Polarimetry

Definition
Polarimetry is the measurement and interpretation of the polarization of transverse waves to the
direction of energy transfer, most notably electromagnetic waves, such as radio or light waves.
Typically, polarimetry is done on electromagnetic waves that have traveled through or have been
reflected, refracted, or diffracted by some material in order to characterize that object.
A polarimeter is the basic scientific instrument used to make these measurements.
Polarimetry can be used to measure various optical properties of a material, including linear
birefringence, circular birefringence (also known as optical rotation or optical rotary dispersion),
linear dichroism, circular dichroism and scattering.
Commonly used terms
# Polarization is a property of certain types of waves that describes the orientation of their
oscillations. Electromagnetic waves, such as light, and gravitational waves exhibit polarization;
acoustic waves (sound waves) in a gas or liquid do not have polarization because the direction of
vibration and direction of propagation are the same.
# Oscillation is the repetitive variation, typically in time, of some measure about a central value
or between two or more different states. Familiar examples include a swinging pendulum and AC
power.
# A transverse wave is a moving wave that consists of oscillations occurring perpendicular (or
right angled
# Reflection is the change in direction of a wavefront at an interface between two different media
so that the wavefront returns into the medium from which it originated. Common examples include
the reflection of light, sound and water waves.
# Refraction is the change in direction of a wave due to a change in its speed. It is essentially a
surface phenomenon
# Diffraction refers to various phenomena which occur when a wave encounters an obstacle. The
term diffraction, comes from the Latin diffringere, 'to break into pieces', referring to light breaking
up into different directions.
# Birefringence, or double refraction, is the decomposition of a ray of light into two rays when
it passes through anisotropic materials, such as crystals of calcite or boron nitride.
# Anisotropy is the property of being directionally dependent, as opposed to isotropy, which
implies identical properties in all directions. It can be defined as a difference, when measured along
different axes, in a material's physical or mechanical properties (absorbance, refractive index,
conductivity, tensile strength, etc.)
An example of anisotropy is the light coming through a polarizer. An example of an anisotropic
material is wood, which is easier to split along its grain than against it.
# Interference is a phenomenon in which two waves superimpose to form a resultant wave of
greater or lower amplitude.
Measurement of optical rotation
Optically active samples exhibit circular birefringence. Circular birefringence causes rotation of
the polarization of plane polarized light as it passes through the sample.
In an ordinary light, the vibrations occur in all planes perpendicular to direction of propagation.
When it is allowed to pass through a Nicol prism then its vibrations in the direction of axis is kept
intact and vibrations in all other directions are cut off. The light emerging out of the prism is said
to be plane polarized because its vibration is in one direction.
If two Nicol prisms are placed with their polarization planes parallel to each other, then the light
rays emerging out of the first prism will enter the second prism. As a result, complete bright light
is observed. If the second prism is rotated by an angle of 90°, the light emerging from the first
prism is stopped by the second prism due to which complete dark or no light region is observed.
The first prism is usually called polarizer and the second prism is called analyzer.
A simple polarimeter consists of a long tube with flat glass ends, into which the sample is placed.
At each end of the tube is a Nicol prism or other polarizer. Light is passed through the tube, and
the prism at the other end, attached to an eye-piece, is rotated to measure the region of complete
brightness or half-dark half -bright region or complete dark region. The angle of rotation is then
read from a scale. The same phenomenon is observed after an angle of 180°.
The specific rotation of the sample may then be calculated by using following formula.
Temperature can affect the rotation of light, which should be accounted for in the calculations.

