SlideShare a Scribd company logo
1 of 49
• MRS.S.AMIRTHAM
• ASSISTANT PROFESSOR
• PG & RESEARCH DEPARTMENT OF BIOTECHNOLOGY
• BON SECOURS COLLEGE FOR WOMEN
• THANJAVUR.
SPECTROSCO
PY
CONTENT
• Beer- Lambert law
• Light absorption and Transmission
• Extinction coefficient
• Application of UV-visible spectroscopic techniques
• Flame photometry
• Atomic absorption spectrophotometey
• Circular Dichroism
• Optical rotatory dispersion
• Principle and application of NMR and ESR techniques
BEER-LAW
• When monochromatic light passes through an absorbing
medium its intensity decreases exponentially as the
concentration of the absorption medium increases.
I = I° e- K2 C
LAMBERT LAW
• When a ray of monochromatic light passes through the
absorbing medium its intensity decreases exponentially as
the length of the absorbing medium increases.
I = I° e- k1 l
LIMITATION OF BEER-LAMBERT LAW
•Only applicable to monochromatic radiations.
•Non-linearity arises at high concentration.
LIGHT ABSORPTION
•Definition
The phenomenon in which a medium
decreases the intensity of the light beam going
through it is called light absorption.
•The law of light absorption
The intensity of a monochromatic light beam
after going through an absorbing medium decreases
as an exponential function of the distance.
•The selective absorption of light
Different light are absorbed
differently. In other words, the absorption
of light through a medium is selective.
TRANSMISSION
Transmission of light is when light
passes through an object depend on whether it
is
• Transparent
• Opaque
• Translucent
TRANSPARENT
• The transparent material
allow light to pass through
with almost no distance.
• It may or may not colour
the light but clearly see
objects through them
Eg: Glass
TRANSLUCENT
• Translucent material allow
only part of the light to pass
through, while also bouncing
it in many new directions.
• It gives you a blurry view
but lets in light.
Eg: A frosted glass door
(like a shower door)
OPAQUE
• Opaque material completely
block light from passing.
Eg: Dark coloured
cardstock paper
IMPORTANCE OF MOLAR EXTINCTION
COEFFICIENT
•Molar extinction coefficient is a measurement of how
strong a chemical species absorb light at a given
wavelength.
•The SI unit are m2 / mol but in practice they are
usually taken as M -1 C -1
•Extinction coefficient allow us for estimation of molar
concentration of solution from its measured
absorbance.
HOW TO FIND MOLAR EXTINCTION
COEFFICIENT
•Beer Lambert law use to find the molar extinction
coefficient.
•The simple way to calculate it using the given
formula.
APPLICATION OF UV-VISIBLE
SPECTROSCOPY
•Detection of impurities.
Uv absorption spectroscopy is one of the best
methods for determining of impurities in organic
methods. By also measuring the absorbance at specific
wavelength, the impurities can be detected.
•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 hetro atoms.
•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 as follows.
•Qualitative analysis.
Uv absorption spectroscopy can characterize
those types of compounds which absorb uv radiation.
•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.
•Detection of fundamental group.
To detect the presence or absence of function
group in the compound
Absence of a band at particular wavelength
regarded as an evidence for the absence of particular
•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 the drugs in a solvent.
•Examinationof 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
polynuclear compounds.
•As HPLC detector.
A Uv /vis spectrophotometer may be used as
a detector for HPLC.
FLAME PHOTOMETRY
•Flame photometry is also called Flame Atomic
Emission Spectrometry is a branch of spectroscopy
in which the species examined in the spectrometer.
BASIC CONCEPTS
• Liquid sample containing
metal salt solution is
introduced into a flame.
• Solvent is first vaporized,
leaving particles of solid salt
which is then vaporized into
gaseous state.
• Gaseous molecule dissociate
to give neutral atoms.
APPLICATION
•Flame photometry has both quantitative and
qualitative applications.
•To estimate sodium, potassium, calcium, lithium etc.
•Flame photometry with mono-chromators emit
radiation of characteristic wavelength which help to
detect the presence of a particular metal in a
sample.
PRINCIPLES
•When a solution is aspirated into a low temperature
flame, in an aerosol ,each droplet of water
evaporates leaving a solid of the residue of
evaporation.
•Various metals emit a characteristic colour of light
when heated.
ATOMIC ABSORPTION
SPECTROSCOPY
•Atomic absorption spectroscopy is a very
common technique for detecting metals and
metallic in samples.
HISTORY
•First atomic absorption spectroscopy was built by
CSIRO scientist,Alan walsh in 1954, shown in the
picture below.
PRINCIPLES
•The technique used basically the principles that free
atoms(gas)generated in an atomizer can absorb
radiation at specific frequency.
•Atomic absorption spectroscopy qualifies the
absorption of ground state atoms in the gaseous
state.
•The atoms absorb ul4or visible light and make
transitions to higher electronic energy levels.
APPLICATION
•Determination of even small amounts of
metals(lead,mercury, calcium, magnesium etc.)
•Environmental studies:drinking water,ocean water,
soil.
•Food industry
•Pharmaceutical industry.
CIRCULAR DICHROISM
•Circular Dichroism spectroscopy is a form of light
absorption spectroscopy that measures the
difference in absorbance of right-left circularly
polarized light.
APPLICATION
•Circular Dichroism phenomena that result when a
symmetrical molecules interact with plane polarized
light.
OPTICAL ROTATORY
DISPERSION
• Optical rotatory dispersion is
the variation in the optical
rotation of a substance with a
change in the wavelength of
light.
FUNDAMENTAL OF ORD
•Plane / linearly polarized light.
•Optical activity.
•Specific rotation.
•Circular Birefringence / optical rotation.
INSTRUMENTATION OF
ORD
• Light source
• Monochromater
• Polarizer
• Analyzer
• Sample tube
• Photo multiplier
APPLICATION
•Quantitative analysis: Specific rotation is a good
measure of concentration.
•Determination of absolute configuration.
Configuration studies. Eg: (+) 3 methyl
cyclohexanone.
PRINCIPLES OF ESR
•The study of the behaviour of the electrons in a
given information about the condition of the sample.
•ESR is used to observe and measure the absorption
of microwave energy by unpaired electrons in a
magnetic field.
APPLICATION OF ESR
•ESR /EPR spectroscopy is used in various branches
of science, such as biology, physics, chemistry, for
the detection and identification of free radicals and
para magnetic centres such as F-centers.
•EPR is a sensitive, specific method for studying both
radicals formed in chemical reaction and the
reaction themselves.
PRINCIPLES OF NMR
•The principles behind NMR is that many
nuclei have spin and all nuclei are
electrically charged. If an external
transfer is Possible between the base
energy to a higher energy level.
APPLICATION OF NMR
•Nuclear magnetic resonance spectroscopy is widely
used to determine the structure of organic molecules
in solution and study molecular physics, crystal as
well as non-crystalline materials.
•NMR is also routinely used in advanced medical
imaging techniques, such as in magnetic resonance
imaging (MRI).
Spectroscopy   amirtham

