School of Industrial Technology

Faculty of Applied Sciences

________________________________________________________

FST 556
Laboratory Title: Determination of Sugars
Date of experiment: 21/10/2019
Laboratory Group: Monday

Name : NUR IZZATI BINTI IDRIS

Student ID : 2019654024
Group : AS2463D

MARK:
PURPOSE

1. To determine the Refractive Index (RI) and °Brix using Refractometric method.
2. To determine the unknown sugar solution using Lane-Eynon’s Titration method.

INTRODUCTION

Carbohydrates are the most abundant and diverse class of organic compounds occurring in
nature. Chemically they are composed of carbon, hydrogen, and oxygen in the ratio CnH2nOn. Food
carbohydrates include a wide range of macromolecules that can be classified according to their chemical
structure into three major groups which is low molecular weight monosaccaride and disaccharides,
intermediate molecular weight oligosaccharides and high molecular weight polysaccharides.

Carbohydrates can be classified as simple or complex carbohydrates. Simple carbohydrates are

monosaccharides and disaccharides while complex carbohydrates are made up of many
monosaccharides such as starches and fiber (polysaccharides). According to Heroro et al., 2011,
carbohydrates are a major source of energy in the human diet with intakes ranging from 40 to 80% of
total energy requirements. Carbohydrates constitute the main source of energy for all body functions,
particularly brain functions, and are necessary for the metabolism of other nutrients. Other important
effects of carbohydrates on human physiology are satiety and gastric emptying, control of blood
glucose, insulin metabolism and serum cholesterol, and influencing colonic microflora and
gastrointestinal processes such laxation and fermentation (Herrero et al., 2011).

Sugars (glucose, sucrose, fructose, lactose and maltose), sugar polyols (sorbitol and mannitol),
oligosaccharides (GOS, galactooligosaccharides and FOS, fructooligosaccharides) and polysaccharides
(starch and non starch polysaccharides) have been described as the major classes of carbohydrates
relevant for human nutrition.

MATERIALS

Refractometer Beaker

Methylene blue Hot plate

Fehling’s solution No 1 and 2 Retort stand

Burette Filter paper

Conical flask Pipette

Volumetric flask Dropper

METHODOLOGY

Refractometric method

1. The secondary prism is opened and 2-3 drops of specimen was placed onto the centre of the
surface of the main prism. It was closed gently.
2. The measurement knob was tuned slowly while looking through the eyepiece until the boundary
line appears in the refraction field of vision.
3. The refractive index (RI) and the °Brix was read.

Lane and Eynon’s Titration method

Preparation of Fehling’s solution

1. Fehling solution was prepared by the lab assistants.

Preparation of sugar solution

1. About 10 ml of syrup solution was pipette into the 1000 ml of conical flask.
2. Distilled was filled up until the calibration mark.
3. The magnetic stirrer was put inside the conical flask to mixed the solution well.

Preliminary titration

1. About 10 ml of mixed Fehling solution was placed into the wide neck conical flask and the
flask was stand on hotplate.
2. The burette was filled with the sugar solution and the burette was clamp at the retort stand
above the flask.
3. About 15 ml of sugar solution was added to the flask that contained Fehling’s solution.
4. The flask was boiled continuously throughout the titration.
5. The solution was boiled for 15 seconds and 3 drops of methylene blue indicator
6. The sugar solution was titrated in a continuous stream of drops to the boiling mixtures until the
blue colour completely discolourised. The solution seen as a clear solution with brick-red
precipitate.
7. This procedure gave the approximate quantity of the titration.

Accurate Titration

1. The titration was repeated, all the sugar solution was added required before heating.
2. The mixture was boiled gently for 2 minutes and 3 drops of methylene blue indicator was added
and the titration was completed within a total boiling time of 3 minutes.
3. The titration was repeated to consistent volumes.
DATA AND RESULTS

Sample: Blackcurrant jam

Table 1.1: Refractive Index (RI) and °Brix of sample using Abbe Refractometer

Replicate Refractive Index (RI) °Brix Temperature

1 1.4535 65.1000 % 26.5000°C

2 1.4535 65.1000 % 26.6000°C

3 1.4536 65.1000 % 26.6000°C

Average 1.4535 65.1000 % 26.5667°C

Standard Deviation 4.714 x 10-5 0.0000 0.0471

Sample: Orange cordial

Table 1.2: Refractive Index (RI) and °Brix of sample using Abbe Refractometer

Replicate Refractive Index (RI) °Brix Temperature

1 1.3962 38.0000 % 26.7000°C

2 1.3963 38.5000 % 26.7000°C

3 1.3962 38.5000 % 26.7000°C

Average 1.3962 38.5555 % 26.7000°C

Standard Deviation 4.714 x 10-5 0.2357 0.0000

Sample: Pancake Syrup

Table 1.3: Titration of unknown sugar solution using Lane-Eynon’s Titration Method

