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ABSTRACT
Carbohydrates are defined as any of agroup of organic compounds that
includes sugars, starches, celluloses, and gums and serves as a majorsource of
energy. In this experiment, starch was extracted from a potato and then general
tests for polysaccharides, qualitative tests for carbohydrates, and enzymatic
hydrolysis was performed.
The positive result for Molischs test confirmed the
presence of a carbohydrate. The iodine test is used to test the presence of starch.
Benedicts test was used to detect reducing sugars, Barfoeds test for the presence
of monosaccharides, Seliwanoffstest for ketohexoses, and Bials test for pentoses.
Thin-Layer Chromatography and quantitative analysis was done afterwards.
INTRODUCTION
Carbohydrates are one of the
four essential macromolecules in the
process of life. They serve as fuel in
energy
production.
Carbohydrates
have a general formula of (CH2O) and
are grouped according to the number
of molecules they have.
Monosaccharides are sugars
that cannot be hydrolyzed. They are
further classified by the number of
carbons they contain. Pentoses have
five carbons while hexoses have
six.They can also beclassified as
aldoses or ketoses. Aldoses contain
onealdehyde group while ketoses
contain one ketone groupwithin the
molecule.
Oligosaccharides on the other
hand,
contain
two
to
ten
monosaccharide
units.
They
are
subdivided further according to how
many monosaccharide units they are
made up of.
Polysaccharides
can
becomposed of hundreds of thousands
of monosaccharide units connected
invarious patterns. Common examples
of
polysaccharides
are
starch,
cellulose and glycogen.
Starch, which was isolated in
thisexperiment, is a polymer of
glucose. It is commonly found in
roots, seeds, stems,tubers and corms
of plants, as microscopic granules.
Starch is also essential for animals
and mans nourishment.
Methodology
10
mL
of
the
isolated
carbohydrate was placed in a beaker
and 2.3 mL of saliva was added
afterwards. It was then allowed to
stand at room temperature for 30
minutes. Any change in viscosity of
the solution was noted. A dialyzing
bag was supposed to be used next but
due to the unavailability of this item, a
clear plastic bag was used in place.
The bag was suspended overnight in a
small flask with 50 mL distilled water.
D.
Qualitative
Carbohydrates
Tests
for
1.
Benedicts,
Barfoeds
and
Seliwanoffs
and
Bials-Orcinol
Test
In separate test tubes, five
drops of each of the carbohydrate
sample solutions 0.1M glucose,
fructose, lactose, sucrose and the
hyrdolysate were mixed with 1 mL
of Benedict's reagent. (One test on
the different carbohydrate solutions
must be performed at the same time.)
The test tubes were then placed into a
boiling water bath and were removed
whenever the solutions for one test
gave a visible result. The result and
the time it took for the visible result to
Isolate:
STARCH
Powdery
substance
3. Phenylhydrazone Test
Carbohy
drate
solution
Visible Results
Seliw
Benedi Barfoe
anoff
cts
ds
s
Bial
s
Glucose
Brick
red ppt.
Brick
red ppt.
Orange
soln,
no ppt.
Dark
Green
soln
Fructose
Brick
red ppt.
Brick
red ppt.
Cherry
red
ppt.
--
Xylose
--
--
--
Bluegreen
soln
Lactose
Brick
red
soln
with
brick
red ppt.
No
reaction
Orange
soln,
no ppt
--
Sucrose
Brick
red ppt.
Brick
red ppt.
Cherryred
ppt.
--
Hydrolys
ate:
Starch
Bluegreen
soln
Cherry
red
soln
Yellow
soln
--
Molischs
Solution
solidifie
d with a
blue
layer on
top
Description
Purple
interphas
e in
bottom
layer.
*S = Simple, C= Complex, M=
Monosaccharide, D= Disaccharide, O=
Oligosaccharide,
P=
Polysaccharide, R= Reducing,
NR= Non-reducing, PN= Pentose, HX=
Hexose,
A= Aldose,
K= Ketose
Thin-Layer Chromatography
The
phenylhydrazone
test
detects
reducing
sugars
like
monosaccharides and disaccharides.
Phenylhydrazone reagent consists of
phenylhydrazine hydrochloride and
NaCH3COO. Aldoses and ketoses when
they react to phenyl hydrazine, they
form crystalline product which is
known as the (osazones).
CONCLUSIONS
Sugar
*S
or C
Hydrolysat
e
Standards
Glucose
*M,
O or
P
M
*R
or
NR
R
*PN
or
HX
HX
Fructose
HX
Xylose
PN
Lactose
HX
Sucrose
NR
HX
Starch
NR
HX
Hydrolysate
HX
Distance
traveled
by the
solvent
Distance
traveled
by the
solute
Rf Value
*A Identity
or Kof
compon
ents
Dex
-trin
Malt
-ose
Glucose
Enzyme
7.7
cm
7.7
cm
7.7 cm
7.7 cm
1.4
cm
0.7
cm
1.3 cm
0.2 cm
0.18
cm
0.09
cm
0.17 cm
0.03 cm
--
--
--
Maltose
Quantitative Analysis
4
Absorbance
0
0