CARBOHYDRATE LAB

Bring one Food item to be tested for Carbs on Lab day to receive
some Extra Credit!!!!Best are fruits, vegetables, cereals, dairy,
honey, juices, chips, candies
I. BACKGROUND ON CARBOHYDRATES
Carbohydrates are organic molecules that contain the elements ________________,
_________________ and _____________________ exclusively. They belong to the general class called
polyalcohols because they have multiple ________________ group next either one ________________ or
______________on Carbons 1 or 2. Carbohydrates comprise about 60% of our daily caloric intake and are
the major source of metabolic energy. Carbohydrates are categorized as monosaccharides, disaccharides, or
polysaccharides.

Monosaccharides: Are the simplest carbohydrates and serve as the building blocks for _____ and
___________ saccharides. Give three examples of monosaccharides:
_____________________________________________________________________________All
monosaccharides are reducing sugars, which means they ________________________ other molecules
while being __________________________ at the same time. Even the ketose Fructose is considered a
reducing sugar even though you learned that a ketogroup cannot be further oxidized. This is how: Fructose in
the presence of an oxidizing agent relocates its _____________ group from carbon #2 to carbon #1,
essentially turning into glucose and then undergoes oxidation.

Disaccharides:

In a disaccharide, _______ monosaccharides are linked together via a

_____________________bond. Lactose, also commonly known as _________sugar, is found in milk and milk
products. Lactose is made of a ___________________ and _________________________. People who
lack the enzyme ________________________ are known to be lactose _________________. Sucrose,
common table sugar, is composed of ____________________________ and _________________________.
Unlike the monosaccharides, not all disaccharides are reducing sugars. The only disaccharide
that is not reducing is ___________________

Polysaccharides: Polysaccharides are polymers composed of ___________________ monosaccharide

units. The most common polysaccharides have glucose as a building block, and are different only in their
________________________.. Starch composed of the polysaccharides ____________________ and
_________________________, is the major form of glucose storage in plants. Amylose (~ 20% of starch) is
a linear chain of glucose while amylopectin (~ 80%) is a branched polymer of glucose. Glycogen is the major
form of glucose storage in animals, and is found in the liver and muscle. Glycogen, like amylopectin in plants,
is a branched polymer of glucose. In mammals the breakdown (hydrolysis) of glycogen helps maintain
_________________________glucose levels between meals. Cellulose is an unbranched chain of glucose
and is the major ____________________________________ of plants. In Cellulose the glucose molecules
are connected via a _________________________ glycosidic bond. Humans do not possess cellulose and
therefore cannot digest and utilize the calories stored in cellulose. Such undigestable carbohydrates provide
the major source of ___________________ in our diet.

Common Tests for characterizing sugars

Benedicts test for reducing sugars: Qualitative Benedicts reagent (Cu2+) reacts
with reducing sugars to form a red precipitate (Cu+). Quantitative Benedicts produces
a sliding color scale ranging from green to gold to red depending on the amount
(concentration) of the reducing sugar present in the sample.
Draw in the redox brackets and label them
Label all compounds
Reduced copper has the color: ______
Oxidized copper has the color: ______

Seliwanoffs test for ketoses: Seliwanoffs test can be used to distinguish 6 carbon
monosaccharides (hexoses) that have a ketone group from those hexoses that contain
an aldehyde group. With ketoses the reagent produces a deep red color rapidly, while
with aldoses a light pink color develops over a longer period of time.
Iodine test for polysaccharides: Iodine interacts with the structure of amylose to
produce a deep blue-black complex. The polysaccharides amylopectin, glycogen, and
cellulose, interact with iodine to give red to brown colors. Glycogen gives a reddishpurple color with iodine. Mono and di- saccharides do not interact with iodine to form
dark colored complexes.
Hydrolyzing Di- and Polysaccharides
Disaccharides and polysaccharides can be hydrolyzed (broken down) into their
constituent monosaccharides. In the laboratory hydrolysis can be achieved by reacting
the di- and polysaccharides with acid. Acid hydrolysis of disaccharides will produce the
constituent monosaccharides. In our bodies the hydrolysis of polysaccharides (except
cellulose) is carried out by two amylase enzymes that are found in our saliva or secreted
by the pancreas.

II.

PRE-LABORATORY QUESTIONS

Fill in the blanks in the background portion of this lab.

Read through the procedure part of the lab, so you understand which
procedure is used to identify, or find absence or presence of a sugar when
its dissolved as an aqueous solution.
1. Which method can be used to distinguish between a glucose and fructose
solution?
2.

Which method can be used to distinguish between sucrose and lactose

solution?

3. Which method can be used to distinguish between a glucose and amylose

solution?
4. You made three solutions containing either glucose, fructose and sucrose but
forgot to label them. Show in which order you would carry out tests to identify
without doubt which solution contains which sugar.

5. How can di- and polysaccharides be broken down into their constituent
monosaccharides?
6. Draw the hydrolysis Rx of sucrose, structures and names.

7. Based on your knowledge about the different tests for Carbohydrates, predict the
outcome (final color) of the Benedicts test and the Seliwanoffs test for
Sucrose, and Glucose

8. Write the formula (not to be mistaken with structure!) for glucose, fructose,
sucrose, maltose and lactose. What did you find out?

III.

PROCEDURE

Start your hot water bath!!!

