University of Texas Rio Grande Valley

Week Six: pH and Buffers

By: Gillian Mendez

BMED 1102-1BR
Dr. Seratna Guadarrama

Introduction:
In this lab, students will familiarize themselves with the stages of the pH scale and how
solutions react to the addition of acids or bases. They will practice creating “buffers”, or
solutions that typically resist change in pH (Helmenstein) in order to better understand how they
are made and how they work. These experiments allow for students to get a more visual

Objective:
Students will use Phenol Red (a known pH indicator, along with Biomythymol blue, to provide a
visual determination of changes in pH. They will also create an active buffer, and a base pH, as
well as measure the differing pH of solutions once exposed to HCl.

Materials, Experiment Setup, and Procedure:

For the first activity, students used
● 3 125mL flasks
● 3 straws
● A timer
● Phenol red solution.
Students added 50 mL of ddH20 and then 1mL phenol red. Using a straw, students would blow
until the solution changed colors, noting the time it took to do so.

For the second activity or making an Acetate Buffer, students used

● .2m Acetic Acid
● .2m Sodium Acetate
● 50mL conical tubes
● Electrical pipette
● 10mL serological pipette
● Ph meter
● Tube racks
Using five 50mL conical tubes, labelled 0-4, students would add sodium acetate and/or acetic
acid. Below, an image of properly labelled and filled tubes has been attached to demonstrate
the correct setup.
Activity Three was fairly similar to Activity Two. It required:
● Acetic Acid
● Sodium Acetate
● HCl
● NaOh
● ddH20
● 50mL conical tubes
● Pipettes
● Serological pipette
● Electric pipette
● pH meter
The procedure was as follows: label one 5mL tube “buffer” and another “water”. In buffer, add
10mL acetic acid and 10mL sodium acetate. In water, add 20mL pure water. Measure the pH of
both. Then, add one drop HCL and check pH. Repeat 4 more times.

Below, a picture demonstrates the proper way to use an electronic pH measure.

Results and Data:
Unfortunately, our group was the one that received discolored Phenol Red. We noticed too late
to receive a fresh batch but still proceeded with the experiment. Our phenol red, as shown
below, was much more orange in color, although after 3 minutes of blowing, we noticed a shift
from a dark orange to a lighter color.
A table showing the pH results for the Acetate Buffer solution has been included below.

Tube 0 5.47

Tube 1 5.28

Tube 2 5.41

Tube 3 4.80

Tube 4 4.52

Here is a coherent table noting the pH of both a buffer and water after the addition of HCL. This
table is for Acid pH

ADDITIONS pH of Buffer pH of Water

NONE 4.85 8.55

5 drops of HCl 4.95 2.65

10 drops of HCl 4.87 2.30

15 drops of HCl 4.82 2.15

 20 drops of HCl 4.78 1.96

25 drops of HCl 4.72 1.89

Note: The buffer, as per design, is obviously resisting a change in pH. Despite the addition of 25
drops of Hydrochloric acid,the pH does not differ very much. The strongest change, is less than
.40. This is a sharp contrast to the water which, beginning with a fairly neutral pH of 8.55, drops
to a 2 on the pH scale after the addition of 5 drops of HCl. After 25 drops have been added, the
water has transformed to a fairly strong acid.

The following table is for Base pH:

ADDITIONS pH of Buffers pH of Water

NONE 4.85 8.55

5 drops of NaOH 4.98 11.88

10 drops of NaOH 5.07 12.17

15 drops of NaOH 5.12 12.37

20 drops of NaOH 5.16 12.43

25 drops of NaOH 5.20 12.56

The results here show the change in pH after sodium hydroxide (NaOH), a very strong base, is
added to both a buffer and a conical of water. As demonstrated in the previous experiment, the
buffer resists change in pH. Over the course of 25 drops of NaOH, it only becomes a slightly
stronger base. Contrarywise, water again is shown to readily change pH. After 25 drops, it has
become a very strong base with a pH of 12.56. Compared to its starting pH of 8.55, this is a
drastic change.

Discussion and Conclusion:

With the information that has been gathered gathered, students can conclude that no matter
how strong the addition, a buffer will always resist change in pH. Both of the Acid pH and Base
pH activities show how readily water will adapt to a new pH, often changing by several points
after only a few drops of NaOH or HCl.

Again, the Phenol Red did not react as hoped, although by noting the slight color change, it can
be concluded that the carbon dioxide created when a person exhales is enough to dissolve the
water in the flask, thus creating a carbonic acid and lowering the pH. Had the Phenol Red not
been off-color, the change would have been more drastic and easier to note.
References:
Helmenstine, P. A. (n.d.). What Buffers Are and How They Work. Retrieved October 07, 2017,
from https://www.thoughtco.com/buffers-in-acid-based-chemistry-603647