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Seminar on Applications of U.V.
                Spectroscopy




                   Submitted to:               Submitted by:
               Mr. Rambabu Tripathi            Manish Kumar
                                            M.Pharm (Pharmacology)


Department of Pharmacy , Institute of Bio-Medical Education & Research,
               Mangalayatan university Beswan, Aligarh
INTRODUCTION:

   The word spectroscopy implies that we will use the
    electromagnetic spectrum to gain information about
    organic molecules. The modifier ultraviolet means that
    the information will come from a specific region of the
    electromagnetic spectrum called the ultraviolet region
    (190 to 400 nm U.V. Region and 400 to 800 nm
    Visible Region) .
APPLICATIONS OF U.V. SPECTROSCOPY:

1.    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 and it
      can be compared with that of standard raw material. By also
      measuring the absorbance at specific wavelength, the
      impurities can be detected.
U.V. SPECTRA OF PARACETAMOL (PCM)
2. Structure elucidation of organic compounds.


 UV   spectroscopy is useful in the structure elucidation of
 organic molecules, the presence or absence of
 unsaturation, the presence of hetero atoms.


 From   the location of peaks and combination of peaks, it can
 be concluded that whether the compound is saturated or
 unsaturated, hetero atoms are present or not etc.
3. QUANTITATIVE ANALYSIS

   UV absorption spectroscopy can be used for the quantitative
    determination of compounds that absorb UV radiation. This
    determination is based on Beer’s law which is as follows.


              A = log I0 / It = log 1/ T = – log T = abc = εbc
Where :
ε -is extinction co-efficient,
c- is concentration, and
b- is the length of the cell that is used in UV spectrophotometer.
BEER’S LAW
4. QUALITATIVE ANALYSIS

 UV   absorption spectroscopy can characterize those
 types of compounds which absorbs UV radiation.
 Identification is done by comparing the absorption
 spectrum with the spectra of known compounds.
U.V. SPECTRA'S OF IBUPROFEN
5. CHEMICAL KINETICS

   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.
6. DETECTION OF FUNCTIONAL GROUPS

   This technique is used to detect the presence or
    absence of functional group in the compound



   Absence of a band at particular wavelength
    regarded as an evidence for absence of particular
    group
BENZENE   TOLUNE
7. QUANTITATIVE ANALYSIS OF PHARMACEUTICAL
SUBSTANCES



  Many   drugs are either in the form of raw material or
   in the form of formulation. They can be assayed by
   making a suitable solution of the drug in a solvent and
   measuring the absorbance at specific wavelength.


  Diazepam   tablet can be analyzed by 0.5% H2SO4 in
   methanol at the wavelength 284 nm.
8. EXAMINATION OF POLYNUCLEAR HYDROCARBONS


   Benzene and Polynuclear hydrocarbons have characteristic spectra
    in ultraviolet and visible region. Thus identification of
    Polynuclear hydrocarbons can be made by comparison with the
    spectra of known Polynuclear compounds.


   Polynuclear hydrocarbons are the Hydrocarbon molecule with two
    or more closed rings; examples are naphthalene, C10H8, with two
    benzene rings side by side, or diphenyl, (C6H5)2, with two bond-
    connected benzene rings. Also known as polycyclic hydrocarbon.
NAPHTHALENE   DIPHENYL
9. MOLECULAR WEIGHT DETERMINATION

   Molecular    weights    of    compounds      can   be    measured
    spectrophotometrically by preparing the suitable derivatives of
    these compounds.

   For example, if we want to determine the molecular weight of
    amine then it is converted in to amine picrate. Then known
    concentration of amine picrate is dissolved in a litre
    of solution and its optical density is measured at λmax 380 nm.
 After   this the concentration of the solution in gm moles
 per litre can be calculated by using the following formula.




 "c"     can be calculated using above equation, the
 weight       "w"    of    amine     picrate   is    known.
 From "c" and "w", molecular weight of amine picrate can
 be calculated. And the molecular weight of picrate can be
 calculated using the molecular weight of amine picrate.
10. AS HPLC DETECTOR

A   UV/Vis spectrophotometer may be used as a
 detector for HPLC.
REFERENCES:

1.   Sharma. Y.R. Elementary Organic Spectroscopy.
     First edition .S.Chand Publisher; 2010.

2.   Chatwal G.R. Instrumental methods of chemical
     analysis. First edition. Himalaya Publisher; 2010.

3.   www.chem.agilent.com/Library/applications/uv31.pd
     f.

4.   www.uvitron.com/Applications.html.
Application of u.v. spectroscopy