Introduction

Refraction happens when a light bends because it passes at an angle through a boundary between
isotropic media. When the angle of incidence in water reaches a certain critical value, the refracted ray
lies along the boundary, having an angle of refraction of 90-degrees. This is called the critical angle and
is the largest angle of incidence for which refraction can occur.

In a natural setting, there are two types of light that occurs. Unpolarized light is a light wave that vibrates
more than in one plane (candle light, sun light). These light waves are created by electric charges that
vibrates in variety of directions. On the other hand, polarized light is a light wave that vibrates in a single
plane. To change the unpolarized light wave to a polarized light wave, a polarization process occurs.

In chemistry, optical isomers are isomers which affects plane polarized light. Optically active substances
can rotate the plane of polarization either clockwise or counterclockwise. A substance that is an optical
isomer exists as two isomers known as enantiomers. Optically active substances synthetically produced
in a lab can exist as a 50/50 mixture of two enantiomers. This is called racemic mixture.

In the pharmaceutical setting, refractometry is widely used when the material being analyzed is known. It
is commonly used in calculating the concentration of dissolved substances in water. Polarimetry, in
pharmaceutical setting, can determine a product purity by its specific rotation and optical rotation. Some
common products are amino acids, dextrose, serums and vitamins.

In this experiment, an Abbe refractometer was used. This apparatus relies on the measurement of critical
angle. A sample will be placed in two prism for measuring and illuminating. Light enters sample from the
illuminating prism, gets refracted at critical angle at the bottom surface of measuring prism, and then the
telescope is used to measure position of the border between bright and light areas. Telescope reverts
the image, so the dark area is at the bottom, even if we expect it to be in the upper part of the field of
view.

In this refractometry experiment, dextrose standard solution was used. The theoretical refractive index of
dextrose at 20oC is 1.3477. In polarimetry experiment, dextrose, sucrose and inverted sucrose was used.
The theoretical optical rotation were +52.70, & +66.47

Methodology – Refractometry

For the preparation of standard dextrose solutions, 0.1 g, 0.5 g, and 1.0 g dextrose monohydrate were
dissolved in about 5.0 mL of distilled water in a 50.0 mL beaker. The standard solutions were then
transferred to separate 10.0 mL volumetric flasks and diluted with distilled water to mark.

Abbe Refractometer was used in the experiment. The prisms were cleaned with lens paper saturated
with 95% ethanol and was calibrated with distilled water prior to experiment. Two to three drops of
dextrose solution were placed in the refracting prism. The light source (fluorescent lamp) was adjusted to
be reflected through the prisms until the borderline between the light and dark halves of the field of view.
Compensator prisms were adjusted to obtain a sharp resolution of uncolored border line. Refractive
index was read directly from the graduated scale to the fourth decimal place. Three trials were done for
this experiment and the temperature at which each reading were carried out were noted. The refractive
index of an unknown dextrose solution was also tested in three trials

A plot of refractive indices of the standard dextrose solutions and molar concentration was then provided.
Methodology – POLARIMETRY

For the preparation of sucrose solution, 26 g of sucrose was dissolved in about 50 mL of distilled water
and was transferred to a 100.0 mL volumetric flask and was diluted with same solvent to mark. Two 25-
mL were then pipetted from the volumetric flask and was place on separate beakers. The first 25-mL
sample was used for the determination of optical rotation of sucrose. The second 25-mL sample was
subjected to 0.1 mL of concentrated hydrochloric acid and was heated gently in water bath for 30
minutes at 50oC. Fehling’s test was carried out until the sample produces a brick-red precipitate.

To determine to optical activity of sugar solutions, an amount of each solution was placed in a 1-dm
polarimetry tube. No bubbles were observed in the tube during the placement of samples. The tube was
cooled to 20-25oC. Until such time, the tube was placed in the sample compartment and closed. The
knob was rotated clockwise and counterclockwise until a half-shade end point is observed. The angular
rotation was determined from the Vernier scale. From these readings, the specific rotation was
calculated.

