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A Lab Report on

Analysis of Carbohydrates

CHEM 153L

BSN - C

Submitted by:

Marbella, Mariano

Mendez, Jaira

Mendez, Nessah

Submitted to:

Ms. Faith Galgo


Introduction

Carbohydrates are a standout amongst the most vital components in numerous foods.
carbohydrates might be available as detached atoms or they might be physically related or
chemically bound to different particles. individual atoms can be ordered by the quantity of
monomers that they contain as monosaccharides, oligosaccharides or polysaccharides. Particles
in which the carbohydrates are covalently appended to proteins are known as glycoproteins,
while those in which the starches are covalently connected to lipids are known as glycolipids. A
few sugars are edible by people and thusly give a vital source of energy, while others are inedible
and accordingly do not provide energy. The experiment seeks to analyze carbohydrates and
perform tests to obtain results that will be examined. Through observation, the group shall
perform the general test for carbohydrates, iodine test for polysaccharides, qualitative tests for
sugars and the corresponding sketch of mucic acid crystals based from the test solution
experimented. According to Wikibooks, the simplest carbohydrates are the monosaccharides,
also known as simple sugars. Disaccharides are double sugars, consisting of two
monosaccharides joined by a covalent bond. Carbohydrates also include polysaccharides, which
are polymers composed of many sugar building blocks. The name "carbohydrate" is derived
from 'hydrates of carbon', and they arise from photosynthesis, where they exist as products.

Analysis

The General Test for Carbohydrates is to be performed as the first experiment. The group
acquired 1% solutions of glucose, sucrose, arabinose, starch and cotton suspension then for each
1 mL of the sample a single drop of Molisch Reagent is to be poured in the 6 different samples in
six separate test tubes. After, cautiously pour mL of Hydrogen Sulfate down the tube’s side to
form a layer under the water. Then, observe the purple color at the junction of two liquids. Once
it is turned a green color it may be disregarded. Molisch's test is a complex chemical test for the
presence of carbohydrates, based on the dehydration of the carbohydrate by sulfuric acid or
hydrochloric acid to produce an aldehyde, which condenses with two molecules of phenol
(usually α-naphthol, though other phenols (e.g. resorcinol, thymol) also give colored products),
resulting in a red- or purple-colored compound. The group obtained results under molisch test as
the glucose having the mixture separated containing a dark color in the middle and a dark
brownish at the bottom with precipitate floating atop. For sucrose, floating particles are visible
on top, middle is a dark brown and the bottom is a clear dark brown. For arabinose, clearer light
brown at the bottom with brown in the middle and floating particles. The starch sample showed
an almost clear in the middle, almost black at the bottom and little number of particles suspended
on top. Lastly, the cotton suspension sample resulted to a very light green below while white part
is visible in the middle with brown floating particles. A deep violet coloration is supposed to be
produced at the junction of two layers but with the group’s obtained results it did not achieve the
purple color. If it had the shade of a deep purple this is due to the formation of an unstable
condensation product of beta-naphthol with furfural (produced by the dehydration of the
carbohydrate). Obtaining such results did not coincide with the expected result it must be due to
inaccuracy of the measurement of the chemicals added or there is no dehydration of carbohydrate
present at all.

The second experiment was performing the Anthrone Test. The Anthrone test is also
another general test for all carbohydrates. In this test also, carbohydrate gets dehydrated when
react with conc. H2SO4 to form furfural.with the procedure starting as the group placed three
drops of the sample in six separate small test tubes then quickly adding ½ mL Anthrone reagent
then mix carefully. Observe its color for a span of several minutes to half an hour. If milkiness
appears, dilute cautiously with 50% hydrogen sulfate. For the glucose sample, the group resulted
to a black liquid, the sucrose also having a dark colored liquid but is darker than glucose, the
arabinose a dark liquid that has a physical property of the color dark green almost black, the
starch a dark green color and lastly the cotton suspension sample which had a lime green
appearance. A positive test is obtained if the results showed a green color. Only the arabinose,
starch and cotton suspension had a positive result among the five samples.

The third test is the iodine test for polysaccharides is another test for carbohydrates that
Iodine staining distinguishes starch (a polysaccharide) from monosaccharides, disaccharides, and
other polysaccharides. The basis for this test is that starch is a coiled polymer of glucose. First,
add one drop of iodine solution to each of 2 drops of 1 % solutions of starch, dextrin, gum-arabic
and agar-agar since agar-agar is not available it is excluded in the experiment. After, observe the
changes. The starch resulted to a black middle part and yellowish surroundings inside the test
tube and the dextrin had a brownish orange or copper-like color and the gum-arabic responded to
a light yellow color. Iodine interacts with these coiled molecules and becomes bluish black.
Other non-coiled carbohydrates do not react with iodine. Therefore, a bluish black color is a
positive test for starch, and a yellow-ish brown color or no change in color is a negative test for
starch. The three samples of the group obtained negative results for starch since the color
responses to iodine shows otherwise.