More Related Content

What's hot

Nephlometry & Turbidometry
Nephlometry & TurbidometryNephlometry & Turbidometry
Nephlometry & Turbidometryrdeepthi1
 
UV VISIBLE REFLECTANCESPECTROSCOPY AND APPLICATION
UV VISIBLE REFLECTANCESPECTROSCOPY AND APPLICATION UV VISIBLE REFLECTANCESPECTROSCOPY AND APPLICATION
UV VISIBLE REFLECTANCESPECTROSCOPY AND APPLICATION OORATHI SASIVARDHAN
 
Photometry and spectrophotometry
Photometry and spectrophotometry Photometry and spectrophotometry
Photometry and spectrophotometry AnumoluRamyasri
 
Infrared spectroscopy
Infrared spectroscopy Infrared spectroscopy
Infrared spectroscopy Prachi Pathak
 
Confocal microscopy Likhith K
Confocal microscopy Likhith KConfocal microscopy Likhith K
Confocal microscopy Likhith KLIKHITHK1
 
Flame photometer
Flame photometerFlame photometer
Flame photometerAown Rizvi
 
Radioisotope technique and methods
Radioisotope technique and methodsRadioisotope technique and methods
Radioisotope technique and methodsUniversity of Mumbai
 
I R spectroscopy & its application
I R spectroscopy & its application I R spectroscopy & its application
I R spectroscopy & its application Amit Agnihotri
 
Deviations from Beers law
Deviations from Beers lawDeviations from Beers law
Deviations from Beers lawVrushali Tambe
 
Instrumentation of uv spectrophotometer pranav kumar singh
Instrumentation of uv spectrophotometer   pranav kumar singhInstrumentation of uv spectrophotometer   pranav kumar singh
Instrumentation of uv spectrophotometer pranav kumar singhAkanksha Mishra
 
Difference in raman spectroscopy and ir spectroscopy
Difference in raman spectroscopy and ir spectroscopyDifference in raman spectroscopy and ir spectroscopy
Difference in raman spectroscopy and ir spectroscopyVrushali Tambe
 

What's hot (19)

Nephlometry & Turbidometry
Nephlometry & TurbidometryNephlometry & Turbidometry
Nephlometry & Turbidometry
 
UV VISIBLE REFLECTANCESPECTROSCOPY AND APPLICATION
UV VISIBLE REFLECTANCESPECTROSCOPY AND APPLICATION UV VISIBLE REFLECTANCESPECTROSCOPY AND APPLICATION
UV VISIBLE REFLECTANCESPECTROSCOPY AND APPLICATION
 
Photometry and spectrophotometry
Photometry and spectrophotometry Photometry and spectrophotometry
Photometry and spectrophotometry
 
