Applied Chemistry

Module-4:Spectroscopic and diffraction techniques

Syllabus: Fundamental concepts in spectroscopic and diffraction
techniques – Principle, instrumentation, and applications of UV-Visible
and powder XRD determination of crystallite size, numerical

1
 Introductionto Electromagnetic Radiation
Contents
• Definitionof Spectroscopy
• Electromagnetic Radiation
• Propertiesof EMR
• Interaction of EMR withMatter
• Regionsof EMR,Molecular Effects and its applications
• Principle and Classificationof spectroscopy
• Numerical Problems
• Spectroscopic Instrumentsin Space ExplorationMissions
2
 Spectroscopy
What is spectroscopy?
• The interaction of electromagnetic radiant energy withmatter is called
spectroscopy.
• Quantitative and qualitative analysis of inorganic, organic,and biologicalcompounds.
• Spectroscopic datarepresentsresponse
Spectrum Optics asa function of wavele ngth/frequency.

• Sir Isaac Newton introduced word spectrum in 1672 to describe rainbow ofcolors.
• Fraunhofer made significant contributions to optics and spectroscopy in 1814.
• KirchhoffandBunsen demonstrated spectroscopy's utility in chemical analysis in 1859.
• Spectroscopy has significantly advanced in the fields of chemistry, physics, and astronomy
since its discovery. 3
 Electromagnetic Radiation
• Energy is defined asthe capacity to do work.
• Energy Transfer Methods
• Conduction refers to the transfer of heat from one
substance to another through direct contact.
• Convectionrefersto the transfer of heat through a
fluid due to molecular motion.
• Radiationrefersto the energy transmittedthrough
rays, waves, or particles.
• The dualnature of radiationwasproposed by de Broglie in
1923,
• EMR behavesasa wave in space and as aparticle in matter
interaction.
• The wave nature is measured in termsof frequency,
wave-length, and velocity.
• The particle nature is expressed in individual photons,which
are discrete energy packets. 4
 Propertiesof EMR
• Wave length (λ) refersto the distance between the
adjacent crestsor troughsof a wave ( cm).
• The frequency (v) is the number of oscil ations per unit
time, denoted by hertz (Hz).
• The wavenumber-1 (ν) refersto the number of wavesper
unit distance (cm ).
• Velocity (c) is8 the -1speed of propagationin vacuum,
c=2.9979x10 m.s .
• The energy carried by anEMRor photonis directly
proportional to its frequency, asE=hv=hc/λ.
• Wave is the way of transferringanenergy from one place to
another.
• Photons are discrete packets of energy or quanta.
 Interaction of EMR
• Absorption:EMRinteractswithmatter, causing
with Matter
moleculesandatoms to absorb energyand reach a
higher energystate.
• Emission: It occurswhen asubstance emits
electromagnetic radiation.
• Transmission: The processbywhichincident
radiationpassesthroughmatter without measurable
attenuation.
• Reflection: Measuresthe amountof energyreflected
bymatter.
• Scattering: EMR scattering occurs whenradiation • Wave nature explains reflection,
interactswith matter, causing the energyto be refraction, dif fraction.
reradiated in differentdirections, not along the pathof • Lightabsorptionand emission due
the radiation. toparticle nature in individual
• Refraction: Refersto the deflectionorbending of EM photons.
waves astheypass fromone transparentmediumto
another .
These processesformthe basis fordetectionor spectroscopic measurement 6
 Regions of EMR, Molecular Effects and Applications
Region/ units Molecular Wavelength Frequency (Hz) Energy (eV) Spectroscopy Applications
effects (0A)
Radio/ Nuclear spin >109 < 3x109 < 10-5 Molecular identityand
MHz transitions structure.
Microwave Rotational 109-106 3x109 -3x1011 10-5 -0.01 Bond length of calculations.
GHz/THz/cm- motion
1

Infrared/cm-1 Vibrational 106 - 7000 3x1011 -4.3x1014 0.01 -2 Functionalgroup analysis.

