Name: Coloso, Ma. Jinelle V.

Date Performed: February 16,2016

Section: 3ChE-A Group No. 5 Date Submitted: April 5, 2016

Experiment No. 5
Spectrophotometric Determination of pKa

INTRODUCTION

Almost all of the known compounds in the chemical industry contain different acid or
basic groups that could determine ones physical and chemical properties. The different acid or
basic groups can be determined through spectrophotometry. Spectrophotometry is a method
wherein the device, a spectrophotometer, measures how much a chemical substance absorbs or
transmits light. The device can also measure the amount of chemical substance present in a
particular material. In this experiment a UV-VIS spectrophotometer was used [1].

Another important matter in the experiment is the determination of pKa. pKa measures the
strength of an acid solution depending on the chemical properties of the acid. The smaller the
pKa, the more acidic is the substance. In aqueous solution, the equilibrium of the pKa can be
written as [2]:

HA + H2O A- + H3O+ equation (1)

Where HA and A- represents the acid and base indicator, respectively. The chemical
substances is said to be in equilibrium if their concentrations do not change through time. The
determination of Ka can be calculated using the formula:

[ ][ + ]
HA A- + H+ : Ka = equation (2)
[]

Getting the logarithm of equation (2), the equation will be in the form:

pK = pH log equation (3)

Wherein may be determined through the use of a spectrophotometer.
 Two wavelengths will be chosen (maxima and the minima) which will be the basis of the
two equations:

A1 = a1,HA [HA] + a1,A [A-] equation (4)

A2 = a2,HA [HA] + a2,A [A-] equation (5)

The experiment was focused on the determination of the equilibrium constant of the
dissociation of an indicator in water using spectroscopic method. The experiment was also
focused on the demonstration of the principle of equilibrium and the interaction of matter with
light.
METHODOLOGY

Fig. 5.1 The Hitachi UV-Visible

Spectrophotomoter used.

Fig 5.2 The prepared solutions from 1 to 10 that was used in the experiment

0.1 M NaH2PO4 was prepared by mixing 0.3011g of NaH2PO4 in a 25mL of water. The
same method was used for the preparation of 0.1M Na2HPO4. 0.355g of Na2HPO4 wax mixed in a
25mL of water. Lastly 0.1M of NaOH was prepared by adding 0.1782g NaOH to 25mL of water.
(See Appendix A for the calculations). Then, 10 solutions were made which covers the entire pH
range of 1-13. A solution of 2 to 9 was prepared by mixing the right amount of 0.1M NaH2PO4
and 0.1M Na2HPO4 and was diluted to 25 mL as indicated in the table. (See the Laboratory
Manual in Physical Chemistry for Engineers 2 page 33 for the table). Afterwards the pH for
solutions 2 to 9 was adjusted using the pH meter by adding 0.1M HCl to alkaline solutions and
0.1M NaOH to acidic solutions. The adjusting of pH was done to make the solutions pH equally
distributed in the range of 1-13. (See Table 5.1 for the final pH of the solutions). Afterwards,
solution 1 was prepared by adding 1.0mL of methyl red indicator to 4.0mL of distilled water and
0.2mL of concentrated HCl and was afterwards diluted to 25mL. For solution 10, 1.0mL of
methyl red indicator was added to 24.0mL of 0.1M NaOH. Then the cuvette was rinsed thrice
with the solutions being measured. Before reading, the absorbance of solution 1 and 10 was set
from 350nm to 800 nm with a 10nm intervals in between. Also, the difference of both solutions
was taken into consideration. Through this, the maxima and minima of the curve were obtained.
The data was recorded. Lastly, the maxima and minima of the curve of solutions 1 and 10 were
used to measure the absorbance of the other 8 solutions with the use of the photometric method.
(See table 5.2). (For the pictures of Experiment 5, See Appendix B)