Analytical Spectroscopic systems
Analytical Spectroscopic systemsAnalytical Spectroscopic systems
Analytical Spectroscopic systems
 
Photometry
PhotometryPhotometry
Photometry
 
Spectrofluorimetry
SpectrofluorimetrySpectrofluorimetry
Spectrofluorimetry
 
Uv visiblr spectroscopy
Uv visiblr spectroscopyUv visiblr spectroscopy
Uv visiblr spectroscopy
 
Basic principle of spectrophotometry & colorimetry
Basic principle of spectrophotometry & colorimetryBasic principle of spectrophotometry & colorimetry
Basic principle of spectrophotometry & colorimetry
 
NIR ppt
NIR  pptNIR  ppt
NIR ppt
 
Infrared spectroscopy
Infrared spectroscopy Infrared spectroscopy
Infrared spectroscopy
 
Confocal microscopy Likhith K
Confocal microscopy Likhith KConfocal microscopy Likhith K
Confocal microscopy Likhith K
 
Flame photometer
Flame photometerFlame photometer
Flame photometer
 
Infrared spectroscopy
Infrared spectroscopyInfrared spectroscopy
Infrared spectroscopy
 
Radioisotope technique and methods
Radioisotope technique and methodsRadioisotope technique and methods
Radioisotope technique and methods
 
I R spectroscopy & its application
I R spectroscopy & its application I R spectroscopy & its application
I R spectroscopy & its application
 
Flame emission spectroscopy
Flame emission spectroscopyFlame emission spectroscopy
Flame emission spectroscopy
 
Deviations from Beers law
Deviations from Beers lawDeviations from Beers law
Deviations from Beers law
 
Instrumentation of uv spectrophotometer pranav kumar singh
Instrumentation of uv spectrophotometer   pranav kumar singhInstrumentation of uv spectrophotometer   pranav kumar singh
Instrumentation of uv spectrophotometer pranav kumar singh
 
Difference in raman spectroscopy and ir spectroscopy
Difference in raman spectroscopy and ir spectroscopyDifference in raman spectroscopy and ir spectroscopy
Difference in raman spectroscopy and ir spectroscopy
 

Similar to Spectroscopy amirtham

Spectroscopy techniques, it's principle, types and applications
Spectroscopy techniques, it's principle, types and applications Spectroscopy techniques, it's principle, types and applications
Spectroscopy techniques, it's principle, types and applications NizadSultana
 
Spectrophotometry.pptx
Spectrophotometry.pptxSpectrophotometry.pptx
Spectrophotometry.pptxRafeeqCM1
 
Spectrophotometry Shrraddha suman.pptx
Spectrophotometry Shrraddha suman.pptxSpectrophotometry Shrraddha suman.pptx
Spectrophotometry Shrraddha suman.pptxShrraddha
 
Photometry, colorimetry, spectophotometry
Photometry, colorimetry, spectophotometryPhotometry, colorimetry, spectophotometry
Photometry, colorimetry, spectophotometrygohil sanjay bhagvanji
 
ATOMIC ABSORPTION SPECTROSCOPY.pptx
ATOMIC ABSORPTION SPECTROSCOPY.pptxATOMIC ABSORPTION SPECTROSCOPY.pptx
ATOMIC ABSORPTION SPECTROSCOPY.pptxShivamSharma613110
 
SPECTROCOPY PRESENTATION.pptx
SPECTROCOPY PRESENTATION.pptxSPECTROCOPY PRESENTATION.pptx
SPECTROCOPY PRESENTATION.pptxLazarusJoseph5
 
Atomic absorption spectroscopy
Atomic absorption spectroscopyAtomic absorption spectroscopy
Atomic absorption spectroscopySeenam Iftikhar
 
Principle and instrumentation
Principle and instrumentation Principle and instrumentation
Principle and instrumentation ShivaniPatil68
 
NEPHLOMETRY and TURBIDIMETRY PPT 1
NEPHLOMETRY and TURBIDIMETRY PPT 1NEPHLOMETRY and TURBIDIMETRY PPT 1
NEPHLOMETRY and TURBIDIMETRY PPT 1shaisejacob
 
Atomic emission specrtroscopy for subject Parmaceutical Analysis
Atomic emission specrtroscopy for subject Parmaceutical AnalysisAtomic emission specrtroscopy for subject Parmaceutical Analysis
Atomic emission specrtroscopy for subject Parmaceutical AnalysisVrajPatel208
 
UV Spectrophotometer.pptx
UV Spectrophotometer.pptxUV Spectrophotometer.pptx
UV Spectrophotometer.pptx510Sudha
 
Flourimetry
FlourimetryFlourimetry
FlourimetryAFSATH
 
Carbon dots characterization and applications
Carbon dots characterization and applicationsCarbon dots characterization and applications
Carbon dots characterization and applicationspchandrasekaran
 

