Carbohydrates
(Monosaccharides and simple sugars)
GENERAL STATEMENTS ABOUT CARBOHYDRATES:
 The most abundant group of organic molecules in nature.
 More than 75 % of the dry weight of plants.
 Less than 1 % of the weight of animals is carbohydrate.
 About 65 % of our diet is carbohydrate. In developing
countries the figure can be up to 80%
OVERVIEW
 The term carbohydrate originated from the empirical
formulas of several compounds in this class.
 Glucose (C6H12O6) can be represented as a “hydrate of
carbon” C6(H2O)6
 Now carbohydrates are described as polyhydroxy aldehydes
and ketones or derivatives of such compounds
 The chemistry of carbohydrates is mainly the chemistry of two
functional groups; the hydroxyl and the carbonyl groups
 Carbohydrate chemistry is also the chemistry of acetals and
hemiacetals, ketals and hemiketals
 Can be formed by the interaction of the hydroxyl and
carbonyl groups
The main functions of carbohydrates are:
1. To serve as energy sources (also storage form of energy)
2. To serve as structural component of many organisms; cell
walls of bacteria, exoskeleton of many insects, fibrous
cellulose of plants
3. They can also serve as building blocks in the synthesis of
other biomolecules (for example the nucleic acids DNA
and RNA)
 Most Carbohydrates have the Empirical formula
Cn (H2O)n [or Cn H2n On]
 One molecule of water for each carbon (hydrates of
carbon) (CH20)n
 Most simple sugars contain a single carbon chain with
multiple OH groups and contain either Aldehyde or Ketone
groups
 If the sugar contains an aldehyde it is called an Aldose
 If the sugar contains a ketone it is called a Ketose
Chirality:
All carbohydrates are chiral molecules (exist in a number of
different orientations)
 Usually classified as D and L according to the
arrangement of the OH group on the carbon furthest from
the carbonyl group.
 System is based on a comparison with the stereochemistry
of the simplest carbohydrate glyceraldehyde (C3H6O3)
 Most carbohydrates in nature have the D configuration.
(Note: How about amino acids?)
 CHO CHO
H OH HO H
CH2OH CH2OH

D-Glyceraldehyde L-Glyceraldehyde
 The
D and L forms of any specific compound are Enantiomers (mirror
images) of each other.
The carbonyl group of carbohydrates is very reactive and can Carbohydrates can exist as either linear (open chain) or
react with as cyclic structures. For C5, C6 and C7 carbohydrates
 water to form Gem-diols the cyclic form is the most common.

 OH groups to form Hemi-acetals, acetals] [Hemi-ketals

and ketals]

 In solution the Carbonyl groups of Aldoses and Ketoses react

with Water to form Gem-Diols
 2 OH groups bound to the same (geminal) carbon
 Gem-diol formation is freely reversible, although it can be a
• Fischer projection formula: ACYCLIC
relatively SLOW process (seconds to minutes), depending on
• Emil Fischer (1852-1919): pioneering work on
the structure of the SUGAR. carbohydrates, proteins and dyestuffs
 Nobel prize winner in 1902
• Haworth Projection formula; CYCLIC
HEMIACETALS, HEMIKETALS, ACETALS, KETALS
• W.N. Haworth British carbohydrate chemist who won
 Aldehyde + alcohol  hemiacetal the Nobel prize in 1937
 Ketone + alcohol  hemiketal
CYCLIC FORMS OF MONOSACCHARIDES
Because carbohydrates contain OH and C=O groups, they can
react to form hemiacetals or hemiketals. • > 6 membered rings with 5 carbons + 1 oxygen are
called pyranose rings
 Hemiacetals can react with alcohols to form Acetals
• > 5 membered rings with 4 carbons + 1 oxygen are
 HemiKetals can react with alcohols to form Ketals
 Ketoses react with Alcohols to form HemiKetals or Ketals called furanose rings
 HemiAcetals and HemiKetals formation is reversible • Pyranose rings are more common
 Acetals and Ketals are stable (i.e. not broken down
except under extreme conditions)
 In solution an equilibrium exists between the open chain ANOMERS
and the two cyclic forms of a carbohydrate. This
 For 5, 6 or 7 carbon carbohydrates, after cyclisation the
equilibrium is called mutarotation.
carbonyl carbon becomes a chiral center.
Two different orientations are possible about this carbon.

