FST556

PRACTICAL 5: DETERMINATION OF SUGARS

NAME: NUR HAZUWANA BINTI HAZUMI

MATRIC NO.: 2018200988

GROUP: AS2464A

LECTURER’S NAME: PROF. DR. NOORLAILA BT AHMAD

DATE OF EXPERIMENT: 13 MAY 2020

DATE OF SUBMISSION: 20 MAY 2020

INTRODUCTION:

Determination and quantification of sugar is important for quality control. Sugars are soluble
carbohydrate that used in food. The common types of sugar are glucose, lactose and maltose.
Sugars can occur naturally as it can be derived from plants and dairy and added to the food.
Besides, sugar can be processed using a combination of plant or dairy ingredients as a basis
(Revision of the Nutrition and Supplemental Facts Labels, 2005). The methods that will be
used in this experiment to determine the sugar are Refractometric and Lane and Eynon’s
Titration method.

Abbe refractometer will be used in Refractometric method. It is a bench-top

instrument for high precision refractive index measurement by measuring the light refracted
when it moves from the air into a sample (Berg, 2017). The brix reading indicate the actual
sugar concentration. Next, Lane and Eynon’s Titration method is to determine the
concentration of reducing sugar in sample. The reducing sugar in carbohydrate reacts with
copper sulphate and caused the indicator turn into white. A series of standard solutions of
known carbohydrate concentration is necessary as the reaction is not stoichemetric.

OBJECTIVES:

1. To determine the sugar content of strawberry jam and orange cordial by using
Refractometric method
2. To determine the invert sugar content in 100 ml of unknown sugar syrup by using Lane
and Eynon’s Titration Method

MATERIALS:

Refractometer, methylene blue, Fehling’s solution No 1 and 2, burette, conical flask,

volumetric flask, beaker, hot plate, retort stand, filter paper, pipette and dropper

METHODS:

1. Refractometric Method

The secondary prism was opened and 2-3 drops of specimen was placed onto the centre of the
surface of the main prism. It was closed gently. While looking through the eyepiece, the
measurement knob was turned slowly until the boundary line appears in the refraction field of
vision. The refractive index (RI) and the ͦBrix were read.
2. Lane and Eynon’s Titration Method
A. Preparation of Fehling’s solution

Fehling’s solution 1 was prepared by dissolving 69.28g of CuSO4.5H2O and 1 ml of 1M

H2SO4 in water and made up to 1L with water. Fehling’s solution 2 was prepared by
dissolving 346g KNaC4H4O6.4H2O and 100g NaOH in water and made up to 1L with water.
Equal volumes of Fehling’s solution 1 and 2 were mixed by pipetting a suitable volume inton
dry container. The content was mixed by swirling gently.

B. Preliminary Titration

10 ml (sugar content expected approximately 0.10-0.30g/100ml) or 25 ml (sugar content

expected approximately 0.25-0.80g/100ml) mixed Fehling’s solution was aliquot into a wide
neck conical flask and the flask was stood on hotplate. The burette was filled with sugar
solution and the burette was clamped with the burette stand above the flask. From the burette,
15 ml of sugar solution was added to the flask containing the Fehling’s solution. It was boiled
continuously throughout the titration. The solution was left to boil for 15 seconds and 3 drops
of methylene blue indicator was added. It was titrated with sugar solution in a continuous
stream of drops to the boiling mixture until the blue color was completely decolorized. It was
seen as clear solution with brick red precipitate.

