Introduction to Spectroscopy

Ricardo O. Louro
Oeiras, Outubro 2015
 Spectroscopy
Spectroscopy is the study of matter using electromagnetic radiation

The name derives from the latin word ‘spectron’ which means ghost.

The reason is that spectroscopy does not provide a direct observation of

matter but instead provides a measurement of the effect that matter
imprints in electromagnetic radiation- its ghost.

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 Spectroscopy

Identification of the presence of water in the atmosphere of a planet 120

light-years away.

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 What is electromagnetic radiation?
Electromagnetic radiation has properties of matter and of waves, and
therefore is considered to display wave-particle duality

It can be described as:

An oscillating electric and magnetic field that propagates through

space (waves). The distance between successive peaks of the waves is
called the wavelength.

space

The energy of the radiation is related to the wavelength:

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 What is electromagnetic radiation?

The intensity of the radiation is given by the maximum amplitude of the

electric and magnetic waves.

space

ε0 Is the medium permitivity and µ0 is the permeability, E0 and B0 are the

electric and magnetic field intensities.

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 What is electromagnetic radiation?
Electromagnetic radiation has properties of matter and of waves, and
therefore is considered to display wave-particle duality

It can be described as:

An oscillating electric and magnetic field that propagates with time

(waves). The number of waves per unit time is called frequency.

time

The energy of the radiation is related to the frequency:

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 What is electromagnetic radiation?
Electromagnetic radiation has properties of matter and of waves, and
therefore is considered to display wave-particle duality

It can be described as:

Discrete packets called photons that move through vacuum at the

speed of 2.998x108m/s. The intensity is given by the number of photons
per unit of time.

Exercise
How many photons per second does the red pointer emit, considering that
it emits 1 mW at 650 nm?

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Why does electromagnetic radiation interact with
matter?

It is known since the beginning of the

20th century that matter and energy are
two manifestations of the same aspect
of reality.

E = m.c2

Albert Eistein
1879-1955

When spectroscopy is performed, the component on the left side of the

equation is used to obtain information on the component on the right
side, by measuring some aspect of their interaction.

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What happens when electromagnetic radiation and
matter interact?
It can be absorbed or reflected- the basis for color when we are dealing
with electromagnetic radiation in the visible range.

750nm 400nm

Matter can emit electromagnetic radiation. This emission can occur

spontaneously as in the case of radioactive decay of atomic nuclei. The
emission can be stimulated giving rise to:

-fluorescence when emission arises from an excited singlet state

-phosphorescence when emission arises from an excited triplet state.

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What happens when electromagnetic radiation and
matter interact?

The plane of oscillation of the electromagnetic waves can be rotated, by

molecules that are inherently chiral.
Light that is left hand polarized and light
that is right hand polarized cross a
material at different speeds giving rise
to the phenomena of optical activity and
circular dichroism. This is used to
identify optical isomers and secondary
structures of proteins.

Matter can slow down the propagation

of electromagnetic radiation leading to
the phenomenon of refraction.

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What happens when electromagnetic radiation and
matter interact?

Matter can promote the diffraction of

electromagnetic radiation. This occurs
because electromagnetic radiation
behaves as waves that can interact giving
rise to constructive and destructive
interference patterns. These patterns can
be interpreted to build models of the shape
of the obstacles that were in the path of the
radiation. Diffraction is the basis for the
determination of the 3D structure of
molecules up to atomic resolution using X-
ray radiation.

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What happens when electromagnetic radiation and
matter interact?

Electromagnetic radiation can be scattered. This

scattering can be elastic (Rayleigh) or inelastic
(Raman). From the pattern of scattering the size
and shape of biological macromolecules can be
reconstructed. This is the basis of the methods
such as Dynamic Light Scattering (DLS) or Small
Angle X-ray Scattering (SAXS).

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What information can be obtained from matter using
electromagnetic radiation?

The information that can be obtained depends on the nature of the

sample and on the energy of the electromagnetic radiation.

The energy of electromagnetic radiation is given (in Joule) by:

E = h.ν
h is Plank’s Constant (6.62608x10-34 Js)
ν is the frequency of oscillation of the electric and magnetic fields (Hz)

It can also be expressed as:

E = h.c/λ or E = h.c.ν
Where λ is the vacuum wavelength (in meter) and ν is the vacuum
wavenumber (typically expressed in cm-1)

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 The electromagnetic spectrum
Historically the EM spectrum was divided into regions that span specific
frequency ranges.

Human activity uses the electromagnetic spectrum from the highest

energy range (gamma rays in PET scans) to the lowest energy range
(radio waves for communication).

