Worksheet CHEM1163L

Biochemistry Laboratory

Experiment 2
Properties of Carbohydrates

Names: Iona De Los Santos and Kyla Fernandez_____________ Schedule ________________

Write your correct observations and inferences/conclusions for each of the following:

A1. Molisch Test

Reagent’s Alpha-naphthol(C10H8OH) dissolved in ethanol(C2H5OH)
Composition
Observations: After combining sucrose, distilled water and Molisch as well as adding
sulfuric acid. After letting it sit for a few seconds a darker color began
forming at the bottom and eventually formed a ring in the solution.
Inferences: Molisch Test is used to test carbohydrates in a solution and to show it has
carbohydrates there will be a purple ring, and after testing sucrose it
shows a purple ring proving that there are carbohydrates.

A2. Seliwanoff’s Test

Reagent’s This reagent consists of resorcinol and hydrochloric acid
Composition
Chemical Test Sample Chemical Structure
Structure: Glucose C6H1206

Fructose C6H12O6

Arabinose C5H10O5

Observations: Test Sample Observations

Water The color of the water remained the same after it was
added with the reagent and heated in a water bath.
Glucose The color of this carbohydrate remained the same after
it was added with reagent and heated in a water bath.
Fructose The color of this carbohydrate changed into a cherry
red-complex after it was added with the reagent and
heated in a water bath.
Arabinose The color of this carbohydrate remained the same as
well after it was added with the reagent and heated in a
water bath.
Inferences: What is the principle behind this test?
The principle behind this test is that it used to distinguish a carbohydrate
with aldoses from ketoses. On treatment with concentrated acid, ketoses
dried out more rapidly to yield furfural derivatives and on condensation
with resorcinol, a cherry, red-colored complex forms.

What kind of sugars will give a positive result to this test?

Worksheet CHEM1163L
Biochemistry Laboratory

The presence of ketoses in a carbohydrate gives a positive result to this

test it is because the Seliwanoff reagent will give of a cherry, red-colored
complex only to those carbohydrates that contains a ketone group. Since
fructose has a ketone group in its structure therefore, the sample give a
positive result.

A3. Bial’s Orcinol Test

Reagent’s Mixture if Ferric Acid(H2FeO4), Hydrochloric acid(HCl), and

Composition orcinol(C7H8O2)
Observations: Test Sample Observations
Water It remains a yellow color showing that it is not a
pentose.
Glucose It showed a yellow color after a few seconds meaning
that it is a hexose sugar.
Fructose There was a negative reaction where in the solution
showed a yellow color meaning that it is a hexose sugar.
Arabinose It shows a positive reaction where in the color of the
solution turned a bluish tint showing that it is a pentose.
Inferences: What is the principle behind this test?
-the principle of this test is to simply be able to check for the presence of
pentose.

What kind of sugars will give a positive result to this test?

-the sugars that give a positive result to this test is pentose sugars, and
sugars like ribose and xylose.

A4. Fehling’s Test

Reagent’s Fehling’s compositions A=aqueous copper sulphate (CuSO4) and Fehling’s

Composition Solution B= alkaline sodium potassium tartrate(Rochelle Salt)
Observations: Test Sample Observations
Water It does not have any reducing sugars since it shows a
blue tint.
Glucose The glucose after a while shows a orange like color
meaning that it has reducing sugars.
Fructose It shows a somewhat dirty brown showing that it has a
reducing sugar in it.
Sucrose It shows a yellow substance lining at the bottom while
the solution is a green like color.
Starch Starch even after for a while it still shows a blue-green
tint.
Inferences: What is the principle behind this test?
-Is to look for reducing sugars.
Worksheet CHEM1163L
Biochemistry Laboratory

What kind of sugars will give a positive result to this test?

-The kinds of sugars that will give a positive result are reducing sugars,
examples are maltose and lactose.

A5. Benedicts Test

Reagent’s This reagent has a complex mixture of sodium carbonate, sodium citrate,
Composition and copper(II) sulfate pentahydrate.
Observations: Test Sample Observations
0.1M glucose The original color of the sample changed into a
brick red color
0.04 M glucose The color of the sample is blue with a little shade of
brick red
0.002M glucose The color did not change at all.
Inferences: What is the principle behind this test?
-When the Benedict’s solution and glucose are heated, the solution
changes its color into orange red/ brick red. This reaction happens
because of the reducing property of glucose. The copper (II) ions in the
Benedict’s solution are reduced to Copper (I) ions, which changes its
color.

Explain the difference in the amount of precipitate seen in the results.

-As the concentration of reducing sugar increases, the closer the final
color, brick-red, would reveal and the greater the precipitate formed. In
the sample containing 0.002 M glucose, it is still far from turning into a
brick red color because it has a low concentration of reducing sugar. But
as the amount increases such as 0.04 M to 0.1 M of glucose the brick red
color shade is starting to show until it becomes a solid color.

Provide a practical application of Benedicts Test

-A practical application of Benedict’s Test is when the solution can be
used to detect the presence of sugars in the urine. Further tests have to be
undergone once a reducing sugar is detected in order to find out which
sugar is present. Glucose is usually found in the urine but in very low
levels or absent. Glycosuria, a sugar that has an abnormally high amount
of sugar, can be also detected and are usually the result of high blood
sugar levels of untreated diabetes. Having these kinds of sugars can be an
indication of diabetes mellitus.

A6. Fermentation Test

Reagent’s This reagent consists of 20% suspension of ordinary Baker’s yeast and
Composition phosphate buffer( pH6.6 to 6.8 )
Observations: Test Sample Observations
Glucose There was a formation of bubbles and the height of
the mixture rose rapidly from its starting point. This
sample was the fastest one to rise among the four
samples.
Worksheet CHEM1163L
Biochemistry Laboratory

Galactose Small bubbles were seen and the height of mixture

rose slowly from its starting point. As a result, there
was a slight change of height from its initial to its
final point.
Sucrose Bubbles were noticed as well and the height of the
mixture rose quickly, catching up glucose. After
sometime, this sample and the glucose sample
almost have the same height of its final position.
Starch The presence of bubbles was there and the mixture
rose at the same rate as the galactose sample as
well as the final position of the height.
Inferences: What is the principle behind this test? What is the enzyme present in the
Baker’s yeast.
-The principle behind this test is that the action of organism on a
carbohydrate substrate causes the medium become acidic, which can be
detected with a pH indicator. A maltase, an enzyme that is present in
baker’s yeast breaks maltose into glucose and is responsible for
producing many glucose for yeast fermentation.

Explain the difference of the amount of bubbles produced by the different

test samples.
-The glucose and the sucrose samples were able to produce large amount
of bubbles because the enzymes in the yeast fermented the sugar
successfully causes to produce a lot of carbon dioxide. The more carbon
dioxide is produced, the more bubbles are formed. The galactose and the
starch samples were only able to produce small of amount of bubbles
since the enzymes in the yeast are unable to ferment them therefore, little
to no carbon dioxide was able to produce.

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