C. Accurate Titration

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

1. Refractometric method

Table 5.1: Refractive index (RI) and the ͦBrix of sample using Abbe Refractometer

Sample: Strawberry jam

Replicate Refractive index (RI) ͦBrix Temperature ( ͦC)

1 1.4535 65.10% 26.50

2 1.4535 65.10% 26.50

3 1.4536 65.10% 26.50

Average±SD 1.4535±7.07x10-5 65.10±0.0 26.50±0.0

Sample: Orange cordial

Replicate Refractive index (RI) ͦBrix Temperature ( ͦC)

1 1.3962 38.00% 26.70

2 1.3963 38.50% 26.70

3 1.3962 38.50% 26.70

Average±SD 1.3962±7.07x10-5 38.33±0.2887 26.70±0.0

2. Lane and Eynon’s Titration Method

Table 5.2: Titration of unknown sugar solution using Lane-Eynon’s Titration Method

Sample: Sugar syrup A

Preliminary titration Accurate titration

Trial 1 Trial 2 Trial 1 Trial 2

Initial burette reading 0.0 0.0 6.4 7.7

(ml)

Final burette reading 6.1 5.9 8.5 9.6

(ml)

Volume of pre-added 15 15 18.5 18.5

sugar solution (ml)

Total volume of sugar 21.1 20.9 20.7 20.5

solution added (ml)

Sample: Sugar syrup B

Preliminary titration Accurate titration

Trial 1 Trial 2 Trial 1 Trial 2

Initial burette reading 0.0 0.0 12.0 14.5

(ml)

Final burette reading 7.9 7.7 14.6 17.0

(ml)

Volume of pre-added 15 15 20 20
sugar solution (ml)

Total volume of sugar 22.9 22.7 22.6 22.5

solution added (ml)
Table 5.3: Invert sugar content in 100 ml sugar syrup

Preliminary titration

Sample Invert sugar content in sugar syrup (mg/100 ml)

Trial 1 Trial 2 Average

Sugar syrup A 234.6 236.8 235.7

Sugar syrup B 525.8 530.4 528.1

Accurate titration

Sample Invert sugar content in sugar syrup (mg/100 ml)

Trial 1 Trial 2 Average

Sugar syrup A 239.1 241.5 240.3

Sugar syrup B 532.7 535.1 533.9

CALCULATION:

A. Refractometric Method

Sample: Strawberry jam

1. Refractive index
1.4535+1.4535+1.4535
Average =
3

= 1.4535

2. Brix
65.10+65.10+65.10
Average =
3

= 65.10 ͦ

3. Temperature
26.50+26.50+26.50
Average =
3

= 26.50 ͦC
Sample: Orange cordial

1. Refractive index
1.3962+1.3963+1.3962
Average =
3

= 1.3962

2. Brix
38.00+38.50+38.50
Average =
3

= 38.33 ͦ

3. Temperature
26.70+26.70+26.70
Average =
3

= 26.70 ͦC

B. Lane and Eynon’s Method

Dextrose factor
Mg invert sugar/ 100 ml = x 100%
Titration volume

A. Preliminary titration

1. Sugar syrup A

Based on 10 cm3 Fehling solution:

Dextrose factor for 21.1 ml and 20.9 ml at 21 ml = 49.5

Trial 1
49.5
Mg invert sugar/ 100 ml = x 100%
21.1

= 234.6 mg/100 ml

Trial 2
49.5
Mg invert sugar/ 100 ml = x 100%
20.9

= 236.8 mg/100 ml
 234.6+236.8
Average =
2

= 235.7 mg/100 ml

2. Sugar syrup B

Based on 25 cm3 Fehling solution:

Dextrose factor for 22.9 ml and 22.7 ml at 23 ml = 120.4

Trial 1
120.4
Mg invert sugar/ 100 ml = x 100%
22.9

= 525.8 mg/100 ml

Trial 2
120.4
Mg invert sugar/ 100 ml = x 100%
22.7

= 530.4 mg/100 ml
525.8+530.4
Average =
2

= 528.1 mg/100 ml

B. Accurate titration

1. Sugar syrup A

Based on 10 cm3 Fehling solution:

Dextrose factor for 20.7 ml and 20.5 ml at 21 ml = 49.5

Trial 1
49.5
Mg invert sugar/ 100 ml = x 100%
20.7

= 239.1 mg/100 ml
Trial 2
49.5
Mg invert sugar/ 100 ml = x 100%
20.5

= 241.5 mg/100 ml
239.1+241.5
Average =
2

= 240.3 mg/100 ml

2. Sugar syrup B

Based on 25 cm3 Fehling solution:

