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    Uv Visible Spectros

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    joshishravan3003
    UV-visible spectroscopy involves using light in the ultraviolet and visible regions to analyze molecules. It works by measuring how much light is absorbed by a sample at different wavelengths. There are two main types - single beam which passes light through a sample, and double beam which compares light passed through a sample versus a reference. It has many applications including detecting impurities, determining molecular structure, and quantifying concentration through the Beer-Lambert law. It is a simple, cost-effective technique commonly used in various fields like chemistry, medicine, environmental testing and forensics.

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    Uv Visible Spectros

    Uploaded by

    joshishravan3003
    120 views28 pages
    UV-visible spectroscopy involves using light in the ultraviolet and visible regions to analyze molecules. It works by measuring how much light is absorbed by a sample at different wavelengths. There are two main types - single beam which passes light through a sample, and double beam which compares light passed through a sample versus a reference. It has many applications including detecting impurities, determining molecular structure, and quantifying concentration through the Beer-Lambert law. It is a simple, cost-effective technique commonly used in various fields like chemistry, medicine, environmental testing and forensics.

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    study material pharmacy.

    Original Title

    UV VISIBLE SPECTROSCOPY

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    UV-visible spectroscopy involves using light in the ultraviolet and visible regions to analyze molecules. It works by measuring how much light is absorbed by a sample at different wavelengths. There are two main types - single beam which passes light through a sample, and double beam which compares light passed through a sample versus a reference. It has many applications including detecting impurities, determining molecular structure, and quantifying concentration through the Beer-Lambert law. It is a simple, cost-effective technique commonly used in various fields like chemistry, medicine, environmental testing and forensics.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    120 views28 pages

    Uv Visible Spectros

    Uploaded by

    joshishravan3003
    UV-visible spectroscopy involves using light in the ultraviolet and visible regions to analyze molecules. It works by measuring how much light is absorbed by a sample at different wavelengths. There are two main types - single beam which passes light through a sample, and double beam which compares light passed through a sample versus a reference. It has many applications including detecting impurities, determining molecular structure, and quantifying concentration through the Beer-Lambert law. It is a simple, cost-effective technique commonly used in various fields like chemistry, medicine, environmental testing and forensics.

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    © All Rights Reserved
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    Download as pptx, pdf, or txt
    of 28

    UV VISIBLE

    SPECTROSCOPY

    Presented by
    Mr. Rajkumar B. Patil
    M.Pharm 1st year
    Department of Pharmaceutics
    Ashokrao Mane College of Pharmacy
    Peth Vadgaon
    Definition
     Spectroscopy is the measurement and interpretation of electromagnetic

    radiation absorbed or emitted when the molecules or atoms or ions of a


    sample move from one energy state to another energy state.
     UV spectroscopy is a type of absorption spectroscopy in which light of the

    ultra-violet region (200-400 nm) is absorbed by the molecule which results


    in the excitation of the electrons from the ground state to a higher energy
    state.
     To Understand Spectroscopy We Must Understand Electromagnetic

    Radiation
     What is Electromagnetic Radiation?

     Is a form of energy that has both Wave and Particle Properties. For

    example: Ultraviolet, visible, infrared, microwave, radio wave.


    WAVE PROPERTIES

    EM radiation is conveniently modeled as waves consisting of perpendicularly


    oscillating electric and magnetic fields, as shown below

    Electric Field

    Magnetic Field

    Direction of
    propagation
     At 90° to the direction of propagation is an oscillation in the ELECTRIC

    FIELD.
     At 90° to the direction of propagation and 90° from the electric field

    oscillation (orthagonal) is the MAGNETIC FIELD oscillation.


    Wavelength (l)
    Electric Field

    Amplitude (A)

    Time or Distance

    We Use Symbols to Designate the Various Properties


    of Waves
    lis the wavelength of the waves
    v the frequency of the waves
    c is the speed of light
    Purpose of each Electromagnetic Radiation:
    Principle:
     Basically, spectroscopy is related to the interaction of light with matter.

     As light is absorbed by matter, the result is an increase in the energy content

    of the atoms or molecules.


     When ultraviolet radiations are absorbed, this results in the excitation of the

    electrons from the ground state towards a higher energy state.


     Molecules containing π-electrons or nonbonding electrons (n-electrons) can

    absorb energy in the form of ultraviolet light to excite these electrons to


    higher anti-bonding molecular orbitals.
     The more easily excited the electrons, the longer the wavelength of light they

    can absorb. There are four possible types of transitions (π–π*, n–π*, σ–σ*,
    and n–σ*), and they can be ordered as follows: σ–σ* > n–σ* > π–π* > n–π*
    Instrumentation:
    Instrumentation or Parts of UV Spectroscopy

    1. Light Source
     Tungsten filament lamps and Hydrogen-Deuterium lamps are the most

    widely used and suitable light sources as they cover the whole UV region.
     Tungsten filament lamps are rich in red radiations; more specifically they

    emit the radiations of 375 nm, while the intensity of Hydrogen-Deuterium


    lamps falls below 375 nm.
    2. Monochromator
     Monochromators generally are composed of prisms and slits.

     Most of the spectrophotometers are double beam spectrophotometers.

     The radiation emitted from the primary source is dispersed with the help of

    rotating prisms.
     The various wavelengths of the light source which are separated by the

    prism are then selected by the slits such the rotation of the prism results in a
    series of continuously increasing wavelengths to pass through the slits for
    recording purposes.
     The beam selected by the slit is monochromatic and further divided into

    two beams with the help of another prism.


