Experiment 3

Flash Kinetic Spectroscopy

I. Introduction
Light induced reactions, photochemistry, are some of the most fascinating and important
chemical events. For example, photon absorption initiates chain reactionsphotosynthesisthat
lead to conversion of sunlight into stored energy and to CO
2
uptake form the atmosphere and
conversion to more complex molecules
1
. Photochemistry is also the driving force for
atmospheric chemistry, leading to production of the ozone layer which shields the surface from
high energy UV photons that would otherwise contribute to DNA mutations. One of the most
important advances in the study of photochemistry was the development of flash photolysis by
Norrish and Porter
2
in the late 1940s. Flash photolysis and related flash kinetic experiments
employs a pulse of light (from a laser, lamp, etc.) to initiate a reaction (or sequence of reactions)
with a well defined time zero. After excitation, changes in the sample can be investigated using
the tools of the analytical chemist (of course tailored to the time scale of the events following the
flash).
Time resolved spectroscopies are at the core of the modern physical chemistry laboratory because
they are capable of following processes on the time scale of fundamental reactive events. The
1999 Nobel Prize in Chemistry was awarded to Ahmed Zewail for his work in
Femtochemistry
3
, which is in essence an extension of flash kinetic spectroscopy on the time
scale of a molecular vibration (~10
-15
sec).
N
N
O
O
N
NH
N
N
O
O
N
NH
Figure1: Resonance Structures of 4-anilino-4'nitroazobenzene

2
This experiment was suggested by a J. Chem. Ed. article written by Hair et al.
4
The molecule that
will be studied is 4-anilino-4-nitroazobenzene (4A4N). The cis configuration of this molecule is
shown in Figure 1. It is a push-pull azobenzene with electron donor (-anilino) and acceptor (-
nitro) groups at different ends of a conjugated aromatic system. The presence of these groups
results a resonance structure with an N-N single bond. Contribution from this resonance structure
results in a weakening of the N=N double bond.
In the ground state, trans-4A4N is most stable due to the steric repulsion between the
substituents. Upon irradiation with visible light (the flash) the molecule undergoes a t* t
transition. Following excitation, the molecule is very rapidly (<<1 sec) converted to the ground
state cis isomer. Subsequently, it slowly relaxes back to the more stable trans isomer. The entire
process follows the scheme depicted in figure 2 and detailed below (* denotes an excited state):

1

E
a
4

2

3

Isomerization Coordinate
Figure 2: Potental energy surfaces of 4A4N

E
n
e
r
g
y



trans
cis
0
o
180
o

3
Excitation: trans-4A4N + hu trans-4A4N* (R1)
Excited state isomerization: trans-4A4N* cis-4A4N* (R2)
Relaxation: cis-4A4N* cis-4A4N (R3)
Ground-State isomerization: cis-4A4N trans-4A4N (R4)
While the first three steps of this process occur very quickly, the ground state isomerization
(essentially a rotation about the N=N bond) occurs on a time scale of several minutes. Solvent
and temperature effects on the rate of this isomerization will be studied by flash photolysis.

II. Theory
The rate of ground state isomerization can be written
[cis]
[cis]
d
k
dt
= (1)
where k is the reaction rate constant. This can be integrated to give a familiar exponential decay
expression
t 0 [cis] = [cis]
kt
e

(2)
where [cis]
0
is defined as the concentration of cis-4A4N at time zero. Beers law can be used to
relate concentration and absorbance
A(cis)
t
= cb[cis]
t
(3)
By substitution this gives
A(cis)
t
= A(cis)
0
e
-kt
(4)
Therefore, by fitting the decay of cis-4A4N absorbance to an exponential, the rate constant for
isomerization can be easily determined.



4
In practice, it is much easier to follow the absorbance of trans-4A4N because cis-4A4N does not
contribute to the absorbance at the wavelength of the trans-4A4N t* t transition, while trans-
4A4N does contribute at the wavelength of the cis transition. In order to obtain the isomerization
rate constant, k
isom
, using this absorbance, it is understood that
[trans]

= [trans]
t
+ [cis]
t
(5)

Figure 3: Bleach and subsequent recovery of the absorbance of trans-4A4N in cyclohexane at
40
o
C. Zero time immediately follows the abrupt change in absorbance due to the flash.

Note that the concentration of trans-4A4N after the reaction is completed, [trans]

, will equal the

concentration before the flash. Substituting Eq. 5 into Eq. 4 and applying Beers law:
A(trans)

-A(trans)
t
= A(trans)

-A(trans)
0
e
-kt
(6)
-200 0 200 400 600 800 1000
0.09
0.10
0.11
0.12
0.13
0.14
0.15
0.16


Flash
A
b
s
o
r
b
a
n
c
e
Time (s)

5
Observation of the exponential recovery of the absorbance of trans-4A4N will also give the rate
constant for cis- to trans- isomerization.
This isomerization reaction is strongly dependent on temperature. The rate constant of this
reaction follows the Arrhenius rate equation:
a E
RT
k Ae

= (7)
where A is the frequency factor and E
a
is the activation energy.


