Uv Vis

UV-VIS
SPECTROSCOPY
B T P 47 5 3 T E C H I N N O F O R P H A R M A E N G I N E E R I N G
PREPARED BY;
NUR SHAHIRA FIZUNI FARHAH BINTI ROSLAN TD14020
VIVIANA ABDULLAH TD14025
INTRODUCTION
Spectroscopy
 The study of absorbance and/or emission of electromagnetic radiation by
a particular molecule and the correlation with details of molecular
structure
Electromagnetic radiation
 EMR consist of discrete packages of energy which called as
photons
 Frequency, v is the number of times electrical field radiation
oscillates in once second (unit of frequency is Hertz, Hz)
 Wavelength, λ is the distance between two nearest parts of the
wave in the same phase
INTRODUCTION
Ultraviolet (UV) and Visible (VIS) Spectroscopy
 Measurement of electromagnetic radiation after it absorb by a sample or after
reflection from a sample surface
 UV spectrum range from 200nm to 400 nm
 VIS spectrum range from 400 nm to 800 nm
Basic components of UV-Vis Spectroscopy
1. Radiation Source
 Hydrogen discharge lamp that consist of two electrode contain in
deuterium filled silica envelop
2. Monochrometer
 Consists of two slits that separated by a prism
3. Beam Splitter
 Bend the monochromatic light and produce two equal parallel beam of light
4. Sample Compartments
 Consists of transparent reference cell and sample cell loadout
5. Detector
 Device that converted impact of photons into electrical current that can be
monitor by computer
Basic components of UV-Vis Spectroscopy
WORKING PRINCIPLE
LIGHT SOURCE
• Tungsten filament lamps

and Hydrogen-Deuterium
lamps are most widely used
as they cover the whole UV
region.
MONOCHROMATOR
• generally composed of
prisms and slits.
• Most are double beam
spectrophotometers
• Radiation emitted from
primary source is dispersed
by the rotating prisms.
BEAM SPLITTER
• Separate bend and send it

to sample and reference
cells.
SAMPLE AND REFERENCE
CELLS
• One of the divided beams is
passed through the sample
solution, second beam
passes through the
reference solution.
• Sample and reference are
contained in cells made up
of silica or quartz.
• Glass cant be used as it
absorbs the UV region
DETECTORS
• Two photocells are used as
the detector in UV
spectroscopy.
AMPLIFIER
• Amplify the signals many

times to get clear and
recordable signals.
RECORDING DEVICE
• Data generated and

produces the spectrum of
the desired compound.
HISTORY OF UV-VIS
ADVANCEMENT OF SPECTROSCOPY
UV-VIS SPECTROSCOPY
• In 1940s, the American government spurred interest in measuring vitamin content in
soldiers’ rations using UV-Vis spectroscopy, this research culminated the commercial launch
of UV-Vis spectroscopy
• Launch of first UV-Vis spectroscopy by the Beckman Instruments
• In 1940s, Becman introduces DU UV-Vis, which has higher resolution and lower stray light in
the UV region
• Starting 1950s, mass production reduces the cost of UV-Vis. A new implemented of
photodiode arrays collect all wavelengths simultaneously, reducing time required to scan
spectrum to seconds
• 1990,s External software that provides PC control, onscreen spectra display and spectra
reprocessing and storage. Fiber optics reduce instrument size.
• 2000s, Ability of UV-Vis to measure micro volume liquid samples (<1μL) in biotechnology and
pharmaceutical applications
• 2010, Thermo Scientific introduced UV-Vis of its new application that focused beam geometry
tailors the instruments optical system to specific application for microcells, solid sampling
and fiber optics
TYPES OF UV-VIS SPECTROSCOPY
1. Single Beam Spectroscopy
• Light from radiation source pass through monochromator enters sample cell
containing the sample solution
• The absorbance readings of both standard and unknown solutions are recorded
after adjusting the instrument to 100% transmittance with blank solution each
time the wavelength is changed
TYPES OF UV-VIS SPECTROSCOPY
2. Double Beam Spectroscopy
• Allows direct measurement ratio of intensities of sample and reference beams
respectively
• Similar to single beam except it contains beam slitter or chopper
APPLICATION IN PHARMACEUTICAL INDUSTRIES
1. Detection of impurities
• UV/Vis spectroscopy can be used in the manufacturing process to identify
contaminants within a substance
• By comparing the spectrum obtained with that of standard raw material, additional
peaks can be observed if impurities are present in the sample
2. Quantitative analytical of pharmaceutical substances
• It can be used for the routine analysis of drugs in pure and pharmaceutical
formulations
• It is carried out by preparing a suitable solution of the drug in a solvent and
measuring its absorbance at specific wavelength
3. Qualitative Analysis
• Identifying an unknown compound can be determined through the simple process
of UV/Vis spectroscopy
• The process involves comparing its absorption spectrum with the spectra of
known compounds
• But it is bit applicable for characterizing those types of compounds which do not
absorb UV radiation
Qualitative
Analysis
Organic Inorganic
compounds compounds
• Determination of optical densities of bacterial culture

• Determination of the amount of drug reaching each body part
• Measure the chemical kinetics of a reaction
• Molecular weight determination
• Dissociation constants of acids and bases
DATA ANALYSIS
• Calibration is the process of
obtaining the response of the
system (absorbance) to a known
input (concentration of the
solution).
• The curve is used to determine
the concentration of a substance
by measuring its absorbance.

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UV-VIS

• Tungsten filament lamps

• Separate bend and send it

• Amplify the signals many

• Data generated and

• Determination of optical densities of bacterial culture

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