Carbohydrate chemistry

Dr. Herat D. Soni

Assistant professor
Rural medical college
Loni

Definition
Carbohydrates

may be defined as
polyhydroxy aldehydes or
ketones or compounds which
produce them on hydrolysis.
Formula = (C.H2O)n

Biomedical Importance

Most abundant dietary source of energy. Brain cells

and RBCs are almost wholly dependent on
carbohydrates as the energy source.
Also serve as storage form of energy Glycogen.
Carbohydrates are precursors for many organic
compounds (fats, amino acids).
Participate in the structure of cell membrane &
cellular functions (cell growth, adhesion and
fertilization).
Certain carbohydrate derivatives are used as
drugs, like cardiac glycosides / antibiotics.
DM (diabetes mellitus)

Sources

CLASSIFICATION OF
CARBOHYDRATE

Classification
1

Monosaccharide
Oligosaccharide

Polysaccharide

Monosaccharide
Cannot further Hydrolyzed

Oligosaccharide

Oligosaccharides(Greek: oligo-few) contain 2-1O

monosaccharide molecules
Joined by glycosidic bond

Polysaccharides

Contain more than 10 monosaccharide

units.

Polysaccharides

Homopolysaccharides

Starch
Glycogen
Cellulose
Inulin
Dextrans
Chitin

Heteropolysacchrides

Agar
Mucopolysaccharide

Isomers
Same chemical formula but different
structural formula
Example = Glucose and fructose

C6H12O6

Stereoisomer

Same chemical and structural formula but

differ in spatial configuration.

Asymmetric carbon atom

Asymmetric carbon means that four
different groups are attached to the same
carbon.
The reference molecule is glyceraldehyde
which has a single asymmetric carbon
atom.
The number of possible stereoisomer
depends on the number of asymmetric
n
carbon atoms by the formula 2 where n is
the number of asymmetric carbon atoms.

Reference Carbon Atom of

Sugars
All monosaccharide can be considered as molecules
derived from glyceraldehyde by successive addition of
carbon atoms. Therefore, penultimate carbon atom is the
reference carbon atom for naming the mirror images

D and L isomerism(Enantiomers)

D-sugars are naturally occurring sugars and

body can metabolize only D-sugars.
D-glucose is dextrorotatory. In clinical practice,
it is often called as dextrose

Optical isomerism(d and l)

The presence of asymmetrical carbon atom

causes optical activity. When a beam of
plane-polarized light is passed through a
solution of carbohydrates, it will rotate the
light either to right or to left.
Right = dextrorotatory (+) (d)
Left = levorotatory (-) (l)
D-glucose is dextrorotatory but D-fructose
is levorotatory
Equimolecular mixture of optical isomers
has no net rotation (racemic mixture)

Epimers

When sugars are different from one

another, only in configuration with
regard to a single carbon atom,
other than the reference carbon
atom, they are called Epimers.

Epimers

Anomers
To understand this we first understand
Three Representations of Glucose
Structure

open chain
projection
formula

Fischer's
formula

Haworths
formula

The 1st carbon, aldehyde group is

condensed with the hydroxyl group of the
5th carbon to form a ring. Ring structure
represents hemi acetal form.
Glucose exists in biological systems not
as a rectangle, but as a pyranose ring.
b-D-glucopyranose is the predominant
form (63%).

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.
These carbon atoms are known as
anomeric carbon atoms.

Fischer's formula

Haworth formula

Anomeric carbon atom

Mutarotation
When

D glucose is crystallized at
room temperature, and a fresh
solution is prepared, its specific
rotation of polarized light is +112o;
but after 1218 hours it changes
o
to +52.5 .

This

change in rotation with time

is called mutarotation.

Mutarotation
-D-glucopyranose
O
Rotation of 112

-D-glucopyranose
0
Rotation of 19

1/3 are alpha type and 2/3rd are beta

variety to get the specific rotation of +52.5o

DISACCHARIDES
Sucrose
Maltose
Isomaltose
Lactose.

Sucrose

It is the sweetening agent known as cane

sugar.
It is present in sugarcane and various fruits.

Hydrolysis of sucrose (optical rotation

+66.5) will produce one molecule of
glucose (+52.5) and one molecule of
fructose (92).
Therefore, the products will change the
dextrorotation to levorotation, or the plane
of rotation is inverted.
Equimolecular mixture of glucose and
fructose thus formed is called invert sugar.
The enzyme producing hydrolysis of sucrose
is called sucrase or invertase.
Honey contains invert sugar.
Invert sugar is sweeter than sucrose.

