Polarimetry

Theory & Principle Instrumentation Applications Disadvantages

Theory
Recall - Isomerism of Molecules

Theory
Stereoisomerism of Molecules

Have the same molecular formula and the same structural formula. However, different 3-D arrangement in space making them non-superimposable.

E.g. Stereoisomers D- and L- glyceraldehyde are non-superimposable mirror image molecules and are called enantiomers

Theory

Chiral Carbon

Two enantiomers of glyceraldehyde

Theory
Optical Isomerism / Rotation

A property exhibited by any compound whose mirror images are non-superimposable Enantiomers interact with plain polarized light to rotate the plane of the light in opposite directions This interaction with polarized light is called optical activity Measuring angle of rotation () - the concentration of a substance in a solution can be determined.

Principle
A plane polarized light is that which essentially has its vibration in only one direction or one plane as shown below (C)

A - the light waves propagating in all directions or planes (multidirectional) B - the Nicol prism which cuts all the planes of light and allows the light to come out with vibration in only one plane or direction (unidirectional) C - the resultant light which is said to be a plane polarized light.

Instrumentation - Polarimeter
Monochromatic light (Sodium vapor lamp) Polarized filter (Analyzer) mounted on circular dial rotated to give maximum amount of transmitted light Sample tube

Polarized filter - polarizer

Unpolarized light

Plane-polarized light

Rotated planepolarized light

degree of rotation measured

Instrumentation - Polarimeter

Unpolarized light from the light source is first polarized and passes through a sample cell. An optically active substance in a sample tube, rotates the plane polarized light. The rotation is noticed through the analyzer as a change in intensity of illumination. To reach the same illumination as was without an optically active sample, the analyzer must be turned around for an angle. Readings are taken in degrees (angle) a or sugar degrees Z.

Instrumentation - Polarimeter
Conventional polarimeter

Digital polarimeter

Half-shadow polarimeter

The Relationship Between Molecular Structure and Optical Activity When enantiomers in a solution is placed in the polarimeter, the plane polarized light is rotated by;

One isomer always rotates light in a clockwise (+) direction (dextrorotatory isomer) The other isomer rotates the light in a anticlockwise (-) direction (levorotatory isomer)

Instrumentation Polarimeter

Magnitude of rotation depends upon:

The nature of the compound The length of the tube (cell or sample container) usually expressed in decimeters (dm) The wavelength of the light source employed; usually either sodium D line at 589.3 nm or mercury vapor lamp at 546.1 nm Temperature of sample Concentration of compound in grams per 100 ml

Applications Sugar Analysis

Principle

Sugars with optically active molecules are capable of rotating plane polarized light. The rotation is directly proportional to the number of the molecules in the light path.

Specific rotation of a sugar, []DT, is 100 the angle of rotation of plane polarized light () of wavelength D after passing through 1 dm of a 1g/100 mL solution of the sugar at temperature T.

[] T
D

observed x 100 = lxc

D T obs l c []DT

= wavelength of the light source (Na) = temperature oC, normally 20C = observed rotation in degree (specify solvent) = length of tube in decimeter = concentration in grams/100ml = specific rotation (from references)

Therefore, measurements (unknown) can be either;

Sugar concentration or Specific rotation of sugar Types of sugar

D-glucose D-fructose D-galactose L-arabinose D-mannose D-arabinose D-xylose Lactose Sucrose Maltose+ Invert sugar Dextrin

+52.7 - 92.4 +80.2 +104.5 +14.2 -105.0 +18.8 +55.4 +66.5 +130.4 -19.8 +195

Specific rotation of various carbohydrates at 20C

Applications Determination of Starch Content

Principle
1st, sample is treated with hot dilute HCl. After clarification and filtration, the optical rotation of the solution is measured. 2nd,sample is extracted with 40 % ethanol. After acidifying the filtrate with HCl, clarifying and filtering, the optical rotation is measured. The difference between the two measurements, multiplied by a known factor, gives the starch content of the sample.

Starch concentration 2000 (P P) = (%) 20 []D

P P
20

= =

[]D =

total optical in angle degrees optical rotation in angle degrees of the substances soluble in 40% ethanol specific optical rotation of pure starch. The numerical values conventionally accepted for factor are the following;

Disadvantages

Requires the samples only in solution form. The solution should be free of bubble or any free particle, otherwise the light path will be affected and hence the accuracy.

The sample holder should be thermostatted, since the optical activities of the substances vary with temperature.