Experiment: Carbohydrates (Part 1)-
Molisch Test
The Monosaccharides
Carbohydrates
- Include:
• polyhydroxy aldehyde
• ketones
• their by-products
- make up most of our diet
- serves as the major source of
energy
- role: metabolic processes
• furnishing carbon chain for
compound synthesis
Categories:
• monosaccharides
• disaccharides
• polysaccharides
Monosaccharides
- simple sugars
• highly soluble in water
• less soluble in ethanol
• insoluble in ether
- can’t hydrolyzed to simpler units
- either aldoses or ketroses
Classifications:
• tetroses
• pentoses
• hexoses
= depending on the # of carbon
atoms they possess
- exhibit mutarotation
• can exist in a- and B-forms
Ability of Chemical Tests used to
detect presence of carbs:
• form furfural and its derivatives
• reduce and form characteristic
compounds with reagents (Tollen’s
and Benedict’s)
1.
- general test for carbs
- sugars mixed with a-naphthol
- there’s formation of a lower layer
of acid due to the conc. H2SO4,
conc. sulfuric acid will dehydrate
the sugar and react with the
alcohol forming furfural or
hydroxymethyl furfural
- presence of carbs: purple ring
2. Seliwanoff’s Test
- used to differentiate ketohexoses
from aldohexoses
- Ketohexoses react faster with
solution containing hydrochloric
acid and resorcinol than
aldohexoses
• dehydrated ketohexoses: form
bright cherry red condensation
product
• aldohexose: yield pale pink
coloration (neg. result)
- Prolonged heating of samples
should be avoided
3. Phenylhydrazine Test (Osazone
formation)
- yellow ring: formed when sugar is
added to phenylhydrazine & sodium
acetate (heated)
• precipitate formed may be
compared with standards using
difference in melting points
(to identify the composition of
the precipitate)
- Crystalline Structure
• Glucosazones: yellow needles
aggregated “bundles of hay”
• Glucose, maltose, mannose form
the same osazones
• Lactosazone crystals: regular
clusters of fine needles,
“powder puff”
• Maltosazone Crystals: “star-
shaped”
Experiment: Carbohydrates
Mono and disaccharide groups
- possess reducing properties (groups
containting the potentially free
aldehyde or ketone group)
- reduce alkaline metals
- transformed into organic acids
Solutions most commonly employed for
this purpose:
• alkaline solution of copper
sulfate(Trommer’s, Fehling’s &
Benedict’s reagent)
• alkaline sol. of Bismuth subnitrate
(Nylander’s rgnt)
- in acid solution, reducing
monosaccharide also exert action
- sol. of cupric acetate in weak acetic
acid (Barfoed’s rgt) is reduced by
monosaccharied but not by
disaccharides
Tests:
1. Benedict’s Test
- very sensitive test done under mildly
alkaline conditions
- contains CuSO4, Na2CO3 & sodium
citrate
- postive: brick red precipitate of
Cu2SO4
- most aldehydes have the ability to
reduce Benedict’s rgt.
- Other compounds yield postive result:
formic acid, hydrazobenzene, phenols,
pheylhydrazine, pyrogallol, uric acid
2. Barfoed’s Test
- Barfoed’s rgt contains cupric acetate
• oxidizes monosaccharides but not
oligosaccharides
- used to distinguished between
monosaccharides, disaccharides and
oligosaccharides
- Disaccharides are less easily
oxidized but are oxidized if they
undergo prolonged heating causing
hydrolysis (positive result)
- the use of a more conc. disaccharide
solution may give a faster reaction
that that of a relatively more dilute
monosaccharide solution
- this test is carried out under acidic
rather than basic medium
3. Fehling’s Test
- used extensively in carbohdyrate
chemistry
- positive: brick red precipitate
- aldoses are easily oxidized to yield
carboxylic acids
- cupric ion complexed with tartate ion
is reduced to cuprous oxide
Sucrose
- does not react with Fehling’s rgt
- disaccharide of glucose and fructose
- non-reducing sugar
Glucose
- reacts with Fehling’s rgt to form an
orange to red ppt
- reducing sugar
Fructose
- gives a positive test with Fehling’s
solution
• fructose is converted to glucose
and mannose under alkaline
conditions
• can be explained by the keto-enol
tautomerism
4. Trommer’s Test
- very much like benedict’s test for
reducing sugar
- produce oxides of copper that
precipitate out of solution (color
change)