    3. Sample and reference cells
     One of the two divided beams is passed through the sample solution and the

    second beam is passé through the reference solution.


     Both sample and reference solution is contained in the cells.

     These cells are made of either silica or quartz. Glass can’t be used for the

    cells as it also absorbs light in the UV region.


    4. Detector
     Generally, two photocells serve the purpose of the detector in UV

    spectroscopy.
     One of the photocells receives the beam from the sample cell and the second

    detector receives the beam from the reference.


     The intensity of the radiation from the reference cell is stronger than the

    beam of the sample cell. This results in the generation of pulsating or


    alternating currents in the photocells.
    5. Amplifier
     The alternating current generated in the photocells is transferred to the

    amplifier.
     The amplifier is coupled to a small servometer.

     Generally, the current generated in the photocells is of very low intensity,

    the main purpose of the amplifier is to amplify the signals many times so
    we can get clear and recordable signals.
    6. Recording devices
     Most of the time amplifier is coupled to a pen recorder which is connected

    to the computer.
     The computer stores all the data generated and produces the spectrum of the

    desired compound.
     Types of UV-Vis spectrometers

    There are two types of UV-vis spectrophotometers

    1. Single beam UV-Vis spectrometer

    2. Double beam UV-Vis spectrometer


    Single beam UV-Visible spectrophotometer
     Single beam uv-vis spectrophotometer has a single beam as the name

    indicates.
     The incident light coming from the source is passed through a

    monochromator then that incident monochromatic light moves through a


    slit.
     Then it passes through the sample solution. Where some of the incident

    light is absorbed by the sample while other is transmitted.


     That transmitted light is detected by the detector.

     The detected light is then amplified, recorded, and then displayed on a

    suitable readout device. Spectrum is plotted and the λ max is located.


    Double beam UV-Visible spectrophotometer
    The instrumentation of single and double beam spectrophotometers is almost
    the same.
    The basic difference from a single beam UV-Vis spectrophotometer is that the
    beam of incident light is passing simultaneously from the sample and the
    reference cells.
    The incident light splits and is directed towards both the reference and sample
    cuvette.
    The refracted or transmitted beam is detected by the detectors. A double beam
    UV-vis spectrophotometer needs two detectors that detect electron ratio to
    measure or calculate absorbance in a test sample. It also requires a stabilized
    voltage supply.
    Applications of UV Spectroscopy:

     Detection of Impurities

     It is one of the best methods for the determination of impurities in organic

    molecules.
     Additional peaks can be observed due to impurities in the sample and it can

    be compared with that of standard raw material.


     By also measuring the absorbance at a specific wavelength, the impurities

    can be detected.
     Structure elucidation of organic compounds

     UV absorption spectroscopy can be used for the quantitative


    determination of compounds that absorb UV radiation.
     UV absorption spectroscopy can characterize those types of compounds

    that absorb UV radiation thus used in the qualitative determination of


    compounds. Identification is done by comparing the absorption spectrum
    with the spectra of known compounds.
     This technique is used to detect the presence or absence of a functional

    group in the compound. The absence of a band at a particular wavelength is


    regarded as evidence for the absence of particular group.
     Kinetics of reaction can also be studied using UV spectroscopy. The UV

    radiation is passed through the reaction cell and the absorbance changes can
    be observed.
     Many drugs are either in the form of raw material or in the form of the

    formulation. They can be assayed by making a suitable solution of the drug


    in a solvent and measuring the absorbance at a specific wavelength.
     Molecular weights of compounds can be measured spectrophotometrically

    by preparing the suitable derivatives of these compounds.


     UV spectrophotometer may be used as a detector for HPLC.
     UV-visible spectroscopy is used to identify organic and inorganic

    species present in a solution.

     It is used to find the concentration of the unknown solution.

     For the determination of structure along with other data such as bands

    and intensities of functional groups uv-visible spectroscopy is used.

     It is also used in the study of chemical kinetics i.e. disappearance of one

    functional group and appearance of another functional group.

     UV-visible spectroscopy is used to study isomers, i.e. in geometric

    isomerism, the trans-species absorb a high wavelength with a large

    molar absorptivity ‘ε’ value than the cis-species.


     It helps in the detection or presence of conjugation.

     UV-vis spectroscopy is used in clinics and hospitals for drug analysis.

     It is used in petrochemical industries.

     UV-visible spectroscopy is used in water quality control labs.

     It is used in forensic labs.

     Another important application of UV-visible spectroscopy is in the field of

    chemical and biological plants.


     For qualitative analysis maximum absorption is used and for quantitative

    analysis, Lambert beer’s law is used.


    Conclusion
     UV-visible spectroscopy is a valid, simple and cost effective method for

    determining the concentration of absorbing species if applied to pure


    compounds, and used with the appropriate standard curve.
     A standard curve relating absorbance to concentration can be developed for

    any compound, and used to determine the concentration of samples


    containing the same compound.
     The analysis should be done at a wavelength with maximum absorption,

    and located in relatively flat region of the spectra so that absorbance will be
    high and constant in a narrow range around the chosen wavelength.
    References:
     http://www.indiastudychannel.com/resources/146681-Principle-working-

    and-applications-of-UV-spectroscopy.aspx
     https://www.slideshare.net/AlexaJacob1/uv-visible-spectroscopy-ppt

     https://www.slideshare.net/manishpharma/application-of-uv-spectroscopy

     https://medium.com/@ankur1857/principle-of-ultra-violet-uv-

    spectrophotometer-e6a1c435d258
     https://en.wikipedia.org/wiki/Ultraviolet%E2%80%93visible_spectroscopy
    THANK YOU

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