INTRODUCTION

A refractometer is an instrument used to measure the refractive index of gases,

liquids,

and

translucent

solids

like

gemstones.

The

refractive

index

is

measurement for how much the speed of light is reduced in a solution, gas, or solid.
A shadow line appears inside the refractometer and is created by prisms and lenses.
The user will view the shadow line through a magnifying eye piece. Sugar analysis
includes the analysis of sucrose and other sugars in sugar processing and
commercial trade, and in sugar-containing foodstuffs. In sugar production and sugar
trading, sugar analysis also includes the determination of other components present
in the sugar.
Physical method for sugar analysis is through refractometric method. The
refractometric value of sugar solutions is used as a rapid method for the
approximate determination of the solids content (also known as dry substance),
because it is assumed that the non-sugars present have a similar inuence on the
refractive index as sucrose. Measurement is usually carried out on a Brix
refractometer, which is graduated in percentage of sucrose on a wt/wt basis (g
sucrose/100 g solution).Tables are available that give mass fraction corrections to
refractometric values at temperatures different from 20C. All chemical compounds
have an index of refraction. Refractive index can be determined by measuring the
angle of incidence to angle of refraction.
Analysis method for determination of sugar is lane-eynons titration method. It is
titration method based on reaction of reducing sugar with copper sulphate followed
by reaction alkaline titrate. Mixture boiled for specific time, followed by addition of
methylene blue. The constant volume modification, a more recent change to the
method, has allowed for greater standardization, increased sensitivity, and the use
of a simple formula instead of tables to determine the amount of invert. The method
determines reducing sugars in the presence of sucrose, and is used for raw cane
sugar, cane processing products, and specialty sugars having low levels of invert.
The higher the value of brix and RI the higher the sucrose content in food.

I.

REFRACTOMETRIC METHOD

OBJECTIVE
To determine the total soluble solids of selected food samples using refractometric
method.

PROCEDURE
1. Secondary prism is opened and the specimen is dropped onto the surface of
the main prism. The secondary prism is then closed gently.
2. The measurement knob is turned slowly until the boundary line appears in
the refraction field of vision.
3. The refractive index and Brix is read.

RESULT

SAMPLE

TRIAL

BRIX
SUCROSE

APPLE
JUICE
HONEY

GLUCOSE

1
2
3
MEAN
1
2
3
MEAN
1
2
3
MEAN

12
11.2
11.2
11.47
78
77.8
78.2
78
0.2
0.2
0.2
0.2

INDEX TC
1.3503
1.3503
1.3503
1.3503
1.4948
1.4948
1.4943
1.4946
1.334
1.334
1.3337
1.3339

REFRACTIVE INDEX
SUCROSE SUCROSE
SOLID
10.9
10.9
10.9
10.9
81
81
80.8
80.9
0
0
-0.2
-0.067

SOLID TC
11.6
11.6
11.6
11.6
81.6
81.6
81.4
81.53
0.7
0.7
0.5
0.63

SUCROSE
INDEX
1.3492
1.3492
1.3492
1.3492
1.4934
1.4934
1.4928
1.4932
1.333
1.333
1.3327
1.3329

DISCUSSION
From the experiment, refractometer is used to determine the total soluble solids of
selected food, which are apple juice, honey and glucose. The index of refraction is

the degree of deection of a beam light that occurs when it passes from one
transparent medium to another.

The refractive index of food samples can be

determined by measuring the angle of incidence to angle of refraction. Most

handheld refractometers give the concentration of the dissolved substance in Brix
(Bx), a scale defined in terms of sucrose content. Meanwhile, the value of refractive
index can be determined by using Abbe refractometer.
Through the experiment, the value of brix and refractive index is determined where
apple juice, honey and glucose brix value is 11.47, 78 and 0.2 respectively. The brix
value is determined by measured the sugar content in the food samples as it has
been used to measure the measurement of sugar in grape and other fresh fruit,
processed food, sugar solutions and beverages. From the brix value, a food sample
which is honey show highest brix value among all the food samples which indicates
higher content of sugar in honey. Glucose then shows the lowest brix value than
honey and apples juices.
Meanwhile, the refractive index value is measured by sucrose index TC, sucrose
solid, sucrose solid TC and sucrose index. The mean value for refractive index in
apple juice in sucrose index TC is 1.3503, sucrose solid 10.9, sucrose solid TC 11.6
and sucrose index is 1.3492. While the refractive index value in honey for sucrose
index TC is 1.4946, sucrose solid 80.9, sucrose solid TC 81.53 and sucrose index
1.4392. Besides, sucrose index TC, sucrose solid, sucrose solid TC, and sucrose
index in glucose show the value which is 1.3339, -0.06, 0.63 and 1.3329
respectively. The result show honey has highest refractive value than apple juices
and glucose. In addition, glucose shows lower refractive value among the food
samples.
In this experiment, there are some sources of error that we did that may affect the
accuracy of the results during conducting the experiment. For example, the eyes is
not parallel to the scale while read the brix value in handheld refractometer and the
abbe refractometer is not calibrated properly as the past sample may interfere the
reading of new samples. To minimize these errors various precautions should be
taken such as, three consecutive of weighing samples were carried out to eliminate
the errors of the handlers.

CONCLUSION
The brix value for Apple juice, honey and glucose is 11.47, 78 and 0.2 respectively.
The refractive index value for honey show highest refractive value than other food
samples which is 1.4946 for sucrose index TC, 80.9 for sucrose solid, 81.53 for
sucrose solid TC and 1.4932 for sucrose index. Thus, the hypothesis is accepted.

