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CHEMICAL BIOLOGY III

CHEM 403L

pH
and

Buffers
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Review of Terminologies

MOLARITY

Moles of solute
M=
Volume (L) of solution

Weight (g) of solute

=
(MWsolute)(L of solution)

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Review of Terminologies

LAW OF MASS ACTION

- At equilibrium, the product of the concentration of the product
divided by the product of the concentration of the reactants is a
constant, Keq.

wA + xB yC + zD

[C]y [D]z
Keq =
[A]w [B]x

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Review of Terminologies

Bronsted-Lowry Concept
Acids - release protons in their reactions

Bases - substances which accept protons

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Review of Terminologies

Acid-Base Conjugate Pairs

CH3CH2COOH + H2 O CH3CH2COO- + H3O+

acid base conjugate conjugate
base acid

NH3 + H2O NH4+ + OH-

base acid conjugate conjugate
acid base

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Acid-Base Conjugate Pairs

Exercise:
What are the conjugate bases of What are the conjugate acids of
these acids? these bases?

original conjugate original conjugate

acid base base acid
HNO3 OH-
H2O H2O
H3O+ HCO3-
H2SO4 SO42-
HBr ClO4-
HCO3-

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Strength of Acids and Bases

Strong acids - release protons readily and almost completely
in dilute aqueous solutions

Weak acids - at equilibrium less than 1% is ionized to yield

protons

Strong bases - have a great capacity for accepting protons

Weak bases - poor acceptor of protons

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ACIDS AND BASES

IONIZATION
STRONG ACIDS:
HCl H+ + Cl-
[HCl] = [H+]

e.g. 3 M HCl [H+] = 3 M

STRONG BASES:
NaOH Na+ + OH-
[NaOH] = [OH-]

e.g. 0.05 M NaOH [OH-] = 0.05 M

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ACIDS AND BASES

IONIZATION
WEAK ACIDS:
HA H+ + A-
[H+] [A-]
Keq = = Ka
[HA]

WEAK BASES:
BOH B+ + OH-
[B+] [OH-]
Keq = = Kb
[BOH]

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ION PRODUCT OF WATER

H2O H+ + OH-

[H+] [OH-]
Keq = = 1.8 X10-16
[H2O]
For [H2O] at room temperature: ρ = 1 g/ml = 1000 g/L
MW H O = 18 g/ml
2

ρ 1000 g/L
Dividing ρ by MW : = 55.6 moles/L
MW 18 g/mol

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ION PRODUCT OF WATER

[H+] [OH-]
Keq = = 1.8 X10-16
[H2O]

[H+] [OH-]
1.8 X10-16 = [55.6]

1 x 10-14 = [H+] [OH-] ; Kw = 1 x 10-14

Kw = [H+] [OH-]

Ion-product constant of H2O

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CONDITIONS
Neutral: [H+] = [OH-]

Acidic: [H+] > [OH-]

[H+] > 1.0 x 10-7 M; [OH-] < 1.0 x 10-7 M

Basic: [H+] < [OH-]

[OH-] > 1.0 x 10-7 M; [H+] < 1.0 x 10-7 M

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pH
- from French pouvoir hydrogène, “hydrogen power”

- defined as the negative logarithm of the concentration of H+ ions

pH = - log [H+]
[H+] = 10-pH = antilog (- pH)

CONDITIONS:
Neutral: pH = 7 NOTE: [H+] = [H3O+]
Acidic: pH < 7
Basic: pH > 7

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The pH scale

ACIDIC NEUTRAL BASIC

Decreasing [H+]
Increasing pH value

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pOH
pOH = -log [OH-]

[OH-] = 10-pOH = antilog (-pOH)

CONDITIONS:
pOH < 7.0 basic
pOH > 7.0 acidic
pOH = 7.0 neutral

RELATION:
pH + pOH = 14

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Measurement of pH

pH paper

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Measurement of pH

pH meter
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Average pH values of some body fluids

Gastric juice 1.4

Saliva 6.8
Urine 6.0
Milk 7.1
Tears 7.2
Blood 7.4
Pancreatic Juice 8.0

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CALCULATIONS
1. Calculate the pH of the following:
a. solution whose [H+] = 5 x10-4M.
b. 0.2 M HCl
c. 0.02 M NaOH
d. 0.0035 M CH3COOH, Ka = 1.8 x 10-5

2. Calculate the [H+] of a solution whose pH is 4.5.

3. Calculate the [OH-] of a solution whose pOH is 8.4.
4. Calculate the pOH of solution where the [H+] is equal to 5[OH-].
5. Calculate [OH-] in aqueous solution at 25OC when
[H+] = 0.01 mol L-.
6. Which of the following acids is stronger: boric
acid, pKa= 9.0, or acetic acid, pKa = 4.6.

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https://www.menti.com/

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Zwitterion

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BUFFER
-solution that resists pH changes

COMPONENTS:
• Weak acid + its conjugate base (salt)

• Weak base + its conjugate acid (salt)

NOTE:
A buffer solution which contains a weak acid and its salt is considered
to be acidic. One which contain a weak base and its salt is considered to
be a basic buffer.

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BUFFER
BUFFERING ACTION

A buffer solution is able to react with either H+ or OH- ions,

whichever is added. Thus, a buffer solution resists changes in pH.
When we add a modest amount or a strong base or a strong acid
to a buffer solution, the pH changes very little.