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 The electromagnetic spectrum
Different aspects of the matter are affected depending on the frequency
of the radiation

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What is the nature of the interaction between matter
and EM radiation?

The view according to classical physics:

-light is an electrical and magnetic field that oscillate in time and space

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What is the nature of the interaction between matter
and EM radiation?
The view according to classical physics:

-matter behaves as an harmonic oscillator: when displaced from

equilibrium it experiences a restoring Force that is proportion to the
magnitude of the Displacement.

F= - k x
It has a spring constant k

It has a damping constant γ

It has a cycle frequency ω

It has a mass and charge m, q

The oscillating electrical and magnetic force fields drive the harmonic
oscillator 17
Does it make sense to treat matter as an harmonic
oscillator?

Electrons when taken from their ground state in orbitals tend to return.

Chemical bonds when stretched or bent tend to return to their

equilibrium shape and length.

Magnetic nuclei align their spins (magnetic moment) with an external

magnetic field and if disturbed tend to return to this favored orientation.

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 What are the limitations?

Just like a real spring can be stretched beyond a length where it will no
longer recover, when dealing with the interaction between EM radiation
and matter the same occurs.

E(q)
Just like a real spring cannot
be compressed beyond a
point, when dealing with the
interaction between EM
radiation and matter the E(q)
same occurs.

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 How does the interaction occur?

For matter treated like an harmonic oscillator, the interaction with EM

radiation has the same effect as applying an external force.

The interaction is more efficient when the frequency of oscillation of the

matter is the same as the frequency of oscillation of the EM radiation
(the condition of Resonance)

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 Resonance

In the condition of resonance there is a transfer of power from the EM

radiation to the matter leading to greater displacement (absorption of
energy). The resonance frequency is related with the mass and the
spring constant:

m1.m2
µ=
m1 + m2

The reduced mass takes into account the unequal distribution of mass
over a bond.

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 Exercise

Given that the resonance frequency ω0 is related with the mass of the
harmonic oscillator what do you expect to happen when you replace
hydrogens for deuteriums in a spectroscopically active molecule?

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What happens to the matter after the application of the
electromagnetic radiation?

Just like a real spring after being disturbed will display a damped
oscillation until settling back to the equilibrium the same happens with
the interaction between matter and EM radiation.

time

A is the wave amplitude, and γ is the damping rate constant.

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 What is the nature of damping?

A process that is time dependent and influences the amplitude or phase

of oscillation and occurs with loss of energy of the system to the
environment.

This can occur by:

Interaction with the environment (friction is the classical mechanical

analogue)

Emission of radiation

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 How is the power transferred?

A damped harmonic oscillator absorbs power according to a

mathematical shape that is called a Lorenzian line

ω0 is the resonance frequency and γ is the constant of the exponential

decay of oscillation.

At mid height of the power absorption peak, the width (in units of
frequency) is equal to twice the exponential decay constant.
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 What we know

- how power is transferred and

- how it is dissipated

This information defines the position and shape of the spectroscopic

lines

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 How much power is transferred?

A UV-visible absorption spectrum of a real protein

0.4

0.35

0.3

0.25
Abs.

0.2

0.15

0.1

0.05

0
250 300 350 400 450 500 550 600
λ / nm

The spectrum shows that different lines display different intensity.

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 How much power is transferred?

If the interaction of EM radiation with mater is like applying an external

force to a swing, what is swinging?

- In the case of the electric field component of EM radiation, charges will

be moved. A neutral body made of charged particles can be affected as
long as it possesses a dipole moment
W = - µ.E

Where E is the electrical field strength of the radiation and µ is the dipole
moment operator.
µ=Σqr
Because the EM radiation does not create or destroy charges, it is clear
that the intensity of power transfer is larger for larger charge
displacements .

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 How much power is transferred?

When the bonds between charges are aligned with the direction of
propagation of the field, the oscillating field will stretch or compress the
bonds.

If the alignment is not perfect, the power transfer is less efficient. This
reduction of efficiency is given by the cosine of the angle between the
dipole moment and the direction of propagation of the field.

In analogy with the swing, the parent stands aligned with the swing.

Since the resonance frequency and intensity of power transfer depend

on the nature of the nuclei and the electronic distribution, a quantitative
treatment requires quantum mechanics

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30
 What is the origin of quantum mechanics?

The glow emitted by heated objects can not be explained by classical

physics.

8πν 2 The Rayleigh-Jeans law only matches

kT
c3 experimental observation in the infrared range.

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 What is the origin of quantum mechanics?

Max Panck developed an explanation that implicates that the energy of

the harmonic oscillators assumes discrete values and is proportional to
an integer multiple of the oscillation frequency

E=nhν
This allows the determination
of the radiation density, ρ(νji) for any
chosen frequency that
matches the black body
radiation emission spectrum.

