Transcript Slide 1
4. Molecular many electron systems: electronic & nuclear movement
Molecular
orbital
Electronic
configuration
Electronic
states
UV/Vis-absorption spectrum
* : antibinding
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4. Molecular many electron systems: electronic & nuclear movement
Jablonski-Scheme
Excitation [10-15 s]
v=1
Rotational levels
J=4
v=0
Microwavespectroscopy
J=3
J=2
J=1
J=0
UV-VIS-spectroscopy
S4
S3
Internal conversion
[10-14 s]
Tn
S2
IR- & NIRspectroscopy
Intersystem crossing
v=4
v=3
Vibrational levels
S1
T1
Fluorescence Phosphorescence
[10-9 s]
[10-3 s]
v=1
v=0
2
S0
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5. UV-Vis-Absorption
5.1 Franck-Condon principle
Interpret electronic absorption spectra based on
||2 of the vibrational levels
electronic transitions (~10-16s)
are much faster than the vibrational
period (~10-13s)
of a given molecule
thus nuclear coordinates do not
change during transition
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5. UV-Vis-Absorption
5.2 Franck-Condon principle
= degree of redistribution of electron density during transition
= degree of similarity of nuclear configuration between vibrational
wavefunctions of initial and final states.
Transition probability is proportional to the square modulus of the overlap integral
between vibrational wavefunctions of the two electronic states
= Franck-Condon-Factor:
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5. UV-Vis-Absorption
5.1 Franck-Condon principle
|f
|f
|i
|i
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5. UV-Vis-Absorption
5.2 Molecular electronic transitions
Molecular electronic transitions:
valence electrons are excited from one energy level to a higher energy level.
Electrons residing in the HOMO of a sigma bond can get excited
to the LUMO of that bond:
σ → σ* transition.
Promotion of an electron from a π-bonding orbital to an antibonding π* orbital:
π → π* transition.
Auxochromes with free electron pairs denoted as n have their own transitions, as
do aromatic pi bond transitions.
The following molecular electronic transitions exist:
σ → σ* π → π* n → σ* n → π* aromatic π → aromatic π*
p,p*
(C=C, C=O)
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np*
(C=O, C=N, C=S)
ns*
(–Hal, -S-, -Se- etc.)
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5. UV-Vis-Absorption
5.2 Transition metal complexes
A biologically very important group of metal complex bonds are the porphyrin
pigments such as:
Hemoglobin (pigment of the blood, central ion Fe2+)
octahedron structure motive
Heme-group
The four ligand positions of the base of
the pyramid are occupied by the lone
electron pairs of nitrogen atoms of the
plane porphyrin ring system
The two corners of the pyramid are
occupied by specific amino acids
(histidine) and/or by an oxygen molecule
(hemoglobin)
Cytochromes of respiratory chain
Chlorophyll (green molecules in leaves, central atom Mg)
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5. UV-Vis-Absorption
5.2 Transition metal complexes
Cytochrome C:
Pyramid corners of heme unit are occupied by N-atom of a histidine residue and
S-atom of a mezhionine residue
Redox change of cytochromes predominatly occurs at the central iron atom
[(Fe2+) ↔ (Fe3+)]
a-Peaks
= sensitive for redox change
(analysis of mitochondria)
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5. UV-Vis-Absorption
5.2 Transition metal complexes
Hemoglobin (iron is always found as Fe2+)
Arterial oxygen-loaded blood
=
light red
Blood in veines free of oxygen
=
deep red
°
0,4 A
End-on coordination of O2
(Fe2+ / 75 pm / low spin)
Desoxy Hemoglobin
(Fe2+ / 92 pm / high spin)
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B
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5. UV-Vis-Absorption
5.3 Polarimetry & Optical rotatory dispersion & Circular dichroism
Fundamental terms:
Polarimetry, optical rotation, circular birefringence:
turning of the plane of linearly polarized light
Optically active molecules exhibit
different refractive indices for right nR and left nL polarized light nR ≠ nL
Optical rotatory dispersion (ORD):
Wavelength dependency of rotation
Allows determination of absolute configuration of chiral molecules
Circular dichroism:
linearly polarized light is transformed into elliptically polarized light upon traveling
through matter
Different absorption coefficients for left and right circular polarized light
(eR ≠ eL ).
