Water, pH and dissociation of

water

DR.SADIA HAROON
 Lecture 3 Outline
• Homeostasis

• The structure and function of water

• Dissociation of weak acids and weak bases

• pH and the Henderson-Hasselbalch equation

• Buffers, biological/physiological examples

 HOMEOSTASIS
• The dynamic that defines the distribution of water and the
maintenance of pH and electrolyte concentrations
• Water distribution maintained by the kidneys, antidiuretic
hormone, hypothalamic thirst response, respiration and
perspiration
• Clinically, need to be aware of water depletion caused by
decreased intake (coma, wandering the desert) or increased
loss (diarrhea, renal malfunction, over-exercise), and
excess body water due to increased intake (too much I.V.)
or decreased excretion (renal failure)
 Importance of water
• 70% of earths surface is water
• Cell contains
• Water is present intra cellular and extra
cellularly
• Water is essential for life
• Chemical reaction takes place in aqueous
environment
 Water Chemistry
All living organisms are dependent on water.

The structure of water is the basis for its

unique properties.

The most important property of water is the

ability to form hydrogen bonds.
 Water Chemistry
Within a water molecule, the bonds between
oxygen and hydrogen are highly polar.

Partial electrical charges develop:

- oxygen is partially negative
- hydrogen is partially positive
 Water Chemistry
Hydrogen bonds are weak attractions between
the partially negative oxygen of one water
molecule and the partially positive hydrogen
of a different water molecule.

Hydrogen bonds can form between water

molecules or between water and another
charged molecule.
From Lehninger, 2nd ed., Ch 4
Hydrophobicity/Micelles
 Water Solubility / Hydrophilic

From Lehninger, 2nd ed., Ch 4

Water Chemistry
Structure of H20

From Lehninger, 2nd ed., Ch 4

 Water Chemistry
The polarity of water causes it to be cohesive and
adhesive.

cohesion: water molecules stick to other water

molecules by hydrogen bonding

adhesion: water molecules stick to other polar

molecules by hydrogen bonding
Water Chemistry
 Structure of water
• Water has dipolar structure
• V –shaped structure ,bend geometry
• Two light atoms; hydrogen
• One heavy atom oxygen
• H-O-H
• Angle between them109.47
 Why ice floats on water
• It expands on freezing
• It has density of 0.92g/ml
• Water at 0 degree has density of 1.0 g/ml
• Important in maintaining life and
enviornment.
 Structure of water
• Hydrogen bonding :non covalent bonding between
hydrogen and oxygen of two water molecules
• Vander walls forces
• Electrostatic interaction
• Hydrophobic interactions; results in formation of
droplet water water interaction)
 Other properties of water
• Thermal properties ;liquid at room temp
• At 0 degree melts ,at 100 degree boils
• Ice formation; maximum hydrogen
bonding
• Water vapour at 100 boiling
 Properties of Water
1. Water has a high specific heat.
- A large amount of energy is required to
change the temperature of water.

2. Water has a high heat of vaporization.

- The evaporation of water from a surface
causes cooling of that surface.
 Properties of Water
3. Solid water is less dense than liquid water.
- Bodies of water freeze from the top down.

4. Water is a good solvent.

- Water dissolves polar molecules and ions.
 Other properties
• Solvent properties
• Remarkable solvent
• Ionic substances and polar substances due to
hydrogen bonding
• Nonpolar lacks hydrogen bonding
• Amphipathic rearrange to form micelles;non
polar aggregate at centre non polar out side
 Properties of Water
1. Water has a high specific heat.
- A large amount of energy is required to
change the temperature of water.

