Professional Documents
Culture Documents
Experiment No. 8
Refractometer
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
solutions.
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.
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.
A. The data that we had gathered from the experiment are the following:
1.39
1.38
1.37
1.36
1.35
1.34
1.33
1.32
1.31
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.
1.39
1.38
1.37
1.36
1.35
1.34
1.33
1.32
1.31
1 2 3 4 5 6 7 8 9 10
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
concentration.
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.
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.
X. References:
http://chemistry.oregonstate.edu/courses/ch361-464/ch362/refract02.htm
https://laboratoryresource.com.au/?navaction=getitem&id=174
http://academics.wellesley.edu/Chemistry/chem211lab/Orgo_Lab_Manual/
Appendix/Instruments/Refractometry/refractometry.html
https://www.ucmo.edu/chemphys/about/documents/Refractometer.pdf
http://www.omicsonline.org/refractive-index-and-its-applications-2157-
7544.1000e117.php?aid=14768