Preliminary titration Accurate titration

Trial 1 Trial 2
Initial burette reading 0.0000 5.6000 7.7000
(ml)
Final burette reading 5.6000 7.7000 9.6000
(ml)
Volume used (ml) 20.6000 20.7000 20.5000
 (final-initial)
Average 20.6000 20.6000
Standard Deviation 0.0000 0.1

CALCULATIONS

Volume used for preliminary titration = (5.6 ml - 0.0 ml) + 15 ml = 20.6 ml

Volume used for accurate titration = (7.7 ml – 5.6 ml) + 18 ml =20.7 ml

49. 3 (Dextrose factor) x 100 = 239.32 mg of invert sugar / 100 ml

20.6 (Titration volume)

QUESTIONS

1. Define reducing sugar

A reducing sugar is any sugar that is capable of acting as a reducing agent because it has a free
aldehyde group or a free ketone group.

2. Discuss the limitations of these analyses

For physical method using abbe refractometer, these method rely on their being a change in
some physicochemical characteristic of a food as its carbohydrate concentration varies.
Commonly used methods include polarimetry, refractive index, IR, and density. This analysis
is quick and simple to carry out and can be performed with simple hand-held instruments. It is
used routinely in industry to determine sugar concentrations of syrups, honey, molasses, tomato
products and jams.

For chemical analysis using Lane and Eynon titration, these method determining the
concentration of reducing sugar in a sample. Using these analysis, the results depend on the
precise reaction times, temperatures and reagent concentrations used and so these parameters
must be carefully controlled. This titration method also cannot distinguish between different
types of reducing sugar and cannot directly determine the concentration of non-reducing sugars.
DISCUSSIONS
In this experiment, The refractive index (RI) and °Brix of the two sample which is blackcurrant
jam and orange cordial using refractometric method was determined. Refractrometer measuring the
refractive indexes of solid, liquid, and gaseous media in various regions of the optical-radiation, or light,
spectrum. If the refractive index n and its dispersion (dependence on the wavelength of the light) D are
known, it is possible to determine other quantities dependent on n and D (Morales et al., 2008). The
refractive index is correlated with the amount of soluble solids (express as sucrose concentration) using
tables, or by direct reading on the refractometer of the mass fraction of soluble solids.

Based on the table 1 and 2 the result showed the value of refractive index of blackcurrant jam is 1.4535
while for orange cordial is 1.3962. As for the °Brix reading are for blackcurrant jam is 65.1% and for
orange cordial is 38.3%. Blackcurrant jam have higher value of refractive index and higher °Brix
reading compare to cordial.

In this experiment, the concentration of unknown sugar solution using Lane and Eynon’s
titration method was determined. This method involves the determination of reducing sugars by titration
with Fehling’s solution using the redox indicator, methylene blue indicator. The classical method uses
tables indicating the amounts of invert sugar (mixture of fructose and glucose), dextrose (glucose),
fructose, maltose or lactose equivalent to volumes of reduced Fehling’s solution. This standard solution
must be specific for a particular reducing sugar. The Lane and Eynon method involves the titration of a
boiling solution of the Fehling’s solution with a dilute solution of the sugar sample.

As for experiment for titration method which is determination of total reducing sugar by using Lane-
Eynon’s Titration Method is shown in the data. The preliminary titration sample the mean for the sample
is 20.6 ml, while for the accurate titration is 20.7 ml. Based on the calculation above, we can find the
volume of dextrose per 100 ml by using the formula :

Sugar Content (mg/100mL solution) = Factor x 100

Titration (mL)

Using the above formula, the mean sugar content obtained was 239.32 mg of invert sugar / 100 ml.
CONCLUSION

The objective of the experiment was achieved which is determined the refractive index value
for blackcurrant jam is 1.4535. Meanwhile, for orange cordial the value obtained is 1.3962. Based on
result, blackcurrant jam have higher amount of carbohydrate because it have higher concentration
compare to orange cordial. High concentration of food sample show high result in carbohydrate content.
For titration method using Lane and Eynon method, the value of sugar content we get are 239.32 and
the concentration also effect the carbohydrate content.

REFERENCES
Herrero, M., Cifuentes, A., Ibáñez, E., & del Castillo, M. (2011). Advanced Analysis of
Carbohydrates in Foods, 135–164. https://doi.org/10.1201/b11218-8
Morales, V., Corzo, N., Sanz, M. L. (2008). oligosaccharide analysis to detect adulterations of honey
with sugar syrups. Food Chemistry, 107, 922–928.