NAME: DATE:

GROUP NO:

PHINMA UNIVERSITY OF PANGASINAN

COLLEGE OF ALLIED HEALTH SCIENCES
DEPARTMENT OF NURSING

ACTIVITY NO. 10
TEST FOR CARBOHYDRATES

I. DISCUSSION
Classification and Structure of Carbohydrates
Simple Sugars (Aldoses and Ketoses, Pentoses and Hexoses)
Reducing and Non-Reducing Sugars
Name of Test, purpose of test, composition of reagent result

II. PROCEDURE

A. GENERAL COLOR TESTS

1. Molisch Test
To a test tube containing 2 ml. of carbohydrate solution (e.g. glucose, sucrose and starch)
add 1-2 drops of alpha-naphthol solution and mix. Incline the test tube and slowly and
carefully add about 1 ml. of concentrated sulfuric acid (CAUTION!) down the side of the
tube to form a layer below the sugar solution. Note the color formed at the junction of the
two liquids.

Results: A violet ring color was formed at the junction of two liquids.

2. Anthrone Test
To 5 drops of sugar solution in a test tube, Add 1 ml. each anthrone reagent. Mix
thoroughly by swirling. Records observe the color formed.

Results: A bluish green color complex was formed

B. TESTS FOR REDUCING SUGARS
1. Benedict’s Test
Add 2 ml. of Benedict’s reagent to 2 ml. of glucose, sucrose and starch
solutions in separate test tubes. Shake each tube thoroughly and place each tube in a
boiling water bath. Heat for 5 minutes and observe what happens. Record your results
in a table.
Substance Result (Color) Obtained
Tested
glucose red
sucrose blue
starch blue

2. Nylander’s Test
B. Nylander's test is a chemical test used for detecting the presence of reducing
sugars. Glucose or fructose reduces bismuth oxynitrate to bismuth under alkaline
conditions. When Nylander's reagent, which consists of bismuth nitrate, potassium
sodium tartrate and potassium hydroxide, is added to a solution with reducing sugars,
a ________(color) precipitate of metallic bismuth is formed. Fill in the blank the color
that is formed.

3. Barfoed’s Test
Add 1 ml. of barfoed’s reagent to 1 ml. of glucose, sucrose and lactose
solutions placed in separate test tubes. Heat in boiling water, take out after 10 mins
and see the record of the result in the table.

Substance Result (Precipitate /

None Precipitate - Color)
Obtained Tested
 glucose precipitate-red
sucrose non precipitate blue
lactose non precipitate blue

C. MUCIC ACID TEST FOR GALACTOSE AND LACTOSE

Mucic acid test is a test that is highly specific and is used for the detection of the
presence of galactose and lactose. It is also termed galactaric acid that is named after the
product of the reaction.

Objectives of Mucic acid test

To detect the presence of galactose and lactose in a given sample.
To distinguish between the galactose containing saccharides and other sugars.

PROCEDURE:
1. To 1 ml. of galactose and lactose solution in separate evaporating dishes,
add 5 ml. of conc. nitric acid. Evaporate on a steam bath under the hood to
a volume of 2 ml.

2. Allow to cool, cover the evaporating dishes with watch glasses and set
aside in your locker until the next laboratory period.

3. Add 5 ml. of distilled water to each dish, mix well with a stirring rod, and
pour the contents of the dishes into clean test tubes.

4. Examine the crystals under the microscope.

QUESTIONS:
1. What does the formation of crystals of the tube indicate?

The formation of crystals at the bottom of the tube indicates a positive result,
indicating the presence of galactose or its derivatives
2. What does the absence of such crystals indicate?

The absence of such crystals indicates a negative result, indicating that

 the sample doesn’t contain galactose or its derivative.
Other carbohydrates in the solution may still be present

D. SELIWANOFF’S TEST FOR KETOSES

Place 1 ml. each of solutions of maltose, fructose and glucose in separate test tubes.
Add 5 ml. of Seliwanoff’s reagent to each tube, mix and heat for exactly 30 sec in the boiling
bath, observe and record results at this time.

Results:

Maltose resulted negatively because no cherry red color was generated. Therefore, ketosis
is absent. Glucose resulted negatively as well because no cherry red color was formed. As a
result, ketosis is absent. Fructose, on the other hand, was associated positively since it
generated the hue cherry red; hence, ketosis is pressent.

E. TESTS FOR PENTOSES

A. Aniline Acetate Test

Place 3 ml. of arabinose in a test tube. Add 2 ml. of conc. HCI and carefully boil
the resulting solution for about 1 minute. Discontinue heating and hold a small piece of
filter paper moistened with aniline acetate over the mouth of the tube. Do not allow the
paper to rest on the mouth of the tube. Hold the paper, otherwise false results may be obta
Results:
With aniline acetate, it generates a vivid red hue on a paper placed over the mouth of the
reaction flask. A sheet of paper previously impregnated with aniline acetate is exposed to
the vapor of the sample solution. Furfural and aniline combine to produce a brilliant pink
hue. This signifies that the test resulted in a positive result, showing that the pentoses are
present.

B. Bial’s Test
To 1 ml. of Bial’s reagent in a test tube, add 1 ml. of arabinose and heat in a
boiling water bath for two mins. Observe and record the changes.
Results:
This test is only for pentoses. Hexoses usually react to green, red, or brown items. If the
Bial's Test is positive, it will become blue. For pentoses, all other hues imply a bad outcome.
REFERENCES:

http://dept.harpercollege.edu/chemistry/chm/100/dgodambe/thedisk/carbo/molisch/molisch.htm

https://handwiki.org/wiki/Chemistry:Nylander%27s_test
https://microbenotes.com
https://enacademic.com/dic.nsf/enwiki/3718875?fbclid=IwAR381crJ4QahECGptlhzMQob-
td1vmhnzvh60H3IE2SxO4fJQHJHd0SFDc4
https://byjus.com/questions/what-is-bials-test/

EXPERIMENT VIDEO/DISCUSSION

https://www.youtube.com/watch?v=0aWSHlK7Xtk&t=843s
https://www.youtube.com/watch?v=zHcmxBi_wG0
https://www.youtube.com/watch?v=Wq5uh0bVJzM&feature=youtu.be
https://www.youtube.com/watch?v=Ewe7i1D9lSQ&t=433s