The next experiment is the qualitative tests for sugars which had the test solutions of 5%
solutions of glucose, fructose, arabinose, maltose and sucrose; 5% solutions of galactose and
lactose all in all are seven samples. For benedict’s, Barfoed’s. Seliwanoff’s and Orcinol Tests the
procedure are all the same but is performed in different tests. The first process is to mix a single
drop of a test solution and a single drop of the reagent in 13 separate micro test tubes, heat in
boiling water bath and observe once there is visible results among any one of the test tubes
record and remove all from the water bath.

The Benedict’s test is a test to determine whether or not the carbohydrate contains a free
aldehyde or ketone group. In Benedict's reaction, copper is reduced and the product forms
a red precipitate. A Benedicts reagent contains sodium bicarbonate, sodium citrate and copper
sulfate. The first one that had a visible result is the fructose which turned orange meaning there
is a presence of reducing sugar. If the saccharide is a reducing sugar it will reduce Copper ions to
Cu oxide, a red precipitate.
The Barfoed’s test can differentiate monosaccharides from disaccharides and
polysaccharides since with the conditions of lower pH and shorter incubation time, only
monosaccharides can react fast enough to reduce copper ions. The group witnessed a change in
color first in lactose which turned to red with tiny precipitate indicating monosaccharides, no
color change or weak change will indicate disaccharides and polysaccharides.

The Seliwanoff’s test is used to differentiate between ketoses and aldoses. The reagent is
a solution of resorcinol in concentrated HC1. The acid when heated along with a sugar will
produce furfural or hydroxymethylfurfural, which further reacts to give a red color. Ketoses react
more rapidly than aldoses and thus the reaction time is a means of separation or detection. The
test solution that had a visible result was sucrose that had a cherry red color. Meaning, the
presence of ketoses is observed and sucrose gives a positive ketohexose test. When reacted with
Seliwanoff reagent, ketoses react within 1-2 minutes forming a cherry red condensation product.

The Orcinol’s test or also known as the Bial’s test is used to perceive the presence of
furanoses which are five-membered rings. These sugars, or compounds containing them, react
with Bial's reagent to give a green or olive colored solution. Furan rings contain five carbons but
sugars with furan rings can contain more carbons outside the ring, and all sugars with a furan
ring will react in Bial's test. The pentose furanoses will react with Bial's reagent to
form green solution, as the hexose furanose will react to form olive/brown solution. Bial's
contains orcinol (5-methyl- 1,3 dihydroxybenzene), the parent compound of the litmus dyes in
concentrated HCl. A muddy brown to gray product. The group detected the first visible change
in color in the test solution fructose which had the presence of hexoses. The furfurals formed
produces condensation products with specific colour.

Before all the experiments, Mucic Acid is performed in which concentrated HNO3 is
heated along with an aldose sugar to give a dicarboxylic acid. Nitric acid is able to oxidize the
terminal groups of aldoses, but leaves the secondary hydroxyl groups unchanged. The
dicarboxylic acid formed from galactose is called mucic acid and is insoluble in cold aqueous
solution. Those acids formed from the other common sugars are soluble in H20. Hence, the
formation of the insoluble precipitate is an indication of the presence of galactose.

The formation of a white precipitate indicates the presence of galactose in the


carbohydrates. From the results gathered, mucic acid only formed when the sugars galactose or
lactose were used as test samples. Glucose also forms crystals but it is soluble in an aqueous
solution, therefore it is not mucic acid. The aldehyde and primary alcohol groups are oxidized to
carboxyl groups in the reaction of glucose to HNO3 forming Saccharic acid.
Conclusion

Therefore, the tests performed aided in various identification tests for carbohydrates.
Also, the group acquired the skill to distinguish reducing and non-reducing sugars. For each test
and its corresponding result even though end product is a negative test or a result that is not
expected there is still a learning that the group acquired. The tests also aided us in the
identification for the reducing sugars found in the samples given. These results helped us in
understanding the ways disaccharides bond and how they are broken down to form the
monosaccharides.

Literature cited

For Website:

Luckie, Douglas. “Lab 1.” Vedas, 22 Aug. 2018, msu.edu/course/lbs/145/luckie/Lab1.html.

Open Books for an Open World, 22 Aug. 2018,


en.wikibooks.org/wiki/Structural_Biochemistry/Organic_Chemistry/Carbohydrates.

Structural Biochemistry/Organic Chemistry/Carbohydrates.” Music Theory/Modes - Wikibooks, 7.


ANALYSIS OF CARBOHYDRATES, people.umass.edu/~mcclemen/581Carbohydrates.html.

Hernandez, G. (n.d.). Mucic Acid Test. Retrieved from


http://www.academia.edu/22832264/Mucic_Acid_Test