08.3502/3-002 INTERMEDIATE BIOCHEMISTRY I Sept.

, 14, 2005

SOLUTIONS to NUMERICAL PROBLEMS in Self-Test Exercises #1

1. Given that the pKa of acetic acid is 4.76, calculate the volumes of 2.00 M acetic acid and 1.00
M NaOH required for preparing 500 ml of a 0.10 M acetate buffer with a pH of 4.80.
(Ans. = 25.00 ml of 2.00 M acetic acid; 26.15 ml 1.00 M NaOH)

Solution:
The relevant weak acid dissociation is: CH3 COOH + H2 O » CH3 COO- + H3 +O

Note that CH3 COO- present in the buffer (you will make) is formed from CH3 COOH by reaction with the
strong base NaOH in a 1:1 stoichiometry:

CH3 COOH + NaOH º CH3 COO- + Na+ + H2 O

pH = pKa + log {[CH3 COO-]/[CH3 COOH]}

4.8 = 4.76 + log {[CH3 COO-]/[CH3 COOH]}
4.8-4.76 = log {[CH3 COO-]/[CH3 COOH]}
0.04 = log {[CH3 COO-]/[CH3 COOH]}
100.04 = [CH3 COO-]/[CH3 COOH]
1.0965/1 = [CH3 COO-]/[CH3 COOH]

Alternative One Approach for calculating # of mols of each buffer component from [PA]/[PD] ratio:
Thus, Fraction of buffer consisting of CH3 COO- = 1.0965/(1 + 1.0965) = 0.523
ˆ Fraction of buffer consisting of CH3 COOH = 1 - 0.523 = 0.477

# moles (CH3 COO- + CH3 COOH) in 1 litre buffer = 0.1 mol per litre buffer [= 0.10 M]

ˆ# moles (CH3 COO- + CH3 COOH) in 500 ml buffer = (0.1 mol. L-1 )(0.5L)
= 0.05 mol in 500 ml buffer

# moles CH3 COO- in the 500 ml buffer = (0.05 mol)(0.523)

= 0.02615 mol CH3 COO- in 500 ml buffer.

The stoichiometry dictates that you would require 0.02615 mol NaOH to produce 0.02615 mol CH3 COO-
from 0.02615 mol CH3 COOH.

Vol of 1.00M NaOH to contain 0.02615 mol NaOH:

1.00M NaOH contains 1 mol/L [Meaning 1L/mol]
ˆ0.02615 mol NaOH will be contained in (1L/mol)(0.02615mol) = 0.02615 L
=26.15 ml of 1.00 M NaOH

CH3 COOH Needed:

Moles CH3 COOH present in 500 ml 0.10M acetate buffer pH 4.8
= (0.05 mol)(0.477) = 0.02385 mol CH3 COOH in the 500 ml buffer

Thus, total # moles CH3 COOH required to make 500 ml 0.10 M acetate buffer pH 4.8
= Total # moles of buffer components in the 500 ml
= (0.02616 mol + 0.02385 mol) = 0.05 mol

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Stock CH3 COOH solution = 2 M [i.e., 2 mol/L]
Thus, 0.05 mol CH3 COOH is contained in (0.05 L/2) = 0.025 L
= 25 ml of 2.00M CH3 COOH

ALTERNATIVE TWO APPROACH for calculating # of mols of each buffer component from
[PA]/[PD] ratio:

1.0965/1 = [CH3 COO-]present/[CH3 COOH]present

[CH3 COOH]present = (Total buffer concn) - [CH3 COO-]present

ˆ [CH3 COOH]present = (0.10 M) - [CH3 COO-]present

1.0965/1 = [CH3 COO-]present/(0.10 M) - [CH3 COO-]present

0.10965 M - (1.0965)[CH3 COO-]present = [CH3 COO-]present

0.10965 M = [CH3 COO-]present + (1.0965)[CH3 COO-]present

0.10965 M = {[CH3 COO-]present}(1 + 1.0965)

0.10965 M/2.0965 = [CH3 COO-]present = 0.0523 M

ˆ [CH3 COOH]present =(Total buffer concn - 0.0523M) = (0.10 M - 0.0523 M) = 0.0477 M

Thus, # moles in 500 ml 0.1M acetate buffer pH 4.8:

[CH3 COO-] = (0.0523 mol/L)(0.5L) = 0.02615 moles in 500 ml

[CH3 COOH] = (0.0477 mol/L)(0.5L) = 0.02385 moles in 500 ml

The Rest of the Calculation is as given above after Alternative One approach

2. A buffer solution is 0.10 M in acetic acid, and 0.05M in sodium acetate. Calculate the pH of
this buffer solution, if the pKa of acetic acid is 4.76.
(Ans. = 4.46)

The relevant weak acid dissociation is: CH3 COOH + H2 O » CH3 COO- + H3 +O

[Note: From the wording of this question, you have been provided with the proton acceptor and proton
donor concns that exist in the buffer.]

