WELCOME ABOARD FLIGHT BCHEM 153

BCHEM 153 GENERAL BIOCHEMISTRY I

LECTURER (PILOT)
F. O. MENSAH
CO-PILOT (TA): JOHN ACQUAH-MENSAH
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 What is Biochemistry?
• It is the study of the chemistry of life or chemistry of living
things or a laboratory science with a physico-biochemical
focus on all types of cellular activities.

1. As laboratory science – involves inquisitive,

imaginative and patient investigator for successful and
productive research; the availability of proper tools and
the skill to use them depends on the performance of
analytical procedures in the lab and the interpretation of
collected data.

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2. Physico-biochemical – means that complete analysis and
subsequent understanding of living systems depends on
methods employing physical, biological and chemical
principles e.g. production and utilization of energy in a
living cell – physical level – various forms of energy and
interconversions.

• Biological – study of cellular processes to identify those

that require or yield energy and relationship between the
two.

• Chemical – involves flow of energy through the chemical

molecules that participate in the processes.

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3. Living cell

• All living organisms are composed of the same

types of substances that perform the same type
of general functions.

• The major inorganic substance is water which is

the most important substance for life.

• The major classes of organic substances are the

carbohydrates, lipids, proteins and nucleic acids.
4
 Essentially, biochemistry is the study of these
organic biomolecules and the biochemist will
have to address himself to these questions ;

1) How are they synthesized in a living cell?

2) How are they degraded?
3) How is their degradation linked to the
production of useful chemical energy in the
cell?
4) How is the energy produced?
5) How are these substances interconverted?
6) How do they enter and leave a cell?
5
7) How can they be isolated in the lab?
8) How can they be synthesized in the lab?
9) What is their molecular structure?
10)What specific functions do they perform?
11)What is the explanation for how they
function in terms of their molecular
structure?
12)How do genes control the biochemical
individuality of an organism?
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13) How can the genetic information of an
organism be modified by laboratory
manipulation?

14) How do cells (organisms) communicate with

each other on a chemical basis?

15) What are the explanations for the abnormal or

diseased state?

16) How do antibiotics work?

7
17) What biochemical distinctions exist between
normal and cancer cells?

18) How can medical diagnosis be improved by

using precise biochemical criteria?

17) How do hormones regulate the activities of an

organism and in what other ways are organisms
capable of self-regulation?

Ref: Modern Concepts in Biochemistry by Robert C.

Bohinski.
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 The 'TREE OF BIOCHEMISTRY"

Microbiology/ Physiology
Fermentation Biophysics
BRANCHES

Food Science/
Cell Biology
Nutrition

Medicine Genetics/Biotechnology
and Molecular Biology
Organic
Chemistry Pharmacy

BIOCHEMISTRY
STEM
ROOTS

Structural Chemistry of
Chemistry of the Study of Processes and
components of Metabolism substances that
living matter and store and transmit
relationship of bilogical
function to information
structure

14/11/2017 9
 Course Outline
Structure and Function of Molecular components
of the Body

CARBOHYDRATES
i. Definition and classification
ii. Stereochemistry
iii. Polarimetry

Monosaccharides
i. Configuration of monosaccharides
ii. Ring structures of monosaccharides
iii. Reactions of monosaccharides
a. glycoside formation
b. reaction with phosphoric acid
c. oxidation/reduction reactions
d. chain shortening
e. chain lengthening
f. osazone formation
iv. Monosaccharides of Biological Importance
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 Disaccharides
Polysaccharides
i. Storage polysaccharides
ii. Structural polysaccharides
iii. Heteropolysacchrides
LIPIDS
Classification

Fatty acids
Saturated fatty acids
Unsaturated fatty acids
Types of fatty acids
i. Branched fatty acids
ii. Ketohydroxy fatty acids
iii. Cyclic fatty acids
Essential fatty acids
Properties of fatty acids
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 Reactions of fatty acids
i. Ester formation
ii. Salt formation
iii. Hydrogenation
iv. Halogenation
v. Oxidation
Triacylglycerols (TAGS or fats)
Types of TAG (Simple and mixed)
Separation of fatty acids. Fatty acid analysis
Properties of fats
i. Solubility
ii. Melting point
iii. Specific gravity
iv. Hydrolysis
v. Saponification
Chemical constants of fats
i. Saponification number
ii. Iodine number
iii. Acid number
iv. Acetyl number
v. Rm number
Glycerol
Soaps and Detergency 12
Waxes
Phospholipids and Phosphatides
The Eicosanoids
i. Prostaglandins

ii. Leukotrienes

Steroids
Fat soluble vitamins
i. Vitamin A
ii. Vitamins D
iii. Vitamin E
iv. Vitamin K

13
 RECOMMENDED TEXTBOOKS
• Digested notes on Foundation Biological Chemistry
K. NSIAH
• Principles of Biochemistry LEHNINGER
• Biochemistry D. VOET & J. VOET
• Biochemistry C. K. MATTHEWS & K. E.
VANHOLDE
• Biochemistry T. McKEE & J. McKEE
• Biochemistry L. STRYER
• Biochemistry RAUN
• Principles of Biochemistry ZUBAY, PARSON & VANCE
• Lehninger Principles of Biochemistry NELSON & COX
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BIOMOLECULES
All living organisms contain a variety of
macromolecules that perform specific functions
within a cell. They may serve as
• structural components,
• biocatalysts,
• hormones, etc.
The macromolecules or polymers are formed by
the linking together of the smaller molecules
called monomers or building blocks. (These
usually contain C, H, O2 and N2).
Homopolymer - contains same monomer units
Heteropolymer –contains different monomer units
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MACROMOLECULES MONOMER UNITS

Polysaccharides Monosaccharides or
sugar derivatives
Proteins L-amino acids

Nucleic acids Nucleotides

Lipids Fatty acids

16
 Carbohydrates
• Definition - A class of organic compounds
composed of carbon, hydrogen, and
oxygen with the general formula Cn(H2O)n.
They are considered as hydrates of carbon.

• They may also be defined as polyhydroxy

aldehydes or ketones and their derivatives or
as compounds that yield these derivatives on
hydrolysis.

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Types of Carbohydrates
• Cellulose-most abundant carbohydrate, forms the
main supporting structures of plant.
• Starch-reserve food for plants, stored in grains,
tubers and roots which form the staple food and
main energy sources of the developing countries,
e.g. cassava, yam, maize, rice.
• Glycogen-storage form of energy in animals.
• Sugars-found in fruits, flowers and milk;
sweeteners.
• Vitamins - Ascorbic acid, carbohydrate derivative.

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• Glycosides- some are used in medicine
• Anthocyanin and flavonone groups-
contribute to colours in the plant kingdom
• Nucleic acids- make up genetic material
• Mucopolysaccharides - biological
lubricants in the form of saliva, mucus and
synovial fluid.
Industrial processes based on
carbohydrate include textiles, paper,
plastics and alcohol.
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Classification of Carbohydrates
• Monosaccharides- Simplest carbohydrates, contain a
single sugar unit and cannot be hydrolyzed into simpler
forms. They either contain aldehyde (aldoses) or ketone
(ketoses) groups e.g. ribose, glucose, fructose.

• Oligosaccharides- Contain a few monosaccharide units

(2-10) linked together by glycosidic bonds e.g. sucrose,
lactose.

• Polysaccharides- Largest and most complex

carbohydrates. They contain many (more than 10)
monosaccharide units linked together by glycosidic
bonds. Complete hydrolysis yields individual
monosaccharide units. E.g. glycogen, starch and
cellulose
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21
 STRUCTURE AND
STEREOCHEMISTRY
The structure of an organic compound indicates
the following;

1. What atoms are linked to each other i.e. what

is the order of attachment of the atoms?
2. What are the distances between the atoms
that are linked to each other?
3. What are the bond angles?
4. What is the spatial arrangement of atoms?
5. What are the distances between all atoms
including those that are not linked to each
other?
22
• Stereochemistry is the study of the exact
spatial arrangement of atoms within a
molecule i.e. the three dimensional
structure.

• Differences in the structures of the same

molecule in space gives rise to differences
in some physical and chemical properties
of these molecules.

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Isomers
Isomers are chemical compounds with the same
composition, molecular weight, and molecular
formula, but differ in their physical and chemical
properties (Isomerism).

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 OPTICAL ISOMERS
• These are stereoisomers that are mirror
reflections of one another.
• They have the same structure and formula
but different configurations and rotate plane
polarized light in opposite directions but to
the same extent- one to the right and the
other to the left.
• The pair of compounds is called
enantiomers or enantiomeric pair.
• The two members of the pair are designated
D (dextrorotatory) and L (laevorotatory).
25
 Chiral molecules
• A chiral molecule has a central carbon atom with
four different groups or atoms attached to it.

• The spatial orientation of groups around such a

carbon atom is tetrahedral with the carbon atom in
the centre and the four different groups at the
corners of the tetrahedron.

• Such a compound is asymmetric.

• These are molecules that exist as mirror images but

are not super imposable on each other e.g. pair of
gloves, right and left hands.
26
 Chiral molecules
In order to exhibit optical isomerism, a molecule must be chiral.

