Aqueous Solutions:

Water&pH

and by
Drs. Selma Yilmaz and
Kamer Kilinc

Water: The solvent of life

Water on Earth moves continually
through the water cycle of evaporation
and transpiration (evapotranspiration),
condensation, precipitation, and runoff,
usually reaching the sea. Evaporation
and transpiration contribute to the
precipitation over land.

Aqueous Solutions: The cell

Most of the major components in cells (proteins, DNA and
polysaccharides) are also dissolved in water.

Water movement

b)

c)

d)

Biomedical Importance-I
Water is the predominant chemical component of
iving organisms and ideal biological solvent.
Water covers 71% of the Earth's surface. Only 2.5% of the Earth's water is

freshwater, and 98.8% of that water is in ice and groundwater. Less than
0.3% of all freshwater is in rivers, lakes, and the atmosphere, and an even
smaller amount of the Earth's freshwater (0.003%) is contained within
biological bodies and manufactured products
The total amount of water in a man of average weight (70 kilograms)
is approximately 40 litres, averaging 57 percent of his total body weight.
Intracellular fluid (2/3 of body water, extracellular fluid (1/3 of body
water)
In a newborn infant, this may be as high as 75 percent of the body
weight, but it progressively decreases from birth to old age, most of the
decrease occurring during the first 10 years of life.
Also, obesity decreases the percentage of water in the body,
sometimes to as low as 45 percent
ref.: Arthur Guyton 's Textbook of Medical Physiology

Biomedical Importance-II
Water is the chemical substances with
chemical formula H2O .
One molecule of water has two hydrogen
atoms covalently bonded to a single oxygen
atom.

Biomedical Importance-II
Waters peculiar physical properties,
including the ability to solvate a wide
range of organic and inorganic
molecules, derive from waters dipolar
structure and exceptional capacity for
forming hydrogen bonds.
The manner in which water interacts
with a solvated biomolecule
influences the structure of each.
An excellent nucleophile, water is
a reactant or product in
many reactions.

Biomedical Importance-III
Water has a slight tendency to
dissociate into
hydroxide ions and protons.
The acidity of a solution is generally reported using
logarithmic pH scale.
Bicarbonate and other buffers normally maintain the pH
of extracellular fluid between 7.35 and 7.45.
Suspected disturbances of acid-base balance are verified
by measuring the pH of arterial blood and the CO2
content of venous blood.

Biomedical Importance-V
Suspected disturbances of acid-base balance:
Acidosis and Alkalosis
Causes of acidosis
(blood pH <7.35)
include
diabetic ketosis
and lactic acidosis.
Alkalosis (pH>7.45)
may follow
vormiting of
acidic gastric
contents.
The main cause of
respiratory alkalosis
is
hyperventilation,
resulting in a loss
of carbon dioxide.

Soda lime

Biomedical Importance-VI
Regulation of water balance
(the flow of water in and out
of a system)
depends upon
the hypothalamic
mechanisms
on antidiuretic hormone
(ADH),
on retention or
excretion of water by
the kidneys, and
on evaporative loss.

Biomedical Importance-VIII
Diabetes insipidus (DI): A condition characterized by excessive thirst
and excretion of large amounts of severely diluted urine, with reduction
of fluid intake having no effect on the concentration of the urine.

Nephrogenic diabetes insipidus (NDI):

A form of diabetes insipidus,
is primarily due to kidney or
nephron dysfunction caused by
an insensitivity of the kidneys or
nephrons to ADH.

Biomedical Importance-VIII
Nephrogenic diabetes
insipidus (NDI) results
from the unresponsiveness
of renal tubular
osmoreceptors
to antidiuretic hormone,
leading to a decrease in
the ability of kidney to
concentrate urine by
removing free water or
adjust the subtle changes
in extracellular fluid
osmolarity.

Water Is An Ideal
Biological Solvent

Water molecule forms dipole.

Water molecule forms hydrogen
bonds.

Water Is An Ideal Biological Solvent

A water is an irregular, slightly skewed tetrahedron with an oxygen
at
its center.

The water molecule has a bent geometry with an O-H bond distance
of 0.958 and an H-O-H bond angle of 104.5o
Ideal tetrahedral angle is 109.5o.
The large electronegativity difference between H and O gives a 33%
ionic character on the O-H bond as is indicated by waters dipole
moment of 1.85 debye units.

