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 (CH3COOH) as weak acid and
Sodium Acetate (C2H3NaO2) 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 known to be 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
equations 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 to use 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 [𝐻𝐴/𝐴] = 𝑎𝑛𝑡𝑖𝑙𝑜𝑔 (5.00 − 4.70)
[𝐻𝐴/𝐴] = 𝑎𝑛𝑡𝑖𝑙𝑜𝑔(0.30)
A. Reagents
[𝐻𝐴/𝐴] = 1.995262315
For preparation of Acetate Buffer [𝐵] = 1.995262315[𝐴]
solution, the reagents of Glacial Acetic 2.) Acid-Base Component
Acid and Sodium Acetate are used. For M= A+B
pH analysis, the acid-base indicators are: For Weak Acid Component:
0.50 M = 1.995262315 [A] + [A]
Bromophenol blue, Bromocresol blue,
0.50 M = 2.995262315 [A]
Bromocresol green, Bromocresol purple, [A] = 0.1669302877 mol
Phenol red, Methyl red, Methyl orange, For Conjugate Base Component:
Phenolphthalein and Thymol blue. The B=M-A
assigned sample is Salicylic Acid Solution. B = 0.50 M - 0.1669302877 mol
B = 0.3330697123
3.) Computation for 50ml
B. Procedure For Weak Acid Component:
I. Preparation of Buffers 0.1669302877 𝑚𝑜𝑙/𝐿 = 𝑥/0.50 𝐿
For the preparation of the buffer [A] = .008346514385
solution, the group was assigned to For Conjugate Base Component:
0.3330697123 𝑚𝑜𝑙/𝐿 = 𝑥/0. .050𝐿
prepare a 50 mL 0.50 M Acetate buffer [B] = 0.01665348562
solution with the desired pH = 5.00 and 4.) Computation of Glacial Acetic Acid
pKa = 4.70 and will be tested for its pH. 60𝑔 100𝑔 1𝑚𝐿
. 008346514385 𝑋 𝑋 𝑋
The Weak Acid is Glacial Acetic Acid 1𝑚𝑜𝑙 97𝑔 1.05𝑔
(CH3COOH) and the Conjugate Base is = 0.4916945146 or 0.49 mL Glacial Acetic Acid
(CH3COOH)
Sodium Acetate (C2H3NaO2), in order to
5.) Computation for Sodium Acetate
create Sodium Acetate, NaOH pellets and (NaCH3COOH) components
Glacial Acetic Acid are computed to and [B] = 0.01665348562 x 40g/1 mol
be mixed along with Glacial Acetic Acid to [B] = 0.6661394246 or 0.666g of NaOH
create the Acetate Buffer solution. The [B] = 0.01665348562 x 1/17 mol
[B] = 0.009796168012 or 0.979mL of 17M of
equation used to identify the portions of
CH3COOH
the weak acid and conjugate base
needed for the preparation:
After measuring the components
pH = pKa + log ([A-]/[HA]) of Weak Acid and Conjugate Base and
dissolved together to make a solution, it
[A-] = molar concentration of a conjugate base is dissolved in a small amount (35mL) of
[HA] = molar concentration of an undissociated water. If the desired pH (5.00) was not
weak acid (M)
reached, use the 6.0M HCl or NaOH to for
adjustment of pH. Transferred to a 50mL
Computations:
Volumetric flask, fill to volume and
Given:
labeled properly.
Glacial Acetic Acid (97% w/w, sp.gr.
1.05, MM 60) II. Preparation of Reagents
NaOH pellets (MM 40)
Desired pH: 5.00 In preparing the reagents, 500mL of
pKa: 4.70 6.0M of NaOH and 6.0M HCl is to be used
𝑝𝐻 = 𝑝𝐾𝑎 + 𝑙𝑜𝑔[𝐻𝐴/𝐴] for the adjustment of pHs in the buffer
𝐻𝐴
[ ] = 𝑎𝑛𝑡𝑖𝑙𝑜𝑔(𝑝𝐻 − 𝑝𝐾𝑎) solution, obtaining the desired pH, after
𝐴
being labeled properly and given to
different buffer solutions.

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 prepared Acetate Buffer solution is
not able to reach the desired value, the
addition of reagents of 6.0M HCl or 6.0M
NaOH to achieve the desired pH of 5.00.

