Spectrophotometer Mls 3rd

SPECTROPHOTOMET
ER
.
DEFINITION
Aspectrophotometer is an instrument that measures the
amount of light absorbed by a sample.
Spectrophotometer techniques are used to measure the
concentration of solutes in solution by measuring the amount
of the light that is absorbed by the solution in a cuvette placed
in the spectrophotometer.
PRINCIPLE OF
SPECTROPHOTOMETER
BEER – LAMBERT LAW
It states that the absorbance of light by a material in a solution is

directly proportional to its concentration in that solution.
A =c
Where,
A – absorbance
c – concentration
COMPONENTS OF
SPECTROPHOTOMETER
 The spectrometer consists of the following parts :
1.Light source 3. Monochromator 5. Detector
2.Collimator 4. wavelength selector

COMPONENTS OF
SPECTROPHOTOMETER
The spectrometer consists of the following parts :
1.Light source : it produce a desired range of

wavelength of light.
The function of the light source is to provide a
sufficient of light which is suitable for marking a
measurement. The light source typically yields a high
output of polychromatic light over a wide range of the
spectrum.
2. Collimator : transmits a straight beam of light.
COMPONENTS OF
SPECTROPHOTOMETER
3. Prism:- Prism is used to isolate different
wavelength .If a parallel beam of radiation falls on
a prism , the radiation of two different
wavelength will be bent through different angles.
4. Monochromator : split the light into its component wavelength.
Accepts polychromatic input light from a lamp a n d outputs

monochromatic light.
5. Wavelength selector : transmits only the desired wavelength.

COMPONENTS OF
SPECTROPHOTOMETER
6. CUVETTE:-
A cuvette is a kind of cell (usually a small square tube) sealed at one end,
made of Plastic, glass or optical grade quartz and designed to hold
samples for spectroscopic experiments. Cuvette should be as clear as
possible, without impurities that might affect a spectroscopic reading.
Like a test-tube, a cuvette may be open to the atmosphere on top or
have a glass or Teflon cap to seal it shut.
7. Detector :- Any photosensitive device can be used as a detector of

radiant energy .The photocell a n d phototube are the simplest
photodetectors, producing current proportional to the intensity of the
light striking Them .
SPECTROPHOTOMETER CAN BE
CLASSIFIED INTO TWO DIFFERENT TYPES :
SINGLE BEAM SPECTROMETER :
To measure the intensity of the incident light the sample
must be removed so that the reference can be placed each
time. This type of spectrometer is usually less expensive
and less complicated.
DOUBLE BEAM SPECTOMETER:
In this type, before it reaches the sample, the light source is

split into two separate beams. From these one passes
through the sample and second one is used for reference.
This gives an advantage because the reference reading and
sample reading can take place at the same time.
BASED ON THE WAVELENGTH OF LIGHT
USED IT CAN BE CLASSIFIED INTO :
VISIBLE SPECTROMETER
Uses visible range (400 – 700nm) of electromagnetic radiation spectrum.
Plastic and glass cuvettes can be used for visible light spectroscopy.
UV SPECTOMETER
Uses light over the UV range (180 - 400 nm).
UV spectroscopy is used for fluids, and even solids. Cuvettes, only made of
quartz, are used for placing the samples.
IR SPECTROPHOTOMETER
Uses light over infra red range (700 -15000) of electromagnetic radiation
spectra.
APPLICATIONS
1)Measure the concentration of the solution
A spectrophotometer optically determines the
absorbance or transmission of characteristic
wavelengths of radiant energy (light) by a chemical
species in solution. Each molecule absorbs light at
certain wavelengths in a unique spectral pattern.
APPLICATIONS
2) Identify organic compounds by determining the absorption
maximum.
Spectrophotometers are used to identify organic compounds by
determining the absorption maxima (which for most compounds
and groups of compounds have very distinct fingerprints (that's
what the absorption curves and peaks are called).
3) Used for color determination within the spectral range
If one is working in the range of 380 to 700 nm, the spectrophotometers
can also be used for color determination within this spectral range.
APPLICATIONS
 Detection of impurities:- UV absorption spectroscopy is one of the
best methods for determination of impurities in organic molecules.
Additional peaks can be observed due to impurities in the sample.
 Molecular weight determination:- molecular weight of compounds

can be measured spectrophotometrically by preparing the suitable
derivatives of these compounds. For e.g. if we want to determine
the molecular weight of amine then it is converted into amine
picrate.
.
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Spectrophotometer Mls 3rd

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SPECTROPHOTOMET

It states that the absorbance of light by a material in a solution is

 The spectrometer consists of the following parts :

1.Light source 3. Monochromator 5. Detector

2.Collimator 4. wavelength selector

1.Light source : it produce a desired range of

Accepts polychromatic input light from a lamp a n d outputs

5. Wavelength selector : transmits only the desired wavelength.

7. Detector :- Any photosensitive device can be used as a detector of

In this type, before it reaches the sample, the light source is

 Molecular weight determination:- molecular weight of compounds

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