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Date: ……/..../….
PRACTICAL NO:- 1
PERFORM BENEDICT’S TEST
THEORY
Benedict's test is a chemical test that is used to test for the presence of reduced sugars within an
analytical test. Thus, simple carbohydrates that contain an aldehyde or free ketone functional
group are detected using this test. The test is basing itself upon Benedict's Reagent (also called
Benedict's solution) which is a complex mix of sodium carbonate, sodium citrate, and the
pentahydrate of copper(II) Sulfate.
If exposed to sugars that are reduced, the reactions will be triggered by Benedict’s reagent which
causes a brick-red-colored precipitate that is an indication of Benedict’s test is positive.
OBJECTIVE
To find the amount or presence of reducing sugar present in the mixture.
To determine the concentration of glucose in the solution. It is quantifiable.
PRINCIPLE
When Benedict’s solution and simple carbohydrates are heated, the solution changes to orange
red/ brick red. This reaction is caused by the reducing property of simple carbohydrates. 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. This
accounts for the precipitate formed. As the concentration of reducing sugar increases, the nearer
the final colour is to brick-red and the greater the precipitate formed. Sometimes a brick red
solid, copper oxide, precipitates out of the solution and collects at the bottom of the test tube.
Sodium carbonate provides the alkaline conditions which are required for the redox
reaction. Sodium citrate complexes with the copper (II) ions so that they do not deteriorate to
copper(I) ions during storage.
Complex carbohydrates such as starches DO NOT react positive with the Benedict’s test unless
they are broken down through heating or digestion (try chewing crackers and then doing the
test). Table sugar (disaccharide) is a non-reducing sugar and does also not react with the iodine
or with the Benedict Reagent. Sugar needs to be decomposed into its components glucose and
fructose then the glucose test would be positive but the starch test would still be negative.
REQUIREMENTS
Test solutions: 5 % Glucose, 5 % Sucrose
Benedict’s reagent: CuSO4.5H2O solution with Sodium carbonate and sodium citrate
Prepared By: - Ashok C. Keshari
HOD, Food & Dairy Tech.
NLPI, Bardibash
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Water bath
Dry test tubes
Pipettes
PROCEDURE
Preparation of Benedict’s Reagent
One liter of Benedict’s Reagent can be made using a mixture of 17.3 grams of copper
pentahydrate sulfate (CuSO4.5H2O), 100 grams sodium carbonate (Na2CO3) as well as 173
grams sodium citrate, in distillated water (required amount). This is because the copper(II)
sulfurate functions as a source for Cu2+ ions. The sodium carbonate acts as an alkaline medium
as well as the sodium citrate form complexes when it comes in contact with the Cu2+ ions.
Distilled water is utilized as solvent.
Pureness of Benedict’s reagent may be verified through heating in the test tube. The absence of
any change in the blue colour of the solution after heating indicates that the reagent is of pure.
Testing for Reducing Sugars
1. Approximately 1 ml of sample is placed into a clean test tube.
2. 2 milliliters (10 drops) of Benedict’s Reagent (CuSO4) is put into the testing tube.
3. After that, the solution will be heated by a water bath for about 3-5 minutes.
4. Check for changes in color inside the test tube or the formation of precipitate.
Result Interpretation of Benedict’s Test
If the color after boiling turns green, then there will there be 0.1 or 0.5 percent sugar in
the solution.
If the color changes to yellow Then 0.5 to 1 percent sugar levels are present.
Prepared By: - Ashok C. Keshari
HOD, Food & Dairy Tech.
NLPI, Bardibash
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If it turns orange, it indicates that between 1 and 1.5 percent sugar has been present.
If the color changes from red to black, 1.5 or 2.0 percent sugar levels are present.
If the color changes from brick red to red, it signifies there is more sugar than in the
solution.
Positive Benedict’s Test: Develops reddish precipitate within three minutes. Reducing
sugars present. Examples: Glucose
Negative Benedict’s Test: no change in color (Remains in blue). Sugars that reduce
sugars are not present. Example: Sucrose.
OBSERVATIONS
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RESULT AND DISCUSSION
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Prepared By: - Ashok C. Keshari
HOD, Food & Dairy Tech.
NLPI, Bardibash
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Precautions:
1. Handle the acids like concentrated sulfuric acid with care.
2. Always use droppers to take reagents from the reagent bottles.
3. While heating the reaction mixture do it carefully.
Limitation of Benedict’sTest
The weaknesses of Benedict’s test can be seen as follows:
False-positive reactions during the test may be observed when certain drugs are such as
streptomycin, isoniazid and salicylates penicillin, P-aminosalicylic acid are obtained.
The chemical compounds present in concentrated urine can decrease Benedict’s reaction,
which comprises creatinine, urate, and ascorbic acid (the reduction is only slight).
Uses of Benedict’s
It can be utilized in the search for unknown carbohydrate in biochemistry analysis.
It is possible to detect diabetes mellitus by a clinical diagnosis.
Because the procedure for preparing the benedict reagent is easy and straightforward,
executing the test is inexpensive and easy to conduct.
The amount of time needed is extremely minimal.
The test may be both quantitative and qualitative.
Prepared By: - Ashok C. Keshari
HOD, Food & Dairy Tech.
NLPI, Bardibash
