Activity 6A

Qualitative test for

Carbohydrates

Objectives
To determine the common
carbohydrates.
To identify the significant difference
in the chemical properties of
carbohydrates.
Discuss the principle of each test.

Types of Carbohydrates
Monosaccharide is a simple sugar consist only of one unit. They serve as
building blocks for more complex carbohydrate forms.
Glucose,Fructose, Galactose
Disaccharides are a group of sugars composed of two monosaccharide groups
linked together through the loss of sugar.
Maltose, Sucrose, Lactose
Polysaccharides are complex carbohydrates composed of numerous
monosaccharides combined through the loss of water molecules.
Starch, Glycogen

Molisch Test ( Procedure )

Place 1ml of distilled H2O (control)
1 ml of 5% (glucose, galactose, maltose, fructose, sucrose, lactose,
glycogen & starch soln in each test tube
Add5 drops of Molisch Reagent
Mix content by gently shaking each tube
Incline tube while adding 1ml. conc. Sulfuric acid
Note the color of the ring formed at the junction of 2 liquids.
In the same manner add sulfuric aci to the control tube.
Record results.

Anthrone Test ( Procedure )

Mix 1ml. of glucose soln & iml. Anthrone reagent
Stand (sometime) &note color after 30 mins, dilute with glacial acetic acid or
50% H2SO4
Do the same with 5% (galactose, maltose, fructose, sucrose, lactose,
glycogen & starch)

Phenylhydrazine Test (Osazone

Formation) Procedure
In each of 8 test tube place 1ml. of 5% soln of
(#1-glucose, #2- maltose, #3-fructose, #4-galactose,
#5-lactose, #6-sucrose, #7- glycogen, #8-starch)
Add a pinch of phenylhydrazine /test tube. Mix
&stopper the test tubes (using cotton) Place them in
boiling H2O for 30 min.
Cool slowly @ room temperatue Examine crystals
formed under a microscope.

Result and Observation

Documentation

Molischs Test
A sensitive test for all carbohydrates and some compunds containing
carboydrates. Based on thedehydration of the carbohydrate by sulfuric acid
to produce an aldehyde either (furfural or a derivative) which then condenses
with the phenolic structure resulting in a purple-colored compound.

Anthrone Test
Carbohydrates are dehydrated by using conc. H2SO4 to form furfural, which
in turn condenses with anthrone to form a bluish-green complex. Sugars
react with the anthrone reagent under acidic conditions to yield a blue-green
color. Concentrated H2SO4 is a powerful dehydrating agent involved in
dehydrating sugars leading to formation of the furfural, which condenses the
anthrone to give a colored a product.

Phenylhydrazine Test (Osazone

Formation)
Osazones are formed when the sugars react with a compound known as
phenylhydrazine at boiling point.
The reaction is stepwise; first, phenyl hydrazine reacts with carbonyl group
of the sugar to form phenyl hydrazone, which then reacts with two further
molecules of phenyl hydrazine to form the osazone.

Phenylhydrazine Test (Osazone

Formation)

Analysis
In Molisch Test, from the experiment all sugar tested solutions yield positive result
since they are all carbohydrates. While the control tube which contains water give a
negative result which is a clear solution. Since its not a carbohydrate.
In the Anthrone Test, only glycogen & starch give positive results but, supposedly
all sugar solutions must give positive results in the Anthrone Test since they are all
carbohydrates but due to some factors in the process of testing them thats why
some didnt gave positive results.
In the Osazone Test, all tested sugar solutions gave a yellow-pale orange
coloration. But some sugar solutions, when they are viewed under the microscope
they didnt give expected results (crystals formation). This is maybe because other
sugar tested solutions takes longer than the time given time to form their crystals
thats why some didnt yield a positive crystals formation (glucose & sucrose), while
other yield positive results (galactose, fructose, lactose, maltose, glycogen &
starch).

Conclusion
Using Molisch, Anthrone & Osazone qualitative
test we became familiar with common principles
of carbohydrates in different qualitative tests.
We learned also their significant similarities &
differences in their reactions in different
qualitative tests. We knew also that external
factors in theprocess of testing them would
affect their reactions & results.