Principles of

Spectrophotometric
Analytical Method
Session 2
Department of Chemistry
Universitas Indonesia

1
Introduction to Spectrometry methods

Spectroscopy:
Study of interaction between radiation (or other
forms of energy) and matter (a branch of
science).
Spectrometry:
Analytical methods based on atomic and
molecular spectroscopy

2
 Types of Analytical
Spectroscopy
 Absorption
 Fluoresence and Phosphoresence
 Emission (atomic with flames, arcs, sparks,
and palsmas)
 Chemilumenesence and Biolumenesence
 Reflection

3
 Kinds of Spectroscopy

Douglas A. Skoog, et al. Principles of Instrumental Analysis, Thomson, 2007

4
 LIGHT

Electro-magnetic
radiation

5
 Light as a Wave

Douglas A. Skoog, et al. Principles of Instrumental Analysis, Thomson, 2007 6

 Light as a Wave

Douglas A. Skoog, et al. Principles of Instrumental Analysis, Thomson, 2007 7

 Light as a Wave

Frequency = n
Velocity of propagation = v = nl
Speed of light in a vacuum = c = 3.00 x 108 m/s
Wavenumber (reciprocal of l) = n = kn = n/v

Douglas A. Skoog, et al. Principles of Instrumental Analysis, Thomson, 2007 8

 Effect of the Medium on a Light Wave

• Frequency remains the same.

• Velocity and Wavelength change.

Douglas A. Skoog, et al. Principles of Instrumental Analysis, Thomson, 2007 9

 Mathematic Description of a Wave

Y = A sin(wt + f)

A = Amplitude

f = phase angle

2pv
w = angular frequency = 2pn =
l
Y = A sin(2pnt + f)
Douglas A. Skoog, et al. Principles of Instrumental Analysis, Thomson, 2007 10
 Mathematic Description of a Wave

Sine waves with different amplitudes and with a phase different of 90 degree

Douglas A. Skoog, et al. Principles of Instrumental Analysis, Thomson, 2007 11

 Superposition of Waves
If two plane-polarized waves overlap in space, the
resulting electromagnetic disturbance is the algebraic
sum of the two waves.

Y = A1sin(2pn1t + f1) + A2sin(2pn2t + f2) +…….

Optical Interference: The interaction of two or more

light waves yielding an irradiance that is not equal to
the sum of the irradiances.

Coherence: When two waves have an initial phase

difference of zero or it is constant for a long time they
are considered coherent.
12
A ray of white-wavelength incident on a prism

R
B

White light

Cai® 2007 13
14
 Light as Particles

Eugene Hecht, Optics, Addison-Wesley, Reading, MA, 1998.

hc
E  hn 
l
h = Planck Constant = 6.63  10-34 Js
15
 The Photoelectric Effect
Vo: Stopping voltage
(the negative voltage at
which the photocurrent
is zero)

eV0 = hn - w

w: work needed to
remove e-

Douglas A. Skoog, et al. Principles of Instrumental Analysis, Thomson, 2007 16

  Current is proportional
to the intensity of the
radiation
 V0 depends on the
Cut-off n
frequency of the
radiation and the
chemical composition
of the coating
 V0 independent of the
intensity of the
radiation

Douglas A. Skoog, et al. Principles of Instrumental Analysis, Thomson, 2007 17

 Energy States of Chemical
 Quantum theory by  E1-E0 = hn = hc/l
Planck (1900)
 Black body radiation
 Atoms, ions , and
molecules exist in discrete
states
 Characterized by definite
amounts of energy
 Changes of state involve
absorption or emission of
energy

18
 Interaction of Radiation and Matter

Emission and
Chemiluminescence
Process
19
Interaction of Radiation and Matter

Absorption
Process

20
Interaction of Radiation and Matter

Photoluminescence method
(Fluorescence and
phosphorescence)

21
 Emission of Radiation

Excitation needs energy!

 Emission
•Particle bombardment (e-)
 X*  X + hn
•Electrical currents (V)
•Fluorescence
•Heat

Douglas A. Skoog, F. James Holler and Timothy A. Nieman, Principles of

Instrumental Analysis, Saunders College Publishing, Philadelphia, 1998.

22
Emission: Saltwater in a flame

23
 Line Spectra

Individual atoms, well separated, in a gas phase 24

 Band Spectra

Vibrational levels

Small molecules and radicals 25

 Continuum Spectra
 Produced when solid are heated to incandescence.
 Blackbody Radiation (Thermal Radiation)

26
 Blackbody Radiation
 A blackbody is a theoretical object, (i.e.
emissivity = 1.0), which is both a perfect
absorber and emitter of radiation.
Common usage refers to a source of
infrared energy as a "blackbody" when
it's emissivity approaches 1.0 (usually e
= 0.99 or better) and as a "graybody" if
it has lower emissivity.
 Important sources of infrared, visible,
and long wavelength UV for analytical
instruments
http://www.electro-optical.com/bb_rad/bb_rad.htm

27
 Blackbody Radiation

Wien’s
Displacement Law
2.897  10 6 K nm
l max 
T

Stefan-Boltzman Law
P = sT4
s = 5.6697  10-12 Wcm-2K-4

Both lmax and radiation power (P) are related to TEMPERATURE and current!