These are called a and b anomers and the carbon is

called the anomeric carbon.

[The oxygen from this OH group corresponds to the

carbonyl oxygen in the Fischer projection]

 If the OH group at the anomeric carbon is on the same

side of the ring as the CH2OH group
 (i.e. both above or both below the plane of the ring)
it is called the b anomer.

 If the OH group at the anomeric carbon and the CH2OH

EPIMERS groups are on opposite sides of the plane of the ring
 Carbohydrates vary by the number and position of the (one above the ring and one below the ring)it is called
OH groups the a anomer.
6
 Two carbohydrates that differ only by the orientation of CH2OH
5
O
a single OH group are called epimers. H H
4
H 1
a D-glucose and
OH H
O H OH 3 OH b D-glucose are
1 2
ANOMERS
2 OH H OH
HO 3  D-Glucose
6
4 OH CH2OH
5 OH H
5
O OH
6 OH 4
H 1
OH H
OH 3 H
2

H OH
 D-Glucose
FIVE (5) CARBON SUGARS OTHER MONOSACCHARIDES

 The most common 5 carbon sugar is D-Ribose

 It is a building block of a number of biomolecules

including DNA and RNA (deoxyribonucleic acid and
ribonucleic acid)and also molecules involved in energy
transfer such as ATP (Adenosine Triphosphate)

 Another 5 carbon sugar found in the body is Arabinose.

It is a building block of several Anticancer drugs

SIX (6) CARBON SUGARS

N-acetylneuraminic acid (Sialic Acid)
 D-GLUCOSE (also known as Dextrose or Blood sugar) is
the primary source of Carbon atoms and also Energy for
• Carbohydrates with 7 or more carbons are rare
most tissues in the body.
• The 9 carbon sugar Neuraminic acid and its N-acetyl
derivative Sialic Acid is found in nerve cells
 It is also the only nutrient for the BRAIN

REACTIONS OF SIMPLE CARBOHYDRATES

 Galactose, Mannose and Fructose are other important 6
Carbon sugars. They can also be converted (via
phosphate derivatives) to Glucose • Mutarotation
• Phosphate formation
Galactosemia is a genetic disease in which the enzyme • Oxidation
responsible for Galactose breakdown is missing.
• Reduction
Accumulation of Galactose leads to damage to the liver
• Amine formation
and brain. The most effective treatment is early diagnosis
and removal of Galactose from the diet • Polymerization
• Reducing sugars
Mutarotation • Oxidation of both terminal carbon atoms to carboxyl
groups produces an aldaric acid
• Conversion between linear and cyclic forms the cyclic
(Haworth) form is a result of hemiacetal or hemiketal
formation.
Phosphate formation

• OH groups of carbohydrates (like other OH groups) can

react with phosphoric acids to form phosphate esters.
R-OH  R-O-PO3 2-

Oxidation

• If the aldehyde group of Carbohydrates is oxidised

• ( CHO  COOH ) the product is called an Aldonic Acid
• For Glucose the product is called Gluconic Acid
• For ribose the product is Ribonic Acid
• If the terminal OH group of Carbohydrates is oxidised
( CH2OH  COOH )
• The product is called a Uronic Acid (sugar acids)
• For Glucose the product is called Glucuronic Acid
(Vitamin C is a derivative of Glucuronic Acid
Reduction
• The aldehyde or ketone group of carbohydrates can be
reduced to form sugar alcohols (also called alditols)
R-CHO  R-CH2OH
• Alditols are used in processing foods and
pharmaceuticals
• The Alditol of Glucose is called Sorbitol (or glucitol)
• It is sometimes given to diabetics.
• It is approximately 60% as sweet as glucose

Amino Sugars

• An OH group of carbohydrates is often replaced by an

amino group NH2
• The Amino group is sometimes acetylated
(NH2  NHCO-CH3)
• Glucosamine and N-Acetyl Glucosamine are amino Reagents: Fehling’s and Benedict’s reagents
derivatives of Glucose.
• They are important components of structural
carbohydrates and are often linked to other
biomolecules (proteins or lipids)

