CHM151Y

Name: Ruo yi (Caroline) LinStudent #: 1001333112

Section: 112

Experiment #9 performed on February 16th, 2015: Investigating light-matter

interactions.
Abstract
The effect of addition of colour filters in the path of a light beam on the voltage
generated by that beams action on a photocell was examined by placing
successive numbers of filters within the path. Blue, green and a combination of blue
and green filters were used. The effect of the concentration of Copper (II) Sulphate
placed in the path of the light beam on voltage was also examined, using solutions
of varying concentration. The voltage produced by the photocell had an exponential
relationship to the number of filters applied, as well as to the concentration of the
copper (II) sulphate. The absorbance of the filters and solution were found to have
linear relationships to their number and concentration. The voltage-concentration
curves were used to find the concentration of an unknown solution of copper
sulphate. Finally, limitations of the Lambert-Beer law were examined by observing
the photofluorescence of chlorophyll solution. These results confirm the LambertBeer law, while also showing some of its limitations, proving that UV-VIS
spectrometers can be used to help identify various properties of chemical
compounds.
Introduction
Chemical compounds have characteristic absorbance patterns which may be
attributed to their characteristic molecular orbitals. Spectroscopy takes advantage
of these characteristic patterns to identify compounds. One type of spectroscopy is
UV-VIS absorption spectroscopy. When a given frequency of visible light is passed
through a sample, the intensity of the light will be reduced according to the
concentration and composition of the sample. The intensity of an exiting beam of
light may be calculated using the Lambert-Beer law, I=I o10-nloga. In this equation, I is
the intensity of the exiting beam, Io the intensity of the original beam, n the number
of units of material in the path length of the beam, and a some constant greater
than one. The term nloga is the total absorbance of the sample, also denoted by A.
Another formulation of the Lambert-Beer law relates intensity to the path length, b,
of a light beam passing through a solution of concentration c. This equation is
I=bc, where is a constant known as the extinction coefficient. Using the LambertBeer law in conjunction with UV-VIS spectroscopy, one may identify the
concentration of a given solution.
Experimental

CHM151Y
Name: Ruo yi (Caroline) LinStudent #: 1001333112

Section: 112

Figure 1. Experimental setup of a manual spectrometer. The setup used in this

experiment did not employ a diffraction grating or lens. In part A, the holder was
replaced by a series of blue or green filters. In part B, the holder was a cuvette filled
with solutions of Copper (II) Sulphate with varying concentration. The photodiode is
referred to as a photocell in the remainder of the report.
Procedure
A power supply (two 9V batteries) and multimeter were connected to the detector
of the manual spectrometer. The photocell was calibrated to be set in the most
intense region of the spectrum. The background voltage, V s, was recorded. Part A. A
blue filter was placed in the front of the lamp (Figure 1, holder) and the voltage
recorded. Successive blue filters were added and the voltage recorded until all
filters were used. The measurements were repeated using successive additions of
green filters. The measurements were repeated once more using successive
additions of one blue and one green filter each per measurement. Part B. A cuvette
filled with distilled water was placed in the holder position and the voltage of the
photocell recorded. The cuvette was rinsed and cleaned with dry tissue. The cuvette
was filled with a 0.15M solution of Copper(II) Sulphate and placed in the holder
position. The voltage was recorded. The procedure was repeated 9 times with
solutions of varying concentration (see Table 1 under Results). Part C. A spinach leaf
was ground with a few drops ethanol using a mortar and pestle. 10mL ethanol was
added to create a leaf solution. The resultant solution was collected and gravity
filtered using a conical funnel. 10mL ethanol was added to the remaining leaf pulp,
the pulp ground and the rinse gravity filtered. The collected solution was poured
into a vial, which was placed in the holder position of the spectrometer. The vial was
observed at positions both parallel and perpendicular to the path length of the light.
Results and Discussion
Part A: Light filters applied to spectrometer
Filter
#
(Blue)
1

Voltag
e (V)

1.94

Filter
#
(Gree
n)
1

Voltag
e (V)

3.69

Filter#
(Blue+Gr
een)
1

Voltag
e (V)

1.61

Part B: Solutions of
CuSO4
Stock CuSO4 Voltag
#
(M)
e (V)

3.78

CHM151Y
Name: Ruo yi (Caroline) LinStudent #: 1001333112
2
3
4
5
6
7
8
9
10

1
0.65
0.5
0.43
0.4
0.38
0.37
0.36
0.37

2
3
4
5
6
7
8
9
10

3.05
2.54
2.14
1.84
1.58
1.31
1.16
1.02
0.9

2
3
4
5
6
7
8

0.78
0.54
0.44
0.4
0.38
0.36
0.36

Section: 112
1
2
3
4
5
6
7
8
9
10

0.15
0.1
0.07
0.05
0.02
0.01
0.007
0.005
0.002
0.001

1.63
2.23
2.37
2.78
3.17
3.45
3.48
3.52
3.64
3.66

Table 1. The results of the experiment for parts A and B.

