Experiment (5):
Estimation of glucose by Benedict qualitatively
Benedict’s solution is a deep-blue alkaline solution used to test for the presence
of the aldehyde functional group, – CHO. Benedict’s solution can be used to test
for the presence of reducing sugars such as glucose in urine.
Benedict's qualitative reagent consists of:
CuSO4: provides the oxidizing Cu+2 ions.
Na2CO3: provides the alkaline medium necessary for the enolization of
carbohydrate.
Sodium citrate: prevents precipitation of Cu+2 ions as CuCO3 by forming
sodium cupric citrate complex which is slightly soluble complex that
dissociates slowly to give continuous supply of Cu+2 ions.
Principle of Benedict’s test
When reducing sugars are mixed with Benedict’s reagent and heated, a
reduction reaction causes the Benedicts reagent to change color. The color
varies from green to dark red (brick) or rusty-brown, depending on the amount
of and type of sugar. The copper (II) ions in the Benedict’s solution are reduced
to Copper (I) ions, which causes the color change.
The red copper (I) oxide formed is insoluble in water and is precipitated out of
solution. As the concentration of reducing sugar increases, the nearer the final
color is to brick red and the greater the precipitate formed.
Procedure of Benedict’s test
1. Approximately 1 ml of sample is placed into a clean test tube.
2. 2 ml (10 drops) of Benedict’s reagent is placed in the test tube.
3. The solution is then heated in a boiling water bath for 3-5 minutes.
4. Observe for color change in the solution of test tubes or precipitate
formation.
Result of Benedict’s test:
Positive Benedict’s test: Formation of a reddish precipitate within three
minutes and this indicate the presence of reducing sugars. Example: glucose.
- If the color upon boiling is changed into green, then there would be 0.1 to 0.5
percent sugar in solution (+).
- If it changes color to yellow, then 0.5 to 1 percent sugar is present (+++).
- If it changes to orange, then it means that 1 to 1.5 percent sugar is present (++++).
- If color changes to red, then 1.5 to 2.0 percent sugar is present.
- And if color changes to brick red, it means that more than 2 percent sugar is present
in solution (+++++)
Negative Benedict’s test: No color change (Remains Blue) and this indicate the
absence of Reducing sugars. Example: Sucrose.
