Acid-Base Chemistry:

The Analytical Preparation of a Buffer and a Primer of Essential

Laboratory Skills

Purpose

During this two-week experiment, you will calibrate volumetric glassware and prepare buffers with a target pH.
In addition, you will measure the buffering capacity of several solutions.

Learning Objectives
Calibrate laboratory equipment with a focus on volume-measuring equipment.

Create a calibration curve to correct volume measurements.

Make a buffer solution.

Use a micropipette, a volumetric flask, a balance, and a pH probe.

Analyze buffer capacity and to understand its significance.

Write one section of the laboratory report. See Sakai and the Experiment 4 Addendum for more infor-
mation.

Reading Assignment

Required Reading: Read about the following topics from your analytical chemistry text. The chapter numbers
and sections listed are from Quantitative Chemical Analysis, 9th ed. by D.C. Harris.

• Calibration Topics (2-2, 2-3, 4-8)

• Buffer and Buffer Capacity Topics (6-6, 8-2–8-5, 9-2)
• pH Probe Use (14-5)
• See Canvas for any additional supplements and additions to this experiment!

Laboratory Manual Prepared by Catalyst Education, LLC for University of North Carolina University at Chapel Hill BUS.1
Department of Chemistry.
 Part II: Preparation and Measuring the Buffer Capacity of Target Buffers

Background and Theory

Preparation of Buffered Solutions

In the second part of this lab, you will prepare two buffers at a pH of 4.50. One buffer will be prepared using a
theoretical approach, and the second will be prepared using a more practical approach. You will then compare
and contrast the buffer capacity of these two buffers with the control experiments you did in week 1.

When preparing to make a buffer solution, it is important to choose a buffer whose p𝐾a is as close to the desired
pH as possible.1 You typically will not find a buffer whose p𝐾a exactly equals the desired pH, so you will need to
choose a buffer whose p𝐾a is as close to the desired pH as possible, within ± 1 pH unit.

Example BUS.1

You wish to prepare a buffer of pH 6.00. Ideally it would be nice to find a buffer whose p𝐾a is
6.00, but it is acceptable to be within ± 1 pH unit (e.g., 5.00 to 7.00 is an acceptable range, but as
close as possible to 6.00).

Theoretical Approach to Making Buffers

The theoretical approach involves selecting two reagents, based on your assigned pH, and using the
Henderson-Hasselbalch equation to calculate the amount of each reagent needed. You then add those
reagents together into a volumetric flask of the desired volume followed by dilute to the line. For this
experiment you will need to select a weak acid —weak base conjugate pair.

Example BUS.2

Weak Acid (HA) and Its Conjugate Base (A – )

A solution is prepared by adding 2.5 g of formic acid (HCO2 H; MW = 46.025 g/mol) and 3.5 g of
sodium formate (HCO2 Na; MW = 68.005 g/mol) to a 1.00-L volumetric flask and diluting with
distilled water to the mark.

A buffer is formed because a weak acid (formic acid) is added to its conjugate base (for-
mate). The pH can be calculated using the Henderson-Hasselbalch equation:

0.0515 mol/L
pH = 3.744 + 𝑙𝑜𝑔 = 3.72
0.0543 mol/L

Laboratory Manual Prepared by Catalyst Education, LLC for University of North Carolina University at Chapel Hill BUS.2
Department of Chemistry.
 Part II: Preparation and Measuring the Buffer Capacity of Target Buffers

This week, in week 2 of the experiment, you will complete the math necessary to determine how much of a weak
acid and its conjugate base you will need to add to make 100 mL of a buffer at pH 4.5.

Practical Approach to Making Buffers

The practical approach involves choosing one of the reagents you selected above (either the weak acid
or weak base) and using either a strong acid or a strong base to form the conjugate. There are two ways
to do this. If you select the weak acid, you will need to partially neutralize it with a strong base to form the
conjugate base of the weak acid. If you select the weak base, you will need to partially neutralize it with a strong
acid to form the conjugate acid of the weak base. You do not need to calculate the amount of strong acid or strong
base to add; you will simply monitor the pH as you add the strong acid or strong base. Once you have selected
your reagents, follow the protocol below in order to make your buffer using the practical approach (as outlined
in the Harris textbook on p. 202 and also linked on Sakai). In both of the methods above, the general procedure
is the same:

1. Weigh out the desired number of moles of your weak acid OR weak base and dilute in a beaker con-
taining 80 mL of deionized water.
2. Place a calibrated pH electrode in the solution to monitor the pH.
3. Add either strong acid or strong base into the beaker until your desired pH (4.50) is met.
4. Transfer the solution to a 100.00-mL volumetric flask; wash the beaker with deionized water and add
the washings to the flask as well. Dilute to the line with water and mix.

