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Chemistry

315

Experiment 8:

Kinetics of the Reaction Between
Crystal Violet and Hydroxide
Required Readings

Any brief introduction to chemical kinetics. See the Reserve List in the CHEM 315 syllabus
for full reference citations. Possible options include:

1. Chapter 1, Chemical Kinetics, Atwood.


2. Sections 6.1-6.2, Introduction and The Chemical Reaction Rate, pp. 227-237,
Kellner et al. (First edition).
3. Section 17.2, The Chemical Reaction Rate, pp. 427-437, Kellner et al. (Second
edition).
4. Short Guide to Kinetics (in the Supplementary Reading on Compass).

Overview

Crystal violet (CV+) is a resonance stabilized organic cation. It has a large -conjugated
structure and is often used as a dye, for example in Grams method for classifying bacteria.
It is susceptible to nucleophilic attack at the central carbon atom to afford a leuko (colorless)
form of the dye. In the case of OH, this colorless species is CVOH, a tertiary alcohol.

N N

+ OH- OH

N N N N

CV+ CVOH

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You will investigate the kinetics of this reaction and determine the order with respect to
CV+ and OH, given as x and y respectively in the following equation:
= [ * ], / 0
(1)

Owing to intense color (due to * electronic transitions), CV+ can be conveniently


quantified using UV-vis spectrophotometry and Beers Law, which is given by:
= (2)
A = absorbance (arbitrary units or a.u.)
= molar absorptivity (M-1cm-1)
C = concentration (M)
l= pathlength (cm)
The absorbance is related to the transmittance (the fraction of light that passes through the
sample) by the logarithmic relationship:
9
= 78 = 78 (3)
9:

I0 = source intensity (a.u.)


I = detected intensity (a.u.)
T = transmittance (a.u. or %)

You need to be familiar with both of these equations. Note that transmittance is typically
written as the percentage of light that passes through a sample. So a transmittance nearing
0% (T 0) would be expected for a dark/opaque solution (high A) and a transmittance
approaching 100% (T 1) would correspond to a transparent solution (low A). Thus we
can easily monitor the decrease in [CV+] over time by simply measuring A over that time
period. To simplify the kinetic analysis you will use a large excess of OH ([OH]initial >>
[CV+]initial) rather than one equivalent (can you think why this would matter?).

In addition, you will probe the speciation of CV+ in aqueous solution. Indeed, while neutral
solutions of CV+ are a violet color, those in basic solution are colorless, owing to the
formation of CVOH. Moreover, in acidic media CV+ converts to a green and yellow species,
the predominant form depending on the pH.

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You will have to work efficiently to get your experimental work done in the lab session.
There are three parts to this experiment:
Part A: the molar extinction coefficient of CV+ at 590 nm (590 nm) will be
determined, which will be used to quantify [CV+] in the kinetic runs. The effect of
pH on the electronic spectrum of CV+ will also be investigated.
Part B: the order of the reaction with respect to CV+ will be determined by
collecting kinetic data with [CV+]initial being the experimental variable.
Part C: the order of the reaction with respect to OH will be determined by
collecting kinetic data with [OH]initial being the experimental variable.

Prelab Assignment

1. Look up an MSDS for crystal violet, sodium hydroxide, and nitric acid. What are the
hazards associated with these chemicals? What action should be taken should these
chemicals come into contact with your body (e.g. your skin/eyes)? (1 points)
2. Draw the four major (most pertinent to this experiment) resonance structures of CV+.
(2 points)
3. Why should the same cuvette be used for the blank and sample in this experiment? (1
point)
4. Sketch an absorption spectrum of a yellow solution, labeling your axes and indicating
the wavelength scale in nm. (1 point)
5. CV+ is intensely colored while CVOH, in contrast, is colorless. Why might this be? (2
points)
6. Beers Law does not hold at high concentrations. Give two reasons why this might be.
(2 points)
7. Using a 1 mm cuvette, you measure an absorbance of 0.25 (at 600 nm) for a 2 mM
solution of your compound. What is its molar absorptivity at this wavelength? Use the
conventional units of M-1cm-1 for your answer. (2 points)
8. A sample has an absorbance value of A = 0.260 at 320 nm. What fraction of 320 nm
light was absorbed by the sample? Show your work. (2 points)
9. You have just made a new compound and wish to determine the molar absorptivity at
a given wavelength. How do you go about finding it experimentally? (2 points)

