NAME: MOHD SAFWAN BIN MOHD SHAFIE

CLASS:M04N
PRACTICAL 5
Title
Objective
Apparatus
Materials
Procedure

: Identification of some biological important chemical compound

: a) To identify some biological important chemical compound
b) To know the specific reagent for certain chemical constituent
: Refer to Biology handbook page 112
: Refer to Biology handbook page 112
: Refer to Biology handbook from page 113 to page 115

Data collection:
PART 1
A.STARCH
Reagent

Substances

Observation

Details

Dilute
iodine

Starch suspension

The solutions colour

changes from yellow to
blue-black.
Potato tuber were
coloured to blue-black
Under microscope
observation, all cells
were coloured into
blue-black

When starch is mixed

with iodine in water,
an intensely colored
starch/iodine complex
is formed. But it seems
that the iodine gets
stuck in the soluble
starch. The starch
forces
the
iodine
atoms into a linear
arrangement in the
central groove of the
soluble starch coil,
giving the complex its
intense blue color.

Potato Tuber

B.GLUCOSE
Reagent
Benedicts
reagent

Subtances
Glucose solution

Observation
The solutions colour
changes from clear
blue to cloudy green to
yellow and finally to a
red precipitate when
boiled.

Details
Aqueous glucose is
mixed with Benedict's
reagent, a solution of
copper sulfate, sodium
hydroxide, and tartaric
acid. The mixture is
heated. Carbohydrates
which
react
with
Benedict's reagent to
reduce the blue copper
(II) ion to form a brick
red
precipitate
of
copper (I) oxide are
classified as reducing
sugars.

C. FRUCTOSE
Reagent
Benedicts
reagent

Subtances
Apple

Observation
Cell change colour from
blue-black to brown
The microscope
observation is attached at
the back.

Details
Aqueous fructose is mixed
with Benedict's reagent, a
solution of copper sulfate,
sodium hydroxide, and
tartaric acid. The mixture is
heated. Carbohydrates
which react with Benedict's
reagent to reduce the blue
copper (II) ion to form a
brick red precipitate of
copper (I) oxide are
classified as reducing
sugars.

D. SUCROSE
Reagent
Benedicts
reagent

Subtances
Sucrose solution

Observation
With no hydrolysing

No colour change
With hydrolysing

The solutions
colour changes
from blue to green
to yellow and
finally to brick-red

Details
Sucrose is disaccharide
that does not reduce
copper
sulphate.
Therefore, it should be
hydrolysed
into
its
constituent
monosaccharides before
testing it with benedicts
reagent. Sucrose then
will react with Benedict's
reagent to reduce the
blue copper (II) ion to
form a brick red
precipitate of copper (I)
oxide.

E. CELLULOSE
Reagent
Schultz
solution

Subtances
Cotton wool

Observation
Cotton is high of
cellulose. Therefore it
should be stained to
purple. However, the
changes of colour of
cotton are not clear.
When the solutions
tested on a plant tissue
and observed under
microscope cell walls
of the tissue is purple in
colour

Details
Schultz solution
reacted with cellulose
to produced purple
stained. In the
experiment, when the
reagent is put on the
cotton and also in the
solution, both of the
produce purplecoloured stains
indicating that both of
them possess cellulose.

Observation
When tested on
toothpick, colour of
toothpick becomes red
When solution is
tested on plants item,
colour of xilem seen
to be red in colour

Details
Acidified
phloroglucinol can
react with lignin to
produce red stain. After
testing with toothpick
and xilem, both of them
stained red indicating
that both of them
contain lignin.

F. LIGNIN
Reagent
Acidified
phloroglucinol

Subtances
Wood

G. LIPIDS
Reagent

Substances

Sudan

Oil palm
and water

Sudan

Pure oil
and palm
oil

Ethanol

Palm oil
and water

Observation
Colour (initial)
Colour (final)
Two separate Two layers are
layers are
formed
formed
Top red and
The top one
bottom dark red
yellow (oil)
the bottom
colourless
(water)
The oil palm The red settles
is pale
at the base of
yellow in
the test tube,
colour
whilst the top
The entire
layer is a dark
yellow with red
solution is
droplets
red in colour
suspended in it
Initially clear, A thin top, layer
slightly
of cloudy, light
cloudy when
yellow,
water is added
followed by a
long rung of
cloudy white
and a thin
bottom layer of
dear yellow

Details
Basically, when Sudan
III reacted with lipid, a
red-stained oil layer
separate out and floats
on the water surface.

Ethanol reacted with

lipids to form a cloudy
white precipitate. In the
experiments, cloudy
white precipitate
formed after some time
representing that the
palm oil contained
lipid.

