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    UV Visible Spectros

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    M Hmza S Malik
    UV spectroscopy involves using ultraviolet light between 200-400 nm to excite electrons in molecules from their ground state to higher energy states. UV-visible spectrometers measure the absorbance of UV or visible light by a sample at single or multiple wavelengths. The spectrometer contains a light source, diffraction grating to split light by wavelength, and a detector. The Beer-Lambert law states that absorbance is proportional to concentration and path length, and is used to determine sample concentrations from their absorbance measurements. UV-Vis spectroscopy has applications in reaction kinetics, quantification of biomolecules, and environmental analysis.

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    UV Visible Spectros

    Uploaded by

    M Hmza S Malik
    84 views2 pages
    UV spectroscopy involves using ultraviolet light between 200-400 nm to excite electrons in molecules from their ground state to higher energy states. UV-visible spectrometers measure the absorbance of UV or visible light by a sample at single or multiple wavelengths. The spectrometer contains a light source, diffraction grating to split light by wavelength, and a detector. The Beer-Lambert law states that absorbance is proportional to concentration and path length, and is used to determine sample concentrations from their absorbance measurements. UV-Vis spectroscopy has applications in reaction kinetics, quantification of biomolecules, and environmental analysis.

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    UV Visible Spectroscopy

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    UV spectroscopy involves using ultraviolet light between 200-400 nm to excite electrons in molecules from their ground state to higher energy states. UV-visible spectrometers measure the absorbance of UV or visible light by a sample at single or multiple wavelengths. The spectrometer contains a light source, diffraction grating to split light by wavelength, and a detector. The Beer-Lambert law states that absorbance is proportional to concentration and path length, and is used to determine sample concentrations from their absorbance measurements. UV-Vis spectroscopy has applications in reaction kinetics, quantification of biomolecules, and environmental analysis.

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    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    84 views2 pages

    UV Visible Spectros

    Uploaded by

    M Hmza S Malik
    UV spectroscopy involves using ultraviolet light between 200-400 nm to excite electrons in molecules from their ground state to higher energy states. UV-visible spectrometers measure the absorbance of UV or visible light by a sample at single or multiple wavelengths. The spectrometer contains a light source, diffraction grating to split light by wavelength, and a detector. The Beer-Lambert law states that absorbance is proportional to concentration and path length, and is used to determine sample concentrations from their absorbance measurements. UV-Vis spectroscopy has applications in reaction kinetics, quantification of biomolecules, and environmental analysis.

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    Introduction to Ultraviolet -Visible Spectroscopy (UV)

    Principle
    UV spectroscopy is type of absorption spectroscopy in which light of ultra-violet region (200-400 nm.) is
    absorbed by the molecule. Absorption of the ultra-violet radiations results in the excitation of the electrons
    from the ground state to higher energy state. The energy of the ultra-violet radiation that are absorbed is
    equal to the energy difference between the ground state and higher energy states (ΔE = hf).

    UV-Visible Spectrometer
    UV-visible spectrometers can be used to measure the absorbance of ultra violet or visible light by a
    sample, either at a single wavelength or perform a scan over a range in the spectrum. The UV region
    ranges from 190 to 400 nm and the visible region from 400 to 800 nm. The technique can be used both
    quantitatively and qualitatively. A schematic diagram of a UV-visible spectrometer is shown.

    The light source (a combination of tungsten/halogen and deuterium lamps) provides the visible and near
    ultraviolet radiation covering the 200 – 800 nm. The output from the light source is focused onto the
    diffraction grating which splits the incoming light into its component colours of different wavelengths,
    like a prism (shown below) but more efficiently.
    It is possible to predict which wavelengths are likely
    to be absorbed by a coloured substance. When white
    light passes through or is reflected by a coloured
    substance, a characteristic portion of the mixed
    wavelengths is absorbed. The remaining light will
    then assume the complementary colour to the
    wavelength(s) absorbed.

    For liquids the sample is held in an optically flat,


    transparent container called a cell or cuvette. The reference cell or cuvette contains the solvent in which
    the sample is dissolved and this is commonly referred to as the blank.
    For each wavelength the intensity of light passing through both a reference cell (Io) and the sample cell (I)
    is measured. If I is less than Io, then the sample has absorbed some of the light.
    The absorbance (A) of the sample is related to I and Io according to the following equation:

    A=I/Io………………….. (1)
    The detector converts the incoming light into a current, the higher the current the greater the intensity. The
    chart recorder usually plots the absorbance against wavelength (nm) in the UV and visible section of the
    electromagnetic spectrum. (Note: absorbance does not have any units).

    The Beer-Lambert Law

    According to the Beer-Lambert Law the absorbance is proportional to the concentration of the
    substance in solution and as a result UV-visible spectroscopy can also be used to measure the
    concentration of a sample.
    The Beer-Lambert Law can be expressed in the form of the following equation:
    A = cl
    Where
    A = absorbance
    l = optical path length, i.e. dimension of the cell or cuvette (cm)
    c = concentration of solution (mol dm-3)

    Applications

     UV/Vis can be applied to determine the kinetics or rate constant of a chemical reaction
     The concentration of the sample in the sample can be determined
     In the biochemical and genetic fields UV-visible spectroscopy is used in the quantification of DNA.
     In environmental and agricultural fields the quantification of organic materials and heavy metals in fresh
    water can be carried out using UV-visible spectroscopy.

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