Lactose

It is the sugar present in milk

Lactose intolerance

Maltose

Isomaltose

REACTIONS OF
CARBOHYDRATE

Benedicts test

Principle
The principle of Benedict's test is that
when reducing sugars are heated in the
presence of an alkali(pH 10.6), they get
converted to powerful reducing
compounds known as enediols.
Enediols reduce the cupric ions (Cu2+)
present in the Benedict's reagent to
cuprous ions (Cu+) which get precipitated
as insoluble red copper oxide.

Detect the presence of glucose in urine

(glucosuria).
It is a standard laboratory test employed
for follow-up of diabetes mellitus in PHC.
Benedict's reagent contains sodium
carbonate, copper sulfate and sodium
citrate
Any sugar with free aldehyde/keto group
will reduce the Benedict's reagent.
Therefore, this is not specific for glucose.

Carbohydrates giving positive

Benedict s test:
Glucose, Fructose, Galactose
Lactose, Maltose
Sucrose ???????
Starches do not react or react very poorly
with Benedict's reagent, due to the
relatively small number of reducing sugar
moieties, which occur only at the ends of
carbohydrate chains.

Non-Carbohydrates giving
positive Benedict s test
High concentration of Uric acid and
Ketones
Homogentisic acid (solution turns black
due to black colored oxidized
homogentisic acid)
Vitamin C (even without Boiling)
Certain drugs like aspirin, cephalosporins

Glucose oxidase test

Glucose + O2
Glucose Oxidase

Gluconolactone + H2O2
H2O2 + (reduced colourless dye)
Peroxidase
(Oxidized colored dye)

Reagents for this test are present on a strip

of paper in solid form.
When the paper is wet with urine, the
reagents dissolve in urine on paper and
react with glucose in urine.
The darkness of color can be correlated
with amount of glucose present in urine.
Because Glucose oxidase enzyme can act
only on beta-D Glucose, other reducing
substances do not give this test positive.

 Thus,

compounds like Vitamin C,

Aspirin utilize H2O2 produced in the
reaction.
Due to lack of H2O2, Peroxidase can
not oxidize dye. Thus, glucose may
not be detected even if present, if
urine contain Vitamin C or Aspirin in
large amount. This phenomenon is
called false negative result.

Osazone Formation
All

reducing sugars will form

osazones with excess of phenyl
hydrazine when kept at boiling
temperature.
Glucose, Galactose and Fructose will
produce the same needle-shaped
crystals. Why?

Molischs test
All carbohydrates when treated with conc.
sulphuric acid undergo dehydration to
give fufural compounds. These
compounds condense with Alpha-napthol
to form colored compounds.
Molish test is given by sugars with at
least five carbons because it involves
furfural derivatives, which are five carbon
compounds.

Purple ring at junction

Fehlings test
Same

principle as benedicts test

Fehlings A contains 7% copper
sulphate and Fehlings B contains
sodium potassium tartarate.

Barfoeds test
This test is based on the same principle as
Benedicts test.
But, the test medium is acidic.
In acidic medium (pH 4.6)
monosaccharides react faster than
disaccharide.
Barfoeds reagent contains copper acetate
in glacial acetic acid.

Scanty Red precipitate at

bottom of tube

Seliwanoffs test
Seliwanoffs

test is a chemical test

which distinguishes between aldose
and ketose sugars.
Ketohexoses like fructose on
treatment with HCl form 5hydroxymethylfurfural, which on
condensation with resorcinol gives a
cherry red complex.

Cherry
red color

Oxidation

The glucuronic acid is used by the body for

conjugation with insoluble molecules to make them
soluble in water for detoxification purpose and also
for synthesis of heteropolysaccharides.

Reduction to Form Alcohols

When treated with reducing agents
hydrogen can reduce sugars. Aldose yields
corresponding alcohol.
Glucose is reduced to sorbitol
mannose to mannitol
fructose becomes sorbitol and mannitol
Galactose is reduced to dulcitol and
ribose to ribitol.

Significance of reduction
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 produces
changes in tissues when they
accumulate in abnormal amounts, e.g.
cataract of lens.

Cataract
Retinopathy
Nephropathy
Neuropathy

Lactulose

Lactulose is also known as beta-Dgalactopyranosyl-D-fructofuranose.

