1.

INTRODUCTION
Carbohydrates, also known as saccharides, are carbon compounds that contain
large quantities of hydroxyl groups, and are also the most important sources of
energy. They have the basic general formula Cn(H2O)nand they are the most
commonly found organic compounds in living organisms. They are classified into
several groups, namely monosaccharides, disaccharides, and polysaccharides,
depending on the number of their monosaccharide units.
Monosaccharides are further divided with regards to the number of carbons
they have pentoses and hexoses. Pentoses contain five carbon atoms while
hexoses contain six carbon atoms. They can also be classified as aldoses or
ketoses. Aldoses contain one aldehyde group while ketoses contain one ketone
group within the molecule.
An oligosaccharides monosaccharide units, on the other hand, range from two
to ten, all linked by glycosidic bonds (a covalent bond which binds between the
hemiacetal group of a saccharide). It is different from polysaccharides because it
contains multiple but few carbon atoms, whereas polysaccharides may contain up
to hundreds of monosaccharide units. Oligosaccharides and polysaccharides are
similar, however, in the fact that both of them can be hydrolysed by heating in a
slightly acidic solution.
1.1 Theories and Principles
Carbohydrates are polyhydroxy aldehydes and ketones or substances that
hydrolyze to yield polyhydroxy aldehydes and ketones. Aldehydes (CHO) and
ketones ( = CO) constitute the major groups in carbohydrates.
Carbohydrates are mainly divided into monosaccharides, disaccharides and
polysaccharides. The commonly occurring monosaccharides includes glucose,
fructose, galactose, ribose, etc. The two monosaccharides combine together to form
disaccharides which include sucrose, lactose and maltose. Starch and cellulose fall
into the category of polysaccharides, which consist of many monosaccharide
residues.
The General Tests that is used in this experiment to identify Carbohydrates are
Molischs test and Iodine Test . Molischs Test is a common test for all carbohydrates
larger than tetroses. The test is on the basis that pentoses and hexoses are
dehydrated by conc. Sulphuric acid to form furfural or hydroxymethylfurfural,
respectively. These products condense with -naphthol to form
purple
condensation product. While on the other hand, iodine test is used for the detection
of starch in the solution. The blue-black colour is due to the formation of starchiodine complex. Starch contain polymer of -amylose and amylopectin which forms
a complex with iodine to give the blue black colour.

The qualitative tests for carbohydrates which were performed for this
experiment include Fehlings Test and Bials Test. Fehlings Test forms the reduction
test of carbohydrates. Fehlings solution contains blue alkaline cupric hydroxide
solution, heated with reducing sugars gets reduced to yellow or red cuprous oxide
and is precipitated. Hence, formation of the yellow or brownish-red colored
precipitate helps in the detection of reducing sugars in the test solution. While Bials
Test is used to distinguish between pentoses and hexoses. They react with Bials
reagent and are converted to furfural. Orcinol and furfural condense in the presence
of ferric ion to form a colored product. Appearance of green colour or precipitate
indicates the presence of pentoses and formation of muddy brown precipitate shows
the presence of hexoses

1.2 Objective
To examine the reactivity of some monosaccharides, disaccharides, and
polysaccharides through series of analyses.

2. METHODOLOGY
Numerous tests have been devised for the determination of the properties
and for the determination of carbohydrates. A brief description of the most common
tests follows. Molischs test is based on the reaction of a carbohydrate with
concentrated sulfuric acid to form a furfural or hydroxyfurfural which then reacts
with the Molisch reagent ( -naphthol) to yield a colored product, generally a
purple ring at the interface of the two layers. Bials test is used to distinguish
between pentoses and hexoses. The Bial reagent contains orcinol (5-methyl
rescorcinol) dissolved in concentrated HCl with a small amount of iron (III) chloride
present. When mixed with a monosaccharide, pentoses are converted to a furfural
to yield a blue-green product. Hexoses, with prolonged heating, yield a muddy red
precipitate. Iodine reacts
with polysaccharides to give colored complexes.
Specifically amylose gives a deep blue-black color, amy-lopectin, cellulose, form red
to brown colors, and glycogen produces a reddish colored precipitate.
Monosaccharides and disaccharides are two small to trap the iodine molecules and
do not form dark colored complexes. The iodine test can be used to distinguish
between mono/disaccharides and polysaccharides. It may also be used to follow the
course of a hydrolysis reaction. In Fehlings test, reducing sugar reacts with
Fehling's reagent in alkaline medium to form an orange to red precipitate. Fehling's

reagent is commonly used for reducing sugars but is known to be not specific for
aldehydes. Positive result is detected by reduction of the deep blue solution of
cupric (II) to a red precipitate of insoluble cuprous oxide (Cu 2O). The sucrose does
not react with Fehling's reagent. Sucrose is a disaccharide of glucose and fructose.
Most disaccharides are reducing sugars.

2.1 Materials
The materials used in this experiment are the following : Test tubes, Test tube
rack, Water bath, Dropper and Beaker. While the reagents and chemicals used are
the following : Molischs reagent, Iodine solution, Fehlings reagent A&B, Bials
reagent, 95% ethanol, concentrated H2SO4, 0.01 M I2, 0.1% CHCOOH, HCl, and
concentrated HNO3.

