Lab#: 2

Title: Testing for Reducing and Non-Reducing Sugars

Date: 13 February, 2022

Objective: to discover how reducing sugars can be detected using Benedict’s solution, how

this can be made semi-quantitative and how the test can be extended to register the

presence of non-reducing sugars as well.

Materials:

1. Large beakers to act as a water bath (500 cm 3 or larger)

2. Six test tubes

3. Test tube rack

4. Spatula

5. Two syringes or graduated pipettes for measuring 2 cm 3

6. Marker pen

7. 20 cm 3 Benedict’s solution

8. Distilled water

9. Solid glucose

10. Few cm 3 1% glucose solution

11. Solid sucrose

12. 0.1 mol dm -3 HCL

13. Solid NaHCO 3

14. Test tube holder

 15. Rubber bands

Procedures:

Testing for reducing sugars

1. A water bath (large beaker) was heated to about 75 C. This was set up before beginning

to prepare for testing.

2. A small spatula-load of glucose (which is a reducing sugar) was dissolved in

approximately 2 cm3 water and a roughly equal volume of benedict’s solution was added.

The solution was mixed thoroughly; then the test tube was put in the water bath and

maintained there for three to four minutes. More consistent results were obtained when

the test tube was gently agitated during heating.

3. The sequence of colour changes was observed and noted. The final colour sequence was

brick-red.

Making the test semi-quantitative

1. A serial dilution was set up with a 1% glucose solution. Six test tubes were labelled, 1-6,

with a marker pen, and 2 cm3 1% glucose was placed in the first test tube and 2 cm3

distilled water in the other five. 2 cm3 1% glucose was added to tube 2 and mixed

thoroughly before 2 cm3 was transferred to the next tube, until you reach tube 6. The

contents of tube 6 were mixed and then 2 cm3 was discarded. 2 cm3 benedict’s solution

was added and again mixed thoroughly.

2. All the tubes were placed in a boiling water bath and leave them for a standard time. (at

least three minutes)

 3. The tubes were removed and arranged in order, the colour against the concentration was

recorded.

Test for non-reducing sugars

1. A small spatula-load of sucrose was dissolved in half a test tube of distilled water, and

the solution was split equally between two test tubes labelled A and B. To A

approximately 1 cm 3 0.1 moldm-3 hydrochloric acid was added, but to B the same volume

of distilled water was added. Both tubes were heated in the boiling water bath for one

minute.

2. Both tubes were removed and cooled under a tap to approximately room temperature. To

tube A a small amount of sodium hydrogen carbonate powder was added. (Safety note:

the effervescence that follows can be vigorous if the tube is not thoroughly cooled or if

too much hydrogen carbonate is added). Small quantities of the powder were

continuously added until the addition of more does not result in effervescence. This

indicated that the acid has been neutralized and the alkaline conditions necessary for the

benedict’s test have prevailed.

3. An equal amount of benedict’s solution was added to each test tube and was heated for

standard time in the water bath. The test tubes were removed, and the final colours were

obtained.

Results:

Testing for reducing sugars

Light Blue → Green → Yellow → Orange → Brick red

Making the test semi-quantitative

 Test Tube 1 Test Tube 2 Test Tube 3 Test Tube 4 Test Tube 5 Test Tube 6

Initial colour Light Blue Light Blue Light Blue Light Blue Light Blue Light Blue

Final colour Brick Red Rusty Red Brown Moss Green Blue-Green Light Blue

Test for non-reducing sugars

A B

Initial Colour Clear Golden Yellow

Final Colour Clear Clear

Discussion:

Carbohydrates (also called carbs) are a type of macronutrient found in certain foods and drinks.

Sugars, starches and fibre are carbohydrates. The term saccharide refers to the unit structure of

carbohydrates. Carbohydrates are simple organic compounds that are aldehydes or ketones with

many hydroxyl groups added usually on each carbon atom not part of the aldehyde

or ketone functional group. The general chemical formula of carbohydrates is Cn (H2O) n. Not all

carbohydrates follow this formula and are slightly different in structure from this rule. There are

also compounds that seem to follow this rule but are not carbohydrates (e.g. formaldehyde).

They are an essential structural component of living cells. They are an important source of

energy for animals. Monosaccharides are the most fundamental type is the sugars. They

are glucose, galactose, and fructose. These simple sugars can combine with each other to form

more complex types. The combination of two simple sugars is called a disaccharide whereas

those consisting of two to ten simple sugars are called oligosaccharides, and those with a larger
number are called polysaccharides. Benedict's reagent (often called Benedict's qualitative

solution or Benedict's solution) is a chemical reagent and complex mixture of sodium carbonate,

sodium citrate, and copper(II) sulfate pentahydrate. It is often used in place of Fehling's solution

to detect the presence of reducing sugars. The more red a solution the more glucose (reducing

sugar) the solution contains

Source of Error:

The pipette for the CuSO4 touched one of the solutions and was therefore contaminated. The test

tubes touched the bottom of the water bath.

Conclusion:

It was discovered how reducing sugars can be detected using Benedict’s solution, how

this can be made semi-quantitative and how the test can be extended to register the

presence of non-reducing sugars as well.

Benedict´s Test for Reducing Sugars ( A&I)

Discuss reducing/non-reducing sugars (3 marks)

-include chemical reaction (redox) between reducing sugar ad Benedict´s solution [1]

-explain why non-reducing sugars do not react with Benedict´s [1]

-describe how Benedict´s test can be used semi-quantitatively [1]

Analysis of Results (7 marks)

-accurately calculate concentrations (%) of 6 glucose solutions [1]

-describe what is meant by condensation & hydrolysis with respect to sugars and starch [1]

-discuss the effect of hydrolysis of sucrose on the Benedict´s test [2]

-discuss the effect of hydrolysis of starch on the Benedict´s test [2]

-compare semi-quantitatively the amount of reducing sugar released after hydrolysis of both sucrose

and starch [1]

Accurate Sources of error/limitations/assumptions

-any two [2]

-less than two [1]