RESULTS AND DISCUSSIONS

Ten (10) solutions were used in this experiment. Furthermore, 4 points were discussed
for the treatment of results. The following results were obtained:
Table 5.1 pH of the 10 solutions
Solution No. pH Adjusted pH
1 2.22 2.22
2 4.76 5.97
3 6.38 6.38
4 6.85 6.80
5 7.20 7.20
6 7.48 7.60
7 7.89 8.04
8 8.30 8.44
9 9.17 8.81
10 12.86 12.86

After plotting the absorbance of solutions 1 and 10 and getting the difference between
the two, the students obtained a 0.9520 at 530nm as its maxima and -1.5290 at 420nm as its
minima.
 Wavelength Vs. Absorbance
Absorbance Solution 1 Absorbance Solution 10 Soln 1 - Soln 10

1.5

0.5

0
0 100 200 300 400 500 600 700 800 900
-0.5

-1

-1.5

-2

Figure 5.3. The Wavelength vs. Absorbance of Solutions 1 and 10.

The maxima and minima were used to determined the absorbance of the remaining
solutions using the photometric method.
Table 6.2 Absorbance of Solutions 2 to 9
Solution No. 530nm 420nm
(Maxima) (Minima)
2 0.121 0.400
3 0.078 0.598
4 0.047 0.495
5 0.089 1.458
6 0.079 1.614
7 0.053 1.480
8 0.030 0.734
9 0.060 1.500

Using equations (4) and (5) and the absorbance at its minima and maxima, the a-values
were calculated wherein the concentration of HA is 0.1M and A- is 0 when pH is 1 and the
concentration of HA is 0 and A- is 0.1 when pH is 13.

0.996=1, HA (0.1)+ 1, A- equation (6)

0.049=2, HA (2,) (0.1)+ 2, A- equation (7)
0.044=1, HA (1,) (0)+ 1, A- (0.1) equation (8)
1.578=2, HA (2,) (0)+ 2, A- (0.1) equation (9)
 Solving simultaneously equations (6) to (9), the students obtain
1, HA = 9.96 2, HA = 0.49
1, A- = 0.44 2, A- = 15.78

Afterwards, the students obtained the values of [HA] and [A] using again equations (4)
and (5) and the calculated values of 1, HA, 1, A-, 2, HA, and 2, A-. Finally the log ratio of [A] and
[HA] was determined and was plotted versus the pH. Equation (3) was used for the
determination of the log ratio.
Table 6.3 Determination of the Ratio log A/ HA
Solution No. pH as adjusted *[HA] *[A-] pK log A / HA

1 2.22 0.1 0 - -
2 5.97 1.10E-02 2.50E-02 5.615 0.355
3 6.38 6.17E-03 3.77E-02 5.594 0.786
4 6.80 3.34E-03 3.13E-01 4.828 1.972
5 7.20 4.86E-03 9.22E-02 5.922 1.278
6 7.60 3.42E-03 1.02E-01 6.124 1.476
7 8.04 1.80E-03 9.38E-02 6.322 1.718
8 8.44 9.59E-04 4.65E-02 6.754 1.686
9 8.81 1.83E-03 9.50E-02 7.094 1.716
10 12.86 0 0.1 - -
-
(See Appendix A for the calculations of [HA] and [A ])

log In / HIn vs. pH

2.5

1.5

0.5

0
0 2 4 6 8 10 12 14

Fig. 5.4 The log A-/ HA vs. pH of solutions 1 to 10.

 Averaging the calculated pKs of solutions 1 to 10, the students obtained a value of
6.032. Comparing the calculated value from its literature value which is 5.05, the students
obtained a 19.45% error. The error came from different causes like the adjusting of pH wherein
from solutions 2 to 9 the pH was distributed equally but not for solutions 1 and 10 which may
have contributed an error to the experimental value of pK. Reading of the cuvettes may also be
the cause for the cuvette was used several times which may have contaminated the next readings.
ANSWERS TO QUESTION

The particular wavelengths of a certain substance can determine how a certain person
could perceive the object, thus, is responsible for determining the different colors a person can
deduce. Another point is the analysis of colors is carried mostly in its maximum point wherein
the absorption coefficient is least sensitive to changes[3].

pKa contributed a lot in the determination of a substance if it is a weak acid or a strong

acid. A smaller pKa value indicates the substance to be acidic. Another important use of the pKa
is its relation with the pH value. Once the two are in equilibrium, half of the acid had already
dissociated [4].

