Barfoed’s Test

Vinuya, Cresence Joy M.

Calma, Ma. Katrina C.
Quiwa, Kimberly
Lintag, Ma. Cristina

This study aims to detect reducing sugar and to distinguish monosaccharides from reducing disaccharides. A Barfoed’s test
was used to distinguish reducing monosaccharides from disaccharides by controlling pH and time heating. A different
carbohydrates was used to test in the barfoed’s reagents such as; glucose, galactose, fructose, xylose, maltose, sucrose and
distilled water. The study shows that upon heating 2-5 minutes among the the different carbohydrates solutions glucose,
fructose, xylose and galactose has a positive reaction indicates the presence of a reducing monosaccharide and resulting to
the red color, while the maltose, sucrose, lactose and distilled water showed negative in this test and resulting the absence
of color which indicates that this set of carbohydrates are not monosaccharides. On the other hand, on prolonged heating
disaccharides can also give positive test. Therefore, we conclude that in testing the barfoed’s test time is very important in
distinguishing the monosaccharides and disaccharides.
Keywords:Barfoed’s Test,Barfoed’s Reagent,Monosaccharide,Disaccharide

Barfoed's Test is a qualitative test used to actually brick red solution with the copper acetic acid and
differentiate between a monosaccharide and acidic acid. They normally show up inside five (5)
disaccharide. A barfoed's reagent is a blue-green minutes. If they show up inside five minutes, it
color solution which is composed of copper sulfate implies that monosaccharide is present in the
(CuSO4) and acetic acid solution which provides solution.The red precipitate come from the
the acidic condition. Carbohydrates being tested reaction between the reduction of copper (II)
by barfoeds are monosaccharides and acetate to copper (I) oxide (Cu2O).A
disaccharides. Monosaccharides are monosaccharide is a better reducing sugar
carbohydrates composed of one sugar unit. All because they have aldehyde group but not all in
monosaccharides have the same general formula disaccharides. When a red copper (II) oxide
of (CH2O) which designates a central carbon precipitate formed, it indicates the presence of
molecule bonded to two hydrogen and oxygen. reducing sugar. The reaction will be negative in
An example of these are glucose (fig 1) and the presence of disaccharide sugars because they
fructose (fig 2). On the other hand, disaccharides are weaker reducing agents.This experiment aims
is a combination of two monosaccharide. An to distinguish monosaccharide from reducing
example of these are maltose (fig 3) and sucrose disaccharide.
(fig 4). In barfoed's test monosaccharide gives
 must be used),Distilled water for neutralization,
Glucose, Fructose, Maltose, Sucrose,
Figure 1 Xylose,Lactose, and Galactose.(Precaution:It
may cause burns or irritation to skin.Avoid
contact with all solutions and wear face
mask,lab gown and gloves while performing this
experiment).

Figure 2
Procedure
One ml each of the carbohydrate samples
(glucose,fructose,maltose,sucrose,xylose,galactos
e,lactose) and distilled water with barfoed’s
reagent was placed in a test tube. Two ml
Figure 3 barfoed's solution was added to each test tubes.
Each test tube with samples was boiled in a water
bath that changed in color and the time took for
the changed to take place was recorded.
1 ml of each sample in test tubes

Figure 4

(+2ml barfoed's reagent, caution:

skin irritant. wear medical gloves)

Methods
Boil in water bath

Materials
The materials/equipment used in the Barfoed’s
Test were the following water bath
(Precaution:A briskly boiling water bath should
be used for obtaining reliable results),dry test
tubes,test tube rack, dropper , beaker, and
pipette.The samples and chemical solutions
used in the experiment were the following:
Barfoed’s Reagent(freshly prepared reagent
Observe the color
Results and Discussion
Table 1. shows the appearance of 8 samples
before and after adding barfoed’s solution and
after boiling.
that the first set of Carbohydrates samples are
monosaccharides,while the second set are not
monosaccharide.

APPEA APPEA APPEA

RANCE RANCE RANCE
Samples Before After After Time
adding adding Boiling
Barfoe BArfoe
d’s d’s
1. Glucose Clear Blue brick 3
Green red at minute
the s 40
bottom sec.
2.Fructose Clear Blue brick 2
Green red at minute (photo 1)
the s 25
bottom sec.
3. Maltose Clear Blue Blue
Green Green 7m
4.Sucrose Clear Blue Blue
Green Green 7m
5.Xylose Clear Blue brick 2
Green red at minute
the s 25
bottom sec
6.Lactose Clear Blue Blue 7m
Green Green
7.Galactose Clear Blue brick 2
Green red at minute
the s 50
bottom sec
8.H20 Clear Blue Blue 7m (photo 2)
Green Green

Barfoed’s Test is copper reduction test in which

Glucose,Fructose,Xylose,and Galactose showed a heating a mixture of a monosaccharide in a given

changed in color from blue green to brick red time interval results in conversion of cupric ions in

(photo 1).The blue green color of Barfoed’s reagent to cuprous ions with formation

Maltose,Sucrose,Lactose,and Distilled water with of copper (I) oxide ,which is seen as a brick red

Barfoed’s solutionn remained unchanged after precipitate in the mixture. Barfoeds reagent is

heating(photo 2).The changed in color indicates composed of solution of copper sulphate and
acetic acid.The acetic acid provides the acidic
condition.When monosaccharides are heated with
barfoed’s reagent the color will changed into brick
red as because copper ions are being reduced to
copper oxide within 5 minutes.On the other hand ,
some disaccharides will take more than 5 minutes
or more to form red color precipitate of copper
oxide.Hence monosaccharide and (some)
disaccharide can be distinguished on the basis of
the time taken to form the red color precipitate.
In this test , fructose, galactose, xylose and
glucose which are monosaccharide showed a
positive result in barfoed’s test
1).Therefore, they formed a brick
color .Maltose and Lactose are
monosaccharide because they showed a negative
result within 5 minutes.For further information ,
these samples were boiled more than 5 minutes
and as the minutes pass ,these samples slowly
showed some changes. As a result both can give
a positive barfoed’s test.However the rate at
which monosaccharide and disaccharide react are
different.Monosaccharide react fast while the
disaccharide react slowly .Therefore, this forms
the basis of barfoed’s test.
References
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not gE-LjNyBXuyhda8
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U-O3mqPvVYGK__KVf5dPwN6OU-
Kbp7134f5AdRJs-25_0#google_vignette₩
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between monosaccharides and
disaccharides? What is a sugar unit?.
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https://www.quora.com/What-is-the-
difference-between-monosaccharides-and-
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unit?fbclid=IwAR3AIU4ZkqlAjR6MCOPIHSU
wylR-
5vgAmg3bhBpblzzVOh5rPp24RGVLC9wLC9

Journal:
Jahangirpuria, H. D.,Makwana S. A. , & C. G.
Patel (2017).Identification of carbohydrates.
The World Journal of Engineering and Applied
Science,(3),5-6.