# Spectophotometery
Uploaded by
Sibgha S# Spectophotometery
Uploaded by
Sibgha SSPECTOPHOTOMETERY
SPECTOPHOTOMETERY
A method in which the absorption or transmission
properties of a material is quantitatively
measured as a function of wavelength.
The basic principle behind this method is that :
“Each compound absorbs or transmit
light over a certain range of wavelength”
SPECTOPHOTOMETER
It is an instrument that measures the amount
of photons absorbed by a sample after it is
passed through its solution.
With the spectrophotometer , the amount of
known chemical substance can be determined
by measuring the absorbance.
It is divided into two types
1. Single beam spectrometer
2. Double beam spectrometer
SINGLE BEAM
SPECTROMETER
To measure the
intensity of the incident
light the sample must
be removed so the
reference can be
placed each time.
This type of
spectrometer is usually
less expensive and
less complicated.
DOUBLE BEAM
SPECTROMETER
In this type ,before it
reaches the sample the
light source is split into two
separate beams.
From these one passes
passes through the sample
and the second one is used
for reference.
This gives an advantage
because the reference
reading and sample reading
can take place at the same
time.
PRINCIPLE OF
SPECTROPHOTOMETRY
Spectrophotometry is a method to measure how
much a chemical substance absorbs light by
measuring the intensity of light as a been of light
passes through sample solution.
The basic principle is that each compound
absorbs or transmit light over a certain ranges of
wavelength.
APPLICATIONS OF
SPETROPHOTOMETRY
Following are the applications are of
spectrophotometry
1. Measurement of concentration
2. Detection of impurities
3. Chemical kinetics
4. Detection of functional group
5. Molecular weight determination
MEASUREMENT OF
SAMPLE
Prepare sample
Make series of standard solutions of known
concentrations
Set spectrophotometer to the λ of maximum light
absorption
Measure the absorption of the unknown , and from the
standard plot, read the related concentration.
DETECTION OF
IMPURITIES
UV absorption spectroscopy is one of the best
method for determination of impurities in organic
molecule.
Additional peaks can be observed due to
impurities in the sample and it can be compared
with that of standard raw material.
CHEMICAL KINETICS
Kinetics of reaction can also be studied
using UV spectroscopy.
The UV radiation is passed through the
reaction cell and the absorbance changes
can be observed.
DETECTION OF
FUNCTIONAL GROUP
Absence of a band at particular wavelength
regarded as an evidence for absence of a
particular group.
MOLECULAR WEIGHT
DETERMINATION
Molecular weight of compound can be
measured specrtophotometrically by
preparing the suitable derivatives of these
compounds.
Example :
if we want to determine the
molecular weight of amine then it is converted
into amine picrate.
