CH-SEC-208

By
Dr. Naveen
 Introduction
Polarimetry
▪ Polarimetry is a type of qualitative and quantitative technique, used for
optically active compounds.
▪ Polarimetry is the measurement and analysis of the rotation of plane-
polarized light as it passes through optically active (chiral) substances.
▪ It is used to determine the concentration, purity, and identity of
substances, such as sugars, pharmaceutical compounds, and steroids,
using an instrument called a polarimeter.

Polarimetry is one of the important instrumental methods employed in analysis. This

measures the rotation of the polarized light as it passes through an optically active
compound. This technique involves the measurement of change in the direction of
vibration of polarized light when interact with an optically active compound. A
substance is said to be optically active if it rotates the plane of the polarized light.
 Introduction
Plane Polarized Light
According to wave theory of light, an ordinary ray light is considered to be vibrating in all
planes at right angle to the direction of propagation. If this ordinary ray of light is passed
through a Nicol prism, the emergent ray has its vibration only in one plane. This light
having wave motion in only one plane is known as Plane Polarized Light.
Passing this through a Nicol prism—an optical device made of Calcite/Iceland Spar
(CaCO3)-eliminates one ray via internal reflection, producing plane-polarized light, which
vibrates in only one plane.
 Introduction
Plane Polarized Light
▪ A plane polarized light consists of two components of fixed magnitude rotating in
opposite directions to one another: The right circulatory polarized light and The left
circulatory polarized light.
▪ Plane polarized light is the vector sum of these two components.

Right and left circularly polarized

light are forms of electromagnetic
waves where the electric field
vector rotates in a circle, tracing a
helix along the propagation
direction. RCP (Right-Circularly
Polarized) light rotates clockwise
and LCP (Left-Circularly Polarized)
light rotates counterclockwise when
viewed from the receiver's
perspective
 Introduction
Optical Activity
It is the property of a chemical substance to rotate the plane of polarization
of plane-polarized light.
▪ Measured using a polarimeter, this phenomenon distinguishes chiral
materials, with clockwise rotation called dextrorotatory (+) and
counterclockwise, levorotatory (-).
▪ It is vital for identifying isomers and determining concentration.

Example: Lactic acid

Dextro= right designated by ‘d’, (+), clockwise.

Levo= left designated by ‘l’, (-), counterclockwise.
 Introduction
Optical Activity
Key Importances of Optical Activity:

•Pharmaceutical Analysis: Many drugs are chiral, and optical activity is used
to identify the correct active enantiomer. Using the wrong isomer can lead to
ineffective or harmful drugs.

•Quality Control & Purity: It measures the purity of chiral compounds. It

determines whether a substance is a pure enantiomer or a racemic mixture.

•Structural Chemistry: It provides insights into the 3-D structure of molecules,

helping scientists understand the spatial arrangement of atoms.

•Industry Applications: Widely used in the food and beverage industry to

determine the concentration of sugar (saccharimetry).

•Materials Science: Optically active materials like quartz, tourmaline, and

calcite are essential in manufacturing optical instruments, such as polarizing
filters and waveplate
 Introduction
Optical Rotation
▪ The optical rotation is the angle through which the plane of polarization is
rotated when polarized light passes through a layer of liquid.
▪ Optical rotation is the phenomenon where chiral (optically active)
substances rotate the plane of polarization of linearly polarized light,
typically measured using a polarimeter. It is used to determine
concentration, purity, and identity in industries like pharmaceuticals and
sugar refining.

Difference Between Optical Activity and Optical Rotation

▪ Optical activity is the capability of a substance (like sugar or quartz) to

interact with polarized light due to chirality. Optical rotation is the specific
numerical value of the angle by which the light plane is rotated.
▪ Measurement: Optical activity is a qualitative property (the
substance is or is not active), whereas optical rotation is a quantitative value
measured in degrees.
 Introduction
Specific Rotation
Specific rotation ([α]) is a characteristic physical property of chiral compounds,
defined as the angle of optical rotation observed when plane-polarized light passes
through a solution with a concentration of 1 g/ml at a path length of 1 dm, usually at
25 oC. It standardizes measurements for concentration and path length, allowing for
consistent comparison.

[α]D 25 = +3.12o
• means D line of a sodium lamp
(λ=589.6nm) is used for the light at
a temperature of 25oC, and that a
sample containing 1.00g/ml of the
optically active substance, in a 1-
dm tube, produces a rotation of
3.12o in a clockwise direction
 Introduction
Key Factors Affecting Specific Rotation

•Temperature (T): Specific rotation is temperature-dependent, as changing the

temperature can alter the density or the conformational equilibrium of the molecules.

