PRACTICE Nº 5

SUCROSE POLARIMETRY

I. GOALS.

 Know the fundamentals, the equipment and become familiar with the use of the
polarimeter in order to identify sucrose and other sugars

II. INTRODUCTION:

There are different instruments by which the concentration of sucrose can be

determined, in this case we will talk about the use of the polarimeter using the
polarimetry technique.

This is based on the property of the electric field vector of an electromagnetic

wave (in this case natural light) that after passing through a polarizer (glass or
sheet that allows light to pass through that vibrates in a single plane) and
subsequently a sample, if The latter has optical activity, it will produce a rotation
of the axis of the incident polarized light, which will be appreciated using an
analyzer (which is nothing more than another polarizer) on a graduated scale.

The amount of sucrose present in different commonly used substances such as

soft drinks and flavored waters has been quantified using the polarimetry
method. Polarimetry is a technique that is based on the measurement of the
optical rotation produced on a beam of polarized light when passing through an
optically active substance. An optically active substance is a chiral substance
which rotates polarized light (presents optical activity) and is measured by the
polarimeter.

III. THEORETICAL FRAMEWORK

The polarization apparatus is a demonstration instrument that is used in

conjunction with a daylight projector, for the execution of qualitative and
quantitative experiments, in front of a wide circle of listeners, in schools or
universities, to demonstrate optical activity, determination of the specific angle of
rotation and determination of concentration for known specific angles of rotation.

Functioning
The light (electromagnetic waves that are within the visible range) emerging
from the daylight projector passes through a yellow filter. According to theory,
yellow light increases measurement accuracy.

This light oscillates in different planes:

The first polar filter (polarizer) only allows the passage of one of the oscillation
planes: the light has been polarized. If a second polar filter (analyzer) is placed
behind it, rotated by 90°, the polarized light is largely absorbed since, so to speak,
the "grid" of this cross filter is in a transverse position with respect to the plane of
oscillation: maximum extinction.

If a substance is placed in the light beam (e.g. e.g.: a solution in a cuvette), and
this substance is optically active, that is, it rotates the plane of oscillation of the
polarized light to the left or right, clockwise, then the analyzer must also be
rotated to return to achieve maximum light extinction. The angle, in degrees,
between the maximum extinction without and with content in the cuvette, or
between the pure solvent and the solution, is determined by rotating the
analyzer. This angle constitutes an important measurement value, together with
the concentration of the diluted substance and the filling level of the cuvette.
From the reference point of an observer facing the analyzer, some optically
active molecules rotate the polarized light to the left (counterclockwise) and are
called levorotatory, while there are others that rotate it counterclockwise. the
right (clockwise) and are called dextrorotatory. By convention, counterclockwise
 rotation is assigned a sign (-) and a plus sign (+) is assigned to right rotation.
The amount of rotation observed in polarimetry depends on the amount of
optically active molecules in the light beam.

The measurement of the optical rotation produced on a beam of polarized light

at

passing through an optically active substance is given by the following formula.

[α]=specific rotation
α=observed rotation
c=Concentration
l=cell length (dm)

The fundamental operating parts of a polarimeter are:

 a light source (usually a sodium lamp)
 a polarizer
 a tube to keep the optically active substance (or solution) in the light beam
 an analyzer
 a scale to measure the number of degrees that the plane of polarized light
has rotated.

Some of the uses of polarimetry are in quality control, process control and
research in the pharmaceutical, chemical, essential oils, food and aroma
industries.

IV. MATERIALS AND METHODS.

3.1 INSTRUMENTAL MATERIAL AND EQUIPMENT.

3.1.1. Material to be tested, inputs and reagents, those required are:

 Commercial sucrose.
 Analytical grade glucose.

3.1.2. Equipment and instruments, the following are required.

  Balances.
 digital refractometer
 Thermometers.
 Kitchenettes.
 Erlenmeyer.
 Test tubes.
 Pipettes.
 Beakers.

V. PROCEDURE.

The procedure to follow considers the following evaluations.

Prepare aqueous solutions of 10%, 15%, 20%, 26% sucrose, and 15% glucose.

Figure 1: 10%, 15%, 20%, 26% sucrose and 15% glucose

After carefully cleaning the prism, place a few drops of the test substance. It is
recommended that the sample be transparent enough to allow light to pass
through and the temperature at 20°C (as the equipment is calibrated)

Take readings for samples at different concentrations.

 Figure 2: Measurements on the digital refractometer.

VI. RESULTS AND DISCUSSIONS.

Table 1: °Brix, rotation angle, purity and concentration of sucrose and glucose
at different concentrations.

SUCROSE SUCROSE SUCROSE SUCROSE GLUCOSE

10% 15% 20% 26% 15%

°Brix 10.2 15 20 24,7 14,7

Rotation angle 2.27 1,10 2.8 4,31 5,34

(AR)

Concentration C=2.70 C=8.27 C=28.19 C=32.39 C=50.66

(g/100 cm3) g/100 g/100 g/100 g/100 g/100
cm3 cm3 cm3 cm3 cm3
 The table presents the data in the theoretical calculations for each of the
substances.

Solution Rotation Rotation

type theoretica
l specific
10% sucrose Right- +66.5
handed
15% sucrose Right- +66.5
handed
20% sucrose Right- +66.5
handed
26% sucrose Right- +66.5
handed
15% glucose Right- +52.7
handed

DISCUSSIONS:
Sucrose is a carbohydrate, right-handed because it rotates polarized light to the
right. This is determined since by increasing the concentration the rotational
power increased.

