Seliwanoff’s

Test
GROUP 2
 Introduction
Seliwanoff’s test is a chemical test which
distinguishes between aldose and ketose sugars. If the sugar
contains a ketone group, it is a ketose. If a sugar contains an
aldehyde group, it is an aldose. This test relies on the principle
that, when heated, ketoses are more rapidly dehydrated than
aldoses. It is named after Theodor Seliwanoff, the chemist that
devised the test. When added to a solution containing ketoses,
a red color is formed rapidly indicating a positive test. When
added to a solution containing aldoses, a slower forming light
pink is observed instead.
 Introduction

The picture above shows the reaction of the Seliwanoff's Test.

 Objectives

• To detect the presence of ketohexoses

in a given sample

• To distinguish ketoses from aldoses

 Materials
Reagent
• Seliwanoff’s reagent: add 0.05% resorcinol (m-
hydroxybenzene) in 3 N HCl. Dissolve 50 mg
resorcinol in 33 ml concentrated HCl and make it
100 ml with water.
• Test sample
• Distilled water

Materials required
• Test tubes
• Test tube stand
• Pipettes

Equipment
• Water bath
 Procedure

1 2 3 4
Add 2 ml of Observe the
Take two clean, Keep both the test
Seliwanoffs’ formation of
dry test tubes and tubes in a water
reagent to both the color and note it
add 1 ml of the bath for 1 min.
test tubes. down.
test sample in one
test tube and 1 ml
of distilled water
in another as
blank.
 Data & Results

The formation of the cherry red-colored complex indicates a

positive result which means that the given sample contains
ketoses.

The absence of such color or the appearance of the color after

a prolonged period of time indicates a negative result which
means that the test sample doesn’t have ketoses.
 Data & Results
Test Result Color Present or Absent

Positive Cherry Red Ketoses present

Negative Clear/ transparent Ketoses absent

Positive Seliwanoff’s Test: If the color changes to red, then

your result is positive and keto sugar (Fructose and Sucrose) is
present inside the solution.

Negative Seliwanoff’s Test: If no red color appears or if a

faint pink color appears, you’re your result is negative and
Aldose sugar (Glucose) is present in the solution.
Questions & Answers
What are the uses of the
1 Seliwanoff’s test?

Seliwanoff’s color reaction is used in the

method for the colorimetric determination of fructose
in fermentation media. A modified version of this test
can be used for the determination of the concentration
of ketoses in a given sample.
Questions & Answers
What are the limitations of
2 Seliwanoff’s test?

The high concentration of glucose or other sugar

may interfere by producing similar colored compounds with
Seliwanoff’s reagent. Prolonged boiling can transform
glucose to fructose by the catalytic action of acid and form
cherry red-complex giving a false-positive result. This test is
a generalized test and doesn’t distinguish between specific
ketoses, and a separate test is required for the particular
ketose sugar identification.
 Questions & Answers
What are the principles of
3 Seliwanoff’s test?
The reagent of this test consists of resorcinol and concentrated HCl.
The acid hydrolysis of polysaccharides and oligosaccharides yields simpler
sugars. Ketoses are more rapidly dehydrated than aldoses. Ketoses undergo
dehydration in the presence of concentrated acid to yield 5-hydroxymethyl
furfural. The dehydrated ketose reacts with two equivalents of resorcinol in a
series of condensation reactions to produce a complex (not a precipitate), termed
xanthenoid, with deep cherry red color. Aldoses may react slightly to produce a
faint pink to cherry red color if the test is prolonged. The product and reaction
time of the oxidation reaction helps to distinguish between carbohydrates. Other
carbohydrates like sucrose and inulin also give a positive result for this test as
these are hydrolyzed by acid to give fructose.
 Questions & Answers
What are the reagents of the test?
4
The reagents consist of resorcinol and concentrated hydrochloric
acid:

• 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.
Questions & Answers
What are the sugars that give
5 positive results?

Fructose and sucrose are two common sugars

which give a positive test. Sucrose gives a positive test as it
is a disaccharide consisting of fructose and glucose.
Questions & Answers
What is the role of dilute HCl in
6 Seliwanoff's test?

The Seliwanoff Reaction, a well-known color

reaction for ketoses, is based in the fact that ketoses are
dehydrated more rapidly than aldoses to give a furfural
derivative. Further condensation with resorcinol in dilute
hydrochloric acid gives the color product.
 Conclusion / Synthesis
We therefore conclude that after the test being performed, we
observed that when a ketose is reacted with this reagent, it becomes
dehydrated and a cherry-red complex forms. Aldoses also react with this
reagent, but much more slowly than ketoses. This experiment is usually
experimented with fructose and sucrose but there are many sugars which give
a positive test. Once 5-hydroxylmethylfurfural is produced, it reacts with
resorcinol to give a dark red condensation product. Sucrose hydrolyzes to give
fructose, which eventually reacts to produce a dark red color. When
Seliwanoff’s reagent is reacted with a disaccharide or a polysaccharide, the
acid in the solution will first hydrolyze them into monosaccharides, and the
resulting monosaccharides can then be dehydrated. Disaccharides and
polysaccharides will therefore react slowly with Seliwanoff’s reagent.
 THANK YOU!
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