Post-laboratory Report on

Exercise 2
pH and Buffer Systems

Vikki Anne R. Cedo

CHEM 160.1 - 3L
2nd Semester 2014-2015

Groupmates:
Desiree Joy Cerico
Ma. Kriselle Ornales
Mary Ranzelle Pasang

Ms. Korina Vida G. Sinad

Laboratory Instructor

Exercise 2

pH and Buffer Systems

A buffer, as defined by Brown, is a solution that resists drastic pH changes
when limited amounts of acid or base are added to it; an aqueous solution
containing a weak acid and its conjugate base. This weak acid and its conjugate
base are the reason why buffers are able to resist drastic pH changes.
Phosphate buffer system is composed of Na 2HPO4 and NaH2PO4 while the
acetate buffer system is composed of CH3COOH and NaCH3COO.
Upon addition of small amount of acid (H 3O+):
H3O+ + HPO42-

H2O + H2PO4-

H3O+ + CH3COO-

H2O + CH3COOH

Upon the addition of small amount of base (OH -):

OH- + H2PO4-

H2O + H2PO4-

OH- + CH3COO-

H2O + CH3COOH

When H+ is added to the buffer, the weak base component of the buffer
reacts with the H+ and forms the acid component of the buffer. The concentrations
of the acid and base component of the buffer may change some, the ratio of the
[base]/[acid] does not change significantly. This means that the pH will also have no
significant changes. When OH- is added to the buffer, the weak acid is converted to
the base component of the buffer. The [base]/[acid] does not change a lot unless of
course a large quantity of OH - is added, so the pH does not have any significant
change.
It is observed in the experiment that the greater the concentration of the
buffer, the greater is its buffering capacity. In the experiment, different molar
concentrations of phosphate buffer system were prepared from a stock solution of
0.1 M, pH 7.2 The calculations are as follows:
For 0.005 M, V1 =

For 0.5 M, V1 =

For 0.10 M, V1 =

( 0.005 M ) (25 mL)

0.1 M

( 0.5 M ) (25 mL)

0.1 M
( 0.10 M ) (25 mL)
0.1 M

= 1.25 mL of 0.1M phosphate buffer

= 12.5 mL of 0.1M phosphate buffer

= 25 mL of 0.1M phosphate buffer

After the preparation, 2mL NaOH was added to each of the solutions with
different concentrations and the pH of each buffer was recorded in Table 2.1.

As the data in Table 2.1 shows, the buffer with greatest concentration (0.10
M) was the one which resisted drastic pH change, or the most effective buffer. The
ones with lower concentration (0.05 M, 0.5 M) showed significant pH changes
compared to the one with the highest concentration (0.10 M). Theoretically
speaking, the greater the concentration of the weak acid and its conjugate base, the
greater the buffering capacity, i.e., the more strong acid or strong base that can be
neutralized with very little pH change.
Table 2.1. Effect of concentration of buffer on buffering capacity.
Concentration
(M)
Case 1

Case 2

0.005
0.05
0.10

0.005
0.05
0.10

Actual pH
Before addition of
After addition of
NaOH
NaOH
Case 1
Case 2
Case 1
Case 2
7.13
6.98
6.92

4.75
4.54
4.59

12.6
7.41
7.16

13.18
4.97
4.68

pH

Case 1
5.47
0.43
0.24

Case
2
9.03
0.43
0.09

In the addition of a strong base to buffers, the OH - ions react with the best
acid present in the solution (acid component of the buffer HA). This reaction is
assumed to proceed to completion. The OH - rips a proton away from the acid to
produce the conjugate base of the acid and H 2O. Strong bases are great at
accepting protons, that's why it is assumed to proceed to completion. When [HA] =
[A-] (or [BH+] = [B]) for a buffer system, the pH of the solution is equal to the pK a
value for the acid component of the buffer (pH = pK a because [H+] = Ka). The most
effective buffer has equal concentrations of the acid and the base component so it
is equally efficient in absorbing OH - or H+. However, the data in Table 2.2 and 2.3
does not follow the theoretical experiment results. As stated earlier, the most
effective buffer is the one with equal concentrations of its acid and base
component. In other terms, it should be the one with the [A -] / [HA] = 1 or the point
where the buffer's pH = pKa, that will show the smallest pH change but our
experiment results shows that it is the one with [A -] / [HA] = 10. There must be
some errors during the experiment that caused such differences.
Table 2.2. Effect of adding NaOH to buffer with different pH.
Test tube no.

