EXPERIMENT NO : 3

TITLE OF EXPERIMENT : DETERMINATION OF REDUCING SUGAR

NAME : NURIQMAL HAFIDZ BIN ZAMBRI

ID NUMBER : 2019434184

LECTURER’S NAME : MADAM DR. SHARIFAH RAINA

EXPERIMENT DATE : 26 SEPTEMBER 2019

SUBMISSION DATE : 2 OCTOBER 2019

 Introduction
To determines the presence of reducing or non-reducing sugar in a test solution
we can use benedict’s test. The benedict’s solution is the principal reagent in benedict’s
test for reducing sugars that contains copper (II) sulphate, sodium carbonate and sodium
citrate.
There are two classes of sugars, reducing or non-reducing based on their ability to
act as reducing agent during the benedict’s test. During a redox reaction and itself oxidized,
a reducing agent donates electrons. The reducing agent in reducing sugars is the aldehyde
functonalk group. Reducing sugars have either an aldehyde functional group or have a
ketone group (which can convert into an aldehyde in an open chain form).
Reducing sugars are simple sugars and include all monosaccharides and most
disaccharide, some examples of monosaccharide are glucose,fructose and galactose.
Example of reducing disaccharide are lactose and maltose. The disaccharide sucrose is not
a reducing sugar. In fact of sucrose is the most common non-reducing sugar.
So the question is, is it the benedict’s test for reducing sugars qualitative or
quantitative? The test may be qualitative, or it may be quantitative.
The qualitative test produces a colour change from blue to green to yellow to
orange to brick red. The qualitative test is also regarded as semi quantitative as the colour
obtained correlates to the concentration of reducing sugars in the solution. This allow for a
rough estimation of the amount of reducing sugar present.The quantitative test involves the
use potassium thicyanatewband the production of copper thiocyanate as white or pale
green precipate. This precipate can then be titrated.
Benedict’s test is used to test for simple carbohydrates. The benedict’s test
identifies reducing sugars (monosaccharide’s and xsome disaccharides), which have free
ketone or aldehyde functional groups. Benedict’s solution can be used to test for the
presence of glucose in urine.
Some sugars such as glucose are called reducing sugars because they sare
capable of transferring hydrogens (electrons) to other compounds, a process called
reduction. When reducing sugars are mixed with Benedicts reagent and heated, a
reduction reaction causes the benedicts reagent to change color. The color varies from
green to brick red, depending on the amount of and type of sugar.
Materials

1. Test tube
2. Test tube rack
3. Test tube holder
4. Measuring cylinder 10 ml
5. Benedict’s solution
6. Distilled water
7. Glucose solution
8. Sucrose solution
9. Starch solution
10. Water bath
11. Masking paper tape
12. Marker pen

Procedure

1. Instructor give a briefing on the lab to the student.

2. Place four test tube in a test tube rack and label with tube 1, tube 2, tube 3 and tube
4.
3. Each tube must filled with 2 ml of each sollution
a. Distilled water ( tube 1)
b. Glucose solution ( tube 2 )
c. Sucrose solution ( tube 3 )
d. Starch solution ( tube 4 )
4. Add 2 ml of Benedict’s solution to each test tube. Swirl the test tube to mix the
solution. Record the initial colour of all solution.
5. Place all the test tube in a boiling water ( water bath ) for three to five minutes or
until there is no more colour changes.
6. Remove the test tube from the water bath using test tube holder and place them in
the test tube rack. Record the colour of each tube.
7. Once the experiment is done, discard the content of the test tube and rinse them
properly and dry the test tube.
Results: Below are the table and picture of benedict’s test for reducing sugars.

Observation Interpretation

No Colour Change ( Blue ) No reducing sugars present

Green Trace amounts of reducing sugars

present
Light Green Low amounts of reducing sugars
present
Brick Red Large amounts of non-reducing sugars
present

Table 1: The observation and interpretation of benedict’s test for reducing sugars

Figure 1: Distilled water that was mixed with 2 ml benedict’s solution (tube 1)
Figure 2: Glucose solution that was mixed with 2 ml benedict’s solution ( tube 2 )

Figure 3: Sucrose solution that was mixed with 2 ml benedict’s solution ( tube 3 )
Figure 4: Starch solution that was mixed with 2 ml benedict’s solution ( tube 4 )

Figure 5: All the test tube are heated in a beaker of boiling water
 Figure 6: There are color changes in some test tubes