Similar to Spectroscopy amirtham (20)

Flame photometer
Flame photometer Flame photometer
Flame photometer
 
spectroscopy-210610191026.pptx
spectroscopy-210610191026.pptxspectroscopy-210610191026.pptx
spectroscopy-210610191026.pptx
 
Spectroscopy techniques, it's principle, types and applications
Spectroscopy techniques, it's principle, types and applications Spectroscopy techniques, it's principle, types and applications
Spectroscopy techniques, it's principle, types and applications
 
Spectrophotometry.pptx
Spectrophotometry.pptxSpectrophotometry.pptx
Spectrophotometry.pptx
 
Spectrophotometry Shrraddha suman.pptx
Spectrophotometry Shrraddha suman.pptxSpectrophotometry Shrraddha suman.pptx
Spectrophotometry Shrraddha suman.pptx
 
Photometry, colorimetry, spectophotometry
Photometry, colorimetry, spectophotometryPhotometry, colorimetry, spectophotometry
Photometry, colorimetry, spectophotometry
 
ATOMIC ABSORPTION SPECTROSCOPY.pptx
ATOMIC ABSORPTION SPECTROSCOPY.pptxATOMIC ABSORPTION SPECTROSCOPY.pptx
ATOMIC ABSORPTION SPECTROSCOPY.pptx
 
SPECTROCOPY PRESENTATION.pptx
SPECTROCOPY PRESENTATION.pptxSPECTROCOPY PRESENTATION.pptx
SPECTROCOPY PRESENTATION.pptx
 
Uv visible spectroscopy
Uv visible spectroscopyUv visible spectroscopy
Uv visible spectroscopy
 
UV-VISIBLE SPECTROSCOPY.pptx
UV-VISIBLE SPECTROSCOPY.pptxUV-VISIBLE SPECTROSCOPY.pptx
UV-VISIBLE SPECTROSCOPY.pptx
 
Atomic absorption spectroscopy
Atomic absorption spectroscopyAtomic absorption spectroscopy
Atomic absorption spectroscopy
 
Principle and instrumentation
Principle and instrumentation Principle and instrumentation
Principle and instrumentation
 
NEPHLOMETRY and TURBIDIMETRY PPT 1
NEPHLOMETRY and TURBIDIMETRY PPT 1NEPHLOMETRY and TURBIDIMETRY PPT 1
NEPHLOMETRY and TURBIDIMETRY PPT 1
 
Atomic emission specrtroscopy for subject Parmaceutical Analysis
Atomic emission specrtroscopy for subject Parmaceutical AnalysisAtomic emission specrtroscopy for subject Parmaceutical Analysis
Atomic emission specrtroscopy for subject Parmaceutical Analysis
 
UV Spectroscopy
UV Spectroscopy UV Spectroscopy
UV Spectroscopy
 
5 nephalo turbido jntu pharmacy
5 nephalo turbido jntu pharmacy5 nephalo turbido jntu pharmacy
5 nephalo turbido jntu pharmacy
 
UV Spectrophotometer.pptx
UV Spectrophotometer.pptxUV Spectrophotometer.pptx
UV Spectrophotometer.pptx
 
Flourimetry
FlourimetryFlourimetry
Flourimetry
 
Carbon dots characterization and applications
Carbon dots characterization and applicationsCarbon dots characterization and applications
Carbon dots characterization and applications
 
Colorimetry
ColorimetryColorimetry
Colorimetry
 

More from christanantony

More from christanantony (20)

Pcr molecular diagnosis
Pcr molecular diagnosisPcr molecular diagnosis
Pcr molecular diagnosis
 
Pcr and its application
Pcr and its applicationPcr and its application
Pcr and its application
 
Gene silencing amirtham
Gene silencing   amirthamGene silencing   amirtham
Gene silencing amirtham
 
Cloning pcr amirtham
Cloning pcr   amirthamCloning pcr   amirtham
Cloning pcr amirtham
 
C dna and genomic libraries amirtham
C dna and genomic libraries   amirthamC dna and genomic libraries   amirtham
C dna and genomic libraries amirtham
 
Electrophoresis amirtham
Electrophoresis   amirthamElectrophoresis   amirtham
Electrophoresis amirtham
 
Crystallography amirtham
Crystallography   amirthamCrystallography   amirtham
Crystallography amirtham
 
Chromatography amirtham
Chromatography   amirthamChromatography   amirtham
Chromatography amirtham
 
Centrifugation amirtham
Centrifugation   amirthamCentrifugation   amirtham
Centrifugation amirtham
 
Calorimeter
CalorimeterCalorimeter
Calorimeter
 
C dna and genomic libraries copy
C dna and genomic libraries   copyC dna and genomic libraries   copy
C dna and genomic libraries copy
 
