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Water, pH and Buffers

Textbook: Biochemistry by Campbell and Farrell 8th


ed CENGAGE Learning
What makes water polar
What is a polar bond:
Electrons are unequally shared,more negative charge found closer to one
atom.
Due to difference in electronegativity of atoms involved in bond.
Electronegativity
Electronegativity: a measure of the force of an
atoms attraction for electrons it shares in a chemical
bond with another atom
Oxygen and Nitrogen, more electronegative than
carbon and hydrogen
Fluorine is most electronegative (4)
Polar Bonds & Molecules
Molecules such as CO2 have polar bonds but, given
their geometry, are nonpolar molecules; that is, they
have a zero dipole moments
Solvent Properties of H2O

Ionic compounds (e.g.,KCl) and low-molecular-


weight polar covalent compounds (e.g., C2H5OH and
CH3COCH3) tend to dissolve in water
The underlying principle is electrostatic attraction of
unlike charges; the positive dipole of water for the
negative dipole of another molecule, etc.
ion-dipole interaction: e.g., KCl dissolved in H2O
dipole-dipole interactions: e.g., ethanol or acetone
dissolved in H2O
dipole induced-dipole interactions: weak and generally
do not lead to solubility in water
Hydration Shells Surrounding Ions in Water
Ion-dipole and Dipole-dipole Interactions

Ion-dipole and dipole-dipole interactions help


ionic and polar compounds dissolve in water
Solvent Properties of H2O
Hydrophilic: water-loving
tend to dissolve in water
Hydrophobic: water-fearing
tend not to dissolve in water
Amphipathic: has characteristics of both properties
molecules that contain one or more hydrophobic and
one or more hydrophilic regions, e.g., sodium palmitate
Amphipathic molecules
both polar and nonpolar character
Interaction between nonpolar molecules is very weak
called van der Waals interactions
Micelle formation by amphipathic molecules
Micelle: a spherical arrangement of organic
molecules in water solution clustered so that
their hydrophobic parts are buried inside the sphere
their hydrophilic parts are on the surface of the sphere
and in contact with the water environment
formation depends on the attraction between
temporary induced dipoles
Examples of Hydrophobic and Hydrophilic
Substances
Hydrogen Bonds
Hydrogen bond: the attractive interaction between
dipoles when:
positive end of one dipole is a hydrogen atom bonded
to an atom of high electronegativity, most commonly O
or N, and
the negative end of the other dipole is an atom with a
lone pair of electrons, most commonly O or N
Hydrogen bond is non-covalent
Interesting and Unique Properties of Water
Each water molecule can be involved in 4 hydrogen
bonds: 2 as donor, and 2 as acceptor
Due to the tetrahedral arrangement of the water
molecule (Refer to Figure 2.1).
Hydrogen Bonding
Even though hydrogen bonds are weaker than
covalent bonds, they have a significant effect on the
physical properties of hydrogen-bonded compounds
Some Physical Properties of Water and Other Compounds.
Substance Melting Boiling Heat of Heat Heat of
Point, C Point, C vaporization, capacity, fusion, cal/g
cal/g cal/g

Water 0 100 540 1.000 80

Ethanol -114 78 204 0.581 24.9

Methanol -98 65 263 0.600 22

Acetone -95 56 125 0.528 23

Ethyl acetate -84 77 102 0.459 -

Chloroform -63 61 59 0.226 -

NH3 -78 -33 327 1.120 84

H2S -83 -60 132 - 16.7

HF -92 19 360 - 54.7


Comparison
between the H-
bonding of water
and ethyl alcohol.
There are 4 water
molecules bonded
to one molecule of
water as opposed
to only 3 EtOH
molecules per
EtOH molecule.
Water passes through a maximum
density of 1 g/mL at 4C.

This explains why ice floats because at


freezing temperatures, ice is less dense than
liquid water.
Other Biologically Important Hydrogen bonds

Hydrogen bonding is important in stabilization of 3-D


structures of biological molecules such as: DNA,
RNA, proteins.
Acids, Bases and pH
Acid: a molecule that behaves as a proton donor

Strong base: a molecule that behaves as a proton


acceptor
Acid Strength
One can derive a numerical value for the strength of an
acid (amount of hydrogen ion released when a given
amount of acid is dissolved in water).
Describe by Ka:

Written correctly,
Ionization of H2O and pH
Lets quantitatively examine the dissociation of water:

Molar concentration of water (55M)

Kw is called the ion product constant for water.

