Name of Experiment: Test for Carbohydrates

Name:

Student ID:

Date of the experiment was performed: 3 November 2022

Module Code: PHC62104

3. Result
1. Molisch’s Test

Table 1: Molisch’s Test for the presence of carbohydrates in glucose, fructose,

sucrose, lactose, starch, and water

No Sample Observation Interpretation

1 Glucose Heat Release out, dark purple Dark purple precipitate

precipitate formed formed in the sample
showing that the presence
of carbohydrates. Glucose
is a type of carbohydrates.
There is positive result.
2 Fructose Heat release out, deep purple Purple solution formed in
solution formed the sample showing that
the presence of
carbohydrates. Fructose is
a type of carbohydrates.
There is positive result.
3 Sucrose Heat release out, deep purple Deep purple solution
solution formed formed in the sample
showing that the presence
of carbohydrates. Sucrose
is a type of carbohydrates.
There is positive result
4 Lactose Heat released out, deep purple colour Deep purple solution
complex form at the bottom of the formed in the sample
solution while the solution at the top showing that the presence
remains milky white of carbohydrates. Lactose
is a type of carbohydrates.
There is positive result.
5 Starch Heat released out, purple ring formed Purple ring formed in the
on the top of the solution. Deep sample showing that the
purple colour complex form at the presence of carbohydrates.
bottom of the solution while the Starch is a type of
 solution at the middle remains milky carbohydrates. There is
white positive result

6 Water Heat release, the solution remain There is no purple solution

yellowish colour formed so there is no
presence of carbohydrates.
Water is not a type of
carbohydrates. There is
negative result.
 2. Benedict’s test for Reducing Sugars

No Sample Observation Interpretation

1 Glucose The solution turns from blue to Glucose is a type of
brick-red precipitate solution; reducing sugar. There is
oxidation reaction has occurred positive result

2 Fructose The solution turns from blue to Fructose is a type of

reddish-orange colour solution; reducing sugar. There is
oxidation reaction has occurred positive result

3 Sucrose The solution remains blue colour. Sucrose is not a type of

Oxidation does not occur reducing sugar. There is
negative result
4 Lactose The solution turns from blue to Lactose is a type of
moderate reddish orange colour; reducing sugar. There is
oxidation reaction has occurred positive result

5 Starch The solution remains blue colour; Starch is not a type of

oxidation does not occurred reducing sugar. There is
negative result

6 Water The solution remains blue colour. Water is not a type of

Oxidation reaction does not occurred reducing sugar. There is
negative result
 3. Seliwanoff’s Test for Ketoses

No Sample Observation Interpretation

1 Glucose There is no changes in the solution. Glucose is not a type of

The solution remains colourless ketose. There is negative
result

2 Fructose The solution changes from colourless Fructose is a type of

to red solution. There is fast changes ketose. There is positive
of the colour of the solution. result

3 Sucrose The solution changes from colourless Sucrose is a type of ketose.

to pale red solution. There is slow There is positive result
changes of the colour of the solution.
4 Lactose There is no changes in the solution. Lactose is not a type of
The solution remains colourless ketose. There is negative
result
5 Starch There is no changes in the solution. Starch is not a type of
The solution remains colourless ketose. There is negative
result
6 Water There is no changes in the solution. Water is not a type of
The solution remains colourless ketose. There is negative
result
 4. Iodine test for polysaccharides

Spot plates labelling Test tube labelling

No Sample Observation Interpretation

1 Glucose There is no changes in the solution. Glucose does not have
The solution remains yellow colour amylose. Thus glucose is
not a type of
polysaccharides. There is
negative result
2 Fructose There is no changes in the solution. Fructose does not have
The solution remains yellow colour amylose. Thus fructose is
not a type of
polysaccharides. There is
negative result
3 Sucrose There is no changes in the solution. Sucrose does not have
The solution remains yellow colour amylose. Thus sucrose is
not a type of
polysaccharides. There is
negative result
4 Lactose There is no changes in the solution. Lactose does not have
The solution remains yellow colour amylose. Thus, lactose is
not a type of
polysaccharides. There is
negative result
5 Starch The solution changes from yellow Starch contains amylose.
colour to dark blue-black colour. Thus, starch is a type of
polysaccharides. There is
positive result
6 Water There is no changes in the solution. Water does not have
The solution remains yellow colour amylose. Thus, water is
not a type of
polysaccharides. There is
negative result.
 5. Hydrolysis of Disaccharides and Polysaccharides