IV. Colorimetric Determination of pH

All the needed regeants are placed Figure 1.1 & 1.2.
Application of diff.
in a microplate along with other buffer pH & Acid-Base
solutions in diff. pKa & pH, distilled water Indicators
and assigned sample (Salicylic Acid
solution) in the amount of 200 uL paired
with the different Acid-Base indicators, in III. Results and Discussion
the amount of 20uL. The pH and Concentratiom of
Distilled water, Sample and buffer
The assignment of the microplate
Samples pH [H+]
are:

Distilled Water 7.0 1x10-7

Sample (Salicylic 2.7 1.10x10 -3

Acid)

Buffer Prepared 5.0 1x10 -5

Table 1.2
The table shows the pH (Power of
hydrogen) of the distilled water which is
7.0 and the concentration of hydrogen is
1x10-7. Salicylic acid (sample) with the
pH of 2.7 and ion concentration of
1.10x10-3. Prepared buffer with the pH of
Table 1.1
Assignment of 5.0 and concentration of 1x10-5.
pH and Acid-
Base
Indicator
To compute the concentration of
hydrogen [H+] the formula used is:
[H+]= 10-pH
Colorimetric
Table 1.3
The table shows how the acid-base
indicator reacts with the different pH by
producing colors.
When thymol blue is used as an
indicator to the pH 2.0, 3.0, 5.0, 7.0, 7.5,
8.0, 12.0, distilled water and SA (Salicylic
Acid) the reaction of colors are: salmon
pink, light yellow, light yellow, light
yellow, light yellow,light yellow, light
blue, light yellow, light yellow.
When bromophenol blue is used as
an indicator to the pH 2.0, 3.0, 5.0, 7.0,
7.5, 8.0, 12.0, distilled water and SA
(Salicylic Acid) the reactions of colors
are: yellow, yellow-green, dark blue,
dark blue, dark blue, dark blue, dark
blue, army green and yellow.
When bromocresol green is used as
an indicator to the pH 2.0, 3.0, 5.0, 7.0,
7.5, 8.0, 12.0, distilled water and SA
(Salicylic Acid) the reactions of colors
are: yellow-orange, yellow, blue-green,
blue, blue, blue, blue, blue, yellow.
When bromocresol purple is used as
an indicator to the pH 2.0, 3.0, 5.0, 7.0,
7.5, 8.0, 12.0, distilled water and SA
(Salicylic Acid) the reactions of colors
are: light yellow, light yellow, light
yellow, violet, violet, violet, violet,
grayish green, light yellow.
When methyl red is used as an
indicator to the pH 2.0, 3.0, 5.0, 7.0, 7.5,
8.0, 12.0, distilled water and SA (Salicylic
Acid) the reactions of colors are: red, hot
pink, red, yellow, yellow, yellow, yellow,
red, hot pink.
When methyl orange is used as an
indicator to the pH 2.0, 3.0, 5.0, 7.0, 7.5,
8.0, 12.0, distilled water and SA (Salicylic
Acid) the reactions of colors are: red,
dark orange, orange, yellow-orange,
orange, orange, yellow orange, orange,
red.
When phenolphthalein is used as an
indicator to the pH 2.0, 3.0, 5.0, 7.0, 7.5,
8.0, 12.0, distilled water and SA (Salicylic
Acid) the reactions of colors are:
colorless, colorless, colorless, colorless,
white, light pink, magenta pink, light
pink, white.
When phenol red is used as an
indicator to the pH 2.0, 3.0, 5.0, 7.0, 7.5,
8.0, 12.0, distilled water and SA (Salicylic
Acid) the reactions of colors are: dark
yellow, yellow,yellow, dark yellow, light
orange, light orange, yellow, yellow,
dark-yellow.
Figure 1.3 Reaction of the different pH by
producing colors.
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
(decrease of pH concentration: increase
acidity) or 0.10 M NaOH (increase pH
concentration; increase basicity) 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 is used as a qualitative method
of analysis and determination of the pH
concentration of a chemical substance
through its color intensity. It is 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 is 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.
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
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Scribd:http://www.scribd.com/doc/82682670/
pH-Measurement-and-Buffer-Preparation
● Helmenstine, A. (2019). Henderson-
Hasselbalch Equation. Retrieved
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hasselbalch-equation-and-example-603648W
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