2. Water has a high heat of vaporization.

- The evaporation of water from a surface
causes cooling of that surface.
From Lehninger, 2nd ed., Ch 4
Hydrophilic/Hydrophobic
Water Chemistry
Water Chemistry
Properties of Water
 Other properties
• Solvent properties
• Remarkable solvent
• Ionic substances and polar substances due to
hydrogen bonding
• Nonpolar lacks hydrogen bonding
• Amphipathic rearrange to form micelles;non
polar aggregate at centre non polar out side
 Ionization of water
• Water very less ionize
• [H+]+[OH-]
• [H+]+[H2O]=H3O=hydronium ion
• Hydrated proton
• Ke=ionization constant of water
• =[H+]+[OH-]/[H2O]
• (1.8x10-16)(55.5)=1.0x10-14
 Properties of Water
5. Water organizes nonpolar molecules.
- hydrophilic: “water-loving”
-hydrophobic: “water-fearing”
- Water causes hydrophobic molecules to
aggregate or assume specific shapes.

6. Water can form ions.

H2O  OH-1 + H+1
hydroxide ion hydrogen ion
 Ionization of water
• Water very less ionize
• [H+]+[OH-]
• [H+]+[H2O]=H3O=hydronium ion
• Hydrated proton
• Ke=ionization constant of water
• =[H+]+[OH-]/[H2O]
• (1.8x10-16)(55.5)=1.0x10-14
 pH =pH+pOH
• The product of both ion s as gram mole in one
liter of water is the ionic product of water.
• P= -log (logarithm) to base 10
• Log means ratio number
• Log of a number is the index by which 10 is to be
raised to get that number
 Ionization of water
• Water very less ionize
• [H+]+[OH-]
• [H+]+[H2O]=H3O=hydronium ion
• Hydrated proton
• Ke=ionization constant of water
• =[H+]+[OH-]/[H2O]
• (1.8x10-16)(55.5)=1.0x10-14
 Ph scale
• Kw=[H+][OH-]=10-14
• [H+]=[OH-]=10-7
• pH =-log [H+]
• PH= -log[10-7]
• P H =-[ -7]
• Ph =7
Function of Water:
Most of cellular activities are performed in
water solutions.
 4% TBW 40% TBW
Body Fluid

- makes up
~60% of total
body weight
(TBW)

- distributed in
three fluid
compartments.

16% TBW
 4% TBW 40% TBW

Fluid is continually
exchanged between
the three
compartments.

16% TBW
 4% TBW 40% TBW
Exchange between
Blood & Tissue Fluid

- determined by
four factors:
capillary blood
pressure
plasma colloid
osmotic
pressure
interstitium
Hydrostatic
Pressure
Interstitium
colloid osmotic
pressure
16% TBW
 4% TBW 40% TBW
Exchange between
Blood & Tissue Fluid
- not affected by
electrolyte
concentrations

- Edema = water
accumulation in
tissue fluid

16% TBW
Water Gain

Water is gained from

three sources.

1) food (~700 ml/day)

2) drink – voluntarily
controlled

3) metabolic water (200

ml/day) --- produced as a
byproduct of aerobic
respiration
Routes of water loss
1) Urine – obligatory (unavoidable) and
physiologically regulated, minimum 400
ml/day
2) Feces -- obligatory water loss, ~200 ml/day
3) Breath – obligatory water loss, ~300
ml/day
4) Cutaneous evaporation -- obligatory
water loss, ~400 ml/day
5) Sweat – for releasing heat, varies
significantly
ACID-BASE BALANCE
Acid
An acid is any chemical that releases H+ in
solution.

Base
A base is any chemical that accepts H+.
 Acids and Bases
Most biological buffers consist of a pair of
molecules, one an acid and one a base.
 Acids and Bases
Hydrogen ion (H+1 ) is the basis of the pH scale.

Greater H+1 concentration --- lower pH (acidic)

Lower H+1 concentration --- higher pH (basic)

pH
is the negative logarithm of H+ concentration,
and an indicator of acidity.

pH = - log [H+ ]

Example: [H+ ] = 0.1 µ M = 10 –7 M

pH
is the negative logarithm of H+ concentration,
and an indicator of acidity.

pH = - log [10 –7 ] = 7 log 10 = 7

Example: [H+ ] = 0.1 µ M = 10 –7 M

 pH
is the negative logarithm of H+ concentration,
and an indicator of acidity.

pH = - log [10 –8 ] = 8 log 10 = 8

Example: [H+ ] = 0.01 µ M = 10 –8 M

0.01 µ M [ H+ ] = pH 8 ↓ [ H+ ] = ↑ pH
0.1 µ M [ H+ ] = pH 7 ↑ [ H+ ] = ↓ pH
Normal functions of proteins (especially
enzymes) heavily depend on an optimal pH.

pH7.35-pH7.45
 Acids and Bases
Acid: a chemical that releases H+1 ions.

