Benedict’s test is a simple chemistry test that is used to detect reducing sugars. Reducing sugars are carbohydrates that have a free aldehyde or ketone functional group in their molecular structure. These include monosaccharides such as glucose and fructose and disaccharides such as lactose and maltose. Benedict’s reagent, also known as benedict’s solution is used in Benedict’s test for detecting simple sugars such as glucose. It is a bright blue solution that is prepared by mixing copper sulfate pentahydrate, sodium carbonate, and sodium citrate in distilled water. In this article, we will learn about the benedict’s test in detail that includes the benedict test principle, benedict’s test procedure, benedict test reaction, and the benedict’s solution formula.
What is Benedict’s Test?
Benedict’s test is a chemical test that is used to check for the presence of reducing sugars in an analyte. Hence, simple carbohydrates that contain a free ketone or aldehyde functional group can be identified using this test. The benedict’s test for reducing sugars is based on the benedict’s reagent, which is also known as Benedict’s solution and is a complex mixture of sodium citrate, pentahydrate of copper(II) sulfate, and sodium carbonate.
When it is exposed to the reducing sugars, the reactions undergone through Benedict’s reagent form a brick-red precipitate that indicates a positive reducing sugar test. Take a look at the image of the benedict’s test colours of Benedict’s reagent that change from clear blue to brick-red and which are triggered by the exposure to reducing sugars.
Benedict’s test can also be used for checking the presence of glucose in a sample of urine. Since this benedict test for urine detects the presence of any aldehydes and α-hydroxy ketones and given that glucose is an aldose that has an open chain that forms an aldehyde group, the test gives a positive result if glucose is present in the analyte. However, a positive reaction is also given in the presence of homogentisic acid, ascorbic acid and several other reducing substances. Hence, a positive Benedict’s test does not always mean that the test subject is having diabetes.
Principle of Benedict Test
Benedict’s test is performed when the reducing sugar is heated with Benedict‘s reagent. The alkaline sodium carbonate present converts the sugar into a strong reducing agent called enediols. During the occurrence of the reduction reaction, the mixture changes its colour from blue to brick-red precipitate because of the formation of cuprous oxide Cu2o. Copper which is in its cupric Cu2+ or copper (I) form is then reduced to cuprous Cu+ or copper (II) form. Since the red-coloured cuprous oxide is insoluble in water, it is separated. If the concentration of the sugar is high, the colour of the resultant solution becomes more reddish, and, in turn, the volume of the precipitate increases.
Benedict’s Test Procedure
A mixture that contains the reducing sugar (about 8 drops of urine) and Benedict’s solution (approximately 5 ml) is heated in a test tube for around two minutes and is then allowed to cool. The colour of the mixture changes accordingly and results in precipitates. This indicates positive results. The results of the benedict’s reagent test are summarized in the table below.
Benedict’s Reagent Test Result
Observation of Color |
The Concentration of Reducing Sugar in g % |
Interpretation |
Blue, no change in colour |
0 |
No presence of reducing sugar |
Green Precipitate |
0.5 – 1 |
Presence of traceable amount of reducing sugar |
Yellow precipitate |
1 – 1.5 |
Presence of a small amount of reducing sugar |
Orange-red precipitate |
1.5 – 2 |
Presence of a moderate amount of reducing sugar |
Brick-red precipitate |
> 2 |
Presence of a large amount of reducing sugar |
Benedict’s Test Results
The test results for benedict’s test for glucose is shown below.
Limitation of Benedict’s Test
The limitations of Benedict’s test are as follows:
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False-positive reactions in the test can also be obtained if there are certain drugs present for example, salicylates, isoniazid, streptomycin, penicillin, and p-aminosalicylic acid.
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The chemicals present in the concentrated urine may reduce Benedict’s reaction which includes urate, creatinine, and ascorbic acid (the reduction is slight).
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