CONCENTRATION (%) ROTATION ANGLE

10 2.7
15 8.27
20 28.19
26 32.39

Graph 1: angle of rotation vs % sucrose concentration

 Ángulo de rotacion en función de la con-
centraccion de la sacarosa(g/ml)
35
f(x) = 2.03891651865009 x − 18.3032682060391
30 R² = 0.914797427694768
25
Angulo de rotacion

20 Series2
Linear (Series2)
15
10
5
0
8 10 12 14 16 18 20 22 24 26 28
% de concentracion de soluciones de sacarosa

It can be seen that as the rotation angle increases, the concentration of optical
activity also increases.

CONCENTRATION (%) BRIX

10 10.2
15 15
20 20
26 24.7

Graph 2: angle of rotation vs % sucrose concentration

Brix vs concentraccion(%p/v)solucion sacarosa

30
25
f(x) = 0.912788632326821 x + 1.27300177619894
20 R² = 0.996965161532705
Grados brix

15 Series2
Linear (Series2)
10
5
0
8 10 12 14 16 18 20 22 24 26 28
concentracion de las soluciones de sacarosa

Brix degrees measure the total quotient of sucrose dissolved in a liquid,

 Thus, according to the graph, the value of the degrees Brix is directly
proportional to the concentration of sucrose present in the samples, that is, as
the concentration of sucrose increases, the value of the degrees Brix increases,
which is correct.
This graph is governed by the equation:

y = 0.9128x + 1.273
R² = 0.997

VII. CONCLUSIONS

 The fundamentals and the equipment to identify sucrose and other sugars were
known.
 In this case, a polarimeter was used to measure the rotation angle of sucrose
and glucose at different concentrations.
 The temperature at which the substance is found, the wavelength, the
concentration, the elapsed time, the presence or absence of catalysts, are some
factors that delay or accelerate the inversion of sucrose into its simplest
carbohydrates. Measurements of Optical rotation can be used to determine the
concentration and/or purity of a substance, or simply to detect the presence of
an optically active chemical substance in a mixture. Errors may occur in the
measurement of the human-type rotation angle when the presence of bubbles
remains in the observation or instrumentation tube due to it being poorly
calibrated.

VIII. RECOMMENDATIONS

 The samples used must be kept in adequate conditions.

 Avoid clutter in the counter where the practice is carried out and work cleanly.

IX. BIBLIOGRAPHY

 DOUGLAS A. SKOOG,F. JAMES HOLLER, STANLEY R. CROUCH. (2008).

PRINCIPLES OF INSTRUMENTAL ANALYSIS. MEXICO DF: CENGAGE
LEARNING.
 DOUGLAS A. SKOOG,F. JAMES HOLLER, STANLEY R. CROUCH. (2005).
FUNDAMENTALS OF ANALYTICAL CHEMISTRY. MEXICO DF: CENGAGE
LEARNING.
 W.F. PICKERING. MODERN ANALYTICAL CHEMISTRY. REVERTÉ
EDITORIAL
 Allier Rosalía etal THE MAGIC OF CHEMISTRY EPSA editorial, Mexico City
1995 1st edition.
 https://www.3bscientific.es/product-manual/U14390_ES.pdf .

ANNEXES

Calculations

[ α ]20D = α∗100
c∗L

α∗100
c=
[ α ]20D ∗L

Where:

[ α ]20D = Specific angle of rotation with D line of Na and 20°C.

α = Measured angle of rotation (scale reading)

c = Concentration of the solution in (g/100cm 3 )

L = Layer thickness (filling height) in dm.

 Finding the Concentration for 10% Sucrose

[ α ]20D = 66.52° (Values taken from Rapoport/Raderecht, Physiologisch-

chemisches Praktikum, VEB Verlag Volku. Gesundheit, 1972)
α = 0,36°
c=?
L = 0.2 dm
20 α∗100
[ α ]D =
c∗L
 0 , 36 °∗100
∗dm∗g
0.2
C=
66.52 ° cm3

C=2.70 g/100 cm 3

 Finding the Concentration for 15% Sucrose

[ α ]20D = 66.5° (Values taken from Rapoport/Raderecht, Physiologisch-

chemisches Praktikum, VEB Verlag Volku. Gesundheit, 1972)
α = 1.10°
c=?
L = 0.2 dm.
[ α ]20D = α∗100
c∗L

1.10 °∗100
∗dm∗g
0.2
C=
66.52 ° cm3

C=8.27 g/100 cm 3

 Finding the Concentration for 20% Sucrose

[ α ]20D = 66.5° (Values taken from Rapoport/Raderecht, Physiologisch-

chemisches Praktikum, VEB Verlag Volku. Gesundheit, 1972)
α = 3.75°
c=?
L = 0.2 dm.
[ α ]20D = α∗100
c∗L

3.75 °∗100
∗dm∗g
0.2
C=
66.5 ° cm3

C=28.19 g/100 cm 3

 Finding the Concentration for 26% Sucrose

 [ α ]20D = 66.5° (Values taken from Rapoport/Raderecht, Physiologisch-
chemisches Praktikum, VEB Verlag Volku. Gesundheit, 1972)
α = 4.31°
c=?
L = 0.2 dm.
[ α ]20D = α∗100
c∗L

4.31°∗100
∗dm∗g
0.2
C=
66.52° cm3

C=32.39 g/100 cm 3

 Finding the Concentration for 15% Glucose

[ α ]20D = 52.7° (Values taken from Rapoport/Raderecht, Physiologisch-

chemisches Praktikum, VEB Verlag Volku. Gesundheit, 1972).

α = 5,34°
c=?
L = 0.2 dm.
[ α ]20D = α∗100
c∗L

5 , 34 °∗100
∗dm∗g
0.2
C=
52.7 ° cm3

C=50.66 g/100 cm 3