Theory:

A buffer solution is one that is resistant to change in pH when small amounts of strong acid or base are added. For
example, when 0.01 mole of strong acid or base are added to distilled water, the pH drops to 2 with the acid and rises to 12
with the base. If the same amount of acid or base is added to an acetic acid – sodium acetate buffer, the pH may only change a
fraction of a unit.
Many buffers are prepared by combining a weak acid and its conjugate (acetic acid and sodium acetate) or a weak
base and its conjugate (ammonia and ammonium chloride). In general, the pH range in which a buffer solution is effective is +/-
one pH unit on either side of the pKa. The Henderson Hasselback equation:

𝑝𝐻 = 𝑝𝐾𝑎 + log conj𝑢𝑔𝑎𝑡𝑒 𝑏𝑎𝑠𝑒 (𝐴)

𝑤𝑒𝑎𝑘 𝑎𝑐𝑖𝑑 (𝐻𝐴)
There is a limit to the amount of acid or base that can be added to a buffer solution before one of the components is
used up. This limit is called the buffer capacity and is defined as the moles of acid or base necessary to change the pH of one
liter of solution by one unit.

𝐵𝑢𝑓𝑓𝑒𝑟 𝐶𝑎𝑝𝑎𝑐𝑖𝑡𝑦 = 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝑂𝐻− 𝑜𝑟 𝐻3𝑂 +

(𝑝𝐻 𝑐ℎ𝑎𝑛𝑔𝑒)(𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑏𝑢𝑓𝑓𝑒𝑟 𝑖𝑛 𝐿)

Materials:

100and 400 mL glass jars 0.10 M CH3COOH

0.02 M NaOH (soap) NaHCO3
Pipette/dropper NaCO3
pH Meter (in the internet) Bottle

Sample Computation:

How would you prepare 10mL of a 0.01M phosphate buffer, pH 7.40, from stock solutions of 0.10M KH2PO4and 0.25M
K2HPO4? pKa of KH2PO4 = 7.20. Prepare 10 mL of a 0.01 M phosphate buffer, pH 7.70, from stock solutions of 0.1 M
K2HPO4and 0.2 M KH2PO4.
(pKa for the weak acid = 7.20).

1. Use the Henderson Hasselbalch equation to find the ratio of A

pH = pKa + log [A-] / [HA]
7.40 = 7.20 + log [A-] / [HA]
0.20 = log [A-] / [HA]
1.584893192 = [A-] / [HA]*

*Since [A-] / [HA] = 1.584893192, we can say that [A-] / [HA] = 1.584893192/1. In this case
[A-] = 1.584893192; [HA] = 1.

2. Calculate the decimal fraction (part/whole) of each buffer component.

A- = 1.584893192 / (1.000 + 1.584893192)
=1.584893192/2.584893192 = 0.61313682
HA = 1.000 / 2.584893192 = 0.38686318

3. Find the molarity (M) of each component in the buffer by simply multiplying the molarity of the buffer by the decimal fraction
of each component.
MA- = 0.01M x 0.61313682 = 0.006131368M
MHA = 0.01M x 0.38686318 = 0.003868632M

4. Calculate the moles of each component in the buffer.

Moles = Molarity x Liters of buffer
moles A- = 0.006131368M x 0.01L = 6.131 x 10-5
moles HA = 0.003868632M x 0.01L = 3.869 x 10-5

5. Calculate the volume of each stock solution required to make the buffer
Liters of stock = moles of the buffer component / Molarity of the stock
LA- = 6.131 x 10-5 moles / 0.25 M = 2.452 x 10-4 L = 245
Liters of stock = moles of the buffer component / Molarity of the stock
LA- = 6.131 x 10-5 moles / 0.25 M = 2.452 x 10-4 L = 245µL
LHA = 3.869 x 10-5 moles / 0.10 M = 3.86 9 x 10-4 L = 387 µL
Procedure:

A. Phosphate Buffer

1. Prepare 100 mL of a 0.01 M phosphate buffer, pH 7.70, from stock solutions of

0.1 M K2HPO4and 0.2 M KH2PO4. (pKa for the weak acid = 7.20).

a. Use the Henderson-Hasselbalch equation to calculate the volume of each stock

solution needed.
pH = pKa + log [conjugate base] / [weak acid]
b. Check your calculations with other students. See the instructor if there is
uncertainty.
c. Make the solution and check the pH of a portion of your buffer solution using the
pH meter.

B. Acetate Buffer

2. Prepare 100 mL of 0.01 M acetate buffer,pH 3.80, from stock solutions of 0.1 M
acetic acid and 0.02 M sodium hydroxide. pKa acetic acid = 4.76.

a. Use the Henderson-Hasselbalch equation to calculate the volume of each stock solution needed.
pH = pKa + log [conjugate base] / [weak acid]

b. Check your calculations with other students. See the instructor if there is uncertainty.
c. Make the solution and check the pH of a portion of your buffer solution using the pH
meter.
d. Calculate the exact volume of the 0.01 M acetate buffer required to make 100 mL of
0.0005 M acetate buffer.
e. Prepare this new buffer using the following equation to aid you in your calculations.
C1V1 = C2V2
d. Check the pH of this new buffer.

C. Bicarbonate Buffer

3. Prepare 100 mL of 0.200 bicarbonate buffer, pH 10 by calculating the mass of sodium

bicarbonate and sodium carbonate.

a. Use the Henderson-Hasselbalch equation to calculate the volume of each stock

solution needed.
pH = pKa + log [conjugate base] / [weak acid]1
b. Check your calculations with other students. See the instructor if there is uncertainty.
c. Make the solution and check the pH of a portion of your buffer solution using the pH
meter.
L = 245μL
DATA:

Show your calculation:

1. Preparation of Phosphate Buffer

Calculated Volume
K2HPO4 _________________________________
KH2PO4 _________________________________
Actual pH of the buffer: ____________________

2. Preparation of Acetate Buffer

Calculated Volume
CH3COOH _________________________________
KH2PO4 _________________________________

Actual pH of the buffer: ____________________

3. Preparation of Bicarbonate Buffer

Calculated Volume
CH3COOH
KH2PO4

Actual pH of the buffer: ____________________