100
[ ] 
lc
where:
[α]λT = the specific rotation
T = the temperature
λ = the wavelength of light,
σ = the angle of rotation
l = the length of the polarimeter tube,
c = the concentration of solution
Polarimeter
A polarimeter is a scientific instrument used to measure the angle of rotation caused by passing
polarized light through an optically active substance.
Some chemical substances are optically active, and polarized (unidirectional) light will rotate
either to the left (counter-clockwise) or right (clockwise) when passed through these substances.
The amount by which the light is rotated is known as the angle of rotation.
Construction
The polarimeter is made up of two Nicol prisms (the polarizer and analyzer). The polarizer is fixed
and the analyzer can be rotated. The light waves may be considered to correspond to waves in the
string. The polarizer allows only those light waves which move in a single plane. This causes the
light to become plane polarized. When the analyzer is also placed in a similar position it allows
the light waves coming from the polarizer to pass through it. When it is rotated through the right
angle no waves can pass through the right angle and the field appears to be dark. If now a glass
tube containing an optically active solution is placed between the polarizer and analyzer the light
now rotates through the plane of polarization through a certain angle, the analyzer will have to be
rotated in same angle.
Operation
Polarimeters measure this by passing monochromatic light through the first of two polarizing
plates, creating a polarized beam. This first plate is known as the polarizer. This beam is then
rotated as it passes through the sample. The sample is usually prepared as a tube where the optically
active substance is dissolved in an optically inactive chemical such as distilled water, ethanol,
methanol. Some polarimeters can be fitted with tubes that allow for sample to flow through
continuously.
After passing through the sample, a second polarizer, known as the analyzer, rotates either via
manual rotation or automatic detection of the angle. When the analyzer is rotated to the proper
angle, the maximum amount of light will pass through and shine onto a detector.
Applications
Because many optically active chemicals such as sucrose, are stereoisomers, a polarimeter can be
used to identify which isomer is present in a sample – if it rotates polarized light to the left, it is a
levo-isomer, and to the right, a dextro-isomer. It can also be used to measure optical activity of
racemic mixtures.
i) Chemical industry
Many chemicals exhibit a specific rotation as a unique property which can be used to distinguish
it. Polarimeters can identify unknown samples based on this if other variables such as
concentration and length of sample cell length are controlled or at least known. This is used in the
chemical industry.
By the same token, if the specific rotation of a sample is already known, then the concentration
and/or purity of a solution containing it can be calculated.
Most automatic polarimeters make this calculation automatically, given input on variables from
the user.
ii) Food, beverage and pharmaceutical industries
Concentration and purity measurements are important to determine product or ingredient quality
in the food & beverage and pharmaceutical industries. Samples that display specific rotations that
can be calculated for purity with a polarimeter include:
Steroids, Diuretics, Antibiotics, Narcotics, Vitamins, Analgesics, Amino Acids, Essential Oils,
Polymers, Starches,
iii) Sugars
Polarimeters are used in the sugar industry for determining quality of both juice from sugar cane
and the refined sucrose. Often, the sugar refineries use a modified polarimeter with a flow cell
called a saccharimeter.
Polarization
Polarization is a property of certain types of waves that describes the polarization orientation of
their oscillations. Electromagnetic waves, such as light, and gravitational waves exhibit
polarization. Acoustic waves (sound waves) in a gas or liquid do not have polarization because the
direction of vibration and direction of propagation are the same.
By convention, the polarization of light is described by specifying the orientation of the wave's
electric field at a point in space over one period of the oscillation. When light travels in free space,
in most cases it propagates as a transverse wave — the polarization is perpendicular to the wave's
direction of travel. In this case, the electric field may be oriented in a single direction (linear
polarization), or it may rotate as the wave travels (circular or elliptical polarization). In the latter
cases, the oscillations can rotate either towards the right or towards the left in the direction of
travel.
Polarization is significant in areas of science and technology dealing with wave propagation, such
as optics, seismology, telecommunications and radar science. The polarization of light can be
measured with a polarimeter. A polarizer is a device that affects polarization.
Types of Polarization
i) Linear polarization
• The two orthogonal (perpendicular) components are in a same phase.
• The ratio of amplitude of the two components is constant, so the direction of the electric
vector (the vector sum of these two components) is constant.
• Since the tip of the vector traces out a single line in the plane, this special case is called
linear polarization.
 • The direction of this line depends on the relative amplitudes of the two components.

ii) Circular polarization

• The two orthogonal components have exactly the same amplitude and are exactly ninety
degrees out of phase.
• One component is zero when the other component is at maximum or minimum amplitude.
• In this case the electric vector traces out a circle in the plane, so this special case is called
circular polarization.
• The direction the field rotates in depends on which of the two-phase relationships exists.
These cases are called right-hand circular polarization (if the x component is 900 ahead of
the y component) and left-hand circular polarization, (if the x component is 900 behind the
y component).
iii) Elliptical polarization
• The two components are not in a same phase.
• They do not have the same amplitude or are not ninety degrees out of phase. However,
their amplitude ratio is constant.
• They are called elliptical polarization because the electric vector traces out an ellipse in
the plane (the polarization ellipse).
• This polarization can rotate either to the right or left after specifying the direction of the
wave’s electric field.

Elliptical polarized light (violet) is composed of

unequal contributions of right (blue) and left (red)
circular polarized light.