Spot plates labelling Test tube labelling

Sample/ Iodine Test Benedict’s Test Hydrolysis Product

Test present
Sucrose There is no changes in There is no changes in Product of hydrolysis
+ H2O the colour of the solution the colour of solution in does not present. There
in figure 5.1. The figure 5.2. The solution is no changes in both
solution remains remains blue colour iodine test and
brownish-yellow Benedict’s test. That
means no present of
polysaccharide and
reducing sugar in the
solution.
Sucrose There is no changes in The colour of solution Hydrolysis product
+ HCl the colour of the solution changes from blue to present in the solution.
in figure 5.1. The solution that has brick- Sucrose is hydrolysed
solution remains red precipitates. into glucose and
brownish-yellow fructose by HCl. There
is no changes in the
iodine test and this
shows that there is no
presence of
polysaccharides. In
Benedict’s test, brick-
red precipitate present,
reducing sugar present
in the solution
Starch The colour of the There is no changes in Hydrolysis product
+ H2O solution changes from the colour of the solution present in the solution.
brownish-yellow to dark in figure 5.2. The Starch is hydrolysed
blue solution in figure solution remains blue into glucose and
5.1 colour maltose. In iodine test,
dark blue solution
formed and this shows
 that polysaccharides is
present. There are no
changes in Benedict’s
test so there is no
reducing sugar present
in the solution
Starch The colour of the There is no changes in Hydrolysis product
+ HCl solution changes from the colour of the solution present in the solution.
brownish-yellow to dark in figure 5.2. The Starch is hydrolysed
blue solution in figure solution remains blue into simple monomer
5.1 colour of glucose and maltose.
In iodine test, dark blue
solution formed and
this shows that
polysaccharides is
present. There are no
changes in Benedict’s
test so there is no
reducing sugar present
in the solution
H2O There is no changes in There is no changes in Water act as negative
the colour of the solution the colour of solution in control and thus the
in figure 5.1. The figure 5.2. The solution product of hydrolysis
solution remains remains blue colour does not present in
brownish-yellow both iodine and
Benedict’s test
solution.
 6. Barfoed’s Test

No Sample Observation Interpretation

1 Glucose Brick-red cuprous oxide precipitate Glucose is a type of
formed in the bottom of the solution. monosaccharide. There is
The solution at the top remains blue positive result
colour
2 Fructose Brick-red cuprous oxide precipitate Fructose is a type of
formed in the bottom of the solution. monosaccharide. There is
The solution at the top remains blue positive result
colour
3 Sucrose There is no changes in the solution. Sucrose is not a type of
The solution remains blue colour monosaccharides. There is
negative result
4 Lactose There is no changes in the solution. Lactose is not a type of
The solution remains blue colour monosaccharide. There is
negative result
5 Starch There is no changes in the solution. Starch is not a type of
The solution remains blue colour. monosaccharides. There is
negative result
6 Water There is no changes in the solution. Water act as negative
The solution remains blue colour. control in the experiment
thus water is not a
monosaccharide.
7. Osazone test

Osazone Test for the presence of osazone crystals in glucose, fructose, sucrose,
lactose, maltose, and water

Sample Observation Interpretation

Glucose The oxazone The needle-shaped

crystals present in yellow crystals
Figure 7.1 appeared formed, showing that
as needle-shaped glucosazone presents
yellow oxazone in the solution
because glucose is a
type of reducing
sugar

Fructose The oxazone The needle-shaped

crystals present in yellow crystal
Figure 7.2 appeared formed, showing that
as needle-shaped fructosazone
yellow oxazone presents in the
solution because
fructose is a type of
reducing sugar.
Sucrose The oxazone From the beginning,
crystal present in sucrose solution does
Figure 7.3 appeared not form any crystal
as powderpuff but after 30 minutes
yellow osazone boiling, there is
after 30 minutes of powderpuff yellow
boiling crystal formed as the
sucrose is
hydrolyzed into
glucose and fructose
Lactose The oxazone The ball-shaped
crystal observed in yellow oxazone
Figure 7.4 appeared formed showing that
as ball-shaped lactosazone present
yellow osazone in the solution
because lactose is a
type of reducing
sugar