A A

E C D D C E

B B

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Achiral molecules
• These are molecules that contain a plane
of symmetry and are super-imposable on
their mirror images, e.g. a pair of socks

H H H H H O H

H C C C C H H C C C H

H H H H H H

Butane Acetone

28
Meso compounds
• Molecules that have chiral centres yet are
achiral. They always contain more than one
chiral centre and have a plane of symmetry
and are optically inactive.
• In meso compounds, one half of the molecule
is a mirror image of the other
CH3 COOH

H C Br H C OH

H C Br H C OH

CH3 CO OH
2,3-dibromobutane Tartaric acid 29
Optical activity
• It is the ability of a chiral substance to rotate the
plane of polarized light.
• Polarimeter is an instrument used for measuring
optical activity

Components of a polarimeter
• Light source
• Polarizer
• Sample tube
• Analyzer

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 Polarimeter
angle of rotation

Observer

light mono polarizer sample tube Analyzer, also

source chromatic a polarizing filter
light

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Polarimeter
• Specific Rotation - It is defined as the rotation in degrees
(observed rotation) caused by a solution containing 1g/ml
of optically active substance in a 1 dm tube at a given
temperature (usually 20oC) using monochromatic light of
the D line of sodium at 589 nm.

• It is designated [α].

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[α]DTemp = Observed Rotation (in degrees)
Length of tube (dm) X conc. (g/ml)
= αobs
lxc
Where [α]DT is specific rotation
T is temperature (usually room temperature)
D is spectral line of source of light (sodium D-line)
αobs is observed rotation
l is length of tube in decimetres
c is concentration of substance in g/ml

If concentration of substance is in g/100ml then the formula becomes

[α ]DT = αobs x 100
lxc

If the sample is a pure liquid and there is enough of it, the rotation can
be measured directly. Such a sample is called a neat sample, that is
no solvent. In this case, the concentration is the liquid’s density, d,
so that
[α ]TD = αobs
dxl 33
Racemic mixtures (racemates)
• This is a mixture of equal amounts of an
enantiomeric pair.
• It contains equal numbers of dextrorotatory and
levorotatory molecules so that the net optical
rotation is zero i.e. for every possible orientation
of one enantiomer a molecule of the other
enantiomer would be in a mirror reflection
orientation.
• Cancellation of rotation occurs thus rendering the
mixture optically inactive.
• It is designated by the symbol ± or prefix dl.
34
• The physical properties of racemic mixture may
differ from the pure enantiomers. A racemate may
crystallize in different ways;
• As separate crystals of the (+) and (-) forms, i.e.
the formation of a crystalline substance that is a
simple mixture of the different crystalline forms.

• As one type of crystals containing equal

numbers of (+) and (-) molecules.

• The resulting crystals behave like a separate

compound with higher or lower melting point than
the pure enantiomers.

35
• Racemates therefore often differ from the pure
enantiomers in many physical properties like density
and refractive index.

• The process whereby pure enantiomers are converted

to a racemic mixture is called racemization. This may
occur as a result of chemical reactions.

• Though both meso compounds and racemic mixtures

are optically inactive, a meso compound is a single
molecule whereas a racemic mixture is a 50:50
mixture of an enantiomeric pair. When one enantiomer
is in excess, then a net rotation of plane polarized light
occurs.
36
Monosaccharides
• These are sugar units with the empirical formula (CH2O)n
where n is three or more

• Monosaccharides are classified according to

– The number of carbon units
– Whether they contain aldehyde (aldoses) or ketone
(ketoses) groups

37
The Trioses
These are
Glyceraldehyde – Aldotriose
Dihydroxyacetone – Ketotriose
HCO
H2C OH
H C OH
C O
H2C OH
H2C OH

D-glyceraldehyde Dihydroxyacetone

• While dihydroxyacetone is achiral, glyceraldehyde is chiral

and therefore exists in two enantiomeric forms, D and L.

38
 HCO HCO

H C OH HO C H

H2C OH H2C OH

D (+) glyceraldehyde L-glyceraldehyde

• These two enantiomers serve as configuration

standards for all monosaccharides.

• A monosaccharide whose penultimate carbon or

the highest numbered chiral carbon has the same
configuration as D (+) glyceraldehyde (i.e. OH
group is to the right) is designated as a D sugar.

39
• If the penultimate carbon has the same
configuration as L-glyceraldehyde (i.e. OH group to
the left) it is designated as L-sugar.

• Thus there is no relationship between the D and L

stereochemical families and the direction of
rotation of the compound.

• To illustrate the actual rotation, the prefixes (+) for

dextrorotatory and (-) for laevorotatory are used.

• There can be D(+) and D(-) sugars and L (+) and L

(-) sugars.
40
Characteristics of Monosaccharides
• The carbon skeleton is usually un-
branched
• Each carbon atom except one bears a
hydroxyl (OH) group.
• One carbon bears carbonyl oxygen.
• The carbonyl oxygen may reside on the
terminal carbon atom giving an aldehyde
or at any other position giving a ketone.

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42
Configuration of Monosaccharides
The ball and stick model
A A

C E
D
B D B
E

Line and stick model

a
d e

C or C

b
43
Perspective formula
A A

B C D D C B

E E

Fischer projection formula

a a

b e e b

d d

44
For an Aldopentose

HCO

H OH

H OH

H OH

H2C OH

Disadvantages of the Fischer formula include

• The structure is unstable since the carbon atoms are close and
thus repel each other.
• It is not realistic to obtain such right angled bonds in organic
molecules.
45
Ring structures of Monosaccharides
• Evidence suggests that simple sugars DO NOT
behave as aldehydes and ketones in solution.

• This is because sugars undergo the typical acetal

and hemiacetal reactions of aldehydes and
alcohol.

• Sugars usually exist as cyclic forms in solution or

under physiological conditions.

• In glucose, the carbonyl carbon i.e. C-1 can react

with the OH group at C-5 to give a six sided ring. 46
Hemiacetal / acetal and hemiketal / ketal
formation
A reaction between an aldehyde or a ketone with an alcohol (with
a trace amount of acid) yields a hemiacetal (half acetal) and
hemiketal (half ketal) respectively. Continued reaction of the
hemiacetal or hemiketal with excess alcohol results in the
addition of a second molecule of alcohol, loss of water and the
formation of acetal or ketal.

O OH OR
ROH, H+
+
C R C OR ROH, H R C OR
R H
H H

Aldehyde Hemiacetal Acetal

47
 O
OH OR
ROH, H+
C
R C OR ROH, H+ R C OR
1
R R
1 1
R R

Ketone hemiketal ketal

For sugars,
O
HO
CH
OH
HC OH CH
internal reaction HCOH CH
HC OH
HCOH O
HC OH CH
HC O H
H2C OH
H2C OH
hemiacetal form of
monosaccharide
48
49
• Cyclic acetals and ketals are more stable than the
non-cyclic ones.

• They are usually inert to several reagents, e.g. they

are not attacked by bases, organometallic reagents,
reducing and oxidizing agents.

• The formation of an acetal or ketal can be used to

protect the carbonyl group of an aldehyde or ketone
in order to perform reactions on other functional
groups in the compound.
50
Evidence to suggest that glucose is not a
normal aldehyde;
• Does not give normal reactions of
aldehydes e.g. does not react with Schiff’s
base
• Its optical rotation initially changes with
time and eventually approaches an
equilibrium value when dissolved.
• The carbonyl band is absent in UV or IR
spectra

51
The closing of the hemiacetal ring at C-1 of an
aldose yields a new asymmetric or chiral centre and
will therefore have an influence on the optical
rotation.

In ketoses, the new chiral centre is at C-2.

The new chiral centre is called anomeric centre or

the anomeric carbon and the resulting cyclic
compounds are called anomers.

The two new cyclic sugars are designated α and β

depending on the position of the OH group on
the anomeric carbon.
52
 H

HC OH C O HO CH

HC OH HC OH HC OH

HO CH
O HO CH HO CH O

HC OH HC OH HC OH

HC HC OH HC

H2C OH H2C OH H2C OH

-D-glucose open chain form D-glucose -D-glucose

53
54
Mode of determining α and β-forms

- Draw the ring structure using the Fischer projection

system.

- Compare the configuration of the anomeric carbon (C1

in glucose) with highest numbered asymmetric carbon in
the sugar.

- If the groups on the two carbons are on the same side

(cis) the anomer is α. If the two groups are on the
opposite sides (trans), it is β.

55
 H OH
HO H 2 groups trans
C C

HC OH HC OH

HO CH O HO CH O
2 groups cis

HC OH HC OH

HC HC

H2C OH H2C OH

-D-glucose -D-glucose

56
Mutarotation
The cyclic α and β forms of a sugar in solution are in
equilibrium with each other and can readily be inter-
converted through the free carbonyl specie of the open
chain. The inter conversion of α and β anomers of a sugar in
solution is termed mutarotation.
H

C O

HC OH

D-glucose D-glucose
o HO CH
+19 +112o
o mp=146oC
mp=150 C HC OH

HC OH

H2C OH
57
Haworth Projection Formula
These are ring structures of hemiacetal or hemiketal forms of
sugars based on two cyclic ethers, furan and pyran.

