SECTROSCOPIC

METHODS OF ANALYSIS
‫طرق التحليل الطيفي‬
04036411

FALL
2022/2023
Course aim
Discuss theory and background for some methods of spectroscopic analytical
techniques

References
• Analytical Chemistry, Gary D. Christian, 7th Edition (2014)
(Chapters 16 and 17)

• Introduction to Instrumental Analysis, Robert D. Braun (1987)

• Physical Methods in Inorganic Chemistry, Russell S. Drago (1965)
 Course contents
1- Introduction to spectroscopic methods of analysis
2- Absorption theories and laws
3- Spectroscopic Instrumentation: Types and components
4- Electronic Spectroscopy:
– UV-Vis molecular absorption spectroscopy
– Applications of absorption measurement and photometric titration
5- Fluorescence, phosphorescence, and chemiluminescence.
6- Infrared (IR) absorption spectrometry:
– Theory and its applications
– Raman spectroscopy
7- Atomic absorption spectroscopy
8- Atomic emission spectroscopy and ICP method
 Chapter 1
Introduction to spectroscopic
methods of analysis
Instrumental analysis
Instrumental analysis is a field of analytical chemistry that investigates
analytes (samples) using scientific instrument.

Spectroscopic methods: Instrumental analysis II (4th year)

Thermal methods

Mass analyzers

Separation methods: Instrumental analysis I (3rd year)

Chromatography and Electrophoresis

Electrochemical methods:
Measures the voltage and electric current
 Spectroscopic methods of analysis:
❑ Measures the interaction of the sample with electromagnetic radiation (EMR)
❑ There are two types of spectroscopy

Atomic Spectroscopy: Molecular Spectroscopy:

Spectral lines due to electronic Band spectra due to electronic

transitions. Atoms of different transitions, molecular vibrations
elements have distinct spectra and and molecular rotations.
therefore atomic spectroscopy allows Examples
for the identification and quantitation
of a sample's elemental composition UV-Vis Spectroscopy:

Examples
Infrared Spectroscopy:

Atomic Absorption Spectroscopy: Raman Spectroscopy:

Atomic Emission Spectroscopy: Fluorescence Spectroscopy:

 Interaction of EMR with matters:
Depends on the energy of the photons

• Transmission (transparent matter)

• Absorption (Photoionization
Electronic transition
molecular vibration
molecular rotation)

• Classical interactions
• Reflection
• Refraction
• Scattering
• Polarization
 Electromagnetic Radiation, EMR:
Frequency, n in Hz
~1019 ~1017 ~1015 ~1013 ~1010 ~105

Wavelength, l
~.0001 nm ~0.01 nm 10 nm 1000 nm 0.01 cm 100 m

Energy (kcal/mol)
> 300 300-30 300-30 ~10-4 ~10-6

g-rays X-rays UV IR Microwave Radio

nuclear core electronic molecular molecular NMR and MRI
excitation electron excitation vibration rotation
(PET) excitation (p to p*)
(X-ray
cryst.)

Visible
 Positron-emission tomography (PET)
• Is a nuclear imaging technique that is used to the
diagnosis of disease.
• The system detects gamma rays emitted by a positron-
emitting radioligand, most commonly fluorine-18,

• 18F is introduced into the body on a biologically

active molecule called a radioactive tracer
(fluorodeoxyglucose (FDG))
• the concentrations of tracer imaged
will indicate the cell metabolic
activity and the glucose uptake.
Magnetic Resonance Imaging (MRI)
• is a medical imaging technique used
in radiology to form pictures of the anatomy of
the body.
• MRI scanners use strong magnetic fields
and radio waves to generate images of the
organs in the body.
Common Spectroscopic methods depends on EMR :
Type of Spectroscopy Usual Type of transition
wavelength
range
Gamma Ray Emission 0.005 - 1.4 Ao Nuclear
X-ray absorption / emission 0.1 – 100 Ao Inner (core) electrons
Vacuum ultraviolet absorption 10 – 200 nm Bonding electrons
UV-Vis absorption, emission and 200 – 780 nm Bonding electrons
fluorescence
Infrared absorption and Raman 0.78- 300 m Rotation / vibration of
molecules
Microwave absorption 0.75 – 375 mm Rotation of molecules
Electron spin resonance 3 cm Spin of electrons in a
magnetic field
Nuclear magnetic resonance 0.6 – 10 m Spin of nuclei in a
magnetic field
 UV-Vis Spectroscopy :

The ultraviolet (UV) region

* extends from about 10 to 380 nm:
* the most analytically useful region is from 200 to 380 nm,
called the near-ultraviolet or quartz region.
* Below 200 nm, air absorbs, so instruments are operated under
vacuum and this region is called vacuum-ultraviolet region or Far UV.

The visible (Vis) region

*can be seen by the human eye, where the light appears as a color.
*It extends from near-ultraviolet region 380 nm to about 780 nm.
 IR region:
• The near IR (NIR): it is the higher-energy near-IR,
lying adjacent to the visible region approximately
12500–4000 cm−1 (0.8–2.5 μm wavelength).

• Mid- IR: approximately 4000–400 cm−1 (2.5–25 μm) is

used to study the fundamental vibrations and
associated rotational-vibrational structure. It is called
also NaCl-IR region

• Far- infrared: approximately 400–40 cm−1 (25-

250 μm), lying adjacent to the microwave region, has
low energy and is used for rotational spectroscopy.
Remember
There are three common types of transitions:
❑ Rotational transition: the molecule rotates about various axis. Molecule absorbs
radiation and is raised to higher rotational energy level.

