SUBTOPICS

BASIC INSTRUMENTAL ANALYSIS

Spectroscopy
CHM260


 Properties of electromagnetic radiations

 The electromagnetic spectrum

CHAPTER 1  Spectroscopic measurements

AN INTRODUCTION TO  Absorption methods

 Beer’s law
SPECTROSCOPIC  Interaction of electromagnetic radiation

METHODS OF with matter

 Absorption spectra
ANALYSIS  Emission of electromagnetic radiation

SPECTROSCOPY
 Spectroscopy: The study of the interaction
SPECTROSCOPIC
between ELECTROMAGNETIC (EM)
RADIATION and MATTER.
ANALYSIS
 The sample solution absorbs EM from an covers
appropriate source, and the amount absorbed
is related to the concentration of the analyte
in the solution.
 Spectrophotometer/Spectrometer: The
instrument that determines the absorption
ATOMIC MOLECULAR
spectrum. SPECTROSCOPY SPECTROSCOPY
 PROPERTIES OF THE ELECTROMAGNETIC RADIATIONS
 Electromagneticradiation is the emission
and transmission of energy in the form of
electromagnetic waves.
 EM shows wave like properties and
particle like properties (photon or
quanta) at the same time.
WAVE PROPERTIES
 EM radiation is conveniently modeled as waves
consisting of perpendicularly oscillating electric and
magnetic fields, as shown in the diagram.
 At 90° to the direction of propagation is an oscillation in
the ELECTRIC FIELD.
 At 90° to the direction of propagation and 90° from the
electric field oscillation (orthogonal) is MAGNETIC
FIELD oscillation.
I. WAVE-LIKE PROPERTIES
 Energy radiated in the form of a WAVE caused
by an electric field interacting with a magnetic
field.
 Result of the acceleration of a charged particle.
 Does not require a material medium and can
travel through a vacuum.
• Electric field
(red color)
• Magnetic field
(blue color)
WAVE PARAMETERS
 Period (p) – the time required for one cycle to pass a
fixed point in space.
 Frequency (v) – the number of cycles which pass a fixed
point in space per second.
v = 1/p (s-1 = Hz)
v depends on the source, but is independent of the
propagating (transmitting) material.
 Amplitude (A) – The maximum length of the electric
vector in the wave (maximum height of a wave).
 Wavelength (λ) – The distance between two identical
adjacent points in a wave (usually maxima or minima).
  Velocity of propagation (vi)
- The rate at which a wave front moves through a
propagating medium in meters per second (ms-1).
- how fast the wave is moving in the specific
medium.

vi = vλi

Period (p) Velocity of light (c)

c = 3.00  108 ms-1
= 3.00  1010 cms-1

c = vλ = 3.00  108 ms-1

 Wavenumber (v)  Intensity (I)

- The number of waves per cm in units of cm-1 - The radiant power emitted per unit solid angle.
v = 1 = v - Describes the radiant power of a source emitted in a
λ c certain direction.
- SI unit for intensity: watts per steradian (Wsr-1)
 Radiant Power (P) - Solid angle (Ω): the size of area divided by the square
- The amount of energy reaching a given area per second. of the radius (r2) of the sphere. Unit solid angle is
- Unit: watts (W). steradian (sr).
- Total power of radiation emitted by a source (lamp, light Ω = A/r2
emitting diode, etc.), transmitted through a surface or
impinging upon a surface.
 II. PARTICLE NATURE OF LIGHT: PHOTONS
 Wave theory failed to explain phenomena
associated with the absorption and emission of
radiation of radiant energy.
 Thus, EM is viewed as a stream of discrete
particles, or wave packets of energy called
photons.
 Energy of photon is related to the frequency,
wavelength and wavenumber by this equation:
E = hv = hc = hcv
λ
h = Planck’s constant = 6.63  10-34 J.s
c = 3.00  108 ms-1
Example:
If the wavelength used in an instrument is changed
from 460 nm to 560 nm:
i) has the energy been increased or decreased?
ii) has the frequency been increased or decreased?
iii) has the wavenumber been increased or decreased?
Solution:
(refer to formula)
E = hv = hc = hcv
λ
i) When  increase, energy will decreased.
ii) When  increase, frequency will decreased.
iii) When  increase, wavenumber will decreased.

