Introduction to basic concepts

of fluorescence and
phosphorescence
Class 5
Optics and spectroscopy
Prof. Carlos Augusto Galíndez
Theory of fluorescence and phosphorescence
Fluorescence can be found in the
three basic states of matter:

● Solid
● Liquid
time~10-8 s (10 ns)
● Gas, e.g. Atomic spectroscopy

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Theory of fluorescence and phosphorescence

Fluorescence emitted radiation:

● Without change of frequency, it’s

called resonance radiation or
resonance fluorescence
● Change of frequency toward
larger wavelength (stokes shift)

λ1>λ0

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Excited states producing fluorescence and
phosphorescence
Electron spin:
Paired spins: Molecules do not
exhibit net magnetic field, these
species are called diamagnetic.

Unpaired spins: Free radical have

unpaired electrons and have a net
magnetic moment, this species are
also called paramagnetic.

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Excited states producing fluorescence and
phosphorescence

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Excited states producing fluorescence and
phosphorescence
Singlet and triplet excited states
A molecular electronic state in which
all electrons spin are paired is called
singlet state (no splitting of
electronic stated under magnetic
field)

Ground state for free radical is called

doublet state because there are two
possible orientations for the spin (up
or down)
They have different energies!
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Excited states producing fluorescence and
phosphorescence
Singlet and triplet excited
states Excited singlet state:
Spins remains paired

When one of a pair of electrons of The molecule is still

diamagnetic!
a molecule is excited to a higher
energy level. a singlet or a triplet
state is formed.

singlet state, the spin of the

promoted electron is still paired
with the ground-state electron Ground singlet state

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Excited states producing fluorescence and
phosphorescence

Excited triplet state:

Singlet and triplet excited Spins become unpaired
states
The molecule becomes
paramagnetic!
In the triplet state, however, the
spins of the two electrons have
become unpaired and are thus
parallel.

Ground singlet state

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Excited states producing fluorescence and
phosphorescence

ACN Molecule

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Excited states producing fluorescence and
phosphorescence Jablonski diagram
Singlet and triplet excited
states
Singlet to triplet transition is less
probable than singlet to singlet
transition.

Lifetime in excited triplet state:

~10-4 s
Absorption band
Lifetime in excited singlet state: is less intense
than fluorescence
~10-8 s band

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Excited states producing fluorescence and
phosphorescence

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 Deactivation processes
Radiation process
Involves the emission of a photon
➔ Fluorescence
➔ Vibrational relaxation (Is fast ~ ps) is
➔ Phosphorescence
Rule: The favored route to the ground
state is the one that minimizes the
lifetime of the excited state
Radiationless process
Does not involve the emission of a photon
mediated by collisions between excited
species and solvent molecules.
Consequence: Fluorescence band is
shifted toward longer wavelengths
This process is evidenced by
raising the temperature.
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 Deactivation processes
Radiationless
process
Quinone
Does not involve the emission of a photon
S0→S2
➔ Internal conversion a molecule passes
to a lower electronic energy state
without emission of radiation. S0→S1
S1→S0
Involves states of the same multiplicity
(e.g. singlet to singlet or triplet to triplet)

It is efficient when there are overlapping

vibrational levels of different electronic
levels.

Both excitation band at 250 or 350 nm provides the same

emission band centered at 450 nm due to internal conversion
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 Deactivation processes
Radiationless
process

Does not involve the emission of a photon

➔ External conversion involve interaction ➔ Intersystem crossing is a process in

and energy transfer between the excited which there is a crossover between
molecule and the solvent or other electronic states of different multiplicity.
solutes. S1→ T1

Experimental evidence: Changing the The probability of intersystem crossing

solvent may affect the intensity of is enhanced if the vibrational levels of
fluorescence band the two states overlaps.

Low temperature and high viscosity tend The heavy-atom effect also increases the
to enhanced fluorescence. probability of intersystem crossing (e.g.
Iodine, Bromine, etc.)
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 Variables affecting fluorescence and
phosphorescence
Quantum efficiency of fluorescence

Is simply the ratio of the number of molecules

that luminesce to the total number of excited Molecular structure & Environment factors:
molecules. affect the intensity of emission.

