King Abdul-Aziz University

Faculty of Science
Chemistry Department

Chem.312
Instrumental Methods of Analysis

Dr.Effat Bahaidarah
[email protected]

Prepared and edited:

Dr. Wejdan Alsaggaf
Dr. Fatmah Al Shareef
Dr. Zainab Saigl

1
Introduction
By the end of this part students will be able to:

• Describe the concept of interaction of electromagnetic

radiation (EMR) with matter.

• Calculate wavelength, wave number, frequency, and energy

of EMR.

• Distinguish between atomic and molecular transition.

• Recognize the types of spectra

Chapter 1 : Introduction
Analytical Chemistry

Quantitative Analysis Qualitative Analysis

Classical Methods
(Wet Chemical Analysis) Instrumental Methods

Gravimetric Analysis Volumetric Analysis

Chromatographic methods
Spectrophotometric methods
Ex: HPLC, GC, IEC
ex: UV-VIS, IR,NMR, MS,
Electrochemical methods Thermal methods AAS, AES, Fluoresecence,X-
ex: voltammetry(Polarography), ray Spectroscopy
Conductometry, Electrogravimetry,
Amperometry)

3
Instruments for analysis
• Converts information stored in the physical or chemical characteristics of the
analyte into useful information.
• Require a source of energy to stimulate measurable response from analyte.

Data domains
The measurement process is aided by a wide variety of devices that convert
information from one form to another. In order to investigate how instruments
function , it is important to understand the way in which information is encoded
(represented) , or transformed from one system of information to another, as a
characteristic of electrical signals .
The various modes of encoding information electrically are called data domains.

4
 Advantages of Instrumental Methods

• Impossibility of analysis using volumetric and

Gravimetric analysis.
• High sensitivity ( ng, pg….)
• Simultaneous analysis
• Huge Number of analysis
• Identification analysis
• Structure analysis

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Selecting an analytical method
Defining the problem
• Accuracy
• Amount of sample
• The concentration range of the analyte
• Interference
• The physical and chemical properties of the sample matrix
• Number of samples

Performance characteristics of instruments

Quantitative characteristics: Qualitative characteristics :
• Precision • Speed
• Bias • Ease and convenience
• Sensitivity • Skill required
• Detection limit • Coast and availability
• Dynamic range • Per-sample cost
• Selectivity 6
Chapter 6 : Introduction to spectrometric methods

➢ Spectroscopy is the study of the interaction between the

electromagnetic radiation and the matter.
➢ Spectrophotometry is the measurement of these interactions i.e.
the measurement of the intensity of light absorbed , emitted or
scattered at a selected wavelength.
➢ The method depends on the light absorbing property of either
the analyte or one of it`s derivatives .
➢ Spectrometry or spectrometric methods refer to the
measurement of the intensity of radiation with photoelectric
transducer or other type of electronic device.
Wave Properties of Electromagnetic Radiation (EMR)

It is a form of energy that has both Wave and Particle Properties

that can be characterized by :
λ is the wavelength of the waves
ν is the frequency of the waves
c is the speed of light

+ Wavelength ()
Electric Field

Amplitude (A)

- Time or Distance
Wave Properties of Electromagnetic Radiation (EMR)

Figure. Wave nature of a beam of single-frequency electromagnetic radiation. In (a). a

plane-polarized wave is shown propagating along the x-axis. The electric field oscillates
in a plane perpendicular to the magnetic field, If the radiation were un polarized, a
component of the electric field would be seen in all planes. In (b). only the electric field
oscillations are shown.

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Wave Properties of Electromagnetic Radiation (EMR)
Wave charctarestics

1- Amplitude (A)
It is the maximum vertical displacement of the wave from normal
position.
2- Wave length (λ ) (nm, μm, cm, m)
It is a measure of distance between two identical adjacent peaks
(high points or crests) or troughs (low points) in a wave.
3- Frequency (v)
It is the number of waves or cycles that
pass a fixed point per second.
The unit of frequency is Hertz (Hz), (s-1)
4- Wave number ( ~)
It is the number of waves in a unit length or distance per cm in
units of cm-1
5- Period (P)
It is the time required for one cycle to pass a fixed point in space. 10
 Wave Properties of Electromagnetic Radiation (EMR)
Wave charctarestics

▪ Radiant Power ( P ): The amount of energy reaching a given area

per second. Unit in watts (W)

▪ Intensity ( I ): The radiant power per unit solid angle.

▪ Absorbance spectrum: A graph of a sample’s absorbance of

electromagnetic radiation versus wavelength.

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The Electromagnetic Spectrum

12
 The Electromagnetic Spectrum
• The EMR spectrum is the entire range of EMR waves in order of
increasing frequency and decreasing wavelength.
The Electromagnetic Spectrum
 Quantum Mechanical Properties of Radiation

The energy of one particle of light (photon) is proportional to its

frequency and can be given by :

E = h
where: E = photon energy (Joules) or (electron volts (eV))
v = frequency (sec-1)
h = Planck’s constant (6.626x10-34J.s)
As frequency (v) increases, energy (E) increases
In vacuum, velocity of light : c =  = 3 x 1010 cm/s
which gives,  =c/ , E = h (c/)

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Example 1:
The wavelength of the sodium D line is 589 nm. What is the
frequency for this line?