Daisy gene 123
Daisy gene  123Daisy gene  123
Daisy gene 123
 
Presentation
Presentation Presentation
Presentation
 
Cloning pcr
Cloning pcrCloning pcr
Cloning pcr
 
Labelling of dna
Labelling of dnaLabelling of dna
Labelling of dna
 
Spectroscopy
SpectroscopySpectroscopy
Spectroscopy
 
Electrophoresis wps office
Electrophoresis wps officeElectrophoresis wps office
Electrophoresis wps office
 
Crystallography
CrystallographyCrystallography
Crystallography
 
Centrifugation
Centrifugation Centrifugation
Centrifugation
 
Centrifugation by akasthiya
Centrifugation  by akasthiyaCentrifugation  by akasthiya
Centrifugation by akasthiya
 

Recently uploaded

Triploidy ...............................pptx
Triploidy ...............................pptxTriploidy ...............................pptx
Triploidy ...............................pptxCherry
 
Plasmapheresis - Dr. E. Muralinath - Kalyan . C.pptx
Plasmapheresis - Dr. E. Muralinath - Kalyan . C.pptxPlasmapheresis - Dr. E. Muralinath - Kalyan . C.pptx
Plasmapheresis - Dr. E. Muralinath - Kalyan . C.pptxmuralinath2
 
family therapy psychotherapy types .pdf
family therapy psychotherapy types  .pdffamily therapy psychotherapy types  .pdf
family therapy psychotherapy types .pdfhaseebahmeddrama
 
Gliese 12 b: A Temperate Earth-sized Planet at 12 pc Ideal for Atmospheric Tr...
Gliese 12 b: A Temperate Earth-sized Planet at 12 pc Ideal for Atmospheric Tr...Gliese 12 b: A Temperate Earth-sized Planet at 12 pc Ideal for Atmospheric Tr...
Gliese 12 b: A Temperate Earth-sized Planet at 12 pc Ideal for Atmospheric Tr...Sérgio Sacani
 
GBSN - Microbiology (Unit 7) Microbiology in Everyday Life
GBSN - Microbiology (Unit 7) Microbiology in Everyday LifeGBSN - Microbiology (Unit 7) Microbiology in Everyday Life
GBSN - Microbiology (Unit 7) Microbiology in Everyday LifeAreesha Ahmad
 
The Scientific names of some important families of Industrial plants .pdf
The Scientific names of some important families of Industrial plants .pdfThe Scientific names of some important families of Industrial plants .pdf
The Scientific names of some important families of Industrial plants .pdfMohamed Said
 
The importance of continents, oceans and plate tectonics for the evolution of...
The importance of continents, oceans and plate tectonics for the evolution of...The importance of continents, oceans and plate tectonics for the evolution of...
The importance of continents, oceans and plate tectonics for the evolution of...Sérgio Sacani
 
GBSN - Microbiology Lab 1 (Microbiology Lab Safety Procedures)
GBSN -  Microbiology Lab  1 (Microbiology Lab Safety Procedures)GBSN -  Microbiology Lab  1 (Microbiology Lab Safety Procedures)
GBSN - Microbiology Lab 1 (Microbiology Lab Safety Procedures)Areesha Ahmad
 
Cell Immobilization Methods and Applications.pptx
Cell Immobilization Methods and Applications.pptxCell Immobilization Methods and Applications.pptx
Cell Immobilization Methods and Applications.pptxCherry
 
Lec 1.b Totipotency and birth of tissue culture.ppt
Lec 1.b Totipotency and birth of tissue culture.pptLec 1.b Totipotency and birth of tissue culture.ppt
Lec 1.b Totipotency and birth of tissue culture.pptasifali1111
 
GBSN - Biochemistry (Unit 4) Chemistry of Carbohydrates
GBSN - Biochemistry (Unit 4) Chemistry of CarbohydratesGBSN - Biochemistry (Unit 4) Chemistry of Carbohydrates
GBSN - Biochemistry (Unit 4) Chemistry of CarbohydratesAreesha Ahmad
 
WASP-69b’s Escaping Envelope Is Confined to a Tail Extending at Least 7 Rp
WASP-69b’s Escaping Envelope Is Confined to a Tail Extending at Least 7 RpWASP-69b’s Escaping Envelope Is Confined to a Tail Extending at Least 7 Rp
WASP-69b’s Escaping Envelope Is Confined to a Tail Extending at Least 7 RpSérgio Sacani
 
Emergent ribozyme behaviors in oxychlorine brines indicate a unique niche for...
Emergent ribozyme behaviors in oxychlorine brines indicate a unique niche for...Emergent ribozyme behaviors in oxychlorine brines indicate a unique niche for...
Emergent ribozyme behaviors in oxychlorine brines indicate a unique niche for...Sérgio Sacani
 
Mitosis...............................pptx
Mitosis...............................pptxMitosis...............................pptx
Mitosis...............................pptxCherry
 