Must define a quantity to express hydrogen ion concentrationspH


Henderson-Hasselbalch
Equation to connect Ka to pH of solution containing
both acid and base.
We can calculate the ratio of
weak acid, HA, to its conjugate
base, A-, in the following way
The law of mass action
aA + bB cC + dD

The law states that at equilibrium: the product of


the concentrations of the substances formed in a
chemical reaction divided by the product of the
concentrations of the reactants in that reaction is
a constant known as the equilibrium constant,
Keq.
Keq and Kw

Ion product
of water
Keq and Ka

HA + H2O H3O+ + A-
Example:

Acid dissociation
constant
Bronsted-Lowry acid-base concept:
Acid: proton donor (H+) Base: proton acceptor

HA + H2O H3O+ + A-
HA is the acid, H2O is the base

H2O + NH3 NH4+ + OH-


H2O is the acid, NH3 is the base
HA + H2O H3O+ + A-
HA is the Bronsted acid, H2O is the Bronsted base

Conjugate acid-base pair: related through the loss


or gain of a proton

The conjugate acid-base pairs above are:


HA A- and H3O+ - H2O
Kw = [H+] [OH-] = 1.0 x 10-14
1.0 x 10-14 = (1.0 x 10-7) (1.0 x 10-7)

Sorensen and pH:

pH = -log [H+] pOH = -log [OH-]


pKw = pH + pOH
14 = 7 + 7
The dissociation of a weak acid, e.g. CH3COOH, in water:

HA + H2O H3O+ + A-

(the Henderson-Hasselbach equation, the


equation to use in preparing buffer solutions)
Buffer solution: a solution that resists a change in pH
on the addition of acid or alkali

Composition: weak acid and its conjugate base


weak base and its conjugate acid

Factors that affect effectivity (or capacity) of buffers:


1. Concentration (sum of the concentrations of the
weak acid and its conjugate base); 0.1M or
higher is best
2. Ratio of the concentration of the conjugate base
and the concentration of the weak acid.
buffer range: pH = pKa + 1
Buffer Capacity
Buffer capacity is related to the concentrations of
the weak acid and its conjugate base
the greater the concentration of the weak acid and its
conjugate base, the greater the buffer capacity
Naturally Occurring Buffers
H2PO4-/HPO42- is the principal buffer in cells
H2CO3/HCO3- is an important (but not the only) buffer
in blood

hyperventilation can result in increased blood pH


hypoventilation can result in decreased blood pH
(Biochemical Connections p. 60)
Other component buffers in circulating blood:
1. Oxygenated and non-oxygenated forms of
hemoglobin
2. Plasma proteins
pH range of blood: 7.35 -7.45
best buffer: pH = pKa
Q: Is the H2CO3-HCO3- buffer system a good buffer?
Does it satisfy the pH = pKa?

The H2CO3 has a pKa of 6.1 which would mean that the
ratio is 1H2CO3: 20 HCO3-, pKa pH. But because of the
equilibrium
H2CO3 CO2 gas + H2O
the ratio 20:1 is maintained by respiration by peripheral
cells or expiration in the lungs. In other words, there is no
depletion of the buffer components.
Laboratory Buffers
Experiment 1
pH and BUFFER SOLUTIONS
A. Measurement of pH

Nine beakers at 0.1 M each of the following


solutions:
HCl, CH3COOH, NaC2H3O2, NaH2PO4,
Na2HPO4, NH3, NaOH, Tap water and Distilled
water.

Take the pH of each using pH paper and a pH


meter.
B. pH of the Buffer Systems
Get six 50 mL beaker. Label it as A, B, C, D, E, F. Prepare buffer
systems in six beakers as follows:
A. 10 mL 0.1 M CH3COOH + 10 mL 0.1M NaC2H3O2
B. 2 mL 0.1M CH3COOH + 20 mL 0.1 M NaC2H3O2
C. 20 mL 0.1M CH3COOH + 2 mL 0.1 M NaC2H3O2
D. 10 mL 0.1M NaH2PO4 + 10 mL 0.1M Na2HPO4
E. 2 mL 0.1M NaH2PO4 + 20 mL 0.1M Na2HPO4
F. 20 mL 0.1M NaH2PO4 + 2mL 0.1 M Na2HPO4

1. Determine the pH of each buffer using pH paper.


2. Divide contents of beaker into two by placing the
other half in a beaker or test tube.
3. To one portion, add 1 mL of 0.1 M HCl. Determine
the pH using pH paper.
4. To the other portion, add 1 mL of 0.1 M NaOH.
Determine the pH using pH paper.

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