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Definition
Uses
Limitations
Principle
 USES

DEFINITI LIMITATI
ON ONS

BARFOED’S TEST
 Barfoed’s test

DEFINITI USES LIMITATI

ON ONS
 This reaction can be  This test cannot be
Is a chemical test used used for disaccharides, used to detect sugar in
to detect the presence but the reaction would urine as urine contains
of monosaccharides be very slow. Cl– ions, which might
which detects reducing  This test is used to interfere with the
monosaccharides in identify reducing reaction.
monosaccharides and  If a higher
the presence of
distinguish the concentration of
disaccharides. reducing disaccharides disaccharides is
from reducing present in a sample, it
monosaccharides. might give a positive
NOTIFICATION

1
PRINCIPLE
PRINCIPLES
The Barfoed reagent is made up of
copper acetate in a dilute solution of
acetic acid. Since acidic pH is
unfavorable for reduction,
monosaccharides, which are strong
reducing agents, react in about 1-2 min.
However, the reducing disaccharides
take a longer time of about 7-8 minutes,
having first to get hydrolyzed in the
acidic solution and then react with the
reagent. Once the reaction takes place,
thin red precipitate forms at the bottom
of the sides of the tube. The difference
in the time of appearance of precipitate
thus helps distinguish reducing
PRINCIPLES
Barfoed’s Test is a copper reduction
test in which heating a mixture of a
monosaccharide in a given time
interval results in the conversion of
cupric ions in the Barfoed’s reagent
to cuprous ions with the formation of
copper (I) oxide seen as a brick red
precipitate in the mixture. Unlike
Benedict’s test, copper reduction in
Barfoed’s test occurs in an acidic
medium (pH 4.6) rather than an
alkaline medium.
PRINCIPLES
The Barfoed's reagent is
similar to Benedict's
reagent except that the pH is
lower (around 4.5), and heating
time is reduced to two minutes.
Benedict's test would determine
if the sample is a reducing
sugar, and Barfoed's test would
determine if it is a
monosaccharide or disaccharide.
 Contents 2
1. Objectives
2. Materials
3. Procedures
4. DATA AND RESULTS

5. Questions and answers

6. GENERALIZATION/SYNTHESIS
  To detect reducing carbohydrates.
 To distinguish reducing
OBJECTIVES
monosaccharides from
disaccharides.
 To distinguish between
monosaccharides and
disaccharides using the Barfoed’s
test
MATERIALS
MALTOSE
BARFOED’S
REAGENT
GLUCOSE
TEST TUBES

1 2 3 4
 Test tube stand
Water bath

PIPETTE
MEASURING
CYLINDER

5 6 7 8
PROCEDUR
1. Measure 3 mL of
freshly prepared
5. Observe the
Barfoed’s reagent
into labelled test
3. Place the test appearance of
tubes into a color in the test
tubes.
boiling water bath
tubes.
and heat for 3
minutes
2. Add 1 mL of the
test solution to 4. Remove the
Barfoed’s reagent. tubes from the
bath and allow
to cool.
 6. Noted own the 8. Do not heat the tubes
longer than 3 minutes, as
time taken for the a positive test can be
appearance of color obtained with
in the tubes. disaccharides if they are
heated long enough.

7. Formation of red or
yellow to orange
precipitate is a positive
test for reducing
monosaccharides.
 QUESTIONS
4. How to distinguish if the sample
1. How does the Barfoed’s contains reducing monosaccharides
or reducing disaccharides? What
test work? color indicates positive result?

5. Why do disaccharides
2. What makes Barfoed’s presence take longer time to
Test significant? form red precipitate than
monosaccharides?

3. Why did we use this in 6. What reaction takes

the barfoed’s test ? place in Barfoed’s Test?
ANSWERS
 Answers
1. Barfoed's test reaction is based on the reduction of
cupric acetate by reducing monosaccharides and
reducing disaccharides. Reduction of cupric acetate
produces cuprous oxide which gives a brick red
precipitate. Monosaccharides usually react in about 1-

ADD subtitle
3 min and produce a red precipitate.
2. This test is used to identify reducing monosaccharides
and distinguish the reducing disaccharides from
reducing monosaccharides.
3. The Barfoed reagent is made up of 0.33% molar
solution of neutral copper acetate in a dilute solution
of 1% acetic acid. Since acidic pH is unfavorable for
reduction, monosaccharides, which are strong
reducing agents, react in about 1-2 min. However, the
reducing disaccharides take a longer time of about 7-8
 ANSWERS
4. The difference in the time of appearance of precipitate
thus helps distinguish reducing monosaccharides from
reducing disaccharides. The presence of red precipitate
detects the presence of reducing monosaccharides in

ADD subtitle
the sample. If the color appears within the first few
minutes, the sample contains reducing
monosaccharides. However, if the color appears later
than the first 3 minutes, the
sample is of reducing disaccharides.
 Answers
5. The reaction with disaccharides is slower because
disaccharides have to get hydrolyzed first and then
react with
the reagent cupric acetate to produce cuprous oxide.
This

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delay in the development of the color of in case of
disaccharides is due to slow reduction of disaccharide
during
the reaction.

6.
Aldoses and ketoses can reduce cupric ions even in
acidic conditions. Reducing monosaccharides react with
Barfoed’s reagent much faster than disaccharides and
produce a copious amount of red precipitate of copper (I)
 Answers

ADD subtitle
Figure 1.
Interpretation of
Barfoed’s Test
Reaction
 CONCLUSION
Barfoed’s test is a biochemical test specific for
monosaccharide (reducing) sugars in solution. Due to the weakly
PENYEBAB
acidic nature of Barfoed's reagent, it is reduced only by
monosaccharides. Positive result for monosaccharides is the
DAMPAK
appearance of red precipitate within 1 to 2 minutes. If no
precipitate formed, it indicates the presence of PENANGGULANGA
disaccharide. Only
monosaccharide gives an immediate red precipitate in Barfoed’sN
test which is the glucose as we can observe in the experiment. On
the other hand, maltose did not show any changes in color as it is
classified as disaccharide.
Other monosaccharides are galactose and fructose.
 Barfoed’s test Precautions:

Precautions are very

necessary to be taken
before performing
this test. Without
these precautions,
there might make
mistakes and at the
results could be a
negative one.
 Barfoed’s test Precautions:

1
As compare to the
original sugar solution,
barfoed’s reagent should
be added in more
quantity. This ensures the
good reaction and a
positive result.
Barfoed’s test Precautions:
2
It is very necessary to
keep track of the time.
Make sure that there is a
digital stop watch
beside while performing
the test. Timing is
everything in this test
and you might lose the
experiment if not taken
care of the time. Give
boiling the exact time as
mentioned in the
procedure.
Barfoed’s test Precautions:

3
To check whether the
precipitates are
formed or not, remove
the test tube regularly
from the bath. Put the
test tube again and
again in the bath if
those are not formed.
  https://www.youtube.com/watch?v=yQf
MqvOxPrc
 https://www.onlinebiologynotes.com/barf
References:
oeds-test-objective-principle-reagents-pr
ocedure-and-result/