Used in constipation

Glycosides

The hydroxyl group of anomeric carbon of a

carbohydrate can join with a hydroxyl group
of another carbohydrate or some other
compound to form a glycoside and the bond
so formed is known as glycosidic bond.
eg. R-OH + HO-R
R-O-R' + H2O
The non-carbohydrate moiety is known as
aglycone phenol, sterol, glycerol and
methanol.
Glycosidic bond can be N-linked or, Olinked.

Biomedical importance of
glycosides

Cardiac Glycosides Digoxin, Digitoxin

Used in cardiac insufficiency.
Stimulate cardiac muscle contraction.
Contain steroids as aglycone
component.

Ouabain Sodium pump inhibitor.

Streptomycin
Antibiotic
Given in Tuberculosis

Phloridzin
cause renal damage, glycosuria.
Blocks the transport of sugar across the
mucosal cells of small intestine & also
renal tubular epithelium.

Formation of Esters
Esterification

of alcoholic groups of
monosaccharides with phosphoric
acid is a common reaction in
metabolism.
Examples :Glucose-6-phosphate, and
Glucose-1-phosphate.
ATP donates the phosphate moiety.

Amino sugars

Amino groups may be substituted for hydroxyl

groups of sugars to give rise to amino sugars

Importance
Amino sugars

Found in

Glucosamine

Hyaluronic acid, heparin and

blood group substances

Galactosamine

Chondroitin sulphate of
cartilage, bone and tendons.

Mannosamine

constituent of glycoproteins

N-acetylglucosamine
(GluNac) and Nacetyl galactosamine
(GalNac)

constituents of
glycoproteins,
Mucopolysaccharide and cell
membrane antigens.

Deoxy Sugars
Oxygen of the hydroxyl group may be removed to
form Deoxy sugars

2-deoxy D-ribose is important

part in DNA.

Homopolysaccharides
Starch
Glycogen
Cellulose
Inulin
Dextrans
Chitin

Starch
It

is the reserve carbohydrate of plant

kingdom
Sources: Potatoes, cereals (rice,
wheat) and other food grains.
Starch is composed of amylose and
amylopectin.

 Amylose

is made up of glucose units

with alpha-1,4 glycosidic linkages to
form an unbranched long chain. Water
soluble.
The insoluble part absorbs water and
forms paste like gel; this is called
amylopectin.
Amylopectin is also made up of
glucose units, but is highly branched.
The branching points are made by
alpha-1,6 linkage

Iodine test for starch

Starch

will form a blue colored

complex with iodine; this color
disappears on heating and reappears
when cooled. This is a sensitive test
for starch.

Starch

is nonreducing because the

free sugar groups are negligible in
number.

Hydrolysis of starch
Amylodextrin

Short
time to
long time

= violet color with

iodine and is non-reducing.
Erythrodextrin = red color with
iodine and mildly reduce the
Benedict's solution.
Achrodextrins = no color with
iodine, reducing)
Maltose = (no color with iodine, but
powerfully reducing)

Glycogen

It is the reserve carbohydrate in animals.

It is stored in liver and muscle.
Liver glycogen stores increase during the
well-fed state , and are depleted during a
fast.
Glycogen is composed of glucose units
joined by alpha-1,4 links in straight
chains. It also has alpha-1,6 glycosidic
linkages at the branching points.
Glycogen is more branched and more
compact than amylopectin.

Liver and muscle glycogen

Cellulose
It is made up of glucose units combined
with beta-1,4 linkages.
It has a straight line structure, with no
branching points.
Beta-1,4 bridges are hydrolyzed by the
enzyme cellobiase. But this enzyme is
absent in animal and human digestive
system, and hence cellulose cannot be
digested.

Importance
It is a major constituent of fiber, the
nondigestable carbohydrate.
Fiber can absorb 1015 times its own
weight in water, drawing fluid into
the lumen of the intestine
Increasing bowel motility
1.Decrease the risk for constipation

2. Lower LDL cholesterol levels

Cholesterol

Intestine
Bile salt

Decreases
serum
cholesterol
level

Fibers

Excreted

Can bind various toxic substances

including carcinogens & eliminate them in
faecal matter
3.Decreases chances of some cancers
Delays gastric emptying and can result in a
sensation of fullness
4. Reduced peaks of blood glucose
following a meal

Inulin
It is a long chain homoglycan composed
of D-fructose units with repeating beta-1,2
linkages.
It is the reserve carbohydrate present in
various bulbs and tubers, such as onion,
garlic.
It is clinically used to find renal
clearance value and glomerular
filtration rate.

