pH Measurement and Buffer Preparation

Authors:
David, James Martin Guanzon
Del Rio, Lourdes Antonina
Dela Pedra, Jewel
Dela Rosa, Clarisse
Fontanilla , Rafael
Granil, Hazel
Group 4, 2B-PH, Faculty of Pharmacy, UST

Abstract
This experiment is to allow students in our field of practice to prepare different buffer
solutions, determine the pH of the buffers and samples colorimetrically using different liquid
indicators and electro metrically using the pH meter, and to calculate the buffer capacity of the
prepared buffer solutions. We created buffers of the same kind with different pH levels by using
calculations like the Henderson-Hasselbalch equation. We checked the pH of each buffer by
using two methods; the Electrometric Determination of pH and the Colorimetric Determination
of pH.. The resulting colors for the different indicators showed that the buffer solution has a pH
that ranges from 3-8. Based on our obtained data, it was concluded that the electrometric method
of determining the pH provided a more precise reading compared to the colorimetric method of
determination.

I.

Introduction

The term pH is a convenient way of

expressing low concentration of hydrogen
ions. All biological processes are affected
greatly by pH, which is why pH
determination is very important in our field
of practice especially in developing new
medical products and components. There
are two methods of determining the pH. One
is the electrometric determination. This
method uses a pH meter that involves the
measurements of a cell comprising an

indicator electrode (glass electrode)

responsive to hydrogen ions and a reference
electrode.
Another way of determining the pH is
through the colorimetric method. This was
the method used before the development of
potentiometers and was widely due to their
simplicity. Both methods, however, can only
give approximate values of pH. Colorimetric
determination of pH uses different acid-base
indicators. Indicators are either weak
organic acids or bases. They are substances

that change in color with change in the pH

of the solution they are added. The color
change of the solution with the indicator
depends on the hydrogen ion concentration
of the indicator. Each indicator has a specific
range where visible (color) changes depend
on pH variations. Buffers ,in this
experiment, are tested of its reliability in
calibrating the pH of solutions to near
normal levels (close to pH 7).

II.

Methodology

In order to conduct the experiment,

we prepared Distilled water, Phosphoric
Acid,
Sodium
Hydroxide
pellets,
Hydrochloric Acid, and the acid-base
indicators: Thymol, Bromophenol blue,
Bromocresol green, Bromocresol purple,
Phenol red, Methyl red, Methyl orange, and
Phenolphthalein.
The group prepared a 0.1M
secondary phosphate buffer by mixing 1.7
mL of phosphoric acid (H3PO4), 1.68 g of
sodium hydroxide (NaOH), and enough
distilled water to make 250 mL of the buffer
solution. It was then measured of its pH by
using the pH meter. After recording its initial
pH, it was adjusted to the desired pH which
was 7.5 by slowly adding sodium hydroxide
while being constantly checked with the pH
meter. After doing so, it was transferred to a
suitable container, labeled, and kept for later
use. Acid-base indicators were then
prepared; a drop of each indicator was
placed in its designated test tube and
recorded of its initial appearance. Indicators
used in this experiment are: Thymol,
Bromophenol blue, Bromocresol green,

Bromocresol purple, Phenol red, Methyl red,

Methyl orange, and Phenolphthalein. Two
sets were prepared. The first set was instilled
with 3 mL of water, and the second set was
instilled with the buffer prepared buffer
solution. The color changes were observed
and recorded. The pH range of the buffer
solution and distilled water were then
obtained through the changes of the color of
each sample.

III.

Results and Discussion

The groups buffer preparations

pH was slightly lower than the calculated
pH due to minor experimental errors in
the laboratory. Such errors cannot be
avoided. With that said, we increased its
pH by slowly adding sodium hydroxide
while constantly checking the pH with
the
use
of
a
pH
meter.
The pH meter provided very
accurate pH readings. Quantitative
results were collected such as shifts in
the pH readings during sodium
hydroxide addition due to changes in
hydrogen ion concentration. Based on
the data of the experiment, the
electrometric method is more reliable in
pH
determination.
In the other segment of the
experiment 8 different acid-base indicators
were used to determine the pH of distilled
water and the buffer solution through the
color changes exhibited by the indicators.
The indicators provided a wide range of pH
reading but cannot be as precise as the
electrometric reading. Both tables are

provided at the end of our discussion of

results.
Here are brief descriptions of the
eight indicators.
Thymol: pH range of 1.2 to 2.8 and has a
color change of red (acidic) to yellow
(basic).
Bromophenol: pH range of 3.0 to 4.6 with
yellow (acidic) and purple (basic).
Bromocresol: pH range of 3.8 to 5.4,
yellow (acidic) and blue (basic).
Bromocresol purple: pH range of 5.2 to
6.8, yellow (acidic) and purple (basic).
Phenol Red: pH ranging from 6.4 to 8.2,
with yellow (acidic) and red (basic).
Methyl red: pH ranging from 4.8 to 6.0,
with yellow (basic) and red (acidic).
Methyl orange has a pH that ranges from
3.2 to 4.4, with red (basic) and yellow
(acidic).
Phenolphthalein: pH range of 8.2 to 10.0,
with colorless (basic) and pink (acidic).
Based on gathered data from the experiment,
the method was not precise due to lack of
quantitative result. It merely shows us
visually the reaction of a solution to a
certain pH value.

Samples

Distilled Water
Assigned Samples
(Smart C Juice)
Buffer

HPO4- 0.1
M
Secondary
Phosphate Buffer

pH

8.15

Actual
pH
7.5

[H+]

7.1 x 10

-9

0.00001

[H+]

III.16

x 10
-8

Table.1Electrometric pH Readings

Acid-Base
Indicator

2.0

3.0

7.0

7.5

8.0

12.0

Distilled
Water

Secondar
y
Phosphat
e Buffer
Yellow

Thymol

Red

Yellow

Yellow

Yellow

Yellow

Blue

Red

Bromophenol
Blue
Bromophenol
Green
Bromocresol
Purple
Phenol Red

Yellow

Yellow

Blue

Blue

Blue

Blue

Light
Yellow
Light
Yellow
Yellow

Yellow

Blue

Blue

Blue

Blue

Light
Yellow
Yellow

Violet

Violet

Violet

Violet

Light
Violet
Light
Blue
Yellow

Red

Red

Red

Red

Yellow

Methyl Red

Light
Red
Light
Red
Colorles
s

Light
Red
Red

Yellow

Yellow

Yellow

Red

Yellow

Yellow

Yellow

Orange

Orange

Colorles
s

Colorles
s

Colorles
s

Colorles
s

Yello
w
Yello
w
Red

Light
Blue
Light
Orange
Light
Orange
Yellow

Colorles
s
3-7

Colorless

Methyl Orange
Phenolphthalei
n
pH

Blue

3-8

Table.2Colorimetric pH Determination

IV.

References

[1]
http://chemwiki.ucdavis.edu/Physical_Chemistry/Acids_and_Bases/Buffers/Hende
rson-Hasselbalch_Approximation
[2]
Ferenc Szabadvry (2013-10-22). History of Analytical Chemistry: International
Series of Monographs in Analytical Chemistry. Elsevier Science[3]
[3]
http://wwwchem.csustan.edu/chem1112/1112bufferKS.htm
[4]
Malhotra (2003). Practical Biochemistry for Students. Jaypee Brothers Publishers.
[5]
http://wwwchem.csustan.edu/chem1112/1112bufferKS.htm
[6]
Dr. B. K. Sharma. Analytical Chemistry: (Comprehensively Covering the UGC
Syllabus). Krishna Prakashan Media