Verification of Beer-Lampbert Law

By
J. Evans, J. Eldridge, D. Lockhart
Instrumental Analysis Laboratory
CHEM 310
Spring 2011
Date of Experimentation:
January 31, 2011
Date of Turn In:
February 10, 2011

ABSTRACT:

A Spectrophotometer sends light from a light emitting diode (LED) through a solution
placed in a cuvette inside the Spectrophotometer. The light that passes through the solution
strikes a photodiode. A higher concentration of the colored solution absorbs more light and
transmits less light than a solution of lower concentration. In this lab we planned to test this very
theory by using Mn and different mixtures of it. Each mixture was read in a spectrophotometer.
From previous studies we were lead to believe that once we found the absorbance we would be
able to use the Beer-Lambert equation to find the concentration of the solution. When we added
light solutions to the mixture, we would see that the absorbance of light increased. We were
surprised to see that adding Cr increased the amount of light almost as much as light. It was
found that using water increases the absorbance by 1.2 % in many cases. After using the
Spectrophotometer we were able to graph the data and find a slope and equation of the line for
each sample. We were then able to use this to find the concentration of each sample.

AIM:

(i) How to determine absorbance spectrum of unknown compounds (qualitative analysis).

(ii) How to determine the λmax of a compound.
(iii) How to use an absorbance spectrum to identify a pure compound.
(iv) How to use the Beer-Lambert Law to determine the concentration of compounds in solution.
(v) How to prepare and graph standard or calibration curves for quantitative analysis using
colorimetry.
(vi)To verify Beer’s law of absorption of light for some solutions
 I. Introduction

A Spectrophotometer sends light from a

light emitting diode (LED) through a solution
placed in a cuvette inside the
Spectrophotometer. The light that passes
through the solution strikes a photodiode. A
higher concentration of the colored solution
absorbs more light and transmits less light than
a solution of lower concentration. The
Spectrophotometer monitors the light received
by the photodiode as either an absorbance or a
percent transmittance value. You can use the
amount of light that penetrates the solution and
strikes the photodiode to compute the
absorbance of each solution. A graph of
absorbance versus concentration for a series of
standard solutions shows a linear relationship.
The direct relationship between absorbance and concentration for a solution is known as Beer’s
law.

I0
A = constant x c = log 10
I

In Beer’s law, A is the absorbance, c is the concentration, I0 is the intensity of radiation before
passage through the solution, and I is the intensity of radiation transmitted through the solution.
Spectrophotometers induce transitions from the ground state of an atom, ion or molecule
to an electronically excited state of the same atom, ion or molecule, by complex processes
involving variations in the energy of more than one electron. The “ONE-ELECTRON
APPROXIMATION” can however serve as a useful qualitative description of the electronic
processes which give rise to absorption of ultra-violet and visible light by atoms, or molecules.
The absorption of a monochromatic beam of light by a homogeneous absorbing system is
described well by the Beer-Lambert Law:

I
=10 −εcl (1)
IO

where I0 represents the light energy (or number of quanta)of strictly monochromatic light
incident per unit of time at the front of a column of a single absorbing species of concentration c,
moles/litre; I is the energy per unit time transmitted through the column of material, 1 cm length,
and , extinction coefficient.

In this experiment manganese and chromate samples were provided. The experiment entails
verifying Beer's Law by measuring the absorbance versus concentration. A straight line plot
should result if the solution obeys Beer's Law.

Beer-Lambert Absorption Law

The relationship between the amount of light a solution transmits (or absorbs), and its
concentration is given by Beer's Law:

I0
log =εcl
I (3)

Where I0 is the intensity of the light incident on the solution

I is the intensity of the light transmitted by thesolution
c is the concentration (moles/litre) of the solution
l is the path length (in cms), i.e. the length of the path of solution through which the
light passes
 is the decadic molar extinction coefficient, a characteristic of the components of
the solution.
is the absorbance of the solution
I0
log
I

A spectrophotometer measures the absorbance of a medium at any wavelength.

II. Experimental Methods and Materials

A. Apparatus:
• STANDARD CURVES
• Spectrophotometer/ colorimeter
• Standard flasks, etc
• Two identical cuvettes with a known thickness.

A. Experimental set up:

 STOCK SOLUTIONS
• 500 ml of a solution (Mns) of KMnO4 solution containing [Mn] = 0.1 g/l1
• 500 ml of a solution (Crs) of K2Cr2O7 (or Na2Cr2O7) solution containing
○ [Cr] = 0.6 g/l1
• Dilute Mns with water to make 100 ml of each of the following:
○ MnA1: [Mn] = 0.050 g/l
○ MnA2: [Mn] = 0.040 g/l
○ MnA3: [Mn] = 0.030 g/l
○ MnA4: [Mn] = 0.020 g/l

Standard Solutions
• Dilute 20 ml of each of the four solutions (MnA1 - MnA4) with enough water to obtain 50.0
ml of each of four solutions: MnB1 - MnB4, respectively.
• Dilute 20 ml of each of the four solutions (MnA1 - MnA4) with 20.0 ml of Crs then
complete the addition of water to obtain 50.0 ml of each of four solutions: MnCrB1 -
MnCrB4, respectively.
• Dilute 20 ml of Crs to 50.0 ml to make CrB.