II.

LANE EYNONS TITRATION METHOD

OBJECTIVE
To determine the concentration of unknown sugar solution using Lane and Eynons
titration method.
PROCEDURE
PREPARATION OF CHEMICAL REAGENTS
-

Fehlings solution A and B is mixed equally by pipetting process.

Methylene blue is prepared

PRELIMINARY TITRATION
1. The burette is filled with sugar solution.
2. 10 mL of mixed Fehlings solution is pipetted into a conical ask and 10 15
3.
4.
5.
6.

mL of sugar solution from the burette is added.

The ask is heated on the hotplate and the liquid is boiled briskly.
4 drops of 1 % methylene blue is added and the titration is continued.
The titration was further while the liquid is boiled.
The sugar solution is continuing added while being boiled until the color is
discharged. ( A )

ACCURATE TITRATION
1. 10 mL of mixed Fehlings solution is pipetted into a conical ask. ( A- 2 mL )
of sugar solution from the burette is added.
2. The ask is swirled round, while the ask is being heated and boiled for 1 2 min. 4 drops of methylene blue solution is added.
3. About 2 3 drops per second of sugar solution is added and the titration is
completed within 3 min from the time boiling commences.
4. The dextrose table is used to calculate the concentration of sugar in the
solution.

CALCULATION

Sugar Content (mg / 100 mL solution) =

Factor x 100

Titration (mL)
=

51.1 x 100
50 mL

102.2 mg / 100 mL solution

RESULT
SAMPLE
PRELIMINARY
TITRATION
ACCURATE
TITRATION

TRIAL
1

mL
50 Ml

1
2
3
MEAN

50
50
50
50

Ml
Ml
Ml
Ml

FACTOR
51.1
51.1
51.1
51.1
51.1

DISCUSSION
From the experiment, we need to determine the concentration of unknown sugar
solution by using Lane and Eynons titration method. Lane Eynons titration method
is an example of a titration method of determining the concentration of reducing
sugars in a sample. A burette is used to add the carbohydrate solution being

analyzed to a ask containing a known amount of boiling copper sulfate solution

and a methylene blue indicator. The reducing sugars in the carbohydrate solution
react with the copper sulfate present in the ask. Once all the copper sulfate in
solution has reacted, any further addition of reducing sugars causes the indicator to
change.
Based on the result recorded, the volume of sugar solution recorded for the
indicator to decolorize is 50 mL. The factor also can be determined by referring to
dextrose table which is 51.1 for 10 cm3 of Fehlings solution.

The solution is

changes from blue to brick red which indicate the reaction of reducing sugar with
copper sulphate followed by reaction of alkaline tartrate. In addition, the total
reducing sugar can be determined by calculate the sugar content by using this
formula:Sugar Content (mg / 100 mL solution) =
Factor x 100
Titration (mL)
=
=

51.1 x 100
50 mL
102.2 mg / 100 mL solution

Through the experiment , there are some source of error that we did that may affect
the accuracy of the result such as the mixed Fehlings solution is not thoroughly
heated while the titration process with sugar solution is conducted. Besides, the
indicator is also added before the mixed Fehlings solution is heated. This may
interfere the result. To minimize these errors various precautions should be taken,
care during conducting this experiment must be employed besides three
consecutive of titration and preliminary titration were carried out to eliminate the
errors of the handlers.

CONCLUSION

The concentration of unknown sugar solution can be determined by using Lane and
Eynons titration method which is 102.2 mg / 100 mL solution. Hence , the
hypothesis is accepted.

QUESTION
1. What are reducing sugar?
Reducing sugars are containing an aldehyde group. These sugars can be
oxidized to form carboxylic acid and have an open-chain compound structure.
Reducing sugars contain either a free aldehyde group or a free ketone group.
Both ketone and aldehyde are organic compounds that contain a carbonyl
group within the chain. A reducing sugar that contains an aldehyde group is
called an aldose; one with a ketone chain is a ketose.

2. What is refractive index?

The refractive index or index of refraction of a substance is a measure of the
speed of light in that substance. It is expressed as a ratio of the speed of light
in vacuum relative to that in the considered medium. The velocity at which
light travels in vacuum is a physical constant, and the fastest speed at which
energy or information can be transferred. However, light travels slower
through any given material, or medium, that is not a vacuum.
3. Write the principle reaction (chemical reaction) that occurs in the Lane
Eynons determination of reducing sugar.
The principle that occur in Lane- Eynons determination of reducing sugar is
based on the reaction of reducing sugar with a solution of copper sulfate
followed by reaction with alkaline tartrate. Mixture is boiled for specific time,
followed by addition of methylene blue as indicator. Lastly, the colored
solution is titrated until discoloration of the indicator.
4. Why is the sample boiled during the titration?
The sample is boiled during the titration to prevent oxidation and to convert
alkaline cupric tartrate to insoluble cuprous oxide.

REFERENCES
K. Othmer , (2010), Sugar Analysis , Encyclopedia of Chemical Technology , John
Wiley & Sons, Inc.
P.

Mellet , ( 1980 ) , SOME

TITRATION

METHOD

PRODUCTS

FACTORS

AFFECTING THE

LANE

FOR DETERMINING REDUCING SUGARS

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from

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http://www.sasta.co.za/wp-

content/uploads/Proceedings/1980s/1980_Dunsmore_Some%20Factors%20Affecting
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(2003) , What is a Refractometer? , Retrieved from http://www.wisegeek.com/whatis-a-refractometer.htm
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