Description of qualitative results. When the chlorophyll solution from part C was
observed from a slide parallel to the light beam, it appeared to transmit green light.
However, when the same solution was observed from a side perpendicular to the
light beam, it appeared to transmit red light. The red light may be explained by
photofluorescence. Typically, the energy absorbed by chlorophyll in a plant cell is
used for photosynthesis. However, because the chloroplasts are destroyed when
making the chlorophyll solution, the energy absorbed by the chlorophyll is instead
re-emitted as red light.
When a blue filter is placed in front of the light source, only blue light is transmitted
through the filter. When the same chlorophyll solution is placed in front of the
filtered light, the solution no longer appears red when viewed perpendicular to the
light beam. This is perhaps because the blue filter does not transmit the light
wavelengths which the chlorophyll absorbs.
Sample Calculations. Graphs for absorbance and voltage as a function of number of
filters or concentration of copper (II) sulphate were created using Excel. The value of
Vo was found to be 3.78V from the experiment. The value of V s (voltage from
background light only) was found to be 0.36V. This was the plateau voltage for the
blue and blue+green filter experiments.
The transmittance of a given experimental exposure is given by T=(V- V o)/( Vs- Vo).
For the blue filter experiment with number of filters = 1, T=(1.94-0.36)/
(3.78/0.36)=0.462. The absorbance A is given by
-logT. The absorbance for 1
blue filter is log0.462=0.335. Additional values were calculated with Excel. Graphs
are attached in an appendix.
Additional Discussion. The trendlines in excel use base e rather than base 10. In this
case, the absorbance values indicated by the trendlines are equal to nlna, not
nloga. Notice also that the y intercepts of the absorbance graphs were fixed to 0, as
the absorbance of the filters when there are n=0 should be zero.
The coefficients for the voltage graphs give V o, while the coefficient in front of x (x is
number of filters) gives the absorbance indirectly (note that the value given is for
nlna and must be converted to nloga). The slopes of the absorbance graphs
indirectly give the value of a for the filter experiments (again, a ln to log conversion
must be done), and the extinction coefficients for the concentration experiments.

CHM151Y
Name: Ruo yi (Caroline) LinStudent #: 1001333112

Section: 112

The y intercepts for the absorbance experiments were fixed to 0, as discussed

above.
The voltage of the unknown sample of Copper (II) Sulphate was 3.60. Subtracting
0.36 to correct for background voltage gives 3.24. The exponential trendline which
relates voltage as a function of concentration is Voltage=3.2934e -6.273x, where x is
concentration in M. Plugging 3.24 in for voltage, we get 3.24=3.2934e -6.273x yields
x=0.00261M. Thus, the concentration of the unknown is 2.61x10 -3M.
Conclusions
The Lambert-Beer law in conjunction with a simple spectrometer can be used to find
the concentration of an unknown sample of a given chemical compound. For this
experiment, and unknown concentration of Copper Sulphate which gave a voltage
of 3.60V was calculated to have a concentration of 2.61x10 -3M. Using coloured
filters, the validity of the Lambert Beer law in an experimental setting was
confirmed. Limitations of the Lambert-Beer law were observed using the
phenomenon of chlorophyll photofluorescence. In conclusion, the Lambert-Beer law
is shown to be a useful tool for the identifications of the concentrations of
compounds, although not without limitations.
Post-lab Questions
1. As the number of filters approaches infinity, the term nloga will approach
negative infinity. This means that the intensity of the exiting beam, and
therefore the voltage, should approach 0 (given by taking the limit of I=I o10nloga
as n approaches infinity). In fact, this is exactly what occurs in the blue
and blue-green filter experiments, evidenced by the plateaus in both graphs.
However, the plateau is at 0.36 rather than at 0. It must thus be concluded
that contaminating light from the background is causing the plateau to be at
a higher value than expected.
2. The number of filters is n, and A=nloga, where a is a constant. This equation
has the same form as the equation of a line, y-mx+b, where b=0. Thus,
absorbance is expected to have a linear relationship with number of filters.
Increasing concentration has a similar function to increasing the number of
filters, so it is predicted that the relationship between concentration and
absorbance also be linear.
3. Absorbance has a linear relationship with the number of filters added, so it is
expected that the blue+green absorbance line has a slope equal to the
addition of the blue absorbance slope and the green absorbance slope. This is
actually the case. The slopes for the blue and green lines are 0.2978 and
0.0775 respectively, which add to 0.3753. The actual slope for the
blue+green line is 0.3602.
4. Although the chlorophyll emits red light in all directions, the fluorescence of
the chlorophyll can only be seen viewed perpendicular to the beam. This is
because the intensity of the green light which the chlorophyll transmits is
much higher than the intensity of the emitted light, and when viewing the
chlorophyll from directions where there is much transmitted light passing
through, the emitted light is drowned out by the transmitted light.

