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Introduction To Spectros

Spectroscopy is the study of matter's properties through its interaction with electromagnetic spectrum frequencies, providing insights into molecular states and structures. UV-Vis spectroscopy specifically measures light absorbance in the ultraviolet and visible ranges, allowing for the quantification of analytes based on their absorption characteristics. While UV-Vis spectrometers offer high accuracy and ease of use, they require careful setup to avoid interference from external light and noise.

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Eman Mohamed
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15 views20 pages

Introduction To Spectros

Spectroscopy is the study of matter's properties through its interaction with electromagnetic spectrum frequencies, providing insights into molecular states and structures. UV-Vis spectroscopy specifically measures light absorbance in the ultraviolet and visible ranges, allowing for the quantification of analytes based on their absorption characteristics. While UV-Vis spectrometers offer high accuracy and ease of use, they require careful setup to avoid interference from external light and noise.

Uploaded by

Eman Mohamed
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PPTX, PDF, TXT or read online on Scribd
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Introduction to Spectroscopy

What is spectroscopy?
 Studying the properties of matter through its interaction with different
frequency components of the electromagnetic spectrum.
With light, you aren’t looking directly at the molecule—the matter—but
its “ghost.” You observe the light’s interaction with different degrees of
freedom of the molecule.
Each type of spectroscopy—different light frequency—gives adifferent
picture → the spectrum.
Spectroscopy is a general methodology that can be adapted in many
ways to extract the information you need (energies of electronic,
vibrational, rotational states, structure and symmetry of molecules,
dynamic information).
What does a spectrum
measure?
• Interaction of light with a sample can influence the sample and/or the
light. Method involves:
• (1) excitation
• (2) detection.
In most spectroscopies, we
characterize how a sample
modifies light entering it.
• 1- Absorption: Change in intensity I of incident light
• Sample attenuates light → transmission T= I/I0

We measure the absorption of light at different


frequency or wavelength.
ω/λ/ν (characteristic frequency/wavelength
of light entering sample)
• 2- Emission: Excitation induces emission of light from the sample
(usually of different frequency).

Includes:
I. Fluorescence (emission from excited electronic singlet states)
II.Phosphorescence (emission from excited electronic triplet states)
III. Raman Scattering (light scattering involving vibrational transition)
3- Optical Rotation:
Change of phase of light incident on sample (rotation of polarization)
What are the axes?
• 1.X-axis: Characterizes the input light in terms of frequency-
wavelength-energy.
• 2- y-axis: Absorption
• This comes from assuming that the fraction of light absorbed as you
propagate though the sample is proportional to the distance
traversed: dI / dx = −α
How do your measure
absorption spectra?
• Measure the change of intensity of light at different frequencies as it
passes through a sample.Two types of spectrometers:
• 1) Dispersive
• 2) Fourier transform
• Dispersive spectrometer: Separate different frequency components
What is a UV-Vis Spectrophotometer?
Ultraviolet-visible (UV-Vis) spectrophotometry is a
technique used to measure light absorbance across the
ultraviolet and visible ranges of the electromagnetic
spectrum.
When incident light strikes matter it can either be
absorbed, reflected, or transmitted.
The absorbance of radiation in the UV-Vis range causes
atomic excitation, which refers to the transition of
molecules from a low-energy ground state to an excited
state.
A UV-Vis spectrophotometer can use this principle to
quantify the analytes in a sample based on their
absorption characteristics.
What is analyzed when
ultraviolet-visible
spectroscopy?

UV-Vis spectroscopy is an analytical technique that


measures the amount of discrete wavelengths of UV
or visible light that are absorbed by or transmitted
through a sample in comparison to a reference or
blank sample.
How does a UV-Vis
spectrophotometer work?
Instrumentation of UV
Spectroscopy
• he basic components of a spectrometer include: light source (UV and
visible), monochromator (wavelength selector), sample stage, and
detector.
• 1- Light source (UV and visible):
• •As a light-based technique, a steady source able to emit light across
a wide range of wavelengths is essential.
• •A single xenon lamp is commonly used as a high intensity light
source for both UV and visible ranges.
• •Xenon lamps are, however, associated with higher costs and are less
stable in comparison to tungsten and halogen lamps.
• 2- Monochromator (wavelength selector):
• A monochromator separates light into a narrow band of
wavelengths. It is most often based on diffraction gratings
that can be rotated to choose incoming and reflected angles
to select the desired wavelength of light.
• 3- Sample analysis : Whichever wavelength selector is used
in the spectrophotometer, the light then passes through a
sample. For all analyses, measuring a reference sample,
often referred to as the "blank sample", such as a cuvette
filled with a similar solvent used to prepare the sample, is
imperative. If an aqueous buffered solution containing the
sample is used for measurements, then the aqueous
buffered solution without the substance of interest is used as
the reference.
• 4- Detector:
• After the light has passed through the sample, a detector is
used to convert the light into a readable electronic signal.
Generally, detectors are based on photoelectric coatings or
semiconductors.
Advantages of UV-VIS
Spectrometers
• The biggest advantage for chemists and astronomers who use UV-VIS
spectrometers is the accuracy of the device. Even small UV-VIS
spectrometers can give extremely accurate readings, which is crucial
when you are preparing chemical solutions or recording the
movement of celestial bodies.
• UV-VIS spectrometers are easy to use. Most UV-VIS spectrometers
used in astronomy attach to telescopes. Most of the ones used in
chemistry are comparable in size to electron microscopes and require
the same basic skills to use. Because they are simple to operate, there
is little chance of a UV-VIS spectrometer being used improperly.
Disadvantages of UV-VIS
Spectrometers
• The main disadvantage of using a UV-VIS spectrometer is the time it
takes to prepare to use one. With UV-VIS spectrometers, setup is key.
You must clear the area of any outside light, electronic noise, or other
outside contaminants that could interfere with the spectrometer's
reading.
• If the space has been properly prepared ahead of time, UV-VIS
spectrometers are simple to use and give accurate results. However, if
the space has not been properly prepared, even a small bit of outside
light or vibration from a small electronic device could interfere with
the results you are hoping to achieve in using a UV-VIS spectrometer.
What are the applications of
ultraviolet spectroscopy?
• Ultraviolet-visible (UV-Vis) spectroscopy is a widely used technique in many
areas of science ranging from :
• 1.Quality control.
• 2.Cosmetic industry.
• 3.Petrochemistry.
• 4.Pharmaceutical research.
• 5.Optical components.
• 6.Food and agriculture.
• 7.Life sciences.
• 8.Traditional chemistry

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