Finalpapersophomoreyear

The Kinetics of Crystal Violet Fading
Patrick Ducusin, Nafshiyat Nihal and Ethan Price
Macomb Mathematics Science Technology Center
Honors Chemistry
Mrs. Hilliard, Mr. Supal and Mrs. Dewey
23 May 2018
Ducusin – Nihal – Price
Table of Contents
Introduction………………………………………………………………………………………1
Review of Literature...…………………………………………………………………….…….3
Problem Statement……………………………………………………………………………...8
Experimental Design……………………………………………………………………………9
Data and Observations………………………………………………………………………..11
Data Analysis and Interpretation……………………………………………………………..26
Conclusion......................................................................................................................36
Work Cited......................................................................................................................39
1
Introduction
One may believe crystal violet to be a simple and useless but beautiful dye.
However, the dye can be used for much more than some think. Crystal violet is a bright
purple organic compound which fades to a colorless dye as it is mixed with solutions of
high pH. One of the crystal violet dye’s most important applications is in the Gram Stain
method which helps to classify bacteria.
The absorbance of crystal violet in a crystal violet and sodium hydroxide solution
was measured over time to determine the order of the reaction with respect to crystal
violet and the rate constant. Also, the absorbance of an alternative solution that had a
lower sodium hydroxide concentration was measured over time to determine the order
of the reaction with respect to sodium hydroxide. All of this was needed to find out the
order of the overall reaction. The order of the reaction is important because it gives
information as to how the rate of a reaction changes based on the concentration of the
reactants. Furthermore, this research served as a pathway into the complexity of
chemical kinetics and how different factors could affect the rate of a reaction.
Absorbance was measured because tools like spectrophotometers and
colorimeters can only measure absorbance accurately and not concentration of a
reactant. The absorbance was then converted into concentration through Beer
calibration curve that was found before trials, and this was necessary in order to
graphically analyze the concentration of crystal violet over a period of time. From there,
2
the order with respect to crystal violet could be determined based on the characteristics
of the graph as well as other key components of the rate law like the rate constant.
This research is important as it provides insight into the factors that affect the
rates of chemical reactions. Scientists can use similar methods that were used in this
paper to obtain or analyze data but with other types of items and chemical reactions.
They can look into how temperature affects the deterioration of certain vegetables, how
certain chemicals in the air increase the decomposition rate of the ozone layer, or what
can increase the yield of products in certain types of reaction.
Crystal violet and sodium hydroxide were the reactants available for this
experiment because when they react, the solution fades and becomes almost colorless.
The kinetics of the color fading reaction could easily be analyzed by measuring the
absorbance with a colorimeter so data collection would not be too tedious, and a Beer-
Lambert calibration curve was used to easily convert absorbance to concentration. Also,
because the concentration of the sodium hydroxide was made to be greater than the
concentration of crystal violet, a simpler rate law could be used that would not require
more experiments of alternating solutions. However, an alternative solution was made
anyways to determine the order of the reaction with respect to sodium hydroxide even
though it was not needed for the rate law.
Rate laws appear everywhere in the world, either through industrial, biological, or
other means. Almost every living organism in the world has chemical reactions
happening within their bodies, and they cannot be too drastic in change or else it could
endanger the organism’s life. Also, in chemical industries, it is important that the most of
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product is yielded everyday. It works on other scales like how fast medicine reacts or
how fast food can spoil. Reactions and rate laws play an important role in everyday life.
Review of Literature
Kinetics is the field which is concerned with the rates, or speeds, of reactions,
and it is one of the most important subjects in chemistry. It can relate to how fast
medicine is able to work in for certain types of diseases as well industrial problems such
as the developments of catalysts to synthesize new materials. Also, when dealing with
the rates of reactions, many different factors can affect the rate of reactions due to the
different properties of substances.
C25N3H30Cl (crystal violet) is a bright purple dye which, and due to its structure as
a monovalent cation, it fades into a colorless solution when mixed with solutions of high
pH . Crystal violet is a triphenylmethane dye; also known as an intensely colored
organic compound. This dye is an important compound. Crystal violet is used in Gram’s
Method which is a method of classifying bacteria (Gram Stain).
In chemical reactions, there are rate laws. These make connections with the rate
of a reaction based on other variables. In the reaction with crystal violet, the rate of the
reaction changes as the concentration of the dye and hydroxide ions vary. Rate laws of
chemical reactions can be determined by observing the concentration changes over
time. This can be accomplished by using either spectroscopy or colorimetry (Knutson).
4
Figure 1. The Electromagnetic Spectrum
Spectroscopy is the study of the interaction between matter and electromagnetic
radiation. Specifically, an object’s light is split into its independent colors. The color
pertains to wavelength on the visible light spectrum. When there is higher energy in
waves, the wavelengths are shorter. On the contrary, lower energy waves have longer
wavelengths (Kulesa).
Figure 2. The Mechanism of a Spectrophotometer
A spectrophotometer is the instrument that will be used. This tool compares the
initial amount of light that passes through a material with the final amount of light that is
absorbed by a certain sample. A spectrophotometer can be used to distinguish
compounds since different compounds absorb different wavelengths, hence, their
different colors. Also, Beer’s Law states that the amount of light that is absorbed is
5
directly proportional to the concentration of the chemical compounds that are absorbing
the light (Michigan State University). This allows for measuring concentrations of crystal
violet in the experiment and comparing the amount of light absorbed by it which will help
to determine the rate law for the dye.
The reaction rate of a chemical reaction is the speed at which a chemical
reaction occurs. However, the rate is represented in a specific way. This is represented
as the change in concentration of a solution over time. The reaction rate (M/s) is how
the molarity (M which is found by dividing number of moles by liters) of a solution
changes over a period of time (seconds). The change in the concentration is found by
subtracting the initial concentration of the reactants from the final concentration while
the rate is found by dividing the change in molarity by the change in time (final time
minus initial time) and flipping the sign to make it positive. The sign is flipped because
the final concentration of the reactants minus the initial concentration gives a negative
number, but the reaction rate is represented as a positive number (California State
University, Sacramento).
In graphs of concentration v. time, curves are steeper at the beginning of a
reaction and flatten out over time. This means that higher concentrations of reactants
increase the rate of the reaction because there are more opportunities for collision
between molecules. Because of this relationship, chemists express the rates in terms of
the concentration of reactants in which rate = k[A]x[B]y. The sum of X and Y is the order
for the entire reaction, and reaction order is the number that expresses how the rate
changes when the concentration changes, and it is to respect with both reactants A and
6
B in a reaction, and it is important to note that the overall reaction order determines the
units of the rate constant.
The collision of molecules in a chemical reaction are affected by temperature,
orientation, and catalysts. Chemical reactions occur when molecules collide with
enough chemical energy to break chemical bonds. This is called activation energy
which is represented by Eₐ. As temperature increases, the kinetic energy of a molecule
also increases. Since not all of the molecules in the substance have sufficient energy to
react, increasing the temperature only increases the fraction of the molecules that can
engage in chemical reactions. Another factor that affects collision is orientation. The
orientation of a molecule affects collision because when the reactants and products
collide in certain orientations, an activation complex (combination of reactants) is
formed which splits into the products. Catalysts, on the other hand, provide another way
for the chemical reaction to take place which takes less activation energy (Norton
StudySpace).
When determining reaction rates, the Arrhenius equation relates activation
energy, the rate constant, and the effects of temperature and orientation on the rate.
The rates of chemical reactions increase with temperature because a greater amount of
molecules hold enough kinetic energy to overcome activation energy. When put
together, the Arrhenius equations states that:

−𝐸𝑎
𝑘 = 𝐴𝑒 ( 𝑅𝑇 )
Figure 3. The Arrhenius Equation
7
Figure 3 is the entire equation in which A is equal to the frequency factor, e is
Euler’s number, Ea is the activation energy in joules, R is the gas constant 8.314
J/k*mol, and T is the temperature in Kelvin. The frequency factor expresses the
importance of orientation of the molecules during collisions because the greater the A
value, the more likely collisions will result in a chemical reaction.
Several experiments were found that relate to this experiment being conducted.
One of which measures the absorbance of hydrogen peroxide in quantities of crystal
violet as well as in peroxidase. In the experiment, crystal violet and peroxidase is mixed
into hydrogen peroxide resulting in a bluish or purplish mixture. The absorbance of the
mixture is measured against a mixture of the same composition minus hydrogen
peroxide (Mottola). This experiment is similar to our experiment in the fact that it
measures the absorbance of a mixture composed of a substance and crystal violet.
Another experiment was found that was similar to our experiment. In the
experiment, crystal violet was mixed with hydroxide ions. An empirical crossover model
was used to analyze the UV spectrophotometry of the crystal violet and hydroxide ions
(Du). This experiment relates to ours because it uses spectrophotometry to analyze a
crystal violet mixture.
8
Problem Statement
Problem:
The purpose of this experiment was to determine the rate law of the color-fading
reaction between crystal violet and hydroxide ions. The experiment provided the
information of how rate laws worked as well as its relevance in chemical kinetics.
Hypothesis:
When the concentrations of the aqueous solutions of crystal violet and sodium
hydroxide double, the color fading reaction will be a second-order reaction.
Data Measured:
In the experiment, the independent variable was the volume of 1% crystal violet
solution (ml). The dependent variable was the rate of the chemical reaction of crystal
violet. This change depends on what order the reaction is (how the reaction rate
changes based on the concentration). If it is first-order, the unit is s⁻¹. The units are
M⁻¹s⁻¹ for second order and M⁻²s⁻¹ for third-order reactions. The control variable in the
experiment was the volume of distilled water (500 ml) for the solution as well as the
9
volume for diluting the solution which was also 500 ml. Throughout the experiment, the
absorbance (AU) for the crystal violet and NaOH was measured. Since the absorbance
and concentration are proportional to each other, the molarity (M which is found by
moles/liter) was found next for both which allowed for monitoring the molarity over time
to find the rate of the reaction. The statistical analysis for this experiment was
descriptive statistics to analyze the different initial reaction rates for each solution. This
lead to finding the actual order of the reaction for the color-fading reaction.
Experimental Design
Materials:
25 µM Crystal violet, (2.5 x 10-5), 50 mL Pipet bulb