Base: a chemical that accepts H+1 ions.

Buffer: a chemical that accepts/releases H+1 as

necessary to keep pH constant
Regulation of acid-base balance

1) Chemical Buffers

2) Respiratory Control of pH

3) Renal Control of pH
 Properties of Water
1. Water has a high specific heat.
- A large amount of energy is required to
change the temperature of water.

2. Water has a high heat of vaporization.

- The evaporation of water from a surface
causes cooling of that surface.
 Properties of Water
3. Solid water is less dense than liquid water.
- Bodies of water freeze from the top down.

4. Water is a good solvent.

- Water dissolves polar molecules and ions.
 Acids and Bases
Hydrogen ion (H+1 ) is the basis of the pH scale.

Greater H+1 concentration --- lower pH (acidic)

Lower H+1 concentration --- higher pH (basic)

Acids and Bases
 Acids and Bases
Acid: a chemical that releases H+1 ions.

Base: a chemical that accepts H+1 ions.

Buffer: a chemical that accepts/releases H+1 as

necessary to keep pH constant
Acids and Bases
 WATER
• Comprises approx 70% of human mass (45-
60% intracellular, 25% extracellular/blood
plasma)

• dipolar: partial negative charge on oxygen,

partial positive charge on hydrogens

• dipolar nature leads to formation of many low

energy hydrogen bonds
 Water Solubility / Hydrophilic

From Lehninger, 2nd ed., Ch 4

From Lehninger, 2nd ed., Ch 4
Hydrophilic/Hydrophobic
From Lehninger, 2nd ed., Ch 4
Hydrophobicity
From Lehninger, 2nd ed., Ch 4
Hydrophobicity/Micelles
 Summary of water and pH
relationship
• Very low dissociation of H2O to H+ or OH-
• The ion product of H2O, Keq X 55.5 M, leads to
this: [H+] = [OH-] = 1 X 10-7 M for pure H2O
which is a constant in biological systems
• Therefore, if [H+] > 10-7 M, then [OH-] must be
less than 10-7 M, and vice versa.
• Thus, if the negative logarithm of [H+] is
derived ( pH = -log [H+] ), pure water would be
pH = 7, acids pH < 7, and bases pH > 7
 Ionization properties of water
• Water disociate
1) Polarity
Covalent bonds (electron pair is shared) between oxygen
and hydrogen atoms with a bond angle of 104.5o.

Oxygen atom is more electronegative that hydrogen atom

--> electrons spend more time around oxygen atom
than hydrogen atom --> result is a POLAR covalent
bond.

Creates a permanent dipole in the molecule.

Can determine relative solubility of molecules “like

 pH scale is widely used
in biological applications
^pH 7is neutral ,means no acidity or
alkalinity
^when excess of H+ ions are added the
solution are acidic
^when OH are added they become basic
^biomolecules have acidic or basic
properties
The Relationship Between pH and pOH

pH pOH [H+] mol/L [OH-] mol/L

0 14 1.0 10-14

2 12 0.01 10-12
4 10 0.0001 10-10
6 8 10-6 10-8
8 6 10-8 10-6
10 4 10-10 0.0001
12 2 10-12 0.01
14 0 10-14 1.0
 pH measurement
• pH paper:
1.Broad range (1-14whole no)
2.Narrow range (0.3-0.5pH unit)

• Universal indicator solution:

-It is a mixture of many indicators.
-Shows change of colour peculiar to the
composition of that mixture.

