NAMES: Gabriel, Esther Faith/ Garma, Czarina/ Javier, Jonah/ Herrera, Lorlyn/ Ibea, Hardee

DATE: November 27, 2014

Experiment No. 1
BUFFERED SYSTEMS
I. ABSTRACT
Buffers are aqueous systems that resist changes in pH when small amounts of acid or
base are added. Buffer solutions are composed of a weak acid (the proton donor) and its
conjugate base (the proton acceptor). Buffering results from two reversible reaction equilibria in
a solution wherein the concentration of proton donor and its conjugate proton acceptor are
equal. For example, in a buffer system when the concentration of acetic acid and acetate ions is
equal, addition of small amounts of acid or base does not have any detectable influence on the
pH.
There are several factors that could affect the pH of the buffer. First is the accuracy of its
preparation. One should be careful of the proper weighing and pipetting of the substances
needed to prepare the buffer solution. Second is the addition of Sodium chloride which basically
decreased the pH of the buffer. This is due to increase in the ionic strength of the solution. Third
is the temperature which has relative effect on the pH depending on the buffer used. Fourth is
the addition of water to the buffer. Theres an increase of pH because there is a reduction of
ionic strength.
Buffers also play an important role in our biochemical systems in the maintenance of pH
within narrow limits. We can see in the three important buffer systems in our bodies:
1. bicarbonate buffer system
2. phosphate buffer system
3. protein buffer system
Therefore buffers are very important in our cells because all of our enzymes need a specific
pH value to work at optimum. If the pH changes our enzymes might deteriorate which means
death.
II. OBJECTIVES:
At the end of the experiment, the student will know:
1. What buffers are and how are they prepared.
2. The mechanism of buffering.
3. The relevance of buffers in the human body.
4. Some buffer systems in the body.
III. MATERIALS:
A. Glassware and Apparatus used:
1. Pipet and aspirator

2. Weighing balance
3. Vials
4. Beaker
5. Graduated cylinder
6. Volumetric Flask
7. Plastic Bottles
B. Chemicals used and its properties:
Structure
1. Acetic Acid

2. Sodium
Acetate

3. NaH2PO4

4. Na2HPO4

Properties/Uses
Acetic acid is a monocarboxylic acid because it
contains only one COOH group. It has a sour
taste and pungent smell. It is the main component
of vinegar. Vinegar is typically 3-7% solution of
acetic acid in water. Vinegar is mainly used as a
preservative in food and in the pickling of
vegetables. Water free acetic acid is known as
glacial acetic acid.
Sodium acetate is the sodium salt of acetic acid. It
has the chemical formula C2H3O2Na and is also
known as sodium ethanoate. It is an inexpensive
chemical that has a wide range of uses, including
as a food additive and pickling agent or a
laboratory reagent. It is also the prime ingredient in
portable, reusable, chemical-based heating packs.
Monosodium phosphate (NaH2PO4), also known
as anhydrous monobasic sodium phosphate and
sodium dihydrogen phosphate, is an inorganic
compound of sodium with
dihydrogen phosphate (H2PO4-) anion. One of
many sodium phosphates, it is a common industrial
chemical. It exists as an anhydrous salt, as well as
mono- and dihydrates.
Disodium hydrogen phosphate is used as an in
conjunction with trisodium phosphate in foods and
water treatment. In foods, it is used to adjust pH.
Its presence prevents coagulation in the
preparation of condensed milk. Similarly, it is used
as an anti-caking additive in powdered products.
The pH of disodium hydrogen phosphate water
solution is between 8.0 and 11.0, meaning it is
moderately basic

5. NaCl

Sodium chloride is readily soluble in water and

insoluble or only slightly soluble in most other
liquids. It forms small, transparent, colorless to
white cubic crystals. Sodium chloride is odorless
but has a characteristic taste. It is an ionic
compound, being made up of equal numbers of
positively charged sodium and negatively charged
chloride ions. When it is melted or dissolved in
water the ions can move about freely, so that
dissolved or molten sodium chloride is a conductor
of electricity; it can be decomposed into sodium
and chlorine by passing an electrical current
through it.

6. NaOH

Sodium hydroxide is a white deliquescent solid

having melting point at 591 K (318C). It is stable
towards heat. It is highly soluble in water and
considerable amount of heat is evolved due to the
formation of a number of hydrates e.g., NaOH.H2O,
NaOH.2H2O.

IV. DISCUSSION: Data and Results

Table 1. pH measurements of the four buffers
pH
Calculated
3.6
4.0
7.6
8.0

Buffer 1
Buffer 2
Buffer 3
Buffer 4

Electrometric
4.4
5.6
7.8
8.6

a. Provide theoretical discussions for your results.

The calculated and electrometric pH of the four buffers did not match. This may be
because of the inaccurate weighing and pipetting of the reagents. Another is that, while
pipetting the solution it might have been contaminated and thus it affected the pH of the
buffers.
Table 2. Effect of temperature and salt on buffers

Buffer 1
Buffer 2

pH with NaCl
3.5

Diluted pH value
5.5

pH/C
5.8

Buffer 3
Buffer 4

6.9

8.3
8.9

a. Give explanation on your results.

