Practice No.

02

Teacher:
Victor Augusto Castro
Zavaleta

New Chimbote–Peru - 2018

 I. INTRODUCTION

Refractometry is an analytical technique that involves measuring the refractive index of

a liquid allowing the investigation of its concentration if it is a solution or its purity if
it is a unique compound and has various applications in both qualitative and quantitative aspects, in the
food analysis.
Refraction is the change in direction that a ray of light experiences when it passes from a
medium towards another of different density, due to the change in the speed of light it becomes more
It slows down the more dense the medium through which the beam passes.
The instrument used in this technique is the refractometer, which can be of different types depending on
the use and the method of measurement. The refractive index of a liquid is relative to its concentration
and for this reason, a refractometer can display the concentration in appropriate units such as
they can be ºBrix (% sugar).
A parameter that influences refractive index measurements is temperature, in which
the decrease in the refractive index with the increase in temperature is due to the decrease
of the density and dielectric constant of the medium.

Currently, the application of refractometry is important in quality control tests.

of syrups, juices, oils, as well as in the areas of research and development of new products,
from the food industry. That is why we as future engineers
agro-industrial we must have knowledge of this technique for its subsequent use since in the
Foods and beverages, the measurement of the concentration of sugar content in products is
very important as it represents a critical parameter for acceptance or rejection of our
product to be developed.

During the practice carried out, the aim was to understand the foundation of the use of the refractometer.
and its application in the determination of the refractive index (RI), as a method of analysis that
allows determining the content of soluble solids. This report presents the
collection of the obtained data and the results found after the practice carried out.

II. OBJECTIVES

Knowing the basis for the use of the instrument and the determination of the index of
refraction.
Determine the content of soluble solids in food.
Establish tabular or graphical relationships between concentration and Brix degrees, solids
solubles, total solids.
III. THEORETICAL FRAMEWORK

ABBE REFRACTOMETER: This refractometer, also based on the principle of the angle
limit. To do this, the refractometer uses the principle of refraction, which takes place in the
limit surface between the prism and the sample. The refractive index of the prism determines the limit
upper limit of the measurement range, as this must always be greater than that of the sample. The
A refractometer measures the density of liquids; the denser a liquid is, the greater it will be.
refraction.
It requires only very small amounts of the sample and gives a precision of the order of ± 2 x 10-4.
The scale is directly graduated in refractive indices for the D lines at 20° C.
It is used to measure the refractive indices (nD) of transparent or translucent liquids.
and solids being the most frequent use with transparent liquids. Calculate the percentage of sugar
content in sucrose solutions in a range of 0 to 95 degrees Brix, which corresponds
with a refractive index of 1.333 to 1.531.

Unit of Measure (Brix%)

The Measurement Scale (%) shows the percentage of concentration of soluble solids.
contents in a sample (water solution). The content of soluble solids is the total of
all the solids dissolved in water. Basically, the Brix percentage (%) is calibrated to the amount
grams of sugar contained in 100g of sugar solution.

Abbe refractometers require a water bath to control the temperature difference of the
digital refractometers in which, in these last ones, the measurement of the sample is carried out in a way
quick and simple. The digital refractive index indicator is very precise as it shows
various decimals and can be recorded electronically or sent to a printer.
 Index of refraction correction table with temperature in distilled water

Source: Abbe Refractometer Manual (2004)

Index of refraction table in distilled water at 20 °C

Food Brix degrees
water 1.3333
Source Durst, Gokel. (1985, p.124-125)

Source:
Minimum Brix degree level table for reconstituted fruit juice and puree
reconstituted
Food Brix degrees
strawberry 7.0
Peach 10
Lemon 8
Orange 11.2
Mandarin 11.2
Banana 21

Source: Codex Alimentarius Commission (2000)

IV. MATERIALS AND METHODS

4.1 Materials
Digital refractometer J157, manufactured by Rudolph Research Analytical, of
capability to identify ±0.00002 RI/±0.01 BRIX
Abbe refractometer with IR of 1 - 1.700 nD and Brix degrees of 0 - 99 %
Beakers
Mortar
Gas
Funnel
Brown sugar
Thermometer
Kitchen.
Samples: silk banana, juice orange, tangerine, strawberry jam brand
Fanny and peach juice brand Pulp.

4.2 METHODS

AOAC METHOD 932.12 for fruits and fruit products

4.2.1 Use of the instrument

The Abbe refractometer was used (See Annex No. 01) and the digital refractometer (See
Annex No. 02
For the Abbe refractometer, it was placed in a lit area, and its cover was opened.
and a few drops of distilled water were placed on the prism and the cover was closed. Then,
he observed the scale through the eyepiece and turned the measuring knob until he located the
line of demarcation of brightness and darkness in the field of vision to finally measure
the refractive index
With the digital refractometer, the cover was simply opened, and a sample was placed on the
prisma, the cover was closed and the reading was taken by pressing the digital button, giving us
the values of degrees Brix and refractive index. (See Annex No. 03)
After the measurement, the prism of each refractometer was carefully cleaned and dried.
and a drop of the substance from the problem was placed, this was to prevent any residues from remaining
that could affect future measurements.

