BIO103 Lab
(1 credit hour)
SUMMER 2024
Dept. of Biochemistry & Microbiology
Experiment 5
Benedict test for determination of reducing sugars.
Benedict's test to Determine the Presence of Reducing Sugars
• Carbohydrates are the body’s most important and readily available source of energy. The
two major forms of carbohydrates are:
⮚ Simple sugars (simple carbohydrates), such as fructose, glucose and lactose, found in
nutritious whole fruits.
⮚ Starches (complex carbohydrates), found in foods such as starchy vegetables, grains,
rice, breads and cereals
• Carbohydrates are the main fuel source for some cells, especially, those in the brain,
nervous system and red blood cells.
• Muscles also rely on a dependable supply of carbohydrate to fuel intense physical activity.
Yielding on average 4 Kcal/gm, carbohydrates are a readily available fuel for all cells, both
in the form of blood glucose and that stored in the liver and muscles as glycogen.
Functions of carbohydrates
• Major functions for living systems
i. The principal function of carbohydrates, is to fuel cell activities with a ready source of energy.
• Through photosynthesis plants make carbohydrates (glucose). Those carbohydrates
form the foundation for almost all ecosystem’s energy source on Earth.
ii. Deoxyribose is a building block of DNA.
iii. Used in the synthesis of cell membrane & cell wall.
iv. Carbohydrates are also involved in cell-cell recognition.
v. Macromolecular carbohydrates function as food reserves (glycogen in animals).
Deoxyribose
Principle
• The purpose of this experiment is to investigate the presence of simple sugars in
various food products.
• Benedict's reagent is used for testing the presence of reducing sugars. This
includes all monosaccharides and certain disaccharides, e.g., mannose, lactose and
maltose.
• Sucrose is disaccharide present in sugarcane, however it is not a reducing sugar.
• A reducing agent donates electrons during a redox reaction and is itself oxidized.
The aldehyde functional group is the reducing agent in reducing sugars.
• Reducing sugars have either an aldehyde functional group or have a ketone group
in an open chain form, which can be converted into a carboxylic group.
Principle (cont.)
• In hot alkaline solutions, reducing sugars reduce the blue Copper (II) ions to brick
red Copper (I) oxide precipitate.
• As the reaction proceeds, the color of the reaction mixture changes progressively
from blue to green, yellow, orange and finally red.
• The coloration developed and the amount of precipitate formed depends upon the
amount of reducing sugars present. Hence, in most conditions, a sufficiently good
estimation of the concentration of glucose and equivalent reducing sugars present in
a sample can be obtained.
• Water plus Benedict's reagent is a negative control for the sugar test. It
demonstrates a negative test result (no sugar present). Carbohydrate sample plus
Benedict's reagent is a positive control for the sugar test.
Methodology
• Apparatus
Procedure
⮚ Test tubes
• Take 1ml of the apple juice provided in a clean test tube.
⮚ Water bath
• Add 2ml of Benedict’s Solution to each test tube.
⮚ Spatula
• Leave the test tubes in the hot water bath and note your
⮚ Dropper
observation.
⮚ Hot water bath
• A positive test with Benedict's reagent is shown by a color change
from clear blue to a brick-red precipitate.
• Reagents/Solvents • To prepare a negative control, repeat steps 2-3 using distilled water
⮚ Benedict’s reagent instead of sample solution (i.e., Apple juice).
⮚ Test sample (Apple juice)
⮚ Distilled water
Benedict’s Test
https://www.youtube.com/watch?v=k1DOadNdPY4
https://www.youtube.com/watch?v=HxevnEdU9AI
Thank you!