Preliminary Data
Weight of glucose: 10.08 g; volume: 100.00mL
Weight of sucrose: 26.00g; volume: 100.00mL
Invert Sugar; volume of aliquot: 25.00mL

Results
ANALYSIS OF DEXTROSE SOLUTION BY REFRACTOMETRY

Dextrose Weight of Molar Refractive Index (Temperature, 0C)

Solution Dextrose Concentration
Monohydrate Trial #1 Trial #2 Trial #3

Standard 0.1077g 0.05 M 1.3350 @ 27oC 1.3351 @ 27.3oC 1.3351 @ 27.4oC

Solution #1
Standard 0.5081g 0.26 M 1.3405 @ 27oC 1.3410 @ 27.3oC 1.3401 @ 27.4oC
Solution #2
Standard 1.0071 0.51 M 1.3475 @ 27oC 1.3355 @ 27.3oC 1.3471 @ 27.4oC
Solution #3
Unknown -- -- 1.3471 @ 27oC 1.3473 @ 27.3oC 1.3475 @ 27.4oC
Solution

ANALYSIS OF SUGAR SOLUTIONS BY POLARIMETRY

Invert Sugar
Fehling’s test: positive (brick red precipitate)

Solution Angular Rotation Specific Rotation

10% Dextrose 10.7 O 53.5 O
26% Sucrose 15 O 57.69 O
Invert Sugar 8.85O 34.04 O
 1.35

1.348

1.346
Refractive Index

1.344

1.342

1.34

1.338

1.336

1.334
0 0.5 1 1.5 2 2.5 3 3.5
Molecular Concentration

Chart #1: Refractive indices of standard solutions at 27.0oC (trial 1)

1.342

1.341

1.34
Refractometry

1.339

1.338

1.337

1.336

1.335

1.334
0 0.5 1 1.5 2 2.5 3 3.5
Molecular Concentration

Chart #2: Refractive indices of standard solutions at 27.3oC (trial 2)

 1.348

1.346

1.344
Refractive Index

1.342

1.34

1.338

1.336

1.334
0 0.5 1 1.5 2 2.5 3 3.5
Molecular Concentration

Chart # 3: Refractive Indices of standard solution at 27.4oC (trial 3)

Discussion
In refractometry, the theoretical refractive index of dextrose is 1.3477 at 20oC. In our experiment, due to
unseen circumstances, the temperature recorded by the Abbe refractometer is higher by +7 oC which
played a role in a change in refractive index of standard solutions. The recorded refractive indices which
averages at 27.3OC is 1.390. In theory, the refraction index decreases as the temperature increases. The
speed of light in the sample is slower because the light can be absorbed and reemitted by the atoms in
the sample. With an increase in temperature, the refractive indices are expected to be lower since the
density of the liquid sample is affected.
In polarimetry, the theoretical rotation of specific rotation of dextrose and sucrose is +52.70o & +66.4o
respectively. The dextrose specific rotation calculated is at 53.5o. This would mean that the prepared
dextrose solution is in line with the theoretical data. However, it should be noted that this process should
be carried out for at least several times to obtain accuracy and precision of data. In sucrose, 57.69o was
the specific rotation recorded.

Conclusion
The students were able to correlate and analyze the refractive index, angular rotation with polarimeter,
and calculated the specific rotation of dextrose, sucrose, and invert sucrose. The students were also able
to invert sucrose by performing an acid-catalyzed hydrolysis.

References:
Jim Clark (2000) Plane polarized light. Retrieved from:
https://www.chemguide.co.uk/basicorg/isomerism/polarised.html#top
Rudolph Research Analytical (2016) Refractometer uses in industry and selection. Retrieved from:
https://rudolphresearch.com/refractometers-use-selection/
John Hanson (2006) Refractoemetry. Retrieved from:
http://www2.ups.edu/faculty/hanson/labtechniques/refractometry/theory.htm