Visible/nm Electronic 7000-4000 4 x1014 – 7.5x1014 2-3 Studyof molecules with
transitions conjugated πandn systems.
Ultraviolet/nm Electronic 4000-10 7.5x1014 -3x1016 3-103 Studyof molecules with
transitions conjugated πandn systems.
X-rays/ Ionization 10-0.1 3x1016 - 3x1019 103-105 gives informationabout the
KeV/eV size and shape of molecules.
Gamma ionisation <0.1 >3x1019 > 105 identify variousradioactive
Rays/MeV isotopes
1eV = 96.48kJ/mol
Contd.
 Contd.
Spectroscopy

Electromagne Alpha Beta Protons Acoustic Seismic

tic spectrum particles particles waves waves
Neutrons
Neutron
radiography Neutron-Induced
Neutron gamma spectroscopy
spectroscopy

Radio Microwave Infrared UV-Visibl AAS X-Ray Diffraction Gamma ray

e
 Principle and Classification of Spectroscopy
• Electromagnetic energy is transferred to atoms, ionsor moleculesin the sample
resultsin a transitionto higher energy state.
• Energy required of photonto give this transition.

hν= ∆E = E1-E0

10
 Numerical Problems
1. Whatisthefrequency ofradiationwith a wavelength of5.00 x 10¯8 m?Inwhat
region ofthe electromagnetic spectrumis this radiation?
• Use λν = c to determine the frequency:
• (5.00x 10¯8 m)(ν)= 3.00x 108 m/s = 6.00 x 1015 s¯1
• Frequency in Middle Ultraviolet Region
2. Calculate thewavelength ofradiationemitted from radioactivecobalt witha
frequency of2.80 x1020 s¯1. What region of theeletromagnetic spectrumdoes
thisliein?
• Use λν = c to determine the wavelength:
• λν = c 20 1
• λ (2.80 x 10 s¯ )= 3.00x 108 m.s-1
• λ = 1.07 x 10¯12 m
Gammarays.
11
 Spectroscopic Instrumentsin SpaceExploration Missions
• ExplorationRoversLaunch
• Searching and analyzing rock and soils.
• Utilizing multiple spectrometers for sample analysis.
MarsMission
• Mini-TES: miniature thermal emission
spectrometer (examine rock, soil &
atmosphere)
• MB: Mossbauer Spectrometer (examine
mineralogy of rocks & soils)
• APXS:AlphaParticle X-ray Spectrometer
(analyze elementsin rocks& soils)
Chandrayaan 3 mission
• Seismometer, Langmuir Probe
• Thermal Probe, Retroreflector
• X-ray Spectrometer. Laser-based
Spectrometer
• Spectro-polarimeter
Principle,Instrumentation and Applications ofUV-visible Spectroscopy

13
 Contents
• UV Visible Light AbsorptionRegion
• Absorption and Emission
• Electronic Transitions
• Terms and Shifts used in UV Spectroscopy
• Color-absorptionrelationship
• Principles ofUV Spectroscopy-Absorption Law
• Instrumentation
• Applications
• Numerical Problems
• Summary
14
 UV VisibleLightAbsorption Region
• The processinvolvesthe absorption of light from 200 nm-700 nm by moleculesor
atoms.
• UVand visible light is involved withelectronic excitation.
• UV-Vis radiationhasthe potentialto excite valence electrons in moleculesor atoms.

• Near UV/UVA: Long-waveUV or blacklight, not absorbed by O3

layer
• MediumUV/UVB: medium-waveUV, mostly absorbed by O3
layer 15
• Far UV/UVC: short-waveUV, ionizingradiation, absorbed by O3
 Absorption and Emission
• leAbsorpti
vels. onand emission processes in volv e transitions betweendifferent energy
• Incidence photonmust equal energy difference between states;hν= ∆S= S1-S0.
• Energy absorptionallows transitionto excited state.
• The energy of molecular orbitals beingspecific to eachgroup or chemical function.
• Absorptionwavelengthis thus correlated to the type of chemical bonds.

