BOOKS RECOMMENDED: TEXT BOOKS

• Biochemistry for Medical students by [Link] & Shree Kumari.

• Medical Biochemistry by M.N. Chatterjea and Rana Shinde.
• Text Book of Medical Biochemistry by Ramakrishnan, Prasannan & Rajan.
• Medical Biochemistry by Debajyoti Das.
• Biochemistry by [Link].
• Medical Biochemistry - [Link].

• REFERENCE BOOKS:
• Harper's Biochemistry.
• Medical Biochemistry by [Link].
• Biochemistry by [Link].
• Biochemistry by Orten & Neuhans.
Carbohydrate Chemistry

DR. PANKAJ KUMAR GUPTA

(MBBS, MD, DA, PGDCR, PGDHHM, PGDMLS),
Associate Professor, Dept. of Biochemistry,
AIMMMCR, BHILAI
Learning Objectives
Define carbohydrates. Enumerate the classification of carbohydrates
with examples and discuss the functions of carbohydrate
Discuss monosaccharides giving examples as energy fuel. Describe
biologically important hexose derivatives
Describe the formation of glycosides and its therapeutic importance
Discuss oligosaccharides and disaccharides, giving their importance
and examples
Discuss polysaccharides with examples as a storage form like
glycogen, structural elements like glycosaminoglycans in the human
body and explain what is meant by the terms resistant starch,
glycemic index, and dietary fiber.
The carbohydrates are widely distributed both in animal and plant
tissues.
Chemically, they contain elements carbon, hydrogen & oxygen
Empirical formula of many simple carbohydrates is [CH2O]n. Hence,
the name “carbohydrate”, i.e. hydrated carbon.
They are also called “saccharides”. In Greek, saccharon means sugar.
Definition
Carbohydrates are aldehyde or ketone derivatives of polyhydroxy
alcohols or as compounds that yield these derivatives on hydrolysis.
 Classification of Carbohydrate
1. Monosaccharides
2. Oligosaccharides
3. Polysaccharides.
• Monosaccharides (Greek: Mono = one)
• Monosaccharides are also called simple sugars.
• The term sugar is applied to carbohydrates that are soluble in water
and sweet to taste.
• They cannot be hydrolyzed into a simpler form.
• They may be subdivided into two groups i.e Aldose and ketose.
Oligosaccharides (Greek: oligo = few)
Short chain of monosac­charide units (2 to 10 units), joined together
by glycosidic bond
On hydrolysis, gives two to ten molecules of simple sugar
(monosaccharide) units.
 Oligo­saccharides are subdivided into different groups based on the
number of monosaccharide units present
The disaccharides which have two monosaccha­ride units are the most
abundant in nature.
Oligosaccharides with more than three subunits are usually found in
glycoproteins; such as blood group antigens.
 Polysaccharides (Greek: Poly = many) or Glycans
Polymers consisting of hundreds or thousands of monosaccharide
units.
They are also called glycans or complex carbohydrates.
They may be either linear, (e.g. cellulose) or branched, (e.g. glycogen)
in structure.
Polysaccharides have high molecular weight and are only sparingly
soluble in water.
They are not sweetish and do not exhibit any of the properties of
aldehyde or ketone group.
Polysaccharides are of two types.
Homopolysaccharides (homoglycans)
Heteropolysaccharides (heteroglycans).
Homopolysaccharides (Homoglycans)
When a polysaccharide is made up of several units of one and the same
type of monosaccharide unit, it is called homopolysaccharide.
The most common homoglycans are:
– Starch
– Dextrins
– Glycogen
– Inulin
– Cellulose.
Some homopolysaccharides serve as a storage form of
monosaccharides used as fuel, e.g. starch and glycogen, while others
serve as structural elements in plants, e.g. cellulose.
Heteropolysaccharides (Heteroglycans)
They contain two or more different types of monosaccharide units or
their derivatives.
Heteropolysaccharide present in human beings is glycosaminoglycans
e.g.
– Heparin
– Chondriotin sulfate
– Hyaluronic acid
– Dermatan sulfate
– Keratan sulfate
– Blood group polysaccharides.
Figure 3.1: Classification of polysaccharide.
 Functions of Carbohydrates
 Source of energy for living beings, e.g. glucose
 Storage form of energy, e.g. glycogen in animal tissue and starch in
plants
 Serve as structural component, e.g. glycosamino­glycans in human,
cellulose in plants and chitin in insects
 Non-digestible carbohydrates like cellulose, agar, gum and pectin
serve as dietary fibers.
 Constituent of RNA and DNA, e.g. ribose and deoxyribose sugar.
 Carbohydrate play a role in lubrication, cellular intercommunication
and immunity.
 Carbohydrates are also involved in detoxification, e.g. glucuronic
acid
 Structure of Glucose