4.2.2 Determination of the refractive index of food:

Banana and jam samples were used for the use of the Abbe refractometer, while
that for the use of the digital refractometer, samples of orange, tangerine, lemon and
peach juice. (See Annex No. 04)
In the case of the Abbe refractometer, it was placed in a lit area and a
artificial light source. It was adjusted by placing a few drops of distilled water between the prisms,
obtaining a refractive index of 1.3330.
For solid samples like the banana, it was cut into segments and placed into a mortar for
obtain the juice from the fruit and it was filtered. In the case of citrus fruits like orange, tangerine, and lemon
 they were expressed in a centrifuge tube using a gauze. And then the samples were placed in
the prism of the refractometer.
The sugar concentration of the sample (ºBrix) and the refractive index were measured and recorded.
The ambient temperature used was 20 °C

4.2.3 Effect of concentration on the refractive index:

Sucrose solutions of 5, 10, 20, 30, and 40% were prepared and the reading was taken in the
digital refractometer as described in section 3.1.1.
To determine the effect of concentration on the IR, the following was considered:
Create the n vs C graph using the least squares method and provide conclusions about it.
the dependence of n on varying the concentration C.
Obtain by interpolation the unknown concentration of a sugar solution (Cx)
knowing its refractive indexx(this is by graphical method) and also obtain Cx
analytically in the following way:

mCX b

4.2.4 Effect of Temperature on the Refractive Index.

A solution of sucrose (brown sugar) at 20% was prepared with 100 ml of water in a glass.
precipitate and was subjected to a water bath in the electric stove (See Annex No. 05) and with a
The thermometer was used to determine temperatures of 30, 40, 50, 60, and 70 ºC, and the readings were taken.
of the refractive index and Brix degrees on the refractometer.

[Link]

I.1 Use of the instrument

Table No. 01: Refractive index and Brix degrees in distilled water

Team Sample I.R. Degree Brix

Digital Refractometer Distilled water 1.33289 -0.06

Abbe refractometer Distilled water 1.333 0

The table indicates the refractive index as well as the number of degrees brix of distilled water.
determined by different refractometers, with the refractometer being the most accurate to approach the
theoretical values.

I.2 Determination of the refractive index of food:

Table N°02
 Team Sample I.R. °Brix

Distilled water 1.33289 -0.06

Lemon juice 1.3455 8.49
Refractometer
Digital Orange juice 1.35349 13.62
Tangerine juice 1.34638 9.06
Peach juice 1.34900 10.76
Distilled water 1.33300 0
Refractometer
Strawberry jam 1.46100 68
Abbot
Banana paste 1.37300 25

The table shows that there is a higher refractive index and Brix degrees in the sample of
strawberry jam and therefore a higher concentration of sugars.

I.3 Effect of concentration on the refractive index:

Table No. 02: Refractive index and Brix degrees of sucrose solutions

Concentration Of
I.R. (n) °Brix
Sucrose (C)
5% 1.34270 5.09
10% 1.34780 9.51
20% 1.35670 18.92
30% 1.36630 28.82
40% 1.37400 38.15

Graph n vs C by the least squares method

We find the slope

We find b
(∑ )(∑ )
∑ − y̅ − x̅ 1.3386
m= ( ∑ )2
0.0899
∑ 2−
 Chart No. 01: Refraction index vs
Concentration
1.38000
1.37500 y = 0.0899x + 1.3386
R² = 0.9984
1.37000
1.36500
1.36000
I.R.
1.35500
1.35000 Linear (I.R.)

1.34500
1.34000
0% 10% 20% 30% 40% 50%
c

It can be observed in the graph that the refractive index is directly proportional to the
concentration
Determination of the concentration of the substance

Formula y = 0.0899x + 1.3386

n: refractive index
Equation: Cxconcentration
n = mCx+ b
m = 0.0899
b= 1.3386

If: n = 1.36192

So replacing in the formula 25%

By interpolation

Index of 1.35670−1.36630
Concentration (C) = 0.096
refraction 0.2-0.3

20% 1.35670
X=1.36192-1.35670+ 0.02 =0.254375 = 25%
x 1.36192 0.096

30% 1.36630

I.4 Effect of Temperature on the Refractive Index.

 Table No. 04: Refractive index and Brix degrees in 20% sucrose solution subjected to
different temperatures

Temperature °C IR °Brix

30 1.3635 19.95
40 1.3625 19.42
50 1.3615 18.71
60 1.3605 18.05
70 1.3589 17.52

The table shown indicates that at higher temperatures the refractive index
decreases, the same happens with Brix degrees.

