Spectrophotometry. Principle and Applications

EUROPEAN UNION Structural Funds
GOVERNMENT OF ROMANIA GOVERNMENT OF THE REPUBLIC 2007 - 2013

OF SERBIA
SPECTROPHOTOMETRY. PRINCIPLE AND APPLICATIONS
As.dr.ing. ADRIAN EUGEN CIOABLA
WORKSHOP
05 – 06 September 2013, Zrenjanin, Republic of Serbia
In the frame of the project
Sustainable development of an research center in Banat region and

Danube flow area through scientific research and environmental
simulation tools to asses and evaluate potential threats
www.envirobanat.ro
In chemistry, spectrophotometry is the quantitative
measurement of the reflection or transmission properties of a
material as a function of wavelength.
Spectrophotometry involves the use of a spectrophotometer,
which is a photometer that can measure intensity as a function
of the light source wavelength.
A spectrophotometer is commonly used for the measurement of
transmittance or reflectance of solutions, transparent or opaque
solids, such as polished glass, or gases.
Historically, spectrophotometers use a monochromator
containing a diffraction grating to produce the analytical
spectrum, but most modern mid-infrared spectrophotometers
use a Fourier transform technique to acquire the spectral
information (infrared spectroscopy).
UV – VIS AND IR SPECTROSCOPY
Ultraviolet–visible spectroscopy or ultraviolet-visible spectrophotometry

(UV-Vis or UV/Vis) refers to absorption spectroscopy or reflectance
spectroscopy in the ultraviolet-visible spectral region. This means it uses
light in the visible and adjacent (near-UV and near-infrared (NIR))
ranges.
UV/Vis spectroscopy is routinely used in analytical chemistry for the

quantitative determination of different analytes, such as transition metal
ions, highly conjugated organic compounds, and biological
macromolecules. Spectroscopic analysis is commonly carried out in
solutions but solids and gases may also be studied.
The instrument used in ultraviolet-visible spectroscopy is called a UV/Vis
spectrophotometer. It measures the intensity of light passing through a
sample (I), and compares it to the intensity of light before it passes through
the sample (I0). The ratio is called the transmittance, and is usually
expressed as a percentage
UV – VIS AND IR SPECTROSCOPY
Infrared spectroscopy (IR spectroscopy) is the spectroscopy that deals with

the infrared region of the electromagnetic spectrum, that is light with a
longer wavelength and lower frequency than visible light. It covers a range
of techniques, mostly based on absorption spectroscopy. As with all
spectroscopic techniques, it can be used to identify and study chemicals. A
common laboratory instrument that uses this technique is a Fourier
transform infrared (FTIR) spectrometer.
The infrared spectrum of a sample is recorded by passing a beam of
infrared light through the sample. When the frequency of the IR is the
same as the vibrational frequency of a bond, absorption occurs.
APPLICATIONS
The use of spectrophotometers spans various scientific fields, such as
physics, materials science, chemistry, biochemistry, and molecular
biology.
There are two major classes of devices: single beam and double beam. A
double beam spectrophotometer compares the light intensity between two
light paths, one path containing a reference sample and the other the test
sample. A single-beam spectrophotometer measures the relative light
intensity of the beam before and after a test sample is inserted.
Single beam spectrophotometer http://en.wikipedia.org/wiki/Spectrophotometry

APPLICATIONS
Although comparison measurements from double-beam instruments are easier
and more stable, single-beam instruments can have a larger dynamic range
and are optically simpler and more compact. Additionally, some specialized
instruments, such as spectrophotometers built onto microscopes or telescopes,
are single-beam instruments due to practicality.
Double beam spectrometer

http://chemwiki.ucdavis.edu/Physical_Chemistry/Spectroscopy/Electronic_Spectroscopy/Electronic_Spectroscopy%3A_Application
APPLICATIONS
Infrared spectroscopy is a simple and reliable technique widely used in both

organic and inorganic chemistry, in research and industry. It is used in quality
control, dynamic measurement, and monitoring applications such as the long-
term unattended measurement of CO2 concentrations in greenhouses and
growth chambers by infrared gas analyzers.
FTIR spectrometer
http://www.chemicool.com/definition/fourier_transform_infrared_spectrometer_ftir.html
ADVANTAGES / DISADVANTAGES
UV - VIS
ADVANTAGES DISADVANTAGES
A light source shutter controls the amount of light However, no single lamp emits all the light
from a specialized lamp that passes through the wavelengths necessary for analysis. For
sample. The shutter is the only moving component example, a deuterium lamp emits wavelengths
of a UV-vis spectrometer. The advantage of this from 180 nm to 370 nm, and a tungsten lamp
system lies in the simplistic design of the emits wavelengths from 315 nm to 900 nm.
instrument. Changing the lamp is a time-consuming process.
Sample analysis using UV-vis is a very quick UV-vis spectrometers require frequent
process compared to other methods of sample calibrations to retain the accuracy and precision
detection, such as HPLC. This rapid analysis is of the instrument. Choosing what type of
achieved only through proper calibration. material to use as the calibrator requires the
knowledge of the type of sample being analyzed.
The UV-vis technique is non-destructive to the However, stray light can be a problem for UV-
sample and has a high sensitivity for detecting Vis spectrometers. This can be caused by the
organic compounds. user trying to detect the sample using too wide
of a wavelength range or by poor instrument
design.
ADVANTAGES / DISADVANTAGES
FT - IR
ADVANTAGES DISADVANTAGES
The multiplex advantage arises because all of the FTIR instruments do not measure spectra; they
spectral elements are measured simultaneously. measure interferograms which are difficult to
Thus, a spectrum can be obtained very quickly. interpret without performing a Fourier
transsform to produce a spectrum.
The throughput advantage arises because unlike In systems which are source noise limited, the
the dispersive spectrometers, FTIR disadvantage arises from the fact that if noise
spectrometers have no slits which attenuate the occurs in one part of the radiation from the
infrared light. infrared source, it will spread throughout the
spectrum.
The frequency scale of the spectrum is known FTIR instruments have a single beam, and for
very accurately. highly sensitive work experiments can take a
long time and changes in infrared absorbing gas
concentrations can severely affect the results.
CONCLUSIONS
• The use of spectrophotometry as it results from the presented

materials proves to be of real help in regards to analyzing the
properties of different materials from the point of view of
their chemical properties.
• The large variety of methods used and equipment typology

shows the technological development in this field of interest
relative to its possibilities in analyzing materials in different
domains i.e. medical, industrial, etc.
THANK YOU FOR YOUR KIND
ATTENTION !

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EUROPEAN UNION Structural Funds

GOVERNMENT OF ROMANIA GOVERNMENT OF THE REPUBLIC 2007 - 2013

SPECTROPHOTOMETRY. PRINCIPLE AND APPLICATIONS

As.dr.ing. ADRIAN EUGEN CIOABLA

In the frame of the project

Sustainable development of an research center in Banat region and

Ultraviolet–visible spectroscopy or ultraviolet-visible spectrophotometry

UV/Vis spectroscopy is routinely used in analytical chemistry for the

Infrared spectroscopy (IR spectroscopy) is the spectroscopy that deals with

Single beam spectrophotometer http://en.wikipedia.org/wiki/Spectrophotometry

Double beam spectrometer

Infrared spectroscopy is a simple and reliable technique widely used in both

• The use of spectrophotometry as it results from the presented

• The large variety of methods used and equipment typology

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