Figure 7: Shows the results after the test tube is soaked in boiling water for 4 minutes
Discussion

So after we finished with this experiment which is to determination of reducing sugar

we found that when benedict’s solution and simple carbohydrates are heated, the solution
changes to brick red color. This reaction is caused by the reducing property of simple
carbohydrates. The copper (II) ions in the benedict’s solution are reduced to copper (I) ions,
which causes the color change.
The red copper (I) oxide formed is insoluble in water and is precipated out of
sollution. This accounts for the precipate formed. As the concentration of reducing sugar
increases, the nearer the final color is to brick red and the greater the precipate formed.
Sometimes a brick red solid color, copper oxide, precipates out of the sollution and collects
at the bottom of the test tube.
Sodium carbonate provides the alkaline conditions which are required the alkaline
conditions which are required for the redox reaction. Sodium citrate complexes with the
copper (II) ions so that they do not deteriorate to copper (I) ions during storage.
Complex carbohydrates such as starches DO NOT react positive with the benedict’s
solution test unless they are broken down through heating or digestion ( try chewing
crackers and then doing the test ). Table sugar ( disaccharide ) is a non-reducing sugar and
does also not react with the iodine or with the benedict’s reagent. Sugar needs to be
decomposed into its components glucose and fructose then the glucose test would be
positive but the starch test would still be negative.
Benedict’s solution is a deep-blue alkaline solution used to test for the presence of
the aldehyde functional group,-CHO. One litre of benedict’s solution can be prepared from
100g of anhydrous sodium carbonate, 173g of sodium citrate and 17.3g of copper (II)
sulfate pentahydrate.
 Conclusion

Reducing Sugars have an aldehyde functional group which can reduce soluble
copper (II) ions - in copper (II) sulphate - to insoluble copper (I) ions - in copper (i)oxide.
The copper (I) oxide is seen as a precipitate. If the color upon boiling is charged into green,
then there would be 0.1 to 0.5 percent sugar in solution. If it changes color to light green,
then 0.5 to 1 percent sugar is percent and if it changes to brick red color, it means that
more than 2 percent sugar is present in solution.

 Positive Benedict’s Test: formation of a reddish precipate within three

minutes. Reducing sugar present. Example: Glucose
 Negative Benedict’s Test: No color change ( Remains Blue ). Reducing
sugars absent. Example: Sucrose.
Task

 Indicate the initial and final colur of each test tube

i. Test tube 1
From blue to blue (no colour change)
ii. Test tube 2
From blue to light green
iii. Test tube 3
From blue to green
iv. Test tube 4
From blue to brick red colour

 Determine the reducing sugar and non-reducing sugar.

Observation Interpretation

Test Tube 1 ( Distilled Water) No reducing sugars present

Test Tube 2 ( Glucose Solution) Trace amounts of reducing sugars

present
Test Tube 3 ( Sucrose Solution) Low amounts of reducing sugars
present
Test Tube 4 ( Starch Solution) Large amounts of non-reducing sugars
present
 Explain how benedict’s solution able to determine the reducing sugar

When benedict’s solution and reducing sugars are heated together, the
solution changes its colour to brick red colour. This colour is due to the presence of
simple carbohydrates. In specific, the copper (II) ions in benedict’s solution are
reduced to copper (I) ions, which causes the colour change.
Sugar are classified as reducing or non-reducing based on their ability to act
as a reducing agent during the benedict’s test . reducing sugars have either an
aldehyde functional group or have ketone group-in an open chain form- which can
be converted into an aldehyde.
References
Fleischer, H. (2019). The Iodine Test For Reducing Sugars. World Journal Of Chemical Education, 50-
57.

Robert , D. S., Robert, L. H., & Vaughan, M. (2016). Benedict's Solution, Reagent For Measuring
Reducing Sugar. The Clinical Chemistry Of Stanley R. Benedict, 10-17.

Zerban, F., Naquin, W. P., & Chem, A. J. (2007). Contribuition From The Laboratary Of The Louisiana
Sugar Experiment Station. On The Determinanation Of Reducing Sugar, 49-52.
Lecturer give a briefing to Record the initial colour of
her student in lab. all solution.

Discard the content of the

Place four test tube in a test test tube and rinse them
tube rack. properly

Label with tube 1, tube 2, Dry the test tube by

tube 3 and tube 4 using a flattening the test tube in
masking paper tape. revers on the test tube rack.

Add 2 ml of Benedict’s
solution to each test tube.