1
G. R. Fleming and R. Vangrondelle, Physics Today 27, 48 (1994).
2
R. G. Norrish and G. Porter, Nature 164, 164 (1949).
3
A. H. Zewail, J. Phys. Chem. 100, 12701 (1996).
4
S.R. Hair, G.A. Taylor, and L.W. Schultz, J. Chem. Ed. 67, 709 (1990).

III. Experimental
In this experiment you will be looking at 4A4N which is 4-anilino-4-nitroazobenzene. Noted
that 4A4N is about 25% by weight of the dye Disperse Orange 1. Some of the salts in the dye
will not dissolve in organic solvents, so it may be necessary to decant the solution. Exposure to
ambient light should be minimized after preparation.
In the hood prepare 20mL of a ~10
-6
M solutions of 4A4N in cyclohexane, acetone and
tetrahydrofuran (THF) from the ~10
-5
M solutions provided by the ISF. Before you start, prepare
three scintillation vials by covering them with foil and labeling each vial with the appropriate
solvent. Keep lids on vials closed and store in your drawer when not in use to minimize exposure
to light.
Measure the absorption spectrum of each solution to determine the absorption maximum of the
ground state trans isomer in each solvent. When performing the flash photolysis experiments, the
spectrophotometer should be set at this wavelength. It may be necessary to dilute the solution to
obtain an absorbance less than 1. Detailed instructions on using the SpectroVis can be found at
the end of this lab.
1. Place a sample in the SpectroVis.
2. Begin the run and collect a few seconds of data to provide a baseline.

6
3. Start the isomerization using a flash. Keep the location of the flash as consistent as
possible each run.
4. Wait until the absorbance returns to the baseline value and end the run. (NOTE: the
baseline level may change after the flash so wait until a plateau is achieved)
5. Save your data.
6. Complete this procedure a total of three times for each solvent.
Repeat the experiment at two different temperatures (40C and 60C) in the solvent of your
choice. Record the temperature of the water bath to two decimal places. Temperature dependent
data will allow determination of the frequency factor and E
a
for the reaction.
1. Set your cuvette in the temperature controlled water bath for 60 seconds.
2. Wipe off the cuvette with a kimwipe and place in the SpectroVis.
3. Immediately begin the run. Start the isomerization using a camera flash and wait until the
absorbance returns to the baseline to end the run. Being efficient once the cuvette is
removed from the water bath is essential to getting good data.
4. Complete this procedure a total of three times at each temperature.

IV. Analysis
You will find it useful to use a program such as Origin or Excel in interpreting your results.
1. Plot the transient absorption traces and determine k for the reaction in each solvent at
room temperature. Report these values in tabular form.
2. The polarity of the solvents increases in the following order: cyclohexane < THF <
acetone. Explain how solvent polarity affects the isomerization rate? (Hint: What affect
will solvent polarity have on the resonance structures of 4A4N? How will this affect the
N=N bond?)
3. Qualitatively sketch the effect of solvent polarity on the ground state potential energy
surface.

7
4. Using the temperature dependent data, determine the activation energy and frequency
factor for isomerization in your chosen solvent. Report these values.
5. How large is E
a
compared to RT? Discuss the time scale of the reaction in terms of this
answer.

8
SpectroVis Instructions: Flash Kinetics

Preparing the instrument:
1. Turn on the Vernier LabQuest. Use the provided power cable to plug it into the wall.
2. Connect the SpectroVis Plus Spectrophotometer to the LabQuest with the cable included
(the USB side of the cable goes into the LabQuest2).
Data Collection:

Complete the following three sections completely for one solvent set before proceeding to
the next solvent set.

Spectrometer calibration
1. Click on (the Meter tab) to get to the correct screen.
2. Click on Sensors with the stylus.
3. Choose Calibrate Spectrometer. NOTE: recalibrate for each solvent.
4. Place the Blank sample in the cuvette holder, making sure that the cuvette is placed so
that the light source passes through the clear side.
5. Follow instructions in the dialog box to complete the calibration. DO NOT skip the
Lamp Warm-Up. (If you are recalibrating later, when the spectrometer has been on for a
while, you may skip the lamp warmup.)
6. Click Finish Calibration.
Absorption Spectrum
1. Place your cuvette containing a sample of ~10
-6
M 4A4N in the cuvette holder
2. Press the button to start recording data
3. Determine the wavelength that corresponds to the absorption maximum
4. Save your data. You will need to do this for each solvent.
Kinetics Run Collection
1. Click on Sensors Data Collection
2. Under the Mode drop down menu select Time Based
3. Set the rate to 1 sample/s and the time to 2000s and press OK
4. Click on the large red area and select Change Wavelength. Enter in the wavelength you
wish to monitor (the wavelength of maximum absorption).
5. Insert the cuvette and press the button to start recording data.
6. Collect a few seconds of data to provide a baseline then use a flash to start the
isomerization.
7. Save each run with your drawer number and sample name. You will not export any data
until the end of the lab


9
Extracting Data to USB:
1. Plug in your USB drive
2. Go to File Open select one of your data files
3. Go to File Export and then click on the USB icon and name the file

Be sure it is a .txt file or you will not be able to read it on anything but the LabQuest, and it may
be impossible for you to retrieve your data after you leave the laboratory for the day. You should
be able to open/import this .txt file into excel in order to graph and analyze the data.