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Materials

Equipment
1 Varian Cary 50 Bio UV-vis spectrophotometer
1 Fisher Science Education pH meter
1 plastic cuvette (pathlength = 1.0 cm)
1 1000-L micropipette (adjustable volume)
1 5 mL volumetric pipette
3 10 mL volumetric flasks
Disposable glass transfer pipettes
Plastic micropipette tips

Chemicals
CVCl (0.100 mM aqueous solution)
NaOH (0.100 M aqueous solution)
HNO3 (16 M aqueous solution)
pH 4 and pH 7 calibration buffer solutions

The crystal violet chloride solution, sodium hydroxide solution and pH buffer solutions can
be located in the cabinet under the bench for experiment 8. The nitric acid is stored in the
cabinet labeled Corrosives beneath the FTIR fume hood, on the wall to the right of
exp 8. Use the small container of HNO3 labeled for this experiment, otherwise points will
be deducted from your final grade. Dispose of all liquid waste in the container designated
for this experiment. Use only water to rinse the glassware. Do not pour waste down the
sink! Plastic cuvettes and pipette tips can be disposed in the container labeled Dye
Contaminated Waste. Dispose of glass in the appropriate glass or sharps waste. Points
will be deducted for any solutions that are not put away by the end of the lab period.

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Part A

Note: all experiments are to be run at room temperature. You will not need to use the Peltier
heating accessory.
1. Place a 1-cm plastic cuvette filled with deionized water in the Varian Cary 50 Bio
UV-vis spectrophotometer. For accurate readings, always wipe the cuvette with
kimtech wipes before placing into the spectrophotometer. Open Simple Reads and
click Setup, changing the wavelength to 590 nm. Click OK and Zero.
2. Using the 0.100 mM CVCl stock solution, prepare 10 mL of 0.0100 mM CVCl in a
volumetric flask (For this experiment, you will always use very dilute solutions to
analyze using UV-vis spectrophotometry. This is to ensure the absorbance readings are
relatively low (< 1). If they were greater, they would be more inaccurate, why might
this be?) Do another dilution by taking 5 mL of this solution and diluting it to 10 mL
to give 0.00500 mM. Repeat this step again so you have solutions of CVCl with
concentrations of 0.0100, 0.00500 and 0.00250 mM. For each sample, find the
absorbance at 590 nm (A590 nm) by placing the sample in the instrument and clicking
Read. Record the absorbance for each in your notebook.
3. Close Simple Reads and open Scan. Click Setup, and change the wavelength
range so it is 750 nm to 350 nm. Click baseline and select baseline correction and
OK. Record a background spectrum by placing a cuvette of deionized water in the
instrument and clicking baseline. To run your sample, simply click Start.
4. Using the 0.100 mM CVCl stock solution, again prepare another 10 mL of 0.0100 mM
CVCl. Record the absorption spectrum of this solution, noting the color of the solution
and the position of your absorption band(s) (max). Now you will study the effect of
acid on CV+. Always calibrate a pH meter before use to ensure accurate readings.
Follow the calibration instructions above the pH meter, or consult a TA for calibration
assistance. You must rinse the pH meter electrode with DI water before and after
each measurement and then (carefully!) pat dry with a kimtech wipe. Do not put
too much pressure on the electrode bulb. Using the Fisher Science Education pH meter
and a scintillation vial, measure the pH of this solution. Adjust pH of the rest of the
solution to 1.5 with concentrated HNO3 and again acquire absorbance data. Be cautious
with the concentrated HNO3, it is located in the cabinet labeled Corrosives under the

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fume hood by the FTIR experiment. Repeat this for pH 0.5. Note: in order to have
enough sample you might have to recycle the solution in the cuvette each time before
adjusting the pH.

Part B

1. You will study kinetics by monitoring 3 reactions in which you vary [CV+]initial while
keeping [OH]initial constant. The solutions must be made up immediately before data
acquisition, which will involve measuring A590 nm every 30 s for a period of 600 s. Use
the 0.100 mM CVCl and 0.100 M NaOH stock solutions and deionized water to prepare
the following mixtures, all of which are 3 mL in volume:

Run VCVCl stock (mL) [CV+]initial (M) VNaOH stock (mL) [OH]initial (M) Vwater (mL)
1 0.500 0.500
2 0.375 0.500
3 0.250 0.500

Fill in the table with the values for initial concentration of the two reactants and the amount
of water needed to make up the solutions to 3 mL. Note: prepare these solutions in a beaker
by first adding water, followed by CVCl solution. Once you have set up the Kinetics
program (see below), add the NaOH solution, quickly mix using the micropipette (draw
the solution up and down the tip a couple of times), transfer to a cuvette, and begin
collecting your data.