O O

furan pyran

O O

five membered ring six membered ring

58
Formation of the Haworth structure from the
Fischer projection

For any D-sugar, the conversion of the Fischer formula to the

Haworth proceeds as follows;
1. Any group that is directed to the right in the Fischer
projection is given a downward projection in the Haworth.

2. Any group that is directed to the left of the Fischer

projection is given an upward orientation.

3. Terminal CH2OH is given an upward orientation in the

Haworth structure.

For L-sugars, points 1 and 2 are the same but the terminal
CH2OH groups are given a downward projection.
59
Formation of the Haworth structure from the Fischer projection
O
CH CH OH
H2C OH
HC OH HC OH
HO O
HO CH HO CH O
OH
HO CH HO CH OH
HC OH HC OH

H2C OH H2C OH
D-galactose D-galactopyranose

Fischer projections

CH CH OH H2C OH
HO CH HO CH
O
HO CH HO CH O OHHO
HC OH HC OH
OH OH
HC OH HC

H2C OH H2C OH
D-mannose D-mannopyranose
60
Reactions of Monosaccharides
Formation of glycosides
• The hydroxyl group (ROH) or hemiacetal of one sugar
can react with another hydroxyl group (ROH) with the
elimination of water to form an ether linkage or an acetal
(R’-O-R).
CH2OH CH2 OH

O O
CH3OH + OH OH + H2O

OH OH O CH3
HO
OH OH
methyl alcohol D-gluco-pyranose methyl--D-glucopyranoside
(hemiacetal) (full acetal)

61
• Glycosides in general are a large class of
carbohydrate derivatives characterized by the
replacement of the anomeric hydroxyl group with
some other substituents.

They are termed O-glycosides, N-glycosides, S-

glycosides, etc. depending on the atom attached to
the anomeric carbon.

Glycosidic bonds can occur between

monosaccharide units and this forms the basis for
the formation of oligosaccharides and
polysaccharides.
62
In the formation of glycosidic bond, the α or the β
anomer of one sugar can be bonded to any of
the OH groups on the other sugar.

The OH groups are numbered to distinguish

them and the numbering scheme follows that of
the carbon atoms.

63
The notation of the glycosidic bond between the two
sugars specifies;

1. Which anomeric form of the sugar is involved in the

bond formation.
2. Which carbon atoms are linked e.g. 2 α-D glucose
units can be linked in two ways α (1→4) and α (1→6).
H2C OH CH2OH

O O
OH OH

OH
OH O
HO OH

(1-4) glycosidic bond

64
H2C OH

O
OH

OH  1,6 glycosidic bond

O between C1 and C6
OH
CH2

O
OH

OH OH

OH

65
When oligosaccharides and polysaccharides are formed
through glycosidic bonding, their chemical nature
depends on the
• monosaccharide components
• type of glycosidic bond formed i.e. which anomers and
which carbon atoms are linked together.

Example; starch, glycogen and cellulose

Both linear and branch polymers can be formed.

The glycosides are usually resistant to alkali but are

hydrolyzed on boiling with dilute mineral acids.

66
Reaction with Phosphoric acid
The OH groups of sugars behave like other alcohols and
therefore can react with acids and derivatives of acids to
form esters. The sugars form esters with phosphoric acid
to yield phosphorylated derivatives.

H2C OH H2C O P

O O
OH + ATP OH + ADP

OH OH OH OH

OH OH

67
Oxidation – reduction reactions of
Monosaccharides
Action of acids
Monosaccharides are generally stable in dilute acid even on
heating. However in the presence of strong mineral acids
and heat, sugars become dehydrated.
Aldopentoses are dehydrated to form furfurals.

HCO
C
H+ or H2SO4
Pentoses or HC
heat
O CHO O
HC
furfural
HC
68
 Aldohexoses form hydroxymethyl furfural.

HC O
HCOH HCO

HO CH C
+
H or H2SO4
or
HCOH heat HC O
HOCH2 O CHO
HC OH HC
H2C OH hydroxy methyl furfural C
aldohexose H2C OH

69
 The furfurals readily polymerize to give brown tars.
• This dehydration reaction is the basis of certain qualitative
tests for sugars as the furfurals can react with phenolic
compounds such as α-naphthol, resorcinol, and various
activated amines to form characteristic coloured products.
• Example, one of the most important qualitative assays of
carbohydrates is based on the reaction of anthrone in
concentrated H2SO4 with sugars.
• The coloured products can be estimated photometrically.

conc. H2SO4
Sugars heat/anthrone coloured product

anthrone
70
Formation of Sugar acids
Sugar acids are formed by the oxidation of the carbonyl group
or the terminal hydroxyl group and are named accordingly.

Formation of Aldonic acid

Sodium hypoiodite (NaOI) selectively oxidizes aldehyde groups of aldoses
to yield aldonic acid. The reaction is specific for aldoses and is used to
distinguish aldoses from ketoses.
HC O COOH
NaOI
HC OH HC OH
4 4
H2C OH H2C OH

Aldose Aldonic acid

Glucose Gluconic acid

71
 Reaction with bromine water
A mixture of bromine and water is slightly acidic and oxidizes aldoses to
aldonic acid.
Reaction conditions are mild, e.g. room temperature.
HC O
COOH
HC OH Br2/H2O
n HC OH
n
H2C OH
H2C OH

aldose aldonic acid

Examples; for D-allose
HC O C OOH
HCOH HCOH

HC OH Br2/H2O HC OH

HCOH HCOH

HC OH HC OH

H2C OH H2C OH

D-allose D-allonic acid 72

The carbonyl group in the aldonic acid can react with a
hydroxyl group within the molecule to form a cyclic
ester called lactone.
H2C OH C OOH H2C OH
HCOH
O OH  O
Br2/H2O HOCH
OH  OH O
HCOH
 
OH HC OH OH
OH H2C OH OH

-D-glucopyranose D-gluconic acid D-glucono--lactone

The rate of oxidation of the β-D-glucopyranose is about 250

times as fast as the α-anomer.
Aldonic acids exist in equilibrium with their 5 or 6-
membered lactones.
73
• The δ-glucono-lactone may also be formed by aerobic
oxidation of glucose catalyzed by the enzyme glucose
oxidase from Penicillium notatum (enzyme specific for β-
D-glucose.

H
HOC O C

HCOH HCOH
glucose oxidase 
O2 HOCH O
HOCH O + 
HCOH HCOH

HC HC

H2C OH H2C OH

D-glucose D-glucono--lactone

74
Formation of Aldaric acid
Strong oxidizing agents like HNO3 (nitric acid) oxidize both the terminal
alcohol and aldehyde groups of aldoses to dicarboxylic acids called
aldaric acids. Aldaric acids are also called saccharic acids.
HCO COOH
HNO3
HCOH n HCOH n Examples
COOH COOH
HNO3
Aldose Aldaric acid D-mannose D-mannaric acid
HNO3
O O L-ribose L-ribaric acid
C OH C
HNO3
HCOH HCOH D-glucose D-glucaric acid
O
HC OH HC OH

HCOH -H2O HC

HC OH HC OH

C OH C OH

O O
D-aldaric acid lactone of
(from aldohexose) aldaric acid
75
Formation of Uronic acid
Under specific enzyme conditions, the terminal CH2OH groups
alone of aldoses is converted to COOH group to form uronic
acid (also forms lactones).
HC O H CO
enzyme
HC OH HC OH
n n

H2C OH COOH

Aldose Uronic acid

Uronic acids are biosynthetic intermediates in various metabolic
processes. Ascorbic acid (vitamin C), for example, is
biosynthesized by way of glucuronic acid. 76
• Many metabolic waste products are excreted in the urine
as their glucuronate salts.

• In the urine, the glucuronic acid forms glycosidic bond to

various hydroxylated compounds for excretion of these
compounds, e.g. phenols and steroids are excreted as
glucuronides.

• The salts of sugar acids are usually soluble in water.

• The most important ones are glucuronic acid and

galacturonic acid which form part of certain
polysaccharides.

77
 Reducing sugars
• In the presence of oxidizing agents, aldehyde group of
aldoses get oxidized whiles the oxidizing agent is
reduced.

• Aldoses are therefore called reducing sugars.

• Ketoses can also be reducing sugars as they can

isomerize to aldoses.

• Tollen’s and Benedicts or Fehling’s reagents are used in

the lab to detect the presence of reducing sugars. These
oxidize and therefore give positive tests with aldehydes
and with α-hydroxy ketones and hence ketoses.

78
Reaction with Tollen’s reagent
• This is a basic solution prepared by dissolving silver oxide
(Ag2O) in aqueous ammonium hydroxide (NH4OH).
• It therefore uses silver ammonia complex ion Ag(NH3)2+ as the
oxidizing agent.
• In the presence of aldehydes, or reducing sugars, a silver
mirror is deposited on the wall of the test tube.
• Ketones are resistant to oxidation and do not react with
Tollen’s reagent.

