Topic;

Principle of atomic fluorescence

spectroscopy

presented by ;
Mujeeba Zahid

Roll No;
41048
What is fluorescence spectroscopy

 Fluorescence spectroscopy is a type of electromagnetic spectroscopy that

analyzes fluorescence from a sample.
 Fluorescence is a photoluminescence process in which atoms or molecules
are excited by absorption of electromagnetic radiation.
 The excited species then relax to the ground state ,giving up their excess
energy as photons..
 Single molecules of selected species have been detected by fluorescence
spectroscopy.
Stages of atomic fluorescence
spectroscopy
 AFS is a two stage process of excitation and emission
 Stage 1: A high intensity monochromatic discharge lamp provides the excitation
energy which is focused onto the analyte atoms.
 Stage 2: The electrons surrounding the atom absorb the energy and are excited to
a higher energy level. This is an unstable state and the electron quickly drops back
to it's original ground state. In doing so, the election loses the energy in the form of
emitted light (fluorescence).
 The short –lived emission that occur is called Fluorescence.
 Luminescence that is much longer lasting is called Phosphorescence.
 Fluorescence Spectroscopy
Singlet
Excited
State (S1) Fluorescence
h
Spin allowed
n
Fast (ns)
Emissio
n
Organic
molecules

Triplet
Excited
State (T1) Phosphorescence
h Spin “forbidden”
n
slow (ms to s)
Emissio
n Transition metal
complexes
What is the basic principle of
fluorescence spectroscopy
 The principle of fluorescence spectroscopy Is based upon Jablonki
diagram.

• The Jablonski diagram is widely used in

fluorescence spectroscopy to illustrate
the excited states of a molecule and the
radiative and non-radiative transitions
that can occur between them.
 Jablonski Diagram
7

S
Excitation
S2
1 T2
Internal Conversion
Fluorescence
Energ T1
y Non-radiative decay
Intersystem Crossing
Phosphorescence
S
0
Energy level diagram shows some of the processes that occur during
fluorescence:”
Relaxation process

 Radiative & Non-Radiative Transitions

 The coloured arrows represent the various transitions that can transfer energy
between the molecular states and are split into radiative and non-radiative
transitions.
 Radiative transitions are transitions between two molecular states where the
energy difference is emitted or absorbed by photons and are represented in a
Jablonski diagram by straight arrows.
 Also called fluorescence emission.
 Non-radiative transitions are transitions between two molecular states without the
absorption or emission of photons and are represented in a Jablonski diagram by
undulating arrows.

The two most important non-radiative
relaxation methods
 Vibrational relaxation
 Depicted by the short wavy arrows between vibrational energy levels, take
place during collisions between excited molecules and molecules of the
solvent.
 Internal Conversion
 Nonradiative relaxation between excited the lower vibrational levels of an
excited electronic state and the higher vibrational levels of another
electronic state can also occur.
 Stokes shift

 Molecular fluorescence bands are mostly made up of lines that are longer
in wavelength
 ,higher in frequency
 , and thus lower in energy
 than the band of absorbed radiation responsible for their excitation.
 This shift to longer wavelength is called stokes shift.
 Stokes –shifted fluorescence is longer in wavelength than the radiation that
caused the excitation.
 Mirror Image Rule

n4
n
S n32
n1
1

n4
n
S n32
n1
0
 Fluorometer

3
1 2
Components
1) Light source
2) Monochrometer 2 4
3) Sample
4) Detector
5) Filters
6) Slits
7) Polarizers
References;

 Skoog and West’s Fundamentals of Analytical chemistry ;(page 761 )

 9th edition
 https://www.edinst.com/blog/jablonski-diagram/
 https://www.azom.com/article.aspx?ArticleID=16086