• Litmus paper:
-Its neutral point is pH 7.
-Blue colour on alkaline side and red colour on acid side.
 pH measurement
• pH meter:
-Electrometric method with

automatic temp compensating

device.
-Precision potentiometer.
-Controlled dial is calibrated in pH
units & milli volts
 pH measurement
• Calomel electrode:
-It is used as reference electrode
• Glass electrode:
-glass separates two solutions of different
H+ conc. A small potential arises across the
membrane this is propotional to the
difference in pH and can be measured by
amplifier.
 ACIDS AND BASES

• Acids are defined as hydrogen donor or proton donor.

• Bases are defined as hydrogen acceptor
• Strong acids and bases ionize completely in water.
• HCl=H+ Cl and Na OH=Na+ OH
• Strong acids are=sulfuric acid, nitric

Acid, hydrochloric acid and strong bases are

Na OH and KOH.
 ACIDS AND BASES

• Acids are defined as hydrogen donor or proton donor.

• Bases are defined as hydrogen acceptor
• Strong acids and bases ionize completely in water.
• HCl=H+ Cl and Na OH=Na+ OH
• Strong acids are=sulfuric acid, nitric

Acid, hydrochloric acid and strong bases are

Na OH and KOH.
 ACIDS AND BASES

• Acids are defined as hydrogen donor or proton donor.

• Bases are defined as hydrogen acceptor
• Strong acids and bases ionize completely in water.
• HCl=H+ Cl and Na OH=Na+ OH
• Strong acids are=sulfuric acid, nitric

Acid, hydrochloric acid and strong bases are

Na OH and KOH.
 Weak Acids and weak bases
• Weak acids partially ionize to release a
H+ ,and lower the pH.
• Weak bases accept a H+ and increase the
pH+
ACID BASE CONGUGATE PAIR
 pH of various common fluids

• Blood plasma 7.4

• Liver 6.9
• Muscle 6.1
• Saliva 6.6
• Urine 5-8
• Orange juice 4.3
• Vinegar 2.9
• Gastric juice 1.7
 Acids produce by the body
• Carbonic acid
• Sulphuric acid
• Phosphoric acid
• Lactic acid
• Citric acid
• Ammonium ion
• Ketone bodies
-Acetoactic acid
-Betahydroxy butyric acid
 Classification of acids
• First is based on strength:

1.Strong acids:
-That ionizes completely.
-High concentration of H ions.
-Free H+ show less tendency to
combine with base. (weakest conjugate)
2.Weak acids:
-Slowly dissociates.
-Give less no of H+ ions.
-Has the strongest conjugate base.
 Classification of acids
• Second is based on
volatile/non-volatile:

1.Volatile acid:
-Carbonic acid:20000 m eq/day
2.Non-volatile acid:
-Lactic acid
-Sulphuric acid
-phosphoric acid
 Classification of acids
• Second is based on
volatile/non-volatile:

1.Volatile acid:
-Carbonic acid:20000 m eq/day
2.Non-volatile acid:
-Lactic acid
-Sulphuric acid
-phosphoric acid
From Devlin, 3rd ed., Ch 1
Dissociation Constant and pH

From Marks, Marks, Smith, Ch 4

Henderson-Hasselbalch Equation
From Devlin, 3rd ed., Ch 1
From Devlin, 3rd ed., Ch 1
 From Devlin, 3rd ed., Ch 1
Sample pH problems
 From Devlin, 3rd ed., Ch 1
Sample pH Problem (cont)
 Buffers
• Definition: A weak acid plus its conjugate base
that cause a solution to resist changes in pH when
an acid or base are added

• Effectiveness of a buffer is determined by: 1) the

pH of the solution, buffers work best within 1 pH
unit of their pKa 2) the concentration
of the buffer; the more present, the greater the
buffering capacity
 Buffers and weak acids

