Abstract

Carbohydrates is the primary energy source of energy in the human body. Then Glycogen was
extracted from the chicken liver. To check if the extraction was successful, glycogen
precipitation was done by ethanol, Molishs test, iodine test and Benedicts test to indicate the
positivity of the result. The extract was then divided into portions, one for each of the following:
acid hydrolysis, enzymatic hydrolysis, Molisch's test and Iodine test. The qualitative test was
composed of the Benedict's, Barfoed's, Seliwanoff's, Bial's-Orcinol. The sample is also
hydrolysed via acidic and enzymatic hydrolysis, after which it Phenylhydrazone Test.

Introduction
Qualitative tests include Benedicts Test, Barfoeds Test, Seliwanoffs Test, Bials-Orcinol Test,
most of it produces highly coloured and visible results, and Mucic Acid Test, and Phenylhydrazone
Test are for solubility.

Benedict's test allows us to prove the presence of sugars with a free aldehyde or ketone group
(reducing sugars) . Barfoeds Test also detects the presence of reducing sugars. Seliwanoffs
Test is used to detect presence of ketoses. These are sugars containing one ketone group per
molecule. On the other hand, Bials-Orcinol Test is used to detect pentoses, sugars containing
five carbon atoms. Mucic Acid Test, named after the mucic acid, dicarboxylic, or galactaric acid
it produces after the oxidation reaction takes place, it is useful in identifying galactose.

Methodology
For Qualitative Tests for Carbohydrates

BENEDICTS, BERFOEDS, SELIWANOFFS, AND BIALS TESTS

In separate test tubes, mix 5 drops of the 0.1 M carbohydrate solutions (glucose, fructose, xylose, lactose,
sucrose, and starch) and 1 mL of the required reagent for each test. Perform one test on the different carbohydrate
solutions at the same time. Place all the test tubes at the same time into a boiling water bath. Remove the tubes from
the water bath when solutions for one test give visible colour results. Note the result and the time it took for the visible
result to form for each test.
MURIC ACID TEST
Mix 3 drops of the carbohydrate solution (galactose, lactose) and 3 drops of HNO 3 on a glass slide. Pass the
mixture over a small flame from an alcohol lamp until it is almost dry. Cool at room temperature. Examine the crystals
under the microscope and draw the mucic acid crystals. If no crystals appear, let the glass slide stand until the next
period.
PHENYLHYDRAZONE TEST
Prepare the phenylhydrazine reagent by mixing 2 g phenylhydrazine hydrochloride, 3 g CH3COONa, and 10
mL distilled water. Place reagent in a warm water bath. Stir until solution clears. In different test tubes, mix 2 drops
of carbohydrate solution (glucose, fructose, xylose, lactose, sucrose, and starch) with 4 drops of freshly prepared
phenylhydrazine reagent. Mix well and cover the tubes with cotton. Heat in a boiling water bath for 30 minutes and
record the time when yellow crystals first appear.
RESULTS AND DISCUSSIONS
Table ?. Results for Qualitative Tests for Carbohydrates

Visible Results

Carbohydrate
Solution Benedicts Test Barfoeds Test Seliwanoffs Test Bials Test
 No change in
Glucose Brick red ppt. Brick red ppt. No change in color
color
Blue-green
Fructose Brick red ppt. Brick red ppt. Cherry red ppt
solution
No change in
Xylose Brick red ppt. Brick red ppt. No change in color
color
No change in
Lactose Brick red ppt. No change/ no ppt No change in color
color
No change in
Sucrose No change in color No change/ no ppt Cherry red ppt
color
No change in
Starch No change in color No change/ no ppt No change in color
color
Glycogen
No change
Acid Hydrolysate- Blue-green solution Cherry red ppt. No change
no ppt
Enzymatic-
Unknown

Figure ? Lactose, Xylose, Fructose, Glucose, Starch, Sucrose arranged respectively

Benedicts Test
When Benedicts solution with carbohydrates are heated, the solution changes to brick red. The
reducing property of carbohydrates causes this reaction. The Copper (II) will become Copper (I)
which is indicated by the color change. The sodium carbonate provides the alkaline environment
which is required for the redox reaction. While the sodium citrate complexes with Copper (II) so
that it will not deteriorate to Copper (I) during storage.
Figure ? Fructose, Xylose, Glucose, Sucrose, Starch, and Lactose arranged respectively

Barfoeds Test
When Barfoeds reagent is mixed solution of monosaccharide or disaccharide and is heated,
crystal precipitate is formed. Copper acetate is converted to copper oxide giving a brick red
precipitate when reacted to monosaccharides and disaccharides. Disaccharides reacts slower
than the monosaccharides.

Figure? Lactose, Xylose, Starch, Glucose, Sucrose, Fructose arranged respectively

Seliwanoffs Test
When heated with this reagent, ketoses are more rapidly dehydrated than aldoses. Ketoses are
distinguished from aldoses via theirketone/aldehyde functionality. Dehydrated ketoses reacts with
resorcinol to produce a cherry red coloration, aldoses can be seen as faint pink in color.
Figure? Sucrose, Starch, Xylose, Glucose, Lactose, Fructose arranged respectively

Bials-Orcinol Test
When pentose in dehydrated by the reagent , furfural form is formed. Ocinol reacts to furfural then
reacts with iron giving the bluish colored solution.

Conclusion
Overall, the group learned about the principles and concepts about sugars, simple sugars,
reducing sugars etc. Although some results arent that accurate as it supposed to be, the group
are guided accordingly.

The group learned that Benedicts test is used for reducing sugars. Barfoeds Test also detects
the presence of reducing sugars, monosaccharides and dissacharides . Seliwanoffs Test is used
to detect presence of ketoses. These are sugars containing one ketone group per molecule. On
the other hand, Bials-Orcinol Test is used to detect pentoses, sugars containing five carbon
atoms. Mucic Acid Test, named after the mucic acid, dicarboxylic, or galactaric acid it produces
after the oxidation reaction takes place, it is useful in identifying galactose.

http://www.microbiologyinfo.com/benedicts-test-principle-composition-preparation-procedure-
and-result-interpretation/
https://allmedicalstuff.com/barfoeds-test/

https://fulltimes.wordpress.com/carbohydrates-test/

http://allmedtests.com/bials-test/