Sodium hydroxide solution, NaOH, 0.02 Pipet, serological, 10-mL
M, 500 mL Colorimeter
Water, distilled or deionized Stirring rod
Beaker, 50 mL Stopwatch or timer
Cuvettes or test tubes Verneir logger pro
Kimwipes or lens tissue
Safety Precautions:
Dilute sodium hydroxide solution is irritating to eyes and skin. The crystal violet stock
solution is flammable. Crystal violet is a strong dye and will stain clothes and skin.
Clean up all spills immediately. Wear chemical splash goggles, chemical-resistant
gloves, and a chemical-resistant apron. Avoid contact of all chemicals with eyes and
skin. Be sure to neutralize the crystal violet and hydroxide solution with acid and plenty
of water to dispose it down the drain.
Procedures for Calibrating the Colorimeter:

*Turn on the colorimeter and let it warm up 20 minutes before use.
1. Calibrate the machine by setting the colorimeter to 565 nm using the arrows.
2. Make a “blank solution” of equal volumes of distilled or deionized water and 0.02
M NaOH. Place the solution into a cuvette.
10
3. Wipe the sides of the cuvette with the kim wipes and insert it into the sample
compartment. Remember to line up the clear side of the cuvette with the arrow in
the sample compartment when placing it in.
4. Close the sample compartment press the CAL button to calibrate the machine.
5. After the red light stops blinking, take out the blank solution from the sample
compartment.
Procedures for Taking Readings:
1. Measure 10.0 mL of 25 μM crystal violet in a serological pipet and add it to a

clean 50-mL beaker.
2. Rinse the pipette with distilled water several times and also with the sodium
hydroxide solution. Measure 10.0 mL of 0.02 M sodium hydroxide in a serological
pipet.
3. Add the sodium hydroxide into the 50-mL beaker with crystal violet.
4. Mix and immediately press “begin timing”.
5. Transfer the reacting solutions to a cuvette and clean the outside with a lint-free
wipe.
6. Place the cuvette into the colorimeter and close the lid. Record absorbance
measurements every 20 seconds for 15 minutes.
7. Take the solution out of the colorimeter
8. Measure 10.0 mL of 25 μM crystal violet in a serological pipet and add it to a

clean 50-mL beaker.
9. Rinse the pipette with distilled water several times and also with the sodium
hydroxide solution. Measure 5.0 mL of the sodium hydroxide solution with the
serological pipet.
10. Add the sodium hydroxide into another 50-mL beaker and add in 5.0 mL of
distilled water to reduce the molarity of the sodium hydroxide solution to 0.01 M.
11 Take the sodium hydroxide solution (0.01 M) made in step 10 and add it into the
beaker with the crystal violet.
12. Mix and immediately press “begin timing”
1
13. Transfer the reacting solutions to a cuvette and clean the outside with a lint-free
wipe.
14. Place into the colorimeter and close the lid. Record absorbance measurements
every 20 seconds for 15 minutes.
15. Take the solution out of the colorimeter.
16. Repeat steps 1-15 about 5 times to complete 10 trials. Always calibrate the
machine first before taking measurements.
Diagram:
Sample Compartment Lid
2
Mode button
Wavelength
Wavelength
Sample Compartment
Modes
Arrows and CAL

button
Cuvette
Figure 4. Mechanism of the Colorimeter
The figure above shows the parts of the colorimeter that are relevant for
calibrating the machine.
Data and Observations

3
Data:
Below are the tables for both the 0.02 M and 0.01 M concentration solutions as
well as the observations for each trial.
Table 1
Absorbance of Crystal Violet for Trial 1
Time(s.) Absorbance(Au) Time(s.) Absorbance

0 N/A 460 0.095
20 N/A 480 0.091
40 0.184 500 0.089
60 0.177 520 0.087
80 0.167 540 0.084
100 0.163 560 0.082
120 0.157 580 0.079
140 0.153 600 0.077
160 0.149 620 0.074
180 0.144 640 0.072
200 0.139 660 0.069
220 0.135 680 0.067
240 0.131 700 0.065
260 0.127 720 0.064
280 0.122 740 0.062
300 0.120 760 0.060
320 0.116 780 0.058
340 0.112 800 0.057
360 0.109 820 0.055
380 0.107 840 0.054
400 0.104 860 0.052
420 0.100 880 0.050
440 0.098 900 0.049
Table 1 shows the absorbance of the crystal violet and sodium hydroxide solution
for 15 minutes.
Table 2
4

0 N/A 460 0.095
20 N/A 480 0.092
40 0.184 500 0.089
60 0.178 520 0.087
80 0.173 540 0.085
100 0.168 560 0.083
120 0.162 580 0.080
140 0.157 600 0.078
160 0.153 620 0.076
180 0.148 640 0.074
200 0.143 660 0.072
220 0.139 680 0.069
240 0.135 700 0.067
260 0.130 720 0.064
280 0.126 740 0.062
300 0.123 760 0.061
320 0.119 780 0.059
340 0.115 800 0.058
360 0.112 820 0.055
380 0.109 840 0.054
400 0.106 860 0.052
420 0.101 880 0.051
440 0.098 900 0.049
for 15 minutes.
Table 3
5

