Names: Joshua Carvajal Date: February 5, 2020

Clint Danniel N. Guarin

Ginafer S. Enomar

Activity 5: Preparation of Buffers

I. Introduction
A buffer solution contains components that enable the solution to resist large changes in
pH level either acids or bases are added. The may be useful to prepare when aiming to maintain
optimum conditions form chemical reactions. They are in fact routinely used in commercial
products to maintain optimum conditions for product behavior. In our body, blood is a prime
sample of a buffer solution given that it is complex natural buffer solution maintaining a pH level
approximately 7.4, optimum for oxygen transport. Its major buffering agent in blood is the
mixture of carbonic acid and bicarbonate ions.
The action of buffer solutions is the established of an equilibrium between either a b
acid and its conjugate base or a weak base and its conjugate acid. They function in
accordance with the LeChatelier’s principle, which states that an equilibrium system, when
stressed, will shift its equilibrium to relieve stress. For example, adding a large (OH-) to a buffer
solution causes the hydroxide ions to react with H3O+ consumed by the base, maintaining the pH
close to the initial level. Basically the ions of H 3O+ (the stress) is compensated by the
dissociation of acetic acid to produce more H3O+. The process works the same way when an acid
(H3O+) is added to a buffer solution. H3O+ from the acid increases the overall H3O, whom the
system reacts to, to form more molecular acetic acid; the acetate ion combines with H 3O+, so pH
remains close to the initial level. The following experiment will further explore buffer solution.

II. Objectives
At the end of the experiment the researchers must be able to;
A. Identify the acid and salt used in the experiment.
B. Calculate the amount of acid and salt to be used in preparing a buffer solution,
C. Explain the importance of the buffer system in the human body.

III. Materials
Sodium oxalate Beaker
Oxalic acid Graduated cylinder
Distilled water pH paper
2% egg albumin Spatula
Sodium chloride Test tube
Sodium hydroxide Test tube rack
Watch glass
Stirring rod
Dropper
IV. Procedures
A. Preparation of buffer
1. Preparation of buffer solution was given by the instructor. Sodium oxalate for salt and oxalic
acid for acid.
2. Oxalic acid (acid solution) with a concentration of 0.10 M, sodium oxalate (salt solution) with
a concentration of 0.10 M was added into a 200 ml of 0.10 M buffer solution.
3. The volume of the acid and salt as well as the amount of the dissolve chemicals for the buffer
solution was calculated. Graduated cylinder was used to measure the volume and beam balance
to weigh the substance involved in the buffer.
4. The amount of acid and salt to form a buffer solution was calculated.
5. pH level was obtained by using litmus paper.
B. Effect of Buffer
1. 8 test tubes were prepared and were labeled from 1 to 8.
2. Each test tube was added with 5 ml as indicated below;
o Test tube 1 and 2 = distilled water
o Test tube 3 and 4 = 2% egg albumin
o Test tube 5 and 6 = prepared buffer solution
o Test tube 7 and 8 = 0.1 M NaCl
3. Litmus paper was used to get the pH level of the samples.
4. 5 drops of 0.1 M HCl was added to test tubes 1, 3, 5, 7. pH levels were recorded.
5. 5 drops of 0.1 M NaOH was added to test tubes 2, 4, 6, 8. pH levels were recorded.

V. Results and Discussion

A. Data
1. pH and volume of buffer chosen
The pKa values for experiment’s buffer solution range from 1.74 and between 3.81-4.40. The
universal indicator used to calculate its pH indicated a value of 4. The pKa value of 3.81 was used
since it is closer to 4 than 4.40.
Solving for pH, we get:
pH = pKa + log [B-/BH]
pH = 3.81 + log [0.1/0.1]
pH = 3.81
The theoretical pH obtained was 3.81. The buffer solution was prepared with a volume of 200
mL, or 0.2 L.