16
 Electronic transitionsand Absorption Bands Intensity
• Afterexcitationby EMR, valence electrons take part in electronic transitions.
• Four different transitionsare allowed betweenσ, π and lone pair electrons.
1. Transition σ →σ *
• occurs only for simple bonds(C-H or C-C for example)
• Requireshigh energyand found in 100nm-150nm
2. Transition n →σ *
• occurs for atoms withlone pair electrons (N, O, S….)
• requires ahighquantity of energy, and is thusfound
between 150 and *250nm.
3. Transition n→ π
• Only occurs for atoms withlone pair electrons and
unsaturated functi*ons.(250nm- 500nm)
4. Transitionπ →π (aromatics)
• occurs only for atoms withunsaturatedfunctions • σ →σ*>n →σ*>π →π*>n→ π*
• are locatedin the UVand visible domain (200to 700 nm) • Forbiddentransition: ε ≤ 10
• Absorptionbandsintensity linked to electronic • Partlyallowed transition: 10 ≤ ε≤
transitionprobability. •
1000
Allowed transition:1000 ≤ ε ≤
• Molar absorptioncoefficient valid forone wavelength. 100000 17
• Highly allowed transition:ε≥ 100000
 Terms involved in UV Spectroscopy
• A chromophore is any isolated covalently unsaturated bonded group that shows a
characteristic absorption of EMR in the UVor Visible region.
¸ Organic molecules containing >C=C<, >C=O, -N=N- and -NO2
In Greek: Chroma = color, and phoros= bearer.
• Anauxochrome is group of atomsattached to a chrmophore which modifies the
¸

ability of that chromophore to absorblight.

¸ Organic molecules containing:- COOH, -OH,-SO3H, -NR2 -NH2
In Greek, Auxo= increase and chrome=color.
• Chromgenis the chrmophore and auxochromo together.
¸
 Shifts Involved in UV Spectroscopy
• Bathochromic Shift ( Red Shift)
oShiftsabsorptionmaximumtowardslonger
wavelength.
oCaused by auxochrome presence or solvent change.
• Hypsochromic Shift ( Blue Shift)
oShiftsabsorptionmaximumtowardsshorter
wavelength.
oCaused by removal of conjugation and solvent
polarity.
• Hyperchromic shift
o Increasesabsorptionmaximum intensity.
oAuxochrome usually boostsabsorptionintensity.
o B-band shifts from 275 mμ to 262mμ.
o Emax for 2-methyl pyridine increases to 3560.
• Hypochromic Shift
oDecreasesabsorptionmaximum intensity.
 Color-absorption Relationship
• Colored solution absorbs light corresponding to complementary color that is opposite
color in chromatic circle.
• Methylene blue absorbs600-700 nm in organe-red, so that it looksblue for us
• Cong red absorbs maximum at 500 nm in green, so that it looks like red for us.
• Red and blue light are absorbed by chlorophyll and so it appearsitscomplementary
color, green.
0.75 Methylene
ecnabrosbA blue
0.5 Congo red
0.25
0
400 500 600 700 800
Wavelength
(nm)
 Principles of UV Spectroscopy-Absorption Law
• Beer–Lambert law orBeer–Lambert–Bouguer law.
• The law combinesdiscoveriesmade by Bouger, Lambert,and Beer (1729-1852)
• At a wavelengthmaximum, absorption is proportional to the concentrationof
absorbing molecules and the path lengthof the light throughthe sample.
• A = εLC
• A = absorbance of the sample, log10 (I0 /I)
• I0 = intensity of the radiation entering the sample
• l = intensity of the radiation emerging from the
sample
• L = length of the light path through the sample, in
centimeters
• C = concentration ofthe sample,-1 in mol e s/lit er
• ε = molar absorptivity (Litre mol cm-1 )
• It appliesto all formsof electromagnetic radiation, not just visible
light. 21
 Instrumentation
• Arnold O. Beckman's created Spectrophotometry in
1940

• A "single-beam" spectrophotometer is
made up of:
• a polychromatic source
• a wavelengthselector and eventually a
source selector
• a sample compartment
• a detector
• a screen and/or acomputer.
22
Instrumentation-Double-beamspectrophotometer

23
 Applications
1. Quantitativeanalysisof analyte: Determinationof the concentrationof a
compound in solutionusing Beer’s law.
Determinationofpercent content of Vitamin A1 and Vitamin A2 in natural fatsandoils by
measurementof intensities of peaks at λmax=325nm and 351 nm respectively.
2. Distinctioninconjugated and non-conjugated compounds: if an organic
compound doesnot show λmax above 200 nm cannot contain conjugated
chromophore, aldehyde,keto group, benzene ring.
3. Distinctionbetween Cis and Trans geometricalisomerism: Transisomer shows
longer absorptionwavelengthand Higher intensities thancis isomer.
4. Distinctionbetween keto-enol tautomerism: Keto form showslow intensity band
at 275 nm (ε=20) due to isolated keto-carbonyl group while enol showshigh
intensity band at 245 nm (ε=1800) due to conjugated double bond.
5. Study of charge transfer complex: Charge transfer complexesare formed when
a electronrich molecules (teracynaoethylene iodine) is mixed with and electron
acceptor molecules( aniline,benzene). The charge transfer complex is
responsible for new –absorption peaks. 24
 Numerical Problems
1. Asample-1 hasa maximumabsorbance of275 nmand molar absorptivity of
8400 M cm-1. Aspectrophotometer measures absorbanceof 0.70 using a
cuvettethat is1 cmwide. Find the solution concentration.
Use the equation A= εLcand and solve for concentration(c):
• c = A/ε
• c = (0.70)/(8400 M-1cm-1)(1 cm) = 8.33 x10-5 mol/L = 8.33 x10-5 M
2. Indicate thefunctional groupsshown in thecompound below whether they are
auxochromeor chromophore groups

3. What sort ofelectronic transitionsare possibleforeach ofthe following

compounds? (a) cyclopetane(b) acetaldhyde (c) dimethyl ether (d) triethyl
amine(e) cyclohexene 25
Principle, Instrumentation and Applications of XRD

26
 Contents
• X-rays and Energy.
• Continuous and Characteristics X–rays.
• Production of X-rays.
• Diffraction and Interference.
• Principles ofX-ray Diffraction(XRD).
• Bragg’s Law.
• X-ray Diffractometer.
• X-ray Diffraction Pattern Analysis.
• Crystallite Size from XRD.
• Numerical problems.
• Applications,Advantages and
Disadvantages. 27
 X-rays and Energy
• It is basically energetic high-frequency • Discovered by [Link] 1895.
EMR. • NobelPrize in 1901.
• Part of electromagnetic spectrum: • X-radiation (X-rays)or Röntgen
o Wavelength:0.01 -10 nm shorter than UV
and grater than gamma.
radiation.
o Energy: 1keV-100 keV higher than UV and • Types of X-rays:HardX-rays(0.01 to 1
lower than gamma. nm) and Soft X-rays(0.1 to 10 nm).
o Frequency: 3x10 Hz -3x10 Hz.
16 19 • Analytical purpose: 0.7 to 2nm
• First example of ionizing radiation.

• Anna Bertha Ludwig, wife of Rontgen

• First humanbodyradiographic image
28
 Continuous and Characteristics X-rays
 Continuous (Bremsstrahlung, breakingradiation)
X-rays
• Nucleus field interaction:Bremsstahlung
• Arises from electron deceleration.
• Electrons near nucleus slow down KE.
• Loss of KE compensated by X-ray emission.
• Polychromatic, continuous, or white radiation.
• Phenomenon was
 Characteristic X-rays
discovered by Nik ola Tesla.
• Inner shell interaction produces characteristic radiation.
• Created when ahole in inner shell fil ed byhigher energy
electron.
• K-shell electronknockedoutfil s vacancybyL-shell (Kα)or
M-shell (Kβ) electron.
• K-linesare mostuseful in X-ray diffraction.
. • CuKα is commonly used onlab scale x-rayinstruments:
• Energyis 8.04keV, corresponding to1.5406 Å X-ray λ.
• Used for the crystal structure studyby XRD.
• Discovered byCharles Glover Barkla in 1909. 29
 Production of X-rays
• Evacuated glass/metal bulb.
• Seals cathode and anode electrodesoneachend of the tube.
o Cathode (-ve):Tungsten (W) ( Filament type).
o Anode ( +ve) : Cu/ Cr/Mo/W/ depending type of applications (disktype).
• Electrons are produced by the heatedcathode via thermionic process to hit the anode.
• Accelerating potential is applied betweenthe cathode and anode (25-150 kV).
• X-rays are produced whena fast moving stream of electronsis rapidly decelerated.
• CharacteristicsKα, (Å) of differentanodes: Cr(2.29), Fe (1.94), Co(1.79), Cu (1.54), Mo (0.71) and W (2.13).
• Tungsten features:
o Highatomic numberof74.
o Efficient electronrelease.
o Good thermionic emitter.
o Manufactured as thin
wires.
o HighM.P. of 3422 0C.
Recent model Old modelCrookes tube 30
 Diffraction and Interference
• Interference is the interactionof the two waves:
 Reinforcing interaction = Constructive interference
 Two waves are in phase, results in greater amplitude wave
 Canceling interaction = Destructive interference
 Two waves are out ofphase, cancel each other then no net motion remain

• Bending of light around the

corners of doorandwindows.
• Diffractionoccurs when alight wave passes through aphysical slit or corner
that is smaller than or equalto itswavelength. 31
 Principlesof X-ray Diffraction (XRD)
 Microstructure of solid materials
• Crystalline: Entire solid is made of upatoms in an
orderly array.
• Polycrystalline:Completely ordered in segments.
• Amorphous:No recognizable long range order.
• “Every crystalline substance gives a pattern;the same
substance always gives the same pattern;and in a
mixture of substances each produces it’s independently
of others”.
• The XRD pattern of pure substances resembles a
fingerprint ofthe substance.
• It is based on the scatteringof X –rays by crystals.
• The atomicplanes of crystal cause an incident beam of
X-rays to interfere with one another as they leave the
crystalis described by the famous law called Braggs
• Chemic al formula of SiO
law. • Crystalstructures vary.
2
32
 Bragg’s Law
• The atomic planesof a crystal cause anincident
beam of X-rays to interfere withone another as
they leave the crystal is called X-ray diffraction.
• Two parallelrays create angle ϴ with crystalplanes.
• When the totalpath difference is 2dsin θ equals to nλ,
diffraction occurs:
 Pathdifference (δ)=CB+ BD
 For Δ ACB; Sin θ=CB/AB=CB/d=CB=dSin θ
 Similarly, Δ ADB; Sin θ=BD/AB=BD/d=BD=dSin θ
 So, dSin θ+ dSin θ=2dSin θ
Bragg’s law: δ=2dSin θ=nλ
Where


δ : Pathdifference betweenthe reflected x-raywaves.

