MODULE 2: PH AND BUFFER SOLUTIONS B.
PROPERTIES OF ACIDS AND BASES
MLS2E - GROUP 4
CABASAN, Edward Theodore
KANAAN, Anne Therese
PARIÑAL, Christian Haj
PASCUAL, Rengel
I. INTRODUCTION
Acids and bases can be defined according to
their compositions and molecular structures. At
present, the Bronsted-Lowry Theory, proposed by
Johannes Bronsted and Thomas Lowry, is
considered the most useful definition of acids and
bases. To further apply this theory, this module is
geared towards determining the importance of the
pH and buffer systems in biological processes.
In this laboratory experiment, the Medical
Laboratory Science (MLS) students mastered the
properties and measurements of pH in varying
solutions. The students were able to determine how
buffer systems operate on a macroscopic scale.
From these concepts, they were able to calculate
and determine the pH of different solutions using a
laboratory simulator, litmus solutions, and utilized a
buffer capacity equation.
II. METHODOLOGY
A. DETERMINATION OF PH VALUE USING
LAB SIMULATOR
Six different solutions were chosen
from the drop-down menu of the lab
simulator: drain cleaner, hand soap, water,
coffee, orange juice, and battery acid. A
beaker was then filled with 500 mL of each of
the six chosen solutions. A probe was
dragged to the first solution to measure the
pH value using a pH meter. The same
procedure was used for the succeeding
chosen solutions.
a.) Reactions of Hydrochloric acid (HCl),
and Sodium hydroxide (NaOH) to Blue
and Red Litmus Solutions
Two test tubes were positioned in a
test tube rack. Red litmus solution was
poured in test tube A, and blue litmus
solution in test tube B. A few drops of
Hydrochloric acid (HCl) were extracted and
added to both the red litmus solution and
blue litmus solution. The MLS students
observed the change in color of each
solution. The same procedure was followed
for Sodium hydroxide (NaOH).
b.) Reactions of Hydrochloric acid (HCl),
and Sodium hydroxide (NaOH) to Zinc
Metal
A minute amount of Zinc metal was
taken using a spatula and was transferred to
a clean test tube. A few drops of Hydrochloric
acid (HCl) were poured to the same test tube
and was clamped to the stand. A single-
bored cork was utilized to close the test tube.
A jet tube was inserted through the test tube
and with a Bunsen burner, the test tube was
heated. At the same time, a match was lit and
brought near the mouth of the test tube. The
students observed the flame on the match.
The same procedure was followed for
Sodium hydroxide (NaOH).
c.) Reaction of Hydrochloric acid (HCl) to
Solid Sodium Carbonate
The neck of a flat-bottom flask was
clamped and placed in a stand. A minute
amount of solid Sodium carbonate was
added in the flask followed by distilled water.
The flask was then covered with a double-
bored cork. A thistle funnel was inserted
through one of the holes in the cork. A beaker
containing lime water was positioned near
the flat-bottom flask. Next, one end of a
connecting tube was inserted into the flask
through one of the holes in the cork, ensuring
Page 1 of 6
 that the tube was inserted securely and was
Water 7 Neutral
not touching the solution in the flask. The
other end of the same connecting tube was Coffee 5 Acid
inserted into the beaker containing lime
water. Then, a small amount of Hydrochloric Orange Juice 3.50 Acid
acid (HCl) was poured into the flask through
a thistle funnel. The students observed the Battery Acid 1 Acid
reaction.