VI. DISCUSSION
Durst, Gokel (1985) and Abbe Refractometer Manual Model 315 (2004). The index of
The refraction of distilled water at 20°C is 1.333 and 1.332991 respectively.
Table No. 01 shows that the refractive index of distilled water at the temperature
at 20 °C in the Abbe refractometer gives a value of 1.333 and in the digital refractometer a
value of 1.33289.
The value of the refractive index reported by Durst and Gokel (1985) is similar to the
value of the Abbe refractometer but differs by a very minimal value compared to the value
obtained with the digital refractometer, it should be noted that the refractive index
water is sensitive to temperature variation and therefore it must be
perfectly controlled. One of the factors that affects the turning angle is the reading.
of this, since this depends on the eye of the analyst
Codex Alimentarius Commission (2000). The minimum level of Brix degrees for juice
(jugo) de fresa es 7 en durazno 10 en limón 8 en naranja 11.2 en mandarina 11.2 y en el
banana 21.
In table No. 02, the brix determination of the strawberry jam was 68, in the juice of
10.76
mandarin 9.06 and in the banana paste 25.
In the case of strawberry jam, there is a big difference compared to strawberry juice.
due to the added sugars provided to this product. Regarding the juice
peach varies very little. In the case of lemon, it is the juice that comes closest to the
value provided by the Codex Alimentarius commission (2000), while in the
orange tangerine and banana there is a greater difference, this may also be due to the
state of ripeness in which the fruit is found, as well as the reading provided by the analyst.

Rozo, González, and García (2014). The refractive index is based on determining the
solid concentration, therefore, increases in concentration will cause the index of
the concentration increases. The greater the concentration of sugars present, the greater
its density will be lower than the speed of the rays of light that pass through it, than the same
they will experience a deviation in their trajectory. This deviation in trajectory is
 directly related to the concentration of sugars and other soluble solids,
the greater its concentration, the greater the degree of deviation of the beam
of incident light and vice versa. The refractometer allows quantifying that relationship between the
degree of refraction and the concentration of sugars in different units of measurement
through the use of properly graduated scales. The Brix degrees measure the
amount of soluble solids present in a juice or pulp expressed as a percentage of
sucrose.
Graph No. 01 indicates that the refractive index is directly proportional to the
concentration, as the concentration increases, the index increases
refraction and degrees Brix.
The information provided by Rozo, González, and García corroborates the results.
obtained in practice, determining that since higher sugar concentration leads to greater
degree of sweetness and therefore a higher number of Brix degrees and refractive index.

MetAs Guide (2008). Temperature influences refractive index measurements.

for most liquids, it decreases by approximately 0.00045 when the temperature increases by 1 ºC,
in solids it decreases only 0.00001 for every 1 °C; in water it decreases
0.00010 for each 1 ºC. In general, the decrease of the refractive index with the increase
The temperature change is due to the decrease in density and dielectric constant of the medium.
Table No. 03 shows that there is an inversely proportional growth of the
temperature with the refractive index and Brix degrees, as the temperature increases
the refractive index decreases, and the same happens with the Brix degrees.
The report by Gruia MetAs (2008) reveals consistency with our results which
it indicates that correct measurements have been made regarding the analyzed foods,
but it is important to take into account

VII. CONCLUSION
The basis for the use of the refractometer and the determination of the refractive index was known.
Brix degrees. In foods such as strawberry jam, peach juice, banana, and juice of
orange, lemon, and mandarin, establishing relationships through graphs between concentration and
refractive index; it was determined that higher temperatures result in a lower refractive index and
the higher the concentration of sugars, the higher the brix degree present in the food

VIII. RECOMMENDATION

Keep in mind that the refractive index is sensitive to changes in temperature and
therefore it must be perfectly controlled for its determination.
IX. ANNEXES

Annex No. 01 Annex No. 02 Annex No. 03

Annex No. 04 Annex No. 05

X. BIBLIOGRAPHIC REFERENCE
CENAM, National Metrology Center. Querétaro, Mexico. OIML R-108. (1993).
Refractometers for the measurement of the sugar content of fruit juices. OIML,
International Organization of Legal Metrology.
Codex Alimentarius Commission (2000). Special Intergovernmental Action Group
From the Codex on Fruit and Vegetable Juices
The provided text is a URL and does not contain translatable content.
Durst. D. H., Gokel W. G. (1985). Experimental organic chemistry. Mexico: Reverte
INDEX INSTRUMENTS (1980). OFFICIAL METHODS LIST FOR
REFRACTOMETERS. England. Recovered from
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[Link]
The MetAs Guide (2008). Refraction Metrology. Jalisco, Mexico. Retrieved from
Unable to access or translate the content from the provided URL.
Manual of Refractometer Abbe 315. (2004). Recovered of
Unable to access the content of the provided URL.
[Link]
Rozo, S.; González, J.; García, D. (2014). REFRACTOMETRY AND DETERMINATION
DECONCENTRATIONS. Department of Agroindustrial Engineering Technology
Fruits Y Vegetables UNET. Venezuela. Recovered
The provided link appears to be a URL leading to a document about refractometry, but there is no specific text provided to translate.