Solution :
The frequency of the sodium D line is
Example 2:
What is the energy of a photon from the sodium D line at
589 nm?

Solution :
The photon’s energy is

hc (6.626X10−34J.s)(3X10 8m / s
E= = =3.37X10 -19
J
 589 X 10−9 m
Notes:
• Angstrom unit (A◦) : a unit of length equal to 1 x 10-10
meter.
• Nanometer unit ( nm) : a unit of length equal to 1x10-9
meter.
Regions of the UV, visible and IR spectrum

Region Wavelength Range

UV 180 – 380 nm

Visible 380 – 780 nm

Near-IR 0.78 – 2.5 μm

Mid-IR 2.5 – 50 μm
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 Prefixes for units
Prefix Symbols Multiplier
giga- G 109
mega- M 106
kilo- k 103
deci- d 10-1
centi- c 10-2
milli- m 10-3
micro- μ 10-6
nano- n 10-9
pico- p 10-12
femto- f 10-15
atto- a 10-18
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 Practice Exercises
Exercise (1):
The wavelength of the sodium D line is 589 nm. What are the frequency and the
wavenumber for this line? (υ = 5.09x1014 s-1 , ~ = 1.70 x104 cm-1 )
Exercise (2):
Another historically important series of spectral lines is the Balmer series of
emission lines form hydrogen. One of the lines has a wavelength of 656.3 nm.
What are the frequency and the wavenumber~for this line? (υ = 4.57x1014 s-1 ,

= 1.524 x104 cm-1 )

Exercise (3):
What is the energy of a photon from the sodium D line at 589 nm? ( E= 3.37 × 10-19 J )

Exercise (4):
What is the energy of a photon for the Balmer line at a wavelength of 656.3 nm?
( E= 3.03 × 10-19 J )

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 Practice Exercises

Exercise (5):
Two electromagnetic waves are represented below:

Figure 1
(a) Which wave has the higher frequency? (b) If one wave represents
visible light and the other represents infrared radiation, which wave is
which?
Solution
(a)The lower wave has a longer wavelength (greater distance between peaks).
The longer the wavelength, the lower the frequency (v = c/λ). Thus, the lower
wave has the lower frequency, and the upper wave has the higher frequency.

(b) The electromagnetic spectrum (Figure 1) indicates that infrared radiation has
a longer wavelength than visible light. Thus, the lower wave would be the
infrared radiation.
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 Classification of spectroscopic methods

The various techniques can be classified in several ways,

according to:

1-Nature of the interaction of radiation and matter.

2-Type of EM radiation.
3-Type of material ( matter ).
 Interaction of Radiation and matter

Types of spectroscopy can be distinguished by the nature of the interaction

between the EM radiation and the material.

.
A-Absorption: a transition from a lower level to a higher level with transfer of
energy from the radiation field to the atom or molecule.

B- Emission: a transition from a higher level to a lower level, if energy is

transferred to the radiation field, or nonradiative decay if no radiation is emitted.

C- Scattering: redirection of light due to its interaction with matter, and may or
may not occur with transfer of energy.
 Interaction of Radiation and matter
Absorption
 Interaction of Radiation and matter
Emission and
Chemiluminescence
 Interaction of Radiation and matter
Scattering
 Classification of spectroscopic methods
2- Types of radiative energy

Gamma-ray Spectroscopy
X-ray Spectroscopy
UV-visible Spectroscopy
Infrared Spectroscopy
Laser Spectroscopy
 Classification of spectroscopic methods
3- Types of matter

a) Atoms
Atomic spectroscopy was the first application of spectroscopy
developed. Atomic absorption spectroscopy (AAS) and atomic
emission spectroscopy (AES) involve visible and ultraviolet
light. These absorptions and emissions, often referred to as
atomic spectral lines, are due to electronic transitions of outer
shell electrons as they rise and fall from one electron orbit to
another. Atoms also have distinct x-ray spectra that are
attributable to the excitation of inner shell electrons to excited
states.
 Classification of spectroscopic methods
3- Types of matter

b) Molecules
The combination of atoms into molecules leads to the creation of
Molecular spectra can be obtained due to electron spin states
(electron paramagnetic resonance NMR ), molecular rotations,
molecular vibration and electronic transitions ( uv/vis molecular
spectroscopy ). Rotations are collective motions of the atomic
nuclei and typically lead to spectra in the microwave spectral
regions; rotational spectroscopy and microwave spectroscopy are
synonymous. Vibrations are relative motions of the atomic nuclei
and are studied by both infrared and Raman spectroscopy.
Electronic excitations are studied using visible and ultraviolet
absorption and emission spectroscopy.
 Classification of spectroscopic methods
3- Types of matter
c) Nuclei

