INVESTIGATION 16.

THE PREPARATION AND

TESTING OF AN EFFECTIVE
BUFFER: HOW DO
COMPONENTS INFLUENCE A
BUFFER’S PH AND CAPACITY?
ABSTRACT: THE PURPOSE OF THIS LAB IS TO PREPARE AND TEST A BUFFER SOLUTION
WITH A GIVEN PH, THEN DETERMINE IF THE BUFFER SOLUTION WE PREPARED IS
ADEQUATE AND RELIABLE ENOUGH TO BE USED FOR AN IMPORTANT BIOCHEMICAL
PROJECT, OR IF A SOLUTION MADE BY ANOTHER GROUP IS NEEDED.

EDWARD MOON – PRELAB, DATA COLLECTION AND COMPUTATION

DANA CHOI – PROCEDURE, POSTLAB
ANTHONY HOWER – PRELAB, ABSTRACT, PROCEDURE, CONCLUSION
Table of Contents
Prelab guiding questions .................................................................................................. 3

Procedure .......................................................................................................................... 5

Data and calculations ....................................................................................................... 7

Post-lab assessment ......................................................................................................... 9

Conclusion ....................................................................................................................... 11
Prelab guiding questions
1. How does a buffer solution resist a change in pH?
A buffer solution resists a change in pH because it is in a state where there are
both acid and its conjugate base molecules (or base and its conjugate acid), so
that when acid or base is added, it will be neutralized. This can only be done if a
weak acid or base is involved. If the acid is weak and the base is strong, a basic
buffer will form, and if the base is weak and the acid is strong, an acidic buffer
will form

2. Why would HCl and NaOH be a poor choice for an acid-base pair to make a
buffer?
HCl and NaOH would be a poor acid-base pair choice for a buffer because they
are both strong, and equal amount of them will cancel each other out, and
their salt has no buffering ability. A weak acid or base needs to be added in
order to form a buffer.

3. Go to the animation at:

http://introchem.chem.okstate.edu/DCICLA/pHbuffer20.html
Create a “buffer solution” using equal volumes (100 mL each) of 0.1 M nitric
acid and 0.1 M sodium nitrate. Insert the probes and record the pH in the table
below. Remove the probes. Then go to Part II of the solution and add first
0.001 moles of HCL and then the same amount of NaOH. Record the pH in each
case. Remove the probes, return to Part I, and try the next combinations.
Complete the first three lines of the table below.

Remake the “buffers” using the same volumes of 1.0 M components (increased
molarity by 10X). Add the same 0.001 moles of HCl and NaOH to the “strong”
buffers. Complete the middle of the table.

Finally, repeat the process adding 0.011 moles (additional 10X more) of strong
acid and base. Complete the last three lines of the table.

pH with 0.001 mol pH with 0.001 mol

Acid-Base Pair pH of “Buffer”
HCl added NaOH added

HNO3 and NaNO3 1.30 1.25 1.34

HC2H3O2 and
4.74 4.65 4.83
NaC2H3O2

NH4Cl and NH3 9.25 9.16 9.34

Increase Molarity
10X

HNO3 and NaNO3 0.30 0.29 0.30

HC2H3O2 and
4.74 4.73 4.75
NaC2H3O2

NH4Cl and NH3 9.25 9.24 9.26

pH with 0.011 mol pH with 0.011 mol

HCl added NaOH added

HNO3 and NaNO3 0.30 0.25 0.35

HC2H3O2 and
4.74 4.64 4.84
NaC2H3O2

NH4Cl and NH3 9.25 9.15 9.35

a. Write a general chemical equation to represent the equilibrium that exists in

an aqueous system of the weak acid, HA as it ionizes in water. Assume the
weak acid to be HC2H3O2 (0.10 M), with a Ka value of 1.8 x 10-5. What is the
pH? What does the addition of NaC2H3O2 do to the equilibrium you just
represented? Use these chemical principles to explain why the first pH
recorded from the simulation is so much larger than your calculated value
for pure 0.10 M acetic acid.
A general chemical equation for an aqueous system of the weak acid is: HA
(aq) + H2O (l) ↔ H3O+ (aq) + A- (aq), and when the weak acid is HC2H3O2, the
equilibrium equation would look like this: HC2H3O2 (aq) + H2O (l) ↔ H3O+ (aq)
+ C2H3O2- (aq). Based on this equation, we can set up an ICE table to calculate
the concentration of H3O+ in the solution, and the calculated pH would equal
to 2.89. The addition of NaC2H3O2 would increase the concentration of C2H3O2-
in the equilibrium reaction above, shifting the reaction to the left, which
would eventually cause a decrease in pH. Then, the solution would become
basic as more NaC2H3O2 is added.

