Module V

Modern Spectroscopic Methods in Chemical Analysis

Topics:
Types of Electromagnetic spectrum, Absorption of radiation: Franck-Condon Principle, Beer-Lambert’s law,
UV-Visible Spectroscopy: types of electronic transitions and Applications, IR spectroscopy: Fundamental
modes of molecular vibrations, Functional group region, finger print region, Applications of IR- Spectroscopy,
NMR spectroscopy-Basic Principle, Instrumentation, Chemical Shift, Applications

5.0 Introduction:
Spectroscopy is the study of how matter interacts with electromagnetic radiation (light, X-rays,
microwaves, etc.). Each substance absorbs different parts of the spectrum in a unique way. If the matter is a
molecule, specifically the term molecular spectroscopy is used.

Using spectroscopy we can identify molecular structure, detect functional groups, study chemical
changes, determine physical and chemical properties. Spectroscopic techniques are fast, accurate and
requires very small sample.
5.1 Types of electromagnetic spectrum:
Electromagnetic spectrum:
Electromagnetic spectrum includes all types of electromagnetic radiations in increasing order of their
wavelengths.
γ-rays < X-rays < U.V. rays < Visible rays < I.R. rays < Microwaves < Radio waves

Electromagnetic Spectrum and Spectroscopy

Wavelength
Region Frequency Range Molecular Process Applications
Range
Gamma Rays < 0.01 nm > 10¹⁹ Hz Nuclear transitions Nuclear decay, astrophysics
Inner-shell electron X-ray crystallography, medical
X-Rays 0.01 – 10 nm 10¹⁷ – 10¹⁹ Hz
transitions imaging
Ultraviolet Valence electron Electronic spectroscopy (UV-
10 – 400 nm 10¹⁵ – 10¹⁷ Hz
(UV) excitation Vis), photochemistry
~4×10¹⁴ – 7.5×10¹⁴ Valence electron
Visible 400 – 700 nm Color, UV-Vis spectroscopy
Hz excitation
Vibrational IR spectroscopy, molecular
Infrared (IR) 700 nm – 1 mm 3×10¹¹ – 4×10¹⁴ Hz
transitions identification
Rotational Microwave spectroscopy,
Microwave 1 mm – 1 m 3×10⁸ – 3×10¹¹ Hz
transitions RADAR
Radiofrequency Nuclear spin
>1m < 3×10⁸ Hz NMR spectroscopy, MRI
(RF) transitions

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5.2 Absorption of radiation: Franck-Condon Principle:
The Franck–Condon principle states that during an electronic transition, such as the absorption or
emission of light in molecules, the positions of the nuclei remain unchanged because electronic transitions
occur much faster than nuclear motion.

Internuclear distances are equal in upper and lower states

5.3 Beer-Lambert’s law:
When a beam of monochromatic light passes through a homogeneous absorbing medium, its intensity
decreases exponentially with both the path length of the medium and the concentration of the absorbing
species.

𝐼𝑡 = 𝐼0 𝑒 −𝑘1 𝑘2𝑙𝑐

𝐼𝑡 = 𝐼0 𝑒 −𝜀𝑙𝑐

where ε (ε =k1k2) is called molar absorptivity (or molar extinction coefficient).

We can also write
𝐼0
log10 = 𝜀𝑐𝑙
𝐼𝑡
𝐼0
log10 = 𝑨
𝐼𝑡
where A = εcl

where this ‘A’ is called Absorbance or Optical density.

Units: ε = dm3mol-1cm-1; A = dimensionless; c = mol dm-3; l =cm

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Graph:

From Beer Lambert’s law equation, A = εcl , if we draw a graph by taking absorbance on y-axis and
concentration on x-axis, we find a straight line passing through origin.

Limitations of Beer-Lambert’s Law:

1. The law does not hold well for highly concentrated solutions
2. It cannot be applied to infinitely dilute solutions
3. The law is not applicable to solutions that are sensitive to pH
4. The Beer–Lambert law is valid only when monochromatic light (light of a single wavelength) is
used.