Partial Neutralization of a Weak Acid with a Strong Base

If you started with a weak acid in step 1 above, but not its conjugate base, you can use a strong base to partially
neutralize the weak acid to generate the conjugate species.

HA + OH− −−−→ A− + H2 O

Weak Acid + Strong Base −−−→ Conjugate Base + Water

Laboratory Manual Prepared by Catalyst Education, LLC for University of North Carolina University at Chapel Hill BUS.3
Department of Chemistry.
 Part II: Preparation and Measuring the Buffer Capacity of Target Buffers

Example. A solution is prepared by adding 12.0 mL of 1.0 M NaOH to a 1.0-L solution containing 0.05 moles of
formic acid (HCO2 H; p𝐾a = 3.744).

HCO2 H + OH – −−−→ HCO2 – + H2 O

Initial Moles 0.05 mol 0 mol 0 mol

Added Moles 0 (0.012 L)(1.0 M) 0 mol

(0.05-0.012) mol 0 (reacted) 0.012 mol

The final pH of this solution would be:

0.012 mol
pH = 3.744 + 𝑙𝑜𝑔 = 3.24
0.038 mol

Observations regarding buffer solutions:

a The amount of added strong base is equal to the amount of weak acid you lose and the same amount of
conjugate base you produce.
b The moles of weak acid are greater than the moles of added strong base, so you know you are producing
a buffer via partial neutralization.
c The pH of the buffer solution depends on the ratio of conjugate base to weak acid and thus you can
simply use moles in the Henderson-Hasselbalch, you do not need to convert back to concentrations.

Partial Neutralization of a Weak Base with a Strong Acid

If you started with a weak base in step 1 above, but not its conjugate acid, you can use a strong acid to partially
neutralize the weak base to generate the conjugate species.

A− + H+ −−−→ HA

Weak Base + Strong Acid −−−→ Conjugate Acid

Example. A solution is prepared by adding 12.0 mL of 1.0 M HCl to a 1.0-L solution containing 0.05 moles of
formate (HCO2 – ; p𝐾a (HCO2 H) = 3.744).

HCO2 – + H+ −−−→ HCO2 H

Initial Moles 0.05 mol 0 mol 0 mol

Added Moles 0 (0.012 L)(1.0 M) 0 mol

(0.05-0.012) mol 0 (reacted) 0.012 mol

Laboratory Manual Prepared by Catalyst Education, LLC for University of North Carolina University at Chapel Hill BUS.4
Department of Chemistry.
 Part II: Preparation and Measuring the Buffer Capacity of Target Buffers

The final pH of this solution would be:

0.038 mol
pH = 3.744 + 𝑙𝑜𝑔 = 4.24
0.012 mol

The same three observations listed above apply to this weak base/strong acid example.

Laboratory Manual Prepared by Catalyst Education, LLC for University of North Carolina University at Chapel Hill BUS.5
Department of Chemistry.
 Part II: Preparation and Measuring the Buffer Capacity of Target Buffers

Experimental

Part 2.1: Selection and Calculation of the Reagents to Prepare Target Buffer

YOU AND YOUR PARTNER CAN WORK ON THIS PART TOGETHER BEFORE COMING TO LAB! Please
read the posted “How to prepare a buffer” section from the Harris textbook (posted on Canvas) prior to starting.

You and your lab partner will be preparing a buffer this week at pH 4.5. You and your lab partner will work
together to calculate the amount of each reagent you will need to prepare these buffers. Write out all your work
in your laboratory notebook.

1. Select two reagents from the list below that you will use in lab to prepare the theoretical buffer solution.
Discuss why these reagents were selected and why they were the best choice. You may need to reference
additional information to make your choice.

Table BUS.1: Possible reagents for the theoretical buffer solution.

Reagent Formula Molecular Weight for

Solids or Concentration for
Liquids

Acetic acid CH3 CO2 H 0.50 M

Sodium acetate, trihydrate CH3 CO2 Na ⋅ 3 H2 O 136.08 g/mol

Sodium citrate, dibasic C6 H6 Na2 O7 ⋅ 1.5 H2 O 263.11 g/mol

Sodium acetate, tribasic C6 H5 Na3 O7 ⋅ 2 H2 O 294.1 g/mol

Sodium phosphate, monobasic NaH2 PO4 ⋅ H2 O 137.99 g/mol

Sodium phosphate, dibasic Na2 HPO4 141.96 g/mol

Sodium phosphate, tribasic Na3 PO4 ⋅ 12 H2 O 380.12 g/mol

Sodium hydrogen sulfate, NaHSO4 ⋅ H2 O 138.07 g/mol

monohydrate

Sodium sulfate Na2 SO4 142.04 g/mol

Sodium carbonate, monohy- Na2 CO3 ⋅ H2 O 124.0 g/mol

drate

Sodium bicarbonate NaHCO3 84.01 g/mol

Laboratory Manual Prepared by Catalyst Education, LLC for University of North Carolina University at Chapel Hill BUS.6
Department of Chemistry.
 Part II: Preparation and Measuring the Buffer Capacity of Target Buffers

2. Which reagents (one of the reagents you selected in Question 1 and one from the table below) will you use
to prepare the practical buffer solution? Discuss why these reagents were selected and why they were the
best choice.

