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?
 Background and motivation
• Water pollution remains a serious
problem in the United States and in
other industrial countries. The photo
here shows land left over from strip
mining in Belmont County, Ohio.
• The various water pools shown are
contaminated with waste chemicals.
The large pool to the right of center
contains sulphuric acid, and the
smaller pools contain manganese and
cadmium.
• Trace metals in contaminated water
samples are often determined by a
multielement technique such as
inductively coupled plasma atomic
emission spectroscopy.
• Single-element techniques such as
3
atomic absorption spectrometry are
 Atomic spectroscopic methods

• Atomic spectroscopic methods are used for the qualitative and quantitative
determination of more than 70 elements.

• Typically, these methods can detect parts-per-million (ppm) to parts-per-

billion (ppb) amounts, and in some cases, even smaller concentrations.

• Atomic spectroscopic methods are also rapid, convenient, and usually of high
selectivity.

4
 Absorption Spectra
• In atomic absorption spectroscopy, an
external source of radiation impinges
on the analyte vapor.
• If the source radiation is of the
appropriate frequency (wavelength),
it can be absorbed by the analyte
atoms and promote them to excited
states.
• Figure shows three of several
absorption lines for sodium vapor.
The source of these spectral lines is
indicated in the partial energy
diagram shown. In this instance,
absorption of radiation of 285, 330,
and 590 nm excites the single outer
electron of sodium from its ground
state 3s energy level to the excited 3p, 5
 Emission Spectra
• Partial energy level diagram for atomic sodium
showing the source of three of the most prominent
emission lines.
• Before the external energy source is applied, the
sodium atoms are usually in their lowest-energy or
ground state. The applied energy then causes the
atoms to be momentarily in a higher-energy or
excited state.
• External energy (plasma, a flame, a low-pressure
discharge, or by a high-powered laser) promotes
the outer electrons from their ground state 3s
orbitals to 3p, 4p, or 5p excited-state orbitals.
• After a few nanoseconds, the excited atoms relax to
the ground state giving up their energy as photons
of visible or ultraviolet radiation. 6
 Absorption and emission spectra
• After a few nanoseconds, the excited atoms relax to their ground
state by transferring their excess energy to other atoms or
molecules in the medium.
• The absorption and emission spectra for sodium are relatively
simple and consist of only a few lines. For elements that have
several outer electrons that can be excited, absorption and
emission spectra may be much more complex.

7
Electromagnetic Spectrum
 Electromagnetic Radiation

• Electromagnetic radiation consists of a discrete

package of energy known as photons.

• A photon consists of an oscillating electric field and an

oscillating magnetic field which is perpendicular to
each other.

9
Representation of EMR

10
Terminology

11
Contd.

12
EMR and Spectroscopy

13
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
 Spectroscopy

Atomic Molecular
spectroscopy spectroscopy

Absorption Emission Absorption Emission

Atoms when absorb energy M o l e c u l e s , wh e n a b s o r b
electron jump to higher energy transitions takes
energy level and then jump place between m olecular
back by emission of energy energy levels
equal to the energy different
between two energy levels Molecular spectrum

Atomic absorption Flame emission

UV-visible & 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 = -ecx
Log Io/I = ecx

A=Log I0/I = ecx

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
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UCB009: Chemistry

35
UCB009: Chemistry

36
 Last lectures
 Ist lecture: What is spectroscopy?, Atomic and
molecular spectroscopy, Types of spectra, Meaning
of transition

 IInd lecture: Lambert-Beers Law, Limitations,

Deviations, Transmittance, Absorbance

 IIIrd lecture: Lambert-Beers Law questions

Today lecture: Calibration curve method, AES, ICP-

AES

37
Quantitative Analysis: Calibration
Curve Method

38
Lead is extracted from a sample of blood and analyzed at 283 nm and gave an
absorbance of 0.340 in an AA spectrometer. Using the data provided, graph a
calibration curve and find the concentration of lead ions in the blood sample.
[Pb+2] (ppm) Absorbance Calculated Pb (II) concentraions (ppm) Absorbance
0.000 0.000 0.323 0.340
0.100 0.116 • The data provided in the
0.200 0.216
0.300 0.310 problem appears in the
0.400 0.425
0.500 0.520 upper left hand corner of
this MS EXCEL
Lead (II) Calibration Curve worksheet.
0.600 • The graph was used to
0.500
calculate the best fit line.
y = 1.0505x
R² = 0.9988 • The equation was then
0.400
used to calculate the
Absorbance

0.300 concentration of Pb (II)

ions with an absorbance
0.200
of 0.340.
0.100
• The result, 0.323 ppm, is
0.000
displayed above the
0.000 0.100 0.200 0.300 0.400 0.500 0.600 graph.
[Pb+2] (ppm)

39
 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

---
41
 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 (AES).
 Flame Test
Ba Ca Na Li

In FES: Flame provided excitation and atomization energy

In AAS: Flame atomizes only, Lamp is used for excitation

45
Upper Diagram shows FES while the lower one shows AAS. In
FES the flame also provides the excitation, but in AAS it provides
46
only for the 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.
 FES (FP)
 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 it is used for the detection of alkali
metals and some of the alkaline earth metals.
Sequence of events in flame:
 Flame absorption and flame emission techniques involve introduction of a
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.

48
Pictorial Presentation of Sequence of Events

49
 Emission of Radiation
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 = DE = hn = hc/l
l= hc/(E2-E1)
l 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 + hn (at 589 nm)

Sodium vapor exhibits sharp absorption peaks at 589.0 nm due to

excitation of 3s electron to 3p states.
50
 Flame emission spectroscopy
Sequence of events in flame

Nebulization Desolvation Volatilization

Sample
solution

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.

