Name: Jovan Paul G.

Delda CHEM 104A Lab Schedule: 12:00 – 4:00 PM M-F Course &
Year: BSN 1 – A Name of Lab Instructor: Jemimah Joy Guarin

CARBOHYDRATES
Activity No. 3

I. PURPOSE: To be able to know some general properties of carbohydrates.

II. APPARATUS: Bunsen burner, test tubes, beaker (600 ml), graduated cylinder (25 ml),
dropper, vials, test tube holder

III. MATERIALS: Pinch of arabinose, fructose, glucose, sucrose, and starch 5 ml

Fehling’s A, 5 ml Fehling’s B
Drops of: 0.01 M arabinose, 0.05% starch, 15% alpha-naphthol in
alcohol, concentrated H2SO4, Seliwanoff’s reagent, Phloroglucin
reagent, iodine solution, 7% starch solution

IV. PROCEDURE:

A. Solubility

Test the solubility of each arabinose, fructose, glucose, sucrose, and starch in water.
Add only a pinch of the solute to be tested in 2 ml of the solvent.

B. Reactions of Carbohydrates

1. Molisch Test

Prepare four tubes containing 1 ml of water, 1 ml 0.01 M glucose, 1 ml 0.01 M

sucrose, and 1 ml 0.05% starch respectively. To each of the four solutions add 2 drops of
a 15% solution of alpha-naphthol in alcohol. Mix well. Slowly pour down the side of each
tube 3 ml of concentrated sulfuric acid. Note the colors of the rings obtained after some
time. The first tube serves as a control. In some cases, as in polysaccharides, it may
require a longer time to develop to test.

 A 15 percent solution of alpha-naphthol alcohol and 3 ml of

concentrated sulfuric acid are added to a test tube labeled M1, and
the color of the solution changes to red-orange.
 The color of the glucose is yellow-orange and has clear color in the
lower portion of the test tube after adding 2 drops of a 15 percent
solution of alpha-naphthol alcohol, mixing well, and adding 3 ml of
concentrated sulfuric acid
 The color of M3 or sucrose becomes yellow-orange.
 M4 or the starch becomes reddish-orange

2. Seliwanoff’s Test
  The test reagent dehydrates ketohexoses to form 5-hydroxymethylfurfural. 5-
hydroxymethylfurfural further reacts with resorcinol present in the test
reagent to produce a red product within two minutes.

Place in each of the four test tubes, 5 ml of Seliwanoff’s reagent. To these tests,
tubes add respectively 1 ml of water, 1 ml of 0.01 M glucose, 1 ml of 0.01 M fructose, and
1 ml of 0.01 M arabinose. Immerse all test tubes in boiling water at the same time.
Observe the odor changes occurring in each tube during a 5-
minute period of heating.

3. Tollen’s Phloroglucin Test

Measure off 5 ml portions of phloroglucin solution into each of four test tubes. To
each tube add respectively 1 ml of water, 1 ml of 0.01 M glucose, 1 ml of 0.01 M fructose,
and 1 ml of 0.01 M arabinose. Immersed all test tubes in boiling water at the same time.
Observe the color changes for two minutes during the first 15 minutes of heating then
observe at the end of 30 minutes. This reaction may be used to differentiate pentoses
from hexoses.

 T1 - it changes color to red and orange.

 During the heating process, T2 fiercely boils and turns crimson.
 T3 - The color changes to dark violet or red-violet.
 T4 - its color changes to a red-maroon color.

4. Fehling’s Test

Into 5 separate test tubes place 1 ml of each of the following solutions: 0.01 M
glucose, 0.01 M fructose, 0.01 M sucrose, 0.7% starch solution, and water. To all 5 test
tubes add 1 ml of Fehling’s solution A and Fehling’s solution B. Shake and immerse all
tubes in boiling water and heat for 15 minutes. Note the resulting colors and the odor in
which the reaction occurs.

5. Fermentation Test

Into four different test tubes place 5 ml of the following solutions: 0.01 M glucose,
0.01 M galactose, 0.01 M sucrose, and 0.01 M starch. To each test, the tube adds an
equal amount of 20% suspension of ordinary Baker’s yeast. Let it stand for 30 minutes
and observe any bubbling formation.

 Fr1 – It changes to a yellow-orange tint and develops bubbles.