Case
1

1
2
3
4

[A-] / [HA]

0.1
1
10
0.4

Calculated
pH
6.2
7.2
8.2
3.7

Actual pH

pH

Before
addition of
NaOH

After addition
of NaOH

5.73
6.88
7.68
3.11

6.40
7.11
7.72
4.03

0.67
0.23
0.06
0.92

Case
5
6
2
Distilled
water

1
10

4.7
5.7
6.98

3.23
3.43
6.40

4.17
3.90
15.29

0.94
0.47

In addition of strong acids to buffers, the H + always react with the best base
in the solution. The best base has the largest K b and in the buffer system, this will be
the conjugate base (A-) of the acid component of the buffer. The H + reacts with the
conjugate base A- to produce the acid, HA. Strong acids will be donating their proton
to the conjugate base and thus the reaction is also assumed to proceed to
completion. (Bettelheim, 2007).
Table 2.3. Effect of adding HCl to buffer with different pH.
Test tube no.

Case
1

1
2
3
Case
4
5
2
6
Distilled water

[A-] / [HA]

0.1
1
10
0.4
1
10

Calculated
pH
6.2
7.2
8.2
3.7
4.7
5.7
6.98

Actual pH

pH

Before
addition of
HCl

After addition
of HCl

5.73
6.88
7.68
3.11
3.23
3.43
6.40

3.35
6.65
7.31
2.41
2.36
2.31
1.61

-2.38
-0.23
-0.37
0.7
0.93
1.12

When the ratio of the conjugate base to acid is high, there will be a
remarkable change in the pH of the buffer when a strong acid or base is added while
when the ratio of the conjugate base to acid is low, there will be a significant
change in the pH of the buffer. In other words, ratio of the conjugate base to acid is
low, you will observe a relatively high change in the pH of the buffer. This can be
observed in Tables 2.2 and 2.3.

Table 2.4. Titration of unknown amino acid 1.

Volume of 0.1 M KOH
(mL)
0.5
1.0
1.5
2.0
2.5
3.0
3.5

pH
1.53
1.65
1.72
1.80
1.97
2.19
2.47

4.0
4.5
5.0
5.5
6.0

2.79
8.8
9.35
9.55
12.13

Figure 2.1 Titration curve of unknown amino acid 1.

The unknown amino acid 1 is serine, based on the data that we gathered
from the experiment. However, there were some errors during the experiment so
the results weren't the same as the theoretical results. The said errors can be the

wrong handling/ usage of the pH meter (the electrodes were not properly washed
before measuring the pH of the next solution), or excessive / lack of the reagent was
added to the solution. The theoretical pKa1 for serine is 2.21 and the pKa2 is 9.15
and the IpH is 5.68. The amino acid can act as a buffer in its IpH, 5.68. This is
because they are in their zwitterionic form and they react with either a strong acid,
taking a proton or a strong base, by giving a proton.

Figure 2.2 Theoretical titration curve for serine.

Buffers are important in living systems for maintaining homeostasis or
equilibrium inside their bodies to a specific narrow range required to provide
optimum conditions so that biochemical reactions or metabolic reactions may take
place. Even a small change in the pH can be fatal for a living organism. Many
chemical reactions in living cells involve exchanges of hydrogen ions. Because
changes in acidity can affect both the structure and chemical reactivity of cellular
molecules, cells must constantly maintain an acid-base balance.
Buffers are important in biochemical experiments to mimic the pH of the
natural environment where the reactions normally happen and to keep the pH of the
solutions where the reactions are supposed to take place, constant. In the
laboratory, molecular and cellular biologists make extensive use of buffers to
stabilize the pH of aqueous solutions. When studying biomolecules in a test tube,
the biomolecules may be altered or may behave in ways that are uncharacteristic of
their natural behavior if they are in a solution with a pH that is significantly different
from the pH of their natural environment.

References
Bettelheim, F. (2007). Introduction to General, Organic and Biochemistry.
Brooks/Cole by Thomson Learning.
Brown, C. (2005). Introduction to Biochemistry. McGraw-Hill International.
Buffer . (n.d.). Retrieved February 16, 2015, from Encyclopedia Britannica Online:
http://www.britannica.com/EBchecked/topic/83655/buffer
Buffer Solutions. (n.d.). Retrieved February 14, 2015, from Purdue Official Chemistry:
http://www.chem.purdue.edu/gchelp/howtosolveit/Equilibrium/Buffers.htm
Ionization of Amino Acids. (n.d.). Retrieved February 12, 2015, from Chemistry UIUC:
http://butane.chem.uiuc.edu/cyerkes/Chem104ACSpring2009/Genchemref/Ionization
_AA.html
Review in Aqueous Equilibrium. (n.d.). Retrieved February 16, 2015, from Cengage
College:
http://college.cengage.com/chemistry/zumdahl/chemistry/7e/resources/answers_for
_review/ch15_for_review.pdf