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 What is the origin of quantum mechanics?

The photoelectric effect can not be explained by classical physics.

When a metallic surface is illuminated electrons can be ejected

-light below a certain frequency cannot eject electrons no matter how

intense

-light above the threshold frequency will always eject electrons no mater
how weak

-the kinetic energy of the electrons increases linearly with frequency

above the threshold frequency. The slope equals the Planck constant
and does not depend on the intensity of light

E = h.ν

Albert Einstein won the 1921 physics Nobel Prize by explaining the
effect based on a corpuscular theory of light –the photon was born
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 What are the differences between the Quantum
Mechanical and Classical description?
In the classical description particles travel along a defined trajectory

In the quantum mechanical description, derived from the de Broglie relation,

particles are characterized by a wavefunction Ψ instead of a trajectory.
h
λ=
m.v

Louis de Broglie
(1892-1987)

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 What are the differences between the Quantum
Mechanical and Classical description?

In the classical description there are no restrictions to the energy that can be
exchanged in a interaction between matter and EM radiation

In the quantum mechanical description the energy that can be exchanged in

an interaction between matter and EM radiation has discrete specific values

These values correspond to the transitions between specific states of the

matter that interacts with the EM radiation. These are called the intrinsic- or
eigenstates of the matter.

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 What are the properties of eigenstates?
The quantum eigenstates, being characteristic of the matter, are time
independent.

These states correspond to specific characteristics of the matter: vibration,

rotation, translational, electronic, nuclear, spin. In order to know which
eigenstate we are referring to, we assign a number to each discrete condition
that can be assumed by these characteristics of the matter: these are called
the quantum numbers

Eigenstates described by different quantum numbers have different energy

levels. The energy is determined by the Schrödinger equation in the time
independent version

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 What is the Schrödinger equation?

The Schrödinger equation provides

a way to determine the energy of a
particle with a given wavefunction.

It is generally written in the compact form

EΨ = HΨ
Where Ψ is the wavefunction to
be determined, E is the energy,
and H is an operator called the Erwin Schrödinger
Hamiltonian. 1887-1961

Recall the equation for the harmonic oscillator

E(q) = ½ k Q 2
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 In which eigenstates do we find the matter?

Since the eigenstates have different energy, the Boltzmann distribution allows
us to know the population fraction of each state that we find at a given
temperature.
Nj  − (E j − Ei ) 
= exp 
Ni  kT 
Ej
Ej

Ei
Ei

EM radiation couples the eigenstates. If there is a spectroscopic transition the

fraction of the various eigenstates changes and energy is absorbed or
released by matter.

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 In which eigenstates do we find the matter?

Because the population fraction of the various eigenstates is being modified,

investigation of this process requires the application of the time-dependent
Schrödinger equation

The larger the population difference between the eigenstates before the
interaction with the EM the larger the maximum energy that can be
transferred.
Ej
Ej

Ei
Ei

When the populations become equal no further transfer occurs

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 How are the population fractions modified?
The equilibrium distribution of the eigenstates can be modified by three
phenomena as proposed by Albert Einstein:

-stimulated absorption of radiation

-stimulated emission of radiation

-spontaneous emission of radiation

The frequency of the radiation that

can be absorbed or emitted is defined
by the energy separation between
the eigenstates

A partial representation of the Jablonski diagram 40

 Stimulated absorption

ρ(νji) is a measure of how much of the

radiation has a frequency corresponding to
the energy difference between the i and j
states: the radiation density at the correct
frequency

Bji is the rate constant for the process and

is called the Einstein coefficient for
stimulated absorption

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 Stimulated emission

Bij is the rate constant for the process and

in this case is called the Einstein
coefficient for stimulated emission

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 Spontaneous emission

Aji is the rate constant for the process and

in this case is called the Einstein
coefficient for spontaneous emission,
because it is independent of the amount of
radiation present.

All three processes are first order, which

means that they have an exponential
dependence with time.
What is the consequence?
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 Equilibrium is reached

Equilibrium is reached when the rate of absorption equals the sum of

rates of emission

This allows the A and B Einstein coefficients to be related.

Rearranging the expression and knowing that the radiation density ρ(νji)
can be obtained from Planck’s expression (slide 28):

This expression is written in a way that shows that spontaneous emission

becomes more important than induced emission with increasing
frequency, but we still do not know what are the values of A and B.

