CARBOHYDRATES

MISS I.N. EZECHUKWU

 REDUCING AND
NON-REDUCING SUAGRS
• A Reducing Sugar is any sugar that is capable of acting as a reducing agent
because it either has:
• A Free Aldehyde Group
• A Free Ketone Group
• A Hemiacetal Linkage

• The prescence of any of these groups allow the carbohydrate to undergo

easy oxidation, i.e the carbonyl carbon of the sugar is oxidized to a
carboxyl group.

• If the sugar gets oxidized, it causes reduction in another compound, hence

its name ''Reducing Sugar''.
• Non-Reducing Sugars are carbohydrates in which there is no free/available
Carbon atoms i.e there is no '-OH' group attached to the anomeric carbon, so
they cant reduce other compounds.

• In basic aqueous medium, they don’t generate any compounds containing an

aldehyde group. This is because they are not oxidized by weak oxidizing
agents i.e they are stable with respect to hydrolysis by bases.
• All Monosaccharides are Reducing Sugars e.g. Glucose, Fructose, Galactose

• The monosaccharides can be divided into 2 groups:

The Aldoses, which have an aldehyde group
The Ketoses, which have a ketone group

• The cyclic hemiacetal forms of Aldoses can open to reveal an Aldehyde group.

• Ketoses must 1st undergo Isomerization through a series of Tautomeric Shifts

to become Aldoses, i.e produce an aldehyde group, before they can act as
Reducing Sugars.
• Disaccharides can either be Reducing or Non-Reducing.

• Reducing Disaccharides like Lactose & Maltose have only one of their
anomeric carbons involved in the glysosidic bond, meaning they can convert to
an open chain form with an aldehyde group.

• Non-Reducing Disaccharides like Sucrose have glycosidic bonds between their

anomeric carbons, thus they can’t convert to an open chain form with an
aldehyde group; they are stuck in the cyclic form.
• Several qualitative tests are used to detect the presence of Reducing Sugars.
However, Non-Reducing Sugars don’t react with any of the Reducing Sugar
Test solutions.

• However, a non-reducing sugar can be hydrolysed using Dilute Hydrochloric

Acid in order to release its Reducing Sugar components, which can react
positively with the qualitative tests.

• METHOD: 6ml of 1% Sucrose in a test tube + 2 drops of Concentrated HCl.

Heat the tube in a boiling water bath for 15 minutes. Then test for presence of
Reducing Sugar using qualitative tests.
COMMON CHEMICAL REACTIONS OF
CARBOHYDRATES
1. DEHYDRATION
• When mixed with Concentrated Sulphuric Acid, in the presence of heat,
Carbohydrates undergo dehydration with the elimation of 3 Water
Molecules.

• During dehydration, Carbohydrates give out a Furfural or its derivative.

• The Furfural or its derivative formed condenses with α-Napitol to form

colored products.
MOLISCH TEST
• A general test for all Carbohydrates. Monosaccharides give a rapid
positive test. Disaccharides and polysaccharides react slower.

• The molisch reagent

dehydrates pentoses to
form furfural.

• The molisch reagent

dehydrates hexoses to
form 5-
hydroxymethylfurfural.
• The furfurals further react with
α-naphtol present in the test
reagent to produce a PURPLE
product.
2. REDUCTION OF CARBONYL GROUP
Sugars are classified as:
• Reducing Sugars
• Non-Reducing Sugars

• Under Alkaline conditions, Reduing Sugars tautomerize to Enediol. They then reduce
Cu2+ to Cu+ , which forms a RED precipitate: Copper (I) Oxide.
FEHLING’S TEST
• In an Alkaline medium, Reducing Sugars reduce Copper ions present in
Fehling's Solution to give an ORANGE to RED precipitate.

• Positive Result is detected by Reduction of the DEEP BLUE solution to a

RED precipitate. For a Negative Result, the solution will REMAIN BLUE.

• Sucrose doesn’t react with Fehling's reagent as it is a Non-Reducing Sugar.

BENEDICTS TEST
• Used to differentiate between Reducing and Non-Reducing Sugars i.e. Used to
detect the presence of Reducing Sugars.

• More Stable Reagent than Fehling's Reagent

• Benedict's Reagent contains BLUE Copper (II) Sulphate/CuSo4.5H2O, which is

reduced to RED Copper (I) Oxide by Aldehydes, thus oxidizing the Aldehydes to
Carboxylic acid.

• The Copper (I) oxide is insoluble in water, and so precipitates. The colour of the
final solution ranges from GREEN to BRICK RED depending on how many of the
copper (II) ions are present. The formation of a precipitate indicates a positive
result.
BARFOID’S TEST
• 1ml of the tested solution + 2ml of Barfoid Reagent
• Place test tubes into a boiling water bath and heat for 2 minutes. Then remove
& allow to cool.
• Formation of a GREEN, RED or YELLOW precipitate is a POSITIVE TEST for
REDUCING MONOSACCHARIDES.
• DO NOT HEAT THE TUBE FOR LONGER THAN 3 MINUTES as a FALSE POSITIVE
result can be obtained with Disaccharides if heated long enough.
OSAZONE TEST
• When Reducing Sugar is heated with PHENYLHYDRAZINE, Yellow Crystalline Compounds called
OSAZONES are formed.

• The 1st 2 Carbon atoms of a compound are involved in Osazone Formation.

