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pH Scale, Buffers,

Redox potential
Ifra
M.Sc. 3rd
SEM
pH Scale
Danish biochemist ‘ Soren Sorensen’ used a logarithmic scale for
expressing the hydrogen ion concentration. This scale was called pH
scale, where p stands for power and H for Hydrogen ion
concentration. He defined pH as
“The pH of a solution is the negative logarithm of the concentration(in
moles/litre) of hydrogen ions”
pH= ─ log [H+] or log 1/[H+]
pH gives an idea about the acidity or basicity of the solution. The
ionization product of water form the basis of pH scale.
Since pure water is neutral, it contains equal concentration of
hydrogen & hydroxyl ions. At a certain temperature, the product of
the concentration of H+ & OH+ ions in pure water is known as ionic
product of water at the temperature. The ionic product of water at
25°C is approx. equal to 1*10-14

In neutral solution [H+]=[OH-]= 10-7 (pH = 7).

In acidic solution, pH value ranges from 0 to 7.
In alkaline(basic) solution, the pH value is between 7 to 14
A pH scale shows a range of 0 to 14.
Sometimes the expression pOH is used to denote the basicity of OH-
concentration of a solution.
pH=log 1/[OH-]= ─log [OH-]
pH + pOH = 14
The pH of an aqueous solution can be estimated by using various
indicator, dyes such as phenolphthalein and phenol red. These dyes
ionize at a specific pH to produce coloured ions. In laboratories, the
pH meter.
pH OF SOME FLUIDS

FLUID pH
1. Gastric juice 1.0
2. Lemon juice 2.0
3. Tomato juice 4.0
4. Milk , Saliva 6.5
5. Human blood 7.4 – 7.8
BUFFERS

SOLUTIONS OF RESERVE
ACIDITY AND RESERVE
ALKALINITY
BUFFERS
A buffer solution is that which tends to maintain its pH when small
amounts of strong acid or base are added to it. It is a mixture of weak
acid or weak base and their conjugate base or conjugate acid
respectively. It contains a hydrogen ion donor & a hydrogen ion
acceptor form of a weak acids and weak bases. A buffer system is
most effective when the concentration of H+ & H+ acceptor is equal.
Carbonic acid bicarbonate is a common buffering system in blood
plasma. The weak carbonic acid dissociated into H+ & HCO3- as follows-
H2CO3 → H+ + HCO3-
When a small amount of HCl is added to this system, H+ ions are
produce from the acid combine HCO3- ions to form H2CO3. If a small
amount of NaOH is added, the OH produced reacts with H+to form
water molecules. Thus this system soaks the H+ or OH- produced
from strong acid or base & tend to maintain the original pH.
Similarly weak bases & their salts also work as buffer system.
Buffer systems in the organism help in carrying on most of the
biochemical reactions in a narrow pH range of 6 to 8. The blood for
example maintains its constant pH of about 7.4 despite the fact that
it carries a large number & variety of chemicals. Buffer system
provide protection to cells & tissues against sudden change in pH.
Buffering in the blood
The pH range of blood is normally in the range of 7.35 to 7.45. If pH
decreases below this range, the symptoms of acidosis appear & death
of the animal may occur at pH 7.8 . This is because the enzymes
present in the blood are extremely sensitive to changes in pH. The
major buffer systems of the blood are bicarbonate, phosphate,
haemoglobin & proteins buffers. Haemoglobin is a good buffer
because of its capacity to act as a oxygen acceptor as well as a oxygen
donor. Oxyhaemoglobin( HHbO2) is a stronger acid than than
carbonic acid but haemoglobin(HHb) is weaker acid. When blood is
circulated through the pulmonary veins( in lungs), haemoglobin is
converted to oxyhaemoglobin by absorbing oxygen. Because of its
acidic nature it reacts with the bicarbonates present in blood.
HHbO2 + BHCO3 → BHbO2 + H2CO3 ( B= Na, K, etc)
The carbonic acid thus produced is decomposed into carbon dioxide &
water by the enzyme carbonic anhydrase.
H2CO3 → CO2 + H2O
The salt of oxyhaemoglobin formed during reaction with bicarbonate
converted to the salt of haemoglobin by deoxygenation in the tissues,
which then reacts with the carbonic acid produced from carbon dioxide
liberated in the oxidation of carbohydrates, to form bicarbonate salt
and haemoglobin.
BHbO2 → BHb + O2
BHb + H2CO3 → BHCO3 + HHb
The haemoglobin thus liberated goes to the lungs again, where it can be
oxygenated. This cyclic oxidation and deoxygenation of haemoglobin
between lungs and tissues is represented as Henderson cycle.
Haemoglobin as buffer