0 N/A 460 0.142
20 N/A 480 0.140
40 0.189 500 0.138
60 0.186 520 0.136
80 0.183 540 0.134
100 0.181 560 0.132
120 0.179 580 0.131
140 0.176 600 0.129
160 0.174 620 0.128
180 0.172 640 0.127
200 0.169 660 0.125
220 0.167 680 0.124
240 0.165 700 0.122
260 0.163 720 0.121
280 0.161 740 0.120
300 0.159 760 0.119
320 0.157 780 0.117
340 0.154 800 0.116
360 0.152 820 0.114
380 0.150 840 0.113
400 0.148 860 0.112
420 0.146 880 0.110
440 0.144 900 0.109
for 15 minutes. In this trial, the molarity of the sodium hydroxide solution was 0.01 M
because it was important to see how the rate of the reaction changed when the
concentration of the reactant was changed. The absorbance also never went below
0.100.
Table 4
6

0 N/A 460 0.151
20 N/A 480 0.149
40 0.194 500 0.146
60 0.191 520 0.143
80 0.189 540 0.142
100 0.187 560 0.141
120 0.184 580 0.150
140 0.182 600 0.149
160 0.180 620 0.147
180 0.178 640 0.145
200 0.176 660 0.144
220 0.174 680 0.142
240 0.171 700 0.140
260 0.169 720 0.138
280 0.167 740 0.137
300 0.166 760 0.135
320 0.164 780 0.133
340 0.162 800 0.132
360 0.160 820 0.131
380 0.158 840 0.129
400 0.156 860 0.128
420 0.154 880 0.127
440 0.152 900 0.125
for 15 minutes. Like in Trial 3, the molarity of the sodium hydroxide was 0.01 M and the
absorbance never went below 0.100
Table 5
7

0 N/A 460 0.110
20 N/A 480 0.106
40 0.198 500 0.103
60 0.191 520 0.100
80 0.187 540 0.097
100 0.181 560 0.094
120 0.176 580 0.091
140 0.171 600 0.089
160 0.167 620 0.087
180 0.162 640 0.084
200 0.158 660 0.083
220 0.153 680 0.080
240 0.149 700 0.079
260 0.145 720 0.077
280 0.141 740 0.075
300 0.137 760 0.074
320 0.133 780 0.072
340 0.130 800 0.071
360 0.126 820 0.069
380 0.123 840 0.067
400 0.119 860 0.067
420 0.116 880 0.065
440 0.113 900 0.064
for 15 minutes. In this trial, the molarity of the sodium hydroxide solution was 0.02 M.
Table 6
8

0 N/A 460 0.099
20 N/A 480 0.096
40 0.167 500 0.093
60 0.186 520 0.090
80 0.179 540 0.088
100 0.174 560 0.085
120 0.168 580 0.083
140 0.163 600 0.080
160 0.158 620 0.078
180 0.153 640 0.076
200 0.149 660 0.074
220 0.144 680 0.072
240 0.139 700 0.071
260 0.135 720 0.069
280 0.131 740 0.067
300 0.127 760 0.065
320 0.124 780 0.064
340 0.120 800 0.062
360 0.116 820 0.061
380 0.113 840 0.059
400 0.109 860 0.058
420 0.105 880 0.057
440 0.102 900 0.056
and the absorbance was higher at the 60 second interval than at the 40 second interval.
Table 7
9

0 N/A 460 0.149
20 N/A 480 0.147
40 0.198 500 0.144
60 0.195 520 0.142
80 0.191 540 0.139
100 0.189 560 0.136
120 0.186 580 0.134
140 0.183 600 0.133
160 0.181 620 0.131
180 0.179 640 0.130
200 0.176 660 0.128
220 0.174 680 0.126
240 0.171 700 0.125
260 0.169 720 0.123
280 0.167 740 0.121
300 0.164 760 0.120
320 0.163 780 0.118
340 0.160 800 0.117
360 0.158 820 0.115
380 0.156 840 0.114
400 0.154 860 0.113
420 0.152 880 0.112
440 0.150 900 0.111
and the absorbance never went below 0.100.
Table 8
10

0 N/A 460 0.141
20 N/A 480 0.139
40 0.199 500 0.137
60 0.194 520 0.134
80 0.191 540 0.132
100 0.188 560 0.131
120 0.185 580 0.128
140 0.182 600 0.126
160 0.180 620 0.124
180 0.178 640 0.122
200 0.174 660 0.120
220 0.171 680 0.118
240 0.168 700 0.116
260 0.166 720 0.113
280 0.164 740 0.112
300 0.161 760 0.110
320 0.159 780 0.109
340 0.156 800 0.109
360 0.154 820 0.108
380 0.151 840 0.107
400 0.148 860 0.106
420 0.145 880 0.105
440 0.143 900 0.103
and the absorbance never went below 0.100.
Table 9
11

0 N/A 460 0.084
20 N/A 480 0.082
40 0.180 500 0.079
60 0.174 520 0.076
80 0.168 540 0.074
100 0.162 560 0.071
120 0.156 580 0.069
140 0.150 600 0.066
160 0.146 620 0.064
180 0.140 640 0.061
200 0.135 660 0.059
220 0.130 680 0.057
240 0.126 700 0.055
260 0.122 720 0.053
280 0.117 740 0.051
300 0.113 760 0.049
320 0.109 780 0.047
340 0.105 800 0.045
360 0.102 820 0.043
380 0.098 840 0.041
400 0.094 860 0.039
420 0.091 880 0.038
440 0.087 900 0.036
and it hit the lowest absorbance out of every trial.
Table 10
12