SOLUTION:
Concn of CH3COO- = 0.05M
Concn of CH3COOH = 0.10M
pH = pKa + log{[CH3COO-]/[CH3COOH]}
pH = 4.76 + log{0.05/0.1}
pH = 4.76 + (-0.301)
pH = 4.459 = 4.46

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 3 You have been provided with 200 ml of a 0.10 M aqueous solution of NaH2 PO4 , calculate the
number of grams of Na2 HPO4 which you must add to this NaH2 PO4 solution in order to create
a buffer with a pH of 6.70. [Assume that (i) the addition of solid Na2 HPO4 does not alter the
volume; (ii) For phosphoric acid: pKa1 = 2.15; pKa2 = 6.82; pKa3 = 12.38; and Formula
weight of NaH2 PO4 = 141.96]
(Ans. = 2.15 g Na2 HPO4 )

SOLUTION:

H3PO4 W H+ + H 2PO4- W H+ + HPO42- W H+ + PO43-

pKa= 2.15 6.82 12.38

pH = pKa + Log{[HPO4 2-]/[ H2 PO4 -]}

6.7 = 6.82 + Log{[HPO4 2-]/[ H2 PO4 -]}
10-0.12 = [HPO4 2-]/[ H2 PO4 -]
0.7586/1 = [HPO4 2-]/[ H2 PO4 -]
0.7586/1 = [HPO4 2-]/0.10M
(0.10M)(0.7586) = [HPO4 2-]

In this buffer: There is 0.07586 mol [HPO4 2-]/L
Molar mass of Na2 HPO4 = 141.96g/mol
So, gm required = (141.96 g/mol)(0.015172mol)
= (0.07586 mol HPO4 2-]/L)(0.2)L
= 0.015172 mol HPO4 2-/200 ml
= 2.1538g Na2 HPO4
= 2.15 g Na2 HPO4 /200ml buffer

4. Calculate the concentration of ammonia (NH3 ) (in moles per litre) in an aqueous ammonium
hydroxide (NH4 OH), given that 0.1043 g of NH4 Cl dissolved in 30 ml distilled water required
22.0 ml of the NH4 OH solution to create a buffer of pH 8.7.
(Ans. = 0.025 mol/L)

Solution:
The relevant weak acid dissociation = NH4 + » NH3 + H+

0.1043 g NH4 CL = 0.1043g/(53.5g/mol) = 0.00195 mol NH4 Cl

8.7 - 9.25 = log([NH4 OH]/[NH4 CL)]

10-0.55 = [NH4 OH]/[NH4 CL]

0.282/1 = [NH4 OH]/[NH4 CL]

So, mol fractions: NH4 CL (in the buffer) = 1/1+0.282 = 0.78

NH4 OH (in the buffer) = 1 – 0.78 = 0.22

Thus, 0.78 = 0.00195 mol of NH4 CL

Therefore: 0.22 = (0.00195 mol/0.78) x 0.22 = 0.00055 mol NH4 OH

This means that: 0.00055 mol NH4 OH = 0.00055 mol NH3 in 22 mL

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Therefore: # mol NH3 in 1 L = (0.00055 mol NH3 /22 mL) x (1000mol/L) = 0.025 mol/L

Q7 Calculate the equilibrium constant for the following reaction at pH 7.0 and 25o C, given that
the ∆G’o = -20.9 kJ.mol-1 and the gas constant R = 8.315 J.mol-1 .K-1 :

Glucose-1-phosphate + H2 O ÷ glucose + H2 PO4 -

[Ans: K eq = 4604]

Solution:
∆G’o = -RTlnKeq
-1
-20.9 kJ.mol = [-(8.315x10-3 kJ.mol-1 .K-1 )(25+273)K]lnK eq
-20.9 kJ.mol-1 = -(8.315x10-3 kJ.mol-1 .K-1 )(298)K]lnK eq

lnKeq = (-20.9 kJ.mol-1 )/[(-8.315x10-3 kJ.mol-1 .K-1 )(298K)] = 8.4347

Therefore, Keq = e8.4347 = 4604

Q8 You are provided with 100 ml of 0.2 M glutamate solution at pH = pI.