Water Is An Ideal Biological Solvent

Water is a strong dipole
Water is dipole.
A water molecule is polar
because of the unequal
sharing of the electrons in
a bent structure.

Water Is An Ideal Biological Solvent

Water is a strong dipole
Water, a strong dipole,
has a high dielectric
constant.
The strongly
electronegative oxygen
atom pulls electrons away
from the hydrogen nuclei,
leaving them with a partial
positive charge, while its
two unshared electron pairs
constitute a region of
local negative charge.

Water has a high dielectric constant

As quantitatively described by Coulombs law,
the strength of interaction (F) between oppositely charged particles is
inversely proportionate to the electric constant () of the surrounding
medium.
Water has a high dielectric constant,
therefore greatly decreases
the force of attraction between
charged and polar species relative to
water-free environments with
lower dielectric constant.
Its strong dipole and high dielectric
constant enable water
to dissolve large quantities of
charged compounds such as salt.

Water molecules can form hydrogen bonds

Water has a slight tendency to dissociate into hydroxide ions
and protons.
The ability of water to ionize, while slight, is of central importance
for life.
Since water can act both as an acid and as a base, its ionization
may be represented as an intermolecular proton transfer that forms
a hydronium ion (H3O+) and a hydroxide ion (OH):

H2O+H2O = H3O+ + OH H3O+ (the hydronium ion) is abbreviated H; a bare proton has no
stable existence in aqueous solution.
The transferred proton is actually associated with a cluster of
water molecules.

Water molecules can form hydrogen bonds

A partially unshielded hydrogen
nucleus covalently bound to an
electron-withdrawing oxygen or
nitrogen atom can interact with
an unshared electron pair on
another oxygen or nitrogen
atom to form a hydrogen bond.

Since water molecules

contain both of these
features, hydrogen
bonding favors the selfassociation of water
molecules into ordered
arrays (Figure 22).

Structure of
water
.. ..
.. ..
.. ..

.. ..
.. ..

.. ..
.. ..

.. ..
.. ..

.. ..

Structure of Water

.. ..
.. ..

Water molecules can form hydrogen bonds

The boiling point of water (and all other liquids) is dependent on the
atmospheric pressure.
Mt. Everest water boils at 68 C, sea level water boils at 100 C at sea
level.
Water deep in the ocean near geothermal vents can reach temperatures
of hundreds of degrees and remain liquid.

Water molecules can form hydrogen bonds

Hydrogen bonding profoundly influences the physical properties of
water and accounts for its exceptionally high viscosity, surface tension,
and boiling point.
On average, each molecule in liquid water associates through hydrogen
bonds with 3.5 others.
These bonds are both relatively weak and transient, with a half-life of
one microsecond or less.
Rupture of a hydrogen bond in liquid water requires only about
4.5 kcal/mol, less than 5% of the energy required to rupture a
covalent OH bond.

Water molecules can form

hydrogen bonds
Hydrogen bonding enables water to dissolve many organic
biomolecules that contain functional groups which can
participate in hydrogen bonding.
The oxygen atoms of aldehydes, ketones, and amides,
for example, provide lone pairs of electrons that can serve
as hydrogen acceptors.

Water molecules can form hydrogen bonds

Alcohols and amines can serve both as hydrogen acceptors and
as donors of unshielded hydrogen atoms for formation of hydrogen
bonds (Figure 23).

Water molecules can form hydrogen bonds

Water as a Solvent
Pure water has a low electrical conductivity, but this increases with the
dissolution of a small amount of ionic material such as sodium chloride.
It is not only hydrogen bond acceptors or donors that dissolve well in
water. In contrast to most organic liquids, water is an excellent solvent for
ionic compounds. Substances like NaCl are very stable. They readily dissolve
in water. This is caused by dipolar nature of water. Dipoles interact with ions,
so cations and anions in aqueous solution are hydrated. They are surrounded
by shells of water molecules, called hydration shells.
High solubility of NaCl is caused by two factors:
1. The formation of hydration shells is energetically and thermodynamically
favorable.
2. High dielectric constant of water decreases the charge-charge interaction
between Na and Cl ions.
Substances that are readily dissolved in water are hydrophilic or waterloving substances.

