BIOCHEMISTRY LABORATORY WORKSHEET

Name: _____________________________________ Rating: ________________

Course, Year and Section: ______________________ Instructor: _____________
Group Number: ______________________________ Date: ________________

General Reactions of Carbohydrates

EXPERIMENT 3

Objectives:
At the end of the experiment, students should be able to:
1. Describe the properties and structures of carbohydrates
2. Interpret the visual evidence of the different tests for carbohydrates

Reagents/Materials:
1% fructose Molisch reagent iodine solution
1% glucose Benedict’s reagent Concentrated H2SO4
1% glycogen Fehling’s A reagent 1% Ba(OH)2
1% starch Fehling’s B reagent
1% xylose 1% sucrose

Procedures:
I. General Reactions of Carbohydrates
A. Molisch Test
1. Prepare 4 clean test tubes, label each and put 1 mL of the following into each test
tube: 1% glucose, 1% xylose, 1% sucrose, and 1% starch.
2. To each test tube add 2 drops of Molisch reagent and mix thoroughly.
3. Incline the test tube and carefully add 1 mL of concentrated H2SO4 to flow down the
side of the test tube. Do not shake. Observe the color formed at the junction of two
layers and record your data.

B. Reducing Property
B.1. Fehling’s Test
1. Prepare Fehling’s reagent by mixing 2 mL of Fehling’s A, 2 mL of Fehling’s B, and 8
mL of distilled water in a big test tube.
2. Prepare 4 clean test tubes.
3. Place 1 mL of Fehling’s reagent in a test tube and 5 drops of 1% glucose solution.
Heat in a boiling water bath. Observe and record the color of the precipitate.
4. Repeat step 3 using the other test solutions (1% xylose, 1% sucrose, and 1%
starch).

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 BIOCHEMISTRY LABORATORY WORKSHEET

B.2. Nylander’s Test

1. Mix 15 drops of 1% glucose with an equal amount of Nylander’s solution in a test
tube. Heat for 5 minutes in a boiling water bath. Note the color produced
2. Repeat using the other test solutions (1% xylose, 1% sucrose, and 1% starch).

B.3. Benedict’s Test

1. To 1ml of Benedict’s reagent in a test tube, add 1 ml of 1% glucose solution. Heat
in a boiling water bath for 3 minutes and allow to stand. Observe and record your
data. Note the time required to show a green or red precipitate.
2. Repeat step 3 using the other test solutions (1% xylose, 1% sucrose, and 1%
starch).

B.4. Moore’s Test

1. In a test tube, add 2 mL of 6N NaOH to 1 mL of 1% glucose. Heat in a boiling water
bath. Note changes in color and odor.
2. Repeat the test using 1% sucrose.
3. In another test tube, add 2 mL of 1% Ba(OH)2 to 1 mL of 1% glucose. Heat in a boiling
water bath. Note changes in color and odor.
4. Repeat the test using 1% sucrose.

Note: No part of this Biochemistry Laboratory worksheet can be reproduced, distributed, shared, or posted/uploaded on the Internet or in any other means without
the permission of the authors. Any violation will be dealt with as academic dishonesty and will be meted with a sanction.
 BIOCHEMISTRY LABORATORY WORKSHEET

Name: _____________________________________ Rating: ________________

Course, Year and Section: ______________________ Instructor: _____________
Group Number: ______________________________ Date: ________________

Reactions of Carbohydrates
EXPERIMENT 2

supposedly sucrose should turn positive but due to its non reducing
I. General Reactions of Carbohydrates nature it resulted negative and the starch is supposed to be negative
because it does not have free aldehyde and ketone groups but it
A. Reaction with Acids - Molisch Test potentially occurred that it's positive due to presence of carbohydrates

Test solution Reagents Result Interpretation

monosaccharide and a reducing sugar,
1% glucose nt Positive (purple ring) contains a free aldehyde group that reacts
age with the Molisch reagent

h Re monosaccharide and a reducing sugar,

1% xylose
olisc Positive (purple ring) possesses a free aldehyde group that reacts
with the Molisch reagent
sM non-reducing disaccharide composed of
1% sucrose rop glucose and fructose units, lacks free

