BARFOED’S TEST is a chemical test used to detect the presence of monosaccharides which detects

reducing monosaccharides in the presence of disaccharides. This reaction can be used for disaccharides,

but the reaction would be very slow. In addition, it is a biochemical test to detect monosaccharide

(reducing) sugars in solution, devised by the Swedish physician C. T. Barfoed (1815–99). Barfoed's

reagent, a mixture of ethanoic (acetic) acid and copper (II) acetate, is added to the test solution and

boiled. If any reducing sugars are present a red precipitate of copper (II) oxide is formed. The reaction

will be negative in the presence of disaccharide sugars as they are weaker reducing agents.

Principle of Barfoed’s test:

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 monosaccharides from reducing disaccharides.

Interpretation of Barfoed’s Test

 The presence of red precipitate detects the presence of reducing monosaccharides in 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.

Uses of Barfoed’s Test

 This test is used to identify reducing monosaccharides and distinguish the reducing

disaccharides from reducing monosaccharides.

Limitation of Barfoed’s Test

 This test cannot be used to detect sugar in urine as urine contains Cl– ions, which might

interfere with the reaction.

 If a higher concentration of disaccharides is present in a sample, it might give a positive result.

References and Sources
Tiwari A. (2015). Practical Biochemistry. LAP Lambert Academic Publishing.
3% – https://www.coursehero.com/file/70797005/Lab-Report-3-Carbohydratedocx/
2% – https://www.coursehero.com/file/p6hnhh1n/2-Barfoeds-Test-Barfoeds-test-is-used-to-distinguish-
between-reducing-mono-and/
2% – https://generalchemistrylab.blogspot.com/2011/12/barfoeds-test-for-monosaccharides.html
1% – https://www.coursehero.com/file/pml2bh/6-Place-the-test-tubes-in-a-boiling-water-bath-for-10-
minutes-to-develop-the/
1% – https://medicalstudyzone.com/barfoeds-test/
1% – https://allmedicalstuff.com/benedicts-test/