RCHO + 2Ag(NH3)2+ + 2OH- RCOO- + 2Ag + 3NH3 + NH4+ + H2O

Silver ammonia
complex
silver mirror

79
 H2C OH H2C OH

O + O
Ag(NH3)2
OH - OH O
OH

OH OH OH
OH OH

Lactone

The commercial manufacture of silver mirrors uses

a similar process.
80
Benedict’s and Fehling’s solutions

These are two reagents that use copper ion as the oxidizing
agent. This is based on the reaction of copper II ions to produce
a red precipitate of cuprous oxide (Cu2O) or copper (I) ions.

RCHO + 2Cu2+ + 5OH- RCOO- + Cu2O + 3H2O

aldehyde from copper carboxylate Brick

(II) sulfate (blue) anion red

• In Fehling’s solution, the copper ion is dissolved in a buffer

containing tartrate ion while in Benedict’s solution the copper
is used with sodium carbonate and citrate buffer. The
reagents are quite basic. Tartrate and citrate ions form soluble
complexes with Cu2+ and thus maintain the copper ions in
solution and prevent it from forming the insoluble form.
81
82
• In the presence of a reducing sugar and heat, the
cupric ions are reduced to less soluble cuprous or
Cu (I) ions and are precipitated as a brick red
precipitate.

• The sugar itself is oxidized to a complex group of

compounds including acids.

• Fehling’s and Benedict’s solutions are bluish due to

the presence of copper II ions.

• Carbohydrates that give positive tests with

Benedict’s or Fehling’s reagents are called
reducing sugars.
83
• Although ketones are not generally easily oxidized, ketoses
are an exception in this case due to the presence of the OH
groups on the carbon next to the carbonyl group.

• They can therefore be converted to aldoses via the enediol

intermediate, and the aldoses are oxidized by the reagent.

H2COH HCOH HC O
Cu2+ Positive test
C O C OH HC OH
(Cu2O formed)
R R R
Ketose Enediol Aldose

84
• All monosaccharides are reducing sugars.
• When two or more monosaccharide units are linked together
through their anomeric centres, the resulting sugar is non-
reducing; e.g. the disaccharide maltose is reducing while
sucrose is not.
H2C OH CH2OH

O O
OH 1 4 OH

OH
OH O
OH OH
non-reducing end
reducing end (ring can open to
(no potential for free C=O
yield free C=O at anomeric carbon)
at anomeric carbon)
85
sucrose
6CH OH 1
2 H2C OH
H
5 O O reducing end of both anomers are
2 HO 5 not free-involved in the glycosidic
4 OH 1 linkage
3
3 4
2
OH O
HO 6CH OH
2
OH
glycosidic bond H O
OH

6 H
6
H2C OH 1
H2C OH hemiacetal or hemiketal
CH2OH positive test with B or F
O OH
O or Tollens
5 HO 2 5 HO 2
4 H O
OH 4 O R
H 3 3
H
HO
HO CH2 OH H
1
acetal or ketal
-D-fructose -D-fructose negative test

86
Chemical application of Benedict’s reagent
• Benedict’s reagent can be used to detect the presence
of excess glucose in the urine (glucosuria).

• People suffering from type I insulin dependent

diabetes mellitus do not produce insulin, a hormone
which controls the uptake of glucose from the blood.

• In such individuals, levels of blood glucose rise too

high such that the kidney is unable to reabsorb the
excess glucose and therefore it appears in the urine.

• Though blood glucose levels can be controlled by

insulin injection, glucose levels in the urine should also
be monitored to ensure that the amount of insulin
being injected is correct. 87
• Benedict’s reagent becomes a useful tool as the
amount of Cu2O formed i.e. the degree of colour
change in the reaction is directly proportional to the
amount of reducing sugar in the urine.

• Thus a brick-red colour indicates a very high

concentration of glucose in the urine.

• Yellow, green, blue-green solutions indicate

decreasing amount of glucose in the urine and a
blue solution indicates an insignificant
concentration of glucose in the urine.
88
Action of Alkali on sugars (keto-enol tautomerism)
• Enols (structure with OH group attached to doubly bonded
carbon) are related to an aldehyde or a ketone by a proton
transfer equilibrium known as keto-enol tautomerism.

• Keto and enol forms of the same compound are called

tautomers and their interconversion, tautomerism.

O OH
tautomerism
R CH2C R' R CH C R'
keto form enol form

89
• Tautomerism is therefore the occurrence of structural isomers
of keto and enol forms of the same compound, which are
interconvertible by a rearrangement involving a shift of a
proton from one hydroxyl group to the adjacent carbon in the
presence of alkali.
• The interconversion occurs via an intermediate called the
enolate ion.

• Under most circumstances, keto-enol tautomers are in a state

of equilibrium.

• The reducing properties of sugars are attributed to the

presence of enediols and the formation of the enolate ion.

• Under higher concentrations of alkali, monosaccharides

undergo oxidation, degradation and polymerization.
90
• When glucose, fructose or mannose solution is
exposed to dilute alkaline for several hours, the
resulting solution on treatment with acid,
contains glucose, fructose and mannose.

• This is because the keto and enol forms of

carbonyl compounds are structural isomers and
the two forms are easily interconverted in the
presence of traces of acids and bases.

91
• Monosaccharides, both aldoses and ketoses and other
carbohydrates containing a free sugar group tautomerize to
form the enolate ion in alkali solution.

The enol forms of these sugars are called enediols as the two
OH groups are attached to the double bonded carbon system.

HC O HO C H HO CH2 HO C H HC O
HCOH COH C O HO C HO CH

HO CH HO CH HO CH HO CH HOCH

HCOH HCOH HCOH HCOH HCOH

HC OH HC OH HC OH HC OH HC OH

H2C OH H2C OH H2C OH H2C OH H2C OH

D-glucose trans-enediol D-fructose cis-enediol D-mannose

92
Reduction of sugars
Sodium borohydride An aqueous solution of sodium borohydride
reduces sugars to corresponding alcohols.
HC O H2C OH
NaBH4,H2O
HC OH HC OH
n n

H2C OH H2C OH

Aldose Alditol

H2C OH H2C OH

C O NaBH4,H2O HC OH

HC OH HC OH
n n
H2C OH H2C OH

Ketose Alditol
93
 reduction
Glucose glucitol or sorbitol

used as a sweetener especially in

special diets required to be low in
sugar

reduction
Fructose mixture of glucitol and mannitol

corresponding to the two possible

configurations at the newly
generated stereogenic center at C2

reduction
Galactose galactitol

reduction
Mannose mannitol
94
Hydrogen and other metal catalysts, e.g. platinum and
sodium amalgam reduce sugars in a similar way to yield
the sugar alcohols or alditols.

HC O H2C OH

HC OH H2, Ni HC OH
n catalytic reduction n
H2C OH H2C OH

Aldose
The products of monosaccharide reduction are called
alditols. Since the alditols lack a carbonyl group, they
do not form hemiacetals and exist exclusively in non-
cyclic forms.
95
Reaction of carbonyl centres

Reaction with hydroxylamine (chain shortening)

• Condensation of aldoses and ketoses with hydroxylamine

yields a group of compounds called oximes.

• Treatment of sugar oximes with a dehydrating agent like acetic

anhydride removes water and converts the compound to a
sugar nitrile with the same number of carbon atoms as the
original sugar.

• Further reaction of the nitrile with ammoniacal silver nitrate

removes one carbon as HCN producing a sugar with one less
carbon atom than the original.

96
 HC O HC N OH
HCOH HCOH

HO CH HO CH

HCOH
+ NH2OH
HCOH

HC OH HC OH

H2C OH H2C OH

D-glucose D-glucose oxime

C N
HC O
HCOH
(CH3CO)2O AgNO3/NH4+ HO CH
D-glucose oxime acetic anhydride HO CH + H2O (basic conditions) + HCN
HCOH
HCOH
HC OH
HC OH
H2C OH
H2C OH
D-gluconitrile D-arabinose
97
Ruff degradation (chain shortening)
This involves
• oxidation of the aldose to an aldonic acid using bromine
water.
• Oxidative decarboxylation of the aldonic acid to the next
lower aldose using hydrogen peroxide and ferric sulphate
(Fe (III) sulphate).
OH
HC O C O

HC OH HC OH HC O
Br2/H2O H2O2
HCOH HCOH
Fe2(SO4) 3
HCOH + CO2
HC OH HC OH HC OH

H2C OH H2C OH H2C OH

D-ribose D-ribonic acid D-erythrose

98
Reaction with hydrogen cyanide (HCN) – (Chain
lengthening) Also called Kiliani-Fischer synthesis
or cyanohydrin synthesis

• Addition of hydrogen cyanide to aldoses yields a mixture of

diastereomeric cyanohydrins with one carbon atom more
than the original sugar.

• On conversion to cyanohydrin, the carbonyl carbon

becomes asymmetric.

• The cyanohydrins may be converted to aldoses through

hydrolysis, acidification, lactonization and reduction.