THE ELECTROMAGNETIC SPECTRUM

 The electromagnetic spectrum is arbitrarily broken down
Regions of the UV, Visible and IR
into different regions according to the wavelength. Spectrum

Region Wavelength Range

UV 200 – 380 nm

Visible 380 – 780 nm

Near-IR 0.78 – 2.5 m

Mid-IR 2.5 – 15 m
 Types of spectroscopic methods based on EM
Wavelength Units for Various Spectral
radiation
Region

Region Unit Definition (m)

X-ray Angstrom unit, Å 10-10 m

Ultraviolet/ Nanometer, nm 10-9 m

visible
Infrared Micrometer, m 10-6 m

1 Å = 10-10 m = 10-8 cm
1 nm = 10-9 m = 10-7 cm
1 m (micron) = 10-6 m = 10-4 cm

QUESTIONS:
Consider an electromagnetic radiation with a ANSWERS:
frequency of 5.25 x 1013 Hz.
i)  = 5.71 m
i) Calculate the wavelength (in microns) of the ii) Region: Infrared
radiation.
iii)E = 3.48 x 10-20 J
ii) Name the region in which this
electromagnetic radiation is located.
iii) What is the energy (in Joules) of the
radiation?
 SPECTROSCOPIC MEASUREMENTS Color of Different Wavelength Regions
HOW DOES MATTER ABSORB RADIATION?
Wavelength Absorbed color Transmitted Color
 The absorption of radiation can be obtained by Absorbed (nm) /Observed Color
considering the absorption of light in the visible. (Complement)
 The color of an object we see is due to the 380-450 Violet Yellow-green
wavelengths transmitted or reflected. The other 450-495 Blue Yellow
wavelengths are absorbed.
 Polychromatic light (white light) is visible light 495-570 Green Violet
that contains all the colors of the rainbow. 570-590 Yellow Blue
 If white light is passed through an object, the
object will absorb certain of the wavelengths, 590-620 Orange Green-blue
leaving the unabsorbed wavelengths to be 620-750 Red Blue-green
transmitted (seen as a color with our eyes).

Solution of
KMnO4
ABSORPTION METHODS
ABSORPTION
 Example: METHODS
- A solution of KMnO4 absorbs light in the
green region of the spectrum with an
absorption maximum of 525 nm, and the
solution is purple/violet (observed color). TRANSMITTANCE (T) ABSORBANCE (A)
Wavelength Absorbed color Transmitted Color
Absorbed (nm) /Observed Color
(Complement)
380-450 Violet Yellow-green

450-495 Blue Yellow

495-570 Green Violet

570-590 Yellow Blue

590-620 Orange Green-blue

620-750 Red Blue-green

 TRANSMITTANCE (T) ABSORBANCE (A)
– The fraction of incident radiation (electromagnetic
- A measurement of the amount of radiant power
radiation) that passes through the sample medium.
absorbed by the sample defined as the negative log of
transmittance.
- Absorbance has a linear relationship or directly
proportional with sample concentration defined by
Beer’s Law.

A = Absorbance
b = path length

Commonly expressed as
a percentage:

% T = P x 100 % A = Absorbance
P0 b = path length

QUESTION 1: ANSWER:

Convert the following percent transmittance i) %T = 33.6

data into absorbance: T = 33.6 / 100
= 0.336
i) 33.6
ii) 92.1 A = - log T
iii) 1.75 = - log 0.336
= 0.474
Answer: i) 0.474
ii) 0.0357
iii) 1.76
QUESTION 2: BEER’S LAW
• For monochromatic radiation, absorbance is directly
Convert the following absorbance data into
proportional to the path length (b) through the medium
percent transmittance: and the concentration (c) of the absorbing species.
i) 0.375 • These relationships are given by this following
ii) 1.325 equation.
iii) 0.012 A = abc
Answer: Concentration
Absorbance (g/liter)
i) A = - log T
T = Antilog (-A) %T = 0.4212 x 100 Absorptivity Pathlength
(Lg-1cm-1) (cm)
= Antilog (-0.375) = 42.12%
-The term “a” is a proportionality constant called
= 0.4212 absorptivity.
ii) 4.73% - Absorptivity is a constant for a given chemical species
iii) 97.3% at a specific wavelength.
* Monochromatic: light of a narrow frequency.

