Experiment 5: Examination of Buffer Solutions

Introduction
Buffer solutions contain a weak acid and its conjugate base, or a weak base and its conjugate acid, such that the resultant solution resists change in pH. In this experiment, you will explore properties of buffer solutions. In particular, you will investigate the impact of: 1) the relative concentrations of buffer components, 2) the addition of a strong acid or base, and 3) dilution on the pH of a buffer solution and/or its components. You will work with a partner for this experiment. You and your partner will examine either the acetic acid/acetate or the ammonia/ammonium buffer system.

Physical and Chemical Principles

The pH of a solution can affect both the spontaneity and the rate of chemical reactions. The ability to control the pH of a solution is, therefore, of fundamental importance. Because many cellular processes can result in the generation of acids or bases, organisms must have a way to regulate the pH of cellular and extracellular fluids. In general this is accomplished through the use of buffers, or solutions that resist a change in pH. The components of a buffer solution are a weak acid and its conjugate base, or a weak base and its conjugate acid. The identities of the buffer components and the relative amounts of the weak acid and conjugate base (or weak base and conjugate acid) dictate the pH of a buffer. This is most easily observed through the Henderson-Hasselbalch equation (equation 1), which relates the pH of a buffer solution to the pKa of the acid component and the concentrations of the weak acid and conjugate base (or weak base and conjugate acid). pH = pK a + log [base] [acid] (1)

It is important to note that the concentrations present in the Henderson-Hasselbalch equation represent the initial concentrations of the acid and base components of the buffer, without consideration of equilibrium effects. In the vernacular of equilibrium calculations, this means that the small x approximation is assumed to be valid for both components of the buffer. It is always important to check whether or not this assumption is valid when using the HendersonHasselbalch equation for quantitative pH calculations. This experiment seeks to examine various aspects of buffer solutions. Multiple buffer solutions for a particular system (either the acetic acid/acetate system or the ammonia/ammonium system) will be generated. By measuring the pH for these solutions, the validity of the HendersonHasselbalch equation can be gauged. Additionally, by adding strong acid and strong base to various solutions, the ability of buffer solutions to resist changes in pH relative to water will be assessed. Finally, the impact of dilution on the pH of solutions of buffer components will be examined. From these results, the effect of dilution on a buffer can be inferred.

Experimental
Table 1: Equipment and Chemicals for Examination of Buffer Solutions. Equipment Volumetric pipets, 2, 5, and 8 mL Volumetric pipets, 10 mL Volumetric flasks, 100 mL Assorted flasks and beakers pH meter Chemicals (Note: you will be assigned EITHER the acetic acid/acetate OR the ammonia/ammonium buffer) Standard buffer solutions (4 and 7 or 7 and 10) Acetic acid, 0.50 M Sodium acetate, 0.50 M Ammonia, 0.50 M Ammonium chloride, 0.50 M Hydrochloric acid, 0.10 M Sodium hydroxide, 0.10 M Deionized water Quantity 1 each 2 4 As needed 1 Quantity 1 vial each 200 mL 200 mL 200 mL 200 mL 50 mL 50 mL As needed

Preparation
Notebook Students are expected to arrive at lab prepared to start the experiment. The lab notebook should contain preliminary notes sufficient for any person to complete the experiment without the lab manual. Instructors and TAs will be checking notebooks to see that sufficient detail is present to complete the lab. Students that cannot demonstrate they are ready to start the experiment will be asked to leave the lab. Pre-lab assignment Students must prepare answers to the pre-laboratory worksheet before entering lab. Individuals without completed pre-laboratory assignments will not be allowed to participate in lab. Worksheets will be collected at the start of lab