Negative: Still blue

Positive: Brick Red Precipitate

Polymerization (Carbohydrates can link together)

• 2 units linked together is called a Disaccharide (dimer)

• 3 units is called a Trisaccharide (trimer)
• Several units is called a Oligosaccharide (oligomer)
• Many units is called a Polysaccharide (polymer)

Reducing and non-reducing sugars

• Saccharides that have a hemiacetal or hemiketal

structure are in equilibrium with an acyclic
hydroxyaldehyde or hydroxyketone form
• Readily reduce various metallic ions
Carbohydrates NOMENCLATURE:
(Polysaccharides) Most mono- and disaccharide names end in - ose

 Mono and disaccharides taste sweet and are

sometimes called sugars.
 They are all water soluble.

 Monosaccharides 1 carbohydrate unit

 Disaccharides 2 carbohydrate units
 Polysaccharides > 2 carbohydrate units (up to
1,000,000)

Disaccharides and polysaccharides are formed by

glycosidic bonds

The anomeric hydroxyl group and a hydroxyl group of

another sugar or some other compound can join together,
splitting out water to form a glycosidic bond.
R-OH + HO-R' --> R-O-R' + H2O
Carbohydrate units can link together
Maltose is a disaccharide with an a (1,4) glycosidic linkage
Carbohydrate units can also link together to form polymers. between the C1 hydroxyl of one glucose and the C4
Monosaccharides - 1 sugar unit hydroxyl of a second glucose. Maltose is the  anomer,
because the O at C1 points down from the ring.
examples: ribose (5 C), glucose, fructose, galactose (6 C)
Disaccharides - 2 sugar units
example: sucrose, maltose and lactose
Polysaccharides – more than two sugar units
example: amylose, starch, cellulose
Oligosaccharides: few sugars linked together (less than 10)
Cellobiose, a product of cellulose breakdown, is the SUCROSE a(1,2) glycosidic bond
otherwise equivalent  anomer. 6 1
1H OH
CH
6 OH C 2 OH
CH2
2OH CH2
• The  (1,4) glycosidic linkage is represented as a "zig- H
5
5 O
O HH O
H
H
zag" line, but one glucose residue is actually flipped
H
H 2 O
4HH 1 + 2 H HO 5
4O H 1 + H HO 5
over relative to the other. OH O3
H H OH HO 3 CH2OH
OH 3 2 OH HO 3 4 6CH OH
2 4 6 2
H OH OH H
H OH OH H
 D-Glucose  D-Fructose
 D-G lucose  D-Fructose
6 1
CH OH C
6 2 1H2OH
C5 H2OH
O CH2OH
5
O O
HO
4 1 2 5
OH O
4 1 2
OH O3H
O HO CH5 OH
6 2
2 3 4
OH 3 O
O2H 3 4
CH2OH
OH 6
OH OH
Other disaccharides: LACTOSE b (1,4) glycosidic bond

• Sucrose, common table sugar, has a glycosidic bond 6

CH2OH 6
CH2OH
linking the anomeric hydroxyls of glucose and 5
O 5
O
OH
fructose. Because the configuration at the anomeric H
4 1 +
carbon of glucose is  (O points down from the ring), OH 4
OH
1

OH
the linkage is designated (1,2). The full name is -D-
3 OH
2
3 OH
2
glucopyranosyl-(1,2)-D- fructopyranose. OH OH
 D-Galactose  D-Glucose
• Lactose, milk sugar, is composed of glucose and
galactose with (1,4) linkage from the anomeric 6
CH2OH
hydroxyl of galactose. Its full name is -D- 5
O
galactopyranosyl-(1,4)--D-glucopyranose 6
CH2OH 4 1
OH
5
O O 3 OH
OH 2
4
OH 1 OH
3
2

