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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

 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:

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.

 Acid-Base Pair pH of “Buffer” pH with 0.001 mol HCl added pH with 0.001 mol NaOH added HNO 3 and NaNO 3 1.30 1.25 1.34 HC 2 H 3 O 2 and NaC 2 H 3 O 2 4.74 4.65 4.83 NH 4 Cl and NH 3 9.25 9.16 9.34
 Increase Molarity 10X HNO 3 and NaNO 3 0.30 0.29 0.30 HC 2 H 3 O 2 and NaC 2 H 3 O 2 4.74 4.73 4.75 NH 4 Cl and NH 3 9.25 9.24 9.26 pH with 0.011 mol HCl added pH with 0.011 mol NaOH added HNO 3 and NaNO 3 0.30 0.25 0.35 HC 2 H 3 O 2 and NaC 2 H 3 O 2 4.74 4.64 4.84 NH 4 Cl and NH 3 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 HC 2 H 3 O 2 (0.10 M), with a K a value of 1.8 x 10 -5 . What is the pH? What does the addition of NaC 2 H 3 O 2 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) + H 2 O (l) H 3 O + (aq) + A - (aq), and when the weak acid is HC 2 H 3 O 2 , the

equilibrium equation would look like this: HC 2 H 3 O 2 (aq) + H 2 O (l) H 3 O + (aq)

+ C 2 H 3 O 2 - (aq). Based on this equation, we can set up an ICE table to calculate

the concentration of H 3 O + in the solution, and the calculated pH would equal to 2.89. The addition of NaC 2 H 3 O 2 would increase the concentration of C 2 H 3 O 2 -

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 NaC 2 H 3 O 2 is added.

b. How do the pK a values for acetic acid and ammonium ion (K a = 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 pK a 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 (HNO 3 ) in the first buffer is strong acid, and NaC 2 H 3 O 2 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) + H 3 O + (aq) --> H 2 O(l) + HA(aq)

b. The strong base, NaOH OH - (aq) + HA(aq) --> H 2 O(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 (Na 2 HC 6 H 5 O 7 ) and sodium citrate (Na 3 C 6 H 5 O 7 ), 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 (Na 2 HC 6 H 5 O 7 ) and sodium citrate (Na 3 C 6 H 5 O 7 ), with a net-ionic equation of: Na 2 HC 6 H 5 O 7 (aq) + H 2 O (l) H 3 O + (aq) + Na 3 C 6 H 5 O 7 (aq) . But since there is only citric acid (H 3 C 6 H 5 O 7 ) and not sodium monohydrogen citrate (Na 2 HC 6 H 5 O 7 ) and sodium citrate (Na 3 C 6 H 5 O 7 ) to start with, three steps of chemical reactions should be completed.

Step 1: H 3 C 6 H 5 O 7 + NaOH NaH 2 C 6 H 5 O 7 + H 2 O

Start with 100mL of 0.1M citric acid to obtain sodium dihydrogen citrate (NaH 2 C 6 H 5 O 7 ). Add 50mL of 0.2 M NaOH and completely react citric acid and obtain 150mL of sodium dihydrogen citrate (NaH 2 C 6 H 5 O 7 ).

Step 2: NaH 2 C 6 H 5 O 7 + NaOH Na 2 HC 6 H 5 O 7 + H 2 O

Then, to 150mL of sodium dihydrogen citrate (NaH 2 C 6 H 5 O 7 ), add 50mL of NaOH and completely react sodium dihydrogen citrate and obtain 200 mL of sodium bicitrate (Na 2 HC 6 H 5 O 7 ).

Step 3: Na 2 HC 6 H 5 O 7 + NaOH Na 3 C 6 H 5 O 7 + H 2 O

Finally, to 200mL of sodium bicitrate (Na 2 HC 6 H 5 O 7 ), add 25mL of NaOH so that half of sodium bicitrate (Na 2 HC 6 H 5 O 7 ) will react to form sodium citrate (Na 3 C 6 H 5 O 7 ).

The buffer solution obtained from these steps will contain equal concentrations of sodium bicitrate (Na 2 HC 6 H 5 O 7 ) and sodium citrate (Na 3 C 6 H 5 O 7 ).

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.

Data and calculations

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

pH
13
12.5
12
11.5
11
10.5
9.5 10 9
8.5 8
7.5
6.5 7
6
5.5 5
pH
0.035714286
0.785714286
1.535714286
2.285714286
3.035714286
3.785714286
4.535714286
5.285714286
6.035714286
6.785714286
7.535714286
8.285714286
9.035714286
9.785714286
10.53571429
11.28571429
12.03571429
12.78571429
13.53571429
14.28571429
15.03571429
15.78571429
16.53571429
pH
6.2
6
5.8
5.6
5.4
5.2
5
4.8
4.6
pH
0
0.785714286
1.571428571
2.357142857
3.142857143
3.928571429
4.714285714
5.5
6.285714286
7.071428571
7.857142857
8.642857143
9.428571429
10.21428571
11
11.78571429
12.57142857
13.35714286
14.14285714
14.92857143
15.71428571
16.5
17.28571429

Buffer: Sodium monohydrogen citrate (Na 2 HC 6 H 5 O 7 ) & Sodium citrate (Na 3 C 6 H 5 O 7 ).

2.

Calculate the anticipated pH for the buffer you were assigned. Include neat

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 (Na 2 HC 6 H 5 O 7 ) and sodium citrate (Na 3 C 6 H 5 O 7 ), and the general weak acid equation would look like this:

Na 2 HC 6 H 5 O 7 (aq) + H 2 O (l) H 3 O + (aq) + Na 3 C 6 H 5 O 7 (aq) .

Thus,

= [+][] []

.

We expect [HA] to be equal to [A-]; those two will cancel out, and Ka = [H 3 O + ].

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: Na 2 HC 6 H 5 O 7 (aq) + OH - (aq) Na 3 C 6 H 5 O 7 (aq) + H 2 O (l)

Adding Acid: Na 3 C 6 H 5 O 7 (aq) + H 3 O + (aq) Na 3 HC6H5O7 (aq) + H 2 O (l)

Our buffer was able to neutralize 5.5 mL of 0.2M NaOH and about 18 mL of 0.2M HCl.

NaOH: 0.2

1

1

∗ 5.5 = 1.1 x 10 -3 moles NaOH.

1000

HCl:

0.2

1

∗ 18 = 3.6 x 10 -3 moles HCl.

1

1000

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: Na 3 C 6 H 5 O 7 (aq) + H 3 O + (aq) Na 3 HC6H5O7 (aq) + H 2 O (l)

Molarity of Na 3 C 6 H 5 O 7 = 0.10 M

Moles of Na 3 C 6 H 5 O 7 =

0.10

1

1

1000

∗ 50 =

5.0 x 10 -3 .

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: Na 2 HC 6 H 5 O 7 (aq) + OH - (aq) Na 3 C 6 H 5 O 7 (aq) + H 2 O (l)

Molarity of Na 3 HC 6 H 5 O 7 = 0.10 M

Moles of Na 3 HC 6 H 5 O 7 =

0.10

1

1000 50 =

1

5.0 x 10 -3 .

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) + H 2 O (l)

The addition of water will shift the equilibrium to the right and increase

the concentration of the conjugate base, which will increase the pH.

H 3 O + (aq) + A - (aq) .

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 - + H 2 O

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,

= + ([

[−]

[] )

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 - + H 3 O + HCOOH + H 2 O 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.