5.4 UV-Visible Spectroscopy:

UV–Visible spectroscopy is an analytical technique that measures the amount of ultraviolet and
visible light absorbed or transmitted by a sample.
Types of electronic transitions
The following types of electronic transitions are possible.
1. σ → σ* Transitions
• Sigma-bonded electrons are tightly held because of strong head-on overlap.
• To excite them, very high energy (short wavelength) radiation is needed.
• These transitions usually occur below 200 nm (vacuum UV region).
• Example: Alkanes and saturated hydrocarbons such as methane and propane.
2. n → σ* Transitions
• Molecules containing heteroatoms like oxygen, nitrogen, sulphur, or halogens have non-bonding
electrons.
• These can be excited to an antibonding σ* orbital.
• Transitions generally occur in the range of 150–250 nm.
• Example: Alcohols, alkyl halides, and amines.
3. π → π* Transitions
• Found in molecules with double or triple bonds where electrons from a bonding π orbital are
excited to an antibonding π* orbital.
• These transitions occur typically between 200–400 nm.
• Example: Alkenes and aromatic compounds such as ethene and benzene.
4. n → π* Transitions
• Non-bonding electrons on heteroatoms can be excited to antibonding π* orbitals.
• These transitions occur at 250–600 nm, often falling within the visible region.
• Example: Carbonyl compounds such as acetaldehyde and acetone, nitro compounds, and azo
compounds.

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 The relative energies required for the various transitions follow the order.
σ→σ* > n→σ* >π→π* >n→π*

Applications of UV–Visible Spectroscopy

1. UV–Visible spectroscopy is used for identifying compounds based on their characteristic absorption
bands.
2. It is used to differentiate cis–trans isomers through differences in their absorption wavelengths.
3. It is used to detect the presence and extent of conjugation in molecules.
4. It is used to detect impurities that absorb in the UV–Visible region.
5. It is used to study hydrogen bonding through shifts in absorption maxima.
6. It is used to determine dissociation constants (pKa) by measuring absorbance at different pH values.
7. It is used to study reaction kinetics by monitoring changes in absorbance with time.
8. It is used for quantitative analysis of compounds using Beer–Lambert’s law.

5.5 IR Spectroscopy:
In the electromagnetic spectrum, the region extending from about 12,500 cm⁻¹ to 50 cm⁻¹ is known
as the infrared (IR) region This region is divided into:
Near IR: 12,500–4000 cm⁻¹
Mid IR: 4000–400 cm⁻¹
Far IR: 400–10 cm⁻¹

• Near-IR shows overtone and combination bands.

• Mid-IR shows fundamental vibrations, and most functional groups absorb in this region; it also
contains the functional group and fingerprint regions.
• Far-IR shows lattice vibrations and heavy-atom bond vibrations, useful for inorganic and solid-state
studies.

The infrared spectrum is usually studied under two important regions:

Functional group region (4000–1500 cm⁻¹):
Shows characteristic absorption peaks of functional groups such as O–H, N–H, C–H, C=O, and
C=C. It is also called the diagnostic region.

Fingerprint region (1500–667 cm⁻¹):

Shows a complex pattern of absorptions due to combined vibrations of the whole molecule. This
region is unique for every compound, just like a fingerprint, and is used for molecular identification
by comparison with reference spectra.

Different molecules absorb different IR frequencies. This property is used to determine the structure
of compounds. Modern IR spectroscopy mainly uses FTIR (Fourier Transform Infrared) instruments, which
provide rapid, sensitive, and accurate results.
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Modes of vibrations:
A molecule may be viewed as a system of balls (atoms) connected by springs (bonds). When it
absorbs IR radiation, the bonds undergo deformation, leading to stretching and bending vibrations.

I. Stretching vibrations: In this type of vibrations, the distance between the atoms increases or decreases
but the atoms remain in the same bond axis. Stretching can be of two types.
a) Symmetric: The movement of atoms with respect to a particular atom is in the same direction.

Symmetric stretching
b) Asymmetric: One atom approaches the central atom and the other departs from it.

Asymmetric stretching vibrations

II. Bending vibrations: In these types of vibrations, the position of the atom changes with respect to the
original bond axis but the distance between the atoms remains constant.
Bending vibrations are of two types
a) In-plane bending: Atoms remain in the same plane.

b) Out of plane bending: Atoms move out of the plane with respect to the plane of the central atom.