(a) Sketch with clearly labeled volume and pH axes, the curve resulting from the complete
titration of this solution with 0.1 M aqueous NaOH. Indicate on your sketch the volumes of
NaOH solution added to reach the relevant pKa value(s). Show clearly how you arrived at the
volume(s). [pKa values are 2.19, 4.25 (R group) and 9.67].
[Ans: 100 ml 0.1M NaOH to reach half titration (i.e., pKa) of the γ-COOH proton
300 ml to reach half titration (i.e., pKa of the α -N+H3 proton]

(b) Calculate, being sure to show how you arrived at your answer, the volume of 0.1 M NaOH
needed to reach pH = 5.0 starting from pH = pI.
(Ans: 170 ml 0.1M NaOH).

(c) Draw the structure of the form of glutamate predominating in solution at pH 5.0.

SOLUTION:
_ _ _
COOH pKa COO pKa
pKa COO COO
9.67
H3+N CH 2.19 H3+N CH 4.25 H3+N CH H2N CH
CH2
H+ + CH2
H+ + CH2 H+ + CH2
CH2 CH2 CH2 CH2
COOH COOH COO _ COO_
+
Glu o
Glu Glu- Glu2-
Zwitterion

(a) Given: 100 ml 0.2M glutamate @ pH = pI (So, Structure = Gluo = Zwitterion)

pI = (2.19 + 4.25)/2 = 3.22

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100 ml 0.2M glutamate = (0.2 mol/L)(0.1L) = 0.02 mol glutamate

100 ml 0.2M glutamate @ pI = 0.02 mol Gluo

So, contains Total of 0.04 mol protons from the following sources:

0.02M of α-N+H3 protons

And, 0.02M of γ-COOH protons

For complete titration, the 0.04 mol H+ requires 0.04 mol OH-

0.01M NaOH is provided:

1L contains 0.1 mol OH- So, 0.04 mol is in {(1L)/(0.1mol)}(0.04mol)}
Therefore, 0.04 mol is in 0.4 L.

0.02 mol γ-COOH @ pI requires 0.2L (= 200ml) 0.1M NaOH for titration completely
0.02mol of α-N+H3 @ pI requires 0.2L (= 200ml) 0.1M NaOH for titration completely

To reach pH = pKa = 4.25 (midpoint of Gluo W Glu- titration)

{(Requires 200ml)/2} = 100 ml 0.1M NaOH

To reach pH = pKa = 9.67 (midpoint of Glu- W Glu2- titration)

{(Requires 200ml + (200ml/2)} = 300 ml 0.1M NaOH

(b) Vol 0.1M NaOH required to reach pH = 5.0 from pI

At pH = 5.0 there will be more Glu- than Gluo

pH = pKa + Log{[Glu-]/[ Gluo ]}

5.0 = 4.25 + Log{[Glu-]/[ Gluo ]}
10(5.0-4.25) = 100.75 = 5.62/1 = [Glu-]/[ Gluo ]

Fraction as Glu-] = 5.62/6.62 = 0.85

[Total glutamate = 0.02 mol

So, moles Glu- @ pH5 = (0.02 mol)(0.85) = 0.017 mol

Fraction as Gluo ] = 1/6.62 = 0.15

So, moles Gluo @ pH5 = (0.02 mol)(0.15) = 0.003 mol

Therefore, NaOH required to form 0.017 mol Glu- @ pH 5.0 [from 0.02 mol initial Gluo (pH 3.22)]

= 0.017 mol NaOH

Given 0.1M NaOH

0.017 mol NaOH is contained in (1L/0.1M)(0.017mol/1) = 0.17L 0.1M NaOH

= 170 ml 0.1M NaOH

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 (c) Structure of glutamate predominating in solution @ pH 5.0

_
COO
H3+N CH
CH2
CH2
COO_

Glu-

Q11. Given two buffer solutions, A and B:

Buffer solution A contains 0.05 M borate and 0.1M boric acid

Buffer solution B contains 0.01 M borate and 0.02M boric acid

(a) Suggest with reasons which of the two buffers has a higher buffer capacity
(b) Calculate the pH of each buffer, given that the pKa is 9.25 for boric acid.

SOLUTION:
Relevant weak acid dissociation: H3 BO3 » H+ + H2 BO3 -

(a) Concns of weak acid and the conj. base are 5-fold higher in A than in B
But in each of buffer A and Buffer B, the ratio [H2 BO3 -]/[H3 BO3 ] = 0.5

Therefore, buffer A has a higher buffer capacity than buffer B

Reason: being more conc, buffer A will be able to stand a larger amount of acid or base added to it without
appreciable change in the pH.

(b) For buffer A

pH = pKa + Log{[H2 BO3 -]/[H3 BO3 ]}

pH = 9.25 + log(0.05/0.01) = 9.25 – log(0.5) = 9.25 – 0.3010 = 8.95

For buffer B:
Same value of 0.5 for [H2 BO3 -]/[H3 BO3 ] Therefore same pH = 8.95.