Dextrans
These are highly branched homopolymers
of glucose units with 1-6, 1-4 and 1-3
linkages. They are produced by microorganisms.
Since they will not easily go out of
vascular compartment, they are used for
intravenous infusion as plasma volume
expander for treatment of hypovolemic
shock.

Dextrose, Dextrin and Dextran

are different
D-glucose is otherwise called Dextrose, a
term often used in bed-side medicine, e.g.
dextrose drip.
Dextrin is the partially digested product of
starch.
Dextran is high molecular weight
carbohydrate, synthesized by bacteria.

Chitin
It

is present in exoskeletons of
insects.
It is composed of units of Nacetylglucosamine with beta-1,4
glycosidic linkages.

Heteropolysaccharides

Agar
Mucopolysaccharide

Agar
Agar = The linear polysaccharide Agarose
+ agaropectin
It is dissolved in water at 100C, which
upon cooling sets into a gel. Agarose is
used as matrix for electrophoresis.
Agar cannot be digested by bacteria and
hence used widely as a supporting agent to
culture bacterial colonies.

Mucopolysaccharide
i.e. glycosaminoglycans(GAGs)

[acidic sugaramino sugar]n.

Because of their large number of negative

charges, these heteropolysaccharide chains
tend to be extended in solution. They repel
each other, and are surrounded by a shell
of water molecules. When brought
together, they slip past each other.

This produces the slippery consistency

of mucous secretions and synovial fluid.

This property contributes to

the resilience of synovial fluid
and the vitreous humor of the
eye

GAGs

Composition

Tissue distribution Functions

Hyaluronic
acid

D-glucuronic acid
and N-acetyl Dglucosamine

Connective tissue
Synovial fluid
Vitreous humor
Gel around ovum

lubricant and shock

absorbant in joints

bone, cartilage,
Tendons,heart
valves and skin.

Helps to maintain
the structure And
shapes of tissues

Chondroitin D-glucuronic acid and Nsulphate

acetyl D-galactosamine
4-sulfate.
Dermatan
sulfate

D-Iduronic acid and

Skin
N-acetyl D-galactosamine
4 sulfate.

Helps to maintain
shapes of tissues

Keratan
sulphate

galactose and N-acetyl

glucosamine

cornea
tendons

Keeps cornea
Transparent

Heparin

sulphated glucosamine
and glucuronic acid or
iduronic acid

blood, lung, liver

,kidney, spleen

Anticoagulant
Clearing factor

Hyaluronic acid

N-Acetyl-glucosamine beta-1, 4Glucuronic acid beta-1-3-N-Acetyl

glucosamine and so on.

Hyaluronidase
Breaks b(1-4 linkages) in hyaluronic acid.
Present in high concentration in testes,
seminal fluid, and in certain snake and insect
venoms.
Hyaluronidase of semen clears the gel
(hyaluronic acid) around the ovum allowing a
better penetration of sperm into the ovum.
Serves important role in fertilization
Hyaluronidase of bacteria helps their invasion
into the animal tissues.

Chondroitin sulphate

glucuronic acid beta-1,3-N-acetyl

galactosamine sulfate beta-1, 4
and so on

Dermatan sulfate

L-iduronic acid and N-acetylgalactosamine

in beta-1, 3 linkages

Keratan sulphate

Only GAG not having Uronic acid.

Heparin

It contains repeating units of sulphated

glucosamine alpha-1, 4-L-iduronic acid
or glucuronic acid and so on

Heparin is an anticoagulant( prevents

blood clotting).

Heparin helps in the release of the enzyme

lipoprotein lipase which helps in clearing
the turbidity of lipemic plasma.
Lipoprotein lipase breaks TG in glycerol
and FFA.

Capillary

Lipoprotein
lipase

EN

HS

Heparin displaces
lipoprotein lipase from
heparan sulphate binding
site hence clearing factor

Heparan
sulphate
On capillary
endothelial
wall surface

Proteoglycans and
Glycoproteins

Proteoglycans: When carbohydrate

chains are attached to a polypeptide chain.

Glycoproteins: Carbohydrate content

10%.

Proteoglycans

Figure showing Proteoglycans aggregate

Glycoprotein
Glycophorin

Collagen
Ceruloplasmin
Immunoglobulin
Intrinsic factor
Fibrinogen

Major function
glycoprotein of
erythrocytes cell
membrane
Structure of cartilage
and bone
Transport protein
Defense against
infection
Absorption of vitamin
B12
Blood clotting

Thank
you
Any questions?