[Link] straight chain structural formula (Fisher projection)
[Link] formula (Ring structure or Haworth projec­tion)
 Monosaccharide in solution mainly present in ring form.
 In solution, the aldehyde group of glucose at C-1 reacts with alcohol
(OH) group of C-5 to form six membered ring called glucopyranose
 The aldehyde group of glucose at C-1 reacts with alcohol (OH) group
of C-4 to form five membered ring called glucofuranose.
 In case of glucose, the six membered glucopyranose is much more
stable than the glucofuranose ring.
 In the case of fructose, the more stable form is fructofuranose
 Isomerism/Stereoisomerism
The compounds possessing identical chemical (molecular) formula but
different structures are referred to as isomers.
Important types of isomerism
1. Aldose- ketose isomerism
2. D and L isomerism
3. Optical isomerism
4. Epimerism
5. Anomerism (α and β Anomerism)
Aldose-Ketose isomerism
D and L Isomerism
 D and L isomerism depends on the orientation of the H and OH
groups around the asymmetric carbon atom adjacent to the terminal
primary alcohol carbon.
 Carbon atom number 5 in glucose determines whether the sugar
belongs to D or L isomer.
 When OH group on this carbon atom is on the right, it belongs to D-
series, when it is on the left; it is the member of the L-series.
 The structures of D and L-glucose based on the reference
monosaccharide, D and L glyceraldehyde, a three carbon sugar.
D and L isomers (enantiomeric pairs) of glyceraldehyde
D and L isomers of glucose.
 Most of the monosaccharides in the living beings belong to the D-
series. (unlike amino acids which are L isomers).
 D and L isomers are mirror images of each other. These two forms are
called enantiomers.
Optical Isomerism
Optical activity is the capacity of a substance to rotate the plane polarized
light passing through it.
When a beam of plane-polarized light is passed through a solution of
an optical isomer, it will be rotated either to the right and is said to be
dextrorotatory (d) or (+) or to the left and is said to be, levorotatory (l)
or (-).
 In solution, glucose is dextrorotatory that is why glucose solutions
sometimes known as dextrose.)
 d-, and l- nomenclature is unrelated to D-and L-isomerism. This
nomenclature is obsolete.
When equal amount of (+) and (- ) isomers are present, the resulting
mixture has no optical activity such a mixture is said to be a racemic
mixture.
Epimerism
When two monosaccharides differ from each other in their
configuration around a single asymmetric carbon (other than anomeric
carbon) atom, they are referred to as epimers of each other.
Epimers of glucose.
Anomerism
Carbon-1 of glucose in the open chain form becomes an asymmetric
carbon in the ring form and two ring structures can be formed. These
are:
• α -D-glucose
• β -D-glucose
Generation of new asymmetric carbon atom at C1
α and β Anomers
 The designation α means that the hydroxyl group attached to C-1 is
below the plane of the ring, β means that it is above the plane of the
ring.
 The C-1 carbon is called the anomeric carbon atom and so, α and β
forms are anomers.
 Glycoside Formation
When the hydroxyl group on the anomeric carbon of a
monosaccharides reacts with OH or NH group of another compound (it
may or may not be a monosacc­haride) glycosides are formed.
The N type of glycoside bonds
 Therapeutic Importance of Glycosides
 Found in many drugs, e.g. in antibiotic streptomycin.
 Cardiac glycosides such as Ouabain and digoxin increase heart muscle
contraction and are used for treatment of congestive heart failure.
 Daunorubicin is used to treat leukaemia.
 Doxirubicin is used to treat cancers.
 Oligosaccharides
Oligosaccharides consist of short chains of mono­saccharide units (2
to 10 units)
Joined together by glycosidic linkages
Among oligosaccharides disaccharide is most abundant
DISACCHARIDES
Disaccharides consist of two monosaccharide units.
They are crystalline, water soluble and sweet to taste.
They are subclassified on the basis of the presence
or absence of free reducing group.
 In disaccharides the glycosidic linkage may be either α or β
depending on the configuration of the atom attached to the
anomeric carbon of the sugar
 Polysaccharides (Glycans)
Composed of ten or more monosaccharide units or their derivatives.
Monosaccharide units are joined together by glycosidic bond
Colloidal in nature.
Polysaccharides Subclassified into
1. Homopolysaccharide
2. Heteropolysaccharide
 Homopolysaccharides or Homoglycans
Starch
 It is the storage form of glucose in plants.
 Starch is composed of two constituents, amylose and amylopectin.