B. PROPERTIES OF ACIDS AND BASES

d.) Reaction of Sodium hydroxide to Solid
Sodium Carbonate
Table 2. Determination of properties of acids and
bases
A minute amount of Sodium
carbonate was transferred into the test tube
using a spatula. After which, a small amount Solution Reaction Interpretation
of Sodium hydroxide (NaOH) solution was
also poured, and the test tube was shaken HCI in Color of the HCI is an Acidic
gently until the solid Sodium carbonate Blue Solution Solution
dissolved completely. A few drops of Litmus changed
Phenolphthalein were poured to the test Solution from Blue to
tube. The students observed the reaction. Red

NaOH in Color of the NaOH is a Basic

C. BUFFERS AND BUFFER CAPACITY Red Solution Solution
Litmus changed
To determine the buffer capacity, first, a pH Solution from Red to
meter was used to measure the initial pH value of Blue
250 mL Hydrochloric (HCl) solution and 250 mL
buffer solution (Acetic acid and Sodium acetate HCI + Zn Produced Acids react with
solution). Next, a few drops of methyl red indicator metal Zinc metals
solution were added on each beaker and transferred chloride
10 drops of 0.1 M Sodium hydroxide solution to each and
beaker. The pH value for each solution was Hydrogen
measured again. After measuring the pH value for Gas
each solution, the corresponding buffer capacity was
NaOH + Produced Basic solutions react
calculated by dividing the moles of hydroxide
Zn metal Sodium with metals
present in the solution over the change of pH value.
zincate and
Hydrogen
gas
III. RESULTS
HCI + CO2 is There is a reaction
Na2CO3 produced between acidic and
A. DETERMINATION OF PH VALUE USING which turns alkaline solutions
LAB SIMULATOR Lime water
milky
Table1. Determination of pH Value using a Lab
Simulator NaOH + Solution There is no reaction
Na2CO3 changed to between two similarly
Solution pH value Interpretation pink basic or alkaline
solutions;
Drain Cleaner 13 Basic Phenolphthalein
produce the pink color
Hand Soap 10 Basic
Page 2 of 6
 C. BUFFER AND BUFFER CAPACITY to blue from red indicates it is a base. A neutral
litmus solution is usually purple.

Table 3. Determination of buffer capacity In the conducted experiment, the litmus

solution changed from blue to red confirming the
pHi pHf pH Interpretatio acidity of Hydrochloric acid (HCl). Meanwhile,
Solutio Differenc n Sodium hydroxide (NaOH) turned the red litmus
n e solution to blue, confirming its alkalinity.

HCl 4.7 9.8 5.19 As a non- Acids and bases react with metals to produce
0 9 buffer salts and Hydrogen gas. This is evident in the
solution, separate reactions of HCl and NaOH with Zinc in the
results experiment. The reaction of HCl with Zinc produced
showed a Zinc chloride and Hydrogen gas while the reaction of
significant NaOH with Zinc produced Sodium zincate and
change in pH Hydrogen gas. The “Squeaky Pop” test was used to
confirm the evacuation of Hydrogen gas from the
Acetic 4.7 4.7 0.03 As a buffer opening of the jet tube. The test was confirmed when
Acid 0 3 solution, an audible pop was recognized after the matchstick
results was placed into the opening of the tube. During this
showed a test, a single displacement reaction occurred in
resistance in which Zinc replaced the Hydrogen in the two
the change in solutions. The two reactions can be summarized by
pH the following equations:

2Zn+ 2HCl ⇋ ZnCl2 + H2

IV. DISCUSSIONS
Zn + 2NaOH ⇋ Na2ZNO2 + H

A. DETERMINATION OF PH VALUE
When solid Sodium carbonate was mixed
with HCl, it produced Carbonic acid and Sodium
The pH value of a solution varies depending
chloride, or more colloquially known as table salt.
on the presence of the hydrogen and hydroxyl ions.
Carbonic acid is known to be an unstable compound.
Based on the experiment, when water was added to
Carbon dioxide through brisk effervescence passed
a weak acid or base, the solution became more
through the tube and mixed with lime water which
basic. More specifically, the addition of a solvent
formed a Calcium Carbonate precipitate thereby
such as water to a given solution results in an
resulting in a milky appearance in lime water. The
increase in pH for acidic solutions whereas the pH
double displacement reaction can be summarized
otherwise decreases with the addition of water in
by the following equation:
basic solutions as the action reduces the
concentration of ions in the solution. As such, the
amount of a solution does not affect its pH value but
Na2CO3 + 2HCl ⇋ 2NaCl + H2CO3
only its concentration.