Solvation of ions by oriented water molecules

Water as a Solvent

Nonpolar and nonionic compound does not dissolve in water.

Substances like aliphatic and aromatic hydrocarbons, therefore, are
called hydrophobic or water-fearing compounds.

A most interesting and important class of molecules are the ones that
exhibit both hydrophilic and hydrophobic properties simultaneously.
Such amphipathic compounds have a hydrophilic or polar head
group and a hydrophobic tail usually a hydrocarbon.

When one attempts to dissolve them in water, such compounds form

peculiar structures. They form a monolayer on the water surface, with
only the head groups immersed.

If the mixture is vigorously stirred, micelles (spherical structures

formed by a single layer of molecules) or bilayer vesicles may form. In
such cases the hydrocarbon tails lie in a roughly parallel array. In this
structure they interact via Van der Waals forces and hydrophobic
interactions. The polar or ionic heads are strongly hydrated. Most
important to biochemistry is the fact that amphipathic molecules form
the basis of the biological membrane.

Water as Solvent

INTERACTION WITH
WATER
INFLUENCES THE
STRUCTURE
OF BIOMOLECULES
Covalent & Noncovalent Bonds Stabilize
Biologic Molecules
Biomolecules Fold to Position Polar &
Charged Groups on Their Surfaces
Hydrophobic Interactions
Electrostatic Interactions
van der Waals Forces
Multiple Forces Stabilize Biomolecules

Covalent & Noncovalent Bonds Stabilize Biologic Molecules

1. The covalent bond is the strongest force that holds
molecules together.
2. Noncovalent forces, while of lesser magnitude, make
significant contributions to the structure, stability, and
functional competence of macromolecules in living cells.
These forces, which can be
either attractive or repulsive,
involve interactions both
within the biomolecule and
between it and
the water that forms
the principal component of
the surrounding environment.

Bond Energies for Atoms of Biologic Significance

Covalent Bonds Stabilize Biologic Molecules

The covalent bond is the strongest force that holds
molecules together.
A covalent bond is a chemical bond that involves the
sharing of electron pairs between atoms. The stable
balance of attractive and repulsive forces between atoms
when they share electrons is known as covalent bonding.

Covalent Bonds

Non-Covalent Interactions-1
Model

Type of interaction

1. Charge-Charge

2. Charge-Dipole

q+ q-

3. Dipole-Dipole

q+ q-

Dependence
of Energy on
Distance

Example

-NH3+

-OOC-

H2 O
1/r2

q+ q-

H2 O

H2 O

1/r3

-NH3+

1/r4

H2 O

1/r5

4.Charge-Induced
dipole

q+ q-

5.Dipole-Induced
dipole

q+

q-

q+

q-

+H

1/r

3N-

Non-Covalent Interactions-2
Model

Type of interaction

q+ q-

6. Dispersion

q-

q+

Dependence
of Energy on
Distance

Example

1/r6

7. Van der Waals interactions:

a. Attraction
b. Repulsion

1/r12

8. Hydrogen bonds
qAcceptor

q+
donor

C=O

H-N

Biomolecules Fold to Position Polar &

Charged Groups on Their Surfaces
Amphiphile (from the Greek amphis: both and philia: love,
friendship):
A chemical compound possesing both
hydrophilic (water-loving, polar, ionically charged) and
hydrophobic (lipophilic, fat-loving, nonpolar) properties.
Such a compound is called amphiphilic or amphipathic.
Common amphiphilic substances are soaps and detergents.
Most biomolecules are amphipathic; that is, they possess regions
rich in charged or polar functional groups as well as regions with
hydrophobic character.

Biomolecules Fold to Position

Polar & Charged Groups on Their Surfaces
Proteins tend to fold with the R-groups of amino acids with
hydrophobic side chains in the interior. Aminoacids with charged
or polar amino acid side chains (eg, arginine,glutamate, serine)
generally are present on the surface in contact with water.
A similar pattern prevails in a phospholipid bilayer, where the
charged head groups of phosphatidyl serine orphosphatidyl
ethanolamine contact water while their hydrophobic fatty acyl side
chains cluster together, excluding water.
This pattern maximizes the opportunities for the formation of
energetically favorable chargedipole, dipoledipole, and hydrogen
bonding interactions between polar groups on the biomolecule and
water.
It also minimizes energetically unfavorable contacts between water
and hydrophobic groups.

