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ANALYSIS OF CARBOHYDRATES
Name:
Group &
Sec:
Bridgette M.
Juarez
Group 3 Sec EF
Date
Performed:
Date
Submitted:
October 24,
2016
November 16,
2016
I.Objective:
1. To identify carbohydrates based on their reactions on different tests
II. Results:
A.General Test for Carbohydrates
Table A1. Molisch Test
Samples
1% Glucose
1% Sucrose
1% Arabinose
1% Starch
Saliva
Pineapple juice
Cotton
Liver filtrate
Tulya Filtrate
Observations
Purple layer in the middle; cloudy layer on top; green
layer under purple layer; transparent layer at the
bottom
Purple layer in the middle; cloudy layer on top; brown
layer under purple layer; transparent layer at the
bottom
Purple layer in the middle (less than sucrose); cloudy
layer on top; brown layer under purple layer
transparent layer at the bottom
Purple layer in the middle; cloudy layer on top (more
cloudy than other samples); green layer under purple
layer; pale green on the bottom
No purple layer; green layer in the middle; transparent
layer with bubbles on top; transparent layer on the on
the bottom
Dark color layer in the middle; yellow in top; brown
layer below dark layer; transparent on the bottom
2 layers: upper is dark violet and the lower is
transparent
Purple layer in the middle; cloudy on top; transparent
layer on the bottom
Dark brownish-violet layer in the middle; murky green
layer on top; brown layer below violet layer;
transparent layer on the bottom
Observations
Extremely dark green solution, almost black
Extremely dark green solution, almost black
Extremely dark green solution, almost black
Extremely dark green solution, almost black
Dark green solution, least dark color solution among
all test samples
Extremely dark green solution, almost black
Yellowish solution with brown particles at the bottom
Liver filtrate
Tulya Filtrate
Extremely dark green solution, almost black
Theoretical Result: deep green color
Experimental result: black coloration
Observations
Dull violet color; most precipitate observed among
the other test solutions
Brighter violet color; few precipitate
Yellow color; few precipitate
Light yellow color; no precipitate
Very light yellow; becomes colorless after 5 minutes
Colorless
Dark yellow; few precipitate
C. Hydrolysis of Polysaccharides
Table C1.
Samples
1% starch
Dextrin
Cotton
Inulin
Liver filtrate
Tulya filtrate
Observations
Colorless solution
Colorless solution
Heterogenous solution
Yellow solution; dark red particles floating on the
entire solution but more accumulated at the
bottom of the test tube
Faint yellow solution; clear solution; no precipitate
Muddy brown colored solution; turbid
2%
fructose
2%
arabinose
2%
Observations
Barfoeds Test
Seliwanoff
Test
th
(4 ) deep blue
Orange
solution; brick
solution
red ppt.
Orcinol test
Cherry red
solution
Greenish
yellow ppt
Clear light
yellow soln;
no sign of
change
Soln turned
brownish
yellow with
ppt
Light blue
solution
Orange
Soln turned
maltose
brown soln;
brighter than
arabinose
(-) light blue
solution; no
significant
change
soln; no ppt
solution
dark violet
with ppt
Cherry red
solution
5%
galactose
Red-orange
soln with ppt.
5%
sucrose
(2nd) deepest
blue soln
among
samples; brick
red ppt
(-) light blue
soln; no ppt.
Soln become
more intense
yellow
compared to
the original
ligter yellow
soln
Solution
turned violet
2%
sucrose
Cherry red
soln
Soltion turned
brownish
yellow
Benedicts Test
Faint yellow color
Observations
Barfoeds Test
faint clear yellow
Dextrin
Cotton
Inulin
Faint yellow;
almost colorless
Murky faint yellow
Tulya
Liver
Seliwanoff Test
Clear yelloworange soln
Clear yelloworange soln
Dark yelloworange soln
Cherry bright red
soln
Blood red soln
Clear yelloworange soln
Benedicts Test
Clear soln; no ppt
Yellow to red pp.
formed
Red ppt at the
bottom
Red ppt at the
bottom
Yellowish-light
brown soln with
ppt
Brwn red ppt
formed
Observations
Barfoeds Test
Soln with red ppt
Light yellow soln
Orcinol Test
Yellow soln
Yellow soln with
brown ppt
Light brown soln
Yellow ppt.