99
Chain extension of D-glyceraldehyde
HC O

HC OH

H2C OH
D-glyceraldehyde
HCN

C N C N

HCOH HO CH

HC OH epimeric cyanohydrins HC OH

H2C OH H2C OH

Ba(OH)2, H3O+
Ba(OH)2, H3O+

100
 OH OH
C O C O

HCOH epimeric aldonic acid HO CH

HC OH HC OH

H2C OH H2C OH

O O
H2C C O H2C HOC O
epimeric -aldolactone
CH CH CH CH

HO HO HO

Na-Hg, H2O
Na-Hg, H2O
pH-3.5
pH-3.5

H H

C O C O

HCOH HO CH
epimeric aldotetroses
HC OH HC OH

H2C OH H2C OH

D-erythrose D-threose

101
Reaction of monosaccharides with Phenylhydrazine
(osazone formation)
Reaction of an aldehyde or ketone with phenylhydrazine results in
the formation of phenylhydrazone.

H2N NH

C O C N NH
phenylhydrazine

carbonyl compound phenyl hydrazone

(aldehyde or ketone)

• A similar reaction occurs on treatment of aldoses and ketoses

with phenylhydrazine yielding the phenylhydrazone of the
monosaccharide. Further treatment with more phenylhydrazine
yields osazone.
102
 HC O HC N NH
HC N NH
HCOH C N NH
C6H5NHNH2 HCOH
HO CH HO CH H2N
HO CH
+ H2N NH + H2O +
HCOH 2 phenylhydrazine HCOH
HCOH
HC OH HC OH
phenylhydrazine HC OH + NH3
H2C OH H2C OH
H2C OH

D-glucose glucose phenylhydrazone D-glucose osazone

103
• For every mole of sugar converted to osazone, 3
moles of phenylhydrazine are used.

• Osazone formation results in loss of chirality at

carbon 2 but does not affect other chiral centres.

• Due to this, sugars that differ in configuration only at

C1or C2 will give the same osazones.

• Osazones formed from glucose, fructose and

mannose are identical because in the Fischer
projection, these molecules are similar in the
lower 4 carbon atoms.
104
• Hydrazones are soluble and difficult to isolate.

• The osazones are relatively insoluble and form

yellowish crystals at different rates for different sugars
under standard conditions.

• They have high melting points.

• Due to these properties, they have often been used

as derivatives for the characterization of sugars.

105
Monosaccharides of Biological importance
Trioses; Glyceraldehyde and dihydroxyacetone.
• In their phosphate forms, these are readily
interchangeable and occur as key intermediates in
glucose metabolism.
• Their reduction yields glycerol which is used in the
formation of triacylglycerols and other lipids.

106
Monosaccharides of Biological importance

Tetroses; Erythrose in its phosphate form as

erythrose-4-phosphate is an important intermediate
in glucose metabolism.

107
Pentoses; Ribose and deoxyribose are constituents of
nucleic acids.
• The reduced ribose, called ribitol takes part in the formation
of some coenzymes.
• Ribulose is an important intermediate in pentose phosphate
pathway.
• D-arabinose and D-xylose are constituents of glycoproteins.

CH2OH
HC OH

HCOH

HC OH

H2C OH
108
• Hexoses; Glucose, fructose, galactose, mannose

109
 Glucose (Dextrose or grape sugar)

• Widely distributed in nature either in the free or

combined form (eg. lactose, sucrose). Occurs
free in fruits and honey.
• It is the monosaccharide unit of starch, glycogen
and cellulose and can be obtained commercially
by the hydrolysis of starch.
• Principal form of carbohydrate circulating in the
blood and is therefore called blood sugar.
• Blood contains about 60-90mg/100ml (60-
90mg/dL) of blood in the fasting state.
110
Glucose (Dextrose or grape sugar)

111
 Glucose (Dextrose or grape sugar)
• The analytical test for Glucose is most often
performed in a clinical chemistry laboratory is the
determination of glucose in blood, urine, or other
biological fluids.

• Blood glucose levels are measured by an enzyme-

based procedure using the enzyme glucose
oxidase.

• This enzyme catalyzes the oxidation of b-D-

glucose to D-gluconic acid.
112
Glucose (Dextrose or grape sugar)

113
 Functions of Blood Glucose
1. For the production of energy. Excess glucose is
converted to glycogen, mainly in the liver and
later broken down to yield energy to build and
operate tissue machinery.

2. Serves as direct fuel for the brain. (Red blood

cells and the brain utilize glucose exclusively
for their energy needs although the brain in
prolonged starvation also utilizes ketone
bodies).

3. In the mammary gland, some glucose is

converted to galactose and the two link up to
form lactose which is milk sugar.
114
4. The phosphoric acid ester of glucose is a
major source of energy in metabolism.

5. Glucose or some of its derivatives

combine with proteins (glycoproteins) or
lipids (glycolipids) to form important
constituents of cells.

6. Excess glucose can be converted to fat

and stored as fat in fat depots.

115
 Galactose
• It is an epimer of glucose with respect to carbon 4.
• Links up with glucose to form lactose.
• Some yeasts ferment it readily to form galactaric acid or mucic
acid which is optically inactive.
• Inability to metabolize galactose leads to galactosemia and
cataract formation.
• Galactaric acid is a meso compound.
HC O C OOH
HCOH HCOH

HO CH HNO3 HO CH
HO CH HOCH
HC OH HC OH
H2C OH COOH
116
Galactose

117
 Functions of Galactose
1. Can be converted to glucose in the liver
and metabolized, thus serving as a
source of energy.

2. Forms constituents of glycoproteins,

glycolipids and blood group substances.

3. Synthesized in the mammary gland to

make the lactose of milk.
118
 Mannose
• An epimer of glucose with respect to carbon 2.

• Found in nature mainly in the form of polysaccharide

called mannans or mannosans which are complex
polysaccharides that make up gum and rubber
tissues in plants.

• Readily distinguished from glucose by the formation

of sparingly soluble crystallized colourless osazones
when treated at room temperature with phenyl
hydrazine.

• Forms constituents of many glycoproteins.

119
 Fructose ( Levulose or fruit sugar)
• Occurs in fruit juices, honey, and cane sugar,
but not found in large quantities in animals
except under certain conditions.

• Its phosphorylated forms, F-6-P and F-1,6BP

are involved in carbohydrate metabolism of
plants and animals and therefore yield energy.

• On per gram basis, fructose is twice as sweet

as sucrose and can therefore be used in
smaller amounts. 120
Fructose ( Levulose or fruit sugar)

121
Fructose ( Levulose or fruit sugar)

122
• β-form is more abundant and levorotatory.

• Forms the same osazone as glucose with

phenyl hydrazine.

• Undergoes mutarotation (rotation of

freshly prepared fructose is -133.5o but
changes to -92.3o when allowed to stand
i.e. equilibrium specific rotation).

123
 H2C OH H2C OH
CH2OH HO
O O
HO HO

H OH H
HO HO H2COH

-D-fructose -D-fructose

Functions
1. Can be converted to glucose in the body and used as
energy source.
2. Inulin, a polymer of fructose is used in the study of
glomerular function.
3. Often used as sweetening agent in processed food
products because of its sweetness.
124
125
Other derivatives of sugars
Amino sugars
• Sugars in which a hydroxyl group is replaced by an amino
acid or a substituted amino group usually the C2 of aldoses.
Most common are D-glucosamine and D-galactosamine.
• These are prominent components of a large variety of
polysaccharides and other substances found in nature.

CH2OH CH2OH

HO
O O
OH OH

OH OH
OH
NH2 NH2
D-glucosamine D-galactosamine

126
The amino group may bear an acetyl group (CH3CO).

• D-glucosamine, found as component of glycoprotein and

mucopolycsaccharides like heparin and blood group
oligosaccharides (A, B and O), and exoskeleton of insects.
• D-galactosamine is a component of mucopolysaccharides,
chondroitin in cartilage.
• The amino sugars react like hexoses, except that they cannot
form osazones.
127
128
Uronic acid
E.g. D-glucuronic acid and its epimer, L-iduronic acid.
• In liver cells, glucuronic acid complexes with molecules
such as steroids, certain drugs, and bilirubin to improve
water solubility (detoxification).
• Both derived sugars are abundant in connective tissue
carbohydrate components.
Deoxysugars
H replaces an OH group in a sugar, e.g. deoxyribose in DNA

129
Vitamin C (Ascorbic acid)
• It is a derivative of glucuronic acid.
• It is an enediol lactone (ɣ) which bears a double bond
between the second and third carbon atoms.
• It is a white crystalline substance with an acid taste.
Sources; Fresh fruits, green leafy vegetables, citrus fruits

130
Vitamin C (Ascorbic acid)
Properties
• Readily soluble in water but insoluble in organic
solvents.
• It is a strong reducing agent and is easily oxidized in
aqueous solutions in the presence of air to
dehydroascorbic acid.
• (The rate of such oxidation is increased by traces of
copper or an alkaline pH.)
• Relatively stable in acid solution.