Maltose The oxazone The sunflower-

crystal observed in shaped osazone
Figure 7.5 appeared crystal formed
as sunflower- showing that
shaped osazone maltosazone presents
crystal in the solution
because maltose is a
type of reducing
sugar

Water There is no Water act as negative

oxazone crystal control and thus it
observed in Figure does not form
7.6 osazone
 8. Bial’s Test

No Sample Observation Interpretation

1 Glucose There is no change in the colour of Glucose is a hexose sugar

the solution. The colour of solution
remains yellow

2 Fructose There is no change in the colour of Fructose is a hexose sugar

the solution. The colour of the
solution remains brownish-yellow

3 Sucrose There is no change in the colour of Sucrose is a hexose sugar

the solution. The colour of the
solution remains brownish-yellow
4 Lactose There is no change in the colour of Lactose is a hexose sugar
the solution. The colour of the
solution remains yellow
5 D-ribose Bluish-green solution formed D-ribose is a pentose sugar

6 Water There is no changes in the solution. Water act as negative

The colour of the solution remains control in the experiment
yellow thus water is not a pentose
or hexose sugar
4. Discussion

For the result shown in the first test whichis Molisch’s Test, the water appeared yellow.

Glucose, Fructose, Sucrose, Lactose and starch are seen to either develop a purple ring or

change their colour to purple. It is because concentrated sulfuric acid will dissolve the hydroxyl

group from sugars when they are combined with the Molisch reagent to produce furfural from

pentose sugar and hydroxymethyl furfural from hexose sugar. The existence of a purple result

is caused by the Molisch reagent's reaction of furfural with alpha-naphthol. Thus, glucose,

fructose, sucrose, lactose and starch are type of carbohydrates.

Besides that, in Benedict’s test for reducing sugar, reducing sugar will precipitate as a

brick-red precipitate. Thus, reducing sugar includes glucose, fructose and lactose as oxidation

reaction occurred and the colour of solution changes from blue to brick-red. According to the

research, all monosaccharides are reducing sugar. In their structures, they contain free reducing

ketone or aldehyde. With the exception of sucrose, disaccharides like maltose and lactose are

reducing sugars, as shown in the result in the table above. When the Cu2+ in Benedict's is

reduced , glucose, fructose, or galactose is oxidised. Copper is reduced from +2 to +1 (Aryal,

2018). These sugars' aldehyde oxidised to form a carboxyl group. As a result, the red Cu+

precipitate, which is actually Cu2O formed from the blue Benedict's solution, which contains

Cu2+.

Based on the result in Seliwanoff’s test, fructose and sucrose contain ketose as they

appeared as red solution in the test. Since fructose is a ketose and can dehydrated more quickly

and gives deeper colour, thus fructose will turn red more rapidly when tested with Seliwanoff’s

reagent. Sucrose is a disaccharides which contains both fructose and glucose so thus it will give
a positive result which will turns the colour of solution from colourless to red. The colour of

sucrose turns to pink due to the present of concentrated HCl. Sucrose is then dehydrated into

glucose and fructose when concentrated HCl react with ketose in sucrose. Then , the fructose

from sucrose react to produce pink solution. Glucose is an aldose so there will be no change

in the colour of glucose and the solution remains clear. However, the colour of glucose solution

will be change after longer period of time because glucose is an aldose and aldose react slowly

and the colour appeared is faint. Based on the result above, ketose dehydrated more rapidly

than aldose. Sucrose react slower as sucrose need to be dehydrated and it is not pure fructose.

The intensity of fructose is higher than sucrose.