λ:IncidentX-raywavelength.
n= aninteger ( 1,2,3…). Nobel Prizes in Physics:
d=distance between lattice planes. • Maxvon Laue: 1914 NobelPrize for X-ray
diffraction discovery.
Θ:angle betweenthe indent X-rayand the lattice. • [Link] andW.L. Bragg: 1915 Nobel prizes
11/23/2025
for X-raycrystal structure analysis. 33
 X-Ray Diffractometer
• It is the instrument used to
produce monochromatic x-rays.
• Focuses beam on sample.
• Scans througha 2θ range.
• Detectsreflected x-rays and their
intensity.
• Attached to a goniometer for
simultaneousscanning.
• Optics focusx-ray beam on
sample.
• Reflects x-raysinto detector.
• Detector: Gas-fil ed or scintil ation
or semiconductor.
• Records peakpositionand
intensity during constructive
interference.
• Displayed ona diffractogram. 34
 X-ray Diffraction Pattern Parameters and Analysis
 XRD Plot: Signalintensity at two thetapositions.
 PeakIntensity and Peakposition
• Two theta positions atoms within a lattice structure.
• iIntensit
n phase. y of peaks correl ates with number of atoms
• mol
Greater peak intensit y indicates
ecules with specific spacing. more crystals or
 PeakWidth and CrystalSize
• Peak width inversely proportional tocrystal size.
• Thinner peak indicates larger crystal.
• amorphous
Broader peaknature.
suggests smaller crystal, defect, or
 Diffraction Pattern Database Confirmation
• Matchesunknowncompound patternswith literature and
experimental values.
• Verifies elementidentity by matching location,width,and Joint Committee on PowderDiffraction Standards
relative heights ofdiffractionpatterns. (JCPDS)
International Crystal Structure Database(ICSD)
35
 Crystallite sizefromXRD
 Crystallite:
• Small crystalcomposed of multiple atoms/molecules.
• A crystallite is that portion of the crystal which diffracts
light coherently.
• Calculated using XRD diffractogram using Scherrer's
formula.
 Grain:
• During crystal growth, multiple nucleation centers
responsible for grain formation.
• Grain contains multiple crystallites.
• Grain size is detected using SEM.
 Particle:
• Consists ofsingle crystal or agglomeration of several
grains.
• Grains combine toform particles.
• Particle size always larger than crystallite size. Particle ≤ grain ≤ crystalli te < unit cell
• Particle size is detected using TEM. 36
 Crystallite Size
• Crystallite size: D= k λ/β cos θ.
• β=FWHM in radians, θ= Bragg angle, λ= X-ray
wavelength.
• k=Scherrer constantdepends on shape.
• 0.94 For FWHM of sphericalcrystals with cubic
symmetry.

P.H.
Scherrer
(1890-1969)
37
 XRD numerical problems
1. If X-rays of wavelength 0.9 Åare diffracted at an angleof7 0 inthe first order, what
is thespacing betweenthe adjacent planes of thecrystal inÅ?
 According Bragg’s law
Solution: ¸ 2dsin θ= nλ
• Wavelength ofX-rays= 0.9 Å = 0.9x10-10 2 x d x sin(70)= 1x 0.9x10-10-10
¸
m ¸ 2x d x 0.1218= 1x 0.9x10
• Angle (θ)= 70 ¸ d= 0.9x10-10/0.2436
• First order reflection=n=1 ¸ d=3.6946x10-10 cm
• Spacing between the adjacent ¸ 3.6946 Å
planes=d=?
2. Estimate the crystallite size of the given nanomaterial usingp-XRD data: Peak position
2θ = 21.61o,FWHM of sample = 2.51o, k = 0.9 and λ = 1.5406 Å(degree toradian = Degree
× π/180). • According to Scherrer’s equation D= :kxλ
Given: o βx Cosθ
¸ 2θ = 21.61 (θ = 10.805o)
¸ β= FWHM = 2.51o (0.043825 radian) • Crystallite size= 0.94x0.15406
¸ k = 0.94 0.043825x0.982257
¸ λ = 1.5406 Å (0.15406 nm) D = 3.22 nm
38
 Applications,Advantages and Disadvantages
 Applications:
• Distinguishing between crystalline and amorphousmaterials.
• Details unit cell dimensions, atom positions, and crystalsymmetry.
• Determining texture of polygrainedmaterials.
• Measuring strain and small grain size.
 Advantages:
• Lowest cost,convenience,and widely used for crystalstructure
. determination.
• No significant air absorption, requiring no evacuated chamber.
• Nondestructive technique
 Disadvantages:
• X-rays do not interact very strongly withlighter elements
39