B. PROPERTIES OF ACIDS AND BASES
The use of litmus solutions is one of the most
widely used methods in determining the alkalinity or
acidity of a solution. The solution changes color
when it comes into contact with the solution. A
changing litmus solution to red from blue often
indicates that the solution is an acid while changing
When Sodium hydroxide and solid Sodium
carbonate were mixed in the test tube, no visible
reactions were observed because both were alkaline
solutions. This was confirmed by the change in color
from transparent to pink due to the exposure of
Phenolphthalein in the alkaline medium.

Page 3 of 6
 C. BUFFER AND BUFFER CAPACITY

Buffers are created using a weak acid and a

soluble ionic salt of the acid or a weak base and a
soluble ionic salt of the base. Buffers can resist
drastic changes in pH by neutralizing any solution
added to it. However, all buffers are only effective
within their specific area of buffering capacity which
stresses the importance of having the knowledge to
use the right buffers in certain pH ranges. Buffers are
highly essential solutions not only in Chemistry but
also in other science-related endeavors as they are
used to keep biological specimens under the right
pH for accurate testing and experimentation.

V. REFERENCES

Libretexts. (2020, August 22). pH Indicators.

Chemistry LibreTexts.
https://chem.libretexts.org/Bookshelves/Physical_a
nd_Theoretical_Chemistry_Textbook_Maps/Supple
mental_Modules_(Physical_and_Theoretical_Chem
istry)/Acids_and_Bases/Acid_and_Base_Indicators/
PH_Indicators

Helmenstine, Anne Marie, Ph. D. (2020, August 28).

What is a Universal Indicator in Chemistry Retrieved
from https://www.thoughtco.com/definition-of-
universal-indicator-605761

American Chemical Society. (2016). pH and Color

Change | Chapter 6: Chemical Change | Middle
School Chemistry. 2020 American Chemical
Society. https://www.middle
schoolchemistry.com/lessonplans/chapter6/lesson8

Helmenstine, Anne Marie, Ph.D. (2020, August 28).

pH Definition and Equation in Chemistry. Retrieved
from https://www.thoughtco.com/definition-of-ph-in-
chemistry-604605

Boyer, R., 2012. Biochemistry Laboratory Modern

Theory And Techniques. 2nd ed. Prentice Hall.