BEER’S LAW EXAMPLES:

 When the concentration is expressed in moles per 1. What is the concentration of an absorbing
liter and cell length is in cm, the absorptivity is species if its molar absorptivity is 1500
called the molar absorptivity and the symbol is  L/mol.cm and the measured absorbance
and the unit for  is liter mol-1 cm-1 in a 1.00 cm cuvette is 0.742?

A = bc Answer:
A = bc
 = molar absorptivity, liter mol-1 cm-1 c = A / b
b = sample path length, cm c= 0.742
c = concentration, mol per liter (1 cm)(1500 L/mol.cm)
= 4.95  10-4 M
 3. A sample in a 1.0 cm cell is determined with a
2. The measured absorbance of a sample in a 1.00 cm spectrometer to transmit 80% light at a certain
cuvette is 0.544. If the concentration is 1.40  10-3 M, wavelength. If the absorptivity of this
what is the molar absorptivity for the species? substance at this wavelength is 2.0, what is the
concentration of the substance?
Answer:
Answer:
A = bc The percent transmittance is 80%. So, T=0.80
 = A / bc A = abc
= 0.544 - log T = 2.0 L/g.cm  1 cm  c
(1 cm)(1.40  10-3 mol/L)
- log 0.80 = 2.0 L/g  c
= 389 L/mol.cm
c = 0.0969
2.0 L/g
= 0.0485 g/L

PARAMETERS THAT AFFECT ABSORBANCE

A = abc

 Concentration, c
 Width of cuvette, b

 Inherent ability for absorbing species

to absorb light
 i) Width of cuvette, b: APPLICATION OF BEER’S LAW
- Wider cuvette consist
of more absorbing b  Applying Beer’s Law to Mixtures
species present in the
path of the light,
hence absorbance is - Beer’s law also applies to solutions containing
greater. more than one kind of absorbing substance.
b - The species do not interact.
- Total absorbance for a multicomponent system at a
ii) Inherent ability for the absorbing species to single wavelength is the sum of the individual
absorb light absorbances.
- Chemical species vary with respect to this
inherent ability since absorption depends on Atotal = A1 + A2 + …………+ An
individual electronic and vibrational transitions
available in a given species. = 1bc1 + 2bc2 + ……… + nbcn