With the addition of 0.1 ml of 1M NaCl, the pH of buffers 1 and 3 decreased. NaCl
is neutral in nature but with its addition it increases the ionic strength of the solution.
Ionic strength affects ionization constants, activity of ionized species (salt form) of the
buffer, and activity of hydronium ion which alters pH of solution.
On the other hand, addition of water (dilution) to a buffer solution can alter its pH
due to reduction in ionic strength. The following buffer equation for an acid (HA) and its
conjugate base (A-) accounts for the ionic strength by incorporating activity coefficient
term, A- :

activity coefficient is usually less than 1. Accordingly, log A- is negative. Thus, it is

expected to have lower pH for acid- conjugate base pair when ionic strength is
accounted for.
Temperature change affects Ka, Kb, and Kw. Therefore, pH of buffers is influenced
by temperature. Each buffer is influenced differently. Some buffers decrease in pH (e.g.
boric acid- sodium borate) while others increase in pH (e.g. acetic acid- sodium acetate)
as temperature increase. Moreover, the magnitude of change is relatively small for acidic
buffers when compared to basic buffers.
b. How would the sodium chloride affect the ionic strength of buffer 1 and buffer 3?
The setting of the ionic strength of a buffer solution should be done in the same
way as the setting of the pH value when selecting the electrolyte, since this increases
depending on the electrolyte used. The multivalent ions contribute strongly to the ionic
strength. Ionic strength is equal to the concentration.
Table 3. Buffer capacity
3 ml
6 ml
9 ml
12 ml
15 ml

pH
9.4
11.5
13.9
16.1
18.3

Buffer Capacity
0.09
0.07
0.06
0.06
0.05

a. Is the buffer capacity a fixed value for the given buffer system? Justify your answer based on
your tabulated results.
No. It depends on the amount of base or acid added.
b. What is the effect of increasing concentration of NaOH as increments of the base added?
With the addition of more NaOH, the buffer capacity decreases rapidly.

c. When does a buffer have its greatest buffer capacity?

The buffer has its greatest capacity before any base is added where [salt] / [acid] =
1, and according to equation, pH = pKa.
QUESTIONS:
1. What is the structure of protein that can account for their buffering capacity? Answer briefly
and completely.
Amino acids are carbonic compound that contain two functional groups. One is
the amino group (NH2) and a carboxyl group (COOH). Protein structure are the bonds by
which amino acid chains generate complex protein structure.
2. A buffer contains 0.05 mole/liter of formic acid and 0.1 mole/liter of sodium formate. The pKa
of formic acid (HCOOH) is 3.75. What is its pH?

3. What is the ratio of bicarbonate to carbonic acid in normal blood with a pH of 7.4? (pKa
H2CO3 = 6.36)
20:1

V. CONCLUSION
Upon the completion of the experiment we learned that buffers are solutions that resist
the change in pH when small amounts of an acid or base is added. It is made of a weak acid
and its conjugate base. A weak acid and its conjugate base can remain in solution without
neutralizing each other.

When hydrogen ions are added to a buffer, they will be neutralized by the base in the
buffer. Hydroxide ions will be neutralized by the acid. These neutralization reactions will not
have much effect on the overall pH of the buffer solution. This is how buffer works.
Buffers are important in our body. Reactions inside the human body take place in the
blood plasma. These reactions might fail to happen if the pH changes. For complete reaction to
take place, the pH of the blood should remain constant. Biochemical reactions are quite
sensitive to the nature of blood. The reaction inverts by changes in the pH of blood. However,
these buffers generally prevent such mishap. These changes also affect the biological activity of
a human being.
There are several buffer systems in our body but probably the most important is the
bicarbonate buffer system. The pH of human blood must be maintained between ranges of 7.07.8. For example, when you drink a soda, its acid neutralizes the small of base in your blood.
The remaining acid in the soda causes the pH of your blood to decrease rapidly to a pH of about
5. Because the enzymes in your body don't work well at a pH of 5, you die a horrible death. But
thanks to our blood which is a very effective buffer, it can stabilize our bodys pH.

VI. REFERENCES
Charmaine, M. (2011). Importance of buffer solution. Retrieved November 24, 2014 from
http://www.ehow.com/about_6637932_importance-buffer-solution.html.
Helmenstine, A.M. (2014). What are buffers and what do they do? Retrieved November 24,
2014 from http://chemistry.about.com/od/acidsbase1/a/buffers.htm.
Gharaibeh, S. (2013). Factors affecting pH of buffer solutions. Retrieved November 24, 2014
from http://phcalculator.blogspot.com/2013/06/factors-affecting-ph-of-buffersolutions.html.

Mohan, C. Buffers: A guide for the preparation and use of buffers in biological systems. San
Diego, California: Calbiochem