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
• Monosaccharide
1

• Oligosaccharide
2

• Polysaccharide
3
 Monosaccharide
Cannot further Hydrolyzed
Oligosaccharide
• Oligosaccharides(Greek: oligo-few) contain 2-1O
monosaccharide molecules
• Joined by glycosidic bond
Polysaccharides
Containmore than 10
monosaccharide units.
 Polysaccharides

Homopolysaccharides Heteropolysacchrides

Starch
Glycogen
Agar
Mucopolysaccharide
Cellulose
Inulin
Dextrans
Chitin
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
carbon atoms by the formula 2n 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 Fischer's Haworth’s
formula formula
projection
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 12–18 hours it changes
to +52.5o.

This change in rotation with

time is called mutarotation.
Mutarotation

ɑ-D-glucopyranose ɓ-D-glucopyranose
Rotation of 112O Rotation of 190

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.

Lactose

It is the sugar present in milk

Lactose intolerance
Maltose
Isomaltose
REACTIONS OF
CARBOHYDRATE
Benedict’s 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
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?
Molisch’s 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
Fehling’s test

Same principle as benedicts test

Fehling’s A contains 7% copper
sulphate and Fehling’s B contains
sodium potassium tartarate.
Barfoed’s test
This test is based on the same principle
as Benedict’s test.
But, the test medium is acidic.
In acidic medium (pH 4.6)
monosaccharides react faster
than disaccharide.
Barfoed’s reagent contains copper
acetate
in glacial acetic acid.
Scanty Red precipitate at
bottom of tube
Seliwanoff’s test
Seliwanoff’s test is a chemical test
which distinguishes between aldose
and ketose sugars.
Ketohexoses like fructose on
treatment with HCl form 5-
hydroxymethylfurfural, 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
Retinopathy
Nephropathy
Neuropathy
Lactulose

• Lactulose is also known as beta-D-

galactopyranosyl-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, O-
linked.
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
Esterificationof 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 constituents of
(GluNac) and N- glycoproteins,
acetyl galactosamine Mucopolysaccharide and cell
(GalNac) 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
Glycoge
n
Cellulos
e
Inulin
Dextrans
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.
 Short
Hydrolysis of starch time to
long time
Amylodextrin = 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
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 10–15 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 . Fibers

Decreases
serum
cholesterol
level 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 micro- organisms.
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 N-
acetylglucosamine with beta-1,4
glycosidic linkages.
Heteropolysaccharides

• Agar
• Mucopolysaccharide
 Agar

Agar = The linear polysaccharide

Agarose
+ agaropectin
It is dissolved in water at 100ºC, 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
e. glycosaminoglycans(GAGs)
[acidic sugar–amino 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.

Thisproduces 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 D-glucuronic acid Connective tissue lubricant and shock

acid and N-acetyl D- Synovial fluid absorbant in joints
glucosamine Vitreous humor
Gel around ovum

Chondroitin D-glucuronic acid and N- bone, cartilage, Helps to maintain

sulphate acetyl D-galactosamine Tendons,heart the structure And
4-sulfate. valves and skin. shapes of tissues

Dermatan D-Iduronic acid and Skin Helps to maintain

sulfate N-acetyl D-galactosamine shapes of tissues
4 –sulfate.
Keratan galactose and N-acetyl cornea Keeps cornea
sulphate glucosamine tendons Transparent

Heparin sulphated glucosamine blood, lung, liver Anticoagulant

and glucuronic acid or ,kidney, spleen Clearing factor
iduronic acid
Hyaluronic acid

N-Acetyl-glucosamine → beta-1, 4-
Glucuronic 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
 DDeerrmmaattaa
nn

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

Heparan
Lipoprotein HS
EN
sulphate
lipase

Heparin displaces
On capillary
lipoprotein lipase from
endothelial
heparan sulphate binding
wall surface
site hence clearing factor
 Proteoglycans and
Glycoproteins

Proteoglycans: When carbohydrate

chains are attached to a polypeptide
chain.

Glycoproteins: Carbohydrate content ≤

10%.
 Proteoglycans

Figure showing Proteoglycans aggregate

Glycoprotein Major function
Glycophorin glycoprotein of
erythrocytes cell
membrane
Collagen Structure of cartilage
and bone
Ceruloplasmin Transport protein
Immunoglobulin Defense against
infection
Intrinsic factor Absorption of vitamin
B12
Fibrinogen Blood clotting
 Thank
you

Any questions?