What is Spectroscopy?

To study changes in the property of light

when it interacts with the matter.
How do we study changes in the property of light when it interacts
with the matter?

MATTER*

Emission
Spectroscopy

Detector

MATTER Absorption
Spectroscopy

Detector
Light Source

Atoms
Atomic Spectroscopy
Molecules
Molecular Spectroscopy
Electromagnetic Spectrum
Types of molecular spectra

Electronic or UV-Vis. Spectra

(Electronic spectroscopy)

Vibrational spectra or vibrational-

Rotational spectra or IR spectra
(IR/vibrational spectroscopy)

Rotational Spectra
(Rotational/microwave
spectroscopy)
ESR spectra (ESR spectroscopy)
NMR spectra (NMR
spectroscopy
 S p e c tro s c o p y

A to m ic M o le c u la r
s p e c tro s c o p y s p e c tro s c o p y

A b s o rp tio n E m is s io n
A to m s w h e n a b s o rb e n e rg y
e le c tro n ju m p to h ig h e r
e n e rg y le v e l a n d th e n ju m p
b a c k b y e m is s io n o f e n e rg y
e q u a l to th e e n e rg y d iffe re n t
b e tw e e n tw o e n e rg y le ve ls
A b s o rp tio n E m is s io n
M o le c u le s , w h e n a b s o rb
e n e rg y tra n s itio n s ta k e s
p la c e b e tw e e n m o le c u la r
e n e rg y le ve ls
M o le c u la r s p e c tru m

A to m ic a b s o rp tio n F la m e e m is s io n
U V -vis ib le & IR
Absorption and emission spectra

Excited State

Ground State
Lambert Beer’s Law / Beer’s Law

I0 I

cuvette
x
• When a beam of monochromatic radiation passes through a solution of
an absorbing substance, then, rate of decrease of intensity of radiation
with the thickness of absorbing medium is directly proportional to
intensity of incident radiations and concentration of solution.
(OR)
• The intensity of a beam of monochromatic radiations decreases
exponentially with increase in the thickness and the concentration of the
absorbing solution
 I0 = Ia + It ; Ia = I0 – It
The probability that the photons of a beam of
intensity I will be absorbed by the sample is
directly proportional to the concentration and the
thickness of the absorbing solution.
dl/I = - α c dx
where dI is the change in intensity produced by the absorption of
radiation on passing through a thickness dx of the solution of
concentration c and α is the proportionality constant.
The minus sign is introduced because there is reduction in intensity.

Applying limits I = I0 at X=0 and I = I at X = b to the

above differential gives

Thus, Beer- Lambert law states that intensity (I) of a beam of

monochromatic light decreases exponentially with increase in the
thickness (x) and the concentration c of the absorbing medium.
Beer- Lambert law states that intensity (I) of a beam of monochromatic light
decreases exponentially with increase in the thickness (x) and the concentration
c of the absorbing medium.

Ln (I/I0) = 2.303 Log (I/I0) = - α b c

Ln (I/I0) = Log (I/I0) = - (α / 2.303) b c

Log (I/I0) = - ε b c A=εlc

Log (I0/I) = ε b c (If Log (I0/I) = A)

A=εbc
ε
Absorption Coefficient or Extinction Coefficient of the absorbing
medium. ( Characteristic of the solute and depends upon the
nature of the solvent / medium , temperature and the wave length
of the light used.)
 Log I/Io = -cx
Log Io/I = cx

A=Log I0/I = cx

A=abc

Abs

Conc.
Absorbance:
• Logarithmic ratio of the intensity of incident radiation to
the intensity of transmitted radiation

• I0 – Intensity of incident radiation

• I - intensity of transmitted radiation
Transmittance:
• The ratio of the intensity of transmitted radiation to the
intensity of incident radiation
T = I / I0

Relation between Absorbance and Transmittance:

A = Log 1/T
 A=εlc
Molar Absorption Coefficient or Molar Extinction Coefficient
If concentration c is expressed in mol dm-3 and the path length l in cm
then ε = dm3 mol-l cm-l
Transmittance (T) = I/ I0 [ If A = Log (I0/I) ]
A = - Log T
T = 10-A = 10 – ε l c
 Limitations
Beer’s law is valid in the following conditions

1. Beer’s law is applicable to dilute solutions only

2. Monochromatic radiations

3. Solute must not undergo association, dissociation,

polymerization, hydrolysis in the solvent

MeBI (MeBI)2 (MeBI)3

660nm 610nm 500nm
1x10-5M 1x10-5M-5x 10-3M 5x10-3M – above
 How do we use Beer-Lambert Law ?
1. Numerical
A monochromatic radiation is incident on a solution of 0.05 molar concentration of an absorbing
substance. The intensity of the radiation is reduced to one fourth of the initial value after passing
through 10 cm length of the solution. Calculate the molar extinction coefficient of the
substance………..