Amphiphiles Form Micelles and Bilayers

A Phospholipid

The most important reaction of amino acids is

the formation of a peptide bond (shaded).

Hydrophobic Interactions
Hydrophobic interaction refers to the
tendency of nonpolar compounds to
self-associate in an aqueous
environment.
Self-association minimizes energetically
unfavorable interactions between
nonpolar groups and water.
While the hydrogens of nonpolar groups
such as the methylene groups of hydrocarbons do not form hydrogen bonds,
they do affect the structure of the water
that surrounds them.

Hydrophobic Interactions

Water molecules adjacent to a hydrophobic group are restricted in

the number of orientations (degrees of freedom) that permit them
to participate in the maximum number of energetically favorable
hydrogen bonds.
Maximal formation of multiple hydrogen bonds can be maintained
only by increasing the order of the adjacent water molecules, with
an accompanying decrease in entropy.

Hydrophobic Interactions
It follows from the second law of thermodynamics,
the optimal free energy of a hydrocarbonwater mixture is a
function of both maximal enthalpy (from hydrogen bonding) and
minimum entropy (maximum degrees of freedom).
Thus,nonpolar molecules tend to form droplets in order to minimize
exposed surface area and reduce the number of water
molecules affected.
Similarly, in the aqueous environment of the living cell the hydrophobic
portions of biopolymers tend to be buried inside the structure of the
molecule, or within a lipid bilayer, minimizing contact with water.

Hydrophylic
groups

Fatty acid

WATER

Detergent

Phospholipid

WATER
Micelles
liposome

Electrostatic Interactions/Ionic Bonds

Interactions between charged groups help shape biomolecular structure.
Electrostatic interactions between oppositely charged groups within or
between biomolecules are termed salt bridges.
Salt bridges are comparable in strength to hydrogen bonds but act over
larger distances.

Electrostatic Interactions

Ionic interactions: Negatively charged

groups, such as the carboxylate group
(-COO-) in the side chain of aspartate
or glutamate, can interact with
positively charged groups, such as
the amino group (-NH3+) in the side
chain of lysine (see Figure 2.11)

Van der Waals Forces

van der Waals forces arise from attractions
between transient dipoles generated by the
rapid movement of electrons of all neutral
atoms. Significantly weaker than hydrogen
bonds but potentially extremely numerous,
van der Waals forces decrease as the sixth
power of the distance separating atoms.
Thus, they act over very short distances,
typically 24 .

Multiple Forces
Stabilize Biomolecules

Most of the major components

in cells (proteins, DNA and
polysaccharides) are also
dissolved in water.

The DNA double helix illustrates the contribution of multiple forces to the
structure of biomolecules..These noncovalent interactions include
hydrogen bonds between nucleotide bases (WatsonCrick base pairing)
and van der Waals interactions between the stacked purine and
pyrimidine bases.

Multiple Forces Stabilize

Biomolecules
The DNA double helix illustrates the
contribution of multiple forces to the
structure of biomolecules.

Multiple Forces Stabilize Biomolecule

Water is an excellent nucleophile

Metabolic reactions often involve the attack by lone pairs of
electrons residing on electron-rich molecules termed
nucleophiles upon electron-poor atoms called electrophiles.
Nucleophiles (nucleus-loving): electron-rich molecules,
nucleophiles donate electrons, Lewis bases, reacts with
positively (or partially positively) charged atoms
(electrophiles)
Electrophiles (electron-loving): electron-poor atoms,
positively charged atoms, electrophiles reacts with sources
of electrons (nucleophiles)

Water is an excellent nucleophile

Nucleophiles and electrophiles do not necessarily possess
a formal negative or positive charge.
Water, whose two lone pairs of sp3 electrons bear a partial
negative charge, is an excellent nucleophile.
Other nucleophiles of biologic importance include the
oxygen atoms of phosphates, alcohols, and carboxylic
acids; the sulfur of thiols; the nitrogen of amines; and the
imidazole ring of histidine.
Common electrophiles include the carbonyl carbons in
amides, esters, aldehydes, and ketones and the phosphorus
atoms of phosphoesters.

