odule 6: Spectroscopic, Diffraction, and Microscopic
M
Techniques
(I) Fundamental Concepts in Spectroscopic and Instrumental Techniques
● Spectroscopy Basics
○ Spectroscopy is the study of the interaction between electromagnetic (EM)
r adiation and matter.
It is a fundamental tool for investigating the structures of atoms and
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molecules.
○ The core principle of all spectroscopic techniques involves irradiating a
sample with EM radiation and analyzing the sample's response as a function
of radiation wavelength.
● Types of EM Radiation Interaction with Matter
○ When EM radiation interacts with a substance, it can undergo several
processes:
■ Absorption:The sample absorbs specific wavelengths of the radiation.
■ Transmission:The radiation passes through the sample.
■ Reflection:The radiation bounces off the sample.
■ Scattering:The radiation is dispersed in various directions.
■ Photoluminescence:The sample absorbs radiation and then re-emits it
(includes fluorescence, phosphorescence, and Raman scattering).
○ The color of an object is determined by the wavelengths of light that are
t ransmitted or reflected; the remaining wavelengths are absorbed. A higher
degree of absorption corresponds to a darker color.
The interaction between EM radiation and matter is a quantum phenomenon,
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governed by the properties of the radiation and the structural characteristics
of the sample.
○ Spectrochemical methods specifically measure the absorbed radiation to gain
insights into the sample's composition and structure.
(II) Principle and Applications of UV-Visible Spectroscopy Technique
● Principle
○ Molecules absorb specific wavelengths of radiation, and this absorption is
dictated by their molecular structure.
○ The absorption spectrum reveals absorption bands that correspond to
s pecific structural (functional) groups within the molecule. For instance, the
carbonyl group (=CO) in different molecules (e.g., acetone and diethyl ketone)
absorbs at a similar wavelength.
UV-Vis spectroscopy involves the absorption of energy by a molecule in the
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ultraviolet (UV) region (1-400 nm) or the visible region (400-750 nm). This
absorption results in the excitation of valence electrons to higher energy
levels.
● Beer-Lambert Law
○ This law quantifies the relationship between light absorption and the
roperties of the absorbing medium.
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The absorption of light passing through a material is directly proportional to
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both the distance the light travels through the sample and the concentration
of the absorbing substance.
○ Equation: A = εcl
■ A = Absorbance (no units)
■ ε = Molar absorptivity coefficient (L mol-1 cm-1). A measure of how
s trongly a substance absorbs light at a particular wavelength.
c = Concentration of the analyte (mol L-1)
■
■ l = Path length of the cuvette (cm)
○ Chromophore:Any isolated covalently bonded group in a molecule that is
r esponsible for absorption in the UV-Vis region. Commonly, these are systems
with π electrons and heteroatoms with non-bonding electron pairs.
○ Auxochrome:A group of atoms attached to a chromophore that modifies the
hromophore's ability to absorb light (i.e., alters the wavelength and/or
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intensity of absorption). Examples include -COOH, -OH, and -NH2.
● Electronic Transitions in UV-Visible Spectroscopy
○ Electronic transitions occur when electrons in a molecule are excited from
ne energy level to another by the absorption of UV-Vis radiation.
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Types of electronic transitions:
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■ σ to σ* transitions: High-energy transitions involving the excitation of
lectrons from a bonding σ orbital to an antibonding σ* orbital.
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n to σ* transitions: Transitions for saturated compounds containing atoms
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with lone pairs (n electrons). These transitions require less energy than σ
to σ* transitions and are typically initiated by light in the 150-250 nm
range.
■ n to π* and π to π* transitions: These transitions occur in molecules
containing unsaturated groups (double or triple bonds). They are the basis
of most UV-Vis spectroscopy of organic compounds and fall in the
200-700 nm spectral region.
○ Spectral Shifts:
■ Bathochromic shift (red shift):A shift of the absorptionmaximum to a
longer wavelength.
Hypsochromic shift (blue shift):A shift of the absorptionmaximum to a
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shorter wavelength.
■ Hyperchromism:An increase in molar absorptivity (increased absorption
intensity).
■ Hypochromism:A decrease in molar absorptivity (decreased absorption
intensity).
(III) Principle and Applications of X-Ray Diffraction (XRD) Technique
● Principle
○ X-Ray Diffraction (XRD) is a technique used to determine the crystallographic
s tructure of materials.
○ The technique involves irradiating a material with incident X-rays and
measuring the intensities and angles at which the X-rays are diffracted
(scattered) by the material.
○ XRD is a versatile and non-destructive technique for identifying unknown
crystalline materials.
○ It is also employed to study the structure and function of biological molecules,
including vitamins, drugs, proteins, and nucleic acids (e.g., DNA).
○ Additional applications of XRD include determining structural properties such
as lattice parameters, strain, grain size, epitaxy, phase composition,
orientation, and atomic arrangement, as well as measuring film thickness.
○ XRD can also provide information on how the actual structure of a material
deviates from the ideal structure, revealing internal stresses and defects.
● How XRD Pattern is Produced: Bragg Model of Diffraction
○ Crystalline materials have a regular, periodic arrangement of atoms, while
-rays are a form of EM radiation with wave-like properties.
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When X-rays interact with a crystal, the X-rays are scattered by the electrons
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of the atoms in the crystal (elastic scattering).
○ The regular arrangement of atoms in a crystal acts as a periodic array of
scattering centers, producing a regular pattern of spherical waves.
○ In most directions, these scattered waves cancel each other out through
destructive interference. However, in certain specific directions, the waves
combine constructively, resulting in diffraction peaks.
○ Bragg's Lawdescribes the conditions for constructive interference:
■ nλ = 2dsinθ
■ n = integer representing the order of diffraction
■ λ = wavelength of the X-rays
■ d = spacing between the diffracting planes in the crystal lattice
■ θ = angle of incidence of the X-ray beam with respect to the crystal
planes
● What is Diffraction?
○ Diffraction is the phenomenon where a wave bends or spreads out when it
encounters an obstacle or passes through a narrow opening.
● Interference of Diffracted Waves
○ Interference occurs when two or more diffracted waves interact with each
other.
○ Constructive Interference:Occurs when the diffracted waves are in phase
( their crests and troughs align). This results in a resultant wave with a higher
amplitude. For constructive interference, the path difference between the
waves is an integer multiple of the wavelength (nλ).
Destructive Interference:Occurs when the diffracted waves are out of
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phase. This results in a reduction or cancellation of the wave amplitude. For
destructive interference, the path difference is a multiple of (n/2)λ.
● Components of an XRD Instrument
○ X-ray tube:Generates the X-ray beam.
○ Incident-beam optics:Filters and conditions the X-ray beam before it strikes
t he sample.
○ Goniometer:A device that holds and moves the sample, X-ray optics, and
detector.
○ Sample holder:A mount that secures the sample in the path of the X-ray
beam.
○ Receiving-side optics:Conditions the diffracted X-ray beam after it exits the
sample.
○ Detector:Measures the intensity of the diffracted X-rays.
○ Incident angle (ω):The angle between the incident X-ray beam and the
surface of the sample.
○ Diffracted angle (2θ):The angle between the incident X-ray beam and the
diffracted beam.
○ In a typical XRD instrument, the X-ray tube is stationary, while the sample and
detector are rotated to scan a range of angles.
● XRD Patterns
○ Different materials produce unique XRD patterns, which serve as a fingerprint
f or identifying the crystalline phases present.
Amorphous materials(e.g., glass), which lack long-rangeatomic order,
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produce broad, diffuse patterns.
○ Crystalline materialsproduce sharp, well-defineddiffraction peaks, with the
positions and intensities of the peaks related to the crystal structure.
● Scherrer Equation
○ The Scherrer equation is used to estimate the average size of crystallites
( ordered crystalline domains) in a material from the broadening of the
diffraction peaks.
Crystallites smaller than approximately 120 nm exhibit broadening of the
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diffraction peaks.
○ Equation: τ = Kλ / (βcosθ)
■ τ = mean size of the crystalline domains
■ K = dimensionless shape factor (~0.9 , but depends on crystallite shape)
■ λ = X-ray wavelength
■ β = line broadening at half the maximum intensity (FWHM) in radians, after
subtracting any instrumental broadening.
■ θ = Bragg angle