Continuum emission from within the plunging region of black hole discs
Continuum emission from within the plunging region of black hole discsContinuum emission from within the plunging region of black hole discs
Continuum emission from within the plunging region of black hole discsSérgio Sacani
 
Ostiguy & Panizza & Moffitt (eds.) - Populism in Global Perspective. A Perfor...
Ostiguy & Panizza & Moffitt (eds.) - Populism in Global Perspective. A Perfor...Ostiguy & Panizza & Moffitt (eds.) - Populism in Global Perspective. A Perfor...
Ostiguy & Panizza & Moffitt (eds.) - Populism in Global Perspective. A Perfor...frank0071
 
Tuberculosis (TB)-Notes.pdf microbiology notes
Tuberculosis (TB)-Notes.pdf microbiology notesTuberculosis (TB)-Notes.pdf microbiology notes
Tuberculosis (TB)-Notes.pdf microbiology notesjyothisaisri
 
Aerodynamics. flippatterncn5tm5ttnj6nmnynyppt
Aerodynamics. flippatterncn5tm5ttnj6nmnynypptAerodynamics. flippatterncn5tm5ttnj6nmnynyppt
Aerodynamics. flippatterncn5tm5ttnj6nmnynypptsreddyrahul
 
MODERN PHYSICS_REPORTING_QUANTA_.....pdf
MODERN PHYSICS_REPORTING_QUANTA_.....pdfMODERN PHYSICS_REPORTING_QUANTA_.....pdf
MODERN PHYSICS_REPORTING_QUANTA_.....pdfRevenJadePalma
 
SCHISTOSOMA HEAMATOBIUM life cycle .pdf
SCHISTOSOMA HEAMATOBIUM life cycle  .pdfSCHISTOSOMA HEAMATOBIUM life cycle  .pdf
SCHISTOSOMA HEAMATOBIUM life cycle .pdfDebdattaGhosh6
 

Recently uploaded (20)

Triploidy ...............................pptx
Triploidy ...............................pptxTriploidy ...............................pptx
Triploidy ...............................pptx
 
Plasmapheresis - Dr. E. Muralinath - Kalyan . C.pptx
Plasmapheresis - Dr. E. Muralinath - Kalyan . C.pptxPlasmapheresis - Dr. E. Muralinath - Kalyan . C.pptx
Plasmapheresis - Dr. E. Muralinath - Kalyan . C.pptx
 
family therapy psychotherapy types .pdf
family therapy psychotherapy types  .pdffamily therapy psychotherapy types  .pdf
family therapy psychotherapy types .pdf
 
Gliese 12 b: A Temperate Earth-sized Planet at 12 pc Ideal for Atmospheric Tr...
Gliese 12 b: A Temperate Earth-sized Planet at 12 pc Ideal for Atmospheric Tr...Gliese 12 b: A Temperate Earth-sized Planet at 12 pc Ideal for Atmospheric Tr...
Gliese 12 b: A Temperate Earth-sized Planet at 12 pc Ideal for Atmospheric Tr...
 
GBSN - Microbiology (Unit 7) Microbiology in Everyday Life
GBSN - Microbiology (Unit 7) Microbiology in Everyday LifeGBSN - Microbiology (Unit 7) Microbiology in Everyday Life
GBSN - Microbiology (Unit 7) Microbiology in Everyday Life
 
The Scientific names of some important families of Industrial plants .pdf
The Scientific names of some important families of Industrial plants .pdfThe Scientific names of some important families of Industrial plants .pdf
The Scientific names of some important families of Industrial plants .pdf
 
The importance of continents, oceans and plate tectonics for the evolution of...
The importance of continents, oceans and plate tectonics for the evolution of...The importance of continents, oceans and plate tectonics for the evolution of...
The importance of continents, oceans and plate tectonics for the evolution of...
 
GBSN - Microbiology Lab 1 (Microbiology Lab Safety Procedures)
GBSN -  Microbiology Lab  1 (Microbiology Lab Safety Procedures)GBSN -  Microbiology Lab  1 (Microbiology Lab Safety Procedures)
GBSN - Microbiology Lab 1 (Microbiology Lab Safety Procedures)
 
Cell Immobilization Methods and Applications.pptx
Cell Immobilization Methods and Applications.pptxCell Immobilization Methods and Applications.pptx
Cell Immobilization Methods and Applications.pptx
 
Lec 1.b Totipotency and birth of tissue culture.ppt
Lec 1.b Totipotency and birth of tissue culture.pptLec 1.b Totipotency and birth of tissue culture.ppt
Lec 1.b Totipotency and birth of tissue culture.ppt
 
GBSN - Biochemistry (Unit 4) Chemistry of Carbohydrates
GBSN - Biochemistry (Unit 4) Chemistry of CarbohydratesGBSN - Biochemistry (Unit 4) Chemistry of Carbohydrates
GBSN - Biochemistry (Unit 4) Chemistry of Carbohydrates
 
WASP-69b’s Escaping Envelope Is Confined to a Tail Extending at Least 7 Rp
WASP-69b’s Escaping Envelope Is Confined to a Tail Extending at Least 7 RpWASP-69b’s Escaping Envelope Is Confined to a Tail Extending at Least 7 Rp
WASP-69b’s Escaping Envelope Is Confined to a Tail Extending at Least 7 Rp
 
Emergent ribozyme behaviors in oxychlorine brines indicate a unique niche for...
Emergent ribozyme behaviors in oxychlorine brines indicate a unique niche for...Emergent ribozyme behaviors in oxychlorine brines indicate a unique niche for...
Emergent ribozyme behaviors in oxychlorine brines indicate a unique niche for...
 
