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Optical Rotatory Dispersion &
Circular Dichorism
 Presented by:-
 Sukhjeet kaur
 M.Sc. (H)
biotech(2)
 130181106
CONTENT
 ORD
 PRINCIPLE
 FUNDAMENTAL OF ORD
 CD
 COTTON EFFECT
 ORD CURVES
 APPLICATION
 DIFFERENCE B/W ORD & CD
 CONCLUSION
ORD
 ORD is technique related to the optical
activity
 Rate of change of specific rotation
with change in wave length
 Optical rotation caused by compound
changed with wavelength of light was
first denoted by Biot in 1817
FUNDAMENTAL OF ORD
 Plane polarized light
 Optical activity
 Specific rotation
 Circular birefringence
 Optical rotation
 Elliphicity
Plane polarized light
 Light from ordinary lamp consists of
waves vibrating in many different
planes
 It is passed through Polaroid lens it is
found to be vibrate in one plane is said
to be plane polarized light
Optical rotatory dispersion &                    circular dichorism
OPTICAL ACTIVITY
 The compounds which are having the
ability to rotate the plane of polarized
light are called optical active
compound
 This property of compounds is called
optical activity
 Measured by Polarimeter
 Dextrorotatory
 Levorotatory
Optical rotatory dispersion &                    circular dichorism
SPECIFIC ROTATION
 The specific rotation ([α]) is
an intensive property of a chemical
compound, defined as the change in
orientation of the plane of linearly
polarized light as this light passes
through a sample with a path length of
1 decimeter and a sample
concentration of 1 gram per 1 millilitre
 It is denoted by [α]
CIRCULAR
BIREFRINGENCE
 Birefringence is the optical property
of a material having a refractive
index that depends on
the polarization and propagation
direction of light
 α= (nL-nR)π/ α
CIRCULAR DICHORISM
Some material posses special properties of
absorption of the light circularly light to
different extent than the right circulary
polarized light
Chiral or asymmetric compounds produce
CD spectrum
COTTON EFFECT
This phenomenon
was discovered in
1895 by the French
physicist Aimé
Cotton
Combination of
circular dichorism
& circular
birefringence
ORD CURVES
 Plain curves
 Anomalous curves
 single cotton effect curves
 multiple cotton effect curve
PLAIN CURVES
 Normal smooth curves
 Do not cross the zero rotation line
 e.g.:- alcohols hydrocarbons
ANOMALOUS CURVES
 Shows no. of extreme peaks &
troughs
SCEC & MCEC
 Two or more peaks
& trough
 e.g. ketosteroids
 camphor
APPLICATION
 CD/ORD of protein
 Qualitative analysis of molecules
 Enzyme co-factor evaluation
 CD/ORD of proteins
 CD as finger printing
 CD/ORD of nucleic acids
 CD of B-DNA is different as compared to A-
DNA
 Evidence for base stacking
CD/ORD of proteins
 e.g:- L-lysine
 Positive CD at 217nm
 Negative CD at 197nm
 Random coil :- ph 5.2
 Alpha helical :- ph 11.0
QUALITATIVE ANALYSIS
 Measuring by the specific rotation
 α =100 α /lc

 m =M α /100
Enzyme co-factor evaluation
 Teng et.al carried out the spectrum of
enzyme p-hydroxybenzoate
CD as finger printing
 It acts as finger printing technique for
trace amount of carbohydrate meterial
 E.g. dermatan sulphate
 heparitin sulphate
 E.g :- pectin in the presence of citric
acid& sucrose
CD of B-DNA & A-DNA
BASE STACKING
 ORD spectrum of Polycytidylic acid
shows at 292nm is 35,160deg/M/cm
 HELICITY DETERMINED BY
 Formaldehyde
 Ethylene glycol
DIFFERENCE B/W ORD &
CD
ORD CD
PLANE POLARIZED LIGHT
DISPERSIVE PHENOMENA
PLANE POLARIZED IS USED &
IS NOT CONVERTED TO
ELLIPTICAL LUGHT
GRAPHS ARE OBTAINED BY
SPECIFIC ROTATION V/S
WAVELENGTH
CIRCULARLY POLARIZED
LUGHT
ABSORPTIVE PHENOMENA
CIRCULAR POLARIZED LIGHT IS
USED & IS CONVERTED TO
ELLIPICITY
GRAPHS ARE OBTAINED
MOLAR ELLIPICITY V/S WAVE
LENGTH
INSTRUMENTATION OF
ORD
 Light source
 Monochromator
 Polarizer
 Analyzer
 Sample tube
 photomultiplier
INSTRUMENTATION OF CD
 Light source
 Polarizer
 Sample tube
 Photomultiplier
 Recorder
CD SPECTROSCOPE
Optical rotatory dispersion &                    circular dichorism
CD/ORD
 Simple and quick experiments
 No extensive preparation
 Relatively low concentrations/amounts
of sample
 Microsecond time resolution
 Any size of macromolecule
 Better resolution
 Better sensitivity
 Easier to assign
REFERENCE
 WWW. Science direct.com
 WWW.notes .com
 Principles & techniques by Avinash
Upadhyay
 WWW.scribd.com
 THANK YOU