H. PROTEIN
Reagent
Millons
reagent

Substances
Albumen

Observation
Details
Brick-red precipitate appears. Millon tests for
phenolic -OH group
Pea
Before heated, the sample present in sample such
was creamy white in colour. as protein. Millons
However, after heated, the reagent is prepared
colour of the sample change from mercuric and
mercurous nitrate in
to yellowish browning.
nitric acid. When
protein is heated with
Millon's reagent, a
white precipitate is
formed and on heating
changes to brick red.
Xanthoproteic Nitric
The solutions colour which
Xanthoproteic Acid
test
Test tests R Group.
acid
is deep-yellow disappears.
This is a test for
specific functional
groups that a positive
test is formation of a
deep-yellow orange or
orange-red color. A
deep-yellow partial
precipitate, which
disappears, on shaking,
indicates protein.
Biuret test
Potassium A blue ring appears at the
Biuret array is a test for
peptide bonds present
hydroxide
surface of the solution.
in the sample such as
On shaking, the blue ring
protein. Under alkaline
disappears.
conditions, peptide
The solutions colour turns
bonds are cleaved and
purple
react with a copper
reagent to yield a
purple colour.

I. INVESTIGATING UNKNOWN
Reagent
Dilute iodine

Benedict
reagent

Schultzs
solution

Component
Content

Observation
No colour changes
spotted. The solution
remains yellow
No colour changes
spotted. The solution
remains yellow
No colour changes
spotted. The solution
remains yellow

Husk

Seeds

Content

The solutions colour

changes from blue to
green to yellow to
brick-red

Husk

Seeds

The solutions colour

changes from blue to
green to yellow to
brick-red
The solutions colour
changes from blue to
green to yellow to
brick-red

Content

Husk

Seeds

Content

Husk

Acidified

phloroglucinol

The solutions colour

changes from
colourless to purple
No colour changes
spotted. The solution
remains colourless.
No colour changes
spotted. The solution
remains colourless.
No colour changes
spotted. The solution
colour remains red.
No colour changes

Details
Basically, if starch is
mixed with iodine in
water, an intensely colored
starch/iodine complex is
formed. However, when
iodine is dropped into the
unknown solution, no
colour changes which is
from yellow to blue-black
is spotted. Therefore, there
is no starch in the
unknown.
When benedict reagent is
dropped into the unknown
solution, the colour
changes. This mean the
unknown do has
glucose/carbohydrates in
it. This is because
Carbohydrates react with
Benedict's reagent to
reduce the blue copper (II)
ion to form a brick red
precipitate of copper (I)
oxide. Even so, the
unknown solution does not
have sucrose in it because
red-brick precipitate can
be obtained without
hydrolysing it.
Cellulose stained purple
with Schultz solution.
Then, when the reagent is
being tested with the
unknown solution, the
solutions colour changes
from colourless to purple.
For this reason, the
unknown should comprise
cellulose in it.
Acidified phloroglucinol
can react with lignin to
produce red stain.
However, not all structure

Sudan III

Millons
reagent

Seeds

Content

Husk

Seeds

Content

Husk

Seeds

spotted. The solution

colour remains red.
The solution changes
colour from red to
colourless.
The solution changes
colour from paint
yellow to blurred
red.
No colour changes
spotted. The solution
remains paint yellow
No colour changes
spotted. The solution
remains paint yellow
The solutions colour
changes from
colourless to blurred
brick-red
No colour changes
spotted. The solution
remains colourless.
The solutions colour
changes from
colourless to blurred
brick-red

in the unknown actually

reacted with the reagent to
produce red stained. Only
certain part such as content
and husk stained red with
the reagent
Fundamentally, a redstained oil layer that
separate out and float on
the water surface indicates
the existence of lipid in the
solution. Still, when Sudan
III is added up to the
solution, a blurred redstained oil layer appears.
This mean, there is not
much lipid exist in the
unknown.
Hypothetically, when
protein is heated with
Millon's reagent, a white
precipitate is formed and
on heating changes to
brick red. However, when
the unknown is heated, a
blurred brick-red
precipitate produced,
meaning that the amount
of protein in the unknown
is not much.

PART 2

Chemical
compound

Reagent

Fructose

Benedicts
reagent

Lignin

Sudan III

Protein

Millons
reagent

Observation
Title: Light photomicrograph drawing of cell of apple

Magnification: x100
Title: Light photomicrograph drawing of xilem of
plant stem

Magnification: x100
Title: Light photomicrograph drawing of cell of pea

Magnification: x100

Unknown

Benedicts
reagent

Sudan III

Millons
reagent

Title: Light photomicrograph drawing of unknown cell

Magnification: x100
Title: Light photomicrograph drawing of unknown cell

Magnification: x100
Title: Light photomicrograph drawing of unknown cell

Magnification: x100

PART 3
Conclusion
1.
2.
3.
4.
5.
6.
7.
8.

Potato tuber contained starch because the dilute iodine change colour from yellow to
blue-black
Apple contained glucose and sucrose when brick red precipitates appear when tested
with benedicts reagent
Apple contained fructose because the cells change colour from blue to brown after
heated
Stem cells containing cellulose since the cell stained in purple.
Stem cells contained lignin since the cell stained red in colour.
Oil palm seeds contained lipids
Albumen and pea contained lipid because brick red precipitate appears when tested
with Millons reagent.
For the unknown test, the unknown contained high concentration of fructose, glucose
and sucrose because brick red precipitates appear when tested with Benedicts
reagent. However, apple contained low quantity of cellulose and lignin since the
change of colour of the indicators are blurred. Apple also contained low quantity of
lipid and protein.

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