● kpd and kd depends on molecular

structure (important for radiation with
λ<250 nm)
fluorescence ● ki, kec, kic are highly influenced by the
environment.

dissociation

Intersystem External Internal Predissociation

crossing conversion conversion

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 Variables affecting fluorescence and
phosphorescence

Transition types in fluorescence

σ*→σ (too energetic, is seldom observed to

cause fluorescence)

Fluorescence emission is frequently observed

for less energetic transitions:
π*→π (Higher quantum efficiency)
π*→n

Molar absorptivity: (π*→π ) > (π*→n)

Lifetime: 10-7~10-9 s vs 10-5~10-7 s

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 Variables affecting fluorescence and
phosphorescence

Transition types in fluorescence

σ*→σ (too energetic, is seldom observed to

cause fluorescence)

Fluorescence emission is frequently observed

for less energetic transitions:
π*→π (Higher quantum efficiency)
π*→n

Molar absorptivity: (π*→π ) > (π*→n)

Lifetime: 10-7~10-9 s vs 10-5~10-7 s

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 Variables affecting fluorescence and
phosphorescence
Fluorescence and structure

The most intense and the most useful

fluorescence is found in compounds
containing aromatic functional groups with
low-energy π*→π transitions.

● Compounds containing aliphatic or

alicyclic carbonyl groups, e.g. aliphatic
ketones.
● Compounds containing conjugated
double bond

The number of these system exhibiting

fluorescence is less than the number of
aromatic systems. O'Sullivan, M. P., & Testa, A. C. (1970). Fluorescence of aliphatic
ketones. Journal of the American Chemical Society, 92(20), 5842-
5844.

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 Variables affecting fluorescence and
phosphorescence
Fluorescence and structure

Quinoline

Pyridine Furan Isoquinoline

Indole
Chlorophyll A
Thiophene Pyrrol
the quantum efficiency usually increasing with the
Does not exhibit fluorescence! number of rings.
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 Variables affecting fluorescence and
phosphorescence
Fluorescence and structure

Substitution of a carboxylic acid or carbonyl group on an aromatic ring generally inhibits fluorescence.
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 Variables affecting fluorescence and
phosphorescence
Effect of structural rigidity

It is found empirically that fluorescence is particularly favored in molecules with rigid structures.
Lack of rigidity in a molecule probably causes an enhanced internal conversion rate.

Provides rigidity, the

molecule lie in a plane

Fluorene Biphenyl

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 Variables affecting fluorescence and
phosphorescence
Temperature and solvent effect

Quantum efficiency decreases with

temperature increase because the frequency of
collision increases and external conversion can
occur.
Decrease of solvent viscosity increase the
likelihood of external conversion.
How to promove phosphorescence?
Compounds containing heavy atoms are
frequently incorporated into solvents when
enhanced phosphorescence is desired.

solvents containing heavy atoms or other

solutes with such atoms in their structure also
decreases fluorescence.

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 Variables affecting fluorescence and
phosphorescence
Effect of pH on fluorescence

More stable first excited state

(more resonance structures)

The fluorescence of an aromatic compound

with acidic or basic ring substituents is usually
pH dependent.

wavelength and the emission intensity are

likely to be different for the protonated and
unprotonated forms of the compound.

Exhibit
No fluorescence
fluorescence

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 Variables affecting fluorescence and
phosphorescence
Effect of concentration on fluorescence

The power of fluorescence emission F is To relate F to the concentration c of the

proportional to the radiant power of the excitation fluorescing species, we write Beer's law
beam that is absorbed by the system
Power of the beam after its
pass through a medium of
length b

Quantum efficiency Power of the

of fluorescence is incident beam
constant
Constant dependent on
geometry and other factors
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Variables affecting fluorescence and
phosphorescence
Effect of concentration on fluorescence

A plot of the fluorescence radiant power

versus concentration should be linear at
Neglecting higher order terms low concentrations.

When c becomes great enough that the

absorbance is larger than about 0.05, the
higher-order terms in equation become
important and linearity is lost

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Sketch of an instrument for fluorescence
measurement

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Applications fluorescence and phosphorescence
Chemiluminescence

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Applications fluorescence and phosphorescence

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