• Buffers are solutions that resist change in

pH when acid or base is added to it.
• It consist of weak acid and its salt (acetic
acid and sodium acetate )or a weak base
and its salt (ammonium hydro oxide and
ammonium chloride )
 Mechanism of buffer action
• Added H+ ions = in the form of strong
acid ,combine with anions A-( largely form
the salt component of buffer), to form the
weakely dissociable HA, so that pH does
become as acid as it would be in the
absence of the buffer.
 Mechanism cont..
• Acetic acid freely ionizable
• Sodium acetate to a large extent
• CH3COOH=CH3COO+H+
• CH3COONa=CH3COO +Na+
• H Cl is added acetate will combine with
H+ to form acetic acid and NaCl is formed
 Mechanism contd..
• When NaOH is added the H+ of the
buffer (acetic acid) combine with OH- to
form water ,which is weakely
dissociated .
• Thus pH change due to base addition is
also prevented by buffer
• OH+H+ =H2O
 BUFFERING CAPACITY
• The efficiency of a buffer in maintaining a
constant pH on the addition of acid or base is
referred to as buffering capacity.
• The capacity to combine with added acid remains
so long as there is supply of the buffer salt in the
medium.
• OH can be buffered as long as some of the acid
HA remains to supply the H+.
 H-H EQUATION
• The quantitative relationship between the
concentrate of weak (HA) and its conjugate
(A-) is Henderson equation.
• HA=weak acid =H+ +A-
• H+ =proton
 H-H EQUATION
• The quantitative relationship between the
concentrate of weak (HA) and its conjugate
(A-) is Henderson equation.
• HA=weak acid =H+ +A-
• H+ =proton
 H-H EQUATION
• Is important for under standing buffer
action and acid –base balance in the blood
and tissue.
• Restating the expression for dissociation
constant of an acid
• The pH of a solution of a weak acid (or
base) and its salt is given by
• pH=pKa _log [HA/A-]
 H-H EQUATION
• Ke =equilibrium constant
• K e =ionization constant
• Or dissociation constant of an acid or Ka
• Stronger acids :phosphoric acid, carbonic
acid ,acetic acid have larger dissociation
constant
 Cont..
• Weaker acids mono hydrogen PO4 have
smaller dissociation constant
• Pka=log 1/Ka
• pKa =-logka
• The stronger the tendency to dissociate a
proton ,the stronger is the acid lower the
pKa
 H-H eq…
• Ka=[H+][A-]/HA
• First solve for [H+]=Ka [HA]/[A-]
• Take neg log on both side
-log [H+]=-log Ka –log [HA]/A-
• Putting values pH=pKa -log[HA]
• pH=pKa+log[A-]/[HA]
• pH=pKa
 Determination of pH
• Take three test tubes
• To one add 1ml sodium acetate+acetic
acid=(log 1/10)=-1
• 10 ml both(log 10/10)=0
• 10ml Na acetate +1ml acid (log10/1)=1
 Buffers of the body fluids
• Intracellular buffers;phosphate buffers
cossist of disodium hydrogen phosphate and
sodium dihydrogen phosphate.
• This has pka close to physiological ph
• Pka=6.8
 Contd.
• Protein buffers :depend on ionizable side
chain
• Histine imidazole group pka=6.1
• 16 histidine in albumin
• 38 histidine in haemoglobin
 Buffers …
• Extracellular ;comprise43%(intracellular 57%)
• 65%=bicarbonate buffers
• 30%=haemoglobin
• 4%=protein
• 1%=phosphate buffers
• Buffers are first line of defence against acid load
 Bicarbonate buffer system
• Consist of HCO3 and H2CO3
• Most significant
• 65% of plasma buffering
• 40% of buffering of body
• CO2 andHCO3 can diffuse easily across
membranes
• HCO3 (Metabolic component) regulated by
kidney and co2 by respiration.
 Buffers ..
• HCO3 22-26 mmol/lt (24mmol)
• Pka=6.1
• H2CO3=(CO2 in forms dissolved)
• pCO2=40
• Solubility coeffeciant=0.3
• pH=Pka+logHCO3/H2CO3
• 7.4=6.1+LOg24/1.2
• 7.4=6.1+LOg20
• 7.4=6.1+1.3
 Body buffers
• Three mechanism: to regulate pH and acid
base balance and maintain the blood pH
(around 7.4)
• 1.blood buffers
• 2.respiratory mechanism
• 3.renal mechanism
 EFFECTS OF ACID BASE
DISTURBANCES
• H+increased= acidosis,depression of C.N.S,
Disorientation, death in coma.
• H+decreased= alkalosis, overexcitability of
C.N.S,convulsions.
First in peripheral Nerves than C.N.S
Sensory effects,Tingling(pins and needle sensation
Motor effects, muscle twitches, spasm
Extreme alkalosis- spasm of respiratory muscles,
death
 EFFECTS…..
2. CHANGES IN ENZYMES ACTIVITY
Altering shape and activity of protein
molecule.
Some reactions are accelerated and some are
depressed.
 EFFECTS…..
3. CHANGES IN CELLULAR pH
Reduced contractility of actin and myosin in
muscles.
CHANGES IN POTASSIUM LEVEL
H+ ions enters the cells for sodium and potassium.
H+ ions are eliminated more than potassium so
hyperkalemia, cardic disfunction.
 Blood buffers
• Bicarbonate buffer system:
NaHCO/H2CO3
• H2CO3 >H+ +HCO3
• pH=pKa +log [salt]/[acid]
• 20:1 is the ratio
• Alkali reserve : responsible for effective
buffering of H+,generated in the body.
 Blood buffers.
• Bicarbonate buffers ;is index to understand the
disturbances.
• Phosphate buffers; this is important buffer
intracellular .
• Sodium di hydrogen phosphate and
disodium hydrogen phosphate
• Ratio of base to acid for phophate buffer is
4:1
• Pk is 6.8
 Body buffers
• Three mechanism: to regulate pH and acid
base balance and maintain the blood pH
(around 7.4)
• 1.blood buffers
• 2.respiratory mechanism
• 3.renal mechanism
 Physiological Buffers
• Carbon Dioxide-Bicarbonate System; a major regulator
of blood pH

