BIOORGANIC

BIOORGANIC
CHEMISTRY
BioOrganic Chemistry CH 205 (2010-2011) Experiment #6

Determination of an unknown carbohydrate

based on the results of different chemical tests
Pamela S. de Leon, Manette C. Dejelo*, James Benedict L. Dolojan and Erik Joseph A. Gatdula
Department of Sports Science, College of Rehabilitation Sciences
University of Santo Tomas, Espaa Street, Manila 108
Date Submitted: October 1, 2010
Abstract:
Carbohydrates are the most abundant organic compounds in the plant world. They act as storehouses of
chemical energy (glucose, starch, glycogen); are components of supportive structures in plants (cellulose),
crustacean shells (chitin), and connective tissues in animals (glucosaminoglycans); and are essential components of
nucleic acids (D-ribose and 2-deoxy-D-ribose). Monosaccharides, oligosaccharides, and polysaccharides are the
classifications of carbohydrates. The name carbohydrate arises from the basic molecular formula (CH2O)n where
n=3 or more. (CH2O)n can be rewritten (C*H2O)n to show that these substances are hydrates of carbon.
This experiment involves the following carbohydrates: sucrose, maltose, glucose, fructose, and xylose. The
unknown sample was to be determined with the use of the Barfoeds test, Benedicts test, Seliwanoffs test, and
Bials Orcinal test. With the results of these tests, we are able to determine to what the unknown sample is; even
though the results are not exact, they are accurate.
Keywords: Carbohydrates, monosaccharides, oligosaccharides, polysaccharides, sucrose, maltose, glucose,
fructose, xylose, Barfoeds test, Benedicts test, Seliwanoffs test, Bials Orcinal test

1. Introduction
Carbohydrates, the most abundant biomolecules in nature, are a direct link between
solar energy and the chemical bond energy of living organisms. More than half of all organic
carbon is found in carbohydrates. They are formed during photosynthesis, a biochemical
process in which light energy is captured and used to drive the biosynthesis of energy-rich

organic molecules from the energy-poor inorganic molecules CO 2 and H2O. Most
carbohydrates contain carbon, hydrogen, and oxygen in the ratio (CH 2O)n, hence the name
hydrate carbon. They have been adapted for a wide variety of biological functions, which
include energy sources, structural elements, cellular communication and identity, and
precursors in the production of other biomolecules. Certain carbohydrate molecules are
structural elements of nucleotides and nucleic acids.
Carbohydrates are classified as monosaccharides, disaccharides, oligosaccharides,
and polysaccharides according to the number of simple sugar units they contain. In the
experiment, the carbohydrates involved are sucrose, maltose, glucose, fructose, xylose.
Sucrose, common table sugar, is the most abundant of all disaccharides and occurs
throughout the plant kingdom. It is produced commercially from the juice of sugar cane and
sugar beets. Maltose, often called malt sugar, is produced whenever the polysaccharide starch
breaks down, as happens in plants when seeds germinate and in human beings during starch
digestion. It is a common ingredient in baby foods and is found in malted milk. Glucose is
the most abundant monosaccharide in nature and the most important from a human
nutritional standpoint. It is originally called dextrose, and it is the primary fuel for living
cells. Fructose is biochemically the most important ketohexose. It is also known as luvolose
and fruit sugar. The sweetest-tasting of all sugars, fructose is found in many fruits and is
present in honey in equal amounts with glucose. Xylose, also called wood sugar, is a pentose
sugar found in plant tissues; 40% as sweet as sucrose.
In the experiment, the tests involved are the Barfoeds test, Benedicts test,
Seliwanoffs test, and Bials Orcinal test. Barfoeds test shows positive tests for reducing
monosaccharides. Benedicts test is for reducing sugars such as glucose, sucrose, and
fructose. Seliwanoffs test shows positive tests for ketoses. Bials test shows positive tests for
pentoses.
In this experiment, the group should be able to achieve the following objectives: (1)
to describe the structural features common to all carbohydrates, (2) to give specific chemical
tests to characterize a given carbohydrate, (3) to identify an unknown carbohydrate based on

the results of the different chemical tests and in comparison with standard carbohydrate, and
(4) to provide the underlying principle involved in carbon chemical test.
2. Methodology
A. Preparation of the solution of the unknown sugar
1g of unknown sugar was given, and then added to10 mL of water. One-fourth of this
was used as the sample for the tests which are the Barfoeds test, Benedicts test,
Seliwanoffs test, and Bials Test.
B. Tests
Barfoeds Test
10 drops of test compounds were placed in separate test tubes. 20 drops of Barfoeds
reagent were then added. Test tubes were placed in boiling water bath for 10 minutes. The
color of each test tube was recorded and the time of the formation of precipitate was noted.
The color of the reagent (blue) was compared.
Benedicts Test
10 drops of test compounds were placed in separate test tubes. 20 drops of Benedicts
reagent were then added. Test tubes were placed in boiling water bath for 5 minutes. The
color of each test tube was recorded and the time of the formation of precipitate was noted.
The color of the reagent (blue) was compared.
Seliwanoffs Test
10 drops of test compounds were placed in separate test tubes. 20 drops of
Seliwanoffs reagent were then added. Test tubes were placed in a boiling water bath for 10
minutes. Test tubes were removed from the water bath when there change in color. The color
of each test tube and the time of its formation were recorded. The color of the reagent
(colorless) was compared.