2. Give the formulas of the acid and salt used to prepared the buffer solution.
Salt: Sodium oxalate (Na2C2O4)
Acid: Oxalic acid (C2H2O4)
 3. Give the reason for the above pair.
The above pair is used since sodium oxalate consists of sodium and oxalate ions in a 2:1 ratio,
and is an oxalate salt and an organic sodium salt. Basically, sodium oxalate is the sodium salt
of oxalic acid with the formula Na2C2O4.

4. Calculate the volume of the acid and salt.

Acid (Oxalic Acid): C2H4O4 (0.02 mol) (26.07 g/mol) = 1.8 g
Salt (Sodium Oxalate): N2C2O4 (0.02 mol) (1.34 g/mol) = 2.68 g

5. Experiment pH
Using the pH paper: pH value of 4

6. Is the experiment pH greater than or equal to the desired pH?

The theoretical pH was 3.81, so the experimental pH is greater.

7. If the experimental pH obtained is greater than the desired pH, what substance (acid or salt) is
adjust the experimental pH to the desired pH?
Since the experimental pH is greater than the desired pH, an acid would be needed to
decrease it until it matches the desired pH of 3.81.

B. Effect of buffer
Table A. The table below shows the pH changes before and after putting acid HCl and the base
NaOH.
3

Test Tube no. pH before the pH after pH before the pH after pH after the
addition of addition of addition of addition of addition of
0.1 M HCl HCl 0.1 M NaOH 0.1 M NaOH HCl and
NaOH
1. 5 1
2. 5 12
3. 6 4
4. 6 14
5. 3 3
6. 3 4
7. 5 1
8. 5 13

a. Calculations:
1. Solution 0.5 M Hac and 0.5 M NaCH*COO (mixed) makes a buffer solution. What will happen
to the buffer solution if 0.1 M NaOH is added?
Similar to the results from the experiment above, the buffer solution will be resistant to
pH changes even after the 0.1 M NaOH. Of course, the amount of change is dependent on the
volume of the buffer solution that is needed to be produced and how much NaOH is added
 to the solution. In the experiment, the salt (sodium oxalate) and acid (oxalic acid) had the same
molarity and were mixed together to form the buffer solution. After a few drops of NaOH, the
value of the pH only increased by 1 (from 3-4). The same should also be true for this particular
problem, given that they have similar premises.

2. Prepare a buffer solution with the pH OH 9 using NH3 and NH4Cl. (pKa of NH3 = 9.25)
a. If the concentration NH3 is 0.5 M. What is the concentration of the salt?
pH = pKa + log [B-/HB]
pH - pKa = log [B-/HB]
10 –(pH-pKa) = [B-/HB]
[HB] = [B-]/10-(pH-pKa) = 0.5/10-(9-9.25) = 0.281 M

3. Calculation for the volume of acid and salt in preparing a buffer solution for the experiment.
Acid (Oxalic Acid): C2H4O4 (0.02 mol) (26.07 g/mol) = 1.8 g
Salt (Sodium Oxalate): N2C2O4 (0.02 mol) (1.34 g/mol) = 2.68 g

VI. Questions with Answers

1. What is the significance of buffer in living organism?
Biological fluids in our body are buffered to maintain the pH of our circulatory and
digestive fluids in a specific range, such that the associated process respond to a given acidity.
Clearly, homeostasis is assisted by buffering, through the maintenance of pH around a specific
value.

2. Summarize the preparation of buffer.

The most common equation when it comes to buffer solutions is the Henderson-
Hasselbach approximation, which is particularly useful when calculating for equilibrium
concentrations. The equation comes from
HA + H2 ↔ H3O + A-
Where A- is the conjugate base and HA is the weak acid. The buffer process is described by an
equilibrium constant expression. For acids, Ka is used to represent acids, with the subscript a
implying an acid equilibrium. Using a few mathematical maneuvers, we eventually get
pKa = pH – log [A-]
where pKa is -log [Ka]. From here, the pH can be solved with the following equation:
pH = pKa + log [A-]
where [A-] is the conjugate base over the acid in moles. Obtained results from this equation
will be the pH of the buffer solution after the reaction has taken place. We can also solve for the
necessary amounts of salt and acids in order to produce a buffer solution with a specific pH.