OH
Lactose (milk sugar)
• Lactose consists of a Galactose and a Glucose linked by
a b-1,4 glycosidic bond
i.e. C1 of Galactose is bonded to C4 of Glucose
• The orientation is b for the anomeric bond of the
Galactose
(i.e. the bond is above the plane of the ring)
Lactose makes up about 40% of an infant’s diet (up to 1
year). Approximately 4-6 % of cow’s milk and 6-8 % of
human milk is Lactose.
• Some people have Lactose Intolerance, they are
unable to digest the Lactose in milk.
- caused by a deficiency of the enzyme Lactase
It is treated by
 avoiding Lactose in the diet or
 by giving the patient extra Lactase
 some special forms of milk have been pretreated with
Lactase (or are made lactose-free)
REDUCING AND NON REDUCING SUGARS
• In sucrose both of the anomeric bonds are joined
together,
i.e. C1 from Glucose and C2 from Fructose
• Mutarotation cannot occur and the sugar cannot be
converted to the open chain form (Fischer projection).
Sucrose is NOT a reducing sugar because the aldehyde or
ketone form of the sugars cannot exist
Lactose and Maltose are Reducing sugars because in each
case one of the anomeric carbons is available for
mutarotation.
• The open chain form contains an aldehyde group
Reducing sugars
• Oxygen of the anomeric carbon of a sugar is not
attached to any other structure
• Can react with chemical reagent’s (Benedict’s and
Fehling’s solutions) and reduce the active component –
we will be doing this in the laboratory
• The anomeric carbon itself becomes oxidized
Disaccharides can be hydrolysed to monosaccharides
• Sucrose  glucose + fructose
• Maltose  glucose + glucose
• Lactose  Galactose + glucose
• The human body is unable to use disaccharides or
polysaccharides directly.
• They must be broken down into monosaccharides.
• Hydrolysis reactions are always catalysed by enzymes
• Maltase breaks up the sugar Maltose into 2 Glucose units
• The enzyme Sucrase breaks up the sugar Sucrose into
Glucose and Fructose units
• The enzyme Lactase breaks up the sugar Lactose into
Glucose and Galactose units
Polysaccharides Glycogen

• are used as Storage forms of energy or as Structural • The main storage form of glucose in animals .
Materials
• It also contains a 1,4 and a 1,6 glycosidic bonds, but it is
• They have many carbohydrate units (up to 1,000,000) more branched than starch.
• They are all non-reducing carbohydrates
• It is broken down to glucose units by the enzyme a-
• They do not undergo mutarotation (ring opening)
glucosidase
• They are NOT sweet tasting
• They have limited solubility in water General cyclic structure of a 6 carbon carbohydrate
• The three most common polysaccharides found in (sugar) for simplicity, the extra OH groups are omitted
nature are Cellulose, starch and glycogen
• They all contain D-glucose subunits.
• They differ by the way the subunits are linked together
(a 1,4; a 1,6; b 1,4 etc).
• Glycogen, starch, cellulose can all be hydrolysed to
give thousands of glucose subunits
Glycogen (-1,4 and -1,6 linkage)

Starch
CH2OH CH2OH
H O O
glycogen
H H H
H H
OH H OH H 1
• The main sstorage form of glucose in plants OH
O
O
H OH H OH

There are two forms of Starch CH2OH CH2OH 6 CH2 CH2OH CH2OH
H O H H O H H 5 O H H O H H O H
H H H H H
• Amylose (20%), which is less branched and composed OH H
O
OH H
O
4 OH H 1
O
4 OH H
O
OH H
OH
OH
mainly of a-1,4 Glycosidic bonds 3 2
H OH H OH H OH H OH H OH

• Contains 250-4000 subunits arranged in a helix

• Amylopectin (80%), which is similar to glycogen and has Amylose (-1,4 linkage)
both a-1,4 and a-1,6 linkages. 6CH OH
CH2OH 2 CH2OH CH2OH CH2OH
O
Starch is found in many foods including rice, pasta,
O 5 O O O H H
• H
H
H H
H
H H
H
H H
H
H
H
OH H
OH H 1 4 OH H 1 OH H OH H
potatoes, wheat and cereals OH
O
3 2
O O O OH

H OH H OH H OH H OH H OH

• Starch is broken down to glucose units by a group of amylose

enzymes called Amylases
 Cellulose (-1,4 linkage)

CH2OH 6CH OH CH2OH CH2OH CH2OH

2
O 5 O O H O H O OH
H H H
H H H H H
OH H 1 O 4 OH H 1 O OH H O OH H O OH H
OH H H H
H 2 H
3
H OH H OH H OH H OH H OH
cellulose

