SECONDARY EVIDENCE FOR TEST FOR CARBOHYDRATES

BENEDICT'S TEST

The procedure for Benedict’s test is:

1. Put approximately 1 mL of Benedict's reagent into a clean, dry test tube.

2. Add 2-3 drops of the liquid sample into the test tube.
3. Heat the content of the test tube for approximately 3 minutes in a heated water bath.
4. Formation of a red precipitate of cuprous oxide indicates the presence of a reducing
sugar. Low amounts of reducing sugars appear as a green or orange precipitate

BARFOED’S TEST

1. Take 1 ml of a given sample in a clean, dry test tube. The concentration of disaccharides
sample (if used) should not exceed 1% (w/v).
2. Take control of 1 ml of distilled water in another tube.
3. Add about 2-3 drops of Barfoed’s reagent to both the tubes and mix them in a vortex.
4. Keep the test tubes in the water bath for 1-2 minutes. The boiling should not be done for
more than 2 minutes as the disaccharides might hydrolyze into monosaccharides and
give a positive result.
5. Observe the appearance of colour in the test tubes.
6. Note down the time taken for the appearance of colour in the tubes.

EVALUATION
Benedict's test and Barfoed's test are both chemical assays employed to detect the presence of reducing
sugars. Benedict's test is a general test, utilising Benedict's reagent with copper ions, which form a
coloured precipitate when reduced by sugars. This test is broadly applicable to all reducing sugars, without
specific discrimination between mono- or disaccharides. In contrast, Barfoed's test is designed to
distinguish between monosaccharides and disaccharides. It employs Barfoed's reagent, a
copper-containing solution like Benedict's reagent, but it is more selective, producing a quicker reaction
with monosaccharides compared to disaccharides. The specificity of Barfoed's test makes it a useful tool
for differentiating between simple and double sugars.
SECONDARY EVIDENCE FOR TEST FOR CARBOHYDRATES

theory.labster.com. (n.d.). Benedict’s Test: Procedure - Labster Theory. [online] Available at:
https://theory.labster.com/benedicts_procedure/.

Sapkota, A. (2022). Barfoed’s Test- Definition, Principle, Procedure, Result, Uses. [online]
Microbe Notes. Available at: https://microbenotes.com/barfoeds-test/.