 Wavelength 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 = DE = hn = hc/l l= hc/ E2-E1
l 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 + hn (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 Temprature = High Excitation = High Intensity( Caution )

“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

Q. Sodium atom absorbs at 589 nm. Calculate the energy gap
between the ground and excited state. At a temperature of 2000
K, what is the ratio of excited state to ground state population
(N*/No)? Given that A =2, kB = 1.38 × 10-23 J K-1 mol-1

ΔE= hc/λ = (6.634 × 10-34) (3 × 108)/(589 × 10-9) = 3.3 × 10-19 J

N*/N0 = A Exp(-ΔE/kBT)

N*/N0 = 2 Exp (-3.3 × 10-19/(1.38 × 10-23 × 2000))

N*/N0 = 2 Exp(-11.956) = 1.3 × 10-5

% Population in excited state: 1.3 × 10-5 × 100 = 0.0013

Q. What T is required, to get 100% excitation in the above problem?

Ans: 34506 K @ A = 2
21766 K @ A= 3
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58
59
 Population on Thermal Excitation
Q. Sodium atom absorbs at 589 nm. Calculate the energy gap between
the ground and excited state. At a temperature of 2000 K, what is the
ratio of excited state to ground state population (N*/No)? Given that A
=2, kB = 1.38 × 10-23 J K-1 mol-1

ΔE= hc/λ = (6.634 × 10-34) (3 × 108)/(589 × 10-9) = 3.3 × 10-19 J

N*/N0 = A Exp(-ΔE/kBT)

N*/N0 = 2 Exp (-3.3 × 10-19/(1.38 × 10-23 × 2000))

N*/N0 = 2 Exp(-11.956) = 1.3 × 10-5

% Population in excited state: 1.3 × 10-5 × 100 = 0.0013

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63
 Applications
1. Detection of elements from group I and II .
2. In clinical field, Na+ and K+ ions in body fluids, muscles and
heart can be determined by diluting the blood serum and
aspiration into the flame.
3. Analysis of soft drinks, fruit juices and alcoholic beverages can
also be analyzed by using flame photometry.
4. In agriculture, the fertilizer requirement of the soil is analyzed
by flame test analysis of the soil.
5. Natural and Industrial waste water analysis

64
 Qualitative Analysis
The technique is used to detect elements of group I and II of the periodic table

Within the flame, there are many more atoms in the ground state than in the
excited state.

For Zn, for instance, in a 2000 K flame, there are 7.3 x 1015 atoms in the
ground state for every excited atom.

About 10 elements can be determined by conventional flame emission

spectroscopy, since a flame provides sufficient energy for only a few elements to
have a significant number of atoms in an excited state.

65
 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
 Limitation of Flame Emission Photometry
1-The number of excited atoms in flame is very small. It is the alkali
and alkaline earth metals that can be practically determined.

2-It needs perfect control of flame temperature.

3- Interference by other elements is not easy to be eliminated.

4-less reliable than AAS

5-Does not provide any information about molecular state of compound

6-Non-radiative elements (such as C, H, halides) cannot be detected

67
 Inductively Coupled Plasma-
Atomic Emission Spectrometry
(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 (OES)
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)
• Inductively Coupled Plasma
(ICP) The energy emitted is directly proportional
to the concentration of the analyte present
• Microwave induced Plasma (MCP) 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.

ICP- Sequence of Events
 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)

Thus, AAS measures Amount of Light Absorbed.

 Role of the Hollow Cathode Lamp (Radiation source)

 The cathode contains the element that is analysed.

 Light emitted by hallow-cathode lamp has the same
wavelength as the light absorbed by the analyte element.
 Different lamp required for each element (some are multi-
81
element)
 Working of Hollow Cathode Lamp

• a tungsten anode and a

cylindrical cathode
• neon or argon at a pressure of
o Ionize the inert gas at a potential of ~ 300 V 1 to 5 torr
o Generate a current of ~ 5 to 15 mA as ions and • The cathode is constructed
electrons migrate to the electrodes. of the metal whose spectrum
is desired or served to
support a layer of that metal
o The gaseous cations acquire enough kinetic energy to dislodge some of the metal
atoms from the cathode surface and produce an atomic cloud.
o A portion of sputtered metal atoms is in excited states and thus emits their
characteristic radiation as they return to the ground sate
o Eventually, the metal atoms diffuse back to the cathode surface or to the glass
walls of the tube and are re-deposited
82
83
Hollow cathode lamp (Radiation source):

 It should emit stable intense radiation of the element to be

determined, resonance line of the element.
 DC voltage is applied across anode and cathode
 Atom of the filler gas ionized at the anode and attracted by the
cathode
 Inert gas ions strike the cathode and displace the surface metal
atoms in the inert gas
 Further collision of vaporized metal atoms with energetic inert gas
ions result in excited metal atoms emit the characteristic spectrum of
the metal used to construct the cathode

 Each HCL emits the spectrum of the metal used in the cathode
 Spectral lines emitted by the copper HCL can only be absorbed by
the copper atom present in the flame. Hence its element specific.

84
Atomic Absorption Spectroscopy
Hollow Cathode Lamp (HCL)

?
 Instrumentation
(premix or laminar flow
burner)

Light separation and detection

• AA spectrometers use monochromators and detectors for uv and visible light.
• The main purpose of the monochromator is to isolate the absorption line.
• Photo multiplier tubes are the most common detectors for AA spectroscopy
86
 Principle
Absorption of energy by ground state atoms
present in the gaseous state in flame

N*/N0 = A e-DE / kT

Receive same l which they emit in the excited state

Absorption a density of atoms in flame

Amount of light absorbed a 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 a No. of Absorption intensity a 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