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Title:To carry out assay of glucose in oral rehydration salts by polarimeter.

Theory:
Certain substances, in liquid or in solution, are capable of rotating the plane of polarized light(vibrating in only one plane) ,either to the right (dexter) or to the left(laevus) are called optically active or chiral compounds. And the instrument which is mean to generate or isolate plane polarized light and to detect, measure the extent of rotation of the plane of polarized light is called polarimeter. 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 polarization is rotated when polarized light passes through the substance ,if liquid, or a solution of the substance. Substances are described as dextrorotatory [(+),rotation towards right side)] or laevo-rotatoty[(-)rotation towards left side)]. The optical rotation or the angle of rotation() is dependent upon the nature of the substance, length of the column of solution through which the light passes, the temperature, concentration of the solution and the wavelength of the light used. The optical rotation is measured at 20C or at 25C and with necessarily using monochromatic light source. The rotation() , is thus given by: = []t.l. m/v Where, l is the length of the column of the solution in decimeter(dm) through which the light passes, m/v is weight in g of the substance dissolved in a volume in ml. []t is a constant and called specific optical rotation. It depends upon the nature of substance, temperature and wavelength of light. It is generally expressed at 20C or at 25C using sodium D-line(589.3nm), hence the relation becomes, []25D= .v/l.m The constant is expressed in degree of rotation per decimeter per gram, ml(deg dm-1,g-1ml) but when l= 1dm, v=1ml, m=1g, then optical rotation becomes equal to Sp. optical rotation. []25D= , If c is g of substance is dissolved in 100ml of water or solvent, the relation becomes, []25D= 100./l.c

For a liquid substance the relation becomes, []25D= 100./l.d25,where d25 is the specific gravity of the liquid sample at 25C. Instrumentation (Visual polarimetry):

In the visual polarimetry , the light source is usually a sodium vapour lamp . A beam of natural light consists of a vast number of rays of linearly polarized light vibrating perpendicular to the direction of propagation. The planes of electric field vectors are randomly oriented, hence called unpolarized. The randomly oriented planes of oscillation of natural light can be reduced to two planes perpendicular to each other, hence, the unpolarized light can be resolved into two components which can be represented by the symbol . The light from the sodium vapour lamp is made parallel by lens L1. Island spar(Nicol prism,P1) possesses different refractive indexes for the mutually perpendicular components (,) of unpolarised light. Hence, P1 (Polarizer) resolves the natural light into two rays of linearly polarized light. One is allowed to pass through while another is totally reflected(a) back into the crystal. In the absence of optically active material in the sample tube and when the planes of the prisms are parallel, plane polarized light passes through the P3(Analyzer) as in (a). If P3 is rotated about the axis of the instrument so that the principal planes of P1 and P3 are mutually at right angles(as in b); as a result no light is transmitted by P3. This condition of maximum extinction is achieved by crossed Nicols. This condition of total extinction and illumination is achieved alternately when the P3 is rotated through 90, ,180 , 270 and 360. If now an optically active sample is placed in the tube, light will once more emerge from P3. Extinction can be restored by rotating P3 through an angle(), measured on a calibrated major and vernier scale. It is difficult to determine precisely at which angular position of P3 the illumination of the field at a minimum because the transmitted intensity change per unit angular rotation very small near the extinction point(which our eyes may not differentiate precisely). Hence, A third prism P2 (or a half wave quartz plate) is therefore inserted to cover the half field. This causes two planepolarized rays to pass through the tube, differing in phase by half a wavelength.

If the analyzing prism is crossed with the unobstructed part of the field the part is darkened, as in A, but some light is present in the obstructed half. Rotation of the analyzer in the correct direction gives a field as in B, in which both beams are almost completely extinguished or equally illuminated, and both area of the field are of equal intensity(end-point). Further rotation gives C, in which light from the obstructed half of the field is eliminated, but some rays from the unobstructed half pass through. When the analyzer has to be rotated to the right to get the endpoint, the substance is said to be dextro-rotatory and if it to be rotated to left then the substance is said to be laevo-rotatory.

Chemicals required:
1)ORS sachets, 2) Dil.Ammonia solution(10%).

Apparatus required:
1)Polarimeter, 2)Volumetric flasks(50ml), 3)Pipettes.

Procedure:
Weigh accurately about 8.0 g of powder, dissolve in 40 ml of water, add0.5 ml of dilute ammonia solution to increase the rate of mutarotation and dilute to 50 ml with water. Mix well, allow to stand for 30minutes, filter the solution if turbid. Switch sodium lamp on and allow for 10 minutes for full intensity illumination. Then, wasth 2-dm tube with water and fill it with water taking care to avoid the presence of bubbles in the tube. Screw the metal cap tightly. If air bubbles are found in the tube, trap them in the protruded part of the tube. Then place the tube in compartment. After this step, move the analyzer about the zero, as marked on the graduated scale, until the field of vision is equally illuminated. Approach this position three times from either side. This position is read on the scale with the help of a vernier. If the zero of the vernier does not lie exactly at the zero graduation of the main scale, all the readings must be corrected for the displacement of the zero. The angle by which the zero is displaced must be added to or subtracted from observed rotation of sample solution according the modes of rotation (dextro- or laevo-) . Then empty the tube and rinse it with sample solution and fill it. Then measure the optical rotation as in solvent(Three determinations from each side). The observed rotation in degrees multiplied by 0 .9411 and 1.0424 represents the weight in g of C6H12O6 and C6H12O6, H2O respectively as appropriate in the weight taken for the assay. Weight of ORS sample taken= 40ml water + 0.5ml Dil. ammonia Soln. 50ml with water.

Observation:
Solvent(Blank) Optical rotation measured from right side Optical rotation measured from left side. 1) 2) 3) 1) 2) Sample Optical rotation measured from right side Optical rotation measured from left side. 1) 2) 3) 1) 2)

3) Mean optical Rot. Blank Mean observed optical. Rot. Sp. Actual optical rot(Sp. Blank)

3)

Calculation:
Each sachet contains (glucose, anhydrous/monohydrate) = Degree of optical rotation X 0.9477 X Net.Wt.of contents in each sachet Concentration of sample(%w/v)

Limits: For anhydrous form (12.15-14.85)g Each sachet contains (glucose,monohydrate)

=

Degree of optical rotation X 1.0424 X Net.Wt.of contents in each sachet Concentration of sample (%w/v)

Limits: For monohydrate form (13.37-16.34)g

Result: Conclusion:

Performed By Name: Roll No.:

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