formed
5% sucrose
Starch
Dextrin
Cotton
Inulin
Tulya
Yellow soln
Yellow soln
Green soln
Light blue green
soln
Liver
Yellow soln
III. Discussions:
Carbohydrates originally refer to compounds with the general formula:
Cn (H 2 O)n . But not all sugars fit this formula, only the monosaccharides that
consists of a single polyhydroxy aldehyde or ketone unit. Other sugars such as
oligosaccharides and polysaccharides that are based on the monosaccharide
units have slightly different general formulas. The difference in the general
formulas is caused by the loss of a water molecule for each newly formed link in
non-monosaccharide sugars. Carbohydrates can either be an aldose, the
carbonyl group at the end of its chain is an aldehyde, or a ketose which has a
ketone at the end of its chain. The common number of carbons in each
monosaccharide can range from three to six.
The tests conducted for carbohydrates can be classified into two: general
and specific. The general tests include Molisch test and Anthrone test, while the
specific tests include iodine test, Benedicts test, Barfoed test, Seliwanoff test,
Orcinol test, Osazone test and Mucic acid test. The general tests are used to
detect the presence of sugar in a qualitative or quantitative manner. The specific
tests are used to detect certain sugars or group of sugars according to their
structure or reaction.
The samples for this experiment consist of monosaccharides (glucose,
galactose, fructose & arabinose), oligosaccharides (maltose, dextrin, sucrose &
gum arabic), polysaccharides (cotton, starch), extracts (tulya, liver, saliva and
pineapple juice)
MOLISCH TEST:
This is a qualitative test for presence of carbohydrates. The sample is first
treated with a strong acid (conc.
Arabinose
Glucose
Sucrose
Cotton
Starch
Tulya
Pineapple
Liver
Negative test:
Saliva
ANTHRONE TEST:
The same with Molisch test, Anthrone test is also used to detect the
presence of sugars. The difference with this test from Molisch is that it is more of
a quantitative test that bases the amount of sugar present on the intensity of the
color given in the result. The carbohydrates are first hydrolyzed with sulfuric acid
to form furfurals and/or hydroxy-methly furfural. These furfural are then
condensed by anthrone reagent to form a blue green color complex. (Fig 2)
Fig. 2
Tulya
Negative test:
IODINE TEST
The amylose constituent of starch has helical structure and when it reacts
with iodine solution, the iodine gets trapped inside the helical structure giving
the solution blue color, which indicates the presence of starch. (Fig. 3)
Fig. 3
Starch-iodine complex
I 3
I 2+ I
BENEDICTS TEST
This test is for the presence of reducing sugars that uses an alkaline
reagent. The reagent is composed of copper sulfate that provides cupric ions,
Sodium carbonate which causes the alkalinity of the solution and Sodium citrate
that prevents cupric ions from precipitating. The mixture is then heated in a
water bath. A positive result for the presence of reducing sugars is indicated by
the formation of a precipitate and a change in color. Benedicts reagent contains
blue copper (II) ions, which are reduced to copper (I). These are precipitated as
brick-red copper (I) oxide, which is not soluble in water (Fig. 4). The color of the
solution depends on the concentration of the sugar present (Fig. 4.1)
Fig. 4
Fig. 4.1
All sugars except for sucrose gave a positive result (Fig. 4.2 & Fig. 4.3) since
maltose along with the other monosaccharides are reducing sugars. The
concentration of sucrose did not matter sice its not a reducing sugar. Most of the
monosaccharides reacted fast giving a brick-red precipitate while maltose was in
second to the last. Arabinose was the last to form precipitate probably due to the
fact that the dropping of the Benedicts reagent to the samples were not
simultaneous.
Fig. 4.2 Positive results
Maltose
Arabinose
Glucose
Galactose
Fructose
Sucrose
BARFOEDS TEST
Similar to Benedicts test except for the use of an acidic reagent, Barfoeds
test is also a test for reducing sugars. Monosaccharides are oxidized by the
copper ion in solution to form a carboxylic acid and a reddish precipitate of
copper (I) oxide (Fig. 5). Reducing disaccharides undergo the same reaction, but
do so at a slower rate.