Water is an excellent nucleophile

Nucleophilic attack by water generally results in the cleavage of the
amide, glycoside, or ester bonds that hold biopolymers together.
Hydrolysis ( Greek: hydro-: "water",and lysis : "separation") usually
means the cleavage of chemical bonds by the addition of water.
This process is termed hydrolysis. Where a carbohydrate is broken into
its component sugar molecules by hydrolysis (e.g. sucrose being
broken down into glucose and fructose), this is termed saccharification.
Generally, hydrolysis or saccharification is a step in the degradation of a
substance.

Water is an excellent nucleophile

Conversely, when monomer units are joined together to
form biopolymers such as proteins or glycogen, water is a
product, for example, during the formation of a peptide bond
between two amino acids:

Water is an excellent nucleophile

In the cell, protein catalysts called enzymes

accelerate the rate of hydrolytic reactions
when needed.
Proteases catalyze the hydrolysis of proteins
into their component amino acids, while
nucleases catalyze the hydrolysis of the
phosphoester bonds in DNA and RNA.

Many Metabolic Reactions Involve

Group Transfer

D-G + A A-G + D

Many Metabolic Reactions

Involve Group Transfer
Given the nucleophilic character of water and its
high concentration in cells,
why are biopolymers such as proteins and DNA
relatively stable?
And how can synthesis of biopolymers occur in an
aqueous environment?
Central to both questions are the properties of
enzymes.

Many Metabolic Reactions

Involve Group Transfer
In the absence of enzymic catalysis, even
reactions that are highly favored
thermodynamically do not necessarily take
place rapidly.
Precise and differential control of enzyme
activity and the sequestration of enzymes
in specific organelles determine under what
physiologic conditions a given biopolymer
will be synthesized or degraded.
Newly synthesized biopolymers are not
immediately hydrolyzed, in part because
the active sites of biosynthetic enzymes
sequester substrates in an environment
from which water can be excluded.

WATER, ACIDS, BASES AND BUFFERS

WATER DISSOCIATES INTO HYDROXIDE IONS AND

PROTONS-1
Water has a slight tendency to transfer the proton that forms a hydronium
ion (H3O+) and a hydroxide ion (OH):
H2O+H2O = H3O+ + OH H3O+ (the hydronium ion) is abbreviated H; a bare proton has no stable
existence in aqueous solution.
The transferred proton is actually associated with a cluster of water
molecules.

WATER DISSOCIATES INTO HYDROXIDE IONS AND

PROTONS-1
At one instant WATER is an ion;
an instant later WATER is part of a water molecule.

WATER DISSOCIATES INTO HYDROXIDE IONS AND

PROTONS
For dissociation of water,r dissociation of water,

where the brackets represent molar concentrations (strictlyspeaking, molar

activities) and K is the dissociation constant.
Since 1 mole (mol) of water weighs 18 g,
1 liter (L) (1000 g) of water contains 1000 / 18 = 55.56 mol.
Pure water thus is 55.56 molar.

WATER DISSOCIATES INTO HYDROXIDE IONS AND

PROTONS
The molar concentration of H+ ions (or of OH ions) existence in pure
water
is the product of the probability (1.8 ~ 109 ) x the molar concentration of
water (55.56 mol/L ) = 1.0 ~ 107 mol/L.
The result is (1.8 ~ 109 ) x (55.56 mol/L ) = 1.0 ~ 107 mol/L.
We can now calculate K for pure water:

pH IS THE NEGATIVE LOG OF THE HYDROGEN ION CONCENTRATION

log10 X (10 to the X) is X = log (10X) = X

The term pH was introduced in 1909 by Sorensen, who defined

pH as the negative log of the hydrogen ion concentration:
pH = - log [H+]
This definition, while not rigorous, suffices for many biochemical
purposes.
To calculate the pH of a solution:
1. Calculate the hydrogen ion concentration [H+].
2. Calculate the base 10 logarithm of [H+].
3. pH is the negative of the value found in step 2.
For example, for pure water at 25 C, pH=-log 10-7 = 7
This value is also known as the power (English), puissant
(French), or potennz (German) of the exponent, hence the use
of the term p.

The pH scale
log10 (10 to the X) is X = log10 (10X) = X

log1/10=
log10-1= -1
log 0.1= -1
log10-2= -2

The following examples illustrate how to calculate the pH

of acidic and basic solutions.