2. Close Scan, open Kinetics and click Setup to enter the following parameters:
wavelength = 590 nm, cycle = 0.50 min, stop = 10.00 min. Click Auto Store tab and
select Storage On (Prompt at End), followed by OK and Zero the instrument.
Click Start when you are ready to acquire data. Once the run has been completed,
give your data a filename. Then select File, Save Data As and give it a new name,
saving it as a .CSV file, which can be opened using Microsoft Excel.

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Part C

1. Now you will study kinetics by monitoring 3 reactions in which you keep [CV+]initial
constant while varying [OH]initial. The solutions must be made up immediately before
data acquisition, which will involve measuring A590 nm every 30 s for a period of 600
s. Use the 0.100 mM CVCl and 0.100 M NaOH stock solutions and deionized water to
prepare the following mixtures, all of which are 3 mL in volume:

Run VCVCl stock (mL) [CV+]initial (M) VNaOH stock (mL) [OH]initial (M) Vwater (mL)
1 0.500 0.500
2 0.500 0.375
3 0.500 0.250

Fill in the table with the values for initial concentration of the two reactants and the amount
of water needed to make up the solutions to 3 mL. Note: prepare these solutions in the
same manner as Part B except with the appropriate volumes

2. Repeat the kinetic measurements as in Part B.


3. Clean up your workspace, placing waste into the appropriate containers. Use only water
to clean your glassware.

For your Laboratory Report:

Process your data using Excel. Put your pH observations into a table and any
spectra/graphs in the Data and Results section. Raw data can go in the Supplementary
Information.
Include table in Data and Results or in Supplementary with information on volumes
and concentrations of each component used in runs B1-3 and C1-3
Integrate answers to the Questions below into the Data and Results and Discussion
sections of your lab report.

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Questions:

1. Plot A590 nm versus concentration for CVCl using the three data points obtained as well
as the fact that A590 nm = 0 when C = 0 M. Determine 590 nm by linear regression, paying
close attention to the units of 590 nm and of the quantities plotted on the x- and y-axes.
2. What were your max values at pH 1.5 and 0.5? Qualitatively rationalize the colors that
the solutions appeared (looking at a color wheel might help).
3. In order to process the kinetic data you need to:
a. Convert all A590 nm values to [CV+] by using your value for 590 nm and Beers Law.
b. For each run in Part B:
Plot ln[CV+] versus time (in seconds). Determine the observed rate constant kobs
and its units from the gradient for each run. Using those three kobs values, plot
ln{kobs[CV+]initial} (i.e. the initial rate) versus ln[CV+]initial using the appropriate
[CV+]initial for each run; the gradient represents the order in [CV+] (round the
gradient to the nearest whole number for the order).
c. For each run in Part C:
Plot ln[CV+] versus time (in seconds). Determine the observed rate constant kobs
and its units from the gradient for each run. Using those three kobs values, plot
ln{kobs[CV+]initial} versus ln[OH]initial using the appropriate initial concentrations
for each run; the gradient represents the order in [OH] (round the gradient to the
nearest whole number for the order).
d. Suggest a mechanism for this reaction, drawing chemical structures/curly arrow(s).
Explain how this mechanism reflects the kinetic order for this reaction.
4. Now that you know the order in [CV+] and [OH-], for each run compute the overall
rate constant k. Do this by using the relationship: kobs[CV+] = k[CV+]x[OH]y (you
know [CV+] and [OH] and have x and y from Q3. Give your average value for k, with
the appropriate units. Hence, write the rate law for this reaction.
5. Use the determined rate law to find the rate at which 0.70 mM crystal violet will react
with 0.050 M NaOH.
6. Additional Chemical Engineering question: Describe a new or previously used way to
keep crystal violet from photochemically fading in an industrial setting (perhaps with
large amounts of dye). Besides a dye, what can crystal violet be used for?