LIMITATIONS TO THE APPLICABILITY OF 1. Real deviations

BEER’S LAW
 Deviations are frequently observed from the direct
proportionality between absorbance, A and
concentration, c when pathlength, b is constant.
 Deviations may be due to:
1. Real deviations
2. Instrumental deviations
3. Chemical deviations
- fundamental deviations due to the limitations of the
law itself.
- Beer’s law: the absorption behavior of media containing
low analyte concentration.
- At high concentration (>0.01M) the extent of solute-
solvent interactions, solute-solute interactions, or
hydrogen bonding can affect the analyte environment
and its absorptivity.
- At high concentration, the average distances between
the molecules and ion are diminished where each
particle affects the charge distribution of its neighbors.
- Therefore, this interaction alter the ability of analyte
species to absorb a given wavelength of radiation.
- Causing deviation from the linear relationship between
absorbance and concentration.
2. Instrumental deviations
a) Due to polychromatic radiation
- Polychromatic: describe light that exhibits more
than one color, which also means that it contains
radiation of more than one wavelength.
- Beer’s Law strictly applies when measurements are
made with monochromatic source radiation.
- In practice, however, it is common to use a
polychromatic source of radiation with continuous
distribution of wavelengths along with a filter or a
grating unit (monochromators) to create a
monochromatic beam from this source.
- Deviations occur if the radiation is polychromatic
since the relationship between A and c is no longer
linear when  is differ.
 Why absorption measurements are taken at wavelength
of maximum absorbance λmax?
- λmax = Wavelength at maximum absorbance
- If the band of wavelength selected on the
spectrometer is such that the molar absorptivities of
the analyte is essentially constant, deviations from
Beer’s law are minimal.
- However, if a band is chosen such that the molar
absorptivity of the analyte at these wavelengths
changes a lot, the absorbance of the analyte will not
follow Beer’s law.
- The deviations in absorbance over wavelengths is
minimal when the wavelength observed is at the λmax.
Due to this reason, absorption measurements are
taken at wavelengths.
b) Due to presence of stray radiation
- Due to instrument imperfections.
- Stray radiation or scattered radiation is defined as radiation
from the instrument that is outside the nominal wavelength
band selected.
- Usually the wavelength of the stray radiation is very
different from the wavelength band selected. It is known that
radiation exiting from a monochromator is often
contaminated with minute quantities of scattered or stray
radiation.
- This stray radiation is due to reflection and scattering by the
surfaces of lenses, mirrors, gratings, filters and windows.
- The wavelength of stray radiation differs greatly from the
principal radiation and may not have passed through the
sample.
- If the analyte absorbs at the wavelength of the stray
radiation, a deviation from Beer’s law is observed similar to
the deviation due to polychromatic radiation.
- When measurements are made in the presence of
stray radiation, the observed absorbance, A’ is
given by this equation.
A’ = log P0 + Ps
P + Ps
Ps - power of non-absorbed stray radiation
% stray radiation = Ps  100
P0
3. Chemical deviations
- Occur when the analyte undergo dissociation,
association or reaction with the solvent to give
products that absorb differently than the
analyte.
- Example: the ionisation of acidic or basic
indicator
c) Mismatched Cells
 If the cells holding the analyte and the blank
solutions are having different path-lengths, or
unequal optical characteristics, it is obvious that
there would be a deviation observed in Beer’s law.
 In such cases when a plot of absorbance versus
concentration is made, the curve will have an
intercept k and the equation will be defined as:
A = εbc + k
Example:
- Phenol red undergoes a
resonance transformation
when moving from the acidic
form (yellow) to the basic form
(red).
- Due to this resonance, the
electron distribution of the
bonds of molecule changes
with the pH of the solvent in
which it is dissolved.
- The absorption spectrum of
the sample changes with the
change in pH of the solvent.
  The interaction of radiation with matter can cause
redirection of the radiation and/or transitions between
the energy levels of the atoms or molecules.

INTERACTION OF ELECTROMAGNETIC  A transition from a lower level to a higher level with

transfer of energy from the radiation field to the atom
or molecule is called absorption.
RADIATION WITH MATTER  A transition from a higher level to a lower level is called
emission if energy is transferred to the radiation field,
or nonradiative decay if no radiation is emitted.

 Redirection of light due to its interaction with matter is

called scattering, and may or may not occur with
transfer of energy, i.e., the scattered radiation has a
slightly different or the same wavelength.

 When radiation pass through a layer of solid,

ABSORPTION SPECTRA liquid, or gas, certain frequencies may be
selectively removed by absorption (a process in
which electromagnetic energy is transferred to the
atoms, ions, or molecules composing the sample.
ABSORPTION
SPECTRA
 For absorption of radiation to occur, the energy of
the excited photon must exactly match the energy
difference between the ground state one of the
ATOMIC MOLECULAR excited states of the absorbing species.
ABSORPTION ABSORPTION

