Chapter 24

Introduction to Spectrochemical
Methods

DEFINITION:
Spectroscopy- the interactions of radiation
and matter.
Spectroscopic Analytical Method- method
that based on measuring the amount of
radiation produced or absorbed by molecular
or atomic species of interest.
Electromagnetic Radiation- a form of energy
with properties that can described in terms of
waves or as particulate photons.

Properties of Electromagnetic Radiation

It can be described as a wave with properties of
wavelength, frequency, velocity, and amplitude.
In contrast to sound waves, it requires no
supporting medium for its transmission.
It can travels nearly a million times faster than
sound.
It can be treated as discrete packets of energy or
particles called photon or quanta.
A photon is directly proportional to its
frequency.

Wave Properties
Electromagnetic radiation is modelled as
waves consisting of perpendicularly oscillating
electric and magnetic fields

Amplitude, A : the vertical distance from the midline of awave to

the peak or through.
Period, : the time in second that it takes for complete a cycle.
Frequency, : the number of oscillations that occur in 1 second.
Wavelength, : the linear distance between successive maxima or
minima of awave.

Multiplication of the frequency (in waves per unit time) by the

wavelength (in distance per wave) gives the velocity (cm s-1, or ms-1).
Velocity,

Frequency,

Wavelength,

*In vacuum

Wavenumbers, cm-1

The velocity of light in vacuum:

c = v = 2.99792.00 108ms-1 = 3.00 108ms-1 = 3.00 1010cms-1

In a medium, light travels with a velocity less than c because of

interaction between the electromagnetic field and electrons in the
atoms or the medium.
The velocity, wavelength and intensity of the radiation will change as
radiation passes through different media, but the frequency of the
radiation is constant.

Example 24.1
Calculate the wave number of a beam of infrared
radiation with a wave length of 5.00 m.

Particle Properties of Electromagnetic Radiation

Sample which absorbs electromagnetic radiation will undergoes

a changes in energy.
Radiation is consist of photons or quanta (energetic particles).
Photon = A particle of light carrying an amount of energy equal
to h
We can relate the energy of a photon to its wavelength,
frequency and wave number by;
E h

hc
hc

h is Plancks constant (6.626 10-34 Js)

Example 24.2
Calculate the energy in jouls of one proton of
radiation described in Ex: 24.1, applying eq. E= hc/

Electromagnetic Spectrum;
The division of electromagnetic radiation on the basis of photons
energy.

Measuring Photons as a Signal

* Spectroscopy is possible if the photons interaction with the sample leads to a
change in one or more characteristic properties of electromagnetic radiation.

Absorption: The energy carried by a photon is absorbed by the analyte

, promoting the analyte from a lower-energy state to a higher
-energy or excited state.

Simplified energy level diagram showing absorption of a photon

Emission;
- The release of a photon when an analyte returns to a lower-energy
state from a higher-energy state.

Simplified energy level diagram showing emission of a photon

Absorbance;
- the attenuation of photons as they pass through a sample (A)
Absorbance spectrum;
- A graph of a samples absorbance of electromagnetic radiation
versus wavelength (or frequency or wavenumber)

*UV-Vis absorption spectrum

From bromotymol blue

The higher-energy state can be achieved in

several ways:
Thermal energy (flames and plasmas)
Radiant energy from a photon
Chemical reaction (exothermic reaction)

Measuring the electromagnetic radiation emitted as it

returns to the ground state.

The analyte is stimulated by heat or by electrical energy is called emission

spectroscopy.
The analyte is stimulated by a chemical energy is called chemicaluminescense
spectroscopy.

Measuring the emission of protons after absorption. It is called

photoluminescence spectroscopy.

The analyte is stimulated by an external electromagnetic radiation source.

The most important forms of photoluminescence for analytical purposes are
fluorescence and phosphorescence spectroscopy.
The major distinction between fluorescence and phosphorescence is the time
scale of emission, fluorescence being prompt and phosphorescence being
delayed.

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Transmittance and Absorbance

Transmittance: The ratio of the radiant power passing through a sample
to that from radiation s source.
T

P
P

T
0

PT: electromagnetic radiations power exiting from the sample

P0: electromagnetic radiations power that incident on the sample
from source
% Transmittance: multiplying by 100

Absorption of the radiation thus attenuates the beam depends on:

- Concentration of the absorbing molecules (concentration ,
attenuation ).
- The path length of the absorbing medium, (length , attenuation ).
The radiant power (watt) of the beam decreases from P to P.