Materials:
One large beaker (250-400 mL), plastic well plate, micro-centrifuge tubes and clips,
boiling water bath, floating test tube rack, hot plate, beral pipettes, and plastic 24-well plate,
various sugar solutions, Benedicts, Seligwanoffs and Iodine Test solutions.
A. BENEDICTS TEST FOR REDUCING SUGARS
Place 5-6 drops of glucose, fructose, sucrose, lactose, starch, unknown and
H2Oin micro-centrifuge tubes.
Add 9 drops of Benedicts reagent to each sample, then cap and gently shake
the closed tube to mix.
Heat all the tubes in a boiling water bath for 3-4 minutes.
The formation of a greenish to reddish-orange color indicates the presence of a
reducing sugar. If the solution stays blue then there was a non-reducing sugar
the solution
Record in table in Data Report Sheet
B. SELIGWANOFFS TEST FOR KETOSES
Place 2-3 drops of glucose, fructose, sucrose, lactose, starch, H2O and unknown
in new micro-centrifuge tubes.
Add 10 drops of Seliwanoffs reagent to each sample, then cap and gently shake
the closed tube to mix. The reagent contains concentrated HCl. Use
carefully.
Place the test tubes in a boiling hot water bath and note the time. After 5
minutes, observe the colors in the tube. The formation of a deep red color
indicates the presence of a ketose. Record your results as a deep red color
change, pale red/pink change or no change.
Record in data record sheet

C. IODINE TEST FOR POLYSACCHARIDES

Place 5-6 drops of glucose, fructose, sucrose, lactose, starch, H2O and
unknown in separate wells in your well plate.
Add 1 drop of iodine reagent to each.
A color change from yellow/brown to dark blue/black color is a positive test for
amylose in starch.
Record
D. HYDROLYSIS OF DISACCHARIDES AND POLYSACCHARIDES
Materials: 10% HCl, 10% NaOH, sucrose, starch Benedicts and Iodine Test solutions
Place 7-8 drops of 1% starch in two micro-centrifuge tubes and 7-8 drops of
sucrose in two more micro-centrifuge tubes.
To one sample of sucrose and starch, add 3-4 drops of 10% HCl. To the other
samples of sucrose and starch, add 3-4 drops of DI water. Label the test tubes
and gently shake the closed tubes to mix, and then heat in a boiling water bath
for 10 minutes.
Remove the test tubes from the water bath and let them cool.
To the samples containing HCl, add 10% NaOH (about 3-4 drops) to nand shake
the closed tube to mix.
Iodine Test: Place 5-6 drops of each solution in a well and add 1 drop of iodine solution
to each. Add extra iodine solution if the color disappears. Continue to add iodine
solution one drop at a time until a permanent color is obtained. Record your
observations. Determine if hydrolysis has occurred in each.
Benedicts Test: Add 9 drops of Benedicts reagent to each of the remaining samples
and heat in a boiling water bath for 3-4 minutes. Determine if hydrolysis has occurred in
each.
E. TESTING FOODS FOR CARBOHYDRATES

Takeyourtwofooditemandpreparethemforallthreetests
Takeasmallpieceandgrinditupwithmortarandpestle,adddistilledwaterto
createafairlydilutepastthatallowsyoutoseethecolorchangeforBenedicts/
Seligwanoff/Iodine
Fruit/veggiestarchtest:cutathinsliceoffandplaceonpapertowel,put12
Iodinedropsdirectlyontheslice
Cleanup:Scrubmortarandpestlewithwater/soap/brushanddry.

IV.

DATA REPORT SHEET

PART A, B, C IDENTIFYING CARBS

Indicate the color change in the table below. Example: blue orange
Carb
A. Benedicts
B. Seligwanoffs

C. Iodine

Glucose
Fructose
Lactose
Sucrose

Starch
Unknown
D. HYDROLYSIS OF DISACCHARIDES AND POLYSACCHARIDES
Indicate the color change in the table below. Example: blue orange
Sucrose + H2O Sucrose + HCl
Starch + H2O
Iodine

Starch + H2O

Benedicts
E. TESTING FOODS FOR CARBOHYDRATES

Whichtwofooditemsareyouplanningtotest?___________________________
Basedonyourknowledgeaboutcarbohydrateshypothesizewhatyourfooditems
willmostlikelytestpositivefor?
________________________________________________
Do the Tests: Indicate the color change in the table below. Example: blue orange

Test
Benedicts
Seligwanoffs
Iodine
V.

Food1:________________

Food2:______________

CONCLUSION/ DISCUSSION

Parts A, B, C. IDENTIFYING CARBS

Use the results table from A, B, C and identify if the following carbs where positive (write a
big + into the respective box)for Reducing Sugar, Ketose, Starch if negative leave empty
Carb
A. Reducing
B. Ketose
C. Starch
carb
Glucose
Fructose
Lactose
Sucrose
Starch
Unknown
Part D. HYDROLYSIS
Use the results table from A, B, C and identify if the following carbs where positive (write a
big + into the respective box)for Reducing Sugar, Ketose, Starch if negative leave empty
Sucrose + H2O Sucrose + HCl
Starch + H2O
Starch + H2O
Positive for

Starch
Positive for
Reducing sugar
Part E. FOOD TESTING
Use the results table from A, B, C and identify if the following carbs where positive (write a
big + into the respective box)for Reducing Sugar, Ketose, Starch if negative leave empty

Test
Reducingsugar
Ketose
Starch

Food1:________________

Food2:______________

Howdidyourhypothesisholdup?

Whatwassurprising/difficultaboutthefooditemtesting

Whatdidyoulikemostaboutthatlab?