131
Oxidation-reduction of ascorbic acid
• Ascorbic acid oxidase (a copper containing enzyme found in
plant tissues) oxidizes ascorbic acid to dehydroascorbic acid
and water. (The D-isomer is biologically inert.) When
dehydroascorbic acid is further oxidized, it is cleaved between
C2 and C3 to give oxalic acid and L-threonic acid.
O C O C
COOH
HO C O C
O 1
/2O2 O COOH HC OH
HO C
ascorbic
O C Further oxidation
and cleavage
+
HOCH
HC acidoxidase HC COOH
HO CH HO CH HOCH2

H2C OH H2C OH
Dehydroascorbic acid oxalic acid L-threonic acid
L-ascorbic acid

The enediol nature of ascorbic acid enables it to take part in

various oxidation-reduction reactions in the body
132
 Biochemical Function of Ascorbic Acid
• It acts as a cofactor for hydroxylation reactions,
e.g. reactions involved in the formation of tyrosine
from phenylalanine, hydroxy proline from proline
and nor-adrenaline from dihydroxyphenyl
ethylamine.

• It is important in the formation of collagen and

therefore functions in growth and repair of
connective tissues, teeth, bones and cartilage.

• It promotes wound healing.

133
• It protects other antioxidants like vitamin E and
A.
• It is a chelating agent and facilitates the
absorption of iron from the intestines.

• It is essential for normal protein metabolism and

deficiency leads to the lowering of the serum
albumin concentration and high rates of nitrogen
loss in the urine.

• It functions in biosynthesis of several hormones.

134
• RDA is 60 mg- Recommended Daily Allowance is the daily
recommended dose of a vitamin or mineral.

• Deficiency leads to a disease or condition called scurvy,

characterized by degeneration of connective tissues.

• Collagen fibers become weak as hydroxy-proline is not

formed resulting in formation of lesions on skin and gum and
weakened blood vessels.

• Symptoms-Bleeding gum, loss of teeth, sore joints and slow

wound healing.

135
 Synthesis of ascorbic acid
• Plants and animals except guinea pigs and
primates can synthesize ascorbic acid from D-
glucose.

• D-glucose is first converted to glucuronic acid and

in a series of reactions converted to vitamin C.

• [L-gulonolactone oxidase is absent from species

that can not synthesize the vitamin.]

136
Synthesis of Ascorbic acid
O
HC O HC O COOH C
HCOH HCOH HO CH HO CH
1
/2O2 +
O
HO CH HO CH NADPH/H HO CH aldono lactonase HO CH
ascorbic glucuronate
HCOH acidoxidase HCOH reductase HCOH H2O HC
HC OH HC OH HO CH HO CH
H2C OH COOH H2C OH H2C OH

D-glucose D-glucuronic acid L-gulonic acid L-gulonolactone

O2
L-gulonolactone
oxidase H2O2

O
O
C
C
HO CH
HO C
O O
O C
HO C
HC
HC
HO CH
HO CH
H2C OH
H2C OH
3-keto-L-gulonolactone
L-ascorbic acid

137
Chemical assay of Ascorbic acid
• Ascorbic acid in acid solutions reduces the blue
solution of 2,6 dichlorophenol indophenol to the
leuco-form.

• The oxidized form of the dye is red or pink in acid

solution and colourless when reduced.

• The method is specific for ascorbic acid alone and

has been used for its estimation.

138
 DISACCHARIDES
• Glycosides containing two monosaccharide units.
• Hydrolysis yields individual monosaccharides.
• Easily hydrolyzed by hot dilute mineral acids but
stable to alkali.
C12H22011 + H2O → 2C6H12O6

• May be a reducing sugar if one of the

monosaccharide units has a free anomeric center.
• Reducing property is lost if anomeric centers of both
sugars are involved in the glycosidic linkage as in
the case of sucrose.
139
 DISACCHARIDES
Formation of Disaccharide

140
In systematic naming of the disaccharide:
• The type of glycosidic bond linking the sugars
should be indicated, i.e.
– If the anomeric hydroxyl group involved in the
linkage is in the α conformation, then the sugar
is an α glycoside (e.g. maltose).

– If the anomeric hydroxyl group involved in the

linkage is in the β position, then the sugar is a β
glycoside (e.g. lactose).

141
• By convention, the non-reducing sugar unit is
displayed on the left, and is named by
converting the ending –ose to –osyl, e.g.
glucopyranose becomes glucopyranosyl.

• The carbon atoms involved in the glycosidic

bond formation should be indicated, i.e. α-1,4 β-
1,4 etc.

• The free anomeric carbon of the second sugar

(the reducing unit) can be in either α or β
positions.

142
 Maltose
• Also called malt sugar and contains two glucose
units and linkage is α 1→4.

• It is obtained from the hydrolysis of starch by

enzyme (salivary and pancreatic amylases,
maltase and glucosidase) and dilute acid.

Properties
• A reducing disaccharide (gives a positive test with
Benedicts, Fehling’s and Tollens solution) as it
has free hemiacetal group.
• Forms osazones with phenyl hydrazine.
• Undergoes mutarotation.
• Sources include germinating cereals and malt.
143
Maltose

144
 Maltose
H2COH CH2OH
O O
HO OH

OH O OH
HO OH
maltose

α -D-glucopyranosyl (1,4)- α-D glucopyranose

α -1,4 glucopyronosyl glucopyranose

145
Isomaltose: CH2OH
Also contains 2 glucose units O
but the linkage is α-1,6. OH
Exists as disaccharide unit in HO
glycogen and amylopectin. HO
O

CH2

O
OH

HO OH
OH

146
Cellobiose
• Contains 2 glucose units and therefore resembles maltose
except that linkage is β 1,4. It is therefore a β-glycoside.

• It is obtained from the partial hydrolysis of cellulose by

enzyme (emulsin, a β-glycosidase) and dilute acids.

Properties
• It is a reducing sugar.

• Undergoes mutarotation.

• Forms osazone with phenyl hydrazine.

147
 CH2OH CH2OH
O OH
O
OH OH
O
HO
HO HO

D-glucopyranosyl-(1,4)--D-glucopyranose
or -(1,4) glucopyranosyl--D-glucopyranose

148
• Though the only chemical difference between
cellobiose and maltose lies in the orientation of the
glycosidic bond, this leads to a great difference in
their nutritional value.

• Maltose can be converted to glucose in the

alimentary canal through the action of the enzyme
maltase which acts specifically on the α-glycosidic
linkages while cellobiose is not acted upon.

• As there is no enzyme in the human body which acts

upon the β-glycosidic linkage of cellobiose and its
polymer, cellulose, they are of no nutritional value to
man.
149
Lactose
• Also called milk sugar. It is formed by mammary glands
and is present in milk. Made up of β–D-galactose and D-
glucose units.

• Linkage is β-1,4 glycosidic linkage.

• It is a β-glycoside.

• Hydrolyzed by dilute acid and enzyme lactase, a β-

glycosidase. (β forms are stable than α forms)

150
 H2COH H2C OH
HO O O
OH OH
O
OH
HO HO
-D-galactopyranosyl-(1,4)--glucopyranose or
 galactopyranosyl, D-glucopyranose
151
Properties
• It is a reducing sugar

• Exhibits mutarotaion and forms osazones with

phenyl hydrazine.

• Not fermentable by yeast but can be converted

to lactic acid by Lactobacillus acidophilus or by
Streptococcus lacticus.

152
Lactose intolerance
• This is a condition in which humans are intolerant to
milk and milk products.

• In some adults, a deficiency of the enzyme lactase

causes lactose levels to rise in the alimentary canal.

• Usually, lactase hydrolyses lactose to its component

monosaccharide units which are absorbed into the
blood stream as the enzyme occurs in the intestinal
villi.

153
• The lactose can be acted on by the lactase of
intestinal bacteria yielding hydrogen gas, CO2
and organic acids.

• These products lead to digestive problems such

as bloating of stomach and diarrhoea.

• Undegraded lactose also gives similar

symptoms.

154
Sucrose
• Also called beet or cane sugar or table sugar.
• It is produced in the leaves and stems of plants.
• Found abundant in sugar cane, sugar beet and
certain palms.
• It is the transportable energy source in the plant
kingdom.
• Component monosaccharide units are α–D-glucose
and β-D-fructose in α1- β2-glycosidic linkage (i.e.
anomeric carbons of both sugars are involved in the
glycosidic bond formation).

155
 • Sucrose is therefore not a reducing sugar and
does not form osazones or undergo mutarotation.

CH2OH
H2C OH
O
O
OH   HO
OH
O CH2OH
HO HO

-D-glucopyranosyl-(1,2)--D-fructofuranoside
D-glucopyranosyl, -D-fructofuranose

156
157
• Sucrose can not be synthesized by animals. Sucrose is
dextrorotatory and can be hydrolyzed by enzymes or
weak acids.
weak acid
sucrose or enzymes glucose + fructose

• After hydrolysis, the previously dextrorotatory sucrose

becomes strongly levorotatory. This is because D-
fructose is more levorotatory than D-glucose

invertase -D-glucose D-fructose

C12H22O11 C6H12O6 + C6H12O6
+66.5o
+52.7o -92.4o
158
Invertase is a β-glucosidase.

• Because of the inversion of the sign of rotation in

the reaction, the process is referred to as inversion
and the 50:50 mixture of fructose and glucose is
called invert sugar.

• Honey is mostly invert sugar as the bees supply

the invertase.

• Sucrose is relatively stable to the action of alkali;

however prolonged boiling with acid phenyl
hydrazine causes hydrolysis and the formation of
osazones.
159
• Sucrose is readily fermented by yeast.

