OPTICAL ROTATION
PRESENTED BY
ANIKET KALE
23MSPA006
PA-620
CONTENTS
• OPTICAL ROTATION
• SPECIFIC OPTICAL ROTATION
• CALIBRATIONS METHODS
• PRINCIPLE OF POLARIMETER
• APPLICATIONS OF OPTICAL ROTATION
• REFERENCES
OPTICAL ROTATION
• Optical rotation 'a' is the property shown by certain substances of
rotating the plane of polarisation of polarised light.
• Such substances are said to be optically active in the sense that they
cause incident polarised light to emerge in a plane forming a
measurable angle with a plane of the incident light.
• Where this effect is large enough for measurement, it may serve as
the basis for identifying or assaying a substance.
• The optical rotation of a substance is the angle through which the
plane of polarisation is rotated when polarised light passes through
the substance, if liquid, or a solution of the substance.
• Substances are described as dextro-rotatory or levo-rotatory
according to whether [a] the plane of polarisation is rotated clockwise
or anticlockwise, respectively as determined by viewing towards the
light source.
• Dextro-rotation is designated (+) and levo-rotation is designated
(-).
• The optical rotation unless otherwise specified, is measured at the
wavelength of the D line of sodium (λ= 589.3 nm) at 25°, on a layer
1 dm length. It is expressed in degrees..
CAUSE OF OPTICAL ROTATION
Major cause of the optical rotation:
CHIRALITY
Carbon atom with four different groups attached to it.
ASYMMETRY
Molecule not divided equally in a plane into two equal halves.
FACTORS AFFECT THE OPTICAL ROTATION
The angle of rotation may have influenced by:
1. Concentration of the sample, but SOR will not change, since concentration is
consider in the calculation.
2. Wavelength of light passing through the sample. Angle of rotation and wavelength
tend to be inversely proportional.
3. Temperature of the sample (generally the two are directly proportional).
4. Path length of the sample cell.
5. Solvent used for solution preparation.
6. Operation: Filling conditions (bubbles, temperature and concentration gradients).
SPECIFIC OPTICAL ROTATION
• The specific optical rotation of a liquid substance is the angle of
rotation of the plane of polarisation at the wavelength of the D line of
sodium (λ = 589.3 nm) measured at 25°, unless otherwise specified,
calculated with reference to a 1-dm thick layer of the liquid, and
divided by the specific gravity at 25°.
• The specific optical rotation of a solid substance is the angle of
rotation of the plane of polarisation at the wavelength of the D line of
sodium (λ= 589.3 nm) measured at 25°, unless otherwise specified,
calculated with reference to a 1-dm thick layer of a solution
containing 1 g of the substance per 100 ml.
• The specific optical rotation of a solid is always expressed with
reference to a given solvent and concentration.
CALIBRATION
• The apparatus may be checked by using a solution of previously dried
sucrose and measuring the optical rotation in a 2-dm tube at 25° and
using the concentrations indicated in the table.
METHOD FOR SOLIDS
• Weigh accurately a suitable quantity of the substance under
examination to obtain the solution of the strength specified in the
individual monograph and transfer to a volumetric flask by means of
water or other solvent, if specified.
• If a solvent is used, reserve a portion of it for the blank determination.
Unless otherwise specified, adjust the contents of the flask to 25º by
suspending the flask in a constant-temperature bath.
• Make up the volume with the solvent at 25° and mix well. Transfer
the solution to the polarimeter tube within 30 minutes from the time
the substance was dissolved and during this time interval maintain the
solution at 25°.
• Determine the zero point of the polarimeter and then make five
readings of the observed rotation of the test solution at 25°.
• Take an equal number of readings in the same tube with the solvent in
place of the test solution.
• The zero correction is the average of the blank readings, and is
subtracted from the average observed rotation if the two figures are of the
same sign or added if they are opposite in sign to obtain the corrected
observed rotation.