Eugene Hecht, Optics, Addison-Wesley, Reading, MA, 1998. 28

Continuum Source Line Source

Al + Mg

Continuum + Line Source

Ingle and Crouch, Spectrochemical Analysis 29

l Ranges of Common Sources

Douglas A. Skoog and James J. Leary, Principles of Instrumental

Analysis, Saunders College Publishing, Fort Worth, 1992. 30
Optical Source Characteristics

Ingle and Crouch, Spectrochemical Analysis

31
 Nernst Glower
Rare earth oxides formed into a
cylinder (1-2 mm diameter,
~20mm long).
Ingle and Crouch, Spectrochemical
Pass current to give: Analysis
T = 1200 – 2200 K.

Douglas A. Skoog and James J. Leary, Principles of Instrumental

32
Analysis, Saunders College Publishing, Fort Worth, 1992.
 Globar
Silicon Carbide Rod (5mm diameter, 50 mm long).
Heated electrically to 1300 – 1500 K.
Positive temperature coefficient of resistance
Electrical contact must be water cooled to prevent arcing.

33
Ingle and Crouch, Spectrochemical Analysis
 Tungsten Filament
Heated to 2870 K.
Useful Range: 350 – 2500nm

Ingle and Crouch, Spectrochemical Analysis 34

 Tungsten / Halogen
Iodine added.
Reacts with gaseous W near the quartz wall to form WI2.
W is redeposited on the filament.
Gives longer lifetimes
Allows higher temperatures (~3500 K).

35
Intensity Spectrum of the
Tungsten-Halogen Lamp

• Weak intensity in
UV range
• Good intensity in
visible range
• Very low noise
• Low drift

36
 Arc Lamps

Electrical discharge is
sustained through a gas or
metal vapor.

Continuous emission due to

rotational/vibrational energy
levels and pressure
broadening.

37
Ingle and Crouch, Spectrochemical Analysis
 H2 or D2 Arc Lamps
D2 + Ee-  D2*  D’ + D” + hn

Energetics: Ee- = ED2* = ED’ + ED” + hn

Useful Range: 185 – 400 nm.

38
Ingle and Crouch, Spectrochemical Analysis
Intensity Spectrum of the
Xenon Lamp

• High intensity
in UV range
• High intensity
in visible range
• Medium noise

39
 Hg Arc Lamp
Continuum + Line Source

High Power Source.

Often used in photoluminescence.

40
Ingle and Crouch, Spectrochemical Analysis
 Hollow Cathode Discharge Tube.
Apply ~300 V across
electrodes.
Ar+ or Ne+ travel toward the
cathode.
If potential is high enough
cations will sputter metal off
the electrode.
Metal emits photons at
characteristic atomic lines as
the metal returns to the
ground state.

Douglas A. Skoog and James J. Leary, Principles of Instrumental 41

Analysis, Saunders College Publishing, Fort Worth, 1992.
Hollow Cathode Discharge Tube.

Line Widths are typically 0.01 – 0.02 Å.

42
Ingle and Crouch, Spectrochemical Analysis
 Absorption of Radiation
 Is a quantized process???
 The energy absorbed is released, although not necessarily
all as light energy (e.g. heat)
 Results in excitation of a molecule to a higher energy
state
 E= E electronic + E vibrational + E rotational

43
 Absorption of Radiation

Douglas A. Skoog, F. James Holler and Timothy A. Nieman, Principles of

44
Instrumental Analysis, Saunders College Publishing, Philadelphia, 1998.
Atomic absorption

45
Rotational energy levels associated with each
vibrational level not shown
46
 Relaxation
 Resonance fluorescence
 lF = lA
 Non- Resonance fluorescence
 lF  lA
 Stokes shift
 lF > lA

47
 Quantitative Aspects of
Spectrochemical Measurements
 Radiation power P
 The energy of the a beam of radiation that
reaches a given area per second
 Radiation power is determined with a radiation
detectorthat convert radiation energy into
electrical signal
 S =kP

S is an electrical signal, generally S is a

voltage or current that proportional to radiation
power, whereas k is contant
 Dark current
 Response of the detector in the absence of
radiation, kd
48
 S =kP + k
 Quantitative Aspects of
Spectrochemical Measurements
 Transmittance
 T = P/Po (definition)
 Po - incident light power
 P - transmitted light power
 %T = P/Po x 100 %
 Absorbance
 A = - log T (definition)
 Beer’s Law (physical law applicable under
certain conditions)
 A =  b c (basis of quantitation)
  - molar absorptivity (L mol-1 cm-1)
 b - pathlength (cm)
 c - concentration (mol L-1)
49
 Non-radiative relaxation
 Vibrational Relaxation:
 A molecule can give off some of its energy from absorbed light
(usually uv-vis) by jumping to a lower energy vibrational state.
 The excess energy is used to make the conversion. No light is given
off.
 Internal Conversion:
 The molecule transitions to a lower energy electronic state without
giving off light.
 Excess energy is used to covert the molecule from one electronic state
to another.
 Poorly understood
 External conversion:
 The molecule gives off energy to an external source, such as by
collision with another similar molecule or solvent molecule. This is
called “quenching”
 Intersystem Crossing:
 The molecule goes from a singlet to triplet excited state and uses
up energy changing the spin of an electron.
50