Amylose
Amylose is a linear polymer of D-glucose units joined by α-1, 4
glycosidic linkages
Amylopectin
Amylopectin is branched structure with α-1, 4 and α-1, 6 glycosidic
linkages
Amylose, glucose units joined by α-1,4 linkages
Structure of amylopectin
Amylopectin, glucose units joined by α-1,4 and α-1,6 linkages
Glycogen (Animal Starch)
Glycogen is the major storage form of carbohydrate in animals, found
mostly in liver and muscle. It is often called animal starch.
Functions
• The function of muscle glycogen is to act as a readily available source
of glucose for energy within muscle itself
• Liver glycogen is concerned with storage and maintenance of the
blood glucose.
Diagrammatic representation of glycogen molecule
Cellulose
 Constituent of cell wall of plants.
 Unbranched polymer of glucose linked by β-(1, 4) glycosidic linkages
and not α-(1,4)
 Humans lack an enzyme cellulase that can hydrolyze β-(1, 4)
glycosidic linkages of poly­saccharides, cellulose cannot be digested
and absorbed
Structure of cellulose
Importance Of Cellulose
 Cellulose is a component of fibre in the diet
 High fibre diet is associated with reduced incidence of
cardiovascular disease
colon cancer
diabetes
 It increases bulk of stool, aids intestinal motility and prevents
constipation.
Inulin
Inulin is a polymer of D-Fructose (Fructosans) linked together by β-
(1 → 2) glycosidic linkage.
It occurs in the tubers of some plants, e.g. chicory, bulb of onion
and garlic.
Inulin is not hydrolyzed by α-amylase but is hydrolyzed by inulinase,
which is not present in the humans and so it is not utilized as food.
Clinical importance of Inulin
Inulin has clinical importance as it is used in the studies of glomerular
filtration rates (kidney function test).
Glycosaminoglycans (GAGs) or Mucopolysaccharides
Glycosaminoglycans were first isolated from mucin which led to the
original naming of these compounds as mucopolysaccharide.
Structure of GAG
Unbranched, made up of repeating disaccharides :
(amino sugar : uronic acid)n
 Amino sugar : D-glucosamine or
D-galactosamine
 Uronic acid : Glucuronic acid or its epimer
Iduronic acid.
Sugar acids produced by oxidation of glucose
 Polymer of amino sugar - Uronic acid is attached covalently to
proteins called core protein to form proteoglycans.
 A resulting structure resembles a “bottle brush”.
Bottle brush structure of proteoglycan monomer
 The proteoglycan monomer associates with a molecule of hyaluronic
acid to form proteoglycan aggregates.
 The association is not covalent, but occurs through ionic interaction
between core protein and the hyaluronic acid.
 The association is stabilized by additional small proteins called link
proteins.
Figure: Proteoglycan aggregate.
Occurrence Of Gags
 Synovial fluid of joints
 Vitreous humour of the eye
 Arterial walls
 Bones and
 Cartilage.
Different types of GAGs
Hyaluronic Acid
Condroitin sulfate
Dermatan sulfate
Heparin
Heparin sulfate
Keratan sulfate
 With the exception of hyaluronic acid, all the GAGs contain sulfate
group.
 The amount of carbohydrates in proteoglycans is up to 95 percent
of its weight
Different types of GAGs differ from each other in:
 Uronic acid composition
 Amino sugar composition
 Linkage between amino sugar and uronic acid
 Chain length of the disaccharide polymer.
 Presence or, absence of sulfate groups
 Nature of core protein to which they are attached.
 Their tissue and subcellular distribution and their functions
Functions Of GAGs
 Structural component. They are major components of the extracellular
matrix or ground substance.
 Have special ability to bind large amounts of water, producing a gel
like matrix which functions as a cushion against mechanical shocks
 Negatively charged glycosaminoglycans chain repel each other, and
proteoglycans occupy a very large space and act as “molecular
sieves”, determining which substances to enter and leave cells.
 They also give elasticity (resilience) to cartilage, permitting
compression and re-expansion.
 They lubricate joints.
Strike a Note
 Glycosaminoglycans were first isolated from mucin which led to
original name as mucopolysaccharide.
 One component is always an amino sugar hence the name
glycosaminoglycans.
 Proteoglycans consist of up to 95% carbohydrate by weight.
 Except for hyaluronic acid, the glycosaminoglycans are linked to
protein.
 Glycosaminoglycan not covalently linked to protein is hyaluronic
acid.
 Glycosaminoglycan with no Uronic acid is keratan sulfate.
 Glycosaminoglycan with not sulfate group is hyaluronic acid.