Table BUS.2: Possible reagents for the practical buffer solution.

Reagent Formula Molecular Weight for

Solids or Concentration for
Liquids

Sodium hydroxide NaOH 0.5 M

Hydrochloric acid HCl 0.5 M

3. Theoretical Buffer Solution: Using the reagents you selected in question 1, prepare a 0.050 M buffered
solution using the reagents you selected in question 1. You will prepare 100 mL of buffered solution with
pH = 4.5. The formal concentration of this buffer should be 0.050 F (i.e., the total concentration of the
weak acid and its conjugate base is 0.050 M).

• Determine the mass or volume for both the reagents that you will need to prepare the above buffer.
• Record your calculations, clearly showing all your work with proper labels (e.g., mass of reagent A, etc.).
• Based on the pre-lab reading, write a short procedure in your notebook. Have the TA check your pro-
cedure.

4. Practical Buffer Solution: Using the reagents you selected for question 2, describe how you would prepare
100 mL of a 0.050 M buffer to a pH of 4.5 based on the method described on p. 202 of your Harris textbook:
“Preparing a Buffer in Real Life!” This section is included in the excerpt on the Canvas website in the Exper-
iment 4 Module. Outline a procedure in bullet form and make sure to include what amounts of the weak acid
or weak base you would need to dissolve and which strong acid or base you will use based on your calculations.
Have the TA check your procedure.

Laboratory Manual Prepared by Catalyst Education, LLC for University of North Carolina University at Chapel Hill BUS.7
Department of Chemistry.
 Part II: Preparation and Measuring the Buffer Capacity of Target Buffers

Part 2.2 Setup and Calibration of the pH Probes and Software

KEY INFORMATION
The head of the pH probe is incredibly sensitive. When using the probe, you
must:
• Ensure that the pH head is ALWAYS stored in solution when not in use. It must
not be allowed to dry out.
• Never rub the sensor tip. Instead, blot or dab the tip dry when changing solu-
tions.
• Never let any object hit the tip of the probe. This includes stir bars and the
bottoms of beakers and storage containers.

1. Obtain a Vernier GoDirect pH Sensor and connect it to your PC via Bluetooth or a USB cable.

2. Obtain a small amount of the pH 4.00, pH 7.00 and pH 10.00 buffer solutions from the chemical fume hood.

a Sometimes pH 8.00 is used instead of pH 7.00. This will work fine. Just make note of this.

3. Open the Vernier Graphical Analysis 4 software and choose “Sensor Data Collection.”

4. To calibrate the pH Sensor, click the “Sensor Actions” icon in the bottom right-hand corner of the software
window.

a Click “Calibrate.”
b Select a “Three-point calibration.”
c Carefully remove the probe AND the lid from the storage solution and slide the lid up the pH sensor.
d Carefully rinse with distilled water. Carefully blot the sensor head dry. Do not rub the head of the sensor
as it can damage it.
e Submerge the sensor into the pH 4.00 buffer solution. Wait for a constant voltage reading. Then enter
4.00 into the “First known value” box. Click “KEEP.”
f Carefully rinse and dry the sensor as before.
g Submerge the sensor into the pH 7.00 buffer solution. Wait for a constant voltage reading. Then enter
7.00 into the “Second known value” box. Click “KEEP.”
h Carefully rinse and dry the sensor as before.
i Submerge the sensor into the pH 10.00 buffer solution. Wait for a constant voltage reading. Then enter
10.00 into the “Third known value” box. Click “KEEP.”
j The probe should now be calibrated.

5. Adjust the data collection settings by clicking the “Data Collection Settings” icon on the bottom left side of

Laboratory Manual Prepared by Catalyst Education, LLC for University of North Carolina University at Chapel Hill BUS.8
Department of Chemistry.
 Part II: Preparation and Measuring the Buffer Capacity of Target Buffers

the software window.

a The “Mode” should be set to “Event Based.”

b The “Event Mode” should be set to “Events with Entry.”
c The “Event Name” should be entered as “Volume of Strong Acid/Base Added.”
d The “Units” should be “mL.”

Your probe and software should be ready to use.

Part 2.3: Preparation of the Target Buffers

1. Preparation of the Theoretical Buffer. Using the procedure you and your partner developed during step
3 of section 2.1, prepare a buffer by the theoretical method. Be sure to record all pertinent information
(masses, volumes, and pH of buffer).

