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Atomic Spectroscopy
• Atomic Emission
• Atomic Absorption
• Atomic Fluorescence
• Use of X-rays
Molecular Spectroscopy
• Fourier Transform Infra-red (FT-IR) Syllabus Recap
• Organic UV-Vis Absorption
• Effect of Solvent
• Inorganic UV-Vis Absorption
• Qualitative Analysis
• Quantitative Analysis
• Molecular Fluorescence
• Equipment for Molecular Spectroscopy
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MOLECULAR
SPECTROSCOPY
CDB 3093: ANALYTICAL CHEMISTRY
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Outline
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Learning Outcomes
At the end of the chapter:
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Introduction
Molecular spectroscopy based on ultraviolet, visible and
infrared radiation is widely used for the identification and
determination of many inorganic, organic and
biochemical species.
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Fourier Transform Infra-Red (FT-IR)
Identity of compounds
Presence or absence of functional groups
(C=O,C=C,C-H, C≡C or O-H)
Structure of molecules.
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Fourier Transform Infra-Red (FT-IR)
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Recall: Molecular Absorption
The energy E associated with the bands of a
molecule is made up of three components.
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Infrared Regions
i) Near IR -4000-14000 cm-1
- for routine quantification determination of certain
species, such as water, CO2, sulfur, HCs, amine
nitrogen.
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Abbreviation Table of Group Frequencies for Organic
Functional Groups
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Example: IR spectrum for n-butanal (n-butyraldehyde)
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Examples: IR spectrum for other compounds
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Identify the molecular structure based
on this IR spectrum: Example 1
? ?
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Identify the major peaks
C-H
N≡C
C≡C
C-O
C-N
C=O O-H
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Answer
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Example 2
C-H
C-H
CH3-(CH2)8-CH3
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Example 3
Transmittance, %
Wavenumber, cm-1
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Example 4
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0
4000 3000 2000 1500 1000 500
Wavenumber, cm -1
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Example 6
The spectrum below is that of a nitrogen-containing
substance and has a molecular weight of about 55. Sketch
the possible molecular structure of this substance?
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Outline
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Ultraviolet and Visible (UV/Vis)
Absorption Spectroscopy
Are widely used for all quantitative analysis in
chemical, environmental, forensic and clinical
laboratories.
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UV/Vis Spectrum
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Three different aspects of UV/Vis spectra
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Molecular Spectroscopy
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Organic UV/Vis Absorption: Kind of
Transitions
All organic compound contain valence electrons that can be
excited to higher energy level. transitions of electrons.
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• (3) Electron in pi orbital (π) double or triple bonds.
These electrons are most readily excited and are
responsible for a majority of electronic spectra in the
UV/Vis regions.
σ bonding
π bonding
n bonding
σ* anti-bonding
π* anti-bonding
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• The most common transitions are from π or n to
antibonding π* orbitals, and these are represented by
π π* and n π* transitions, indicating a transition to
an excited π* stage.
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1) σ σ* transition
- appears in the far UV, λmax <150 nm
- requires a significant energy.
- adsorption corresponding to breaking of saturated
carbon: C-C, C-H, C-O, C-X and etc.
- not observed in a normal UV/Vis work.
- Example: Hexane (gas state) : λmax = 135 nm
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2) n σ* transition
- promotion of n electron from an atom of O, N, S, Cl to
an σ*.
- Adsorption range: 150-250 nm.
- Example: 180 nm for alcohols, 190 nm for ethers or
halogen derivatives and in the region of
220 nm for amines.
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3) n π* transition
- usually observed in molecules of an unsaturated group
(halogen).
- Example: carbonyl band (C=O) 270-295 nm
4) π π* transition
- compounds possessing an isolated ethylenic double bond
(C=C) strong adsorption band ~ 170 nm.
- Example: Ethylene: λmax = 165 nm
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Examples: Typical Absorption Spectra
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Absorption characteristic of some common
Chromophores
Molecules containing unsaturated organic functional
groups and capable of absorbing UV/visible radiation.
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Molecular Spectroscopy
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Effect of Solvent
The position and intensity of the absorption bands will
vary with the nature of the solvent used each
solvent has its own characteristic polarity.