3. With respect to the pH changes, redude and discuss the result of the test tube 1 and 2, 3 and 4,
5 and 6, 7 and 8.

Test Tube no. pH before the pH after pH before the pH after

 addition of 0.1 addition of addition of 0.1 addition of 0.1
M HCl HCl M NaOH M NaOH
1. 5 1
2. 5 12
3. 6 4
4. 6 14
5. 3 3
6. 3 4
7. 5 1
8. 5 13

As can be seen from the table, test tubes 1 and 2, which contain distilled water, show pH
changes depending if an acid (HCl) or base (NaOH) is added. The same goes for test tubes 3 and
4 (contains egg albumin) and test tubes 7 and 8 (contains NaCl). If HCl is added, their pH
values decrease. If NaOH is added, their pH values increase. However, for test tubes 5 and 6,
which contain the buffer solution, hardly showed any significant pH change. Adding the HCl did
not change its pH value of 3, and the addition of NaOH only slightly increased to 4. According to
the definition of a buffer solution -- a solution whose components enable it to resist large
changes in pH level either acids or bases are added – it makes sense that the buffer solution
would resist changes in its pH after the addition of HCl and NaOH.

4. Based on the experiment, what can you say about the buffer and pH changes?
The buffer solution hardly changed its pH despite adding an acid or a base, which makes
sense because a buffer solution is supposed to function that way, that it would resist changes in
its pH value. In other words, it functions to maintain pH close to a specified value.

VII. Conclusion
As mentioned earlier, in our body, the blood pH maintenance is controlled through the
bicarbonate buffer. This system consists of ions such as carbonic acid and bicarbonate. This
buffer acts to form carbon dioxide gas when blood pH drops into acidic range. During the
respiration process, the lungs expel this gas out of the body. This buffer brings pH back to
neutral during alkaline conditions by causing excretion of the bicarbonate ions through the urine.
Besides blood, there are other biological buffers. The phosphate buffer system acts similarly to
the bicarbonate buffer, but it does have much stronger action. This buffer comprises hydrogen
phosphate ions and dihydrogen phosphate ions, in the internal environment of all cells. It reacts
with the hydrogen phosphate ions, which accept them, under conditions where excess hydrogen
enters the cell. The dihydrogen phosphate ions accept the excess hydroxide ions entering the cell,
under alkaline conditions. Proteins consist of amino acids held together by peptide bonds. The
amino acids possess an amino group and a carboxylic acid group. At physiological pH, the
carboxylic acid exists as the carboxylate ion (COO-) with a negative charge and the amino group
exists as the NH3+ ion. When the pH becomes acidic, excess hydrogen ions are taken up by the
carboxyl group to return to the form of carboxylic acid. If the pH of the blood becomes alkaline,
the NH3 + ion releases a proton that takes the form of NH2. Hemoglobin, the respiratory
pigment present in the blood, also has buffering action within tissues. It has the ability to bind at
 a given moment with either protons or oxygen. Binding one sets the other off. In hemoglobin,
binding of protons occurs in the portion of globine while binding of oxygen occurs at the portion
of heme iron. At the time of exercise, protons are generated in excess. Hemoglobin helps in the
buffering action by binding these protons, and simultaneously releasing molecular oxygen.

VIII. Documentation

Before adding HCl and NaOH: pH levels

Prepared Buffer Solution

NaCl pH level
pH level

Distilled water pH level 2% egg albumin pH level

After adding HCl and NaOH: pH levels

Distilled water added Distilled water added
with HCl with NaOH

2% egg albumin added 2% egg albumin added

with HCl with NaOH

Prepared buffer solution Prepared buffer solution

added with HCl added with NaOH
 0.1 M NaCl added with 0.1 M NaCl added with
HCl NaOH

IX. References
Retrieved from “General, Organic, and Biochemistry” by Denniston, Topping, and Caret
Retrieved from https://sciencing.com/acidity-levels-functional-groups-5389525.html
Retrieved from https://socratic.org/questions/why-are-buffers-important-to-living-things
Retrieved from https://chem.libretexts.org/