Bials-Orcinol Test
10 drops of test compounds were placed in separate test tubes. 20 drops of Bials
reagent were then added. Test tubes were placed in a boiling water bath for 10 minutes. Test
tubes were removed from the water bath when there change in color. The color of each test
tube and the time of its formation were recorded. The color of the reagent (yellow) was
compared.
3. Results and Discussion
BARFOEDS
*Blue
Unknown

Sucrose

Maltose

Glucose

Fructose

Xylose

BENEDICTS
*Blue

Clear blue soln

(09:00) redbrown ppt;
clear blue soln
Clear blue soln
Clear blue soln
(no change)

Clear blue soln

(09:40) redbrown ppt;
clear blue soln
Clear blue soln
(09:17) redbrown ppt;
clear blue soln
Clear blue soln
(07:35) redbrown ppt;
clear blue soln
Clear blue soln
(07:30) redbrown ppt;
clear blue soln

SELIWANOFFS
*Colorless

BIALS ORCINOL
*Yellow

Clear blue soln

(03:20) orange
ppt; cloudy
orange soln
Clear blue soln
Clear blue soln
(no change)

Colorless soln
(03:54) clear peach
soln

Clear yellow soln

(04:40) moss green
soln

Colorless soln
(02:13) cloudy
pink soln

Clear yellow soln

(02:56) dark
greenish-brown soln

Clear blue soln

(02:42) orange
ppt; orange soln

Colorless soln
(05:11) clear peach
soln

Clear yellow soln

(05:15) dark green
soln

Clear blue soln

(03:30) orange
ppt; cloudy
orange soln
Clear blue soln
(00:41) orange
ppt; orange soln

Colorless soln
(03:54) clear peach
soln

Clear yellow soln

(05:20) moss green
soln

Colorless soln
(01:46) cloudy
dark pink/red soln

Orange soln
(02:27) brown-black
soln

Clear blue soln

(00:22) orange
ppt (powderlike); orange
soln

Colorless soln
(04:56) black soln

Clear yellow soln

(04:22) dark blue
green soln

o - Before
- After
Unknown sample: GLUCOSE

In Barfoeds test, all of the samples were originally a clear blue solution. The unknown
sample formed a red-brown precipitate in 9 minutes with a clear blue solution. The sucrose
sample had no changes; it remained to be a clear blue solution. The maltose sample formed a
red-brown precipitate in 9 minutes 40 seconds with a clear blue solution. The glucose sample
formed a red-brown precipitate in 9 minutes 17 seconds with a clear blue solution. The fructose
sample formed a red-brown precipitate in 7 minutes 35 seconds with a clear blue solution. The
xylose sample formed a red-brown precipitate in 7 minutes 30 seconds with a clear blue solution.
In Benedicts test, all of the samples were originally a clear blue solution. The unknown
sample formed an orange precipitate in 3 minutes 20 seconds with a cloudy orange solution. The
sucrose sample had no changes; it remained a clear blue solution. The maltose sample formed an
orange precipitate in 2 minutes 42 seconds with an orange solution. The glucose sample formed
an orange precipitate in 3 minutes 30 seconds with a cloudy orange solution. The fructose sample
formed an orange precipitate in 41 seconds with an orange solution. The xylose sample formed
an orange precipitate (powder-like) with an orange solution.
In Seliwanoffs test, all of the samples were originally a colorless solution. The unknown
sample formed a clear peach solution in 3 minutes 54 seconds. The sucrose sample formed a
cloudy pink solution in 2 minutes 13 seconds. The maltose sample formed a clear peach solution
in 5 minutes 11 seconds. The glucose sample formed a clear peach solution in 3 minutes 54
seconds. The fructose sample formed a cloudy dark pink/red solution in 1 minute 46 seconds.
The xylose sample formed a black solution in 4 minutes 56 seconds.
In Bials test, all of the samples were originally a yellow solution except fructose with an
orange solution. The unknown sample formed a moss green solution in 4 minutes 40 seconds.
The sucrose sample formed a dark greenish brown solution in 2 minutes 56 seconds. The
maltose sample formed a dark green solution in 5 minutes 15 seconds. The glucose sample
formed a moss green solution in 5 minutes 20 seconds. The fructose sample formed a brownblack solution in 2 minutes 27 seconds. The xylose sample formed a dark blue-green solution in 4
minutes 22 seconds.

