In: Chemistry
A common biological buffer is a “phosphate buffer,” which involves the equilibrium between H2PO4- and HPO42-, with a pKa at 25 Celcius of 6.86. Describe in words how you would prepare 500. mL of 0.250 M phosphate buffer at pH = 7.2, given appropriate glassware and 500. mL each of 0.250 M NaH2PO4 and 0.250 M Na2HPO4 at 25 Celcius. (Note: your explanation needs to include actual volumes. It will be helpful to know that the concentrations of each component must add to the buffer concentration. i.e. [A-] + [HA] = 0.250 M)
pH = pKa + log ([A-]/[AH])
We have pH = 7.2 , pKa = 6.86
and [A-] + [AH] = 0.250 M .....1
therefore,
7.2 = 6.86 + log ([A-]/[AH])
log ([A-]/[AH]) = 0.34
([A-]/[AH]) = 2.188
[A-] = 2.188 [HA]
from equation 1.
2.188 [HA] + [HA] = 0.25 M
[HA] = 0.25/3.188 = 0.078 M
[A-] = 0.250-0.078 = 0.172 M
We need H2PO4- 0.078 M of 500 mL solution. We have 0.250 M, so we need to calculate the volume we need to take.
M1V1= M2V2
0.078 M x 500 mL = 0.25 M x V2
V2 = 156 mL
Similarly, we need HPO42- 0.172 M of 500 ml solution. We have 0.250 M, so we need to calculate the volume we need to take.
M1V1= M2V2
0.172 M x 500 mL = 0.25 M x V2
V2 = 344 mL
Therefore, we need to mix 344 mL of 0.250M of NaH2PO4 and 156 mL of
0.250M Na2HPO4 to make (344 +156 = 500) 500. mL of 0.250 M
phosphate buffer at pH = 7.2