Lab Carbohydrates

Objective:
Study the various qualitative tests of carbohydrates.
Identify an unknown carbohydrate.
Background:
What follows are descriptions of how different test reagents react with different carbohydrates.
Aniline acetate is used to identify pentose and hexose sugars. If a sugar sample is heated with
Cl and yields a bright pink color with the addition of aniline acetate! the sugar is a pentose.
"his reactions yields #$furfuraldehyde. If a sugar sample is heated with Cl and yields a
negligible pink color with the addition of aniline acetate! the sugar is a hexose. "his reactions
yields hydroxymethylfurfural.
%
C
%

%
C
%

C
#
%
#$furfuraldehyde hydroxymethylfurfural
bright pink color negligible pink color
Barfoeds reagent is prepared by mixing acetic acid and copper&II' acetate. "he Cu
(#
ion acts
as a weak oxidi)ing agent on monosaccarides! but is too weak to oxide a disaccharide. "he
Cu
(#

is reduced to Cu
(
and forms copper&I' oxide! which is red. *emember that Cu
(#
is blue is
solution! so your reagents appearance will change from a clear light blue to a red precipitate
laden colorless solution.
Benedicts reagent is a combination of copper&II' sulfate! sodium citrate and sodium carbonate.
With the addition of the sodium carbonate! the mixture takes on a basic characteristic. "he
copper&II' ion in +enedict,s reagent is strong enough to oxidi)e monosaccarides and some
disaccharides. "his is the reagent used by medical test facilities to check for glucose in urine.
Fermentation, by the addition of yeast! can be use to as a general test for carbohydrates.
+ubbles of carbon dioxide are produced at different rates depending on the carbohydrate used.
Seliwanoffs reagent will differentiate between aldose and ketose monosaccarides. "he
reagent is a mix of Cl and resorcinol! shown below. -o change will occur testing aldose or
disaccharides! but there will be a color change with ketose carbohydrates. .etohexoses will
give a cherry red solution and ketopentose will give a blue$green solution.
%
%
resorcinol
rocedure:
/or most of the following sections of this procedure you will be testing several different known
carbohydrates! fructose! glucose! lactose! maltose! sucrose and xylose. 0ou will be given an
unknown. "he unknown will be one of these six know carbohydrates. 1sing these test! identify
your unknown carbohydrate. 0ou will also be testing two other carbohydrate solutions! honey
and 2ell$%

.
It would be prudent to do all steps in each part for all solutions at the same time. "his way you
can compare all the knowns to your unknown.
!"#"O$" %:&&&&
3art 45
4. +egin your testing with the fermentation as it will take about an hour to react.
#. 6dd 7mL of each of the solutions and the unknown to different test tubes.
7. 6dd 7mL of the yeast solution to each test tube! cover the tube with a piece of 3ara$/ilm!
invert to mix and remove 3ara$/ilm.
8. 3lace the test tubes in a warm water bath! 89$:9C. ;o not over heat! the yeast will die.
:. *ecord your observations over an hour.
3art #5
4. Concentrated Cl is highly volatile and noxious! perform this part under the hood.
#. 6dd #mL of the xylose solution to a 4#:mL flask.
7. 6dd 49mL of deioni)ed! ;I! water to flask.
8. 6dd 49mL of Concentrated Cl to flask.
:. +oil solution for 4 minute.
<. *emove from heat and place a piece of filter paper over the flask.
=. ;rip 7 drops of aniline acetate on the section of the paper directly over the mouth of the
flask.
>. *ecord observations.
?. *epeat using your unknown.
3art 75
4. -ext! the +arfoed,s test.
#. 6dd 4mL &#9 drops' of each of the solutions and the unknown to different test tubes.
7. 6dd 4mL &#9 drops' of the +arfoed,s reagent to each test tube and invert.
4. 3lace the test tubes in a boiling water bath! and make observations over the next :$<
minutes.
3art 85
8. "he +enedict,s test will be next.
:. 6dd 4mL &#9 drops' of each of the solutions and the unknown to different test tubes.
<. 6dd 4mL &#9 drops' of the +enedict,s reagent to each test tube.
=. 1se a piece of 3ara$/ilm and shake tub thoroughly with your thumb over the tube mouth.
>. 3lace the test tubes in a boiling water bath at the same time.
?. "he bath will stop boiling. 6llow the tubes to sit in bath until # minutes after the water begins
to boil again.
49. *ecord your observations.
3art :5
4. "he Seliwanoff,s test is the final analysis
#. 6dd 4 drop of each of the solutions and the unknown to different test tubes.
7. 6dd #mL of the Seliwanoff,s reagent to each test tube.
8. 1se a piece of 3ara$/ilm and shake tub thoroughly with your thumb over the tube mouth.
:. 3lace the test tubes in a boiling water bath at the same time.
<. "he bath will stop boiling. 6llow the tubes to sit in bath until 8 minutes after the water begins
to boil again.
=. *ecord your observations after 4 minute and after 8 minutes.
'oncluding (uestions:
4. Which unknown where you given@
#. What type&s' of carbohydrates are gelatin and honey@ Aono or disaccharides! ketose or
aldose@
7. What products are generated by the yeast en)yme fermentation of fruit sugar@ Write out this
chemical equation.
8. When in a +enedict,s and +arfoed,s test the solutions changes from a clear blue liquid to a
redBbrown ppt! what is occurring@
a. What is blue! what is red@
b. What caused the change@