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 The transition moment
Moment characterizes a particle that is in motion

When matter interacts with EM radiation the motion is changed

In quantum mechanics wavefunctions replaced the trajectories of

classical mechanics

Similarly to what was made in the classical case, considering the

electrical component of the radiation

The transition moment µji defines the changes between the

wavefunctions of the starting and finishing states
µji

The transition probability is the | µji |2 and relates to the Einstein

coefficients by:
|µji |2
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 What are the consequences?
The solution of the expression of the transition moment defines the
conditions in which B and A are different from zero. If the Einstein
coefficients are different from zero these transitions can take place.

This is the origin of the quantum mechanical selection rules, that

define which transitions can actually be observed in any kind of
spectroscopy.

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How does all of this relate with the question that
brought us here?
The maximum amount of energy transferred is related with the initial
population difference between the states

The rate of the interactions between matter and EM radiation is given by

the thermal population of the eigenstates and the value of the Einstein
coefficients

The value of the Einstein coefficients determine what is the population

ratio at equilibrium

The Einstein coefficients are related to the transition moment which is the
source of the selection rules for the quantum mechanical transitions.

The selection rules state which of all possible transitions between

eigenstates can actually happen

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 How does this relate with ‘real’ life?

When dealing with many molecules that interact with EM radiation, the
radiation that is absorbed by one molecule will not reach the others. The
molecules are said to display an absorption cross section which is a
function of the transition probability for each .

σ = f( |µji|2)
Because of the absorption cross section the intensity of the EM radiation
that emerges from a sample is lower than the irradiation intensity
according to:

Where I is the radiation intensity, l is the sample thickness and N is the

density of molecules

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 The Lambert-Beer law

This equation can be written in a more familiar way:

Which is equivalent to the expression of the Lambert-Beer law

Which is a special case for dilute solutions

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 Limitations of the Lambert-Beer law

Deviations to a linear relationship between absorption and concentration

can arise from three different origins:

Fundamental

Chemical

Instrumental

Fundamental: the law is only valid for dilute solutions for two reasons:

-at high concentration the solute particles do not behave independently.

-at high concentration the refractive index of the sample may vary.

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 Limitations of the Lambert-Beer law

Chemical: The solute must not be engaged in a chemical reaction

otherwise the absorbance can change with time or concentration.

Instrumental: there are two aspects of the instrument that cause deviation
from linearity:

-the radiation is not strictly monochromatic. Polychromatic radiation always

gives negative deviation to the Lambert-Beer law

-intrusion of stray radiation. As for the case of polychromatic radiation this

gives origin to negative deviations.

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 Conclusion

We have determined, from quantum mechanical considerations:

-The frequency of interaction of EM radiation with matter

- The shape of the absorption or emission bands
- The intensity of those bands

-We have reached a general expression that is the basis for the special
case of dilute solutions known as the Lambert-Beer Law.

-We have looked into sources of deviation from the Lambert-Beer law

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 For what can we use spectroscopy?
Spectroscopic Techniques Transition Approximate Typical Wavelength
Region energy (kcal/mol) (cm)
γ-ray Mössbauer Nuclear Spin State 3 x 108 10-11
X-ray X-ray diffraction and 3 x 105 10-8
scattering
Far UV Electronic spectra Electronic 3 x 103 10-5
Near UV Electronic Spectra Electronic 102 3 x 10-5
Carbon Carbon bond energy
Visible Electronic Spectra Electronic 5 x 10 6 x 10-5
IR Vibrational spectra Vibrational 3 x 100 10-3
RT at room temperature
Far IR Vibrational spectra Vibrational 3 x 10-1 10-2
Microwave Rotational Spectra Rotational 3 x 10-2 10-1
Microwave Electron Paramagnetic Electronic Spin 3 x 10-3 100
Resonance
Radio Frequency Nuclear Magnetic Nuclear spin 3 x 10-4 10
Resonance

γ-ray X-ray UV Visible IR Microwave Radiofrequency

Energy l λ
Vibrational Rotational
Nuclear Electronic EPR NMR
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Nuclear Energies Chemical Energies Molecular Energies Spin Energies
54
 Bibliography
This presentation was prepared adapting materials and taking
assorted images from the following references:

P.W. Atkins, Physical Chemistry, 4th Ed, Oxford University Press

D.A. McQuarrie, J.D. Simon, Physical Chemistry a molecular

approach, University Science Books

L. Fusina, Lecture notes, Introduction to spectroscopy, Universitá

di Bologna

D. Nocera, Lecture Notes, Inorganic Chemistry, MIT

G. Elert ,The physics hypertextbook

D. Harvey, Analytical chemistry 2.0; online at asdlib.org

R.R. Crichton & R. O. Louro, Practical approaches to biological

inorganic chemistry, Elsevier
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Further reading

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