DISCUSSION

Molisch test is a chemical test you to test for the presence of carbohydrates. Molisch's reagent
is consisted of α-naphthol dissolved in ethanol. The reaction is based on the dehydration of the
sugars by sulphuric acid. The -OH group of sugars is removed in the form of water and furfural
is formed from pentose sugar while hydroxymethyl furfural is formed from hexose sugar.
These furfurals then condense with sulphonated α-naphthol to form a purple coloured product.
Polysaccharides will also give a positive reaction. In this case, the acid first hydrolyses the
polysaccharide into monosaccharides, which then get dehydrated to form furfural or its
derivatives. Results with colour other than purple indicates that there are no carbohydrates
present in the test solution. This has proven that test solutions of 1% glucose, 1% fructose, 1%
lactose, 1% sucrose and starch which give purple dye, contain carbohydrates while distilled
water which gives a yellowish solution at the end of reaction indicates no carbohydrate is
present in it. (Karki, 2018)

Benedict’s Test is a chemical test for the presence of reducing sugars. A Benedict's Solution
contains copper (II) sulphate, sodium carbonate and sodium citrate. Reducing sugars have an
aldehyde functional group which is the reducing species. When there is presence of reducing
sugars, the aldehyde functional group will reduce copper (II) ions in the Benedict’s solution to
copper (I) ions. The copper(I) oxide formed is red in colour, insoluble in water and is
precipitated out of solution. Sodium carbonate provides the alkaline conditions which are
required for the redox reaction. Sodium citrate complexes with the copper (II) ions so that they
do not reduce to copper(I) ions during storage. From the experiment, glucose, fructose and
lactose are proven to be reducing sugars as they formed brick red precipitate which is the
positive results of Benedict’s Test. Sucrose, starch and distilled water have negative results
because they do not contain the aldehyde functional group, therefore no reduction of copper
(II) ions occur. (Aryal, 2018)

Iodine test is a chemical test for the presence of starch. When iodine solution is added to starch
solution, the solution turns blue-black. This is due to the helical shape of amylose in the starch.
Iodine molecules can fit into the hydrophobic middle of the amylose helix to form starch-iodide
complex. The transfer of charge between the starch and the iodide ion changes the spacing
between the energy levels which will cause the starch-iodide complex to absorb light at a
different wavelength, resulting in a blue-black colour. When the coloured solution was heated,
it turned from blue to colourless. This is because the starch-iodide complex is unstable and
heating will disrupt the association of amylose and iodine. When the solution was cooled down
to room temperature, the blue-black solution will form once more. When sodium thiosulphate
was added, the solution turns from blue to pale blue. This is due to the reaction between
thiosulphate ion and triiodide ion to form tetrathionate ion and iodide ion.

2 S2O32– + I3– = S4O62– + 3 I–

The ions formed a colourless, therefore the solution turned from blue to pale blue. The blue
colour still present because the starch-iodide complex was in excess. (All Medical Tests, 2016)

CONCLUSION

The objective of the experiment was achieved. It is found out that carbohydrates are present in
glucose, fructose, lactose, sucrose and starch from Molisch Test. Next, glucose, fructose and
lactose are identified as reducing sugars from Benedict’s Test. Besides that, iodine test can be
used to test presence of starch.

REFERENCE

All Medical Tests. (2016). Iodine test for Starch- Its Principle, Reagents, Procedure etc.
[online] Available at: http://allmedtests.com/iodine-test-starch/ [Accessed 12 Aug. 2019].

Aryal, S. (2018). Benedict's Test- Principle, Composition, Preparation, Procedure and Result
Interpretation. [online] Microbiology Info.com. Available at:
https://microbiologyinfo.com/benedicts-test-principle-composition-preparation-procedure-
and-result-interpretation/ [Accessed 12 Aug. 2019].

Karki, G. (2018). Molisch’s Test: Objectives, Principle, Reagents, Procedure and Result -
Online Biology Notes. [online] Online Biology Notes. Available at:
https://www.onlinebiologynotes.com/molischs-test-objectives-principle-reagents-procedure-
and-result/ [Accessed 12 Aug. 2019].