2. Preparation of the Practical Buffer. Using the procedure you and your partner developed during step 4
of section 2.1, prepare a buffer by the practical method. Be sure to record all pertinent information (masses,
volumes, and pH of buffer).

Part 2.4: Measuring the Buffer Capacity of the Target Buffers

Select one of the two buffers prepared above to complete this section of the experiment.

Safety Precautions
CAUTION: You will be working with acids and bases which can chemically burn your skin. Additionally,
you must be careful not to add large amounts of your strong acid/base to the beaker.

KEY STEP
If possible, you should be using the same micropipettes that you calibrated last week! If you did not calibrate
micropipettes last week, please see your TA for additional guidance

1. Obtain a clean, 100-mL beaker and a stir bar. It is recommended that you take the time to clean both with
soap and water, and to rinse it with DI water thoroughly.

2. Fill the beaker with 20 mL of either the buffer prepared by the theoretical method or the practical method.
Place the stir bar into the beaker onto a stir plate.

3. Solutions of 0.100 M sodium hydroxide and 0.100 M hydrochloric acid were prepared for you. Be sure to
record the exact concentration of each in your laboratory notebook.

Laboratory Manual Prepared by Catalyst Education, LLC for University of North Carolina University at Chapel Hill BUS.9
Department of Chemistry.
 Part II: Preparation and Measuring the Buffer Capacity of Target Buffers

4. Obtain your calibrated micropipette and a small beaker with 0.1 M hydrochloric acid and 0.1 M sodium hy-
droxide.

5. Rinse, dry, and clamp the pH probe so that the tip is submerged in the solution and the stir bar can move
freely. If you need help, consult with your TA.

6. Start the stirring. Be sure to adjust the speed so that no material is splashed onto the side of the beaker.

7. Record the pH reading before the addition of any acid/base in the Vernier software by following the steps
below:

a Click “COLLECT.”
b Wait for a constant pH reading.
c Click “KEEP.”
d Enter the 0.0 mL added in the beaker and then click “KEEP POINT.”

8. SLOWLY, add the 0.1 M sodium hydroxide in 0.5-mL increments. Be sure to record the volume added and
the pH by following step 7 after each 0.5 mL added. Once you have reached pH 12 you may stop. If you have
added 25 mL or more, please consult with your TA.

9. It is recommended that you save the data as you go. To do this, click the “File Menu” icon at the top left of
the software window. Click “Save.” If this is your first time saving the data, you will be prompted to choose
a file name for the experiment. This will save the data in an .ambl format that can be read by the Vernier
Graphical Analysis 4 software. Once you are finished, export the data as a .csv.

a To export as a .csv file, click the “Page” icon. Then, click “Export” followed by “.CSV.” Choose a file
name for the experiment.

10. Once finished, dispose of the solution in the liquid waste container. Clean and dry the beaker. Refill it with
the same buffer you used before and repeat steps 4–9 by adding 0.100 M hydrochloric acid instead of sodium
hydroxide until the pH of solution is 2.00.

11. Once finished, dispose of the solution in the liquid waste container.

12. If there is time, you and your partner should repeat the steps above with the other buffer.

Laboratory Manual Prepared by Catalyst Education, LLC for University of North Carolina University at Chapel Hill BUS.10
Department of Chemistry.
 Part II: Preparation and Measuring the Buffer Capacity of Target Buffers

Part 2.5: Waste Disposal and Cleanup

1. Empty any remaining solutions into your waste beaker.

2. Empty any liquid waste into the liquid waste container. Place any dirty Kimwipes and weigh paper into the
solid waste container.

3. Return the pH sensors to the docking station at the front of the room. MAKE SURE that the probe is con-
nected and charging. When docked correctly, a blue LED will turn on to indicate that the probe is charging.

4. Return the 100-mL volumetric flasks and the micropipettes to the appropriate locations at the front of the
room.

5. Clean, rinse, dry, and return all glassware to the appropriate location.

Data Analysis

For this experiment, an Excel template (without the data and formulas) and a tutorial video are provided. The
video, which is posted under the Panopto tab on Canvas, will walk you through all steps of the analysis as well as
teach you tips and tricks for Excel. If at any point you need help with conceptual information, consult your TA. If
you need help with the use of Microsoft Excel, consult the tutorial video or the Internet. There are NUMEROUS
helpful how-tos online to complete any task in Excel.

The data analysis information provided for this experiment is more detailed than what will be provided for any
other experiment. You will be expected to rely on your conceptual knowledge from the lecture moving forward.

Reference

Laboratory Manual Prepared by Catalyst Education, LLC for University of North Carolina University at Chapel Hill BUS.11
Department of Chemistry.