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BUFFER
CRITERIA FOR EFFECTIVE USE OF BUFFERS:
► the pH of the buffer should not be more than 1 pH unit above
or below the pKa
► the buffer should not have any metabolic effect with any
component of the assay medium
► should not react with any component of the assay medium
► prepare buffers in glassware instead of plastic ware
► prepare buffers at temperatures close to the working
temperature
► pH meters should be calibrated with at least two standard
buffered solutions
► corrections should be made for the influence of ionic strength
and temperature on pKa

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Common buffers in the laboratory

Acetate buffer (pH 3.6 to 5.6)

CH3COONa/CH3COOH
Phosphate buffer (pH 5.8 to 7.4)

Na2HPO4/NaH2PO4
Citrate buffer (pH 3.0 to 6.2)
Sodium Citrate/Citric acid
Barbitone buffer (pH 6.8 to 9.2)
Sodium diethyl barbiturate/Diethyl barbituric acid

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Blood buffers
1. Bicarbonate-carbonic acid (BHCO3 : H2CO3)

2. Hemoglobinate-hemoglobin (B.Hb : H.Hb)

3. Oxyhemoglobinate-oxyhemoglobin (B.HbO2 : H.HbO2)

4. Phosphate buffer (B2HPO4 : BH2PO4)

5. Protein buffer (B. Protein : H. Protein)

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HENDERSON-HASSELBALCH EQUATION

HA H+ + A-
[H+] [A-]
Ka =
[HA]
[HA]
Solving for [H+]: [H+] = Ka
[A-]
Taking negative logarithms of both sides:
[HA]
-log [H+] = -log Ka - log
[A-]

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

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Buffers in Relation to Acidosis & Alkalosis

Action of Lungs and Blood Buffers
1) in the presence of a weak acid

• if there is excess acid [H+], the conjugate base [HCO3-] reacts with the [H+] to form
the weak H2CO3, favoring an equilibrium shift to the left or backward reaction

• backward reaction decreases blood’s [H+], [HCO3-] and increases pCO2 in the lungs

• to remove excess CO2(g), the lungs then hyperventilate to maintain blood’s pH level

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Buffers in Relation to Acidosis & Alkalosis

Action of Lungs and Blood Buffers
2) in the presence of a weak base

• if there is excess base [OH-], the weak acid H2CO3 reacts with the [OH-] to form the
conjugate base [HCO3-], favoring an equilibrium shift to the right or forward reaction

• forward reaction decreases pCO2(g) in the lungs and increases blood’s [H+], [HCO3-]

• to conserve CO2(g), the lungs then hypoventilate to maintain blood’s pH level

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Buffers in Relation to Acidosis & Alkalosis

Acid-Base Balance of the Blood
Respiratory Acidosis: ↑[H+] = ↓pH

lungs cannot hyperventilate (hypoventilation - a symptom):

↑pCO2 ; ↑[HCO3-]

• excess CO2(g) cannot be efficiently removed due to lung disease (hypoventilation),

equilibrium shifts to the right, favoring forward reaction

• forward reaction increases *H++ (blood’s pH ≤ 7.35) and *HCO3-] resulting in

respiratory acidosis

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Buffers in Relation to Acidosis & Alkalosis

Acid-Base Balance of the Blood
Respiratory Alkalosis: ↓*H++ = ↑pH

lungs hyperventilate (hyperventilation - a symptom):

↓pCO2; ↓[HCO3-]

• CO2(g) is being removed rapidly by the lungs (hyperventilation), equilibrium

shifts to the left, favoring backward reaction

• backward reaction decreases *H++ (blood’s pH ≥ 7.35) and *HCO3-] resulting in

respiratory alkalosis

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Buffers in Relation to Acidosis & Alkalosis

Acid-Base Balance of the Blood
Metabolic Acidosis: ↑[H+] = ↓pH

lungs hyperventilate (a defense mechanism):

↓pCO2; ↓[HCO3-]

• *H++ increases (blood’s pH ≤ 7.35) thus, equilibrium shifts to the left, favoring
backward reaction

• backward reaction decreases blood’s *HCO3-] and increases pCO2 in the lungs

• to counteract metabolic acidosis, the lungs then hyperventilate to decrease pCO 2

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Buffers in Relation to Acidosis & Alkalosis

Acid-Base Balance of the Blood
Metabolic Alkalosis: ↓*H++ = ↑pH

lungs hypoventilate (a defense mechanism):

↑pCO2; ↑[HCO3-]

• *H++ decreases (blood’s pH ≥ 7.35) thus, equilibrium shifts to the right, favoring
forward reaction

• forward reaction decreases pCO2 in the lungs and increases blood’s *HCO3-]

• to counteract metabolic alkalosis, the lungs then hypoventilate to increase pCO2

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CALCULATIONS
1. Calculate the [H+] and the pH of a buffer solution that is 0.10 M in CH-
-5
3COOH (Ka = 1.8 x 10 ) and 0.20 M in CH3COONa.

2. Calculate the [OH-] and the pH of a solution that is 0.20 M aqueous NH3
and 0.10 M in NH4Cl.

3. If we add 0.010 mol of solid NaOH to 1.00 L of a buffer solution that is

0.100 M in CH3COOH and 0.100 M CH3COONa, how much will [H+] and pH
change? Assume that there is no volume change due to the addition of
solid NaOH.

4. A solution is 0.10 M in aqueous NH3. Calculate (a) the number of moles

and (b) the number of grams of NH4Cl that must be added to 500 mL of
this solution to prepare a buffer solution with pH 9.15. (neglect the
volume change due to addition of solid NH4Cl).

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HOMEWORK

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