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5. UV-Vis-Absorption
5.3 Polarimetry & Optical rotatory dispersion & Circular dichroism
Polarimetry
What happens if light interacts with chiral molecules?
Enantiomeric molecules interact differently with circular polarized light.
Polarizability a depends on direction of rotation of incoming circular polarized light
Optically active substances exhibit different refractive indices for right nR and
left nL polarized light nR ≠ nL
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5. UV-Vis-Absorption
5.3 Polarimetry & Optical rotatory dispersion & Circular dichroism
Polarimetry:
Linearly polarized light
Different refractive index for its left and right circular constituents
Relative phase shift between left and right
Vector addition yields again linear polarized light with rotated polarization plane
Ey
phase
shift
Ex
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5. UV-Vis-Absorption
5.3 Polarimetry & Optical rotatory dispersion & Circular dichroism
Polarimetry: Due to the different refractive indices a phase difference d = jL –jR
builds up in the active medium which is proportional to the path
length l.
When exiting the medium linear polarized light where the oscillation
plane is rotated by d/2 arises
It follows:
For a follows:
l
a ( nL nR )
pl
l
Na-D line
l = 589 nm
2-Butanol
a = 11.2° (Messwert)
T = 20°C
l = 1dm
589109 m
(nL nR ) 11.2 deg
3.66107
180 deg 0.1 m
Difference is rather small!
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5. UV-Vis-Absorption
5.3 Polarimetry & Optical rotatory dispersion & Circular dichroism
Polarimetry
The measured angle-of-rotation results in:
a in angular degree, length in
decimeter(!) and c in g ml-1.
Specific rotation
is a substance specific constant (dependent on temperature
and wavelength) and is a measure for the optical activity of this particular substance.
Molar rotation is defined as follows:
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5. UV-Vis-Absorption
5.3 Polarimetry & Optical rotatory dispersion & Circular dichroism
Optical rotatory dispersion(ORD)
ORD measures molar rotation [F] as function of the wavelength!
If the substance to be investigated has
no electronic absorption within the
investigated spectral region the following
ORD spectra are obtained
ORD-spectra of 17ß- and 17ahydroxy-5a-androstan
Reason:
refractive indices for left and right
polarized light change differently with
wavelength (rotatory dispersion is
proportional to refractive index
difference).
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5. UV-Vis-Absorption
5.3 Polarimetry & Optical rotatory dispersion & Circular dichroism
Optical rotatory dispersion(ORD)
Refractive indices for left and right polarized light exhibit anomalous dispersion in the
range of an absorption band
Cotton effect
Positiv
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negativ Cotton effect
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5. UV-Vis-Absorption
5.3 Polarimety & Optical rotatory dispersion & Circular dichroism
Optical rotatory dispersion(ORD)
ORD-Spektren von 5a-Spirostan und 5a Quantitative theoretical correlations between
Spirostan-3-on
molecular structure and ORD (Cotton effect)
are difficult to derive;
Empirical investigation are important:
ORD has been successfully applied for
constitution elucidation e.g. to position
carbonyl groups in complex optically active
molecules.
By comparing ORD curves for structurally
isomeric ketons (reference material needed!)
the keto group can be localized.
ORD curve of molecule (2) is a superposition of a
negative curve i.e. molecular skeleton without a
chromophore (background curve) and a positive
Cotton curve (C=O chromophore).
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5. UV-Vis-Absorption
5.3 Polarimetry & Optical rotatory dispersion & Circular dichroism
Circular Dichroism (CD)
Enantiomeric molecules exhibit besides different refractive indices for left and
right circular polarized light also different absorption coefficients:
Circular Dichroism
It follows:
left and right circular components
ORD : different retardation
CD also different absorption
Ey
Ex
Transmitted light is elliptically polarized.