Dextrose factor for 22.6 ml and 22.5 ml at 23 ml = 120.4

Trial 1
120.4
Mg invert sugar/ 100 ml = x 100%
22.6

= 532.7 mg/100 ml

Trial 2
120.4
Mg invert sugar/ 100 ml = x 100%
22.5

= 535.1 mg/100 ml
532.7+535.1
Average =
2

= 533.9 mg/100 ml
DISCUSSION:

In this experiment, the purpose was to determine the sugar content of strawberry jam and
orange cordial by using refractometric method. As for the strawberry jam, the average value
of refractive index obtained was 1.4535. Meanwhile, the average value of refractive index for
orange cordial was 1.3962. Higher refractive index which means more light bends indicated
that the food sample have high sugar concentration. The high refractive index was due to the
solution that was getting thicker and created a denser medium (Mital, 2010). This indicates
strawberry jam has higher sugar concentration compared to orange cordial. Next, the brix of
the sample also can be obtained. As for the strawberry jam, the average brix value was 65.10 ͦ
and for the orange cordial was 38.13 ͦ. The degree brix was to measure the sugar dissolved in
the aqueous solution. The higher the degree of brix, the higher amount of sugar dissolved in
the food sample. Strawberry jam has higher amount of sugar compared to orange cordial.

Next, the invert sugar content in 100 ml of unknown sugar syrup need to be
determined by using Lane and Eynon’s titration method. The invert sugar calculated by
dividing the dextrose factor with the titration value and pre-added sugar. The dextrose factor
was obtained from the dextrose table. As for the sugar syrup A, in trial 1, the invert sugar
content was 239.1 mg/100 ml and 241.5 mg/100 ml in trial 2. The average value obtained
was 240.3 mg/100 ml. As for the sugar syrup B, the invert sugar content was 532.7 mg/100
ml in trial 1 and 535.1 mg/100 ml in trial 2. The average value calculated was 533.9 mg/100
ml. When all the copper sulfate in solution has reacted, any addition of reducing sugar caused
the indicator to change from blue to white. Fehling’s solution was a mixture of copper
sulphate and sodium potassium tartarate. Sodium potassium tartarate complexes with the
cupric hydroxide formed in alkaline and prevent precipitation. Reducing sugars reduced the
complex cupric hydroxide to insoluble cuprous oxide which was red color. Methylene blue
which act as oxidation-reduction indicator detected the end point of the reaction
QUESTIONS:

1. Define reducing sugar.

- Reducing sugar is a sugar that serves as a reducing agent due to its free aldehyde or
ketone functional groups in its molecular structure.
2. Discuss the limitation of these analyses.
- For the refractometric method, it is difficult to analyze materials with lower refractive
index than the prism and one must bring a possibly hazardous material close to the
eyes to read the value. For the lane and eynon’s titration method, the results depend on
the precise reaction times, temperature and reagent concentrations used. It cannot
distinguish between different types of reducing sugar and cannot directly determine the
concentration of non-reducing sugars. It also susceptible to interference from other
types of molecules that act as reducing sugar.

CONCLUSION:

In conclusion, the sugar content of strawberry jam and orange cordial was determined by
using Refractometric method. The invert sugar content that was determined by using Lane
and Eynon’s titration method in sugar syrup A was 240.3 mg/100 ml and 533.9 mg/100 ml in
sugar syrup B. Therefore, the objective of this experiment was successfully achieved.

REFERENCES:

Berg, S. (2017, April 24). Purpose of Refractometer. Retrieved from

https://sciencing.com/purpose-refractometer-5069941.html

Mital, K. S. (2010). Measuring the Effect of Sugar Concentration on the Refractive Index of.
Retrieved from http://csef.usc.edu/History/2010/Projects/J1913.pdf

Revision of the Nutrition and Supplemental Facts Labels. (2005). Washington DC: National
Academies Press.