CHM151Y
Name: Ruo yi (Caroline) LinStudent #: 1001333112

Section: 112

References
1. Hollas, Michael J. Modern Spectroscopy. John Wiley & Sons, 1996.
2. Sawyer, Heineman, and Beebe. Chemistry Experiments for Instrumental
Methods, John Wiley & Sons, 1984.
3. Skoog, Holler and Nieman. Principles of Instumental Analysis, 5th ed.,
Saunders College Publishing, 1998.

CHM151Y
Name: Ruo yi (Caroline) LinStudent #: 1001333112

Section: 112

Appendix: Graphs of voltage and absorbance as functions of number of

filters/concentration.

Voltage produced by photocell as function of blue light filters

1.8

f(x) = 1.79 exp( -0.59 x )

R = 0.96

1.6
1.4
1.2
1

Voltage (V)

0.8
0.6
0.4
0.2
0

10

12

Number of Filters

Graph 1: Voltage as a function of number of blue filters. x is the number of

filters.

CHM151Y
Name: Ruo yi (Caroline) LinStudent #: 1001333112

Section: 112

Voltage produced by photocell as function of green light filters

3.5

f(x) = 4.03 exp( -0.2 x )

R = 1

3
2.5
2

Voltage (V)

1.5
1
0.5
0

10

12

Number of Filters

Graph 2: Voltage as a function of number of green filters. x is the number

of filters.

Voltage produced by photocell as function of blue-green light filter pairs

1.4

f(x) = 1.74 exp( -0.71 x )

R = 0.97

1.2
1
0.8

Voltage (V)

0.6
0.4
0.2
0

Number of Filters

CHM151Y
Name: Ruo yi (Caroline) LinStudent #: 1001333112

Section: 112

Graph 3: Voltage as a function of number of blue and green filter pairs. x

is the number of filter pairs.

Absorbance as a function of blue light filters

3.000
f(x) = 0.3x
R = 0.99

2.500
2.000

Absorbance

1.500
1.000
0.500
0.000

10

Number of Filters

Graph 4: Absorbance as a function of the number of blue filters. x is the

number of filters.

12

CHM151Y
Name: Ruo yi (Caroline) LinStudent #: 1001333112

Section: 112

Absorbance as a function of green light filters

0.900
0.800
f(x) = 0.08x
R = 1

0.700
0.600
0.500

Absorbance

0.400
0.300
0.200
0.100
0.000

10

12

Number of Filters

Graph 5: Absorbance as a function of the number of green filters. x is the

number of filters.

Absorbance as a function of blue and green light filters

3.000
f(x) = 0.36x
R = 0.99

2.500
2.000

Absorbance

1.500
1.000
0.500
0.000

Number of Filters

CHM151Y
Name: Ruo yi (Caroline) LinStudent #: 1001333112

Section: 112

Graph 6: Absorbance as a function of the number of blue-green filter pairs.

X is the number of filters.

Voltage (V) as a function of Copper (II) Sulphate Concentration

4
3.5
f(x) = 3.29 exp( -6.27 x )
R = 0.99

3
2.5

Voltage (V)

2
1.5
1
0.5
0

0.02

0.04

0.06

0.08

0.1

0.12

0.14

0.16

Concetration of Copper (II) Sulphate (M)

Graph 7: Voltage as a function of the concentration of copper sulphate. X

is concentration.

CHM151Y
Name: Ruo yi (Caroline) LinStudent #: 1001333112

Section: 112

Absorbance as a function of Copper (II) Sulphate Concentration

0.500

f(x) = 0.03 exp( 20.85 x )

R = 0.83
f(x) = 2.89x
R = 0.99

0.450
0.400
0.350
0.300

Absaorbance

0.250
0.200
0.150
0.100
0.050
0.000

0.02

0.04

0.06

0.08

0.1

0.12

0.14

Concetration of Copper (II) Sulphate (M)

Graph 8: Absorbance as a function of the concentration of copper

sulphate. X is concentration.

0.16