0 N/A 460 0.097
20 N/A 480 0.095
40 N/A 500 0.091
60 0.186 520 0.088
80 0.179 540 0.086
100 0.173 560 0.083
120 0.167 580 0.081
140 0.162 600 0.080
160 0.157 620 0.077
180 0.152 640 0.075
200 0.147 660 0.073
220 0.142 680 0.071
240 0.138 700 0.070
260 0.134 720 0.068
280 0.130 740 0.066
300 0.126 760 0.065
320 0.122 780 0.064
340 0.119 800 0.062
360 0.115 820 0.061
380 0.112 840 0.059
400 0.109 860 0.058
420 0.104 880 0.057
440 0.100 900 0.055
Table 10 shows the absorbance of the crystal violet and sodium hydroxide
solution for 15 minutes. In this trial, the molarity of the sodium hydroxide solution was
0.02 M. There is no data for the 40 second interval because the cuvette was not put into
the colorimeter fast enough.
Table 11
13

0 N/A 460 0.130
20 N/A 480 0.128
40 0.183 500 0.126
60 0.179 520 0.124
80 0.175 540 0.123
100 0.171 560 0.121
120 0.168 580 0.120
140 0.165 600 0.118
160 0.163 620 0.116
180 0.161 640 0.115
200 0.158 660 0.114
220 0.156 680 0.112
240 0.154 700 0.111
260 0.152 720 0.109
280 0.150 740 0.108
300 0.148 760 0.107
320 0.145 780 0.105
340 0.143 800 0.104
360 0.142 820 0.103
380 0.139 840 0.102
400 0.138 860 0.100
420 0.136 880 0.099
440 0.134 900 0.098
0.01 M and the absorbance went below 0.100.
Table 12
14

0 N/A 460 0.126
20 N/A 480 0.123
40 0.185 500 0.121
60 0.180 520 0.118
80 0.177 540 0.115
100 0.174 560 0.112
120 0.170 580 0.110
140 0.167 600 0.107
160 0.165 620 0.106
180 0.163 640 0.106
200 0.159 660 0.104
220 0.157 680 0.104
240 0.154 700 0.103
260 0.151 720 0.102
280 0.149 740 0.101
300 0.146 760 0.100
320 0.144 780 0.099
340 0.141 800 0.097
360 0.138 820 0.096
380 0.136 840 0.095
400 0.133 860 0.093
420 0.131 880 0.091
440 0.128 900 0.090
0.01 M because it was important to see how the rate of the reaction changed when the
concentration of the reactant was changed. In this trial, the absorbance went below
0.100.
Observations:
15
Table 13
Observations for Trials With Sodium Hydroxide Solutions of 0.01 M
Trial Observations (0.01M NaOH)

3 The trial ran smoothly.
4 The absorbance went up a little at the 580 second mark and then back down.
Table 14
Observations for Trials With Sodium Hydroxide Solutions of 0.02 M
Trial Observations (0.02M NaOH)

6 The absorbance was higher at the 60 second mark than the 40 second mark.
10 The cuvette was not placed into the colorimeter fast enough.
Tables 13 and 14 display the observations made when during the trials with their
respective sodium hydroxide solutions. The observations made were if the absorbance
didn’t follow normal trends over time such as increasing at an interval when it was
supposed to decrease. Also, trials with missing data points were also noted due to time
issues. For Trial 4, the absorbance went up during the 580 second interval then started
decreasing again. In Trial 6, the absorbance was higher at the 60 second mark and then
started decreasing, and in Trial 10, there was no data point for the 40 second interval
because the cuvette was not placed fast enough into the colorimeter.
Diagrams:
16
Figure 5. Crystal Violet and Sodium Hydroxide Solution at the Start and End of Trial 1
Figure 5 shows the before and after pictures respectively of the Crystal Violet
and sodium hydroxide solution. As the Crystal Violet and the sodium hydroxide ions
react, the solution undergoes a color fading reaction and goes on to becoming clearer
than purple.
Data Analysis and Interpretation

