SPECIFIC TEST FOR CARBOHYDRATES

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Barfoed’s Test
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What is Barfoed’s Test?

 It is a chemical test for the detection of MONOSACCHARIDE

 It is based on the reduction of copper II acetate into copper I

oxide which forms a BRICK-RED PRECIPITATE

 DISACCHARIDE may also react but the reaction is slower

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Why is the heating procedure limited to
3 minutes?

 On prolonged heating disaccharides can also give a positive

result.

 That is why it is specify to heat the sugar solution for not more
than 3 minutes in order to be sure that only monosaccharide will
react because they react faster than disaccharide
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How does Barfoed’s Reagent work?

 Barfoed’s Reagent is composed of Acetic acid and copper (II)

acetate.

 The ACETIC ACID provides the acidic condition of the reagent

for the reaction to occur

 MONOSACCHARIDE will reduce the copper (II) acetate in the

barfoed’s reagent forming a brick-red precipitate copper (I) oxide

 This test involves Redox Reaction

Reducing monosaccharides are oxidized by the copper ion
in the solution to form carboxylic acid and brick-red
precipitate of copper (I) oxide within 3 minutes. Reducing
disaccharides undergo the same reaction but in much slower
rate.
GLUCOSE POSITVE
Glucose gives a positive reaction because it is a polyhydroxy aldehyde group
that normally forms cyclic hemiacetal that has the property to reduce copper ion
to copper (I) oxide.
FRUCTOSE POSITIVE
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 Fructose give off a positive result although it is a ketose sugar

because it is an alpha-hydroxy ketone
LACTOSE NEGATIVE
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 LACTOSE IS ALSO A REDUCING SUGAR BUT IT IS A

DISACCHARIDE WHICH MEANS IT REACTS SLOWER THAN
MONOSACCHARIDE THAT IS WHY IT IS SPECIFY IN THE
PROCEDURE NOT TO OVER EXTEND THE HEATING
PROCESS TO MORE THAN 3 MINUTES.
SUCROSE NEGATIVE
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 SUCROSE IS NOT A REDUCING SUGAR BECAUSE IT

CANNOT REVERT TO THE OPEN-CHAIN FORM THAT
WOULD PROVIDE THE ALDEHYDE GROUP NEEDED TO
REDUCE CUPRIC IONS AND IT CONTAINS ACETAL LINK
INSTEAD OF HEMIACETAL THEREFOR IT IS LOCKED IN ITS
CYCLIC FORM.
 Barfoed’s Test Benedict’s Test Fehling’s Test

Barfoed’s test uses copper Benedict’s test uses copper Fehling’s Test uses Rochelle
acetate and acetic acid sulfate, sodium carbonate and salt and copper sulfate
sodium citrate

Uses acetic acid to make an Uses sodium carbonate to Uses sodium carbonate to
acidic environment for the make an alkaline environment make an alkaline environment
reaction to occur for the reaction to occur for the reaction to occur

Used to distinguished Used to distinguished aldose Test the presence of aldehydes

monosaccharide vs. vs. ketose monosaccharide but not ketones
disaccharide except for fructose

It is only brick-red colored The precipitate may vary in Only red precipitate is obtained
precipitate which is obtained color depending on the
concentration of sugar it may
be green, yellow, orange or red
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What test is more accurate?

 If you want to test specific sugar solution if it is monosaccharide

or a disaccharide the most accurate test is Barfoed’s Test.

 If you want to identify the concentration of sugar in the solution

the most accurate test is the Benedict’s Test.

 If you want to test the presence of aldehyde in the sugar solution

then the most accurate test is the Fehling’s Test
 SPECIFIC TEST FOR CARBOHYDRATES
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SELIWANOFF’S
TEST
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What is Seliwanoff’s Test and How does
it works?

 It is a chemical test which distinguishes between ketose and

aldose sugars.

 It relies on the principle that when heated ketoses are more

rapidly dehydrated than aldoses

 Which then reacts to the resorcinol that yield a cherry-red hue

complex color as the positive result
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Composition of the Seliwanoff’s Reagent

 The seliwanoff’s reagent is composed of Hydrochloric

acid, and Resorcinol

 HCL is used to dehydrate keto sugars

 The dehydrated keto sugar then reacts to resorcinol

to create a cherry-red hue complex color
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Sucrose and Fructose

 Sucrose obtain a positive result because sucrose or

table sugar is composed of fructose and glucose
wherein fructose is a keto sugar that can be
dehydrated easily.

 Therefor the sucrose can still yield cherry-red hue

complex because of its composition.

 Same goes with fructose

Fructose will undergo dehydration and will become hydroxymethyl furfural
and will react to the resorcinol that is present in the reagent that will give
off cherry-red hue coloration
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Glucose

 Did not yield a positive result because it is an aldose

sugar. Aldehyde group do not react with resorcinol
that is present in the reagent.
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Comparison between Seliwanoff’s Test
and Molisch Test

 Seliwanoff’s test is a chemical test which only reacts to ketone

group while Molisch test on the other hand reacts with aldehyde
group.

 But both of the test relies in the principle of dehydration with the
use of strong acid (in the case of seliwanoff’s test is the
resorcinol in the molisch test the sulfuric acid)