 Fr2 – Its upper and middle portions are white, while the lower test
tube is clear and forms bubbles.
 Fr3 – It turns pale yellow and the top has a few bubbles.
 Fr4 – The test tube changes to white and has several bubbles on top.
 6. Iodine Test

To 5 ml of 0.7%, the starch solution adds a few drops of dilute iodine solution until
a good color develops. Results? Warm the tube very gently. Results? Cool the test tube
and observe.

 Add a few drops of starch to one of the spot plate's wells. Iodine reagent
should be added in two drops. A vivid blue-black complex should result
from the reaction of starch and iodine.

QUESTIONS:

1. What can be said about the solubility of carbohydrates in water?

 Simple carbohydrates are small polar molecules, containing several - OH
functional groups, which makes them hydrophilic and they dissolve well in
water. Polysaccharides, also called complex carbohydrates, are large
nonpolar molecules, and they are not hydrophilic.
2. Account for the differences in results obtained during the fermentation of glucose,
sucrose, galactose, and starch.

 They all have bubbles formed. The glucose color turned into yellow-orange,
and the color of the sucrose turns to white yellow and has a clear color in
the lower part of the test tube. The galactose color in the test tube is lightly
yellow. The starch in the test tube has many bubbles formed and its color is
white.

3. Account for the results obtained for the iodine test for starch.

 Upon the addition of iodine solution in the starch its color turned into dark
blue or black purple and it has a brown-orange stain in the test tube.
4. Give the composition of the following reagents:
a. Molisch
 gm of α-naphthol in 25 ml of Ethanol 99% 

b. Seliwanoff’s

 The acid hydrolysis of polysaccharide and oligosaccharide ketoses yields simpler

sugars followed by furfural.
 The dehydrated ketose then reacts with two equivalents of resorcinol in a series
of condensation reactions to produce a molecule with a deep cherry red color.
 Aldoses may react slightly to produce a faint pink color.
Fructose and sucrose are two common sugars that give a positive test. Sucrose gives a
positive test as it is a disaccharide consisting of fructose and glucose.
Generally, 6M HCl is used to run this test. Ketose gets dehydrated faster and hence they
give the test faster. Aldoses react very slowly and give faint colors.

c. Phloroglucin

 Phloroglucinol is an organic compound with the formula C6H3(OH)3. It is a

colorless solid. It is used in
the synthesis of pharmaceuticals and explosives. Phloroglucinol is one of
three isomeric benzenetriols. The other two isomers
are hydroxyquinol (1,2,4-benzenetriol) and pyrogallol (1,2,3-benzenetriol).
Phloroglucinol, and its benzenetriol isomers, are still defined as "phenols"
according to the IUPAC official nomenclature rules of chemical compounds.
Many such monophenolics are often termed "polyphenols" by the cosmetic
and para-pharmaceutical industries, which does not match the scientifically
accepted definition.

d. Fehling’s A

 A is a blue-colored aqueous solution of copper (II) sulfate (CuSO4).

e. Fehling’s B

 A colorless aqueous solution of potassium sodium tartrate

(KNaC4H4O6·4H2O, also known as Rochelle salt) in an alkaline base like
sodium hydroxide (NaOH).

5. Give the principle of the following tests:

a. Molisch

 The dehydration of sulfuric acid into furfural is the fundamental component

of the Molisch test. One hydroxyl group is removed from the sugar
molecule when a sample containing carbohydrate molecules is treated with
strong hydrochloric acid and sulfuric acid. Water is released in place of the
hydroxyl group.

b. Phloroglucin test

 The furfural and phloroglucinol interaction, which results in a colorful

product with high molar absorptivity, is the basis for this test. For the
detection of lignin, a mixture of hydrochloric acid and phloroglucinol is also
utilized (Wiesner test).

c. Seliwanoff’s test

 Aldose and ketose sugars can be distinguished using Seliwanoff's test, a

chemical test. Sugar is a ketose if it has a ketone group in it. Sugar is an
aldose if it has an aldehyde group. This experiment is based on the idea
that when heated, ketoses dehydrate more quickly than aldoses.
d. Fehling’s test

 The reduction of the deep blue copper (II) solution to a red precipitate of
insoluble copper oxide allows for the detection of aldehydes but not
ketones in this test. Although known to be insensitive to aldehydes, the test
is frequently employed for reducing sugars. Fructose and acetoin, for
instance, both pass Fehling's solution test.