2d O4
Negative (no purple ring) reducing ends does not react with the

2S
Molisch reagent

1% starch L H polysaccharide composed of glucose units,

can still give a positive result if it contains
1m Positive (purple ring) free reducing ends that allows starch to react
with the Molisch reagent

B. Reducing Property
B.1. Fehling’s Test

Test solution Reagents Result Interpretation

contain free aldehyde or ketone groups that
1% glucose B
Positive (reducing sugar, forms a
red precipitate) react with Fehling's reagent

and
A r contain free aldehyde or ketone groups that
1% xylose g’s ate
Positive (reducing sugar, forms a
red precipitate) react with Fehling's reagent
in w
hl d
1% sucrose Fe stille Negative (non-reducing disaccharide, no does not have free reducing ends and
therefore does not react with Fehling's
i reaction)
Ld
reagent

8m does not have free reducing ends and

1% starch Negative (non-reducing polysaccharide,
no reaction) therefore does not react with Fehling's
reagent

B2. Nylander’s Test

Test solution Reagents Result Interpretation

1% glucose Black precipitate monosaccharide and a reducing sugar

t
1% xylose gen Brown precipitate monosaccharide and a reducing sugar
ea
1% sucrose r'sR No precipitate (remains clear) non-reducing disaccharide, do not have free
de aldehyde or ketone groups

ylan
1% starch N No precipitate (remains clear) non-reducing polysaccharide, do not have
free aldehyde or ketone groups

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the permission of the authors. Any violation will be dealt with as academic dishonesty and will be meted with a sanction.
 BIOCHEMISTRY LABORATORY WORKSHEET

sucrose supposedly give a negative result since it is a non-reducing

B3. Benedict’s Test disaccharide, but then it can undergo hydrolysis in the presence of heat
and acid to break down into glucose and fructose, which are reducing
sugars that makes it positive.

Test solution Reagents Result Interpretation

1% glucose nt
orange-colored precipitate reducing sugar

age
1% xylose Re
ct's orange-colored precipitate reducing sugar

di
1% sucrose ene non-reducing disaccharid hydrolysis in heat

LB darker orange-colored precipitate and acid, breaking down into constituent

monosaccharides (glucose and fructose)
1 m which are reducing sugars
no change in color or formation of a
1% starch precipitate non-reducing polysaccharide, does not have
free aldehyde or ketone groups

B4. Moore’s Test

Test solution
Reagents Result Interpretation
The yellow color observed may be due to the breakdown of glucose under alkaline
glucose A 2 mL 6N NaOH Maillard reaction, which can occur between
reducing sugars like glucose and the alkaline
conditions, which can produce various
volatile compounds, some of which may
environment provided by NaOH. resemble the smell of spoiled milk.
The orange color observed may be due to the
spoiled milk odor could be a result of the
glucose B 1% Ba(OH)2 formation of colored complexes between
glucose and Ba(OH)2. Maillard reaction occurring between glucose
and the alkaline Ba(OH)2 solution
no significant color change was mentioned, it No specific odor was mentioned, indicating
sucrose A 6N NaOH suggests that the reaction between sucrose
and NaOH did not result in a noticeable color
that there might not have been any
significant odor-producing reactions
change in this particular experiment. between sucrose and NaOH.
no significant color change was mentioned, it No specific odor was mentioned, indicating
sucrose B 1% Ba(OH)2 suggests that the reaction between sucrose
and Ba(OH)2 did not result in a noticeable
that there might not have been any
significant odor-producing reactions between
color change in this particular experiment. sucrose and Ba(OH)2.

Note: No part of this Biochemistry Laboratory worksheet can be reproduced, distributed, shared, or posted/uploaded on the Internet or in any other means without
the permission of the authors. Any violation will be dealt with as academic dishonesty and will be meted with a sanction.