In addition, from the iodine test for polysaccharides, some polysaccharides, such as

amylose, dextrin, and glycogen, can be distinguished from other carbohydrates by testing with

iodine test. Iodine tests cannot be used to evaluate branched polysaccharides like cellulose, and

they will remain colourless (Sapkota, 2020). Branched polysaccharides like cellulose,

monosaccharides, and disaccharides remain colourless. An orange-yellow colour is produced

by amylopectin. As can be observed from the result, amylose will generate a dark blue colour,

indicating that starch was hydrolyzed, whereas glycogen will produce a reddish brown colour.

When the environment was acidic, sucrose and starch were hydrolyzed. The monomer

units' glycosidic bonds are disintegrating. Hydrolysis produces monosaccharides from

disaccharides and polysaccharides. In order to make sure that the starch has been converted

into monosaccharides through hydrolysis and do not formed blue solution, an iodine test is

required. Since reducing sugars are formed, Benedict's test indicates this. Since sucrose

hydrolyses to make glucose and fructose, it will produce red precipitate, which will indicate a

positive Benedict's test result. A polysaccharide, starch contains many glycosidic linkages. As
a result, hydrolysis takes longer, and the resulting colour is paler than that of hydrolyzed

sucrose.

In the Barfoed test, glucose and fructose demonstrated that brick-red cuprous oxide

precipitation was appearing while other substances remains unchanged. In the Barfoed test,

monosaccharides are oxidised using glacial acetic acid, a weak acid that reduces Cu2+ to Cu+

more quickly. With this reagent, disaccharides may also react, but the process proceeds

considerably more slowly than it does with monosaccharides.

Practically speaking, the Osazone test observation is distinct from other tests. Under a

microscope, we must check the osazone shapes and structures for each sugar. Additionally,

each osazone crystal will show up in the boiling water bath at its own specific time. In glucose

and fructose, needle-shaped crystals form; in lactose, ball-shaped crystals form; and in maltose,

sunflower-shaped crystals form which indicated that glucose is reducing sugar. Lactose is type

of reducing sugar as the structure and shape of the osazone crystal appeared as ball-shape. In

the osazone test, we can distinguish between various sugars by looking at their shapes. Due to

the need to be hydrolyzed into the reducing sugars which are fructose and glucose during a

lengthy 30-minute boil, sucrose will also eventually crystallise as a powderpuff after the

lengthy boiling period.

In contrast to the other solutions, which are either brown or yellow, our solution is

bluish-green, according to the results of Bial's test. When a pentose sugar is present, the colour

will be blue-green. D-ribose is a pentose as a result. When heated in an acidic medium, D-

ribose dehydrates to appear as furfural, which is blue-green in colour. Ferric chloride and

orcinol with concentrated hydrochloric acid make up the acidic medium that was previously
discussed (asus, 2013). The solution changes colour from yellow to grey or brown when heated

with hexose sugars, whereupon hydroxymethylfurfurals are formed. Glucoronates may also

produce a blue-green precipitate when heated for an extended period of time. Controlling the

time restriction is crucial because doing so can provide a false-positive outcome.

5. Conclusion

In a nutshell, carbs are necessary for us and our daily lives since they give us energy.

We can distinguish between monosaccharides, disaccharides, and polysaccharides through

various tests and evaluate their physical and even chemical features. In summary, the Molisch

test can detect all types of carbohydrates, while Benedict's test is used to identify reducing

sugars, the Iodine test is used to determine the presence of starch and some polysaccharides,

Seliwanoff's test detects ketose, which is composed of fructose and sucrose, Barfoed's test only

detects monosaccharides, Bial's test distinguishes between pentose and hexose

6. Self-reflection

I have learned how to distinguish between different types of carbohydrates, such as

monosaccharides and disaccharides, using a variety of physical and chemical tests, such as

Molisch's test and Seliwanoff's test. Additionally, you gained observation skills and about the

molecular and physical characteristics of carbohydrates. One of the difficulties I had was using

a test tube holder to manage the test tubes with samples and reagents inside of them; it was

difficult to pick up and hold the test tubes because they were almost completely submerged in

the water bath of the beaker, and I had to be careful not to spill the solution or water bath while
holding the test tubes. I must admit that this practical session was both enjoyable and time-

consuming, but it was also rewarding to see the results of our testing at its conclusion. In my

upcoming practical sessions and in my professional pharmacy practice, I'll begin applying the

lessons I've learned.

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