17
The purpose of this experiment was to determine the rate law of the color-fading
reaction between crystal violet and hydroxide ions. A colorimeter was used to collect the
absorbance of the crystal violet and sodium hydroxide solution, and the data was
displayed on LabQuest. The independent variable in this experiment was the volume of
1% crystal violet solution (ml). The dependent variable was the rate of the chemical
reaction of crystal violet which was affected by the order of the reaction (how the
reaction rate changes based on concentration). The units are s⁻¹, M⁻¹s⁻¹, and M⁻²s⁻¹ for
first-order, second-order and third-order reactions respectively. The control variable in
the experiment was the volume of distilled water (500 ml) used to prepare each solution
and the volume for diluting the solution which was also 500 ml. In the experiment, the
absorbance for the crystal violet and NaOH was monitored, and since absorbance and
concentration are proportional to each other because of Beer’s Law, the molarity (M
which is found by moles/liter) was calculated for both. The data collected was reliable
because it was very consistent, and the same stock solution of crystal violet was used in
each solution. Also, randomization was not possible because the experiment had to be
orderly and systematic. The trials were also replicated the same way each time for
consistency. Descriptive analysis was used to interpret the data because the order of a
reaction can be determined through graphical analysis.
18
Figure 6. Trial 1 Concentration of Crystal Violet Over Time in 0.02 M Solution
19
Figures 6, 7, 8, 9, and 10 above show the natural log of the concentration of
crystal violet over time. The graphs have a linear slope which means that the reaction is
first-order with respect to crystal violet. Also, the slopes of the graphs are negative
because the reactants are depleting over time. Trial 2’s graph was omitted because the
data had the exact same values as in Trial 1. The data was consistent as well in the
sense that each slope is between -0.0013 and -0.0019, which will be useful in coming
up with the rate law.
20
21
22
Figures 11, 12, 13, 14, 15, and 16 show the natural log of the concentration of
crystal violet over time in the 0.01 M solution. The graphs are linear like the graphs of
the 0.02 M solution which indicate a first-order reaction. Each slope had a value
between -0.0005 and -0.0010 except for Trial 5 in Figure 10, so the data was consistent
as well.
The Rate Law:
The rate law is used to determine how the rate of a reaction varies when the
concentration of the reactants change. For the color-fading of crystal violet and sodium
hydroxide, the rate law is:
𝑅𝑎𝑡𝑒 = 𝑘 ′ [𝐶𝑉 + ]𝑛
𝑘 ′ = 𝑘[𝑂𝐻 − ]𝑚
Figure 17. The Pseudo-Rate Law of the Color Fading Reaction
The equation above determines how the color fading reaction changes as the
concentrations of both crystal violet, [CV], and sodium hydroxide, [OH] vary. The k’
value is the pseudo-reaction rate constant because it combines the true value, k, with
respect to sodium hydroxide. The letters n and m correspond with the order of the
reaction with respect to crystal violet and sodium hydroxide. This rate law is only valid if
the concentration of sodium hydroxide ions is greater than the concentration of crystal
violet.
Determining Order of a Reaction:
To determine the order of the reaction, graphical analysis was used because
every order has a distinct feature in their graphs. In a zero-order graph, the
23
concentration over time is linear. For first-order reactions, the natural logarithm of the
concentration over time is linear. Finally, in second-order reactions, the reciprocal of the
concentration over time is linear. This is important because once the order is
determined, the slope of the graph determines the k’ value in the rate law.
Order With Respect to Crystal Violet:
In Figures 6 through 10, the natural log of the concentration of the crystal violet
was shown since they represented linear graphs, indicating that the reaction with the
respect to crystal violet was a first-order reaction. This means that the n value in the
rate law was equal to 1, and the k’ value is equal to the negative slope of the graphs,
which is 0.0015 s-1 since it was the average slope.
Order With Respect to Sodium Hydroxide:
In Figures 11 through 16, even though it was not necessary to determine the rate
law, the concentration of sodium hydroxide was changed to see how the pseudo-rate
constant changed. This was necessary for determining the order of the reaction with
respect to sodium hydroxide as well as for the entire reaction. To see how the pseudo
rate law changed, a ratio of 2 pseudo-rate constants (from Trials 3 of the 0.02 M
concentration and 0.01 M concentration of sodium hydroxide) was made:
0.0013 𝑘[0.02]𝑚
=
0.0007 𝑘[0.01]𝑚
1.8571 = 2𝑚
𝑚 = 𝑎𝑝𝑝𝑟𝑜𝑥. 1 (0.8931)
Figure 18. Determining the Order With Respect to Sodium Hydroxide
24
Figure 18 was used to determine how the pseudo-rate constant changed when
the concentration of sodium hydroxide was half of the initial concentration of 0.02 M.
Because the pseudo-rate constant was almost halved when the concentration of the
sodium hydroxide was halved, the order with respect to sodium hydroxide was 1.
Order and Rate Law of the Color-Fading Reaction:
Now that the values k’, m and n were determined through experimentation and
graphical analysis, both the order and rate law of the whole reaction can be determined.
To find the order of the reaction, the values of m and n must be added in which:
𝑂𝑟𝑑𝑒𝑟 = 𝑚 + 𝑛
𝑂𝑟𝑑𝑒𝑟 = 1 + 1
𝑂𝑟𝑑𝑒𝑟 = 2
Figure 19. Determining Order of the Entire Reaction
The equation above was used to calculate the order of the entire color fading
reaction. Because both orders with respect to crystal violet and sodium hydroxide were
1, the entire reaction ended up being a second-order reaction. This means that when
the concentration of both the reactants are doubled, then the initial rate of the reaction is
quadrupled.
The rate law can now be determined using the simplified version from Figure 14
in which:
𝑅𝑎𝑡𝑒 = 𝑘 ′ [𝐶𝑉 + ]𝑛
𝑅𝑎𝑡𝑒 = 0.0015[𝐶𝑉 + ]1
25
Figure 20. The Rate Law of The Color Fading Reaction
Figure 20 provides the final rate law for the color fading reaction of crystal violet
and sodium hydroxide. The k’ was determined by using the negative slope of the graphs
where sodium hydroxide was held at 0.02 M. The letter n was determined through the
order of the reaction with respect to crystal violet. Sodium hydroxide was not necessary
in this equation because the initial concentration was higher than that of the crystal
violet, allowing this rate law to be valid. Under other conditions, then the pseudo-rate
law would not be acceptable, and the full rate law would have to be used.
Interpretation:
In Figures 6 to 16, the graphs modeled the natural log of the concentration of
crystal violet over time. Figures 6 to 10 were based on the data for the 0.02 M solution,
and Figures 11 to 16 were based on the data for the 0.01 M solution, and all of the
