FACULTY OF HEALTH SCIENCES

ENV 416 PRACTICAL

NAME : .......................................................... DATE : .......................................

EXPERIMENT 4

TITLE : Identification of Carbohydrates

OBJECTIVES:
1. Set up control samples of known simple sugars and complex carbohydrates, run a series of three
tests on them; Benedict’s, Barfoed’s, and Iodine
2. Test several foods using the same tests, then by comparing to the results from the control samples
determine the presence, and type of carbohydrate in the food samples

Background Information:

An important group of organic molecules found in the foods we eat, are carbohydrates. Our bodies
break down carbohydrates as a source of energy. These carbohydrates come in two forms. The simplest
type being the simple sugars, monosaccharides and disaccharides. The monosaccharides, like glucose
and fructose, contain one sugar unit that can be in one of two forms: ring or open chain. The
disaccharides consist of two monosaccharides bonded together. Examples of common disaccharides are;
sucrose, maltose and lactose. Sucrose, the table sugar we buy from the store, is made up of the
monosaccharides glucose and fructose. Lactose is the sugar found in milk. Almost everyone has heard of
lactose intolerance, caused by a deficiency of the enzyme lactase which is responsible for breaking
lactose down into two monosaccharides. Breaking the disaccharides down into monosaccharides is
accomplished by enzymes in our bodies, but the bonds can also be broken by heat, or by a reaction with
acid.

The complex carbohydrates make up the second type of carbohydrates. The complex carbohydrates are
polymers of the monosaccharides in ring form. (A polymer is a molecule made up of many of the same
units bonded together to form a very large molecule.) Starch and cellulose are two types of complex
carbohydrates. Starch, in its two forms, amylose and amylopectin, can be broken down into
monosaccharides by the 60 digestive system, heat and acids. Cellulose on the other hand, though a
polymer of glucose, is bonded together in a slightly different manor and cannot be broken down in
the human digestive system. In some animals bacteria in the digestive tract breakdown the cellulose,
making it available to the animal for use.

Today we will set up an experiment to test known carbohydrate samples (the controls) and samples
from several foods (the unknowns). The tests we will use are:

I. Benedict’s test: this test detects the presence of aldehyde groups using the Cu2+ in a basic
solution. It will react with both monosaccharides and disaccharides containing aldehyde groups
with equal speed. From the structure above you can see that glucose has an aldehyde group
when it is in its open chain form. Appearance of an orange color from the presence of Cu2O
indicates a positive result. See the reaction equation below:
1
II. Barfoed’s test: this test also uses copper, but in an acetic acid solution. While Benedict’s reagent
reacts with both monosaccharides and disaccharides at the same speed, Barfoed’s reagen
reacts more quickly with the monosaccharides than the disaccharides. Because of this
difference, this test can be used to tell if your sample contains monosaccharides or
disaccharides. For this test you need to look for the colored precipitate and record how long it
took to develop.

III. The third test we will use is a simple iodine test. When iodine is added to a solution and a dark
blue color is produced, starch is present. The iodine fits itself into the helix structure of the
amylose in such a way that the position of the molecule produces a dark blue color.

Procedure:
1. Measure out 5ml of each of the samples, controls and food samples, into labeled
test tubes. (remember to observe any differences between solutions and record your
observations in your notebook.)

2. Add 8 drops of Benedict’s reagent to each tube, mix gently, and place tubes in a boiling water
bath for 3 minutes. After you remove and cool the tubes record the results, colors observed, in
your notebook.

3. Measure out 2ml of each of the samples, controls and food samples, into labeled test tubes.
Add 2ml of Barfoed’s reagent to each, mix gently, and place in a boiling water bath. Make sure
you record the time tubes were placed in bath. Then watch for color change and remove tubes
as the color change occurs. Remember to record the time of color change as well as the color
change!! Remove all tubes after 30 minutes, and examine sides of tubes for presence of
precipitate after carefully pouring liquid from tubes.

4. While your tubes for step 2 are in the boiling water bath one partner can be setting
up and running the Iodine test. Place 2ml of each of the samples, controls and food samples,
into labeled test tubes. Then add a couple of drops of the iodine solution to each, mix gently
and record any color change in your notebook.

By comparing the test results of the controls to the test results of the food samples determine if
carbohydrates are present in the samples, and if they are what type, monosaccharide,
disaccharide, starch. Remember only positive tests give you information that you can use.