2. Finding the unknown A=εlc

Blank I II III IV V

---

Unknown-1 UnKn-2 UnKn # n

 Flame
Photometry
How many of you have done Flame Photometry
ever in life?
 Atomic Emission and
Atomic Absorption
Spectroscopy
• In atomic emission and absorption spectroscopic technique sample solution
is aspirated into a flame that is hot enough to break the ions or molecules
into their atomic states.

• The concentration of the analyte in the flame can be measured by either

through absorption or emission of the radiation.

• The absorption mode is known as atomic absorption spectroscopy (AAS)

whereas emission mode as flame emission Spectroscopy (FES) or Atomic
emission spectroscopy.
Upper Diagram shows FES while the lower one shows AAS. In FES the flame provides
energy for atomization and excitation, but in AAS it provides only atomization .
 Flame emission spectroscopy

• Flame emission spectroscopy (flame photometry) is a special area of emission

spectroscopy in which a flame is used to excite the atoms.

• For a few elements, such as the alkali metals Na and K, flame is hot
enough not only to produce ground state atoms, but to raise some of the atoms
to an excited electronic state.

• So Flame emission spectroscopy is used for the detection of alkali metals and
some of the alkaline earth metals.
 Flame emission spectroscopy
Sequence of events in flame

Nebulization Desolvation Volatilization

Sample

solutio
n Spray Heat Dry aerosol Free
atoms

• Heat
• Atomic absorption and atomic emission techniques involve introduction of
• sample solution into a flame.
• The solution of the metal salt in question is sprayed into the flame.
• Solvent evaporates leaving the finely powdered salt.
• Vaporization of the salt.
• Conversion of ions into free gaseous atoms.
• Valence electron ( ) is raised to a higher energy state.
 Flame emission spectroscopy
Nebulization – Conversion
of sample
solution in fine
droplets – aerosol
formation which is aspirated to flame
Desolvation – Removal of solvent leaving dry sample

Sublimation- Transition of salt from solid to gaseous state Atomization – Conversion of

ions in to atoms

Excitation – Valence electrons of atoms get excited to higher energy state

Relaxation – Excited electrons relaxes from higher energy state to ground state
Measurement – Wavelength and intensity of emitted radiations is measured
 Flame emission spectroscopy

Principle
•Absorption of heat energy by ground state
atom present in the flame
results in the excitation of valence electron of atoms.
•This valence electron comes back to ground state with the emission
of photon.
•Wave length and intensity of emitted photon helps in
qualitative and
quantitative analysis of the sample.
The energy emitted when this electron drops down into a vacant lower level
is given off as radiant energy of a wavelength determined by the Planck-
Einstein relationship:
E2-E1 = E = h = hc/ = hc/ E2-E1
 of the emitted radiation is characteristic of atoms of a particular element
from which it is emitted. It tells us about the elements which are present in
the flame
For example,
Na -------> Na* (energy from flame) Na* -------> Na + h(at 589 nm)
How do we determine the concentration in FES?
Boltzmann Distribution : N*/ N0 = A e - ∆E/k T
N* : Number of atoms in excited state( Intensity)
N0 : Number of atoms in ground state( Concentration ? )
∆E : E1-E0 = Difference between two energy states
k : Boltzmann Contant
T : Temprature of flame
A : Constant for particular atom
Thus, TEMPRATURE plays an important part in
FES. = High Intensity( Caution )
High Temprature = High Excitation

“TEMPRATURE OF THE FLAME” depends on a) Fuel & Oxidant

b) Fuel : Oxidant
Ratio
Intensity of radiant energy emitted when the atoms return to the
ground state is proportional to the concentration and is the basis of
flame emission spectroscopy.
 Fuel Oxidant Flame temperature
(0C)
Propane Air 1900 oC

Propane Oxygen 2800 oC

Hydrogen Air 2100 oC

Hydrogen Oxygen 2800 oC

Acetylene Air 2200 0C

Acetylene Oxygen 3000 0C

 Flame Photometry
Limitations
(i) Does not Give Information about the Molecular Form of the Metal : due
to atomization of all the forms of metal.
(ii) FES is not applicable to all the Metal Atoms : more applicable due to alkali
& alkaline earth metals.
(iii) Sometimes Sample Preparation is a Lengthy Process: because only liquid
samples have to be introduced into the flame.
(iv) Cannot be used to determine inert gasses?

Any Questions?

Flame Photometry
 Inductively Coupled Plasma-
Atomic emissionSpectrometry
(ICP-AES)
Is it safe to drink TAP WATER??