 Glycosaminoglycans are generally extracellular, only intracellular

glycosaminoglycan is heparin.
 GLYCOPROTEIN
Proteins to which oligosaccharides are covalently attached.
Glycoproteins contain much shorter carbohydrate chain than
proteoglycans.
These oligosaccharide chains are often branched instead of linear and
may or may not be negatively charged as proteoglycans.
Functions Of Glycoproteins
 Almost all the plasma proteins of humans are glycoproteins, except
albumin.
 Many integral membrane proteins are glycoproteins.
 Most proteins, such as antibodies, hormones and coagulation factors
are glycoproteins
 Serve as lubricant and protective agent
 It also serve to transport molecules.
The oligosaccharide chains:

Glucose (Glu);
mannose (Man);
galactose (Gal);
N-acetyl glucosamine (GluNAc);
N-acetyl galactosamine (GalNAc);
L-fucose (Fuc) and
N-acetyl neuraminic acid (NANA).
Reaction of monosaccharides

In sugars, the following 3 properties will be seen together:

A. Mutarotation
B. Reducing property
C. Formation of osazone with phenylhydrazine.
Mutarotation.
When a fresh solution of D glucose is prepared, its specific
rotation of polarised light is +112o;
but after 12-18 hours it changes to +52.5o.
• This change in rotation with time is called mutarotation.
Explanation:

D-glucose has two anomers, alpha and beta varieties.

These anomers are produced by the spatial configuration with

reference to the first carbon atom in aldoses and second carbon atom
in ketoses.

Hence these carbon atoms are known as anomeric carbon atoms.

Alpha-D-glucose has specific rotation of +112 degree and
beta-D-glucose has +19 degree.

Both undergo mutarotation and at equilibrium one-third molecules

are alpha type and 2/3rd are beta variety.

to get the specific rotation of +52.5.

ENEDIOL FORMATION
In mild alkaline solutions, carbohydrates containing a
free sugar group (aldehyde or keto) will tautomerise
to form enediols.
In mild alkaline conditions, glucose is converted into
fructose and mannose.
The interconversion of sugars through a common
enediol form is called
Lobry de Bruyn-Van Ekenstein transformation.
 1,2-enediol form
Glucose Fructose
(intermediary)
BENEDICTS REACTION
Benedict's reagent is very commonly employed to detect the
presence of glucose in urine (glucosuria) and is a standard
laboratory test employed in diabetes mellitus.

Benedict's reagent
contains sodium carbonate, copper
sulphate and sodium citrate.
 Benedict's Reaction

Sugar——>enediol Cu++ ¬—CuSO4

Oxidised Reduced

Cu+ ®2Cu(OH)
Sugar acid ®Cu2O

Glucose is a Reducing sugar

Semi-quantitative test
Benedict’s solution

5 ml solution
+ 0.5 ml glucose
+ boil for 2 min
OSAZONE FORMATION

All reducing sugars will form osazones with excess of

phenylhydrazine when kept at boiling temperature.

Osazones are insoluble.

Each sugar will have characteristic crystal form of osazones.
 HC=NNHC6H5
C=NNHC6H5
OHCH
HCOH
HCOH
CH2OH
Glucososazone; Needle shaped crystals arranged like a broom or
sheaves of corn
Differences in glucose, fructose and mannose are dependent on the
first and second carbon atoms, and when the osazone is formed these
differences are masked.
Lactose is the sugar present in milk.
It is a reducing disaccharide.

Lactose forms osazone which

resembles "pincushion with
pins" or "hedgehog" or flower of
"touch-me-not" plant.
Maltose is a reducing disaccharide.

