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Name: Joshua B.

Dacles Date Performed: March 11,

Course & Year: BSChE 3 Date Submitted: March 19,

Experiment No. 8

I. Objectives:
1. To be able to calibrate and operate the LEICA ABBE MARK II Refractometer.
2. To be able to determine the refractive index (%solids) of sugar solution.
3. To be able to verify the relationship of the refractive index to temperature.
4. To compare experimental results with standard values of % solids of sugar

II. Theory:
The method that measures how light is refracted when it passes through a given
substance is what we called refractometry. The amount by which the light is refracted
determines the refractive index. The refractive index is a physical property that is characterize
by a pure compound. It can be used to confirm the identity of a compound and to assess its
purity by comparison with the given value.

In refractometry, the instrument that is used to obtain the refractive index of a

compound is called refractometer. Refractometers are instruments that work using the principle
of light refraction through liquids. The light passes through air in a fast manner but slows down
when it passes through liquids and this phenomenon is what gives the bent-look of the
objects when they are partially submerged in water or any liquid for that matter. In other words,
the more dissolved solids a liquid contains or shall we say water, the slower light travels
through it and therefore the more distinct the bending effect on light.
This image shows that as the pencil is submerged in the water, we see a bending look,
which in fact gives us the wrong position of the pencil in water and this is because of the
refraction of light in the water.
The general principle of refractometry states that any time a ray of light moves one
material to another at an angle, it changes direction. The ration of the angle of refraction in a
vacuum versus that in a material of interest is the refractive index of the material.

The Abbe refractometer passes light through a thin film in a sample solution, and this
illuminates the a reference mark, and movement of an adjustment knob to adjust the visual
alignment of the mark with a reference is associated with a calibration scale in order to make
the measurement on the sample solution. This refractometer measures with a precision of four
decimal places and the temperature at which the measurements is made, and remember that the
refractive index of a solution typically decreases with increasing temperature.

III. Materials/Apparatus:
A. Apparatus:
Leica Abbe II Refractometer, Analytical Balance, Beakers

B. Materials:
Distilled water, sugar, Methanol

IV. Procedure:
A. Calibration of the refractometer:
During the experiment, our group did not perform this procedure since the group that
have conducted this experiment before us had already calibrated the refractometer, therefore,
we proceeded on the next procedure.

B. Operating Instructions:
We prepared ten sugar solution samples of different concentrations expressed in %
solids. We first weighed ten different weights of sugar in the prepared beakers with the use of
the analytical balance. Then, we poured 20 ml of water in the ten beakers containing the sugars
in order to have solutions with different concentrations.
After preparing the materials, we moved on to the refractometer. We opened the prism
assembly cover and remove the protective lens tissue. Then, we cleaned the measuring prism
surface with methanol and then with distilled water and wiped it with a clean, soft tissue
because any residue left on the prism could affect the accuracy of the readings. We set the
mode selector to the desired position and applied the first sample solution on the measuring
prism surface using a dropper and closed the prism cover and positioned the illumination arm
so that the exposed face of the upper prism was fully illuminated.

Then we turned the dispersion correction wheel so that the crosshair adjustment access
hole was at the six oclock position and rotated the adjustment control counterclockwise to
position the shadow line at the bottom of the field of view.

We rotated the eyepiece to bring the crosshair into focus, and moved the shadow line to
the crosshair reticule with the coarse adjustment control. After that we rotated the dispersion
correction wheel in order to eliminate any red or green color at the edge of the shadow line and
turned the adjustment control to center the shadow line to the crosshair.

And lastly, we pushed the READ button to start the reading the values of the test
sample, such as the temperature of the sample placed on the measuring prism and other values
that will be shown on the results.