Procedure
1. pH meter calibration a) Before using the pH meter, it must be calibrated using buffer solutions of known pH. Make sure that you or your partner has obtained vials of the appropriate buffers: if you are examining the acetic acid/acetate buffer system, you should calibrate the pH meter using the pH 4 and 7 solutions; if you are examining the ammonia/ammonium buffer system, calibrate the pH meter with the pH 7 and 10 solutions. b) Each pH meter will have a small card with instructions for the use of the pH meter. Ask your instructor or TA if you cannot locate this instruction card. Often, the electrode for the pH meter will have parafilm or tape covering a small hole in the top of the electrode. Be sure to remove this parafilm and/or tape and make sure that this opening is exposed to air before calibrating the pH meter. c) Carefully remove the electrode from the electrode storage solution. Rinse the electrode with deionized water and carefully remove the excess water with a Kimwipe or other lintfree tissue. Submerge the electrode approximately one-half to one inch into a buffer of pH 4 (or 7). Follow the instructions for the pH meter in order to calibrate the pH meter for this solution. Follow a similar process to provide a second point of calibration using the pH 7 (or 10) buffer (be sure to rinse and dry the pH electrode before placing it in the new buffer solution). If the pH meter gives you an error message or does not provide a stabilized reading, ask your instructor or TA for help. d) Be careful with the pH electrode. You should not use the electrode to stir the solution. It is also bad for the electrode to dry out; therefore, the electrode should remain in some solution throughout the experiment. You should always rinse the pH electrode with deionized water and pat it dry with a Kimwipe before using it to determine the pH of a solution. 2. Preparation and pH measurement of buffer solutions a) Obtain approximately 200 mL each of the 0.50 M solutions of the weak acid and weak base that you have been assigned (either acetic acid and sodium acetate or ammonia and ammonium chloride). In addition, obtain approximately 50 mL each of the 0.10 M solutions of HCl and NaOH. Be sure to record the actual concentrations of all of the solutions. b) Using the 2, 5, 8, and 10 mL volumetric pipets, dispense the volumes listed in Table 2 (see the next page) into beakers or Erlenmeyer flasks. Be sure to label each beaker/flask with the appropriate designation. c) After ensuring that the solutions are well mixed, measure and record the pH of each of the solutions prepared in part (b). After measuring the pH of all of the solutions once, return to the first solution and record a second measurement of the pH of each solution. Your two measurements should agree within 0.1 pH units. If this is true for your pH

meter, you can just make one pH reading for all other solutions examined in this experiment. d) Keep solutions A1, C1, and D1, as you will need them for the next part of the experiment. Table 2: Volumes Needed to Generate Various Buffer Solutions Solution Volume of acetic acid OR Volume of sodium acetate ammonium chloride (mL) OR ammonia (mL) A1 10 10 B1 C1 D1 E1 15 18 2 5 5 2 18 15

3. Examination of the effect of the addition of a strong acid/base a) Using the volumes given in Table 2, generate a solution identical to A1 (label it A2), a solution identical to C1 (label it C2), and a solution identical to D1 (label it D2). Finally, add 20 mL of deionized water (using a volumetric pipet) to each of two clean and dry beakers or flasks. Label the beakers/flasks Water 1 and Water 2. Measure and record the pH of each solution. b) Add 10 mL of 0.10 M HCl to solutions A1, C1, D1, and Water 1. Add 10 mL of 0.10 M NaOH to solutions A2, C2, F2, and Water 2. After ensuring that the solutions are well mixed, measure and record the pH of each solution. 4. Examination of the effect of dilution a) Determine the pH of the 0.50 M acetic acid and sodium acetate (or ammonia and ammonium chloride) solutions. b) Using a volumetric pipet, add 10 mL of the 0.50 M acetic acid (or ammonium chloride) solution to a 100 mL volumetric flask and fill to the mark to make a 1:10 diluted solution. Mix this solution and pour it into a clean and dry beaker or flask. Do the same for the 0.50 M sodium acetate (or ammonia) solution. Be sure to label the beakers/flasks. c) Add 10 mL of each of the 1:10 diluted solutions from part 4b to a new 100 mL flask and fill each one to the mark to generate 1:100 diluted solutions. Mix these solutions and pour them into clean and dry beakers or flasks. Be sure to label the beakers/flasks. d) Measure and record the pH of the 1:10 and 1:100 diluted solutions.

Waste Disposal
Place the contents of all beakers and flasks in the aqueous waste containers provided in the lab.

Calculations
This section describes the calculations that you will need to perform in order to successfully complete your laboratory assignment. Read through them carefully and make sure you understand them all before you leave the laboratory. Preparation and pH measurement of buffer solutions 1. Calculate the logarithm of the ratio of the concentration of sodium acetate to acetic acid or ammonia to ammonium chloride, i.e., find log([base]/[acid]), for each of the solutions listed in Table 2. Be sure to use the actual concentrations of the solutions given to you in the laboratory. 2. Using the values from the step above, generate a plot of pH versus log([base]/[acid]). Fit the data to a linear curve Examination of the effect of the addition of a strong acid/base 1. Calculate the theoretical pH of solution A1 (which is identical to that of solution A2). In addition, calculate theoretical values for the pH of the solutions obtained after adding 10.00 mL of 0.10 M HCl to solution A1 and 10.00 mL of 0.10 M NaOH to solution A2. Be sure to use the actual concentrations of HCl and NaOH from the lab. Examination of the effect of dilution 1. Calculate the pH expected for each of the solutions examined in procedure step 4 (six total calculations). Be sure to check that any simplifying assumptions made are valid. You only need to show one sample for each type of calculation performed. 2. Make a table that compares the actual pH values obtained to the calculated pH values. 3. Plot the experimentally obtained pH values versus the dilution factor (1, 10, and 100).

Assignment
The report for this experiment is due at the beginning of the next laboratory period. It must be neatly typewritten and contain the following elements: The report cover sheet available from the course web site. Purpose Write a paragraph consisting of several sentences that describe the nature and goals of the experiment. This section should concisely answer the following questions: What data are being collected during the lab? How are the data being collected? How are the data used to reach a result, or what is the relationship between the recorded data and the calculated value of interest?

What concepts and techniques are illustrated in the experiment? Spelling and grammar must be correct to receive full credit for this section. Procedure It is sufficient to cite the procedure provided above (remember to provide a suitable reference or references if you used additional sources). Note any changes to the procedure in a bulleted list. If there were no changes made to the experimental procedure, this should be noted. Even in this instance, it may be that the lab did not proceed as expected. Comments on unexpected events can be stated here in a bulleted list. Data and Results Data tables 1. Create a table containing the pH values for each of the buffer solutions examined in procedure step 2. 2. Make a table that reports the experimental pH values recorded for solutions A1, C1, D1, Water 1, A2, C2, D2, and Water 2. Have additional columns in which the pH values of the solutions after the addition of acid or base are recorded. Finally, have a column in which the pH differences (pH after acid or base initial pH) are recorded. The sign of the pH should make sense, i.e., after the addition of acid, the pH should decrease, leading to a negative pH difference. A suggested format for the table is shown below. Table 3: Suggested format for the Effect of Adding Acid or Base Solution A1 C1 (continue for other solutions) Initial pH pH after adding HCl pH after adding NaOH pH Difference

3. Create a table that compares the calculated and actual values of the pH of the initial and diluted solutions for procedure step 4. Plot of pH vs. log([base]/[acid]) Plot the pH of each of the solutions generated in procedure step 2 vs. the logarithm of the ratio of the concentration of the base component of the buffer to the concentration of the acid component. Report the equation of the best-fit line to the data in a figure caption below this plot.

Plot of pH vs. concentration Plot the pH of the solutions examined in procedure step 4 vs. the dilution factor (1, 10, 100) Sample calculations In a step-by-step fashion show each calculation step for the lab. Provide a brief description of each step in your own words. In general, equations should be written first using symbols, followed by a sample calculation using experimental values. When equations are displayed in written work, they should be centered on the page. The sample calculations may either be typewritten or hand-written. In either case, the work must be well organized and neat. If the instructor cannot read, or follow the sample calculations, no credit will be awarded to this section of the report. Discussion Discussion sections should be presented in paragraph form, not simply numbered answers to the questions listed below. The questions provided should be answered in an overall discussion that describes the quality of the results obtained and states any possible sources of error in the experiment. Part of your discussion grade will be based on the clarity of the writing in and the overall flow of this section. 1. State the values you would have expected to obtain for the slope and yintercept of the best-fit to line to a plot of pH vs. log([base]/[acid]) for your buffer system. Compare your expected values to those actually obtained and provide a possible explanation for any differences observed. 2. Comment on the relative pH difference values observed in procedure step 2. Do all of the buffer solutions show the same change in pH on the addition of an identical amount of acid or base? Why or why not? In addition, compare the pH difference values obtained for water to those obtained for the buffers. Finally, compare calculated pH values after adding acid and base to buffer A1 to the experimental values obtained. Provide an explanation for any discrepancies observed. 3. Does the pH of the acetic acid (or ammonium chloride) solution increase or decrease as a result of dilution? Answer the same question for sodium acetate (or ammonia). Explain your answers. In addition, predict whether you would expect the effect of dilution on the pH of a solution of HCl to be greater or less than that observed for a solution of acetic acid (or ammonium chloride). Answer the same question for a buffer solution such as buffer A1 in your experiment. Explain your answers.