The pH scale
Low pH values correspond to high
concentrations of H+ and high pH values
correspond to low concentrations of H+.

Solutions with a higher concentration of H+

than occurs in pure water have pH values
below 7 and are acidic. Solutions
containing molecules or ions that reduce
the concentration of H+ below that of pure
water have pH values above 7 and are
basic or alkaline.

pH
Low pH values correspond to high [ H+] and
high pH values correspond to low [ H+].
Acids are proton donors and bases are proton acceptors.
Strong acids (eg, HCl, H2SO4) completely dissociate into
anions and cations even in strongly acidic solutions (low pH).
Weak acids dissociate only partially in acidic solutions.
Similarly, strong bases (eg, KOH, NaOH)but not weak
bases (eg, Ca[OH]2)are completely dissociated at high pH.
Many biochemicals are weak acids.
Exceptions include phosphorylated intermediates, whose
phosphoryl group contains two dissociable protons, the first of
which is strongly acidic.

Minireview-1

Minireview-2

Minireview-3

The titration curve of acetic acid

Figure 1.3. The titration curve of acetic acid.
The molecular species that predominate
at low pH (acetic acid) and high pH (acetate)
are shown.
At low pH (high [H+]), the molecule is
protonated, and has zero charge.
As alkali added, [H+] decreases, (H+) and
(OH-) forms H2O,
acetic acid dissociates and loses its
proton, and the carbonyl group becomes
negatively charged.

Acids in the Blood of a Healthy Individual.

The Hendersen-Hasselbach Equation-1

The Hendersen-Hasselbach Equation-2

The Hendersen-Hasselbach Equation

log1/10=
log10-1=-1
log 0.1=-1
log10-2=-2

Dissociation of Weak Acids

CH3COOH

CH3COO- + H+

(HA)

Ka =

[H+] [A-]

-log

[HA]
[H+]

(A-)

[H+]

[HA]
= Ka
[A-]

= -log Ka - log

[A-]
pH = pKa + log
[HA]

[HA]
[A-]
Conjugated base
Acid

Henderson-Hasselbach Equation

Some weak acids or bases

acid

Conjugated base

H-COOH

CH3-COOH
CH3CHCOOH

H2CO3

N H4

3.75

CH3COO- +

4.76
3.86

OH
2.34
3.8

C6H5OH
+

HCOO- + H+

CH3CHCOO- + H+

OH
H3PO4

pK

H++H2PO4-

6.86

H+ + HCO3-

H++HPO4=
10.2

12.4

H+ PO43-

CO32- + H+

C6H5O + H+

9.89

NH3 + H+

9.25

BUFFERS-1
A buffer is an aqueous solution consisting of a mixture of a
weak acid and its conjugate base or a weak base and its
conjugate acid.
Its pH changes very little when a small amount of strong acid
or base is added to it and thus it is used to prevent changes in
the pH of a solution.
Buffer solutions are used as a means of keeping pH at a
nearly constant value in a wide variety of chemical applications.
Many life forms thrive only in a relatively small pH range so
they utilize a buffer solution to maintain a constant pH.
One example of a buffer solution found in nature is blood.

The Mechanism of Buffer Action

Because of reversible and partial dissociation, weak acids and their

conjugated base forms can act as proton donors and proton acceptors:

CH3COOH
NaOH

CH3COOH
HCl

CH3COO- + H+
Na+ + OH-

H2O

CH3COO- + H+
Cl- + H*

By the same mechanism, weak bases can act as buffers.

Figure 1.9. The titration curve of acetic

acid.
The molecular species that predominate
at low pH (acetic acid) and high pH
(acetate) are shown.
At low pH (high [H+]), the molecule is
protonated, and has zero charge.
As alkali added, [H+] decreases, (H+) and
(OH-) forms H2O,
acetic acid dissociates and loses its
proton, and the carbonyl group becomes
negatively charged.

BUFFERS

A buffer is any mechanism that resists changes in acidity (pH). The

body has two types of buffers:

Chemical buffers/ Physiological buffers - substances that bind

hydrogen ion and removes it from solution, as its concentration
begins to rise, and releases hydrogen ion as the concentration of a
solution begins to fall.

Physiological organ buffers - systems that control the bodys

output of acids, bases or carbon dioxide.

