Applications

& Limitations of
Beer Lambert Law

Presented By:
Irsa Jamal

Presented to:
Dr. Khalid Hussain
According to the law of Lambert,
“When a ray of monochromatic light passes
through an absorbing medium its intensity
decreases exponentially as the length (l) of the
absorbing medium increases”.

I = Io e-kl ……… (1)

Beer's law states that
“The amount of light absorbed is proportional to the number of
chromophores present in the medium that the light is passing through”.
In other words, “the amount of absorbed light is proportional to the
concentration of the absorbing species (chromophores)”.
‘or’
“When a monochromatic light passes through an absorbing medium its
intensity decreases exponentially as the concentration (c) of the
absorbing medium increases”
I = Io e-kc ……… (2)
These two laws are often combined into what is often referred to as Lambert-Beer's, Beer-Lambert's or simply
Beer's law:
“The absorbance of a solution is linearly proportional to its concentration c, and the path length l of the light in
the solution”
I = Io e-kcl ……… (3)
A = log Io / I = kcl ……… (4)
Where,
A = Absorbance or optical density or extinction co-efficient
k = proportionality constant
c = concentration
l = path length
Io = incident light
I = transmitted light
When the path length is measured in centimeters, and the concentration of the absorbing species is measured in
Molarity, the proportionality constant is called the Molar Absorptivity ‘ε’, having units of M -1cm-1,
Absorbance = εcl ……… (5)
ε = molar absorptivity (molar extinction coefficient)
Application of Beer lambert Law:
The absorption of UV and visible light is quantitatively described by the laws of Lambert and Beer. [1]
Limitations of Beer lambert Law:
The proportionality between absorbance and concentration inferred by Eq. (5) is not always observed, causes for
deviation from Lambert-Beer's law may be of chemical as well as instrumental origin

 The light used must be monochromatic, or otherwise for two different wavelengths (λ1, λ 2) two values of
absorbance will be obtained.

Explanation: Experimentally it is not possible obtain monochromatic radiation. In practice the sample is
exposed to polychromatic radiation of a certain wavelength range, the more narrow the band of wavelengths,
the better since polychromatic radiation leads to deviations from Lambert-Beer's law.

 The law is not applicable for highly concentrated solutions. Higher concentrations of some other colourless
salts even adversely affect absorption of light.

Reason: At high drug or analyte concentrations (typically >0.01 M) deviations from linearity may be
observed due to refractive index changes and because the close proximity of the absorbing molecules will
affect their charge distribution and lead to alterations in their absorptivity. Furthermore, the com-pounds
should not take part in reactions, such as self-association reactions and chemical degradation which may lead
to changes in absorbance.
Limitations of Beer lambert Law:
 Particles present in the sample will also lead to deviations from Lambert-Beer's law due to light scattering.

 If a dilute solution during measurement undergoes chemical reaction such as hydrolysis, association,
dissociation or polymerisation then the law is not valid.

 Deviation from Lambert-Beer's law can also occur due to stray light. This is light that reaches the detector
without having passed through the sample due to light scattering within the instrumentation or light entering
from outside the instrument.

 If a system conforms to Beer's law, a graph of absorbance against concentration should be a straight line
passing through the origin. In case Beer's law is strictly adhered to we may get straight line graph or
calibration curve.
 REFERENCES
 “Analytical Techniques in the Pharmaceutical Sciences”Edited by Anette Mullertz, Yvonne Perrie, and
Thomas , Chapter 1, page no. 4
https://books.google.com.pk/books?id=a5fvDAAAQBAJ&pg=PA4&dq=lambert+beer+law&hl=en&sa
=X&redir_esc=y#v=onepage&q=lambert%20beer%20law&f=false

 
 “Practical Materials Characterization” Edited by Mauro Sardela,, Chapter 2 , Page 59
https://books.google.com.pk/books?id=umEgBAAAQBAJ&pg=PA59&dq=uses+of+beer+lambert+law&hl=en&sa=X&redir_esc=y#v=onepage&q=
uses%20of%20beer%20lambert%20law&f=false

 “Basic Concepts Of Analytical Chemistry” Edited by, S M Khopkar, Chapter 20, page no. 210
https://books.google.com.pk/books?id=e8Ju_n8DN1sC&pg=PA210&dq=lambert+beer%27s+law
&hl=en&sa=X&ved=0ahUKEwja-MC4zvDPAhXLQY8KHdFBD20Q6AEIKTAC#v=onepage&q=lambert%20be
er's%20law&f=false