Course Name: Spectrophotometric and Electroanalytical Methods

Course Code: CHMI2203 & CHM2203

Lecture 2 contents
• Components of Optical Instruments
 Components of Optical Instruments

The essential components of a spectrophotometer include:

1- A stable source of radiation

2- A wavelength selector
3- A transparent container to hold the sample
4- Radiation detector
5- Readout devices (computer)
 Information

• Visible wavelengths cover a range

from approximately 400 to 800 nm.

• UV wavelengths cover a range from

approximately 100-400 nm
 Components of Optical Instruments

The following scheme represent most instrumental setup:

Radiation Wavelength Sample Detector Processor

Source Selector Cell
 Components of Optical Instruments

1- Sources of Radiation
A source to be used in a selected range of wavelength should
have the following properties:
It should generate a beam of radiation covering the wavelength range
in which to be used. For example, a source to be used in the visible region
should generate light in the whole visible region (340-780 nm).

The radiation of the source should be stable and intense for reasonable
periods of time .
 Components of Optical Instruments

Type of sources:
A- continuous sources
Emit radiation of all wavelengths within the spectral region for
which they are to be used .

Continuous source

Visible and near IR radiation Ultraviolet radiation

tungsten lamp Deuterium and hydrogen lamp

340-2500 nm 180-350 nm
 Components of Optical Instruments
Type of sources:
B- Line sources
A line source is a source which has a line output at definite wavelengths, rather
than arrange of wavelengths. Hollow cathode and electrode less discharge lamps
are examples of line sources which produce few sharp lines in the UV and visible
(Vis).
 Components of Optical Instruments

2 - Wavelength Selectors
These are devices which resolve wide band polychromatic
radiation from the source into a narrow band that is suitable
for analyte. (function)
advantage :These wavelength selectors greatly enhance
both the selectivity and the sensitivity of an instrument and
can be reduce the deviations of Beer's law.
 Components of Optical Instruments
Type of Wavelength Selector

Wavelength Selector

Filters Monochromators

prism grating
monochromator monochromator
 Components of Optical Instruments
Type of Wavelength Selector

a. Filters prepared from special materials allow transmission of only limited wavelength
regions while absorbing most of the radiation of other wavelengths.

b. Monochromators: resolve polychromatic into its individual wavelengths and

isolate these wavelengths into very narrow bands.
grating monochromator
 Components of Optical Instruments

3- Sample Containers

The cell or cuvette is a small tube of circular or square cross section,

sealed at one end, made of plastic, glass, or fused quartz and
designed to hold samples for spectroscopic experiments.

Crystalline
Quartz Glass sodium crystal
• UV region • visible • IR region
• Visible region.
region

Why isn't glass used in UV spectroscopy?

 Components of Optical Instruments
Continue Sample Containers
 Components of Optical Instruments

4-Detectors
The detectors are devices that convert radiant energy
into electrical signal (function).
Criteria or requirements

A Detector should be sensitive, and has a fast

response over a considerable range of wavelengths.

In addition, the electrical signal produced by the

detector must be directly proportional to the
transmitted intensity (linear response).
 Components of Optical Instruments
4-Detectors
There are two broad classes of spectroscopic transducers:
a. photon transducers .
b. thermal transducers.

Examples of transducers for spectroscopy

 Components of Optical Instruments

Continue on detectors
a. Photon detectors
Phototubes and photomultipliers tube contain a photosensitive
surface that absorbs radiation in the ultraviolet, visible, or near
IR, producing an electrical current proportional to the number of
photons reaching the transducer.

b. Thermal detector
Use for IR spectroscopy.
 Components of Optical Instruments
continue on detectors
Phototube
Phototube emits electrons from a photosensitive, negatively
charged cathode when struck by visible or UV radiation
The electrons flow through vacuum to an anode to produce
current which is proportional to radiation intensity.
 Components of Optical Instruments
continue on detectors

The photomultipIier tube

Process:
a) light hits photoemissive cathode and e- is emitted.
b) an emitted e- is attracted to electrode #1
(dynode 1), which is 90V more positive.
Causes several more e- to be emitted.
c) these e- are attracted to dynode 2, which is
90V more positive then dynode 1, emitting
more e-.
d) process continues until e- are collected at
anode after amplification at 9 dynodes.
e) overall voltage between anode and cathode
is 900V.
f) one photon produces 106 – 107 electrons.
g) current is amplified and measured.
 Types of UV-visible instruments

There are two types of instrument:

a. Single beam instruments
b. Double beam instruments

The blank:
The blank contains all substances expect the analyte.
Is used to set the absorbance to zero:
Ablank = 0
 Important concepts
Absorption spectrum: a plot of the amount of light absorbed by a
sample ( atom or molecule ) as a function of wavelength.

ato
m

molecu
le
 Absorption and emission of EMR
Electron Energy level E
2

Excited
state
E Electron energy
1 level

Vibrational
energy level
Rotational
energy level

Ground E
0
state
 Atomic Absorption
• When a EMR (UV or Vis) pass through a sample containing
monoatomic particles (e.g.,Na) it results in the absorption
of a well-defined frequencies (wavelengths).
• relative simplicity of spectra( line spectra) is due to small
number of possible energy levels (just electronic states ,
no rotational or vibrational sates).
 Atomic absorption spectrum
Na vapors exhibits two sharp absorption peaks (589.0
and 589.6nm) due to excitation of 3s electron to two
3p orbitals.
 Molecular Absorption
Absorption spectra of molecules are more complex than
atomic spectra because the number of energy states of
molecules is enormous (electronic , vibrational and rotational
states ).

Molecular Absorption
spectrum
 Important concepts

Emission spectrum: a plot of intensity of

emitted radiation as a function of wavelength
 Types of UV-visible instruments

Single beam spectrophotometer :

It consists of the radiation source, a filter or monochromator for

wavelength selection, matched cells that can be imposed alternately
in the radiation beam, the photodetector, an amplifier and readout
device.
 Types of UV-visible instruments
Double beam spectrophotometer:
Two beams are formed in space by a V-shaped mirror called a beam splitter.
One beam passes through a reference solution to a detector, and the second
simultaneously traverses the sample to a second, matched detector. The two
outputs are amplified, and their ratio (or the log of their ratio) is determined
electronically and displayed by the readout device.
Double beam spectrophotometer
 Types of UV-visible instruments

Advantages of Single Beam Systems

less expensive
Disadvantages
The sample and blank must be placed alternately in the light
path.
For measurements at multiple wavelengths, the blank must
be run at each wavelength.
 Types of UV-visible instruments

Advantages of Double Beam Systems

The absorption of the sample is automatically corrected for
the blank absorption , since the detector reads the
difference between the sample beam and the blank beam.
More accurate.