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5. UV-Vis-Absorption
5.3 Polarimetry & Optical rotatory dispersion & Circular dichroism
Circular Dichroism (CD)
The ratio between short and the long elliptical axis is defined as tangent of
an angle , the so called ellipticity (tan = b/a):
a = ER + EL
b = ER - EL
The specific ellipticity is defined as:
[10-1 × deg × cm2 × g-1]
where 0bs is the experimentally determined
ellipticity.
The molar ellipticity is defined as:
[10 × deg × cm2 × mol-1]
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5. UV-Vis-Absorption
5.3 Polarimety & Optical rotatory dispersion & Circular dichroism
Circular Dichroism (CD)
Signal heights are displayed either as absorption difference De or as ellipticity [q].
Molar ellipticity and circular dichroism can be interconverted:
[q] [grad cm2 dmol-1]
Correlation between ORD and CD:
ORD is based on the different refractive indices of left and right circular
polarized light (nR ≠ nL )
CD results from the different absorption behavior for left and right circular
polarized light (eR ≠ eL)
Connection of both phenomena via Kronig-Kramer relationship:
This relation allows the calculation of an ORD value for a particular wavelength l
from the corresponding CD spectrum
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5. UV-Vis-Absorption
5.3 Polarimetry & Optical rotatory dispersion & Circular dichroism
Circular Dichroismus (CD)
Simple model:
For an electronic transition to be CD active the following must be true:
µe is the electronic transition dipole moment
(= linear displacement of electrons upon transition into an excited state)
µm is the magnetic transition moment
(= radial displacement of electrons upon excited state transition)
Scalar product
is characterized by a helical electron displacement.
Depending on the chirality of the helix preferably more right or left circular
polarized light will be absorbed, respectively.
Electronic transition
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Magnetic transition
Optical activity
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5. UV-Vis-Absorption
5.3 Polarimetry & Optical rotatory dispersion & Circular dichroism
Circular Dichroism (CD)
Application field:
b-sheet
a-helix
random coil
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5. UV-Vis-Absorption
5.3 Polarimetry & Optical rotatory dispersion & Circular dichroism
Circular Dichroism (CD)
Application field:
Typical reference CD spectra:
Poly-L-Lysine in different conformations:
a-Helix, b-sheet and random coil.
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Temperature
dependent CD
spectra of insuline:
For increasing
temperature the
molecule changes
form a-helix into the
denaturated random
coil form with ß-sheet
contributions.
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5. UV-Vis-Absorption
5.3 Polarimetry & Optical rotatory dispersion & Circular dichroism
Vibrational-Circular-Dichroism (VCD)
Vibrational transitions in the IR and NIR
v=1
v=0
VCD monitors difference in absorption between left and right circular
polarized light
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5. UV-Vis-Absorption
5.3 Polarimetry & Optical rotatory dispersion & Circular dichroism
Vibrational-Circular-Dichroism (VCD)
()-Mirtazapine
Determination of the absolute
configuration
Advantages VCD vs. CD
Electronic chromophore is
not necessary
VCD exhibits more
characterisitic bands
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6. Fluorescence Spectroscopy
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6. Basic concepts in fluorescence spectroscopy
Excitation [10-15 s]
v=1
Rotational levels
J=4
v=0
Microwavespectroscopy
J=3
J=2
J=1
J=0
UV-VIS-spectroscopy
S4
S3
Internal conversion
[10-14 s]
Tn
S2
IR- & NIRspectroscopy
Intersystem crossing
v=4
v=3
Vibrational levels
S1
T1
Fluorescence Phosphorescence
[10-9 s]
[10-3 s]
v=1
v=0
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S0
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6. Basic concepts in fluorescence spectroscopy
6.1 Stokes-Shift
= Stokes-Shift due to vibrational energy
relaxation within electronic excited state
Energy differences between vibrational states which determine vibronic band
intensities are very often the same for ground and electronic excited state
Emission spectrum = mirror image of absorption spectrum
Emission bands are shifted bathochromically i.e. to higher wavelengths
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