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Optical rotatory dispersion & circular dichorism

  • 1. Optical Rotatory Dispersion & Circular Dichorism  Presented by:-  Sukhjeet kaur  M.Sc. (H) biotech(2)  130181106
  • 2. CONTENT  ORD  PRINCIPLE  FUNDAMENTAL OF ORD  CD  COTTON EFFECT  ORD CURVES  APPLICATION  DIFFERENCE B/W ORD & CD  CONCLUSION
  • 3. ORD  ORD is technique related to the optical activity  Rate of change of specific rotation with change in wave length  Optical rotation caused by compound changed with wavelength of light was first denoted by Biot in 1817
  • 4. FUNDAMENTAL OF ORD  Plane polarized light  Optical activity  Specific rotation  Circular birefringence  Optical rotation  Elliphicity
  • 5. Plane polarized light  Light from ordinary lamp consists of waves vibrating in many different planes  It is passed through Polaroid lens it is found to be vibrate in one plane is said to be plane polarized light
  • 7. OPTICAL ACTIVITY  The compounds which are having the ability to rotate the plane of polarized light are called optical active compound  This property of compounds is called optical activity  Measured by Polarimeter  Dextrorotatory  Levorotatory
  • 9. SPECIFIC ROTATION  The specific rotation ([α]) is an intensive property of a chemical compound, defined as the change in orientation of the plane of linearly polarized light as this light passes through a sample with a path length of 1 decimeter and a sample concentration of 1 gram per 1 millilitre  It is denoted by [α]
  • 10. CIRCULAR BIREFRINGENCE  Birefringence is the optical property of a material having a refractive index that depends on the polarization and propagation direction of light  α= (nL-nR)π/ α
  • 11. CIRCULAR DICHORISM Some material posses special properties of absorption of the light circularly light to different extent than the right circulary polarized light Chiral or asymmetric compounds produce CD spectrum
  • 12. COTTON EFFECT This phenomenon was discovered in 1895 by the French physicist Aimé Cotton Combination of circular dichorism & circular birefringence
  • 13. ORD CURVES  Plain curves  Anomalous curves  single cotton effect curves  multiple cotton effect curve
  • 14. PLAIN CURVES  Normal smooth curves  Do not cross the zero rotation line  e.g.:- alcohols hydrocarbons
  • 15. ANOMALOUS CURVES  Shows no. of extreme peaks & troughs
  • 16. SCEC & MCEC  Two or more peaks & trough  e.g. ketosteroids  camphor
  • 17. APPLICATION  CD/ORD of protein  Qualitative analysis of molecules  Enzyme co-factor evaluation  CD/ORD of proteins  CD as finger printing  CD/ORD of nucleic acids  CD of B-DNA is different as compared to A- DNA  Evidence for base stacking
  • 18. CD/ORD of proteins  e.g:- L-lysine  Positive CD at 217nm  Negative CD at 197nm  Random coil :- ph 5.2  Alpha helical :- ph 11.0
  • 19. QUALITATIVE ANALYSIS  Measuring by the specific rotation  α =100 α /lc   m =M α /100
  • 20. Enzyme co-factor evaluation  Teng et.al carried out the spectrum of enzyme p-hydroxybenzoate
  • 21. CD as finger printing  It acts as finger printing technique for trace amount of carbohydrate meterial  E.g. dermatan sulphate  heparitin sulphate  E.g :- pectin in the presence of citric acid& sucrose
  • 22. CD of B-DNA & A-DNA
  • 23. BASE STACKING  ORD spectrum of Polycytidylic acid shows at 292nm is 35,160deg/M/cm  HELICITY DETERMINED BY  Formaldehyde  Ethylene glycol
  • 24. DIFFERENCE B/W ORD & CD ORD CD PLANE POLARIZED LIGHT DISPERSIVE PHENOMENA PLANE POLARIZED IS USED & IS NOT CONVERTED TO ELLIPTICAL LUGHT GRAPHS ARE OBTAINED BY SPECIFIC ROTATION V/S WAVELENGTH CIRCULARLY POLARIZED LUGHT ABSORPTIVE PHENOMENA CIRCULAR POLARIZED LIGHT IS USED & IS CONVERTED TO ELLIPICITY GRAPHS ARE OBTAINED MOLAR ELLIPICITY V/S WAVE LENGTH
  • 25. INSTRUMENTATION OF ORD  Light source  Monochromator  Polarizer  Analyzer  Sample tube  photomultiplier
  • 26. INSTRUMENTATION OF CD  Light source  Polarizer  Sample tube  Photomultiplier  Recorder
  • 29. CD/ORD  Simple and quick experiments  No extensive preparation  Relatively low concentrations/amounts of sample  Microsecond time resolution  Any size of macromolecule  Better resolution  Better sensitivity  Easier to assign
  • 30. REFERENCE  WWW. Science direct.com  WWW.notes .com  Principles & techniques by Avinash Upadhyay  WWW.scribd.com