Sunflower shaped or petal

shaped crystals of Maltosazone
ACTION OF STRONG ALKALI ON SUGARS
• When treated with strong alkali, sugars are enolised and the
double bond will shift from 1:2 to 2:3, 3:4 or 4:5 to form the
corresponding ene diols and gets fragmented.
• The fragmented molecules will polymerise to form a yellow
resinous material called Caramel.
ACTION OF ACID ON SUGARS

• Dilute acids –
Aldoses in general are relatively stable.
Ketoses degraded.

• Concentrated acids –
Dehydrated to cyclical derivatives
Pentoses: Furfural
Hexoses : Hydroxy methyl furfural
• Ketoses are more susceptible to the action of acids
Action of Acids on Sugars

HC CH HC CH

HC C-CHO C C-CHO
HOCH2
O O

Furfural Hydroxymethyl Furfural

 Oxidation

1
2
3
4
5
6

Gluconic acid
 1 1
2 2
3 3
4 4
5 5
6 6

Gluconic acid
Glucuronic Acid Used for conjugation
with insoluble molecules
 1 1
2 2
3 3
4 4
5 5
6 6

Gluconic acid Glucuronic Acid Gluco-saccharic

acid
 Reduction

Glucose Sorbitol Fructose Mannitol

 Glucose is reduced to sorbitol
Mannose to mannitol
Fructose to sorbitol and mannitol.
Galactose is reduced to dulcitol

Sorbitol, mannitol and dulcitol are used to identify bacterial

colonies.
Mannitol is also used to reduce intracranial tension by forced
diuresis.
The osmotic effect of sorbitol and dulcitol produces changes in
tissues when they accumulate in abnormal amounts, e.g. cataract of
lens.
Formation of esters
GLYCOSIDES
Hemi-acetal group is condensed with
an alcohol or phenol group, it is called a glycoside

The non-carbohydrate group is called aglycone.

Glycosides do not reduce Benedict's reagent, because the sugar group is masked.
GLYCOSIDES
AMINOSUGARS
Glucosamine is seen in hyaluronic acid, heparin and blood
group substances.
Galactosamine is present in chondroitin of cartilage, bone and
tendons.
N-acetyl-glucosamine (GluNac)is seen in glycoproteins,
mucopolysaccharides and cell membrane antigens.

Amino sugars will not show reducing property. Will not produce
osazones.
L- FUCOSE/ 6-DEOXY-L-GALACTOSE

• Occurs in glycoproteins and in blood group substances

PENTOSE SUGARS OF NUCLEIC ACIDS
COMMON SUGAR SUBSTITUTE

Acesulfame-Potassium (Ace K) is made from acetoacetic acid.

It is 200 times sweeter than sugar; but calorie content is negligible.

Aspartame is made from aspartic acid and phenyl alanine, both are amino
acids. It is 200 times sweeter than sugar. Aspartame is not suitable for people
with phenyl ketonuria.

Sucralose is made from sucrose. Sucralose is approximately 600 times as sweet

as sugar.

Saccharin (Benzoic sulfimide) is an artificial sweetener. Although saccharin has

no food energy, it can trigger the release of insulin in humans and rats, apparently
as a result of its taste.

Cyclamate (Sodium cyclohexyl sulfamate) is 30–50 times sweeter than sugar.

 Dietary Fiber

Dietary fiber
Dietary fibre is contributed by the unavailable carbohydrates
in the diet. They contribute the bulk and assist in normal
bowel movements.
Cellulose, hemicellulose, pectin, alginates, and gums are the usual
glycans which form dietary fibre.
Cellulose is found in bran, flour, and tubers. Pectins are found in fruits.
Gums and alginates are found in legumes and oatmeal.
 Aquasomes

AQUASOMES
They are peptide/protein carriers with three layered self- assembled structures.
They comprise the central solid nanocrystalline core coated with
polyhydroxy oligomers onto which biochemically active molecules are
adsorbed.
The solid core provides structural stability. The carbohydrate coating acts as a
protectant and stabilizes the biochemically active molecules.
As the conformational integrity of bioactive molecule is maintained,
aquasomes are being proposed as a carrier system for the delivery of peptide
based pharmaceuticals. The system has been successfully utilized for the
delivery of insulin, hemoglobin and various antigens. Oral delivery of
enzymes like serratiopeptidase has also been achieved.
Thank you