The indicator of a substance may vary depending if the environment or substance to be

tested is in basic, or acidic. For example in a titration of a weak acid with a strong base a good
indicator would be the phenolphthalein for it changes color in a basic pH [5].

CONCLUSION AND RECOMMENDATION

Using spectrophotometric method, one can determine the pKa of a certain substance.
Once the minima and maxima of the curve were determined, the absorbances for the other
solutions are attainable. With this, HA and A- may be calculated by solving simultaneously
equations (4) and (5). Averaging, the students obtained a value of 6.032 which has a percent
error of 19.44%. The theoretical value of the dye is 5.05 at room temperature. Some factors have
contributed to the error. One was when the pH is being adjusted wherein from solutions 2 to 9
the pH was distributed equally but not for solutions 1 and 10 which may have contributed an
error to the experimental value of pK of the dye. Reading of the cuvettes may also be the cause
for the cuvette was used several times which may have contaminated the next readings. The
temperature had also contributed for the theoretical value is only exclusive for room
temperatures. The students performed the experiment in a much cooler environment. Lastly,
when the pH was being adjusted using the pH meter, the slope that was obtained was only 91%
which ensures a 9% inconsistency to the true reading.

Some recommendations are: if through the use of buffers, the pH meter could not be set
at 100%, a linear regression must be used. Another is that the cuvette must be rinse thoroughly
for every reading for an undesired substance will contaminate the desired reading. Lastly,
checked the environment if the conditions for the theoretical value is the same.
APPENDIX A

PREPARATION OF Na2H2PO4, NaOH and Na2HPO4:

Weight of Na2H2PO4 (in grams):

0.1M = x ; x = 0.0025 moles
0.025L
0.0025 moles x 119.98g/mole Na2H2PO4 = 0.3g Na2H2PO4
Weight of Na2HPO4 (in grams):
0.1M = x ; x = 0.0025 moles
0.025L
0.0025 moles x 141.96g/mole Na2H2PO4 = 0.35g Na2H2PO4
Weight of NaOH (in grams):
0.1M = x ; x = 0.0025 moles
0.025L
0.0025 moles x 40g/mole Na2H2PO4 = 0.1g Na2H2PO4

CALCULATIONS FOR [HA] AND [A-]:

Solution 2:
0.121= (9.96)[A] + (0.44)[A-] [HA] = .
0.400= (0.49)[A]+(15.78) [A-] [A- ] = .
Solution 3:
0.078= (9.96)[A]+(0.44) [A-] [HA] = .
0.598= (0.49)[A]+(15.78) [A-] [A-] = .
Solution 4:
0.047= (9.96)[A]+(0.44) [A-] [HA] = .
0.495= (0.49)[A]+(15.78) [A-] [A-] = .
Solution 5:
0.089= (9.96)[A]+(0.44) [A-] [HA] = .
1.458= (0.49)[A]+(15.78) [A-] [A-] = .
Solution 6:
0.079= (9.96)[A]+(0.44) [A-] [HA] = .
1.614= (0.49)[A]+(15.78) [A-] [A-] = .
Solution 7:
0.053= (9.96)[A]+(0.44) [A-] [HA] = .
1.480= (0.49)[A]+(15.78) [A-] [A-] = .
 Solution 8:
0.030= (9.96)[A]+(0.44) [A-] [HA] = .
0.734= (0.49)[A]+(15.78) [A-] [A-] = .

Solution 9:
0.060= (9.96)[]+(0.44) [A-] [HA] = .
1.500= (0.49)[]+(15.78) [A-] [A-] = .
APPENDIX B

Fig 5.5: 0.3550g of Na2HPO4 Fig 5.6: 0.1782g of NaOH

Fig 5.7: 0.3011g of NaH2PO4

REFERENCES

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https://en.wikipedia.org/wiki/Acid_dissociation_constant

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