The four major chemical buffer systems of the body are the
bicarbonate, phosphate, hemoglobin and protein systems.

Major Chemical Buffers

in the biology/biochemistry laboratory
Acid form

pKa

Cacodylic acid

6.2

BISTRIS

6.5

PIPES

6.8

Imidazole

7.0

HEPES

7.6

Tris

8.3

Major Chemical Buffers in Human Body/

Physiological Buffers
Buffer

Conjugate
Acid

Conjugate Base

Main buffering Site

Hemoglobin HHb

Hb-

erythrocytes

Proteins

HProt

Prot-

Intracellular,
extracellular

Phosphate

H2PO4-

HPO42-

Intracellular

HCO3-

Extracellular

Bicarbonate CO2

H2CO3-

Chemical Buffers in Human Body/

Physiological Buffers

1. Hemoglobin
2. Proteins

Non-bicarbonate
buffers

3. Phosphate buffer system

4. Carbonic acid / Bicarbonate system

Carbonic acid-Bicarbonate Buffer system

H2CO3
HCO3-

H+ + HCO3H+ + CO32-

CO2 + H2O

H2CO3

CA

pK(apperent) =

[HCO3-] [H+]
H2CO3

7.4 = 6.1 + log

pK1 = 3.8
pK2 = 10.2
H+ + HCO3-

= 6.1
[HCO3-]
[H2CO3]

HCO3- / H2CO3 ratio is 20/1

Phosphate Buffer System

H2PO4-

pK = 6.8

H+ + HPO42-

2HPO
4
7.4 = 6.8 + log
H2PO4-

HPO42- / H2PO4 = 4/1

* Major intracellular inorganic buffer.
*H2PO4 excretion in urine is important for
the regulation of blood pH.

Plasma
Tissue
metabolism

CO2
CO2

Red blood cells

CO2 + H2O

H2CO3

H2 O

H2CO3 HCO3Cl-

HCO3- + H+
Cl-

H+ + HCO3-

HbO2

HHb
O2

CA

O2

release

O2

Carbamino Hb

HPO4=

H2PO4-

Plasma

Alveols

CO2

CO2

Red blood cells

CO2 + H2O

H2CO3

H2 O

H2CO3 HCO3Cl-

HCO3- + H+
Cl-

H+ + HCO3-

HbO2

HHb
O2

CA

O2

diffusion

O2

Carbamino Hb

HPO4=

H2PO4-

Hydrochloric Acid

Hydrochloric acid (HCl), also called gastric acid, is secreted by

parietal cells of the stomach into the stomach lumen, where the
strong acidity denatures ingested proteins so they can be degraded
by digestive enzymes.

When the stomach contents are released into the lumen of the small
intestine, gastric acid is neutralized by bicarbonate secreted from
pancreatic cells and by cells in the intestinal lining.

Effects of pH on Proteins
Proteins can be either positively or negatively charged based on the pH of
the surrounding solution. Amino acids that make up proteins may be positive,
negative, neutral, or polar in nature, and together give a protein its overall
charge.
At a pH below their pI (isoelectric point), proteins carry a net positive charge;
at a pH above their pI, they carry a net negative charge.

Effects of pH on Proteins

Effects of pH on Proteins
Protein denaturation
If there is an increase in the pH of the surrounding solution and it becomes too
basic, the amino acid bonds will break and the protein will be denatured. The
same is true if there's increased acidity.

Clinical Causes of Acid-Base Disorders

Metabolic acidosis

Respiratory acidosis

Metabolic alkalosis

Respiratory alkalosis

Diabetes mellitus
(ketoacidosis)

Chronic obstructive
airways disease

Vomiting (loss of
hydrogen ion)

Hyperventilation (anxiety,
fever)

Lactic acidosis

Severe asthma

Nasogastric suction

Lung diseases
associated with
hyperventilation

Renal failure (inorganic

acids)

Cardiac arrest

hypokalemia

Anemia

Severe diarrhea (loss of

bicarbonate)

Depression of respiratory
center (drugs, opiates)

i.v. Administration of
bicarbonate (after cardiac
arrest)

Salicylate poisoning

Renal and intestinal loss

of HCO3-

Weaknes of respiratory
muscles

Impairment of renal H+
excretion