Refractometry

Refractometry is a technique that measures how light is refracted when it passes

through a given substance, in this case, an unknown compound.

The amount by which the light is refracted determines the refractive index. Refractive
index can be used to identify an unknown liquid compound, or it can be used as a
means of measuring the purity of a liquid compound by comparing it to literature
values.

The closer the refractive index is to the literature values, the purer the sample.
Refractive index is defined as the ratio of the velocity of light in air to the velocity of
light in themedium being measured:
nD = [V air]/(V liquid)

Refractive index is temperature dependent. To adjust for temperatures greater than

20°C, please use the following formula:
nD20 = nDT + 0.00045(T - 20oC)

Apparatus
This is a view of a refractometer.

Procedure

1. Lift prism and clean both top and bottom surfaces with 95% ethanol.
2. Once both surfaces are dry, place just enough of your neat (undiluted) sample
onto the bottom prism until the entire surface is covered using a disposable
pipette.
3. Close the prism and raise the light as far as it will go.
4. Turn the knob on the front of the instrument to n D. You will see that the light
turns on.
5. Look into the eyepiece. Turn the knob on the right side of the instrument until
you see a black line.

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 6. Use the dial directly under the eyepiece to focus the line so it is a clear as you
can get it.
7. Again, using the knob on the right side of the instrument, adjust the line so
that it intersects with the cross hairs you see in the eyepiece.
8. Press the READ button and record the value that shows on the digital readout.
This is nDT.
9. Press the TEMP button and record the value (T) that shows on the digital
readout.
10. Use the equation above to calculate nD20.
11. Clean both top and bottom surfaces of the prism with 95% ethanol. Place a
clean folded piece of wipe on the bottom prism and close the prism lid.

Polarimetry
(http://academics.wellesley.edu/Chemistry/chem211lab/Orgo_Lab_Manual/
LabManual/index.html,
http://rudolphresearch.com/products/polarimeters/polarimetry-definitions/)

Polarimetry measures the extent to which a substance interacts with plane polarized
light (light which consists of waves that vibrate only in one plane);

whether it rotates plane polarized light to the left, to the right, or not at all. If the
substance rotates plane polarized light to the left or to the right, it is called optically
active.

To be optically active, a compound must have a chiral center.

A chiral center is a carbon that has 4 different groups attached to it.

Depending on the orientation of these four different groups about the chiral carbon,
the compound may rotate plane polarized light to the left or to the right.

If a compound does not have a chiral center, it will not rotate light at all.

The number of degrees and the direction of rotation are measured to give the observed
rotation.

The observed rotation must be corrected for the length of the cell used and the
solution concentration.

Comparing the corrected observed rotation to literature values can aid in the
identification of an unknown compound.

Polarimetry measures the rotation of polarized light as it passes through an optically

active fluid.
The measured rotation can be used to calculate the value of solution concentrations;
especially substances such as sugars, peptides and volatile oils.
A polarimeter consists of a polarized light source, an analyzer, a graduated circle to
measure the rotation angle, and sample tubes.
The polarized light passes through the sample tube and exhibits angular rotation to the
left (-) or right (+). On the side opposite the polarizer is the analyzer.

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Using optics, visual fields are manually adjusted by the user to measure the optical
rotation angle.

Schematic diagram

Polarimeters offer high accuracies where precision is critical in determining the

concentration of samples. Cole-Parmer offers manual polarimeters where you look
through a viewing scope to read values on a vernier scale, and semiautomatic
polarimeters that have a digital display.
Polarimeters can measure in angle of rotation (α), International Sugar Scale (°Z), or
both.

Automated Polarimeter  Manual Polarimeter

Polarimetry Fundamentals

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Polarimetry is a sensitive, non-destructive technique for measuring the optical activity
exhibited by inorganic and organic compounds.
A compound is considered to be optically active if linearly polarized light is rotated
when passing through it.
The amount of optical rotation is determined by the molecular structure and
concentration of chiral molecules in the substance.
Each optically active substance has its own specific rotation as defined in Biots law:

Historically, polarimetry was performed using an instrument where the extent of

optical rotation is estimated by visual matching of the intensity of split fields.
For this reason, the D-line of the sodium lamp at the visible wavelength of 589 nm
was most often employed.

 
It is not common practice to use other light sources, such as xenon or tungsten
halogen, with appropriate filters, because these may offer.

Advantages:
cost,
long life,
broad wavelength emission range over traditional light sources
 
 

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Calibration of Polarimeter
 
Method:-Normal sucrose solution prepared from exactly 26.00g of sucrose dissolved
in pure water to 100 cm3. Normal Sucrose Solution was used to calibrate and
standardize polarimetric
methods and instruments.
 
Main Reasons to use sucrose solutions:
1.     Very easily prepared
2.     Has a fixed rotation value which can calibrated with 10%, 20% and 30% sugar
value, and remains constant at those  concentrations for a little time only, up to that we
can have a very sharp reading calibration for pure sugar.
3.    When a polarimeter is used for sucrose quality detection, calibration with pure
sucrose is the best.
4.    Its shows almost no muta- rotation and lastly, it is cheap to use in order to get the
calibration done.

 
Applications for Polarimeter:-
 Sugar Industry
 Pharmaceutical Industry
 Chemical Industry
 Flavours, Fragrances and Essential Oils
 Starch
 Food and Drink
 Agriculture
 Amino acids
 Monosodium Glutamate

What are the advantages of automatic laboratory polarimeters?

Polarimetry is a powerful technique for the analysis of optically-active fluids such as
sugars, lactic acid, tartaric acid, and biologically-active substances.

The method provides valuable information on the chemical structure, chirality, and
concentration of a sample by measuring the angle through which a ray of polarised
light is diverted.

In a traditional manual polarimeter, this requires a skilled operator and takes a

variable length of time.

A modern automatic polarimeter, on the other hand, makes the process much quicker
– an accurate reading is provided in just one second.

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Operator-induced errors are eliminated and measurement consistency is greatly
improved – an important consideration for regulated laboratories.

Touchscreen controls and a clear digital readout make the analysis fast, clean, and
reliable.

Automatic polarimetry is compatible with Good Laboratory Practice (GLP) and US

FDA 21 CFR part 11.

The increased speed means that lab productivity is greatly improved, and the
investment in an automated instrument pays real dividends.

Manual polarimetry still has its place, for low-throughput labs and for training, but for
many modern industries such as pharmaceutical, chemical, sugar, and food
processing, an automatic polarimeter is the better choice.