Alkalosis.

 Two types of conditions in Acidosis and Alkalosis, these include Respiratory & Metabolic.

ACIDOSIS : ( pH - <7.35 )
Respiratory Acidosis:
 Carbonic acid (H2CO3) levels increases; CO2 levels increases - > 45mmHg – Hypercapnea.
 It is due to Respiratory disorders- Pneumonia, Asthma.
Metabolic Acidosis:
 Bicrobonate (HCO3-) levels decreases (< 22 mEq/Litre) due to loss through Diarrhea and
Renal dysfunction.
 The main reason for this condition is untreated diabetes that leads to the production of
excess amount of ketone boides in body; these include β-Hydroxybutyric acid and
Aceto acetic acid. These ketone bodies are acidic in nature and reduces the pH of the blood
called Ketoacidosis which eventually leads to coma and finally death.
 Lacto acidosis – it due to excess of Lactic acid in blood due to strenuous exercise.

ALKALOSIS : ( pH - >7.45 )
Respiratory Alkalosis:
 Carbonic acid (H2CO3) levels increases decreases; CO2 levels decreases -< 35mmHg –
 It is due to hyperventilation and excess removal of CO2.
Metabolic Alkalosis:
 Bicarbonate (HCO3-) levels increases (> 26 mEq/Litre).
 It is due to excessive loss of acids due to prolonged vomiting.