Mitosis...............................pptx
Mitosis...............................pptxMitosis...............................pptx
Mitosis...............................pptx
 
Continuum emission from within the plunging region of black hole discs
Continuum emission from within the plunging region of black hole discsContinuum emission from within the plunging region of black hole discs
Continuum emission from within the plunging region of black hole discs
 
Ostiguy & Panizza & Moffitt (eds.) - Populism in Global Perspective. A Perfor...
Ostiguy & Panizza & Moffitt (eds.) - Populism in Global Perspective. A Perfor...Ostiguy & Panizza & Moffitt (eds.) - Populism in Global Perspective. A Perfor...
Ostiguy & Panizza & Moffitt (eds.) - Populism in Global Perspective. A Perfor...
 
Tuberculosis (TB)-Notes.pdf microbiology notes
Tuberculosis (TB)-Notes.pdf microbiology notesTuberculosis (TB)-Notes.pdf microbiology notes
Tuberculosis (TB)-Notes.pdf microbiology notes
 
Aerodynamics. flippatterncn5tm5ttnj6nmnynyppt
Aerodynamics. flippatterncn5tm5ttnj6nmnynypptAerodynamics. flippatterncn5tm5ttnj6nmnynyppt
Aerodynamics. flippatterncn5tm5ttnj6nmnynyppt
 
MODERN PHYSICS_REPORTING_QUANTA_.....pdf
MODERN PHYSICS_REPORTING_QUANTA_.....pdfMODERN PHYSICS_REPORTING_QUANTA_.....pdf
MODERN PHYSICS_REPORTING_QUANTA_.....pdf
 
SCHISTOSOMA HEAMATOBIUM life cycle .pdf
SCHISTOSOMA HEAMATOBIUM life cycle  .pdfSCHISTOSOMA HEAMATOBIUM life cycle  .pdf
SCHISTOSOMA HEAMATOBIUM life cycle .pdf
 