• Sucrose can be used as preservative

because of high sugar concentration
(inhibits the growth of microorganisms).

• It is used as a sweetener.

160
General characteristics of sugars
1. They are usually used for their sweetness.

2. They are soluble in water and readily form

syrups.

3. They form crystals when water is evaporated

from their solution (this is how sucrose is
recovered from sugar cane juice.)

4. They supply energy.

5. They are readily fermented by microorganisms.

161
6. They prevent the growth of
microorganisms at high concentration so
they can be used as preservatives.

7. They darken in colour or caramelize on

heating.

8. Some of them complex with proteins to

give dark colours (known as the
browning reaction).

162
• A very important advance in sugar technology has been
the development of enzymatic processes for the
conversion of glucose to its isomer fructose. Fructose is
sweeter than glucose or sucrose.

• Using starch as starting material, it is possible to

produce sugar syrup with the sweetness and other
properties of sucrose.

• Commonly, corn starch is hydrolyzed to provide the

glucose which is then isomerised.

• The United States produces enormous quantities of corn

and with this technology has become less dependent on
imported sucrose, the availability and price of which can
fluctuate greatly.

163
POLYSACCHARIDES
These contain 10 or more monosaccharide units.
There are two groups; Homopolysaccharides
Heteropolysaccharides
In terms of function, there are two classes;
• Structural polysaccharides like cellulose which
provides the frame work for a rigid structure in plants.
• Storage polysaccharides like starch and glycogen
which serve as the food reserves or energy stores in
plants and animals
• Sugar units present include D-glucose, D-mannose,
D-galactose, D-galactosamine, D-glucosamine, etc.
164
These polysaccharides differ from one
another in the following ways;
• Nature of sugar components.
• Molecular weight.
• Number of the end groups in the
molecule.
• Degree of branching.
• Point of attachment of glycosidic bonds.

165
Starch
• Principal carbohydrate of the diet.

• The storage carbohydrate of higher plants and it

occurs in the form of microscopic granules in roots,
tubers and seeds.

• Heating starch with water causes the granules to

swell and produces a colloidal suspension.
166
• Starch consists of two types of polysaccharides, a straight
chain form, amylose, contains α-1,4 linkages only and a
highly branched form, amylopectin, contains both α-1,4
and α-1,6 linkages.
Amylose

CH2OH CH2OH CH2OH

O O O
OH OH OH

HO O O OH
OH HO n OH
non-reducing end reducing end

167
168
APPREY CHARLES 169
• Both yield maltose on partial hydrolysis and glucose on
complete hydrolysis.

• Most starches contain 10-30% amylose and 70-90%

amylopectin (on the average, a ratio of about 1:3).

• Starches differ in the relative content of amylopectin and

amylose and also vary in the nature of their individual chains
as well as molecular weight.

170
Separation of amylose and amylopectin
• This can be achieved by treating hot starch
dispersion with certain substances such as butanol
(or thymol) which complexes with amylose and
causes it to precipitate in semi-crystalline form.

Amylose
• Linear straight chain polymer of glucose (α-1,4
glycosidic linkages).

• Consists of more than 1000 glucose units.

171
• It has both reducing and non-reducing ends and
exists in a helical coil with 6 residues per turn.

• This results in a compact chain.

• Amylose reacts with iodine to give a deep blue

complex.

• This is because the halide is able to occupy a

position in the interior of the helical coil of glucose
units that is formed when amylose is suspended in
water.
172
Amylopectin

Branched polymer of glucose units.

Contains both α-1,4 (straight chain) and α-1,6

(branched chain) glycosidic linkages.

Branching usually occurs at intervals of 20-25 glucose

units.

Amylopectin forms purple or red colour with iodine.

173
• Amylopectin forms stable starch gel due to
the branches within the molecule.

• This allows it to retain water and resist

water seepage.

• Amylopectin-rich starches are often used by

food manufacturers in sauces and gravies
for frozen foods as they remain stable for a
wide temperature range.

174
• When starch is cooked, it becomes more
soluble in water thus increasing its
digestibility and making it more available for
attack by digestive enzymes.

175
Hydrolysis of starch
1. By acid
H+/H2O
starch glucose
2. By enzyme
enzyme
starch amylase glucose + maltose
Both plants and animals contain enzymes that hydrolyze
starch, e.g. α and β amylases-attack α-1,4 glycosidic
linkages.
α -amylases are found in saliva and pancreatic juice and β-
amylases are found in sprouting grains and malt.
176
• α –amylase
An endoglycosidase that randomly cleaves the
glycosidic linkage well inside the starch molecule
yielding a mixture of glucose, maltose and some
fragments larger than maltose.
• β-amylase
An exoglycosidase. Cleaves successive maltose
units from non-reducing end to yield maltose.

Amylose can be completely degraded to glucose and

maltose units by the two enzymes but amylopectin
is not completely degraded.
177
• This is because the branching units and points near branch
points are not attacked.

• Hydrolysis of starch by these two enzymes alone yields a

residue called limit dextrin.

• Limit dextrin is made up of highly branched glucose units.

• The larger branched limit dextrins give a red colour with iodine
and smaller ones give no colour.

• However, de-branching enzymes or α-1,6 glucosidase

found in both plants and animals cleave the α-1,6 linkages.
This, together with the α and β-amylases can completely
degrade starch.
178
Hydrolysis of starch can be monitored through the
following observation;
– Decrease in viscosity of starch solution.
– Appearance of reducing sugars in hydrolysate.
– loss of starch/iodine colour (achromicity of starch/iodine
reaction)

Some properties of starches

• They are not sweet.
• They form pastes and gels in hot water.
• They provide a reserve energy source in plants
and supply energy in nutrition.
• They occur in seeds and tubers as characteristic
starch granules.
179
Glycogen
• Storage polysaccharide found in animal tissues (contains D-
glucose).

• Has a similar structure to amylopectin except that the

chains are more highly branched (branching occurs for
every 10-12 glucose residues).

• Glycogen is found in animal cells in granules similar to

starch granules in plant cells.

• Absent from brain but present in liver, kidney and muscle.

• It may be isolated from the liver and muscle by treatment

with hot concentrated sodium hydroxide (NaOH) in which it
is soluble and precipitated from solution by treatment with
alcohol.
• It yields a red colour with iodine.
180
In animals the enzyme glycogen phosphorylase cleaves 1
glucose unit at a time from the non-reducing end of a branch
to yield glucose-1-phosphate. This enters metabolic pathways
of carbohydrate breakdown. De-branching enzymes play a
role in complete breakdown of glycogen.

Liver glycogen plays a role in regulation of blood glucose levels.

When blood glucose level is high (after a meal or well fed state),
liver cells take up the excess glucose from blood and convert
it to glycogen for storage, thereby lowering the blood glucose
concentration.

When blood glucose levels fall (starvation/fasting or before

meals), liver cells breakdown glycogen to glucose and release
it into blood stream to raise blood glucose levels.
181
Inulin
• Storage polysaccharide found in the bulbs of many plants e.g.
Jerusalem Artichoke, onions and dahlions.

• Contains about 30 fructose units (33) joined by β 2-1 linkages.

Hydrolyzed into fructose by the enzyme inulinase.

• Clinically, it is used in the inulin clearance test for kidney

function as it is able to pass the renal glomerular membrane
and is not secreted or reabsorbed by the kidney tubules.

• As a result it has been used to measure the rate of glomerular

filtration by intravenous injection (into blood stream) as it is not
metabolized and is eliminated at a constant rate by the kidney.
Used as a means of measuring efficiency.

182
STRUCTURAL POLYSACCHARIDES
Cellulose
It is the most abundant structural polysaccharide made up
of glucose units in β-1,4 linkages. Repeating
disaccharide is cellobiose.
CH2OH CH2OH CH2OH
O O O OH
OH OH OH
O O
HO
OH OH n OH

183
184
• The linear arrangement of β –linked glucose units in
cellulose presents a uniform distribution of OH
groups on the outside of each chain.

• When 2 or more cellulose chains make contact, the

OH groups interact with each other to form hydrogen
bonds.

• If the bonding occurs between several cellulose

chains, it gives a highly insoluble rigid and fibrous
structure leading to the mechanical strength of plant
fibers (fibrils).

• This is ideal as cell wall material for plants.

185
• Cellulose is nutritionally not useful to man as
humans do not have the enzyme cellulase that
can hydrolyze cellulose to glucose units.

• Such enzymes usually attack β-linkages which is

common to structural polymers.

• Ruminants and termites, however can use

cellulose as food source because the symbiotic
bacteria in their digestive systems furnish β-
glucosidases.
186
• On complete hydrolysis with strong acids, it yields
only D-glucose but partial hydrolysis yields
cellobiose.
• It is not attacked by α and β-amylases
• It does not give characteristic reaction with iodine.
• Some properties of cellulose –Acts primarily in
supporting structures in plant tissues.
• Resistant to breakdown.
• Insoluble in cold and hot water.
• Not digested by man so does not yield energy.
• It is an important dietary fibre.

187
Chitin
• Structural material of exoskeleton of insects and the shells of
crustaceans.

• It is similar to cellulose in both structure and function.