PLASMA ERYTHROCYTES

CO2 CO2 +H2O

Carbonic anhydrase
H2CO3
HHb

HCO3- HCO3- + H+
Hb
Cl- Cl-
Renal regulation of blood pH
PHOSPHATE BUFFER
BLOOD Renal tubular cell TUBULAR LUMEN
Na2HPO4 pH=7.4
Na+ Na+
Na+ NaHPO4-
HCO3- HCO3- + H+
H+

H2CO3
NaH2PO4 pH=4.5
CO2 + H2O

EXCRETED
PROTIEN BUFFER SYSTEM

BLOOD RENAL TUBULAR CELL TUBULAR LUMEN

Glutamine

Glutaminase
NH3
Glutamate
Na+
Na+ Na+
H+
HCO3- HCO3- + H+
CA
NH4+
H2CO3
CO2 + H2O
EXCRETED
pH Regulation In Blood
pH=7.4

CO2 Lungs
Metabolism
(H2CO3) (CO2 exhaled)
(CO2 generated)

HCO3-

Erythrocytes
(CO2 Kidneys
transported , (HCO3-
HCO3- generated ,H* lost)
generated)
DISEASES OCCUR DUE TO pH
DISTURBANCE
ACIDOSIS – It is a decline in ALKALOSIS – It is a rise in
pH pH
(a)METABOLIC- due to decrease in (a)METABOLIC- due to increase in
bicarbonate. bicarbonate.
(b)RESPIRATORY- due to an increase (b)RESPIRATORY- due to decrease in
in carbonic acid . carbonic acid.
(c)Occur due to diabetes, heart, liver, (c)Occur due to vomiting, anemia,
lungs and kidney problems. hypokalemia and at high altitude.
(d)Compensated by hyperventilation (d)Compensated by hypoventilation
and HCO3- retained by kidney. and HCO3- excretion by kidney.
Other Examples
Buffering Agent pKa Useful pH Range
CITRIC ACID 3.13,4.76,6.40 2.1- 7.4
ACETIC ACID 4.8 3.8-5.8
KH2PO4 7.2 6.2-8.2
CHES 9.3 8.3-10.3
BORATE 9.24 8.25-10.25
USES
• In determining pHof unknown solutions.
• In studying the rate of chemical reactions.
• In the manufacture of ethyl alcohol from molasses (pH 5-6.8).
• In paper manufacture , leather tanning etc.
• In preparing cultures in biological specimens.
pH of the buffer system
pH of the buffer solution can be calculated if the
composition of the mixture as well as the ionization constant
of the weak electrolyte is known. For example in a buffer
mixture of acetic acid and sodium acetate, the pH can be
determined if the ionization constant of acetic acid is
known. The formula for such a determination can be
derived as follows-
Hendorson-Hassel Equation
1. For acid , pH=pKa + (salt)/(acid)
2. For base , pOH=pKb + (salt)/(base)
3. pH + pOH= pKw =14
4. In blood, pH=pKa +log(base)/(acid)= pKa+log(HCO3-)/(H2CO3)

H2CO3 H+ + HCO3-
REDOX POTENTIAL
The process of electron transfer accompanied with the
oxidation – reduction of the system. A compound losing
electron is oxidized while a compound gaining it is reduced.
For example ferrous ion is oxidized to ferric ion is oxidized to
ferric ion by losing one electron & vice-versa. The
quantitative measure of the affinity of a compound to lose or
gain electron is the redox potential.
Fe++ → Fe+++ + e-
A redox system can be compared to a dry electric cell. In a cell,
the e- are transferred through a wire from one electrode to the
other. This generates an electric an electric current. The
capacity to gain or lose e- in such a system is electrode
potential. It can be measured through a standard hydrogen
potential of zero, at 1N concentration & 1 atmospheric pressure.
The electrode potential of a reducing- oxidizing system can also
measured in a similar way. Electrode potential in this case will
be the redox potential.
Redox potential of an organic compound can be measured in the
laboratory by using a standard platinum electrode.
E = E• + RT/nF In [oxidant]/[reductant]
where E= redox potential
E•= Redox potential of mixture containing equimolal
concentration of oxidant and reductant.
R= Gas constant
T= absolute temperature
n= number of e-
This equation is called Peter’s equation.
Under normal conditions of temperature i.e 30◦C, valency
change of 2 & converting into log10 [oxidant]/[reductant]
Redox potential of hydrogen involving system
E=E• + RT/nF In[oxidant]/[reductant]+ RT/nF
In[H+]

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