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Common solvent for UV/Vis Region
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Molecular Spectroscopy
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Inorganic UV/Vis Absorption:
Absorption by Organic Compounds
A number of inorganic anions exhibit UV absorption
bands.
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Examples:
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Qualitative Applications of UV/Vis
Absorption Spectroscopy
Detecting chromophoric groups.
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Quantitative Analysis
UV/Vis absorption spectroscopy most useful
tools available for quantitative analysis.
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Method of Analysis
1) External Standard (calibration curve)
2) Standard Addition
i) Single-point addition
ii) Multiple additions
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External Standard (Calibration Curve)
A standard solutions of the analyte is used to
construct a calibration curve of absorbance versus
concentration.
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Example : The method of external standards
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Determine the concentration of unknown:
y = 0.0302 x -0.001
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Standard Addition Method
For those samples having difficulties in the production of
standards with an overall composition.
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Example: Single-point Addition
Method
The single-point standard solution addition method was used
in the determination of phosphate by the molybdenum blue
method.
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Solution
The absorbance of the first solution is given by (Beer’s Law):
A1 = εbcx
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Solution
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Tabulated the data
Volume of standard solution, mL (Vs) Absorbance (As)
0 0.240
5 0.437
10 0.621
15 0.809
20 1.009
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Exercise
Q1:
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Q2:
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Molecular Spectroscopy
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Molecular Fluorescence
Spectroscopy
Fluorescence process in which atoms or molecules
are excited by absorption of electromagnetic radiation.
Then the excited species relax to the ground state,
giving up their excess energy as photons.
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Block Diagram of Molecular Fluorescence Spectroscopy
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Relaxation Processes
Once the molecule is excited to higher energy level,
several processes can occur that cause the molecule
to lost its excess energy .
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Energy Level Diagram: Relaxation Process
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Nonradiative Relaxation
Vibration Relaxation:
- involves transfer of the excess energy of a
vibrationally excited species
- take place in less than 10-15 s and leaves the
molecules in the lowest vibrational state of an
electronic excited state.
Internal Conversion:
- involves transfer of the excess energy of a species in
the lowest vibrational level of an excited electronic state
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Fluorescence Emission
Usually observed from the lowest lying excited electronic
state E1 to the ground state E0 because of the internal
conversion and vibration relaxation processes are very rapid
compared with fluorescence.
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Relationship between Excitation Spectra and
Fluorescence Spectra
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Fluorescence and Structure
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The Effect of Structural Rigidity
Fluorescence is particular favored in rigid molecules.
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Temperature and Solvent Effect
The quantum efficiency decreases with increasing
temperature.
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Topics under Molecular
Spectroscopy
Fourier Transform Infrared (FTIR)
Organic UV-Vis Absorption
Effect of Solvent
Inorganic UV-Vis Absorption
Qualitative Analysis
Quantitative Analysis
Molecular Fluorescence
Equipment for Molecular Spectroscopy
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Equipment for Molecular Spectroscopy
UV/Visible spectroscopy
Infrared spectroscopy
Fluorescence spectroscopy
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UV/Visible Instrumentation
Molecular absorption spectrometry
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UV-Vis: Single Beam Instrument
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UV-Vis: Double Beam Instrument
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Single beam UV/Vis Spectrophotometer
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Double beam UV/Vis Spectrophotometer
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Infrared (IR) Instrumentation
Molecular absorption spectroscopy
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Block diagram of FT-IR spectrometer
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Fourier Transform Infrared (FTIR)
FTIR spectrometer can be single beam or double
beam instrument.
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FTIR Spectrometer
Single Beam
Double Beam
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Fluorescence Instrumentation
Molecular fluorescence spectroscopy
General operation:
i. The light emitted by primary source initially passes
through the excitation monochromator which allows a
narrow band of wavelengths to be selected.
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Fluorometer
Both wavelength
Both wavelength selectors
selectors
are filters are
filters
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Spectrofluorometer
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Fluorescence Spectrophotometer
(fluorometer)
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Fluorescence Spectrometer
(spectrofluorometer)
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END OF MOLECULAR
SPECTROSCOPY
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