Name: Tinashe Katsuro

Student ID: 210304

Email: katsurot@africau.edu
Course title: Agricultural Biochemistry

Assignment

TITLE
Testing for reducing sugars and carbohydrates.

METHOD
1. Test for reducing sugars using Benedict’s Test
2. Test for carbohydrates using Molisch’s Test

OBJECTIVES
● To test the presence of reducing sugars by identifying its colour and sugar
percentage
● To to test for the presence of carbohydrates

INTRODUCTION
There are generally two types of sugar namely reducing and non-reducing sugar
based on their reducing property. Benedict’s Test is used to test for simple
carbohydrates. The Benedict’s test identifies reducing sugars (monosaccharides
and some 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 are capable of
transferring hydrogens (electrons) to other compounds, a process called
reduction. When reducing sugars are mixed with Benedict's reagent and heated, a
reduction reaction causes the Benedict's reagent to change colour. The colour
varies from green to dark red (brick) or rusty-brown, depending on the amount of
and type of sugar.Benedict’s quantitative reagent contains potassium thiocyanate
and is used to determine how much reducing sugar is present.
Molisch’s test is a chemical test which is used to check for the presence of
carbohydrates in a given analyte. This is a common test for all carbohydrates
larger than tetroses. This test is named after Czech-Austrian botanist Hans
Molisch, who is credited with its discovery. Molisch’s test involves the addition of
Molisch’s reagent (a solution of ∝- naphthol in ethanol) to the analyte and the
subsequent addition of a few drops of concentrated H2SO4 (sulphuric acid) to the
mixture.

MATERIALS
● Test tubes
● Glass syringe
● Benedict’s reagent
● Distilled water
● Dropper
● Water bath
● Test tube rack
● Concentrated Sulphuric acid
● 5 separate solutions of sugars

PROCEDURES

Benedict Test
1. 5 Test tubes were labeled as A to E
2. Each test tube was added an amount of 1 ml sugar solution
3. 2ml of benedict reagent was added to each test tube
4. All 5 test tubes were added in a water bath for 5 minutes
5. After 5 minutes, the test tubes were taken out of the water bath and the
colour changes were recorded.

Molisch Test
1. 5 test tubes were added with 1 ml of sugar solution
2. 3 little drops of iodine solution were added into each of the test tubes
 3. 2 to 3 drops of Concentrated sulphuric acid was added into the test tubes
4. The colour changes were observed and recorded
RESULTS

Test Tube Colour Sugar Percentage % Colour in Sulphuric acid

A Dark green ppt 0.1 - 0.5 White ppt

B Yellow green ppt 0.1 - 0.5 White ppt

C No colour change 0 White ppt

D Brick red ppt More than 2 Purple ring

E Orange ppt 1 - 1.5 Purple ring

DISCUSSION
Benedict test is used to determine whether the carbohydrate contains a free
aldehyde or ketone group (presence of reducing sugar). Benedict’s reagent was
added to a glucose solution and was heated in the water bath for five minutes.
After the heating, the color of test tube A changed into dark green, then there
would be 0.1 to 0.5 percent sugar in solution, this shows that the reducing sugars
are present in very low amounts. The colour of the test tube B changes color to
yellow, then 0.5 to 1 percent sugar is present, this means that the reducing sugars
are present also but in small or low amounts. The colour of solution in the test
tube D changes to Brick red, then more than 2 percent sugar is present meaning
there is a very high amount of reducing sugars presence in the solution. The
colour of the solution in test tube E changes to orange, then it means that 1 to 1.5
percent sugar is present thus having a medium amount of reducing sugars. In Test
tube C there were no colour changes meaning there were no sugars present.

In the Molisch test, a purple ring appears at the interface between the acid and
test layers in the test tube D and E which confirms the presence of carbohydrates.
The test tubes A, B and C have a negative result meaning that the solutions
remain unchanged in colour but there was a white precipitate formed inside the
test tubes. There were no carbohydrates present in them. This is because the
carbohydrates reacting with mineral acids such as sulfuric acid get dehydrated
and form furfural derivatives. These compounds react with naphthol and get
condensed to form a purple-coloured product

CONCLUSION
The presence of carbohydrates is indicated if the test solution creates purple or
purplish-red tinted layers. As a result, the Molisch test results are positive. If the
test solution does not yield purple or purplish-red tinted layers, the test solution
is devoid of carbohydrate molecules and the white precipitate is observed.

REFERENCES

https://biokamikazi.files.wordpress.com/2013/10/merged_document.pdf
Tuesday 21 February 2023, 8:30pm

https://www.abdn.ac.uk/rowett/documents/
Sourcing_Sugars_Teacher_BENEDICTS_2.pdf Thursday 23 February 2023, 5:42am

https://www.vedantu.com/chemistry/molisch-test Friday 24 February 2023,

06:23am