graphs had a linear trend. In Figure 17, the pseudo-rate constant was needed along
with the order with respect to crystal violet. The order of the reaction was found to be
first-order because of the linear trend in the graphs, and because they were first-order,
the pseudo-rate constant was equal to the opposite slopes of the graphs with the 0.02
M solution. In Figure 18 when the concentration of the sodium hydroxide was halved,
the pseudo-rate constant was also halved which meant that the order with respect to
hydroxide ions was first-order. With this information, the order of the entire reaction was
determined to be second-order as shown in Figure 19. Now that the pseudo-rate
constant and order with respect to crystal violet was determined, the rate law was found
in Figure 20.
26
Conclusion
The purpose of this experiment was to determine the rate law of a color fading
reaction between crystal violet and hydroxide. A descriptive analysis was used to
interpret the data as rate order can be determined through graphical analysis. The
original hypothesis stating that when the concentrations of the aqueous solutions of
crystal violet and sodium hydroxide double, the color fading reaction will be a second-
order reaction, was accepted. When finding out the order of the reaction with respect to
crystal violet and sodium hydroxide, both were found to be first-order reactions, making
the entire reaction a second order reaction.
The data collected supported the original hypothesis. For the graphs which
contained the data of the solutions with the 0.02 M concentration of sodium hydroxide,
the natural log of the crystal violet concentration was shown over a 900 second
duration. Each of those graphs displayed a linear trend, and that was important since
first-order reactions have linear trends in their graphs when the natural log of the
concentration is displayed over time. In other words, it meant that the reaction with
respect to crystal violet was a first-order reaction. This also happened with the graphs
for the solutions that had a 0.01 M concentration of sodium hydroxide which indicated
the reaction was also first-order with respect to sodium hydroxide. This meant that the
pseudo-constant value k’ could be found by taking the opposite slope of the graphs.
To prove that the reaction was first-order with respect to sodium hydroxide
mathematically, a ratio of pseudo-constant values was made from 2 trials that had a
different concentration of sodium hydroxide. Figure 18 showed how to calculate for m,
27
which was approximately 1, meaning that the reaction with respect to sodium hydroxide
was a first-order reaction. However, finding out the order of the reaction with respect to
sodium hydroxide was not necessary for finding the rate law.
The pseudo-rate law for the color-fading reaction was found in Figure 20 using
the pseudo-rate constant of the graphs of the solutions with a 0.02 M concentration of
sodium hydroxide as well as the order with respect to the crystal violet. The pseudo-rate
constant was found to be 0.0015 since it was the average opposite slope of all the
graphs. The actual rate law is longer than the one calculated, but the one used in Figure
20 was acceptable because the concentration of sodium hydroxide was greater than the
crystal violet concentration. If this was not the case, the pseudo-rate law would not
work.
The experimental design proved to be vital to the success of the experiment.
Because the procedures were precise and detailed in the Flinn Scientific AP Lab, it was
easy to use them as a base and build up from there. For example, the lab gave step-by-
step procedures to making the stock solution, made up of 1 ml of 25 μM crystal violet
dye and one thousand ml of distilled water as well as how to convert the absorbance of
crystal violet into concentration. However, it did not stop the errors that appeared in a
few of the tests.
Some of the data points were a little inconsistent because at some point during
the experiment, the absorbance would increase then start to decrease. This should not
happen as the absorbance should always be decreasing because reactants always
decrease in reactions to make the products. This is due to the fact that some of the
28
cuvettes had scratches and little marks which could have tampered with the colorimeter.
On some occasions, the cuvettes were not wiped thoroughly because of time restraints
which can be seen with Trial 6 of the solution with the 0.02 M sodium hydroxide solution
which could not be placed in time to get a 40-second reading. Other things could have
affected the colorimeter such as temperature since it affects chemical reactions, and it
could not be controlled.
This research was meant to find out the process of chemical reactions and the
importance of chemical kinetics in the real world. There are so many ideas in chemical
kinetics that help scientists understand how reactions work as well as what can affect
them in significant ways. Since this experiment is so broad, it can be used on other
types of chemical reactions to see how reaction rates work for solutions other than
crystal violet. Scientists can use this research to determine more important things about
chemical kinetics to topics like what factors help rapidly spoil food or how fast certain
medicines can activate. In other words, this research is very flexible with other topics in
chemical kinetics which can help clear up certain process within the world.
29
Works Cited
“Chemical Kinetics.” California State University, Sacramento. Web. 11 Mar. 2018.
“Chemical Kinetics.” Chapter 15: Chemical Kinetics | Chemistry, 3e: W. W. Norton
StudySpace,
Norton Company Inc.,
www.wwnorton.com/college/chemistry/chemistry3/ch/15/chemtours.aspx.
Du, Zhongyu, et al. “Kinetics of the Reaction of Crystal Violet with Hydroxide Ion in the
Critical Solution of 2-Butoxyethanol Water.” The Journal of Physical Chemistry A,
vol. 117, no. 2, 2013, pp. 283–290., doi:10.1021/jp3111502.
“Gram Stain.” Welcome to Microbugz - Gram Stain,
www.austincc.edu/microbugz/gram_stain.php.
Knutson, Theodore R., et al. “A Fresh Look at the Crystal Violet Lab with Handheld
Camera Colorimetry.” Journal of Chemical Education, vol. 92, no. 10, 2015, pp.
1692–1695., doi:10.1021/ed500876y.
Kulesa, Craig. “What Is Spectroscopy?” What Is Spectroscopy?, University of Arizona,
loke.as.arizona.edu/~ckulesa/camp/spectroscopy_intro.html.
Libretexts. “Spectrophotometry.” Chemistry LibreTexts, Libretexts, 21 July 2016,
chem.libretexts.org/Core/Physical_and_Theoretical_Chemistry/Kinetics/Reaction
_Rates/Experimental_Determination_of_Kinetcs/Spectrophotometry.
Mottola, Horacio A., et al. “Absorptiometric determination of hydrogen peroxide in
submicrogram amounts with leuco crystal violet and peroxidase as catalyst.”
Analytical Chemistry, vol. 42, no. 3, 1970, pp. 410–411., doi:10.1021/ac60285a017.
30
“Using the Spectrophotometer.” Michigan State University. Web. 11 Mar. 2018.
“Spectroscopy for Industry.” Diamond Light Source,
www.diamond.ac.uk/industry/Industry-News/Latest-News/Synchrotron-Industry-
News-Focus-Spectroscopy.html.
31