• Osazone of different sugars are identified from their Crystalline forms, Time required for their formation
& their Melting points i.e.
Monosaccharides give NEEDLE-SHAPED/BROOMSTICK Crystals
Maltose gives SUNFLOWER-SHAPED Crystals
Lactose gives POWDER-PUFF Crystals
CHARCTERISTICS OF OSAZONES
• Osazones are INSOLUBLE

• Osazones have Characteristic Shapes & Melting Points

• Because both Carbons 1 & 2 are involved in the reaction, C-2 Epimers produce
the SAME OSAZONE

• Ketoses with configurations identical to Aldoses below C-2 give the SAME
OSAZSONES e.g. Glucose and Fructose
3. REDUCTION
4. OXIDATION
5. SELIWANOFF’S TEST
• This test distinguishes between Aldose and Ketose Sugars. When heated,
Ketoses are more rapidly dehydrated than Aldoses.

• The test reagent dehydrates Ketohexoses to form 5-hydroxymethylfurfural.

5-hydroxymethylfurfural further reacts with Resorcinol present in the test
reagent to produce a RED product within 2 minutes.

• Aldohexoses may react slightly/more slowly to produce a FAINT PINK colour.

6. IODINE TEST
• This is a qualitative test for the Detection of Polysaccharides.

METHOD: 1/2 ml of the Polysaccharide solution + 1 drop of Iodine solution.

If colour of the solution after addition of Iodine remains YELLOW, it is a
NEGATIVE result.
 COMPLEX
POLYSACCHARIDES
 POLYSACCHARIDES:
• Are Carbohydrates of higher molecular weight
• Contains very long chain of hundreds & thousands of Monosaccharides units
• They can be Straight or Branched
• They are made of 1 or different types of sugar
• All Monomeric units are linked together by GLYCOSIDIC LINKAGE
• Are mostly INSOLUBLE IN WATER
Polysaccharides are classified into 2:
• HOMOPOLYSACCHARIDES/HOMOGLYCANS: These are polysaccharides made up of
only ONE TYPE OF MONOSACCHARIDE or MONOSACCHARIDE DERIVATIVE. On
hydrolysis, they give only one type of monosaccharide. Eg Starch, Cellulose, Glycogen.
• HETEROPOLYSACCHARIDES/HETEROGLYCANS: These are polysaccharides made
up of MORE THAN ONE TYPE OF MONOSACCHARIDE or MONOSACCHARIDE
DERIVATIVE. On hydrolysis, they give 2 or many types of monomers. Eg Agar,
Agarose, Glycosaminoglycans/GAGs.
HETEROPOLYSACCHARIDES
AGAR & AGAROSE
• This is made up of D-Galactose & 3,6-anhydro L-Galactose

• Made of 2 Components: Agarose (Unbranched) & Agaropectin (Branched)

• It is dissolved in water at 100℃, which upon cooling sets into a gel.

• Agar cannot be digested by bacteria, so it’s widely used as a Supporting Media to Culture
Bacterial Colonies.

• It is also used as a Supporting Agent for Immuno-Diffusion and Immuno-Electrophoresis.

MUCOPOLYSACCHARIDES
• Also known as Glycosaminoglycans/GAGs
• These are Long, Unbranched Heteropolysaccharides made of repeating
Disaccharide units containing Uronic Acid & Amino Sugars.
• The Amino Sugar could be Glucosamine or Galactosamine
• The Uronic Acid could be D-Glucuronic acid or L-Iduronic acid
• The presence of Acetylated Amino Sugars, Sulfate & Carboxyl groups makes
them Acidic.
• Mucopolysaccharides are sometimes found in combination with protein to
form Mucoproteins/Proteoglycans.
• GAGs are the Major Gel Forming Components of The Extracellular Matrix of
Connective Tissue i.e. the anionic groups (Carboxy & Sulfate groups) being
Strongly Hydrophilic tend to bind large amounts of water to produce the gel-
like Matrix.

• The Heteropolysaccharide chains of GAGs repel one another & thus exist in
extended conformation in solutions. This produces the slippery consistency
of Mucus secretions & Synovial Fluid in joints.
BLOOD GROUP SUBSTANCES (BLOOD GROUP ANTIGENS)
• Red blood cell membranes contain several antigenic substances, on which
they are classified into different blood groups.

• These substances contain carbohydrates as glycoproteins or glycolipids.

Common examples are N-Acetylgalactosamine, Galactose, Fructose & Sialic
acid.
 HYALURONIC ACID
• It is a Sulphate-Free GAG
• Found in Synovial fluid of joints, Connective tissues & Cartilage
FUNCTIONS:
• Serves as a Lubricant & Shock Absorbant in joints
• Acts as sieves in the extracellular matrix
• Permits Cell Migration during Morphogenesis & Wound repair
 CHONDROITIN 4-SULFATE
• Most Abundant GAG
• Widely distributed in Bone, Cartilage & Tendons
FUNCTIONS:
• In Cartilage, it binds Collagen & holds fibers together in a tight strong network
• Role in Compressibility of Cartilage in Weight Bearing
 DERMATAN SULFATE
• Widely distributed in Skin, Cardiac Valves & Tendons
FUNCTIONS:
• Present in Sclera of eye where it aids in maintaining the overall shape of the eye
 HEPARIN
• Only Intracellular GAG
• Present in Granules of Mast Cells. Present also in the Lung, Liver & Skin
• Strongly Acidic due to presence of more Sulfate groups
FUNCTIONS:
• It is an Anticoagulant (Prevents Blood Clotting)
• Helps in the release of the enzyme Lipoprotein Lipase/LPL, which helps to clear lipidemia
after a fatty meal.
HEPARIN SULFATE
• This is an extracellular GAG found in basement membranes & is an essential
component of cell surfaces.

• It is Structurally Similar to Heparin but has a Lower Molecular Weight,

contains More Acetyl Groups & Less Sulfate Groups.

• It Determines Charge Selectiveness of the Renal Glomerulus.

THE END!!!