Govt.

Post Graduate College Haripur

SPECTROPHOTOMETER

Presented by:
M. ZAIN ALI KHAN

Presented To : SIR KAMRAN MEHDI

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 Introduction
SYNOPSIS
 MEANING OF SPECTROPHOTOMETER

 INTRODUCTION

 PRINCIPLE

 TYPES OF INSTRUMENT

 TYPES OF SPECTROPHOTOMETER

 UV-VIS SPECTOPHOTOMETER

 INFRA RED SPECTOPHOTOMETER

 APPLICATION
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 Introduction
MEANING OF SPECTROPHOTOMETRY

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 Introduction
INTRODUCTION: SPECTROPHOTOMETER

 An instrument that measures the amount of light that passes

through (is transmitted through) a sample.
 It is a photometer (a device for measuring light intensity) that can
measure intensity as a function of the color (or more specifically
the wavelength).
 This means it use light in the visible and adjacent (near ultraviolet
and near infrared) range

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 Introduction
PRINCIPLE
 When a beam of monochromatic light passed through a solution it
may transmitted as a such or some of may be absorb.
 Proportional the transmitted light can be represented by intensity
of the incident radiation.
T=I/I₀
Absorbance(A) of light through a solution in inversely proportional
to log₁₀ of %T
A=Log(₁∕T)
=Log(I/I₀)
Where I=intensity of transmitted light
I₀=intensity of incident light

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 Introduction
PRINCIPLE
 The quantitative determination of compounds by spectrometric
technique is based on two law.
1 Lamberts Law
2 Beers Law
 Lamberts law- its state that light absorbed by solution is directly
proportional to length of the light through the solution.
hence
A=log(Io/I)= Є.C
where A= absorbance
Є=molar absorptivity coefficient
l= path length of the sample (usually 1cm)

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PRINCIPLE
 Beers law -its state that the amount of light absorbed is directly proportional to
concentration of absorbing solute in the solution.
Thus
A=log(Io/I)= Є.C
where C=concentration of solution moles liter-1
combining equation
A=log(Io/I)= Є.C.l
it a standard cuvette with light path of 1.cm is used.
A=log(Io/I)= Є.C
Limitation of Beers Lamberts Law:
 when different forms of the absorbing molecules are in equilibrium as in keto –
enol tautomers.
 when fluorescent compounds are present.
 when solute and solvent forms complex though some sort of association .
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 Introduction
TYPES OF INSTRUMENT
 Single beam-A single beam spectrophotometer measures the
light intensity of the beam and after a test sample is inserted.

 Double beam - A double beam spectrophotometer compares

the light intensity b/w two path.one path containing a
reference sample & the other is test sample.

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TYPES OF INSTRUMENT

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 Introduction
TYPES OF SPECTROPHOTOMETER

 Ultraviolet (UV)-Vis Spectrophotometers Uses ultraviolet

light of wave lengths from 200 nm to 350 nm
 Infra Red (Ir)- covers the range of the electromagnetic
spectrum between 0.78 and 1000 m, m. In the context of
infra red spectroscopy wavelength is measured in "wave
numbers", which have the units cm-1. The most useful I.R.
region lies between 4000 - 670cm-1.

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 Introduction
TYPES OF SPECTROPHOTOMETER

 UV-visible spectroscopy involve the spectroscopy of

photons in the UV-visible region. This means it use light in
the visible and adjacent (near UV and near Infrared)range.
 It uses light over the ultraviolet range (185- 400 nm) and
visible range(400-700 nm) of electromagnetic radiation
spectrum.

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INSTRUMENTATION

Spectrophotometer have the following basic components.

Source

Filter and monochromators

Cell / sample holder

Detector

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INSTRUMENTATION
 Source:- A continuous source of radiant energy covering
the region of spectrum in which the instrument is designed
to work. UV- Hydrogen Deuterium lamp Visible- Tungsten
lamp.
 Filter and monochromators:- A light filter is a device that
allow light of the required wavelength to pass but absorbs
light of the other wavelength wholly and partially.
 Filter-:
Filter- Filter are of two types
 Absorption filter
 Interference filter

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INSTRUMENTATION
 Monochromators: The monochromator is to dispersing the
radiation according to the wavelength.
 dispersing element (a prism or grating)
 Slits-:
Slits- there are two types of slit
 Entrance slit
 Exit slit
 Cell / sample holder :- The cell holding the sample (usually
a solution should be transparent to the wavelength region
being recorded.

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INSTRUMENTATION
 Detector-: In order to detect radiation , three types of
photosensitive device are-
 Photovoltaic cell
 Phototube
 Photomultiplier Photovoltaic cell Phototube
 Power supply:- The power supply serves as triple function-
 It decreases the line voltage to the instrument operating level with a
transformer.
• It convert A.C to D.C with a rectifier if direct is required by the
instrument .
• It smooth's out any ripple which may occur in the line voltages in
order to deliver a constant voltage to the source lamp and
instrument. 215
 Introduction
APPLICATION
 Qualitative analysis
 Quantitative analysis
 Study of cis trams isomerism
 Structure elucidation of organic compound
 Chemical kinetics-Kinetics of reaction can also be studied using UV
spectroscopy.
 Molecular weight determination-Molecular weights of compounds
can be measured spectrophotometrically by preparing the suitable
derivatives of these compounds.
 Extent of conjugation

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