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Laboratory Manual
INTRODUCTION
This activity on carbohydrates will involve two parts: its common and specific reactions.
It will entail two two-hour laboratory periods.
Common Reactions
Carbohydrates are classified according to the complexity of their molecules. The simplest
group, called monosaccharides, are the building blocks from which higher sugars,
(dissacharides) and more complex groups (polysaccharides) are formed. The
monosaccharides are sub-classified according to the number of carbon atoms in their
unit chains into trioses, tetroses, pentoses and hexoses; and according to the position of
their carbonyl groups into aldoses and ketoses. Thus, they are sometimes designated as
aldopentoses and ketohexoses. The ending ose is generally used in naming
carbohydrates and the term sugar refers to those possessing sweet taste, most of which
are monosaccharides and dissacharides.
Only a few classes of carbohydrates have special biological importance. These include
the hexoses: glucose, also known as dextrose or grape sugar, fructose or levulose, and
galactose; and those that are compounds of hexoses, the hexosans.
Glucose is a normal constituent of the blood and tissue fluids of the human body. D-
galactose, also called brain sugar, is found as a cerebroside in the brain and as a
ganglioside in the nerve tissues. It is also the sugar of galactolipids. A glucose
derivative, glucosamine with protein form glycoproteins which are involved in many vital
functions in the body. Certain pentoses are also important. Ribose and deoxyribose are
constituents of nucleic acids and are also present in some enzymes. Trioses and related
oxidation and/or reduction products are involved in the oxidation and synthesis
mechanisms in the body. Structurally, carbohydrates are polyhydroxyketones.
Accordingly, their chemical reactions are characteristic of both the OH and C=O
functional groups.
H
O H O
H OH
-3H2O
H OH H O
H OH conc. H2SO4
CH2 OH H H
D - ribose furaldehyde or furfural
Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga
University, Zamboanga City, Philippines.
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Laboratory Manual
O
H O
H OH -3H2O
HO H
conc. H2SO4 H O
H OH
H OH
H CH 2 OH
OH
D - glucose 5-hydroxymethylfurfural
With concentrated sulfuric acid, H2SO4, the polysaccharides and dissacharides are first
hydrolyzed to monosaccharides before they are dehydrated. Pentoses yield furfural
whereas hexoses yield hydroxymethylfurfural which is further degraded into levulinic
acid.
The disaccharide sucrose is non-reducing because both rings of glucose and fructose are
in the acetal or ketal form. So they cannot be in equilibrium with their aldehyde forms.
H O H OH H OH
H OH OH O
HO H OH- HO H OH-
HO H
H OH H OH H OH
H OH H OH H OH
CH 2 OH CH 2 OH CH 2 OH
glucose 1,2-enediol fructose
OH-
H O
HO H
HO H
H OH
H OH
CH2OH
mannose
Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga
University, Zamboanga City, Philippines.
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Laboratory Manual
Ketoses like fructose give a positive reaction to tests for reducing sugars because of the
Lobry de Buryn-von Eckenstein transformation. The alkaline solution favors the
formation of the enediol and suppresses formation of the ring structure. The outline
reaction below shows the transformation of the hexoses into a common enediol form.
The outline shows the change only in the first 2 carbons of each sugar.
The enediols formed in alkaline medium dilute Ba(OH)2 decompose readily when heated.
The double bond of the enediol moves down the carbon chain giving rise to a variety of
isomers and eventually to the cleavage of the C=C chain.
H H
H O H OH H
H OH OH
H OH OH H OH
OH
HO H HO H HO
HO
H OH H OH OH
H OH
H OH H OH H OH
H OH
CH 2 OH CH 2 OH CH 2 OH
CH 2 OH
The double bond breaks between 3C and 4C; forming 2 trioses, glycerose and
dihydroxyacetone, which provide the pathway to the oxidation of glucose in the Krebs’s
cycle. In the absence of oxygen, the 3,4-enediol is predominant.
Other decomposition products when the double bond moves further down are
formaldehyde, glycoladehyde, lactic acid, methyl glyoxal and reductone. Some of these
fragments form complex resinous compounds reducing yellow to brownish color. The
property of enediols is the basis of Moore’s test for reducing sugars.
In the Tollen’s test, both aliphatic and aromatic aldehydes are oxidized to carboxylic
acids and the Ag ion is reduced to elemental Ag deposited on the sides of the tube.
O O
+
R + 2 Ag(NH 3)2 + 2 OH- 2 Ag + R
+
+ H2O + 3 NH3
H O NH 4
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University, Zamboanga City, Philippines.
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APPARATUS/MATERIALS CHEMICALS/REAGENTS
PROCEDURE
A. Molisch’s Test
B. Action of Alkali (DO NOT PERFORM. Just search for theoretical results.)
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C. Reduction Tests
Fehling’s Test
2. Take one of the test tubes, while heating on a water bath, add 1% glucose
solution drop by drop. Note the result.
3. Repeat the test on the other test tubes, using different 1% sugar solutions of
xylose, fructose, maltose, sucrose, starch, glycogen and an unknown sugar
solution.
Benedict’s Test
2. Repeat the test on 1% solutions of xylose, fructose, lactose, sucrose and starch.
Barfoed’s Test
Nylander’s Test (DO NOT PERFORM. Just search for theoretical results.)
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University, Zamboanga City, Philippines.
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Tollen’s Test
QUESTIONS
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Data
A. Molisch Test
1% xylose
1% lactose
1% sucrose
1% starch
1% furfural
Unknown
sugar
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1% fructose
1% galactose
1% 1% glucose
Ba(OH)2
1% fructose
1% galactose
C. Reduction Tests
1% xylose
1% fructose
1% lactose
1% maltose
1% sucrose
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University, Zamboanga City, Philippines.
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1% starch
1% glycogen
Unknown
Benedict’s 1% glucose
Test
1% xylose
1% fructose
1% lactose
1% sucrose
1% starch
Barfoed’s 1% glucose
Test
1% fructose
1% xylose
1% lactose
1% maltose
1% sucrose
Unknown
sugar
Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga
University, Zamboanga City, Philippines.
Chemistry Laboratory 204: Organic Chemistry and Biochemistry 10
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1% lactose
1% maltose
1% fructose
1% xylose
1% lactose
1% maltose
Tollen’s 1% glucose
Test
Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga
University, Zamboanga City, Philippines.