How the Power of Hydrogen Works: Buffer Testing and Colorimetric

Determination

Cruz, A., Dela Vega, E., Dizon, A., Esguerra, M.J., Espiritu, M.A., Garcia, E.M.

Abstract

A buffer is a solution that resists changes in pH. It is made from the combination
of a weak acid and its salts. The experiment was done by preparing different buffer
solutions and achieved through the following: determining the pH of the buffers and
samples colorimetrically by using different liquid indicators and electrometrically by using
the pH meter. It was prepared using Glacial Acetic Acid (𝐶𝐻3 𝐶𝑂𝑂𝐻) as weak acid and
Sodium Acetate (𝐶2 𝐻3 𝑁𝑎𝑂2 ) as conjugate base. The amounts of buffer component were
calculated using the Henderson-Hasselbalch equation. A volume of 500mL of 6M HCl and
6M NaOH were prepared to adjust the buffer solution to its desired pH. The pH meter
showed accurate pH readings of the prepared buffer solution. The pH of the prepared
buffer solution was then measured colorimetrically using acid-base indicators such as
thymol blue, bromophenol blue, bromocresol green, bromocresol purple, methyl red,
methyl orange, phenolphthalein and phenol red. Colorimetric determination of the pH
showed the varying color changes an acid-base indicator undergoes when added to a
solution of certain pH. This property of an acid-base indicator can be used to identify
different substances by narrowing their pH range. It therefore can help in the
identification of a substance since different substances have different pH levels.

I. Introduction excess and the hydroxyl ion which makes

The term pH is the measure of the
hydrogen ion concentration in a given
aqueous solution. This was first
introduced by a Danish Chemist named
Peder Lauritz Sørensen in 1909. The
word pH is an abbreviation for the power
of hydrogen which “p” comes from the
word power in German which is potenz
and ‘H” for the element hydrogen. The H
is capitalized because it is a standard to
capitalized symbols of the element. A
buffer solution is used to prevent the
change of pH with the added solute.
They are resistant to pH change as a
result of the presence of an equilibrium
between the acid and its conjugate base.
The components neutralize the hydrogen
the aqueous solution resist large changes
in the pH. The buffer is important
because this contributes to help
maintain a close constant pH by adding
small amounts of hydrogen or hydroxide
to the given solution.
The Henderson-Hasselbalch
equation shows the relation of the pH of
the solution with the mixture of the two
components to the acid dissociation
constant or the Ka and the other species’
concentration in the solution. This is also
significant in determining if a solution of
a conjugate acid or base pair will function
as a buffer. In this experiment we
prepared different solutions of buffer to
help determine the buffer’s pH and the
samples colorimetrically with the help of
liquid indicators.
II. Methodology
A. Reagents
For preparation of acetate buffer
solution, the reagents of glacial acetic
acid and sodium acetate are used. For pH
analysis, the acid-base indicators are:
bromophenol blue, bromocresol blue,
bromocresol green, bromocresol purple,
phenol red, methyl red, methyl orange,
phenolphthalein and thymol blue. The
assigned sample is salicylic acid solution.
B. Procedure
I. Preparation of Reagents
In preparing the reagents, 500mL
of 6.0M of NaOH and 6.0M HCl is to be
used for the adjustment of pHs in the
buffer solution, obtaining the desired pH,
after being labeled properly and given to
different buffer solutions.
II. Preparation of Buffers
For the preparation of the buffer
solution, the group was assigned to
prepare a 50 mL 0.50 M acetate buffer
solution with the desired pH of 5.00 and
pKa = 4.70 and will be tested for its pH.
The weak acid is glacial acetic acid
(𝐶𝐻3 𝐶𝑂𝑂𝐻) and the conjugate base is
sodium acetate (𝐶2 𝐻3 𝑁𝑎𝑂2 ). In order to
prepare sodium acetate, NaOH pellets
(MM 40) and glacial acetic acid (97%
w/w, sp.gr. 1.05, MM 60) are computed
and mixed along with glacial acetic acid,
to create the acetate buffer solution. The
equation used to identify the portions of
the weak acid and conjugate base
needed for the preparation is shown
below:
pH = pKa + log ([A-]/[HA])
[A-] represents the molar concentration of a
conjugate base and the [HA] as the molar
concentration of an undissociated weak acid (M)
After measuring the components of
Weak Acid and Conjugate Base and
dissolved together to make a solution, it
is dissolved in a small amount (35mL) of
water. If the desired pH (5.00) was not
reached, use the 6.0M HCl or NaOH to for
adjustment of pH. Transferred to a 50mL
Volumetric flask, fill to volume and
labeled properly.
III. Electrometric Determination of pH
In order to measure the pH of a
200uL portion of the following samples:
Prepared Acetate Buffer solution,
Distilled Water and Assigned sample
(SA), the pH meter must be calibrated at
pH 4, 7 and 10 for accuracy of pH levels.
If the desired value for Acetate Buffer
solution is not achieved, the reagents of
6.0M HCl or 6.0M NaOH is added to the
prepared solution.
IV. Colorimetric Determination of pH
All the needed reagents were placed
in a microplate along with other buffer
solutions in diff. pKa & pH, distilled water
and assigned sample (Salicylic Acid
solution) in the amount of 200 uL paired
with the different Acid-Base indicators, in
the amount of 20uL. The assignment of
the microplate are:
Table 1.1 Assignment of pH and Acid-Base
Indicator
Indicators pH
A1 – A9 Thymol blue A1 – H1 pH 2.0
Bromophenol
B1 – B9 A2 – H2 pH 3.0
blue
 Bromocresol Table 1.2 pH concentration of distilled
C1 – C9 A3 – H3 pH 5.0 water, salicylic acid and the buffer
green
Bromocresol
D1 – D9 A4 – H4 pH 7.0
purple The table shows the pH of the distilled
E1 – E9 Methyl red A5 – H5 pH 7.5 water which is 7.0 and the concentration
of hydrogen is 1x10-7. Salicylic acid
F1 – F9 Methyl orange A6 – H6 pH 8.0 (sample) with the pH of 2.7 and ion
concentration of 1.10x10-3. Prepared
G1 – G9 Phenolphthalein A7 – A7 pH 12.0
buffer with the pH of 5.0 and
pH concentration of 1x10-5.
H1 – H9 Phenol red A8 – A8 Distilled
water
Salicylic To compute the concentration of
A9 – H9 hydrogen [𝐻 + ] the formula used is:
acid
[𝐻 + ]=10-pH