b. How do the pKa values for acetic acid and ammonium ion (Ka = 5.6 x 10-10)
compare to the pH values for the first two buffers in the simulation? Explain
this phenomenon.
The lower the pKa value, the strong the acid. The pKa value for acetic acid and
ammonium ion is 9.25, while the pH values for the first two buffers were 1.30
 and 4.74. This indicates that acetic acid and ammonium ion are weak acids,
and that the first two buffers are more acidic. The reason is because nitric acid
(HNO3) in the first buffer is strong acid, and NaC2H3O2 is a weak base.

c. Why don’t the pH values change when the component concentrations are
increased?
Buffering capacity refers to the ability of the buffer to resist changes in pH
when acid or base is added. When the “strong buffers” with higher
concentrations are used, its buffering capacity increases, and therefore, pH
does not change as easily.

4. Write a general net ionic equation to show how a buffer containing an acid,
HA, and the salt of its conjugate base, NaA, would respond to the addition of
each of the following.
a. The strong acid, HCl
A-(aq) + H3O+(aq) --> H2O(l) + HA(aq)

b. The strong base, NaOH

OH-(aq) + HA(aq) --> H2O(l) + A-(aq)

5. Why are the pH changes so noticeable with the last two additions of strong
acid and base in the simulation?
Because greater concentrations of HCl and NaOH are added to the solutions.
Buffers are less resistant to changes in pH when high concentrations of strong
acid or base are added.

6. Which of the two concentration combinations would be most effective for

any buffer you might design?
1.0 M acid / base and salt. Buffer made of those with greater concentration
would be more resistant to changes in pH.

Procedure
Before starting the lab, set up the apparatus. Put a magnetic stir-plate next to
a vertical pole-stand, and the drop counter and buret suspended with space for a
beaker below, and the pH probe low enough to reach the bottom of the space for the
beaker. Once all the wires for the probes are in place and hooked up to the computer
with Logger Pro open, calibrate the probes. First, fill the buret with a liquid such as
water and adjust the speed valve to the desired speed. Do not touch the speed valve
again. In Logger Pro, open the drop counter calibration and click start. Open the top
valve and record the exact volume of liquid dropped and enter it into the computer to
calibrate the mL per drop, so that data collection will be accurate. If needed, the pH
probe can be calibrated by submerging it in a known low-pH buffer solution, inputting
into the computer what the pH is, then rinsing it and doing the same with a high pH
solution, in order to make the pH readings more accurate.
Hypothesis: If we combine sodium monohydrogen citrate (Na2HC6H5O7) and sodium
citrate (Na3C6H5O7), then our buffer will neutralize HCl and NaOH.

The goal of this lab is to make a buffer solution that contains equal
concentrations of sodium bicitrate (Na2HC6H5O7) and sodium citrate (Na3C6H5O7), with
a net-ionic equation of: Na2HC6H5O7 (aq) + H2O (l) ↔ H3O+ (aq) + Na3C6H5O7 (aq). But
since there is only citric acid (H3C6H5O7) and not sodium monohydrogen citrate
(Na2HC6H5O7) and sodium citrate (Na3C6H5O7) to start with, three steps of chemical
reactions should be completed.

Step 1: H3C6H5O7 + NaOH ↔ NaH2C6H5O7 + H2O

Start with 100mL of 0.1M citric acid to obtain sodium dihydrogen citrate
(NaH2C6H5O7). Add 50mL of 0.2 M NaOH and completely react citric acid and obtain
150mL of sodium dihydrogen citrate (NaH2C6H5O7).

Step 2: NaH2C6H5O7 + NaOH ↔ Na2HC6H5O7 + H2O

Then, to 150mL of sodium dihydrogen citrate (NaH2C6H5O7), add 50mL of

NaOH and completely react sodium dihydrogen citrate and obtain 200 mL of sodium
bicitrate (Na2HC6H5O7).

Step 3: Na2HC6H5O7 + NaOH ↔ Na3C6H5O7 + H2O

Finally, to 200mL of sodium bicitrate (Na2HC6H5O7), add 25mL of NaOH so that

half of sodium bicitrate (Na2HC6H5O7) will react to form sodium citrate (Na3C6H5O7).

The buffer solution obtained from these steps will contain equal concentrations of
sodium bicitrate (Na2HC6H5O7) and sodium citrate (Na3C6H5O7).

