Carbohydrates

Background
• Carbohydrate are organic biomolecule that have chemical formula Cn (H2O) n

• Carbohydrates are widely distributed in plants and animals

• They have important structural and metabolic roles

Eg: Glucose- important carbohydrate
metabolic fuel of mammals
precursor for synthesis of some other carbohydrates in the body

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Classification of Carbohydrates

• Presence of aldehyde or ketone functional group classifies them as

• Aldoses
• ketoses

• May be classified as trioses, tetroses, pentoses, hexoses or heptoses

depending upon the number of carbon atoms

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Classification of Carbohydrates
• Monosaccharides-the simplest form of carbohydrates
• Cannot be hydrolysed into simplier carbohydrates
• Disaccharides –condensation products of two monosaccharide units
• Eg: maltose and sucrose
• Oligosaccharides-condensation products of three to ten
monosaccharides
• Polysaccharides-condensation products of more than ten
monosaccharide units eg: cellulose, glycogen, chitin, starch and
dextrins
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Monosaccharides of Biological Significance

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Disaccharides of Biological Significance

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Reducing and non reducing sugars
• Sugars can be classified as reducing or non-reducing; this classification is
dependent on their ability to donate electrons
• Reducing sugars can donate electrons (the carbonyl group becomes oxidised),
the sugars become the reducing agent
• Thus reducing sugars can be detected using the Benedict’s test as they reduce the
soluble copper sulphate to insoluble brick-red copper oxide
• Examples: glucose, fructose, maltose
• Non-reducing sugars cannot donate electrons, their electrons have been used in
the formation of the glycosidic bonds, therefore they cannot be oxidized.
• To be detected non-reducing sugars must first be hydrolyzed to break the disaccharide
into its two monosaccharides before a Benedict’s test can be carried out
• Example: sucrose
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Reducing and non reducing sugars
• A sugar that serves as a reducing
agent due to its free aldehyde
or ketone functional groups in its
molecular structure.
• Examples:
• glucose,
• fructose,
• lactose,
• arabinose and maltose

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Qualitative Test for carbohydrates
• Important test for detecting and classifying carbohydrates

• Qualitative analysis of carbohydrates is detected based on

the reagent’s utilization and the reaction between the test
sample and reagent.

• The reaction of the test sample with the chemical reagent

gives a significant colour, through which we can detect the
presence or absence of carbohydrates.
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Molisch’s Test
• General test for carbohydrates
• Reagent used: Naphthol in 95% Ethanol+ Conc
H2SO4
• Reaction:
• The conc sulphuric acid dehydrates the
carbohydrates to give furfural (Pentose) and 5-
Hydroxy methyl furfural in the case of hexoses.
• The fural derivatives condense with alpha naphthol
to form a violet-coloured precipitate(purple ring)
which is observed at the interface of the two
solutions which is the positive result.

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Iodine test
• This is a tests for polysaccharides particularly starch

• Reagent used: Lugol’s Iodine(Potassium Iodide(KI) in Iodide(I)

solution)

• Principle:
• Iodine forms coordinate complexes between the helically coiled
polysaccharide chain which causes the change of the original brownish
colour to blue black/deep blue colour indicting a positive result

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Iodine Test observation

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Benedict’s Test
• This test can be used for the identification of reducing and non
reducing sugars
• Reagent used: Benedict’s reagent-Copper sulphate,
Sodium Carbonate
Sodium Citrate

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 Benedict’s Test
• Principle:
• Reducing sugars under alkaline conditions
tautomerise-structural isomers and form enediols
• Enediols- are powerful reducing agents
• They reduce Copper sulphate which produce Cupric
ions(Cu ) to form cuprous ions which form cuprous
2+

hydroxide with hydroxide ions in solution to form red

cuprous oxide upon heating

Control Reducing
-ve result sugar +ve
result
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Fehling’s test
• This test can be used for the identification of reducing and non
reducing sugars

• Reagent used: Fehling’s Solution- It is made fresh by combining equal

volumes of solutions A and B
• Solution A-Copper sulphate,
• Solution B- Potassium Sodium Tartrate tetrahydrate
Sodium hydroxide

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Fehling’s test
Principle
• The aldehyde and ketone group on the carbohydrate are involved in
the reaction
• When the reaction is completed, the copper II ions are reduced to
Copper I oxide, which forms a red precipitate and is insoluble in water.
A positive test result is indicated by the presence of this red
precipitate

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Procedure-

Description of the procedure of fehling’s test

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Barfoed’s Test
• This test is also for the identification reducing monosaccharides from
disaccharides by controlling pH and time of heating.
NB: Monosaccharides react very fast and disaccharides react very slowly
• Reagent used: Barfoed’s reagent-Copper acetate in distilled water with
1.8ml of Glacial acetic acid
• Procedure: 1ml test sample + 2ml Barfoed’s reagent+ boil for 5-30
minutes.
• Result: there is the formation of cuprous oxide hence the brick red colour
observed within 3 minutes is the positive result for the monosaccharide.
on prolonged heating disaccharides also give a positive result.

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Barfoed’s Test

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Seliwanoff’s test
• This test is used to differentiate between sugars that have a ketone
group (ketose) and sugars that have an aldehyde group (aldoses).
• It is a timed color reaction specific to ketohexoses.
• Reagent used: Seliwanoff’s reagent-(Resorcinol and concentrated
HCl)

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Seliwanoff’s test
• Principle:
• Ketoses are more rapidly dehydrated than aldoses.
• Ketoses undergo dehydration in the presence of
concentrated acid to yield 5-hydroxymethyl
furfural.
• The dehydrated ketose reacts with two equivalents
of resorcinol in a series of condensation reactions to
produce a complex (not a precipitate), termed -ve result + ve result
Cherry red-coloured
xanthenoid, with deep cherry red color. complex formed

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Anthrone Test
• This test is used to detect the presence of carbohydrates in a given
solution.
• Reagent used: Anthrone in concentrated Sulphuric acid
• The concentrated acid catalyzes the dehydration of the
monosaccharides to form furfural (from pentoses) or hydroxyl furfural
(from hexoses). The furfural or hydroxyl furfural formed condenses
with two molecules of naphthol from the Anthrone reagent to form a
blue-green complex.

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Anthrone Test

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Questions

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