ATOMIC ABSORPTION
 The passage of polychromatic ultraviolet or visible radiation
through a medium that consist of monoatomic particles, such as
gaseous mercury or sodium, results in the absorption, but a few
well-defined frequencies.
 The spectra is simple due to the small number of possible
energy states for the absorbing particles.
 Atomic spectra: sharp, well defines lines
 Example:
- Sodium vapor exhibits two closely spaced, sharp absorption
peaks in yellow region of the visible spectrum (589.0 and 589.6
nm).
- Reason: Excitation of the 3s electron and two 3p states that
differ only slightly in energy.
Absorbance
MOLECULAR ABSORPTION
More complex than atomic spectra because
the number of energy states of molecules is
generally large when compared with the
number of energy states for isolated atoms.
Energy associated with the bands of a
molecule is made up of three components:
E = Eelectronic + Evibrational + E rotational
Spectra of benzene vapor:
- a series of closely spaced absorption lines:
rotational energy levels associated with each
vibration state.
Spectra of benzene in hexane:
- broad smooth peaks: using high-resolution

instrument
Spectra of biphenyl in hexane:
- continous spectra: in the condensed state
Wavelength, nm
and in the solvent molecules

PROCESS OF MOLECULAR TRANSITION ENERGY LEVEL DIAGRAM OF MOLECULAR TRANSITIONS

Process of molecule to absorb radiation.
i) Rotational Transition
- Molecule rotates about various axes.
- Erotation being at definite Elevels
- Therefore, the molecule absorb radiation and be raised to a
higher rotational energy level.
- Occurs at very long wavelengths
ii) Vibrational Transition
- Atoms or group of atoms within the molecules vibrate relative
to each other.
- Evibration occur at quantized levels
- Therefore, the molecule absorb radiation and be raised to a
higher vibrational energy level. Energy level:
Electronic > vibrational > rotational
iii) Electronic Transition
- Electron of a molecule may be raised to a higher electron • rotational transitions – microwave or far-infrared region
• vibrational transitions – near-infrared region
energy .
• electronic transitions – visible and ultraviolet regions
 EMISSION OF RADIATION
EMISSION OF ELECTROMAGNETIC RADIATION  Electromagnetic radiation is produced when excited
particles (atoms, ions, or molecules) relax to lower
energy levels by giving up their excess energy as
EMISSION OF photons.
ELECTROMAGNETIC
RADIATION
 Source of excitation:
i) bombardment with electrons or other elementary
LINE BAND CONTINUUM particles. Leads to the X-radiation.
SPECTRUM SPECTRUM SPECTRUM
ii) Exposure to an electric current, an ac spark, or an
intense heat source (flame, dc arc, or furnace). Produce
UV, visible or infrared radiation.
iii) Irradiation with a beam of electromagnetic
radiation. Produces fluorescence radiation.
iv) An exothermic chemical reaction that produces
chemiluminescence.

LINE SPECTRA
• When an atom changes energy
 Made up of a series of sharp, well-defined peaks. state, it absorbs or emits energy
equal to the energy difference
 Caused by excitation of individual atoms. ∆E = E1 – E0
 Atomic transitions are usually very discrete
• The wavelength or frequency of
changes of electrons from one quantum state to radiation absorbed or emitted
another energy level (shells, spins, etc). during a transition proportional
 Only electronic transition is quantized. to ∆E

 No vibrational or rotational transition. ∆E = hv = hc


• Produce line spectra

E0 – lowest energy electronic level or ground state

E1, E2 – higher-energy electronic levels
 BAND SPECTRA CONTINUUM SPECTRA
 Encountered in spectral sources  Continuum spectra: A beam of light that contains a broad,
when gaseous radicals or small smooth distribution of photon wavelengths.
molecules are present.  Continuum radiation is produced when solids are heated to
 Molecular transition consists of 3 incandescence.
processes:  Thermal radiation is called black body radiation.
i) Rotational transition
- is characteristic of the temperature of the emitting surface
ii) Vibrational transition rather than the material of the surface.
iii) Electronic transition
- produced by the innumerable atomic and molecular
∆E = ∆Eelectronic + ∆Evibrational + ∆E rotational oscillations excited in the condensed solid by the thermal
energy.
 Band spectra is produced due
to vibrational and rotational
transitions.
Energy

Emission spectrum of a brine sample obtained with an

oxyhydrogen flame

Consists of the
superimposed
line, band and
continuum
spectra as
constituents of
the sample
* Energy peaks shift to shorter
wavelengths with increasing
temperature
 Line spectra

Band spectra

Continuum spectra