METHODS FOR LIQUIDS
• Unless otherwise specified, adjust the temperature of the substance under
examination to 25°, transfer to a polarimeter tube and proceed as described
for solids, beginning at the words "Determine the zero point“.
POLARIMETER
PRINCIPLE
Un-polarized light from the light source is first polarized.
This polarized light passes through a sample cell.
If an optical active substance is in a sample tube, the
plane of the polarized light is rotated.
The rotation is noticed by looking through the analyzer
as a change in the intensity of illumination.
To reach the same illumination as was without an optical
active sample the analyzer must be turned around for an
angle.
Reading is taken in degree angle.
REQUIREMENT
A commercial instrument constructed for use with a sodium lamp and
capable of giving readings to the nearest 0.02° is suitable for most
purposes.
For certain applications, the use of a photoelectric polarimeter capable of
taking measurements at the specific wavelengths may be necessary.
The accuracy and precision of optical rotation measurements can be
increased if the following precautions are taken.
The instrument must be in a good condition. The optical elements must
be very clean and in exact alignment. The match point should be close
to the normal zero mark.
The light source should be properly aligned with respect to the optical
bench. It should be supplemented by a filtering system capable of
isolating the D line from sodium light.
Specific attention should be paid to temperature control of the solution
and of the polarimeter.
Differences between the initial readings or between observed and
corrected optical rotation, calculated as either specific optical rotation or
optical rotation, should not be more than one-fourth of the range specified
in the monograph for the substance.
Polarimeter tubes should be filled in such a way as to avoid air bubbles.
Particular care is necessary for semi- micro or micro tubes.
For tubes with removable end-plates fitted with gaskets and caps,
tighten the end-plates only enough to ensure a leak- proof seal
between the end-plate and the body of the tube.
For substances with low rotatory power, the end-plates should be
loosened and tightened again after each reading, in the measurement
of both the rotation and the zero point.
Liquids and solutions of solids must be clear.
INSTRUMENTATION
The Polarimeter typically consists of:
• A light source, for example a sodium discharge lamp, a (LED) light-emitting
diode (LED) or another light source capable of providing radiation at the
desired wavelength (589 nm unless otherwise prescribed in the monograph); if
the light source is polychromatic, a means of isolating the required wavelength
is necessary. e.g. an optical filter
• A polariser and an analyser
• A sample cell with a path length of 1.00 dm, unless otherwise specified in the
monograph
• A detection system to measure the angle of optical rotation, which must be
capable of giving readings to at least the nearest 0.01°, unless otherwise
specified in the monograph.
• A temperature control system that indicates the temperature
with a readability of 0.1 °C; it may be embedded in the
polarimeter (e.g. a Peltier system) or be an external unit (e.g. a
cycle-cryostat), and must be able to maintain the temperature of
the liquid to within ± 0.5 °C of that prescribed.
EQUIPMENT PERFORMANCE
• The accuracy of the scale is checked near the value to be
measured or over an appropriate range, usually by means of
certified quartz plates.
• Other certified reference materials may also be suitable.
(e.g. sucrose solutions).
• Optical rotation measurements may be used to quantify the
amount of an enantiomer or the ratio of enantiomers present in
a sample.
• For that purpose, the linearity must be checked, for example
using sucrose solutions.
APPLICATIONS OF OPTICAL ROTATION
Optical activity is only parameter available for distinguishing between D and
L isomeric forms.(Qualitative)
This is also used to plot optical rotatory dispersion curves for a various range
of wavelengths this helps in analysing molecular structure.
Optical rotation is a function of time and it is used to determine kinetic
reactions.
If the specific rotation of a sample is known its concentration in the solution
can be estimated and Vice-versa.(Quantitative)
The technique may be extended to the determination of optical substances in
the presence of optically inactive species.
REFERENCES
• INDIAN PHARMACOPOIEA
• BRITISH PHARMACOPOIEA
• EUROPEAN PHARMACOPOEIA
THANK YOU