In measuring transmittance and absorbance, we cannot use equation

A = log P
P
This is because reflection and scattering losses can occur at the cell walls.
To gain right calculation, the power of beam transmitted through a cell containing the
analyte solution is compared with an identical cell containing only the solvent or
reagent blank:
A = log P log P solvent
P
P solution

Alternative: absorbance

P
P
A log T log log
P
P
T

Absorbance and Concentration: Beers Law

P
P
A log T log log abC bC
P
P
T

a = constant, called the analytes absorptivity (cm-1 conc-1)

b = samples overall thickness (cm).
C = concentration of the absorbing species.
= constant, called the molar absorptivity (using when concentration
in moles per liter or M) (cm-1 M-1)
a and must have unit that cancel the units of b and c.

For multicomponent samples;

A A bC
m

i 1

i 1

* There are no interactions between the components.

Beers Law: The relationship between a samples absorbance

and the concentration of the absorbing species
*Calibration curves based on Beers Law are used routinely
in quantitative analysis.

Example 24.3
p.723

Absorption spectra

Absorption spectra
Atomic absorption
Molecular absorption
Limitations to Beer's law
Instrumental deviation

Atomic absorption

When a beam of polychromatic ultraviolet or

visible radiation passes medium contain gaseous
atoms.

Only few frequencies are attenuated by

absorption.
when recorded on a very high resolution
spectrometer, the spectrum consists of a number
of very narrow absorption lines.

Example 24.4
p.726

Molecular absorption

Undergo 3 types of electronic transition-(transfer of a

electron from one electronic orbital to another )
1. electronic
2. vibration
3. rotational

E = Eelectronic + Evibrational + Erotational

 Eelectronic associate with electrons in the various

outer orbital of the molecules.
Evirational inter atom vibrations
Erotational rotation of the molecule about its center of
gravity

Infrared spectra for molecules and

polyatomic ions
Infrared absorption
not sufficiently energetic to cause electronic
transitions.
it can induce transitions in the vibrational and
rotational states associated with ground electronic
state of molecule.

Ultraviolet and visible for molecules and ions

More energetic
Consists of absorption band- closely space line.
Typical absorption band consists of a large number of lines.
There are 3 different spectra according to the phase of the
species.
1. gas phase.
2. liquid phase and,
3. In aqueous solution.

Energy diagram showing

Difference between the
Absorption of IR (left) and
UV-Vis radiation (right)

Limitation to beers law

There are 3 exceptions in beers law

Fundamental limitations fundamental and represent real limitation to the law
i. too high concentration of the analyte.
ii. Dependence of the molar absorptivity upon the refractive index of the medium
* using low concentration of analyte
Chemical limitations chemical change that occur when the concentration changes.
i. Chemical reaction of analyte with solvent
* Same wavelength or use buffer system (to maintain pH)
Instrumental limitations method to make absorbance measurements
i. Polychromatic radiation give negative deviation (beers law valid for purely
monochromatic radiation only one wavelength)
ii. Stray radiation

Calibration curves showing positive and negative deviations from beers law

Application of Beers Law

Calculate molar absorptivity of Tryptophan (comp. of proteins) if the
radiation @280nm, through 1mm path length, the aqueous solution
concentration (0.5 mmol L-1 ), and only 54% of light passes through.

A= - log T= bC
= - log T/bC = log 0.54/ 0.5 X 10-3 mol L-1 X 1mm
= 5.4 X 10

L mol-1 mm

-1

Cont
What is the Absorbance for 1 mm and 5 mm?
For 1mm:
A= - log T = - log 0.54 = 0.27
For 5 mm:
A = b C = 5.4 X 10 2 L mol-1 mm -1 X 5mm X
0.5 X 10 -3 molL-1
= 1.35

Chemical deviation
Deviations from Beers Law appear when the
absorbing species undergoes association, dissociation,
or reaction with the solvent to give products that absorb
differently from the analyte (Example 24.5).
It can predicted from the molar absorptivities of the
absorbing species and the equilibrium constants for the
equilibria involved.

Instrument deviations:
Polychromatic radiation: it have continuous distribution of wavelengths used
with filters to isolate a nearly symmetric band of wavelengths a round the
wavelength to be employed (see p. 751, Section 25A-3 Wavelength selector).
Beers Law strictly applies only when measurements are made with
monochromatic radiation

A (measured) = bc = bc (p. 731 & 732)

Stray radiation:
- it is the radiation from the instrument that is outside the nominal wavelength
band chosen for the determination.
- It is often the result of scattering and reflection of the surface of lenses or
mirrors, filters, and windows. It can measured by:

A = log ( P0 + Pstray ) / ( PT + Pstray )

IR, UV-Vis
Spectrophotometry

Instrumentation for UV-Vis:

Filter photometer: A simple instrument for measuring absorbance that
uses absorption or interference filters to select the wavelength.

