Alquiza, Angel Shen P.

Balmori, Christian Rey H.

Chem 153 YC Lab
Experiment 5: Analysis of Carbohydrates

Carbohydrates (also called saccharides) are molecular compounds made from just
three elements: carbon, hydrogen and oxygen. Monosaccharides (e.g. glucose) and
disaccharides (e.g. sucrose) are relatively small molecules. They are often
called sugars. Other carbohydrate molecules are very large (polysaccharides such as
starch and cellulose).

Carbohydrates are a source of energy for the body e.g. glucose and a store of
energy, e.g. starch in plants. It is a building blocks for polysaccharides (giant
carbohydrates), e.g. cellulose in plants and glycogen in the human body. And
components of other molecules like DNA, RNA, glycolipids, glycoproteins, ATP (Royal
Society of Chemistry, 2004).

A. General Test for Carbohydrates

This experiment aims to introduce you with the identification of unknown

carbohydrates. To gain maximum benefit, observations should be related, as far as
possible, to the structure of the substances examined.

Molisch Test

In the general test for Carbohydrates using the sample tests; Glucose, Sucrose,
Arabinose, Starch and Cotton suspension, in testing the Molisch and Anthrone
reagents, it was observed by the color and through H2SO4. In Molisch test, it gives a
positive reaction since the sample used were carbohydrates creating a purple ring in
the middle of solution.

All sample has a purple ring in the middle and a black suspension beneath, and
the differences were the solution located above the purple ring. It was observed and
can be seen the picture above that Glucose created a pale white-pink, sucrose for red-
pink like color and its purple is somehow mixed with the black suspension, Arabinose
for pale pink, Starch for pink and Cotton suspension for pale red-pink color.

Anthrone Test

For the Anthrone test, all sample resulted positive except the cotton
suspension. It was positive because the reaction the reagent with the H2SO4 resulted
to green to dark blue-green results. The cotton suspension was exempted because it
does not create a green nor blue-green sample, instead it was yellow as shown above.

B. Iodine Test for Polysaccharides

Iodine (Iodine-Potassium Iodide, I2 KI) staining distinguishes a polysaccharides

such as starch ((C6H10O5)n) from monosaccharides, disaccharides, and other
polysaccharides. The sole purpose of this test is to determine the presence of starch.
The basis of this test is that, starch is a coiled polymer of glucose. Iodine fuses with
these coiled molecules and that results the appearance of bluish black color. There
are non-coiled molecules or non-coiled carbohydrates that doesn’t interact or fuse with
Iodine. In this experiment, we had four (4) solutions: starch, dextrin, gum arabic, and
agar-agar. We were able to mix Iodine with these solutions, unfortunately, starch is
the only solution that appeared to be bluish black and the rest were turned into red
orange (dextrin) and yellow (gum arabic and agar-agar). Therefore, there are no
presence of starch for those solutions who didn’t turned into bluish black color.
Iodine solution fused with four solutions in a spot plate

C. Qualitative Test for Sugars

This test is also known as Analysis of an Unknown Sugars.

For Benedict’s, Barfoed’s, Seliwanoff’s, and Orcinol’s Test

I. Benedict’s Test

This test is used for simple carbohydrates. Benedict’s test determines reducing sugars
(monosaccharides and some disaccharides), that contains an aldehyde or ketone
group. 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 Benedicts reagent to change color (Aryal, 2019). In this
test, we had seven solutions: Glucose, Fructose, Arabinose, Maltose, Sucrose,
Galactose, and Lactose. In this experiment, our Glucose formed a reddish precipitate
within 3 minutes. This means that, reducing sugars are present in a glucose. Our
Sucrose here remained blue, that means, reducing sugars are absent in a Sucrose.

II. Barfoed’s
Test
 This test is used to detect the reducing sugars and to distinguish
monosaccharides from reducing disaccharides. According to Gaurab Karki (2018),
Barfoed’s test is used for distinguishing monosaccharides from reducing
disaccharides. Monosaccharides usually react in about 1-2 minute while the reducing
disaccharides take much longer time between 7-12 minutes to react with the reagent.
Brick red color is obtained in this test which is due to formation of cuprous oxide. A
presence of brick red color precipitate signifies a positive Barfoed’s test. Whereas, an
absence of brick red color would say that it is a negative Barfoed’s test. In our
experiment, our Lactose and Sucrose had no brick red color precipitate, and therefore,
they are disaccharides, and the rest were monosaccharides since they have produced
brick red precipitate (Glucose, Fructose, Arabinose, Maltose, and Galactose).

III. Seliwanoff’s Test

This test is a compound test that separates aldose and ketose sugars.
According to Nicolenk (2013), it is specific to identifying ketoses since the reaction
occurs faster with ketoses. Aldose sugars also react but at a much slower rate. Hence
distinction between a ketose and aldose sugar may be performed. In this experiment,
our Glucose and Maltose appears to be cherry-red precipitate that is formed in the
reaction indicating the presence of the ketose sugar. The rest indicates the presence
of an aldose sugar.
IV. Orcinol’s Test

This test is used to detect the

presence of furanoses (five-
membered rings). These sugars, or
compounds containing them, react with Bial's reagent to give a green or olive colored
solution. Furan rings contain five carbons but sugars with furan rings can contain more
carbons outside the ring, and all sugars with a furan ring will react in Bial's test. Yellow
color turns greenish indicates if sugar is a furanose (has a five membered ring like
furan). If sugar is a pentose-furanose: color turns green/olive; a hexose (or higher)-
furanose: color turns a muddy brown, or if the sugar is a pyranose: no change in color...
In our experiment, it appears that our Glucose, Maltose, Galactose, Lactose, and
Arabinose had no change of color and we conclude that these are pyranose. Our
Fructose and Sucrose appears to be an olive color precipitate and thus, it is a pentose-
furanose sugar.

V. Mucic Acid Test

Mucic Acid Test is a test used to identify the presence of the sugar galactose
in food or in synthetics manufacture. When concentrated nitric acid is heated together
with galactose, a dicarboxylic acid called mucic acid forms as a white precipitate,
which counts as a positive result. Its principle/s is/are, Oxidation of most
monosaccharides by nitric acid yields soluble dicarboxylic acids. However, oxidation
of galactose yields an insoluble mucic acid. Lactose will also yield a mucic acid, due
to hydrolysis of the glycosidic linkage between its glucose and galactose subunits.
Being insoluble, galactosaccharic acid crystals separate out. In this experiment, it
appears that our Galactose has visible crystals. Whereas, the rest of the solutions had
no visible crystals. (see pictures in the next page).

VI. Mucic Acid Test.