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ABSORPTION LAWS (QUANTITATIVE

ANALYSIS)
Belay Haile, MSc (Analytical Chemistry), PhD Scholar (Postharvest
Technology), Arba Minch University, CNS, Chemistry Department,
Arba Minch Ethiopia February 2021
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Absorbance of Electromagnetic Radiation

• In absorption spectroscopy a beam of electromagnetic
radiation passes through a sample.

• Much of the radiation is transmitted without a loss in

intensity. At selected frequencies, however, the radiation’s
intensity is attenuated. This process of attenuation is called
absorption.

• Two general requirements must be met if an analyte is to

absorb electromagnetic radiation.
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Cont. …
• The first requirement is that there must be a mechanism by
which the radiation’s electric field or magnetic field interacts
with the analyte.

• For ultraviolet and visible radiation, this interaction involves

the electronic energy of valence electrons. A chemical bond’s
vibrational energy is altered by the absorbance of infrared
radiation.

• The second requirement is that the energy of the

electromagnetic radiation must exactly equal the difference in
energy, ∆E, between two of the analytes quantized energy
states.
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Cont. …

Energy level diagram showing

difference between the
absorption of:

Infrared radiation(left) and

Ultraviolet–visible radiation
(right).
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Transmittance and Absorbance

Transmittance is defined as the ratio
of the electromagnetic radiation’s
power exiting the sample, PT to that
incident on the sample from the
source, P0.

…………..(1)

Schematic diagram showing the

Multiplying the transmittance by
100 gives the percent transmittance
(%T), which varies between
 100% (no absorption) and
 0% (complete absorption).
attenuation of radiation passing through
a sample; P0 is the radiant power from
the source and PT is the radiant power
transmitted by the sample
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Cont. …
• An alternative method for expressing the attenuation of EM
radiation is absorbance, A, which is defined as:
……………………….. (2)

EXAMPLE: A sample has a percent transmittance of 50.0%.

What is its absorbance?

SOLUTION
With a percent transmittance of 50.0%, the transmittance of
the sample is 0.500. Substituting into equation gives
A = –log T = –log(0.500) = 0.301
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Absorbance and Concentration: Beer’s Law

• When monochromatic EMR passes through
an infinitesimally thin layer of sample, of
thickness dx, it experiences a decrease in
power of dP.
• The fractional decrease in power is
proportional to the sample’s thickness and Factors used in deriving the Beer–
the analyte’s concentration, C; thus Lambert law

………….…..(3)

where P is the power incident on the thin

layer of sample, and α is a proportionality
constant.
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Cont. …..
• Integrating the left side of equation (3) from P = P0 to P= PT ,
and the right side from x = 0 to x = b, where b is the sample’s
overall thickness

gives

Converting from ln to log, and substituting equation (2), gives

………………………… (4)

where a is the analyte’s absorptivity with units of cm–1 conc–1

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Cont. ……
When concentration is expressed using molarity, the absorptivity is
replaced by the molar absorptivity, ε (with units of cm–1 M–1 )

…………………………………….. (5)

The absorptivity and molar absorptivity give, in effect, the probability

that the analyte will absorb a photon of given energy. As a result, values
for both a and ε depend on the wavelength of electromagnetic radiation.

Equations (4) and (5), which establish the linear relationship between
absorbance and concentration, are known as the Beer–Lambert law,
or more commonly, as Beer’s law.

Calibration curves based on Beer’s law are used routinely in

quantitative analysis.
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Cont. …
 
•EXAMPLE: A 5.00 10–4 M solution of an analyte is placed in a
sample cell that has a pathlength of 1.00 cm. When measured
at a wavelength of 490 nm, the absorbance of the solution is
found to be 0.338. What is the analyte’s molar absorptivity at
this wavelength?

SOLUTION
Solving equation (5) for ε and making appropriate
substitutions gives
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Beer’s Law and Multicomponent Samples

• Beer’s law can be extended to samples containing several

absorbing components provided that there are no
interactions between the components.
• Individual absorbances, Ai, are additive.
• For a two component mixture of X and Y, the total
absorbance, Atot, is

Generalizing, the absorbance for a mixture of n components,

Am , is given as
………………………. …. (6)
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Limitations to Beer’s Law

oAccording to Beer’s law, a calibration

curve of absorbance versus the
concentration of analyte in a series of
standard solutions should be a
straight line with an intercept of 0
and a slope of ab or εb.

oIn many cases, however, calibration

curves are found to be nonlinear Calibration curves showing positive and
negative deviations from Beer’s law.
oDeviations from linearity are divided
into three categories: fundamental,
chemical, and instrumental.
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Fundamental Limitations to Beers Law

• Beer’s law is a limiting law that is valid only for low concentrations of
analyte
There are two contributions to this fundamental limitation to Beer’s law.

At higher concentrations the individual particles of analyte no longer

behave independently of one another. The resulting interaction
between particles of analyte may change the value of ε.

A second contribution is that the absorptivity, a, and molar

absorptivity, ε, depend on the sample’s refractive index. Since the
refractive index varies with the analyte’s concentration, the values of a
and ε will change.

 For sufficiently low concentrations of analyte, the refractive index remains essentially
constant, and the calibration curve is linear.
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Chemical Limitations to Beer’s Law

• Chemical deviations from Beer’s law can occur when the
absorbing species is involved in an equilibrium reaction.

• If both HA and A– absorb at the selected wavelength, then Beers

law is written as
…………………………….……..… (7)

where CHA and CA are the equilibrium concentrations of HA and A–.

……………. (8)

Where, Ctot = CHA + CA

Depending on the relative values of εHA and εA , the calibration
curve will show a positive or negative deviation from Beer’s law
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Instrumental Limitations to Beer’s Law

There are two principal instrumental limitations to Beer’s law.

• The first limitation is that Beer’s law is strictly valid for purely
monochromatic radiation; that is, for radiation consisting of only
one wavelength.

 Using polychromatic radiation always gives a negative deviation from Beer’s

law,
• Stray radiation is the second contribution to instrumental
deviations from Beer’s law. (stray radiation: Any radiation reaching the
detector that does not follow the optical path from the source to the detector.

 Stray radiation arises from imperfections within the wavelength selector