Introduction
Carbohydrates are one of the most important components in many foods.
Carbohydrates may be present as isolated molecules or they may be
physically associated or chemically bound to other molecules. Individual
molecules can be classified according to the number of monomers that they
contain as monosaccharides, oligosaccharides or polysaccharides.
Monosaccharides
Monosaccharides are water-soluble crystalline compounds. They are
aliphatic aldehydes or ketones which contain one carbonyl group and one or more
hydroxyl groups. Most natural monosachharides have either five (pentoses) or six
(hexoses) carbon atoms.. The reactive centers of monosaccharides are the carbonyl
and hydroxyl groups.
Oligosaccharides
These are relatively low molecular weight polymers of monosaccharides (< 20) that
are covalently bonded through glycosidic linkages. Disaccharides consist of two
monomers, whereas trisaccharides consist of three. Oligosaccharides containing
glucose, fructose and galactose monomers are the most commonly occurring in
foods.
Polysaccharides
The majority of carbohydrates found in nature are present as polysaccharides.
Polysaccharides are high molecular weight polymers of monosaccharides (> 20).
Polysaccharides containing all the same monosaccharides are
called homopolysaccharides (e.g., starch, cellulose and glycogen are formed from
only glucose), whereas those which contain more than one type of monomer are
known asheteropolysaccharides (e.g., pectin, hemicellulose and gums).
Molecules in which the carbohydrates are covalently attached to proteins
are known as glycoproteins, whereas those in which the carbohydrates are
covalently attached to lipids are known as glycolipids. Some carbohydrates
are digestible by humans and therefore provide an important source of
energy, whereas others are indigestible and therefore do not provide energy.
Numerous tests have been devised for the determination of the properties
and for the differentiation of carbohydrates . The main objective of this
experiment is to observe the physical and chemical properties of some
common carbohydrates. Differences in their properties can be significantly
seen through several analytical and experimental tests. This helps greatly in
the characterization of the carbohydrates according to their similar physical
and chemical properties. To be more specific this physical and chemical
properties observed throughout the experiment helps to distinguish between
monosaccharides,disaccharides and polysaccharides.
Results
Benedict’s Test
No Sample Observation Interpretation
1 Glucose Reddish orange colour is It is a reducing
formed sugar
2 Fructose Reddish orange colour is It is a reducing
formed sugar
3 Sucrose No changes. Remains It is a non
blue in colour. -reducing sugar
4 Lactose Reddish orange colour Although it is a
but with a lighter shade disaccharide, it
is formed. is a reducing
sugar
5 Starch No changes . Remains It is non
blue in colour reducing sugar.
6 Water No changes. Remains It acts as a
blue in colour negative
control of the
experiment.
Seliwanoff’s test
No Sample Observation Interpretation
1 Glucose No changes. Ketose is
Remains absent
colourless
2 Fructose From colourless Ketose is
it turns to present
cherry red in a
fast rate(Fast
colour change)
3 Sucrose From colourless Ketose is
it turns to absent
cherry red with
lesser intensity
and slower
rate(Slow
colour change)
4 Lactose No changes. Ketose is
Remains absent
colourless.
5 Starch No changes. Ketose is
Remains absent
colourless.
6 Water No changes. Ketose is
Remains absent. Acts as
colourless. a negative
control of the
experiment
Iodine test
No Sample Observation Interpretation
1 Glucose Light brown Amylose is
colour is absent. It is a
formed monosaccharid
e
2 Fructose Light brown Amylose is
colour is absent. It is a
formed monosaccharid
e
3 Sucrose Dark brown Amylose is
colour is absent. It is
formed disaccharide
4 Lactose Dark brown Amylose is
colour is absent. It is a
formed disaccharide
5 Starch Blue-black Amylose is
solution is present. It is a
formed polysaccharide
6 Water Light yellowish Amylose is
brown solution absent. It acts
is formed as a negative
control of the
experiment
Hydrolysis of Disaccharides and Polysaccahrides
Sucrose+H2 Sucrose +HCl Starch+H20 Starch+HCl
O
Iodine test Light brown Colourless Blue black Colourless
colour solution solution
formed
Benedict’s Light blue Reddish Light blue Very little
test solution orange solution(Negat red
(Negative solution is ive result) precipitate
result) formed. is formed
(Positive
result)
Hydrolysis No Hydrolysis No hydrolysis Starch
product hydrolysis took place. took place might be
present took place Sucrose is hydrolysed
hydrolysed into
into glucose maltose.
and fructose. Hence this
culd be a
false
positive
result
Barfoed’s Test
No Sample Observation Interpretation
1 Glucose Red precipitate is formed in It is a
a faster rate monosaccahrid
e
2 Fructose Red precipitate is formed in It is a
a faster rate monosaccharid
e
3 Sucrose No red precipitate is Not a
formed monosaccharid
e
4 Lactose No red precipitate is Not a
formed monosaccharid
e
5 Starch No red precipitate is Not a
formed monosaccharid
e
6 Water No red precipitate is Not a
formed monosaccharid
e. Acts a
negative
control of the
experiment
Osazone Test
No Sample Observation Interpretation
1 Glucose Broomstick like It is a
shaped crystals glucosazone
are formed
2 Fructose Broomstick like It is a
shaped crystals fructosazone
are formed
3 Lactose Powderpuff like It is a
balls of crystals lactosazone
are formed
4 Maltose Flower shaped It is a
crystals are maltosazone
formed
5 Water No crystals are Acts as a
formed negative
control of the
experiment
Bial’s test
No Sample Observation Interpretation
1 Glucose Yellow colour Not a pentose
solution sugar
2 Fructose Grey colour Not a pentose
solution sugar
3 Sucrose Grey colour Not a pentose
solution sugar
4 Lactose Greyish yellow Not a pentose
colour solution sugar
5 Starch Yellow colour Not a pentose
solution sugar
6 Water Lightest yellow Not a pentose
colour solution sugar. Acts as a
negative
control of the
experiment
Discussion
Post lab questions
When the Cu2+ in Benedict’s solution is reduced , the copper is reduced
from +2 to +1. The glucose and galactose are monosaccharides consisting of
aldehyde group. When the Benedict’s solution is reduced the sugar will be
oxidized where the aldehyde group in the sugar will be oxidized into a
carboxylic acid group.