With the help of the results, the unknown sample was determined. The tests show that the
unknown sample is glucose. Going back to the results, in the Barfoeds test, the unknown sample
formed a red-brown precipitate in 9 minutes with a clear blue solution. The glucose sample
formed a red-brown precipitate in 9 minutes 17 seconds with a clear blue solution. In the
Benedicts test, the unknown sample formed an orange precipitate in 3 minutes 20 seconds with a
cloudy orange solution. The glucose sample formed an orange precipitate in 3 minutes 30
seconds with a cloudy orange solution. In the Seliwanoffs test, the unknown sample and the
glucose sample formed a clear peach solution in 3 minutes 54 seconds. In the Bials test, the
unknown sample formed a moss green solution in 4 minutes 40 seconds. The glucose sample
formed a moss green solution in 5 minutes 20 seconds. These results are accurate enough to say
that the unknown sample is glucose.

4. Conclusion
Carbohydrates are the most abundant class of bioorganic molecules on planet Earth.
Although their abundance in the human body is relatively low, carbohydrates constitute about
75% by mass of dry plant materials. Green (chlorophyll-containing) plants produce
carbohydrates via photosynthesis. In this process, carbon dioxide from the air and water from the
soil are the reactants, and sunlight absorbed by chlorophyll is the energy source.
CO2 + H2O + solar energy carbohydrates + O2

Plants have two main uses for the carbohydrates they produce. In the form of cellulose,
carbohydrates serve as structural elements, and in the form of starch, they provide energy
reserves for plants.
A carbohydrate is a polyhydroxy aldehyde, a polyhydroxy ketone, or a compound that
yields polyhydroxy aldehydes or polyhydroxy ketones upon hydrolysis. The carbohydrate
glucose is a polyhydroxy aldehyde, and the carbohydrate fructose is a polyhydroxy ketone. A
monosaccharide is a carbohydrate that contains a single polyhydroxy aldehyde or polyhydroxy

ketone unit. Monosaccharides cannot be broken down into simpler units by hydrolysis reactions.
Pure monosaccharides are water-soluble, white, crystalline solids. Oligosaccharide is a
carbohydrate that contains two to ten monosaccharide units covalently bonded to each other.
Disaccharides are the most common type of oligosaccharide; it is a carbohydrate that contains
two monosaccharide units covalently bonded to each other. Like monosaccharides, disaccharides
are crystalline, water-soluble substances.

Sucrose

Maltose

Glucose

Fructose

Xylose
The unknown sample in the experiment is glucose. D-glucose, originally called dextrose,
is found in large quantities throughout the living world. In animals, glucose is the preferred
energy source of brain cells and cells that have few or no mitochondria, such as erythrocytes.

Cells that have limited oxygen supply, such as those in the eyeball, also use large amounts of
glucose to generate energy. Dietary sources include plant starch and the disaccharides lactose,
maltose, and sucrose. Glucose is one of the products of photosynthesis in plants and some
prokaryotes. In animals and fungi, glucose is the result of the breakdown of glycogen, a process
known as glycogenolysis. In plants, the breakdown substrate is starch. In animals, glucose is
synthesized in the liver and kidneys from non-carbohydrate intermediates, such as pyruvate and
glycerol, by a process known as gluconeogenesis. Glucose, when undergoes the Barfoeds test,
forms a red-brown precipitate with a clear blue solution. When it undergoes the Benedicts test, it
forms an orange precipitate with a cloudy orange solution. When it undergoes the Seliwanoffs
test, it forms a clear peach solution. When it undergoes the Bials test, it produces a moss green
solution.

Glucose (Fischer Projection)

5. Acknowledgements
I would like to thank the following persons:
Mrs. Gelvero and Mr. Layco, our lab instructors in Chemistry Lab 205, who guided us in
conducting the experiment.
The librarians of the Medicine Library and the Miguel de Benavides Library for helping
me find the sources I need to support my results in the experiment.

My fellow group mates, Pamela S. De Leon, James Benedict L. Dolojan, and Erik Joseph
A. Gatdula, for being dedicated to the experiment and showing unity, cooperation, and team
work.

6. References
Stoker, S. (2010). Exploring general, organic, and biological chemistry.
Philippines: ESP Printers Inc.
Foote, C., Iverson, B., & Ansyln, E. (2008). Organic chemistry. (5th ed.).
USA: Brooks/Cole Pub Co.
Garrett, R., Grisham, C. (2009). Biochemistry. (4h ed.).
USA: Chelsea House Publications

i*Corresponding author (Dejelo, Manette C.)