HOMOPOLYMERS AND HETEROPOLYMERS

Homopolymers

Starch, glycogen and cellulose all contain the same type

Amylopectin (-1,4 and -1,6 linkage) of monosaccharide units. If they are completely hydrolysed
they will all give Glucose. (only 1 type of monosaccharide
CH2OH CH2OH
amylopectin subunits)
H O H H O H
H H
Heteropolymers
OH H OH H 1
O
OH
O
H OH H OH

CH2OH CH2OH 6 CH2 CH2OH CH2OH Contain different monosaccharide subunits (for example
H
H
O H H
H
O H H 5
H
O H H
H
O H H
H
O H
Glucose and Arabinose, Glucose and Galactose etc)
OH H OH H OH H 1 4 OH H OH H
4 O O
O O
Heteropolymers often contain amino or acetylated amino
OH OH
3 2
H OH H OH H OH H OH H OH
sugars such as Glucosamine, N-Acetyl Glucosamine or
Sialic Acid

Cellulose is a structural component of plant cells.

DIETARY FIBER
• It is used to construct rigid cell walls in plants.
• It is the most abundant organic substance on earth. • Is the general name given to materials from plants that
• Cellulose contains b- 1,4 glycosidic linkages the body cannot digest
• Humans and other animals have enzymes that can • Fiber helps digestion by improving bowel function.
break down starch or glycogen but not cellulose. • Soluble fibers consist of molecules called Pectins & Gums
• Bacteria have enzymes that can break down cellulose. – polysaccharides with chemically modified sugar units.
• Some animals such as cows, have bacteria in their • The main insoluble fiber is Cellulose
stomachthat break down cellulose. • Increased levels of fiber in the diet lead to reductions or
• It is broken down by the enzyme Cellulase improvements in a variety of conditions including heart
disease, colon cancer and diabetes
Other polysaccharides CHITIN
Pectins is a soluble polysaccharides often found in fruits • This is a polymer of Glucosamine.

Pectin is a complex polysaccharide consisting mainly of • It forms a very rigid polymer.

esterified D-galacturonic acid resides in an alpha-(1-4)
• Used by many insects and other animals to form tough
chain. The acid groups along the chain are largely esterifed
exoskeleton
with methoxy groups in the natural product.

There can also be acetyl groups present on the free hydroxy

groups. The galacturonic acid main chain also has the
occasional rhamnose group present which disrupts the
chain helix formation. MUCOPOLYSACCHARIDES
Sources of pectin are polysaccharides that contain either an AMINO SUGAR
or a derivative of an AMINO SUGAR
• Citrus fruits contain a large amount of pectin
• Also pears, apples, guavas Hyaluronic Acid is an example of a Mucopolysaccharide
• Typical levels of pectin in plants are (fresh weight):
• It is found in connective tissue and is composed of
 apples, 1–1.5% D-Glucoronic Acid and N-AcetylGlucosamine
 cherries, 0.4% • It helps in blocking the spread of invading
 oranges, 0.5–3.5% Microorganisms
 citrus peels, 30%
 2) BENEDICT’S TEST
Principle:

• Carbohydrates with free aldehyde or ketone groups

have the ability to reduce solutions of various metallic
ions.

• Reducing sugars under alkaline conditions tautomerise

Both conformations (chair and boat) exist for glucose, and form enediols.
though the chair form is thermodynamically more stable • Enediols are powerful reducing agents.

• They reduce cupric ions to cuprous form and are

themselves converted to sugar acids.

• The cuprous ions combine with OH- ions to form yellow

cuprous hydroxide which upon heating is converted to
red cuprous oxide.