In Seliwanaoff test, Fructose will form a red color more rapidly since it is a
ketose sugar. Seliwanoff's agent is most sensitive and specific to ketose.
Glucose, which is an aldose will develop slowly giving off a pink color.
Amylose is a polysaccharide made of alpha glucose sugars joined by1-4
glycosidic bonds. Amylopectin is also a polysaccharide made of alpha
glucose sugars joined by1-4 glycosidic bonds but it has 1-6 branches. Both
amylose and amylopectin are of plant origin and are the two components of
starch. Glycogen is the animal equivalent of amylopectin and is found in liver
and muscle. Like starch it is an energy storage compound. In short, Amylose
is a type of starch stored in plant cells, while glycogen is used to store
energy in animal cells. The iodine test is used to see if a substance contains
starch or not.
A reducing sugar is any sugar that is capable of acting as a reducing agent
because it has a free aldehyde group or a free ketone group. All
monosaccharides are reducing sugars, along with some disaccharides,
oligosaccharides, and polysaccharides. All monosaccharides are reducing
sugars because all monosaccharides have an aldehyde group (if they are
aldoses) or can tautomerize in solution to form an aldehyde group (if they
are ketoses).The examples of common monosaccharides are like galactose,
glucose, glyceraldehyde, fructose, ribose, and xylose.Many
disaccharides, like cellobiose, lactose and maltose, also have a reducing
form, as one of the two units may have an open-chain form with an aldehyde
group. Besides that glucose polymers such as starch and starch-derivatives
like glucose syrup, maltodextrin and dextrin are also reducing sugar.
Moreover, glycogen is also another example of reducing sugar but only with
one reducing end.
A non-reducing sugar is a sugar that cannot donate electrons to other
molecules and therefore cannot act as a reducing agent. Sucrose is the
most common nonreducing sugar. The linkage between the glucose and
fructose units in sucrose, which involves aldehyde and ketone groups, is
responsible for the inability of sucrose to act as a reducing sugar. Trehalose
and starch are also another examples of non-reducing sugar
A ketose is a monosaccharide containing one ketone group per
molecule.With three carbon atoms, dihydroxyacetone is the simplest of all
ketoses and is the only one having no optical activity. Ketoses can isomerize
into an aldose when the carbonyl group is located at the end of the molecule.
Such ketoses are reducing sugars. Examples of ketose sugars are fructose ,
ribulose, xylulose and erythrulose .
Benedict's solution contains Copper, which can accept electrons from
reducing sugars and consequently change color. A positive Benedict's sugar
test will produce an orange to brick-red color. Reducing sugars have either a
free aldehyde functional group or a free ketone functional group as part of
their molecular structure; starches and other polysaccharides lack these
functional groups. If Benedict's test changes color ( pos. reaction )
polysaccharides must be hydrolyzed.
Iodine reacts or forms a complex with native unhydrolyzed starch molecule
giving you a characteristic blue color. If starch is completely hydrolyzed, the
color will disappear, Alternatively it will fade to different degrees depending
upon the rate/degree of starch hydrolysis. Hydrolysis of starch were proven
in this experiment because when iodine reagent is added the solution turns
colorless.
Experiment
The copper(II) sulphate is the reduced species in this experiment. The blue
copper (II) ions from copper(II) sulphate are reduced to red copper(I) ions by
the aldehyde groups in the reducing sugars. This accounts for the colour
changes observed. The red copper (I) oxide formed is insoluble in water and
is precipitated out of solution. This accounts for the precipitate formed. As
the concentration of reducing sugar increases, the nearer the final colour is
to brick-red and the greater the precipitate formed.
The principle of the Benedict's Test for non-reducing sugar is when
disaccharides are hydrolyzed to their constituent monosaccharides when
boiled in dilute hydrochloric acid.
The monosaccharide products of hydrolysis are reducing sugars i.e. have the
aldehyde functional group and can reduce copper in the presence of alkali
producing the color changes.
Lactose is a disaccharide but it is a reducing sugar because reducing
disaccharides like lactose and maltose have only one of their two anomeric
carbons involved in the glycosidic bond, meaning that they can convert to an
open-chain form with an aldehyde group. But, nonreducing disaccharides like
sucrose and trehalose have glycosidic bonds between their anomeric carbons
and thus cannot convert to an open-chain form with an aldehyde group; they
are stuck in the cyclic form.
In the Seliwanoff test, which is specific in testing for ketose fructose shows
the positive result . But sucrose has a lighter intensity because the mono
derivatives of sucrose are glucose and fructose. So the reason why sucrose
could have a lighter intensity due to presence of the mono derivative
fructose.
Conclusion
In conclusion there are many vital roles of carbohydrates. The carbohydrates
are a major source of metabolic energy, both for plants and for animals that
depend on plants for food. Aside from the sugars and starches that meet this
vital nutritional role, carbohydrates also serve as a structural material
(cellulose), a component of the energy transport compound ATP, recognition
sites on cell surfaces, and one of three essential components of DNA and
RNA.
It can be concluded that several analytical tests can be done to confirm the
presence of carbohydrates and to distinguish between the classes of
carbohydrates. Different test give different results according to specificity
and sensitivity of the test.
