Buffers

Alex Johnson, Jonathan Taylor

Introduction: Buffers are solutions that are able to resist a ph change when encountered with a strong
acid or base. Buffers are usually made by adding a weak acid and its conjugate base to a solution. In this
lab we are not using the Henderson Hasselbalch equation to calculate the amounts of each substance
needed to make the buffer. Instead we are weighing out the correct amounts of each and adding water
and titrating the solution until we reach the desired ph. To measure ph we are using a ph electrode.

Procedure: To make 50mL of 1.0M Tris buffer at a ph of 6.8, 6.057g of tris was weighed and placed into a
150mL beaker. 30mL of dh2o was added to the beaker and the solution was mixed with a glass stirring
rod until it was clear. Then the pH meter was placed into the solution and it was noted that the solution
was far too basic. To bring the pH down to 6.8, 170 drops of HCl were added. The solution was
transferred to a 50mL volumetric flask and water was added to the solution to bring the final volume to
50mL. To make 50mL of 1.5M Tris buffer at a ph of 8.8, 9.086g of Tris was weighed and placed into a
150mL beaker. 30mL of dh2o was added to the beaker and the solution was mixed with a glass stirring
rod until it was clear. Then the pH meter was placed into the solution and it was noted that the solution
was far too basic. To bring the pH down to 8.8, 50 drops of HCl were added. The solution was
transferred to a 50mL volumetric flask and water was added to the solution to bring the final volume to
50mL.

Data and results:

50mLx (1L/1000mL) x (1mol/1L) x(121.14g/1mol)= 6.057g Tris

50mLx (1L/1000mL) x (1.5mol/1L) x(121.14g/1mol)= 9.086g Tris

Discussion: In this lab I learned the practical way to prepare a buffer. I experienced firsthand the
capacity for buffers to resist changes in pH when an acid is added. In this lab I also learned how to
calibrate and use an electronic pH meter.

Lab Questions:

1. 7.00 =7.21 + log (A-/HA)

-.21= log (A-/HA)

.617= (A-/HA) A-+HA=.3M A-= .3M-HA

.617= (.3M-HA)/HA)

HA=.186M A-= .114M

.186Mx 121g/mol = 22.5g NaH2PO4

.114Mx 142g/mol = 16.2g Na2HPO4

 Add 22.5g NaH2PO4 and 16.2g Na2HPO4 to a beaker and then fill the beaker to 1L.

2. HEPES (pKa = 7.55), MES (pKa = 6.15) TRIS (pKa=8.06) Use MES to make a solution with a ph of
6.0.
3. The buffer capacity of a solution is the ratio of incremental amounts of base added to the
corresponding change in ph.

HO
HO
HO HO
NH2 NH–
4. HO HO