A. Determine the Absorbance and %T at 525 nm for each of the following samples:
• the four MnB1 - MnB4 solutions using water as a blank2
• the four MnCrB1 - MnCrB4 solutions using water a a blank2
• the four MnCrB1 - MnCrB4 solutions using CrB as a blank2
• CrB using water as a blank2
2. One a single sheet of graph paper plot % Transmittance (as the ordinate) versus:

• [Mn] in the four MnB1 - MnB4 solutions using one color or symbol
• [Mn] in the four MnCrB1 - MnCrB4 solutions using a different color or symbol
• [Cr] as a straight horizontal line

3. One another sheet of graph paper plot Absorbance (as the ordinate) versus:

• [Mn] in the four MnB1 - MnB4 solutions using one color or symbol
• [Mn] in the four MnCrB1 - MnCrB4 solutions (using water as a blank) using a different
color or symbol
• [Mn] in the four MnCrB1 - MnCrB4 solutions (using CrB as a blank) using a different
color or symbol than either of the other Absorbance plots
• [Cr] as a straight horizontal line

Procedure:
1. Let the instrument “warm up” for at least 15 minutes.
2. Set the desired wavelength (525nm) with the wavelength control.

2
3. Set % transmittance (top scale) at zero, or the absorbance (bottom scale)at infinity using the
amplifier control, A.
4. Place a test tube at least half filled with water or the appropriate solvent in the sample holder
(sometimes called the blank). Be sure that the sample holder is in place by pressing down on it.
The top of the holder must be shut whenever you adjust the instrument or take readings, to
prevent stray light from entering the instrument.
5. Set the needle at zero absorbance (or 100% transmittance) using the light control.
6. Remove the blank. The needle should swing to infinite absorbance (or zero % transmittance).
If it does not, repeat steps 3 through 6.
7. The instrument is now correctly calibrated. To take a reading, place a test tube containing your
sample in the machine and close the lid; after 10 seconds record the absorbance (and wavelength
if necessary).
8. Before taking another reading on the same or different sample, you should again check that the
absorbance scale reads infinity with nothing in the sample holder and that it reads zero with the
blank sample.
I. Results:
Table 1. Absorbances and Transmittance of
KMnO4
%
Samples Absorbance transmittance
KMnO4
0.0200 0.32 48.31
0.0160 0.50 31.33
0.0120 0.66 21.98
0.0080 0.81 15.45

KMnO4
.02 H20 0.37 43.15
.016 H20 0.56 27.42
.012 H20 0.76 17.38
.008 H20 0.95 11.30

KMnO4
.02
Chromium 0.33 47.10
.
016Chromi
um 0.52 29.99
.012
Chromium 0.72 18.97
.008
Chromium 0.91 12.33

Cr
A1 0.036 92.045
Figure 1 Absorbance of Mn by Spectrometer

Figure 2 Transmittance of Mn and Cr by Beer’s Law

II. Analysis of Results

Absorbance = -log (percent transmittance/100)
A= -log (%T/100)

%T= 100 * 10^-A

100 * 10 ^ - 0.5620 = 27.4157%

0.1g Mn1L × 1 mol Mn54.9 Mn × 1 mole KMnO41 mol Mn ×158.034 g1 mol

KMnO4=0.288gl÷2 =.144gKMnO4l

MnA1: [Mn] = 0.050 g/l

Mnb1: [Mn] = 0.050 g/l * (20ml/50ml)= .020g/l

III.Conclusion:

The extinction coefficient (ε) in the Beer-Lambert equation is a measure of how strongly
a compound in solution absorbs light at a particular wavelength. Extinction coefficients are
useful numbers that allow researchers to calculate the concentration of a compound in solution
(quantitative analysis). We found that the darker the solutions the least amount of light that was
able to get through. When we added light solutions to the mixture, we would see that the
absorbance of light decreased. We were surprised to see that adding Cr increased the amount of
light almost as much as light. It was found that using water increases the absorbance by 1.2 % in
many cases. After using the Spectrophotometer we were able to graph the data and find a slope
and equation of the line for each sample. We were then able to use this to verify Beer’s Law. In
short, based on the data that was collected we were able to determine that the absorbance was
proportional to the concentration; percent transmittance is inversely proportional to the
concentration using a logarithmic relationship.
References:
1. J. D. J. Ingle and S. R. Crouch, Spectrochemical Analysis, Prentice Hall, New Jersey

(1988

2. ISBN 0521339561 Houghton, J.T. The Physics of Atmospheres 2nd ed. Chapter 2