V. Diagram:
Note: see attached page on the back portion.

VI. Data and Results:

A. The data that we had gathered from the experiment are the following:

Samples Mass (g) of Volume % Solids - Refractive % Temperature % weight

sugar of water Tc index (nd) Solids (C) (Concentra
content (ml) tion)
1 1.0660 20 4.4 1.3386 3.8 28.1 5.33

2 2.0055 20 9.9 1.3467 9.3 28.2 10.03

3 3.0183 20 14 1.3532 13.4 28.3 15.09

4 4.0190 20 17.2 1.3582 16.6 28.3 20.095

5 5.0095 20 21.9 1.3660 21.3 28.4 25.05

6 6.0007 20 24.4 1.3702 23.7 28.5 30.00

7 7.0038 20 29.3 1.3788 28.7 28.7 35.02

8 8.0050 20 32.1 1.3840 31.5 28.7 40.025

9 9.0019 20 33.4 1.3863 32.6 28.7 45.01

10 10.0079 20 35.3 1.3897 34.7 28.7 50.04

B. Graph of the refractive index with concentration:

Refractive index (nd) vs Concentration










5 10 15 20 25 30 35 40 45 50

The x-axis represents the values for the concentration (wt %) of the ten sample sucrose
solutions and the refractive index for the vertical axis.

C. Graph of the refractive index with the corresponding sample solutions:










1 2 3 4 5 6 7 8 9 10

VII. Observation and Discussion of Results:

During the experiment, I observed that the refractometer is a delicate instrument and we
must have known knowledge about it before using it, since its measuring prism, the part where
drops of the sample solution is placed, is sensitive to foreign objects. Dirt and dust are the
primary enemies of the prism and thats why we cleansed the prism right after the readings of
the measurements are through for the sample.

Foreign objects must be wiped out on the prism because it may affect the readings of
the instrument and will give an inaccurate result for the sample. I noticed that the reading of the
refractometer for the % solids of the samples increases for more concentrated samples. The
same goes for the % solids-temperature compensated in which it also increases with increasing

The results shows that the reading of the refractometer for the % solids-TC is higher
compare to the results in the % solids where temperature is not considered. This means that the
temperature is a factor and has big effect on the % solids. The data also shows that the
refractive index of the samples increases as the % solids and concentration increases.
Therefore, we can say that the relationship among the values of the measurement is directly
proportional except only for the temperature. This is shown on the graph before which shows
the relationship of the refractive index on both concentration and sample solution.

In the experiment, the temperature does not vary greatly since we used the room
temperature of the sample only, we did not alter their temperature therefore, and they have
mostly likely the same temperature which is also from the reading of the refractometer.
Therefore, the data that we have obtained did not verify the theory that states: the refractive
index of a solution typically decreases with increasing temperature, since the temperature
among the sample solutions is likely the same for they are taken at room temperatures only.

VIII. Conclusion and Recommendation:

After conducting the experiment, I conclude that we were able to operate the LEICA
ABBE MARK II Refractometer and obtain the data that we needed. We were also able to
determine the refractive index of the different sample sugar solutions with the use of the

I also conclude that we were not able to verify the relationship of the refractive index to
temperature since the temperature that we used for all the sugar solutions was at room
temperature only. We have not proven whether the refractive index does really decreases as the
temperature increases. But based on the results, we can say that the refractive index of the
solutions increases with an increasing concentration, and therefore I conclude that they are
directly proportional to each other.

I recommend that the refractometer should be always handled with care and clean the
measuring prism from time to time. I also recommend for those who will be conducting this
experiment later on, that they should include in their procedure to have different temperatures
for each sample solutions, or to have different temperatures for the same solution and then test
it using the refractometer and obtain the refractive index, in order to verify if the refractive
index do decrease with increasing temperature.

IX. Applications:
Refractive index has a large number of applications. It is often applied for the
determination of identity of a particular substance, know its purity, and also measure its
concentration. It can be also used in determination of drug concentration in pharmaceutical
industry. It is applied for estimation of thermophysical properties of hydrocarbons and
petroleum mixtures.

In pharmaceutical industry, Refractometry is important in pharmacy because it can be

used as an aid in (1) identifying a pure substance, (2) establishing the purity of some
substances, (3) determining the concentrations of some simple solutions (i.e., alcohol solutions,
sugar solutions [percent of sugar in syrup]), and (4) quickly determining the uniformity of final
compounded preparations, once a refractometric standard has been established. The main
purpose of refractometry in many kinds of industry is for the quality control of their products.

X. References: