Basic Experiments

in Physical Chemistry

2022-1

Prof. : 한귀영 교수님

Student ID:
Name:
Assignment name: Lab #7 Result report
Date: 2022.04.13

Sungkyunkwan University
School of Chemical Engineering
 [Lab 7] Dissociation Constant of Weak Acid

1. Result
1) Data sheet

pH 3 pH 4 pH 5 pH 6 pH 7 pH 8 pH 9 pH 10 pH 11
0.038 0.043 0.057 0.366 1.819 2.296 3.780 3.208 3.532
0.041 0.048 0.060 0.388 2.117 2.486 3.673 3.616 3.463
0.037 0.044 0.058 0.393 2.089 2.602 2.954 3.232 3.430
0.047 0.040 0.059 0.396 2.062 2.973 3.060 3.648 3.684
0.039 0.044 0.058 0.390 2.125 2.726 3.535 3.032 3.688
0.038 0.040 0.059 0.387 2.128 3.448 2.898 3.731 3.425
Table 1. Experimental data sheet for the absorbance in different pH condition.

Among the measurement data obtained using the spectrophotometer, the values with errors

are indicated with lines. (Data displayed in red letters on the screen of the device)

pH 3 pH 4 pH 5 pH 6 pH 7 pH 8 pH 9 pH 10 pH 11
0.038 0.043 0.057 N/A N/A 2.296 N/A 3.208 3.532
0.041 N/A N/A 0.388 2.117 2.486 N/A N/A 3.463
0.037 0.044 0.058 0.393 2.089 2.602 2.954 3.232 3.430
N/A 0.040 0.059 0.396 2.062 2.973 3.060 N/A 3.684
0.039 0.044 0.058 0.390 2.125 2.726 N/A 3.032 N/A
0.038 0.040 0.059 0.387 2.128 N/A 2.898 N/A 3.425
Average
0.0386 0.0422 0.0582 0.3908 2.1042 2.6166 2.9706 3.1573 3.5068
Table 2. Data sheet in which the average value is calculated after removing one or more
abnormal data (abnormal data is indicated as N/A)
Among the measured data obtained using the spectrophotometer in Table 1, the values with
errors (data displayed in red letters on the screen of the device) were also excluded from the
average value calculation (indicated in red N/A).

2) Graph and its analysis using least square method

Fig1. It is a graph showing the average absorbance at each end point connected by a curve.
Mean absorbance versus pH and horizontal asymptotes were plotted.

Absorbance lies in the middle of two asymptotes with appropriate mathematical properties. In
this case, linear regression should be used to find the pH value. If the X-axis is PH and the Y-
axis is Absorbance, the linear regression equation has the form of a linear function. (Y = AX
+ B)
It can be expressed using the least squares method as in the following equation.

Using the least squares method shown above, the results are presented in a table as follows.

Table 3. Values calculated using least squares method.

Using this method, Y can be expressed as a linear function with respect to X.

Y = 0.521148X – 1.99412
The median absorbance of the two asymptotes is (3.5068+0.0368)/2 = 1.7718, so
When the absorbance at this point is 1.7718, the pH can be calculated.
X = (1.9456+1.99412)/0.521148 = 7.5597  pH = 7.5597

3) Comparison of experimental values and theoretical pKa values of BTB solution

The experimentally calculated pH is 7.5597 at the intermediate absorbance of the two
asymptotes.
At this time, when BTB is ionized by the HInd ↔ H+ + Ind- reaction, the concentrations of
HInd and Ind- are the same. You can then use the Henderson-Haselbach equation, a formula
that can be used in buffers.

log( [Ind] / [Hlnd] ) is zero. So pKa is equal to pH = 7.5597 in the experiment.

Theoretically, the pKa value of BTB is 7.1. Therefore, we can compare the pK values of the
experimental and theoretical values and then calculate the error rate between them.
 Error rate: (7.5597-7.1)/7.1 ×100%=6.47465%

4) Comparison of [Ind-] and [HInd] at each pH state

Using the Henderson-Hasselbach equation

log ([Ind])/[Hlnd]) can describe the magnitude relationship between [Ind-] and [HInd] at each
pH state.

