SPECTROS

SPECTROSCOPY
 The study of the interaction between matter and electromagnetic radiation

 It is the term used to refer to the measurement of radiation intensity as a function of
wavelength.
 Often use to describe experimental spectroscopic methods.
 Four different spectroscopic techniques are commonly used and this include
1. Ultraviolet-visible spectroscopy
2. Infrared spectroscopy
3. Nuclear magnetic resonance spectroscopy (NMR)
4. Mass spectrometry.
The first three are known as absorption spectroscopy
 Spectral measurement devices are referred to as spectrometers, spectrophotometers,

spectrographs or spectral analyzers.
Spectrophotometers
 A spectrophotometer is an instrument that measures the amount of light absorbed by a

sample.
 Spectrophotometer techniques are mostly 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.
 Scientist Arnold J. Beckman and his colleagues at the National Technologies Laboratory
(NTL) invented the Beckman DU spectrophotometer in 1940.
Principle of Spectrophotometer
The spectrophotometer technique is to measure light intensity as a function of wavelength. It

does this by diffracting the light beam into a spectrum of wavelengths, detecting the intensities
with a charge-coupled device, and displaying the results as a graph on the detector and then on
the display device.
1. A sample solution is placed inside the spectrophotometer.

2. A light source shines light toward the sample.
3. A device called a monochromator splits the light into each color, or rather, individual
wavelengths (just like a raindrop makes a rainbow). An adjustable slit allows only one
specific wavelength of light through to the sample solution.
4. The wavelength of light hits the sample, which is held in a little container called a
cuvette. We need to be careful when handling cuvettes; even a slight fingerprint can
interfere with the results.
5. Whatever light passes through the sample is read and displayed on the output screen.
Instrumentation of Spectrophotometer
The essential components of spectrophotometer instrumentation include:
1. A table and cheap radiant energy source
 Materials which can be excited to high energy states by a high voltage electric discharge
(or) by electrical heating serve as excellent radiant energy sources.
2. A monochromator, to break the polychromatic radiation into component wavelength

(or) bands of wavelengths.
 A monochromator resolves polychromatic radiation into its individual wavelengths and

isolates these wavelengths into very narrow bands. It could be prism (or) grating.
PRISMS:
 A prism disperses polychromatic light from the source into its constituent wavelengths by
virtue of its ability to reflect different wavelengths to a different extent
 Two types of Prisms are usually employed in commercial instruments. Namely,
600 cornu quartz prism and 300 Littrow Prism.
GRATINGS:
 Gratings are often used in the monochromators of spectrophotometers operating

ultraviolet, visible and infrared regions.
3. Transport vessels (cuvettes), to hold the sample
 Samples to be studied in the ultraviolet (or) visible region are usually glasses (or)
solutions and are put in cells known as “CUVETTES”.
 Cuvettes meant for the visible region are made up of either ordinary glass (or) sometimes
Quartz.
4. A Photosensitive detector and an associated readout system

 Most detectors depend on the photoelectric effect. The current is then proportional to the
light intensity and therefore a measure of it.
 Radiation detectors generate electronic signals which are proportional to the transmitter
light.
 These signals need to be translated into a form that is easy to interpret.
 This is accomplished by using amplifiers, Ammeters, Potentiometers and Potentiometric
recorders.
Measuring Absorbance
The amount of light that makes it through the substance is displayed on the output screen and is
called absorbance. In order for this number to mean anything, we need a standard curve. A
standard curve is determined by recording the absorbance of known concentrations of a
material.
Applications of Spectrophotometer
Some of the major applications of spectrophotometers include the following:
 Detection of concentration of substances

 Detection of impurities
 Structure elucidation of organic compounds
 Monitoring dissolved oxygen content in freshwater and marine ecosystems
 Characterization of proteins
 Detection of functional groups
 Respiratory gas analysis in hospitals
 Molecular weight determination of compounds
 The visible and UV spectrophotometer may be used to identify classes of compounds in
both the pure state and in biological preparations.

SPECTROS

Uploaded by

Copyright:

Available Formats

SPECTROS

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

SPECTROS

Uploaded by

Copyright:

Available Formats

SPECTROSCOPY

 The study of the interaction between matter and electromagnetic radiation

The first three are known as absorption spectroscopy

 Spectral measurement devices are referred to as spectrometers, spectrophotometers,

 A spectrophotometer is an instrument that measures the amount of light absorbed by a

The spectrophotometer technique is to measure light intensity as a function of wavelength. It

1. A sample solution is placed inside the spectrophotometer.

The essential components of spectrophotometer instrumentation include:

1. A table and cheap radiant energy source

2. A monochromator, to break the polychromatic radiation into component wavelength

 A monochromator resolves polychromatic radiation into its individual wavelengths and

 Gratings are often used in the monochromators of spectrophotometers operating

3. Transport vessels (cuvettes), to hold the sample

4. A Photosensitive detector and an associated readout system

Some of the major applications of spectrophotometers include the following:

 Detection of concentration of substances

You might also like