Interpretation:
QUALITATIVE ANALYSIS FOR CARBOHYDRATES • Benedict‘s test is a semi quantitative test. The color of the
1. MOLISCH TEST precipitate gives a rough estimate of a reducing sugar
present in the sample.
Principle:
• Green color - Up to 0.5 G% (+)
• Carbohydrates when treated with concentrated • Green precipitate - 0.5-1.0 G% (++)
sulphuric acid undergo dehydration to give furfural • Yellow precipitate -1.0-1.5 G% (+++)
derivatives. • Orange precipitate- 1.5-2.0 G% (++++)
• These compounds condense with Alpha naphthol to • Brick red precipitate- > 2.0 G %(+++++)
form colored products.
• Pentoses yield furfural while Hexoses yield 5-Hydroxy
methyl furfurals.
• An appearance of reddish violet or purple colored ring
at the junction of two liquids is observed in a positive
Molisch test.
3) BARFOED’S TEST • Overheating of the solution should be avoided.
Principle: • Upon continuous boiling, aldoses get converted to
ketoses and give a positive reaction with Seliwanoff
• Aldoses and ketoses can reduce cupric ions even in
reagent.
acidic conditions.
• This test is used to distinguish reducing mono saccharides
from disaccharides by controlling pH and time of 5) HYDROLYSIS TEST FOR SUCROSE
heating.
Principle:
• Mono saccharides react very fast whereas
disaccharides react very slowly. • Sucrose on hydrolysis with HCl is converted to glucose
and fructose.
Interpretation
• The presence of these two monosaccharides can be
• A scanty brick red precipitate is observed in a positive
confirmed by Benedict’s and Seliwanoff test
reaction.
• The positive reaction indicates the presence of a
reducing mono saccharide.
6. OSAZONE TEST
• On prolonged heating disaccharides can also give this
test positive. Principle:
• The solution should be boiled for 3 minutes only
• A solution of reducing sugar when heated with phenyl
hydrazine, characteristic yellow crystalline compounds
4) SELIWANOFF’S TEST called Osazone are formed.
Principle: • These crystals have definite crystalline structure,
precipitation time and melting point for different
• Keto hexoses on treatment with hydrochloric acid form
reducing sugars.
5-hydroxy methyl furfural which on condensation with
resorcinol gives a cherry red colored complex. Interpretation
Interpretation: • Glucose, fructose and mannose produce the same
Osazone because of the similarities in their molecular
• This test is given positive by ketohexoses so it is answered
structure.
by fructose, sucrose and other fructose containing
carbohydrates. • Galactosazone crystals are formed in 7 minutes.
• This test distinguishes between glucose and fructose. • Maltosazone crystals are formed in 10-15 minutes.
7) BIAL’S TEST
Principle:
• The test reagent dehydrates pentoses to form furfural.
• Furfural further reacts with orcinol and the iron ion
present in the test reagent to produce a bluish product.
Interpretation:
• This test is specific for pentoses.
• Hexoses generally react to form green, red, or brown
products
8) IODINE REACTION
• This is a test for polysaccharides
Principle:
• Iodine forms a coordinate complex between the
helically coiled polysaccharide chain and iodine
centrally located within the helix due to adsorption.
• The color obtained depends upon the length of the
unbranched or linear chain available for complex
formation
Interpretation
• Amylose- A linear chain component of starch, gives a
deep blue color
• Amylopectin- A branched chain component of starch,
gives a purple color
• Glycogen- Gives a reddish brown color
• Dextrins- Amylo, Eryhthro and Achrodextrins, formed as
intermediates during hydrolysis of starch give violet, red
and no color with iodine respectively.
• Yellow-orange - negative. Purple-black -positive.
REAGENTS
Iodine solution:
Add a few crystals of iodine to 2% potassium iodide solution till
the colour becomes deep yellow.
Fehling’s reagent A:
Dissolve 34.65 g copper sulphate in distilled water and make up
to 500 mL.
Fehling’s reagent B:
Dissolve 125 g potassium hydroxide and 173 g Rochelle salt
(potassium sodium tartrate) in distilled water and make up to 500
mL.
Benedict’s qualitative reagent:
Dissolve 173 g sodium citrate and 100 g sodium carbonate in
about 500 mL water. Heat to dissolve the salts and filter, if
necessary. Dissolve 17.3 g copper sulphate in about 100 mL water
and add it to the above solution with stirring and make up the
volume to 1 L with water.
Barfoed’s reagent:
Dissolve 24 g copper acetate in 450 mL boiling water.
Immediately add 25 mL of 8.5% lactic acid to the hot solution. Mix
well, Cool and dilute to 500 mL.
Seliwanoff’s reagent:
Dissolve 0.05 g resorcinol in 100 mL dilute (1:2) hydrochloric acid.
Bial’s reagent:
Dissolve 1.5 g orcinol in 500 mL of concentrated HCl and add 20
to 30 drops of 10% ferric chloride.