Now, equally spit the buffer solution into two beakers. Put one of the beakers over the
stir plate, with the drop counter and buret over it, and pH probe submerged in the
buffer solution. If the solution is not high enough to reach the probe, add some
distilled water, as adding water does not affect the number of moles in the solution,
which is what matters for this lab. To do the base titration first, fill the buret with
about 20 mL of 0.2M NaOH. Turn on the magnetic stir. Once data recording is turned
on on the computer recording the data, open the top valve on the buret to start
titrating in the base. Once the buret is empty or sufficient data is obtained, close the
valve and save the data. Reset the setup by rinsing the pH probe and buret, then fill
the buret with about 20mL 0.2M HCl and put the other buffer beaker under the
apparatus to do the acid titration. Repeat the same process as the base titration and
then clean up the lab.
 pH pH

5
6

4.6
4.8
5.2
5.4
5.6
5.8
6.2
10
11
12
13

5
6
7
8
9
10.5
11.5
12.5

5.5
6.5
7.5
8.5
9.5
0
0.035714286
Data and calculations

0.785714286
0.785714286
1.571428571
1.535714286
2.357142857
2.285714286
3.142857143
3.035714286
3.928571429 3.785714286
4.714285714 4.535714286
5.5 5.285714286
6.285714286 6.035714286
7.071428571 6.785714286
7.857142857

pH
pH

7.535714286
8.642857143 8.285714286
9.428571429 9.035714286
10.21428571 9.785714286
11 10.53571429
11.78571429 11.28571429
12.57142857 12.03571429
13.35714286 12.78571429
14.14285714 13.53571429
Amount of 0.2M HCl Added

Amount of Acid added vs.

14.92857143
Amount of Base added vs.

Amount of 0.2M NaOH Added

14.28571429
15.71428571
1. Complete neatly labeled graphs for the data tables you’ve prepared.

15.03571429
16.5 15.78571429
17.28571429 16.53571429

Buffer: Sodium monohydrogen citrate (Na2HC6H5O7) & Sodium citrate (Na3C6H5O7).

2. Calculate the anticipated pH for the buffer you were assigned. Include neat
calculations in your report.

We expected our buffer to contain the same concentration of HA (acid) and

A- (conjugate base) – the equal concentration of these two indicates that the buffer is
capable of neutralizing both acids and bases.

Our buffer contained sodium monohydrogen citrate (Na2HC6H5O7) and sodium citrate
(Na3C6H5O7), and the general weak acid equation would look like this:

Na2HC6H5O7 (aq) + H2O (l) ↔ H3O+ (aq) + Na3C6H5O7 (aq).

[𝐻+][𝐴−]
Thus, 𝐾𝑎 = [𝐻𝐴]
.

We expect [HA] to be equal to [A-]; those two will cancel out, and Ka = [H3O+].

Therefore, pKa = pH. We are given Ka value, which is 4.1 x 10-7.

So pH = -log [4.1 x 10-7] = 6.39.

3. Determine the volume of acid and base your buffer was able to neutralize before
allowing a pH change of more than one unit. Use the volume with the HCl / NaOH
molarities to determine the number of moles of acid and base your buffer was able
to neutralize.

Reactions:

Adding Base: Na2HC6H5O7 (aq) + OH- (aq) ↔ Na3C6H5O7 (aq) + H2O (l)

Adding Acid: Na3C6H5O7 (aq) + H3O+ (aq) ↔ Na3HC6H5O7 (aq) + H2O (l)

Our buffer was able to neutralize 5.5 mL of 0.2M NaOH and about 18 mL of 0.2M HCl.
0.2 𝑚𝑜𝑙𝑒𝑠 𝑁𝑎𝑂𝐻 1𝐿
NaOH: ∗ 1000 𝑚𝐿 ∗ 5.5 𝑚𝐿 = 1.1 x 10-3 moles NaOH.
1𝐿

0.2 𝑚𝑜𝑙𝑒𝑠 𝐻𝐶𝑙 1𝐿

HCl: ∗ 1000 𝑚𝐿 ∗ 18 𝑚𝐿 = 3.6 x 10-3 moles HCl.
1𝐿
4. Consider 50.0 mL of the buffer you prepared and calculate the ratio of (moles of
added acid neutralized) to (moles of base component in the buffer). Do the same
calculation for (moles of added base neutralized) to (moles of acid component in the
buffer). State both ratios in the form of x:1. Report the value for x to two significant
figures.

Acid:

When Acid Added: Na3C6H5O7 (aq) + H3O+ (aq) ↔ Na3HC6H5O7 (aq) + H2O (l)

Molarity of Na3C6H5O7 = 0.10 M

0.10 𝑚𝑜𝑙𝑒𝑠 1𝐿
Moles of Na3C6H5O7 = ∗ 1000 𝑚𝐿 ∗ 50𝑚𝐿 = 5.0 x 10-3.
1𝐿

Ratio of (moles of added acid) to (moles of base component in the buffer) =

0.0036: 0.0050 = 0.72: 1.