NO !!!
Because of the
presence of
different heavy
metals/trace
elements
Some common Techniques for detection
of Trace elemental analysis

AAS AES

ICP-AES
 Basics of AES
Atomic emission spectroscopy (AES) or optical emission spectroscopy uses
quantitative measurement of the optical signals when atoms relaxes from
excited state to ground state to determine analyte concentration.

Excitation sources:
• Flame
• Arcs/sparks
• Direct Current Plasma (DCP)
• Indirect Coupled Plasma (ICP)
The energy emitted is directly proportional
• Microwave induced Plasma (MCP) to the concentration of the analyte present
in the solution.

Plasma is superheated matter – so hot that the electrons are ripped away
from the atoms forming an ionized gas. Plasma is often called “the fourth state
of matter,” along with solid, liquid and gas. Plasma is used for atomization.
Inductively Coupled Plasma-Atomic Emission Spectroscopy
(ICP-AES)
 ICP-AES utilizes plasma as excitation source. A plasma is an electrically neutral,
highly ionized gas that consists of ions, electrons and atoms.

 Induction plasma, also called inductively coupled plasma, is a type of high

temperature plasma generated by electromagnetic induction, usually coupled
with argon gas.

 The Ar plasma is the result of the flow of Ar ions in a very strong, localized radio
field.

 The magnetic field induces an electric current within the gas which creates the
plasma. The plasma can reach temperatures up to 10,000 Kelvin.

 Hot enough to excite most elements so they emit light.

 Hot enough to prevent the formation of most interferences, break down oxides and
eliminate most molecular spectral interferences.

 Sensitive approach and most widely applied to determine trace elements.

 Advantages of ICP-AES
ICP- AES is a type of spectroscopic techniques which is capable of detecting
metals and several non-metals at concentrations as low as parts per billions
(ppb).
Elemental analysis with

Extremely Low Detection

Wide Elemental
Simple Spectra Limits (ppt/ppm) or (ng/L
Coverage
to mg/L)

Fast Analysis times (all High Sensitivity

elements at once) and Reliability

High Throughput Multi-elemental Analysis

(approximately 10 - 40 elements per minute per sample)
Advantage: Sensitive detection of elemental
concentrations at trace levels
 Referrences
Skoog, D.A., Holler, F.J., and Crouch, S.R., Principles of Instrumental Analysis,
Thomson Learning (2007).

Skoog, D.A., West, D.M., Holler, F.J., and Crouch, S.R., Fundamentals of Analytical
Chemistry, Brooks/Cole (2003) 8th ed.

Boss, C.B.; Fredeen, K. J., Concepts, Instrumentation and Techniques in Inductively

Coupled Plasma Optical Emission Spectrometry, Second Edition, The Perkin-Elmer
Corporation, USA.
ATOMIC ABSORPTION
SPECTROSCOPY
 Atomic Absorption Spectroscopy

• Most powerful technique for the determination of

trace metals in solution
• 70-80 elements can be detected
• Determination can be made in the presence of many
other elements
• No specific sample preparation is required
• Wide application
 Atomic Absorption Spectroscopy
Principle
A sample solution containing known metal atoms, when introduced
into the flame, is irradiated with light of their own specific
wavelength will absorb light proportional to the density of atoms
in the flame.

Flame

Specific Wavelength
(Hollow cathode lamp) Detector

Thus, AAS measures Amount of Light Absorbed.

Atomic Absorption Spectroscopy
Hollow Cathode Lamp (HCL)

?
 Principle
Absorption of energy by ground state atoms
present in the gaseous state in flame

N*/N0 = A e-/ kT

Receive same  which they emit in the excited state

Absorption density of atoms in flame

Amount of light absorbed concentration of
metallic species
 Differences between AAS and FES

FES AAS
Excitation of atoms – Signal is obtained in presence
emission of photon and and absence of element and
return to ground state. decrease in intensity of signal
Emission intensity is obtained.
measured Absorption is measured
Emission intensity No. of Absorption intensity  No. of
atoms in excited state atoms in ground state
Emission intensity depends Absorption intensity does not
on flame temperature depend on flame temperature
Beer’s law is not obeyed Beer’s law is obeyed over a
over wide range of wide range
concentration
 Advantages of AAS

• AAS - specific – Atom of a particular element can absorb radiation

of their own wavelength – No spectral interference
• Much larger No. of atoms contribute in AAS signal so results are
more authentic
• Variation in flame temperature has less effect
• Absorption intensity does not depends on flame temperature
• 70-80 elements can detected
 Disadvantages of AAS

• Different (HCL) lamp for each element is required

• Elements which for stable oxides eg. Al, Ti, W, Mo, do not give
very good results