Fig. 5
All the monosaccharides gave a positive result for this test while maltose
and sucrose did not form red precipitates (Fig. 5.1). Maltose is a disaccharide
reducing sugar so it should have tested positive but should be allotted longer
time.
Fig. 5.1
Positive test ex.:
Fructose
Negative test:
Sucrose
Maltose
Sucrose
SELIWANOFFS TEST
This test is used to differentiate ketoses from aldoses. The reagent
dehydrates ketohexoses to form 5-hydroxymethylfurfural. 5hydroxymethylfurfural further reacts with resorcinol present in the test reagent
to produce a red product. Aldohexoses react to form the same product, but do so
more slowly (Fig. 6)
Fig. 6
Since in all the sugars used only fructose is a ketose and there is fructose in
sucrose, theyre the only ones that should give the positive result of having
cherry red solution (Fig. 6.1). Other sugars are more yellow in color than red.
Fig. 6.1 Positive results
Sucrose
Fructose
ORCINOL TEST
This test is specific for pentoses. The components of the reagent include
orcinol, hydrochloric acid, and ferric chloride. When pentose is present, it will be
dehydrated to form furfural which then reacts with the orcinol to generate a
colored substance. The solution will turn bluish and a precipitate may form (Fig.
7).
Fig. 7
Since arabinose is the only pentose sugar in the samples it is the only one
that gave a light blue solution upon its reaction (Fig. 7.1). The other sugars
produced a yellow solution.
Fig. 7.1 Positive test
Arabinose
OSAZONE TEST
Reducing sugars which have either a free aldehyde or keto group are
reacted with phenylhydrazine to form osazone crystals. Three molecules of
phenylhydrazine reacts with the first two carbons in the carbohydrate chain. This
affects the -carbon oxidation with the formation of bis-phenylhydrazone or
osazone (Fig. 8). The crystals formed are either ball-shaped crystals by lactose,
broomstick/needle-shaped crystals from glucose and fructose or sunflowershaped crystals from maltose.
Fig. 8
Galactose
IV. Conclusion
Carbohydrates can be detected by using general tests that can either be
qualitative or quantitative. They can also be identified by using specific tests
that react with sugars with certain characteristics or structures. Usually
monosaccharides are more reactive than oligosaccharides or polysaccharides
and reducing sugars are more reactive than the non-reducing sugars because
they are easily oxidized.
V. References
https://allmedicalstuff.com/anthrone-test-carbohydrates/
http://www.permahealthcare.com/glme-composition.htm
https://books.google.com.ph/books?
id=Ws570Ql8krAC&pg=PA33&lpg=PA33&dq=lab+manual+in+biochemistry+a
nthrone+test&source=bl&ots=ubuZqvTxtG&sig=1g2Dj3ejB6olDV0FWPblEpfTeT
s&hl=en&sa=X&ved=0ahUKEwjwPCigKvQAhXCJ5QKHbHmCvMQ6AEIIDAB#v=onepage&q=lab%20manual%20in
%20biochemistry%20anthrone%20test&f=false
http://eprints.utm.my/6174/1/MasnizaSairi2004_ChemicalCompositionAndSens
oryAnalysis.pdf
https://www.google.com.ph/url?
sa=t&rct=j&q=&esrc=s&source=web&cd=5&cad=rja&uact=8&ved=0ahUKEw
jl0PPp-KrQAhVFs48KHR__A48QFggxMAQ&url=http%3A%2F%2Fwww.umich.edu
%2F~bmsteach%2Flopatin%2Fsalivarygland%2Flectures%2Fdownload
%2FChem_Comp_%26_Funct.ppt&usg=AFQjCNF6o7cWICGhCj1V1rwhEZY_eWBng
http://chemistry.elmhurst.edu/vchembook/548starchiodine.html
https://www.reference.com/science/benedict-test-function-a2b30da3be1056be
http://usmle.biochemistryformedics.com/osazone-test/
https://www.scribd.com/doc/60136819/Osazone-Test
http://generalchemistrylab.blogspot.com/2011/12/mucic-acid-test-forgalactose.html