CH2213: Analytical Chemistry

Lecture -19
Spectroanalytical Chemistry:
Absorbance Spectroscopy
Professor Manickam Jayakannan
Department of Chemistry
IISER Pune, INDIA
E-mail: jayakannan@iiserpune.ac.in
Website: http://www.iiserpune.ac.in/~jayakannan/

Date: 28th March 2022

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 O

COOH R
N

q Spectroscopic analytical methods are based on measuring the amount of

radiation produced or absorbed by molecular or atomic species of
interest.
q Spectroscopic methods according to the region of the electromagnetic
spectrum used or produced in the measurement. The g-ray, X-ray,
ultraviolet (UV), visible, infrared (IR), microwave, and radio-frequency
(RF) regions have been used.

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Energy Level Diagrams

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 Analyte interaction with Electromagnetic Spectrum
H !*
C
(OR) C---H 1H (OR) 13C

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Electronic Absorption

LUMO

LUMO

HOMO

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 The Absorption Process

The absorption law, also known as the Beer-Lambert law or just Beer’s law, tells us
quantitatively how the amount of attenuation depends on the concentration of the
absorbing molecules and the path length over which absorption occurs.
A monochromatic radiation as it passes through an absorbing solution of thickness
b cm and concentration c moles per liter; because of interactions between the photons
and absorbing particles, the radiant power of the beam decreases from P0 to P.

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The Absorption Process

The terms P0 and P will henceforth refer to the

power of a beam that has passed through cells
containing the solvent (or blank) and the
analyte solution, respectively.

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 Absorption Spectrophotometer

Instrumentation

S1

S0

Abs
Onset

Wavelength (nm) 9
Determination of Molar Extinction Coefficient

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 Molar Extinction Coefficient: Importance

Dyes Absorbance Extinction Coefficient

For compounds with high Extinction Coefficient, Low concentration of probe

dye is sufficient enough for detection

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 Colorimetric sensing of Metal ions

Ref: Wang et al. RSC Adv., 2019, 9, 6643

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Colorimetric sensing of Metal ions

2 : 1 Ligand-to-metal ion Complex

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Electronic Transitions: Vapor & Solution State

S1

v3
v2
v1
v0

S0

v3
v2
v1
v0

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 Rigid- Aromatic Structures

250 300

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Application of Absorbance Spectroscopy

qDetection of Toxic molecules, Heavy Metal ions, etc

qKinetics of Reaction
qDrug Release Profile Kinetics
qIsomerization
qStructural Changes
qIntermolecular Interactions (Aggregates H and J-type)
qSupramolecular Assemblies: Micelles, Molecular Boxes

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