❑ Vibrational transition: the molecule absorbs a discrete amount of energy and is raised to
higher vibrational energy level.

❑ Electronic transition: the electrons of a molecule may be raised to a higher electron

energy level.

The relative energy levels of the three transition processes are in the order:
electronic > vibrational > rotational.
Radiation Transition Type Shape of Spectra
------------------------------------------------------------------------------------------------------
Microwave Rotational Sharp lines
Infrared Vibrational / Rotational Peaks/bands
Visible-UV Electronic / Vibrational / Rotational Broad bands
(Outer electrons electronic transitions)
------------------------------------------------------------------------------------------------------
Give Reason:
The electronic transitions appear as a broad band.
Because each electronic energy level has multiple vibrational and
rotational energy levels associated with it. Vibrational energy
levels are very close in energy, rotational energy levels are even
closer.
 Light and the observation of colors
When light falls on a substance, and a color is seen, this
means, certain wavelength is absorbed, and the
remainder is transmitted
If a sample absorbs all wavelengths in the visible
region of the spectrum, it will appear black; if it
absorbs none of them, it will appear white or
colorless. • Violet: 400 - 420 nm
• Indigo: 420 - 440 nm
• Blue: 440 - 490 nm
• Green: 490 - 570 nm
• Yellow: 570 - 585 nm
• Orange: 585 - 620 nm
• Red: 620 - 780 nm
• When white light passes through or is reflected
by a colored substance, a characteristic portion
of the mixed wavelengths is absorbed. The
remaining light will then assume the
complementary color to the wavelength(s)
absorbed.
 Light and the observation of colors

Suppose we shine a beam of white light

at a substance that absorbs 420-430 nm
region (violet color), the light that is
transmitted is mostly yellow, the
complementary color. This yellow light
reaches our eyes, and we “see” the
substance as a yellow colored
substance.
 The amount of light absorbed is related to the concentration of the analyte.
Conc of analyte  Aborbance
Example 1: KMnO4 (Mn = +7)
The color in KMnO4 arises from an electronic transition, but it is
actually not a d-d transition, since the Mn in this compound has no
d electrons. It arises from a charge transfer (LMCT) within the
molecule, in which photons promote an electron from the highest
energy molecular orbital in one of the Mn-O bonds to an empty d
orbital on the manganese. This promotion is equivalent to the energy
of a yellow photon, so yellow light is absorbed leaving us to see
purple or violet, the complementary color.

Example 2: Cu2+ solution

Appears blue, because it absorbs the complementary color
orange from white light and transmits the remaining blue light.
The more concentrated Cu2+, the more orange light is absorbed
and the deeper the resulting blue color of the solution.
Give Reason:

• The color of Ru(bpy)32+ Tris(bipyridine)ruthenium(II) is orange

Color absorbed: blue

• The color of Cr3+-EDTA complex is violet

Color absorbed: yellow
Light:
▪ Light has wave and particle properties
▪ It Is electromagnetic waves; consists of two components: Electric and Magnetic
and they are perpendicular to each other
▪ It consists of small units called photons

Wavelength λ, Frequency ν, and Energy, E, of photons:

E = hν h : Planck’s constant 6.626 x 10-34 J.s
E= hc/λ c : speed of light 3 x 1010 cm/sec or 3 x 108 m/s
E= hc ῡ ῡ : wavenumber (cm-1 or m-1)
ῡ = 1/ λ
- Energyis inversely proportional to wavelength but proportional to
frequency and wavenumber
- As the wavelength increases, the energy of the radiation
decreases.
Example 1.1:

The wavelength of the green light photon from a traffic signal is

centered at 525 nm. Calculate the energy in joules ?
(h=6.626 X 10-34 J.s, c = 3 X 108 m/s)?

Solution:
c = 3 X 108 m/s
λ = 525 X 10-9 m = 5.25 X 10-7 m
υ = c / λ = (3 X 108 m/s) / (5.25 X 10-7 m) = 5.71 X 1014 (Hz)
E = hυ = 6.626 X 10-34 X 5.71 X 1014 J/photon
E = 3.78 X 10-19 J/photon
Example 1.2:

By how much kilojoules per mole is the energy of CO2 increased when it
absorbs IR-radiation with ύ = 2300 cm-1?
(NA = 6.022 X 1023 molecules per mole)
Solution:

E = hcύ = 6.626 X 10-34 J.s X 3 x 108 m/s X 2300 cm-1 X 102 m-1
E = 4.572 X 10-20 J/molecule
E = 4.572 x 10-20 X 10-3 kJ X 6.022 X 1023 /mole
E = 27.53 kJ/mole

IR absorption increases the amplitude of vibration of CO2 bonds e.g.

O => C => O
Problems on Chapter 1:

[1] One of the electron transitions in a hydrogen atom

produces infrared light with a wavelength of 7.464 × 10-6 m.
What amount of energy causes this transition?
(h=6.626 X 10-34 J.s, c = 3 X 108 m/s)?

[2] Find the energy in joule per photon of microwave radiation

with a frequency of 7.91 × 1010 Hz.

[3] What is the wavelength in nm of UV light that has an

energy of 2.39 × 10-18 J?

[4] By how much kilojoules per mole is the energy of O2

increased when it absorbs UV- radiation with a λ = 147 nm