Spectrophotometer: An instrument for measuring absorbance that uses a

monochromator to select the wavelength.

Single-beam spectrophotometer (fixed-wavelength = monochromator)

Double-beam spectrophotometer

strumentation for IR:

Filter photometer

double-beam spectrometer

Fourier transform IR (using interferometer instead

f monochromator)

iv. Attenuate total reflectance (ATR)

v. Diffuse reflectance

Quantitative Applications
Usually using UV-Vis
IR can be used
- Environmental applications; analysis waters & waste waters
- Clinical applications: analysis of glucose
- Industrial analysis; analysis of iron content in food
- Forensic applications: Determination of blood alcohol

Atomic Absorption
Spectroscopy
(AAS)
Chapter 28 (p. 858)

Instrumentation:
- single-beam optics
- double-beam optics
Atomization:
the process of converting an analyte into a free atom.
(solid, liquid or solution into gaseous)
1. Flame atomizer
2. Electro thermal atomizers

Flame atomizer

Electrothermal atomizer/ graphic furnace

Electrothermal atomizers offer the advantage of unusually high
sensitivity for small volume of samples.
* Important to increase sensitivity (detection limits in picogram range)

Flame Vs Electrothermal atomization

- Concentration (diluted, concentrated)
- interference (high, low)
- volume of samples (high, low)

Quantitative Applications
* Analysis of trace metals

Preparing the Sample

- Sample in liquid or solution form

Minimizing Interference (p. 856)

i.

spectral interference
- Occur when an analytes absorption line overlaps with an
interferents absorption line or band.
- occur when components of the samples matrix react in the flame to
form molecular species such as oxides and hydroxides.
- Light scatter or absorption by solid particles, unvaporized solvent
droplets, or molecular species in the flame will cause a positive
interference in atomic absorption spectrophotometry. This is
especially a problem for wavelengths less than 300 nm, when
solutions of high salt content are aspirated because the salt may not
be completely desolvated or its molecules dissociated into atoms.

How to minimize:
i.

Analyzing a blank

ii. Use identical sample matrix and standard matrix. (avoiding interference
due to sample matrix and standard matrix by using identical matrix)
iii. interference due to samples matrix can be eliminated by adjusting the
flame composition.
iv. When the identity of sample matrix interferences is unknown
use background correction (use a continuum source)
v. Zeeman effect background correction
vi. Smith-Hieftje background correction
p.862

Minimize chemical interference

Formation of nonvolatile compounds often can be minimized by

ncreasing the temperature of the flame by changing the fuel-to-oxidant
atio or by switching to a different combination of fuel and oxidant.

dd releasing agent or protecting agent to solutions containing the analyte

effect of ionization can be minimized by adding a high concentration of an

nization suppressor

Standardizing the Method

-Linear calibration curve
- Usually using external standard
- if using electrochemical atomization, Standard additions is often used.

Spectroscopy Based on Emission

(Atomic Emission Spectroscopy)

Atomic Emission Spectra

- Occurs when a valence electron in a higher-energy atomic orbital returns
to a lower-energy atomic orbital.
- Intensity of emission line is proportional to the number of atoms.
- Atomization and excitation sources;

Flame Sources
Plasma Sources
Arcs
Sparks
Lasers

Plasma Sources
- Higher temperatures than flame
- Better atomization
- highly populated excited states
- better capability for multi-elemental analysis
- used rapid scanning monochromator or
multichannel instrument that allows for
the simultaneous monitoring of many analytes.
- Better detection limit than flame emission.
- Fewer spectral and chemical interferences

Diagram of inductive coupled plasma

Torch. (p.846)

Minimizing Interference for atomic emission

spectroscopy
i. Spectral interference: occurs due to the molecular emission.
How to minimize?
- Background corrections for flame emission are made by scanning over
the emission line and drawing baseline
It is less problematic for plasma emission

Method for background correction

in flame atomic emission.

ii. Minimize chemical interference

- Flame emission is subject to the same types of chemical interferences as
AAS.
- minimize,
*adjusting flame composition
*adding protecting agent
*adding releasing agent
*adding ionization suppressors
- with plasma source, chemical interferences are insignificant

Standardizing the Method

- For flame source, Linear calibration curve over two to
three orders of magnitude
- For plasma source, Linear calibration curve over four
to five orders of magnitude
-Quantitative analyses are best conducted using external
standard