• Phosphate System; major regulator of cytosolic pH

• [CO2] and [HCO3] are much higher than [PO4] in blood;

the reverse is true in the cytosol, [PO4] >>> [HCO3]
 Examples - Physiological Buffers

From Marks, Marks, Smith, Ch 4

From Marks, Marks, Smith, Ch 4
 pH Titration Curves

From Lehninger, 2nd ed., Ch 4

 Blood Bicarbonate and Metabolic
Acidosis
The bicarbonate blood buffer in a normal adult
maintains the blood pH at about 7.40. If the blood
pH drops below 7.35, the condition is referred to
as an ACIDOSIS. A prolonged blood pH below
7.0 can lead to death. Clinically for an acidosis,
the acid-base parameters (pH, [HCO3- ], [CO2] )
of the patients blood should be monitored. The
normal values for these are pH = 7.40;
[HCO3- ] = 24 mM; [CO2] = 1.2 mM.
 Sample Problem – Metabolic Acidosis

• The blood values of a patient were pH =

7.03 and [CO2] = 1.1 mM. What is the
patient’s blood [HCO3-] and how much of
the normal [HCO3-] has been used in
buffering the acid causing the
condition?

• The pK’ for [HCO3-]/[CO2] = 6.10

 Solution
• Substitute into Henderson-Hasselbalch equation:
• 7.03 = 6.10 + log [HCO3-]/1.1 mM, or
• 0.93 = log [HCO3-]/1.1 mM
• The anti-log of 0.93 = 8.5, thus:

• 8.5 = [HCO3 ]/1.1 mM, or [HCO3 ] = 9.4 mM

- -

• Since normal [HCO3 ] equals 24 mM, there was a decrease of 14.6 mmol of [HCO3
- -

per liter of blood in this patient. This would be approaching the point where, if left
untreated, the HCO3- buffering capacity would be no longer effective in this patient.