Wavelength: The rotation changes with the wavelength of light used, a phenomenon
known as optical rotatory dispersion.

•Solvent: The solvent can affect the specific rotation by interacting with the solute
molecules, which can alter the compound's conformation or intermolecular interactions.

•Concentration: While observed rotation is directly proportional to

concentration, specific rotation is defined as the rotation per unit concentration, making
it generally constant for a given substance. However, at very high concentrations,
interactions between molecules might lead to slight variations.

•Structure/Nature of the Compound: The specific rotation is unique to each optically

active molecule and depends on its chiral environment.

•Path Length: Similar to concentration, the path length affects the observed rotation but
is normalized in the calculation of specific rotation.
 Working principle of a polarimeter

The working principle of a polarimeter lies in the interaction between polarized light
and an optically active substance. When polarized light passes through a sample,
the molecules of the substance rotate the plane of polarization. This rotation, known
as optical rotation, depends on the sample's nature, concentration, and the length of
the path the light travels through.
The polarimeter principle uses a light source, polarizer, sample tube, and analyzer.
The polarizer converts ordinary light into polarized light, which is then directed
through the sample. The analyzer measures the angle by which the light's plane has
been rotated.
This angle of rotation in polarimeter readings can be mathematically calculated
using the polarimeter equation to determine the substance's properties, such as its
concentration or optical activity
 Instrumentation

The Polarimeter is a device used to measure the effect of plane-

polarized light on optically active compounds. The components of
polarimeter are:

• A light source : usually a sodium lamp

• A polarizer: Nicol Prism

• A tube for holding sample in the light beam: sample cell

• An analyzer: Nicol prism aligned to intercept the linearly polarized ray

as it emerges from the sample solution.

• A scale: to measure the rotation of plane polarized light

Instrumentation
 Instrumentation

POLARIMETER: Working

• If no or optically inactive sample is present in the tube and the

instrument is reading zero (0o), the axes of plane polarized light and
the analyzer is exactly parallel.
• The observer will detect maximum amount (100 % transmittance) of
light passing through.
• If the sample is optically active the plane of PPL will be rotated as it
pass through the tube.
• In order to detect the maximum brightness of the light (ie. 100%
transmittance) observer will have to rotate the axis of the analyzer in
either clockwise or counterclockwise direction.
• If the analyzer is rotated in a clockwise direction, the rotation (α in
degree) is said to be positive (+), and such substance are c/a
dextrorotatory.
• If the rotation is counterclockwise, the α is –ve, and such substances
are c/a levorotatory
Instrumentation
Instrumentation
 Sample Preparation

Key Steps in Sample Preparation:

Sample Weighing: Accurately weigh the substance and dissolve it in a precise

volume of solvent using a volumetric flask to determine the concentration.

Solvent Selection: Use a solvent that dissolves the sample completely and is
optically inactive (typically water, methanol, or ethanol).

•Sample Filtration: Filter the solution if it is cloudy or contains suspended

particulates, as these interfere with light transmission.

•Tube Filling and Bubble Management: Fill the polarimeter cell/tube slowly to
avoid air bubbles. If bubbles are present, they should be moved to the bubble trap
(curved part) of the tube.

•Rinsing: Before filling, rinse the cell with the solvent, followed by a small amount
of the prepared sample solution
 Application
▪ Polarimetric method is a simple and accurate means for determination of structure
in microanalysis of expensive and non-duplicable samples.
▪ It is employed in quality control, process and research in the pharmaceutical,
chemical, essential oil, flavor and food industries.
▪ Isolating and identifying unknowns, crystallized from various solvents or separated
by HPLC.
▪ Evaluating and characterizing optically active compounds by measuring their
specific rotation and comparing this value with the theoretical values found in
literature.
▪ investigating kinetic reactions by measuring optical rotation as a function of time.
▪ Monitoring changes in concentration of an optically active component in a reaction
mixture, in enzymatic cleavage
.
▪ Analyzing molecular structure by plotting optical rotatory dispersion (ORD) curves
over a wide range of wavelengths.
▪ Distinguishing between optical isomer.
▪ To determine product purity by measuring specific rotation and optical rotation of:
Amino acids, Amino sugars, Analgesics, Antibiotics Cocaine, Dextrose, Diuretics,
Serums, Steroids, Tranquilizers, Vitamins etc.
▪ For raw materials inspection of: Camphors, Citric acid, Glyceric acid Gums
Lavender oil, Lemon oil Orange oil Spearmint oil.