Base:

When Base Added: Na2HC6H5O7 (aq) + OH- (aq) ↔ Na3C6H5O7 (aq) + H2O (l)

Molarity of Na3HC6H5O7 = 0.10 M

0.10 𝑚𝑜𝑙𝑒𝑠 1𝐿
Moles of Na3HC6H5O7 = ∗ 1000 𝑚𝐿 ∗ 50𝑚𝐿 = 5.0 x 10-3.
1𝐿

Ratio of (moles of added base) to (moles of acid component in the buffer) =

0.0011: 0.0050 = 0.22: 1.

Post-lab assessment
1. Suppose, during preparation, an additional 10 mL of distilled water was
added to your buffer by mistake.
a. What effect would this have on your buffer’s pH? Explain.
HA (aq) + H2O (l) ↔ H3O+ (aq) + A- (aq).
The addition of water will shift the equilibrium to the right and increase
the concentration of the conjugate base, which will increase the pH.

b. Would this affect your buffer’s capacity? Explain.

The buffer capacity to neutralize base will increase but the capacity to
neutralize acid will stay the same.
2. Given a solution of hydrocyanic acid (HCN), what additional reagent or
reagents is/are needed to prepare a buffer from the hydrocyanic acid
solution?
HCN (aq) + NaOH (aq) ↔ H2O (l) + NaCN (aq)
a. Explain how this buffer solution resists a change in pH when moderate
amounts of strong acid are added. Use a chemical equation in your
explanation.
The conjugate base in the buffer solution, which is CN -, neutralizes any
added acid and increases the concentration of HCN.
CN- + H+ → HCN

b. Explain how this buffer solution resists a change in pH when moderate

amounts of strong base are added. Again, provide an equation with
your explanation.
The acid in the buffer solution, which is HCN, neutralizes any added
base and increases the concentration of CN-.
HCN + OH- → CN- + H2O

3. A buffer solution contains 0.20 moles of methanoic acid, HCOOH, and 0.30
moles of sodium methanoate, NaCOOH, in 1.00 L of the buffer. The acid
ionization constant, Ka, of methanoic acid is 1.8 × 10–4.
a. Calculate the pH of this solution.
According to the Henderson-Hasselbalch equation,
[𝐴−]
𝑝𝐻 = 𝑝𝐾𝑎 + 𝑙𝑜𝑔([[𝐻𝐴])

pKa=-log(1.8 × 10–4)=3.74, [A-]=0.3M, [HA]=0.2M

 pH=3.74 + log(0.3/0.2)=3.92

b. Compare the capacity of this buffer to neutralize added acid to its

capacity to neutralize added base. Explain your answer completely.
Since there are more amount of conjugate base than acid in this buffer
solution, the buffer capacity to neutralize added acid is higher than that
of added base.
 c. If 0.10 moles of HCl gas solution were bubbled through a liter of the
buffer, what would happen to the pH? How would this addition affect
the buffer’s capacity to neutralize added acid and base in the future?
Answer the question fully, including equations and calculations where
necessary.
When HCl is added to the buffer: COOH- + H3O+ → HCOOH + H2O
Here, when 0.10 moles of HCl is added, COOH- decreases by 0.10 moles
and HCOOH increases by 0.10 mole. Therefore, the amount of HCOOH
and COOH- will both be 0.30 moles. Thus, [A-]:[HA]=1:1 and the buffer
capacity to neutralize acid will equal the buffer capacity to neutralize
base. Also, since the concentrations of HCOOH and COOH- will be the
same, pH will equal to pKa. Therefore, the new pH will decrease to 3.74.
(pH=pKa=-log(1.8 × 10–4)=3.74)

Conclusion
The purpose of our lab was to create and test a buffer solution given a certain pH that
would be reliable enough to use in an important biochemical project. To be deemed
reliable enough, a 50 mL solution of our buffer should be able to withstand the
addition of 20mL 0.2M NaOH or HCl, while maintaining a relatively constant pH, with a
difference within one pH unit of the initial value. The pH that we were designated was
6.4, and thus we chose to make our buffer using monohydrogen citrate, which has a K a
of 4.1 X 10-7, translating to a pH of 6.39, essentially a perfect match for our given
solution value. However, we did not have this chemical lying around and thus had to
make the chemical from citric acid and Sodium Hydroxide, as well as its conjugate in
order to make our buffer solution. After testing our buffer solution, while the buffer
remained within 1 pH unit of the initial for up to 15 mL of HCl, the pH only remained
stable for about 6.5 mL of NaOH. Therefore, as our buffer was not able to withstand
the addition of acid or base well, we determined that another group’s buffer solution
should be used as the one we developed is not reliable enough.