Colorimetric analysis
Table 1.3 The table shows how the acid-
base indicator reacted with the different
pH by producing colors.

Figure 1.1 & 1.2. Application of different pH &

Acid-Base Indicators

III. Results and Discussion

The pH and Concentratiom of

Distilled water, Sample and buffer
Figure 1.3 Reaction of the different pH by
producing colors.
Samples pH [H+]

Distilled Water 7.0 1x10-7

Sample (Salicylic 2.7 1.10x10 -3

Acid)

Buffer Prepared 5.0 1x10 -5

A. Electrometric Determination of pH
The calibrated electrometric pH
meter is used to determine an accurate
reading of the pH concentration of the
standard buffer solution. It showed
instability and change of concentration
on the slightest volume of 0.10M HCl or
0.10 M NaOH) added, until the desired
pH concentration is achieved. The group
adjusted the pH of the buffer from 5.4 to
its desired pH, 5.0.
B. Colorimetric analysis
In this experiment, colorimetric
analysis was used as a qualitative
method of analysis and determination of
the pH concentration of a chemical
substance through its color intensity. It
was seen in the results that with the
variety of pH concentration and acid-
base indicator suspended in the solution,
a change in color was observed. This will
be narrowed down to their corresponding
pH range. The change in color was due
to the emergence of a byproduct of the
added acid-base indicator or an intrinsic
property of the substance in the solution.
The effect of diluting the acid-
base indicator to the solution changes
the color thus the change of its hydrogen
ion concentration.
VI. Conclusion
In conclusion, the proper
calibration of an electrometric pH meter
is significant in getting an accurate pH
measurement of a buffer solution. With
regards to the colorimetric determination
of pH, it can be concluded that the
change of color in the addition of a
specific acid-base indicator to a buffer
solution relies on its pH level. Each
indicator has a specific color change that
would entail the pH range of a buffer.
VII. References
● Chemicool. Retrieved from
https://www.chemicool.com/definition/ph
.htmlhttps://courses.lumenlearning.com/
boundless-chemistry/chapter/buffer-
solutions
● Helmenstine, A. (2019). Henderson-
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from:https://www.thoughtco.com/hender
son-hasselbalch-equation-and-example-
603648W
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Report. Retrieved September 8, 2013,
from
Scribd.:http://www.scribd.com/doc/7759
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● Murray, R.K., Granner D.K., and Rodwell,
V.W. (2006). Harper’s llustrated
Biochemistry. 27th ed. Singapore:
McGraw-Hill Companies Inc. (Asia)
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● Wilson K., and Walker J. (2005).
Principles and Techniques of
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