Spectral studies

• Measurement of radiant energy absorbed,

emitted, transmitted by the molecules
Atomic spectroscopy
• Change in energy take place at atomic level
• Atomic absorption spectra & Flame emission
spectra
Molecular spectroscopy
• Changes in energy takes place in molecular level
• UV spectroscopy, Colorimetry, IR, Fluorimetry
 Classification

Absorbtion spectra
• Absorbtion of radiation :- U-V, IR,
Colorimetry, NMR & AAS.
Emission Spectra
• Emission of radiation :- Flame, Flourimetry,
Electromagnetic Radiation
• Under the influence of magnetic field :- NMR
 Visible Spectra
Photometer
• Measure the sum of two radiant power of two
electro magnetic beams
Spectro Photometer
• Measure the radiant of two elctro magnetic
waves over a larger wave length
Colorimeter
• Measure the absorbtion in the visible region.
 Applications
• Measure the • Phenolic compounds –
absorption in the Ferric chloride
visible region • Digitoxin – Alkaline
• 400 – 800nm picric acid – 495nm
• Percentage purity • Ergometrine – P-
• Comparing absorption dimethyl amino
spectra benzaldehyde –
• Determination of 545nm
molecular weight • Anthroquinone – Alkali
• Qualitative and – 505nm
Quantitative analysis
 U-V Spectra
• Electron transition from ground level to
excited level.
• Based upon Lamberts & Beers Law
• 200 – 400nm
• Detection of impurities
• Detection of reserpine (268nm), Morphine
(286nm), Vaniline (301nm).
Structural elucidation :-
• Presence of unsaturation,
• presence of hetero atoms, S, O,N.
 IR Spectra
• Study of reflected, absorbed, transmitted radiant
energy
• 2500nm to 16000nm
• Quanitative analysis of antibiotics, alkaloids &
steroidal saponins.
• Identification of impurities
Structural elucidation :-
• Identification of functional group :- Amino group,
OH group..
• Determination of hydrogen bonds
• Region below 1200cm-1 is called finger print
 FTIR
• FTIR is most useful for identifying
phytoconstituents from medicinal plants.
• The term Fourier Transform Infrared
Spectroscopy (FTIR)
• FTIR is perhaps the most powerful tool for
identifying types of chemical bonds and
functional groups.
• Molecular bonds vibrate at various
frequencies depending on the elements and
the type of bonds. For any given bond, there
are several specific frequencies at which it
can vibrate.
Sample Preparation
• The sample was prepared by KBr pellet
method.
• The isolated compounds were milled with
potassium bromide (KBr) to form a very
fine powder.
• This powder was then compressed into a
thin pellet which can be analyzed.
• KBr is also transparent in the IR.
 NMR Spectra
• Measure the absorption of radiation
frequency in the magnetic field
• Structural elucidation of organic
compounds
• Types of proton – Aliphatic, Aromatic &
number of proton
• Number of Hydrogen atoms or Carbon
atoms
• 1H NMR & 13C NMR
 NMR Spectra
• The types of atoms present in the sample
• The relative amounts of atoms present in a
sample
• The specific environments of atoms within
a molecule
• The purity and composition of a sample
• Structural information about a molecule,
including constitutional and conformational
isomerisation
 MASS Spectra

• Molecules are bombarded with electron

• Electrons are ionized & fragmentation of
molecules takes place
• Determination of mass to charge ratio (m/e)
• Neutral molecule - +ve charged ion
• Structural elucidation
• Molecular weight & Molecular formula
 Fluorescence Analysis
• Fluorimetry
Substance absorb light &
reemit radiation • Qualitative & Quantitative
analysis
• Cinchona bark – Yellow line
with blue X RAY Diffraction
• Rhubarb :- R. emodi – brown, • Crystalline nature of compounds
R. rhoponticum – violet • Physical method of structural
• Sesame oil – blue determination
• Berberine – Yellow
• Ergot – Red
• Olive oil – Deep golden yellow
• Arachys oil – blue
• Genetain root – light blue
• Aconite – light blue
• Emetine - Orange