Nuclei also have distinct energy states that are

widely separated and lead to gamma ray spectra.
Distinct nuclear spin states can have their energy
separated by a magnetic field, and this allows for
NMR spectroscopy
 Interaction of EMR with matters
Electromagnetic Radiation
Energy Frequency Wavenumbe wavelengt
Radiation Interaction with matter (E) () r (ṽ) h ()

The wavelength decrease

Affect and excite

The wavenumber increase

γ -Ray

The Frequency increase

the atomic nuclei
Effect inner
X - Ray

The energy increase

electronic transition
Effect outer
UV& visible electronic transition

Molecular Vibration
IR

Molecular rotation
Microwave (electronic spin
resonance)
Relationship among Energy, Frequency,
Nuclear magnetic
Radio wave wavenumber and wavelength
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 Absorption of Radiation

Ground and Excited State

❖ Under the normal condition electrons exist in the ground state
but when a chemical absorbs light, these electrons goes from a
low energy state (ground state) to a higher energy state (excited
state)

Energy required of photon to

give this transition:
 =  − 

❖ Only photons with energies exactly equal to the energy

difference between the two electron states will be absorbed.
 Absorption of Radiation

• When a photon of energy hν strikes the atom or molecule, absorption may occur
if the difference in energy, ∆E, between the ground state and the excited state is
equal to the photon’s energy.
• An atom or molecule in an excited state may emit a photon and return to the
ground state. The photon’s energy, hν, equals the difference in energy, ∆E,
between the two states.
 Atomic Absorption
• Atomic transitions involve changes in the electron orbitals. These
changes could be in X-Rays region and cause chances involving
the innermost electron shells of the atom. Changes in outer
electron shells could be in the visible light region of the spectrum.
• The atomic spectrum consists of few lines , therefore it is
sometimes termed line spectrum .

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• E0 – lowest energy electronic level or ground state

• E1, E2 – higher-energy electronic levels
 Molecular Absorption
• The energy of a band in a molecular absorption spectrum is the
sum of three different energy components.
E = E electronic + E vibrational + E rotational

• Transitions between electronic-vibrational-rotational states give

rise to spectra that appear to have bands.

vibration

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rotation
 Absorption Induced by a Magnatic Field
➢ Additional quantized energy levels can be observed as a
result of strong magnetic field. Absorption will appear at
longer wavelength.

➢ Nuclear transitions are between energy levels in the nucleus.

Types of nuclear transitions lead to the emission of alpha,
beta, or gamma radiation.

➢ Absorption by nuclei or by electrons in magnetic fiels is

studied by nuclear magnetic resonance (NMR) and electron
spin resonance (ERS) techniques, respectivly.
 Relaxation Processes

Lifetime of excited atom or molecule is brief because there are

several relaxation processes that permit its return to the ground
state.

Nonradiative Relaxation
Involves lose of energy in small steps as kinetic energy by
collision

Fluorescence and Phosphorescence

Analytically important
Fluorescence more rapidly than phosphorescence
(it will discuss in details later)
 Emission of Radiation
➢ When excited particles relax to lower energy levels, they giving
up their excess energy as photons.
➢ Excitation can be done by:
1. Bombardment with electrons (X-ray emission)
2. Electric excitation (e.g. arc , spark, furnace or plasma) or intense
heat source (e.g. flame) (UV, visible, or IR emission).
3. Beam of electromagnetic radiation (fluorescence).
4. Chemical reaction (chem .
 Emission of Radiation
1- Line spectra

➢ It is a spectrum that consists of narrow, brightly colored, parallel lines on

a dark background
➢ Individual atoms, well separated, in a gas phase

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 Emission of Radiation
2- Band spectra

Vibrational levels

➢ It is produced by molecules. They are the groups of lines which are closely spaced
to one another.
➢ Small molecules and radicals 40
 Emission of Radiation
3- Continuum Spectra

• Produced when solid are heated ( thermal energy) to emit light

• a spectrum that contains or appears to contain all wavelengths but not
spectrum lines over a wide portion of its range.
• Black-body radiation is the type of electromagnetic radiation within or
surrounding a body in thermodynamic equilibrium with its environment.

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 Types of Spectra
1- Absorption spectrum
• A plot of the absorbance of the amount of light absorbed by a sample ( atom or
molecule ) as a function of wavelength or frequency.

2- Emission spectrum
• A plot of the relative power (intensity or abundance) of the emitted radiation as
a function of wavelength or frequency.

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Quantitative Aspects of Spectrochemical Measurements

Key terms and relations:

Transmittance
𝑷
𝑻=
𝑷𝟎
𝑷
%𝑻 = × 𝟏𝟎𝟎
𝑷𝟎

Absorbance
𝑷𝟎
𝑨 = −𝒍𝒐𝒈𝟏𝟎 𝑻 = 𝒍𝒐𝒈
𝑷

P0= incident radiation; P= transmission radiation

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Quantitative Aspects of Spectrochemical Measurements

Beer’s Law:
• Absorbance is directly proportional to the path length, b, the
concentration, c, and a proportionality constant called the
absorptivity, a, or in other words;
A= abc

• When concentration is in Molarity and cell length is in cm, then

a is equal to the molar absorptivity, ε, with units L.mol-1.cm-1
A= εbc

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