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The Kinetics of Crystal Violet Fading

Patrick Ducusin, Nafshiyat Nihal and Ethan Price

Macomb Mathematics Science Technology Center

Mrs. Hilliard, Mr. Supal and Mrs. Dewey

Data and Observations………………………………………………………………………..11

Data Analysis and Interpretation……………………………………………………………..26

method which helps to classify bacteria.

reactants. Furthermore, this research served as a pathway into the complexity of

Absorbance was measured because tools like spectrophotometers and

colorimeters can only measure absorbance accurately and not concentration of a

can increase the yield of products in certain types of reaction.

more experiments of alternating solutions. However, an alternative solution was made

though it was not needed for the rate law.

different properties of substances.

pH . Crystal violet is a triphenylmethane dye; also known as an intensely colored

Method which is a method of classifying bacteria (Gram Stain).

chemical reactions can be determined by observing the concentration changes over

time. This can be accomplished by using either spectroscopy or colorimetry (Knutson).

Figure 1. The Electromagnetic Spectrum

Spectroscopy is the study of the interaction between matter and electromagnetic

Figure 2. The Mechanism of a Spectrophotometer

absorbed by a certain sample. A spectrophotometer can be used to distinguish

compounds since different compounds absorb different wavelengths, hence, their

to determine the rate law for the dye.

The reaction rate of a chemical reaction is the speed at which a chemical

the molarity (M which is found by dividing number of moles by liters) of a solution

In graphs of concentration v. time, curves are steeper at the beginning of a

units of the rate constant.

The collision of molecules in a chemical reaction are affected by temperature,

which is represented by Eₐ. As temperature increases, the kinetic energy of a molecule

collide in certain orientations, an activation complex (combination of reactants) is

When determining reaction rates, the Arrhenius equation relates activation

together, the Arrhenius equations states that:

Figure 3. The Arrhenius Equation

Figure 3 is the entire equation in which A is equal to the frequency factor, e is

value, the more likely collisions will result in a chemical reaction.

One of which measures the absorbance of hydrogen peroxide in quantities of crystal

mixture is measured against a mixture of the same composition minus hydrogen

measures the absorbance of a mixture composed of a substance and crystal violet.

(Du). This experiment relates to ours because it uses spectrophotometry to analyze a

crystal violet mixture.

hydroxide double, the color fading reaction will be a second-order reaction.

25 µM Crystal violet, (2.5 x 10-5), 50 mL Pipet bulb

Procedures for Calibrating the Colorimeter:

Procedures for Taking Readings:

1. Measure 10.0 mL of 25 μM crystal violet in a serological pipet and add it to a

4. Mix and immediately press “begin timing”.

7. Take the solution out of the colorimeter

8. Measure 10.0 mL of 25 μM crystal violet in a serological pipet and add it to a

12. Mix and immediately press “begin timing”

15. Take the solution out of the colorimeter.

Arrows and CAL

Figure 4. Mechanism of the Colorimeter

calibrating the machine.

Data and Observations

well as the observations for each trial.

Time(s.) Absorbance(Au) Time(s.) Absorbance

Time(s.) Absorbance(Au) Time(s.) Absorbance

Time(s.) Absorbance(Au) Time(s.) Absorbance

Time(s.) Absorbance(Au) Time(s.) Absorbance

absorbance never went below 0.100

Time(s.) Absorbance(Au) Time(s.) Absorbance

Time(s.) Absorbance(Au) Time(s.) Absorbance