Spectroscopy amirtham

  • 1. • MRS.S.AMIRTHAM • ASSISTANT PROFESSOR • PG & RESEARCH DEPARTMENT OF BIOTECHNOLOGY • BON SECOURS COLLEGE FOR WOMEN • THANJAVUR.
  • 2.
  • 4. CONTENT • Beer- Lambert law • Light absorption and Transmission • Extinction coefficient • Application of UV-visible spectroscopic techniques • Flame photometry • Atomic absorption spectrophotometey • Circular Dichroism
  • 5. • Optical rotatory dispersion • Principle and application of NMR and ESR techniques
  • 6. BEER-LAW • When monochromatic light passes through an absorbing medium its intensity decreases exponentially as the concentration of the absorption medium increases. I = I° e- K2 C
  • 7. LAMBERT LAW • When a ray of monochromatic light passes through the absorbing medium its intensity decreases exponentially as the length of the absorbing medium increases. I = I° e- k1 l
  • 8. LIMITATION OF BEER-LAMBERT LAW •Only applicable to monochromatic radiations. •Non-linearity arises at high concentration.
  • 9. LIGHT ABSORPTION •Definition The phenomenon in which a medium decreases the intensity of the light beam going through it is called light absorption. •The law of light absorption The intensity of a monochromatic light beam after going through an absorbing medium decreases as an exponential function of the distance.
  • 10. •The selective absorption of light Different light are absorbed differently. In other words, the absorption of light through a medium is selective.
  • 11.
  • 12. TRANSMISSION Transmission of light is when light passes through an object depend on whether it is • Transparent • Opaque • Translucent
  • 13. TRANSPARENT • The transparent material allow light to pass through with almost no distance. • It may or may not colour the light but clearly see objects through them Eg: Glass
  • 14. TRANSLUCENT • Translucent material allow only part of the light to pass through, while also bouncing it in many new directions. • It gives you a blurry view but lets in light. Eg: A frosted glass door (like a shower door)
  • 15. OPAQUE • Opaque material completely block light from passing. Eg: Dark coloured cardstock paper
  • 16. IMPORTANCE OF MOLAR EXTINCTION COEFFICIENT •Molar extinction coefficient is a measurement of how strong a chemical species absorb light at a given wavelength. •The SI unit are m2 / mol but in practice they are usually taken as M -1 C -1 •Extinction coefficient allow us for estimation of molar concentration of solution from its measured absorbance.
  • 17. HOW TO FIND MOLAR EXTINCTION COEFFICIENT •Beer Lambert law use to find the molar extinction coefficient. •The simple way to calculate it using the given formula.
  • 18. APPLICATION OF UV-VISIBLE SPECTROSCOPY •Detection of impurities. Uv absorption spectroscopy is one of the best methods for determining of impurities in organic methods. By also measuring the absorbance at specific wavelength, the impurities can be detected.
  • 19.
  • 20. •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 hetro atoms. •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 as follows.
  • 21.
  • 22. •Qualitative analysis. Uv absorption spectroscopy can characterize those types of compounds which absorb uv radiation.
  • 23. •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. •Detection of fundamental group. To detect the presence or absence of function group in the compound Absence of a band at particular wavelength regarded as an evidence for the absence of particular
  • 24. •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 the drugs in a solvent. •Examinationof 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 polynuclear compounds.
  • 25. •As HPLC detector. A Uv /vis spectrophotometer may be used as a detector for HPLC.
  • 26. FLAME PHOTOMETRY •Flame photometry is also called Flame Atomic Emission Spectrometry is a branch of spectroscopy in which the species examined in the spectrometer.
  • 27. BASIC CONCEPTS • Liquid sample containing metal salt solution is introduced into a flame. • Solvent is first vaporized, leaving particles of solid salt which is then vaporized into gaseous state. • Gaseous molecule dissociate to give neutral atoms.
  • 28. APPLICATION •Flame photometry has both quantitative and qualitative applications. •To estimate sodium, potassium, calcium, lithium etc. •Flame photometry with mono-chromators emit radiation of characteristic wavelength which help to detect the presence of a particular metal in a sample.
  • 29. PRINCIPLES •When a solution is aspirated into a low temperature flame, in an aerosol ,each droplet of water evaporates leaving a solid of the residue of evaporation. •Various metals emit a characteristic colour of light when heated.
  • 30.
  • 31. ATOMIC ABSORPTION SPECTROSCOPY •Atomic absorption spectroscopy is a very common technique for detecting metals and metallic in samples.
  • 32.
  • 33. HISTORY •First atomic absorption spectroscopy was built by CSIRO scientist,Alan walsh in 1954, shown in the picture below.
  • 34. PRINCIPLES •The technique used basically the principles that free atoms(gas)generated in an atomizer can absorb radiation at specific frequency. •Atomic absorption spectroscopy qualifies the absorption of ground state atoms in the gaseous state. •The atoms absorb ul4or visible light and make transitions to higher electronic energy levels.
  • 35.
  • 36. APPLICATION •Determination of even small amounts of metals(lead,mercury, calcium, magnesium etc.) •Environmental studies:drinking water,ocean water, soil. •Food industry •Pharmaceutical industry.
  • 37. CIRCULAR DICHROISM •Circular Dichroism spectroscopy is a form of light absorption spectroscopy that measures the difference in absorbance of right-left circularly polarized light.
  • 38.
  • 39.
  • 40. APPLICATION •Circular Dichroism phenomena that result when a symmetrical molecules interact with plane polarized light.
  • 41. OPTICAL ROTATORY DISPERSION • Optical rotatory dispersion is the variation in the optical rotation of a substance with a change in the wavelength of light.
  • 42. FUNDAMENTAL OF ORD •Plane / linearly polarized light. •Optical activity. •Specific rotation. •Circular Birefringence / optical rotation.
  • 43. INSTRUMENTATION OF ORD • Light source • Monochromater • Polarizer • Analyzer • Sample tube • Photo multiplier
  • 44. APPLICATION •Quantitative analysis: Specific rotation is a good measure of concentration. •Determination of absolute configuration. Configuration studies. Eg: (+) 3 methyl cyclohexanone.
  • 45. PRINCIPLES OF ESR •The study of the behaviour of the electrons in a given information about the condition of the sample. •ESR is used to observe and measure the absorption of microwave energy by unpaired electrons in a magnetic field.
  • 46. APPLICATION OF ESR •ESR /EPR spectroscopy is used in various branches of science, such as biology, physics, chemistry, for the detection and identification of free radicals and para magnetic centres such as F-centers. •EPR is a sensitive, specific method for studying both radicals formed in chemical reaction and the reaction themselves.
  • 47. PRINCIPLES OF NMR •The principles behind NMR is that many nuclei have spin and all nuclei are electrically charged. If an external transfer is Possible between the base energy to a higher energy level.
  • 48. APPLICATION OF NMR •Nuclear magnetic resonance spectroscopy is widely used to determine the structure of organic molecules in solution and study molecular physics, crystal as well as non-crystalline materials. •NMR is also routinely used in advanced medical imaging techniques, such as in magnetic resonance imaging (MRI).