Page 4 of 6
POST-LAB QUESTIONS:
1) Discuss how the following buffer systems
work in the human body:
a) Carbonate Buffer System
Also termed as the Carbonic Acid Buffer
System, the Carbonate Buffer System aids in
maintaining the flow of blood in the human body.
The system works by regulating the pH in the
bloodstream within its 7.35-7.45 range, as it
depends on the ratio of carbon dioxide to
bicarbonate. It uses carbon dioxide and water to
create a weak acid called Carbonic acid that
dissociates to one hydrogen ion and
Bicarbonate. This reaction is reversible and
hence, in the same light, Carbonic acid can also
be created by the addition of one hydrogen ion
to the Bicarbonate. Through this process,
potential acid and base imbalances are
managed and regulated by the said buffer
system by controlling and keeping a constant
pH in the bloodstream. As such, this mechanism
supports proper metabolic functions in the body.
The greater one inhales, the greater
amount of carbon dioxide is exhaled and while
the lungs are at work while breathing, the
kidneys help the pulmonary organ maintain
homeostasis by excreting acids and bases into
the blood. A well-regulated pH in the
bloodstream deviates from the occurrence of
acidosis and alkalosis. To summarize the entire
flow of the Carbonate Buffer System, the
following equation is utilized:
CO2 + H2O ↔ H2CO3 ↔ H+ + HCO3- .
Sources:
https://sciencing.com/carbonate-buffering-
8299150.html
https://socratic.org/questions/what-are-the-
three-major-buffer-systems-of-the-body-
and-how-do-they-work
b) Phosphate Buffer System
While the Carbonate Buffer System
functions within the bloodstream, the Phosphate
Buffer System works within the cell’s internal
fluids. This buffer system is composed of
phosphate groups that operate in regulating the
pH levels of fluids inside the human body. The
inorganic phosphate groups present in the
system freely bind to free hydrogen ions to act
as a buffer. More specifically, the dihydrogen
phosphate ions in the buffer system function as
hydrogen ion donors, or acids in general, while
the hydrogen phosphate ions work as hydrogen
acceptors, or the bases. The dihydrogen
phosphate ions dissociate to hydrogen and
hydrogen phosphate to allow the decrease in pH
and promote the formation of dihydrogen
phosphate from hydrogen, and hydrogen
phosphate to promote increase in pH. This
reaction is reversible and can be further
summarized through the equation:
H2PO4- ↔ H+ + HPO42-.
The buffer system uses H2PO4- and
HPO42-. The pK of inorganic phosphate is at 6.8
which encompasses the pH of extracellular fluid
at 7.4 with a low concentration in its intracellular
space. This makes the Phosphate Buffer
System critical in the renal tubes and
intracellular fluids of the human body as its
concentration continues to increase as the
amount of fluids absorbed in the tubules
likewise increases. As a whole, this interaction
among phosphates maintains the overall pH
level within and around the cells ensuring that
the body maintains its required level of acidity.
Sources:
https://socratic.org/questions/what-are-the-
three-major-buffer-systems-of-the-body-
and-how-do-
theywork#:~:text=Phosphate%20buffer%20
system%20operates%20in,the%20ion%20a
cceptor%20(%20base%20)%20.&text=Prot
ein%20buffer%20system%20helps%20to%
20maintain%20acidity%20in%20and%20aro
und%20the%20cells.
Page 5 of 6
 c) Proteins
To further maintain acidity within and
among cells, proteins perform within the body’s
intracellular fluids and blood plasma. Since
proteins are strongly made up of amino acids,
their compositions are built towards the
presence of positively charged amino groups
and negatively charged carboxyl groups
wherein the charged molecules interact by
binding to hydrogen and hydroxyl ions and
hence function as buffers in the human body.
work#:~:text=hydrogen%20phosphate%20io
n.,PROTEIN%20BUFFER%20SYSTEM,are
%20also%20good%20at%20buffering.
https://quizlet.com/144447324/chapter-27-
flash-cards/
https://open.oregonstate.education/aandp/c
hapter/26-4-acid-base-
balance/#:~:text=Hemoglobin%20as%20a%
20Buffer,by%20the%20dissociation%20of%
20oxygen.

Proteins such as albumin mainly https://melcalin.com/albumin-actions-and-

function as pH buffers in blood plasma by interactions-benefits-from-a-buffered-
reducing the chances of coagulation to occur in albumin-supplementation-pralbumina/
the blood as well as in healing processes. Due What Is a Protein Buffer System?
to the presence of histidine residues in albumin, (healthfully.com)
acid dissociation among fluids is made possible
and albumin becomes an effective buffer by Phosphate Buffer System - Medical
donating positive charges during alkalosis or Physiology - Euroform Healthcare
negative charges during acidosis.
pH Buffers in the Blood (wustl.edu)
Another protein buffer system that works
in the human body is hemoglobin, which permits
the red color in blood. Hemoglobin contains 2) A sodium hydroxide solution has a pH of
alpha and beta protein chains that have a heme 11.40, what is the [OH–] in this solution? Show
group containing iron. Due to this heme group, your solution
oxygen can bind to the iron atoms in hemoglobin
and be transported through the blood. With this,
hemoglobin functions as a buffer system by
binding to minute amounts of acids in the blood
while stabilizing any potential rises of pH in the
bloodstream by releasing hydrogen ions from the
atomic sites present within the hemoglobin
molecule. Likewise, to restabilize a decrease in
the pH, hydrogen ions are taken up or engulfed
by hemoglobin to obtain homeostasis.

3). An HCl solution has a pH of 3.10, what is the

Sources: [H3O+] in this solution? Show your solution
https://socratic.org/questions/what-are-the-
three-major-buffer-systems-of-the-body-
and-how-do-they-

Page 6 of 6