Wagging: Both atoms swing up and Twisting: One atom moves up and
down the other moves down the plane

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Applications of IR Spectroscopy

1. IR spectroscopy is used to find the basic molecular structure of compounds.

2. It is used to identify the functional groups present in molecules.
3. It is used to analyze samples in solid, liquid, and gaseous states.
4. It is used to record spectra through absorption, emission, and reflection methods.
5. It is used in hydrogen-bonding studies, impurity identification, and quality control.
6. Used to distinguish between different isomers and molecular conformations.
7. Used to study physical properties such as phase changes, solubility, diffusion at the molecular level

5.6 NMR Spectroscopy:

Basic Principle
In NMR spectroscopy, nuclei with magnetic moments align either parallel or antiparallel to a strong
external magnetic field. When radiofrequency radiation of the appropriate energy is applied, these nuclei
absorb the energy and undergo a transition between spin states. As they relax back to their lower-energy state,
they emit signals that provide information about the molecular structure and the chemical environment of the
atoms.

• NMR-active nuclei are those with a nuclear spin quantum number I ≠ 0 (i.e., with at least one
odd proton or odd neutron). Examples: ¹H, ¹³C, ¹⁹F, ³¹P (all have I = ½).
• NMR-inactive nuclei have I = 0, so they do not produce NMR signals (e.g., ¹²C, ¹⁶O, ³²S).

Instrumentation and Working of NMR Spectrometer:

A modern NMR spectrometer arrangement is as shown in the following figure.

1. Magnet – Provides a strong and stable magnetic field. Modern instruments use superconducting
magnets cooled by liquid helium/nitrogen.
2. Radiofrequency (RF) Source – Produces short pulses of RF radiation in the MHz range (e.g., 60
MHz, 100 MHz, 400 MHz instruments).
3. Sample Probe & Coil – The sample (in a deuterated solvent such as CDCl₃, D₂O, etc.) is placed in a
thin glass tube. The probe and coil transmit RF pulses and detect the emitted signal.
4. Detector / Receiver – Detects the signals emitted when nuclei relax back to their lower energy states.

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 5. Computer & Data System – In Fourier Transform NMR (FT-NMR), pulsed RF radiation is used. The
computer performs a Fourier transform to convert time-domain data into a frequency-domain
spectrum.

Working of the Instrument:

1. The sample is placed in the magnetic field (B₀). The nuclei align with or against the field.
2. A short RF pulse is applied, causing nuclei to move into resonance.
3. When the pulse stops, nuclei relax back, emitting a Free Induction Decay (FID) signal.
4. The FID is detected by the receiver coil.
5. The computer applies a Fourier Transform to convert the time-domain signal into a frequency-
domain spectrum.
6. The spectrum shows chemical shifts, which can be analyzed for structure determination.
Chemical Shift (δ):
Chemical shift is the difference in resonance frequency of a nucleus compared to a reference (usually
tetramethylsilane, TMS) under the same conditions.
ν𝑠𝑎𝑚𝑝𝑙𝑒 − 𝑣𝑟𝑒𝑓𝑒𝑟𝑒𝑛𝑐𝑒
𝐶ℎ𝑒𝑚𝑖𝑐𝑎𝑙 𝑠ℎ𝑖𝑓𝑡 (δ) = 𝑋106 𝑝𝑝𝑚
ν𝑟𝑒𝑓𝑒𝑟𝑒𝑛𝑐𝑒
TMS is set at 0 ppm. Proton resonances usually appear in the range 0–15 ppm. Electrons around a
nucleus induce small magnetic fields.

• If the induced field opposes B₀, the nucleus is shielded and resonates at lower δ (upfield).
• If the induced field adds to B₀, the nucleus is deshielded and resonates at higher δ (downfield).

Type of Proton δ (ppm)

Alkanes (–CH₃, –CH₂–) 0.9–1.5
Alkenes (–CH=) 4.5–6.5
Aromatics 6.5–8.0
Aldehydes (–CHO) 9–10
Carboxylic acids (–COOH) 10–12

Applications of NMR Spectroscopy:

1. NMR spectroscopy is used for determining the structure of molecules.

2. It is used to identify functional groups based on characteristic chemical shift ranges.
3. It is used to study stereochemistry and conformational details of molecules.
4. It is used to investigate hydrogen bonding through shifts in chemical signals.
5. It is used to detect impurities and assess the purity of samples.
6. It is used for quantitative analysis by integrating signal intensities.
7. It is widely used in pharmaceutical research and drug development.