Circular dichroism
Circular dichroism (CD) is a form of spectroscopy that refers to the differential absorption of left
and right circularly polarized light. It is exhibited in the absorption bands of optically active chiral
molecules.
CD spectroscopy has a wide range of applications in many different fields such as-
• UV CD is used to investigate the secondary structure of proteins.
• UV/Vis CD is used to investigate charge-transfer transitions.
• Near-infrared CD is used to investigate geometric and electronic structure by probing metal
d→d transitions.
• Vibrational CD, which uses light from the infrared energy region, is used for structural
studies of small organic molecules, and most recently proteins and DNA.
Principle
Linearly polarized light is polarized in a certain direction, that is the magnitude of its electric field
vector oscillates only in one plane. In circularly polarized light the electric field vector has a
constant length, but rotates about its propagation direction. Hence, it forms a helix in space while
propagating. If this is a left-handed helix the light is referred to as left circularly polarized and vice
versa for a right-handed helix.
The electric field of a light beam causes a linear displacement of charge when interacting with a
molecule, whereas the magnetic field of it causes a circulation of charge. These two motions
combined result in a helical displacement when light is impinged on a molecule. Since circularly
polarized light itself is "chiral", it interacts differently with chiral molecules and one of the two
types of circularly polarized light are absorbed to different extents.
In a CD experiment, equal amounts of left and right circularly polarized light are radiated into a
(chiral) solution. One of the two types is absorbed more than the other one and this wavelength
dependent difference of absorption is measured, yielding the CD spectrum of the sample. Due to
the interaction with the molecule, the electric field vector of the light traces out an elliptical path
while propagating.
When circularly polarized light passes through an absorbing optically active medium, the speeds
between right and left polarizations differ (cL ≠ cR) as well as their wavelength (λL ≠ λR) and the
extent to which they are absorbed (εL ≠ εR). Circular dichroism is the difference Δε ≡ εL- εR.
Application to biological molecules
1) In general, this phenomenon will be exhibited in absorption bands of any optically active
molecule. As a consequence, circular dichroism is exhibited by biological molecules, because of
their dextrorotary and levorotary components.
2) The secondary structure will also impart a distinct CD to its respective molecules. Therefore,
the alpha helix of proteins and the double helix of nucleic acids have CD spectral signatures
representative of their structures. The capacity of CD to give a representative structural signature
makes it a powerful tool in modern biochemistry with applications that can be found in virtually
every field of study.
3) CD is closely related to the optical rotatory dispersion (ORD) technique, and is generally
considered to be more advanced. CD is measured in or near the absorption bands of the molecule
of interest, while ORD can be measured far from these bands. CD's advantage is apparent in the
data analysis. Structural elements are more clearly distinguished since their recorded bands do not
overlap extensively at particular wavelengths as they do in ORD. In principle these two spectral
measurements can be interconverted through an integral transform (Kramers–Kronig relation), if
all the absorptions are included in the measurements.
4) The far-UV (ultraviolet) CD spectrum of proteins can reveal important characteristics of their
secondary structure. CD spectra can be readily used to estimate the fraction of a molecule that is
in the alpha-helix conformation, the beta-sheet conformation, the beta-turn conformation, or some
other (e.g. random coil) conformation. These fractional assignments place important constraints
on the possible secondary conformations that the protein can be in.
5) CD cannot, in general, say where the alpha helices that are detected are located within the
molecule or even completely predict how many there are. Despite this, CD is a valuable tool,
especially for showing changes in conformation. It can, for instance, be used to study how the
secondary structure of a molecule changes as a function of temperature or of the concentration of
denaturing agents, e.g. Guanidinium chloride or urea. In this way it can reveal important
thermodynamic information about the molecule (such as the enthalpy and Gibbs free energy of
denaturation) that cannot otherwise be easily obtained. Anyone attempting to study a protein will
find CD a valuable tool for verifying that the protein is in its native conformation before
undertaking extensive and/or expensive experiments with it. Also, there are a number of other uses
for CD spectroscopy in protein chemistry not related to alpha-helix fraction estimation.
6) The near-UV CD spectrum (>250 nm) of proteins provides information on the tertiary
structure. The signals obtained in the 250–300 nm region are due to the absorption, dipole
orientation and the nature of the surrounding environment of the phenylalanine, tyrosine, cysteine
(or S-S disulfide bridges) and tryptophan amino acids. Unlike in far-UV CD, the near-UV CD
spectrum cannot be assigned to any particular 3D structure. Rather, near-UV CD spectra provide
structural information on the nature of the prosthetic groups in proteins, e.g., the heme groups in
hemoglobin and cytochrome c.
7) Visible CD spectroscopy is a very powerful technique to study metal–protein interactions and
can resolve individual d–d electronic transitions as separate bands. CD spectra in the visible light
region are only produced when a metal ion is in a chiral environment, thus, free metal ions in
solution are not detected. This has the advantage of only observing the protein-bound metal, so pH
dependence and stoichiometries are readily obtained. Optical activity in transition metal ion
complexes have been attributed to configurational, conformational and the vicinal effects.
Klewpatinond and Viles have produced a set of empirical rules for predicting the appearance of
visible CD spectra for Cu2+ and Ni2+ square-planar complexes involving histidine and main-chain
coordination.
8) CD gives less specific structural information than X-ray crystallography and protein NMR
spectroscopy, for example, which both give atomic resolution data. However, CD spectroscopy is
a quick method that does not require large amounts of proteins or extensive data processing. Thus,
CD can be used to survey a large number of solvent conditions, varying temperature, pH, salinity,
and the presence of various cofactors.
9) CD spectroscopy is usually used to study proteins in solution, and thus it complements methods
that study the solid state. This is also a limitation, in that many proteins are embedded in
membranes in their native state, and solutions containing membrane structures are often strongly
scattering. CD is sometimes measured in thin films.