Circular dichroism (CD)

Cindhy Jazmín Chamorro1

Neptun Code: BV8APY

1
Laboratory of Instrumental Analysis, Faculty of Science and Technology, University of
Debrecen.
Date of practice: October 26, 2022. Due date of report: October 28, 2022

1.Theoretical introduction
Optical activity is the capability of rotate the plane of linearly polarizes light or
transform from a linear polarization to an elliptical polarization light. This is mainly
caused by the arrangement and asymmetry of the molecular structure (due to their
chirality), due to the interaction between the light and matter. This interaction promotes
the change of the properties of the light such as the intensity, polarization, speed and
wavelength. In this case the most representative are the absorption and refraction.
Concerning the absorption process, the intensity and amplitude decrease because the
absorbance of part of the light. It is useful to get the concentration of a solution using
the Bouguer-Lambert-Beer law:

A=ε∗C∗l (Eq. 1)
Where:
l = Length of the cell
C = Concentration of the sample
ε = Molar extinction coefficient
A=Absorbance is defined as

( Iin)
A=log (Eq. 2)
( Iout)

In some cases, the matter does not absorb the light, so it slows down as it passes through
the body. Therefore, a difference in the refractive index is achieved. The refractive
index is described by:

Co
μ= (Eq. 3)
C
Where:
Co = Light velocity in the vacuum
C = Velocity in the matter
The optical activity is governed by two phenomena which manifest during the optical
activity:
1.1 Circular birefringence
It is an optical property of certain types of matter which consist of unfolding the light
into two polarized rays of light. Due to the enantiomers, the light acts in different ways.
On the one hand, the ray deviates of the right (rcpl) and the other to the left (lcpl), in
consequence some properties of the light change such as the velocity (decreases to the
initial one). In this case, one of the enantiomers reduces the speed more than the other
because of the excess of it, which means the refractive index will be different. The
difference between the lcpl and rcpl vector results in the rotation of the plane of the
resultant plane-polarized light with α (optical rotation) degree with respect to the
incident light phase:

( nl−nr )∗1800∗l
α= (Eq. 4)
φo

Where:
nl = Refractive index of lcpl
nr = rcpl refractive index
l = cell length
φo = speed of the light in the vacuum
The α value is concentration dependent, for that reason its more appropriate to use the
specific rotation [α ] or molar rotation [Ω]:

α
[α ] = (Eq. 5)
l∗C

[ α ]∗M
[Ω] (Eq. 6)
100

Where:
l = length measured
C= Concentration
M= Molecular weight
The molar rotation can be used to obtain the optical rotary dispersion (ORD) by plotting
it as a function of wave. When the ORD curve has an inflection point, a minimum a
maximum is known as cotton effect (CE).
1.2 Circular dichroism
The circular dichroism is an anomaly in the ORD spectrum caused by the absorbance of
the light into the active substance, thus the light passes through on absorbing optically
active medium and the velocity, wavelength and the degree of absorption between the
right and the left polarized differ.
It is defined as:

Io Io
∆ A= Al−Ar= − =∆ ε∗C∗L (Eq. 7)
Il Ir
Where:
C= Concentration
L= Cell length
Io= Intensity of incidence light
Il= Intensity of lcpl and Ir after passing the sample.
∆ ε = εl−εr
The transformation of the linearly polarized into elliptically polarized light is quantified
by the ellipticity (¥). If εl> εr means that it is greater than zero. In practice the value of
ellipticity commonly is measured in degree.
To obtain the CD spectrum it is necessary to measure ∆ A or ellipticity as a function of
the wavelength that is converted to ∆ ε by C or L during the processing.
The CD spectroscopy can be used to determine absolute configuration or absolute
conformation by these methods:
 Semiempirical rules
 Exciton coupled circular dichroism (ECCD)
 Computation of the rotary strength values and the CD spectra.

1.3 Circular Dichroism Spectroscopy

The circular dichroism spectroscopy is a type of spectroscopy to analyze the secondary
structure of proteins, to investigate the charge-transfer transitions, the geometric and
electronic structure, and used for structural studies of small organic molecules. The CD
spectropolarimeter is the instrument used for measures the samples. The CD
measurements could be performed in de UV-VIS and IR range. Also, the samples could
be measured in liquid and solid phase. The solid phase CD is very similar to IR
samples, it is used KCl or KBr. In this phase, it is necessary to obtain the
conformational analysis by X-ray.

2 Practical task
Plot the measured and calculated spectra of the given derivative, determine the absolute
configuration. (The 2D structure indicates the calculated enantiomer.)
The X-ray structure was available for this natural product including two solid-state
conformers. This clearly gave the relative configuration, but not the absolute one.
We measured the solid-state and the solution CDs of the substance and calculated for
the two conformers at the PBE0/TZVP level.
Plot the 3 spectra with Excel (or any other spreadsheet program) in one graph. For
scaling, the solid-state and the calculated spectra must be scaled to the measured one, if
necessary. (Indicate the degree of scaling.) The calculated CD has to be shifted to the
solid-state one.

3 Results and discussion

The results of the solid and solution phases provided by the spectrophotometer were
plotted. Additionally, the X-ray derived computed ECD was plotted too. Concerning the
solid-state, it was scaled up multiplying by 8 to adjust to the graphic. Moreover, it was
necessary to shift the compute curve, in order to fit the CD spectrum.