2.2 Procedure

A. Molischs Test
In a test tube, add 2 ml of the test carbohydrate solution and 2 drops of naphthol solution. Carefully incline the tube and pour dropwise conc. H2SO4, using a
dropper,along the sides of the tube. Observe the violet colour at the junction of the
two liquids.
B. Fehlings Test
In a test tube, add 2 ml of the test carbohydrate solution and add equal
volumes of Fehling A & Fehling B and place it in a boiling water bath for few
minutes.. When the contenst of the test tube comes to boiling, mix them
together and observe any change in color or precipitate formation. The production
of yellow 'or brownish-red precipitate of cuprous oxide indicates the presence of
reducing sugars in the given sample
C. Bials Test
To 5 mL of Bials reagent add 23 mL of test solution and warm gently in a hot
water bath for 2minutes . The formation of a bluish green product is indicative of
pentoses. Hexoses generally react to form muddy brown products.
D. Iodine Test

Add 2 drops of iodine solution to about 2 mL of the carbohydrate containing

test solution.
A blue-black colour is observed which is indicative
of presence of polysaccharide.
E. Action of Alkali on Sugars
Heat 1 mL glucose solution with 1 mL 40% NaOH for 1 min. Cool and apply test
for reducing sugars (e.g.; Fehlings Test).Apply a control test with glucose solution to
observe the difference.

3. RESULTS AND DISCUSSION

Table 1. Results from Molischs and Solubility Test

SOLUBILITY TEST
SAMPLES
MOLISCHS TEST
95%
WATER
ETHANOL
1 % Solution of Glucose

Purple at the bottom

Clear

Clear

Sucrose

Purple at the bottom

Clear

Clear

Arabinose

Purple ring at the junction

Clear

Clear

Starch

Purple at the bottom

Turbid

Turbid

Glycogen

Purple at the bottom

Clear

Clear

Table 1 shows the results of Molischs test and Solubility test. Molischs test
gave negative results in glucose, sucrose, starch and Glycogen. It forms a purple
colored region at the bottom instead at a junction . The purple ring at the junction
that forms in Molischs test indicates that there is a presence of carbohydrates, it is
due to the formation of an unstable condensation product of beta-naphthol with

furfural (produced by the dehydration of the carbohydrate)

experiment, it only gives a positive results in Arabinose.

. Apparently in this

In solubility test, only starch has a result of turbid mixture both in water and
95% ethanol. As the rest of the carbohydrates (glucose, sucrose, arabinose,
glycogen) has a result of clear mixture both in water and 95% ethanol.

Table 2 Results from Qualitative Test

Samples

Fehlings test

Bials test

Action of alkali
in sugars

Glucose

no reaction

no reaction

no reaction

Sucrose

no reaction

Iodine test

Starch

(+) dark blue

Glycogen

(-) pale yellow

Forms a brown
particles in the
upper part

Agar - agar

Fehlings test is used to differentiate between reducing and non-reducing

sugars. A reducing sugar reacts with fehlings reagent in alkaline medium to form an
orange to red precipitate. A brick-red precipitate forms in the solutions containing
glucose and fructose. However in this experiment, Fehlings test shows no reaction
in glucose. Which indicates that this test was unsuccessful. While sucrose does not
react with fehlings reagent this is beause the anomeric carbon of glucose is
involved in the glucose-fructose bond and hence is not free to form the aldehyde in
solution. Based from the results, sucrose shows no reaction. This proves that
sucrose is a non-reducing sugar.

Bials test is used to distinguish between pentoses and hexoses. They react
with Bials reagent and are converted to furfural. Orcinol and furfural condense in
the presence of ferric ion to form a colored product. Appearance of green colour or
precipitate indicates the presence of pentoses and formation of muddy brown
precipitate shows the presence of hexoses. However in this experiment , glucose
shows no reaction in Bials test. This indicates the absence of Pentose in glucose.

Iodine test is used for the detection of starch and glycogen in the
solution.Iodine forms colour complexes with polysaccharides. The colour of the
complex depends of the 3d structure of the polysaccharide. Starch is a coiled
structure, which turns blue when bound to IKI (iodine solution), whereas glycogen,
which is a branched molecule, turns red/brown. In the results of Iodine test, starch
has a positive result as it gave off a color of dark blue solution. This indicates the
presence of starch. While Glycogen remains pale yellow, no characteristics colour
change is observed. This shows that there is an absence of polysaccharides or this
experiment was a failure. On the other hand, Agar-agar is insoluble for it has a
brown particles residing in upper part of the solution. Although most of the data
may not be the expected results in this study it may be due to many circumstances
of errors while performing the said experiment.

4. CONCLUSION

In General Test, only Arabinose gives a positive result. It forms a purple ring at
the junction while the others are a failure. In iodine test, starch gives a positive
result, it forms a dark blue color which indicates the presence of polysaccharides. In
solubility test, starch forms a turbid both in water and 95% ethanol.
On the other hand, Fehlings test, Bials test and Action of alkali in sugar were
unsuccessful . In this part of the experiment, all of them shows no reaction the
reason might be because of faulty procedure adopted by the observer or the
reagents for each test might have been contaminated.

5. REFERENCES
http://www.answers.com/topic/carbohydrate
http://chemistry2.csudh.edu/rpendarvis/monosacch.html

http://www.chacha.com/question/what-is-the-formula-for-glycosidic-linkageformation
http://themedicalbiochemistrypage.org/carbohydrates.php
http://answers.yahoo.com/question/index?qid=20080911213302AACHBrn