2. Discussion
 1) Possible errors
i) generation of air bubbles
If bubbles are generated when the BTB solution is added to the buffer solution, it causes an
error in measuring the volume. In addition, bubbles cause errors because they can cause light
refraction and reflection when measuring data using a spectrophotometer.
ii) mixing of solutions
In the experiment, the solution was mixed well by pipetting. However, just visually
confirming that the color is uniform does not guarantee that the solution is completely mixed.
A solution that is not completely mixed will cause errors in the measurement with a
spectrophotometer.
iii) Average calculation of 6 absorbance measurements
Accurate experimental values can be obtained as the number of samples increases and the
number of replicates increases in all experiments. In addition, the number of samples was
insufficient because the data whose values differed too much from other data among the six
data were excluded. This will cause errors.
iv) Difference between linear regression and actual value
Linear regression was used to calculate the pH value at the midpoint of the two asymptotes.
However, the calculated values obtained using linear regression may not exactly match the
actual values.

2) How to minimize errors

i) When adding each solution to the 96 plate using a pipette, be careful not to create
bubbles in the solution.
ii) More pipetting to thoroughly mix the solution can reduce the error. Another way
is to transfer to a tube after it is completely mixed, rather than visually checking
that the color is evenly spread.
iii) increase the number of data samples; Six pieces of data were obtained for each
pH, and the error can be reduced by calculating with much more data than six.
The error can also be reduced by repeating the experiment several times and
comparing the calculated values in each experiment. The sample is smaller
because the data with errors on the data display screen of the spectrophotometer
(data displayed in red text on the screen) were removed and calculations were
made.
iv) In order to reduce the error between the value calculated by the linear regression
method and the actual value, the error can be reduced by further subdividing the
X-axis interval of the linear regression. For example, if the pH is tested not at 3.0,
4.0, ... 11.0, but at more detailed pH intervals such as 3.0, 3.5, 4.0, 4.5, ..., the
error will be reduced.
3. Conclusion
1) The goal of the experiment
Through this experiment, the dissociation constant of the BTB solution can be obtained
through absorbance analysis by understanding the process of equilibration of weak acids
under different pH conditions and analyzing it using a spectrophotometer.

2) Experimental contents and error analysis

i) Experimental contents
Analysis with a spectrophotometer allows to obtain experimental data on absorbance at
various pH conditions. After removing outliers from this data, an absorbance graph
according to pH can be drawn by calculating the average value.
After drawing the graph, the pH at the absorbance corresponding to the middle of the two
asymptotes can be calculated. (using linear regression)
The calculated pH value by substituting in the Henderson-Haselbach equation, the pKa
value can be found and compared with the theoretical value, and the magnitude
relationship between [Ind-] and [HInd] can be revealed at each pH state.
ii) Analysis of errors
In order to reduce the error, bubbles should not be generated in the process of transferring
the solution with a pipette, and if there are many bubbles, the experiment process should
be repeated. For better mixing, more pipetting should be done to reduce the error.
In addition, in order to reduce the error by increasing the sample size of the data, more
solutions can be used or the number of repetitions of the experiment can be increased. If
the experiment is conducted by further subdividing the pH range, the error between the
calculated value and the theoretical value can be further reduced.

3) Lessons learned through experiments

Through this experiment, you can understand the concepts of pH and pKa and the
relationship between ion concentration used in the Henderson-Hasselbach equation. You
will also learn how to calculate data obtained using a spectrophotometer using least
squares and linear regression.

4. Answers for the result report

1) Question 1: It is in 1. 1)
2) Question 2: It is in 1. 1)
3) Question 3: It is in 1. 2)
4) Question 4: It is in 1. 2)
5) Question 5: It is in 1. 3)
6) Question 6: It is in 1. 4)

5. References
Sungkyunkwan University School of Chemical Engineering – (2020) Chemical
Engineering & Polymer Basic Experiments, p39-45.
Chemlibertexts-https://chem.libretexts.org/Ancillary_Materials/Reference/
Organic_Chemistry_Glossary/Henderson-Hasselbach_Equation
Ibm- https://www.ibm.com/topics/linear-regression