• It is a linear homopolysaccharide with β-1,4 glycosidic

linkages.

• Monosaccharides unit is N-acetyl- β-D-glucosamine (this

differs from glucose by the substitution of the N-acetyl amino
group [NH-CO-CH3] for the OH group of C2).

188
Chitin
• Like cellulose, individual strands are held together by hydrogen
bonds. Occurs in cell walls of algae, fungi and yeast.

CH2OH CH2OH CH2OH

O O O
OH OH OH
O O O
HO
HN HN n NH

C O C O C O

CH3 CH3
CH3

189
 HETEROPOLYSACCHARIDES
These are high molecular weight carbohydrate polymers
containing 2-6 different monosaccharides or their derivatives.

They are essential components of tissues where they are

combined with proteins as mucoproteins or proteoglycans.

They are also called mucopolysaccharides.

They are usually gel like or gelatinous, sticky and slippery

substances and are highly viscous.

They form the sticky cementing materials in tissues and are

therefore used for lubrication, e.g. are agar, gum Arabic,
pectin, hyaluronic acid, etc.
190
• There are two types – the acidic
mucopolysaccharides which contain hexosamine
and usually uronic acid with or without sulphate
groups.
• The second type is neutral mucopolysaccharides
which contain hexosamine and a neutral
monosaccharide.
• Examples of acid mucopolysaccharides are;
hyaluronic acid (sulphate free), chondroitin 4 and 6
sulphates, heparin, dermatan sulphate.
• They are all negatively charged.
• In most connective tissues of animals, the acidic
mucopolysaccharides are complexed with protein or
peptide residues as glycoproteins or proteoglycans.
191
• Proteoglycans provide the ground or
packing substance of connective tissues.

• They are capable of holding large

quantities of water and occupying space
thus cushioning or lubricating other
structures.

• This property is due to the large number of

OH groups and negative charges on the
molecules, which, by repulsion, keep the
carbohydrate chains apart.
192
Hyaluronic acid
Contains alternating units of D-glucuronic acid and N-acetyl-D-
glucosamine linked by β-1,3 linkage. The resulting
disaccharide is linked by β-1,4 linkage thus the polymer
consists of alternating β-1,3 and β-1,4 linkages.

- -
COO CH2OH CO O CH2OH

O (1,3) O (1,4) O O
O OH O OH O O
O
HO HO
OH HN OH HN

C O C O

CH3 CH3

repeating diasaccharide in glucuronic acid N-acetyl-D-glucosamine

hyaluronic acid 193
It has a glassy and translucent appearance (Greek
word hyalos for glass).

The repeating disaccharide is called hyalobiuronic

acid.

Hyaluronic acid is found in the synovial fluid of

joints, vitreous humour of the eye and in the
umbilical cord.

It may occur in free form or in salt-like combination

with proteins of tissues and tissue fluids.

In tissues, it acts as cementing substance and

contributes to tissue barriers which permit
metabolites to pass through but resist entry of
bacteria and other infective agents. 194
It forms a highly viscous solution in water and is
hydrolyzed by testicular hyaluronidase or by acids
to yield hyalobiuronic acid.

Complete hydrolysis yields equimolar quantities of D-

glucosamine, glucuronic acid and acetic acid.

Hyaluronic acid differs from other types of

mucopolysaccharides in the following;

• It is unsulphated.
• It is not covalently complexed with proteins.
• It is the only type not limited to animal tissue but
also produced by bacteria.

195
 Functions of Hyaluronic acid
• Forms clear, highly viscous solution that serves
as lubricant in the synovial fluid of joints.

• Gives the vitreous humour of the vertebrate its

jelly like consistency.

• Forms essential components of the extracellular

matrix of cartilage and tendons to which it
contributes tensile strength and elasticity as a
result of its strong interaction with other
components of the matrix.

196
Heparin
It is a heteropolysaccharide found in lungs, liver, thymus, wall
of arteries and blood. It consists of D-glucuronic acid and D-
glucosamine. The amino group and some of the OH groups
are sulphated. It is a blood anticoagulant made in mast
cells and finds wide use in medical practice when it is
necessary to prevent blood clotting. Linkage is α-1,4.
- -
COO CH2OSO - COO CH2OSO3-
3
O O O O
OH OH OH OH

O O O O O
O SO3- HN SO3- OSO3
-
HN SO -
3

2-sulfo-glucuronic acid 2,6-disulfo

glucosamine
repeating disaccharide in heparin 197
It has strong acid properties due to the presence of
the COO- and sulphate groups and readily forms
salts.
Highest negative charge density of any known
biological macromolecule.
The Greek word ‘hepar’ means liver.

Functions;
• It prevents the action of thrombin which catalyses
the conversion of fibrinogen to fibrin.
• It inhibits the conversion of prothrombin to thrombin.
• It stops the clumping together of platelets.

198
Chondroitin sulphates (Greek word
‘chondros’ means ‘cartilage).

• Similar to hyaluronic acid except that the amino

sugar is N-acetyl- β-D-galactosamine with a sulphate
group attached to it.

• This sulphate group forms an ester bond to one of

the OH groups of the N-acetyl-D-galactosamine
usually at C6 but esterification can also occur at C4.

• It contributes to the tensile strength of cartilage,

tendons, ligaments and the walls of the aorta.
199
• The position and number of ester bonds depends on the
source of polysaccharides. There are three types, A, B
and C.

• Type A- chiefly present in cornea, cartilage and adult

bone.

• Type C - found in cartilage and tendons.

• Types A and C are made up of equimolar quantities of

N-acetyl-D-galactosamine, D-glucuronic acid and
sulphuric acid.

• In type A, the sulphate group is esterified to position 4 of

galactosamine and in type C, the sulphate is esterified to
position 6 of the galactosamine group.
200
 - -
COO CH2OSO - COO CH2OSO -
3 3
HO HO
O O O O
O OH O OH O O
O
(1,3) 
OH HN OH HN

C O C O

CH3 CH3

repeating disaccharide in
chondiotin sulfate C

• Type B - Also known as β-heparin or dermatan sulphate (Greek

‘derma’ means skin) is found in the skin, heart valves, aorta and tendons.
It is distinguished from types A and C in not being degraded by testicular
hyaluronidase (found in spermatic fluid).
• Contributes to the pliability of the skin.
• Acid hydrolysis yields D-galactosamine, acetic acid, sulphuric acid and L-
iduronic acid (an epimer of D-glucuronic acid with respect to C5).

• Glucuronic acid may also be present to the extent of 10-20% of the total
uronic acid.
201
 HC O HC O HC O
HC OH HC OH HC OH
HO CH HO CH HO CH
HC OH HC OH HC OH
HC OH HO CH HO CH
COOH COOH H2C OH

D-glucuronic acid L-iduronic acid L-idose

CH2O H
H
O-
SO3O- O
COO
O OH O O

OH HN

C O
and 
CH3
n

repeating disaccharide of
dermatan sulfate

202
 Keratan sulphate (Greek word ‘keras’
meaning horn)
• Repeating unit consists of galactose and
N-acetylglucosamine.
• Depending on tissue or animal source it
may also contain N-acetyl neuraminic
acid, mannose, L-fucose, and
galactosamine.
• It is extensively sulfated on both galactose
and N-acetylglucosamine residues.
• They have no uronic acid and their sulfate
content is variable.
203
• They are present in cornea, cartilage, bone and
a variety of horny structures formed of dead
cells: horn, hair, hoofs, nails and claws.

Functions of other naturally occurring

carbohydrates;
• as determinants of blood groups.
• components of antibiotics and gum secretions
which assist in healing of plant wounds.
• form the protective coating of bacteria
peptidoglycan.

204
205
 LIPIDS
• Lipids are greasy materials found in plants
or animal tissues.

• They are chemically diverse groups of

compounds which are insoluble in water.

• Lipids play diverse roles in an organism

depending on their chemical structure or
nature.
206
• Examples are fatty acids, fats or
triacylglycerols, phospholipids, steroids,
etc.

• The distribution of lipids in plants and animal

tissue vary in quantity and type of lipid.

• Non-depot tissue may contain about 1 – 10%

of total lipids while depot tissue or store-
house may contain large amounts, about
90% of the tissue.

207
Properties;
• They are insoluble or sparingly soluble in water but
soluble in non polar or organic solvents like ether,
hot ethanol, chloroform, acetone, benzene, etc.

• They can be extracted from tissues using the

organic solvents.

• They often occur combined with members of other

classes of molecules, e.g. glycolipids contain
carbohydrate and lipid groups and lipoproteins
contain both proteins and lipid groups.
208
Functions of lipids include the following;
1. Structural components of cell membranes, e.g.
Phospholipids.
2. Major storage form of carbon and energy, e.g.
fats. When completely oxidized, they yield twice
as much energy per gram as carbohydrate and
proteins.
3. They are precursors of other important
substances e.g. prostaglandins.
4. They serve as insulation barriers to avoid
thermal, electrical and physical shock.

209
5.Some fat-soluble vitamins have regulatory or
coenzyme functions and some hormones
are lipids.

6. They serve as protective coatings or

barriers to prevent infection and excessive
loss or gain of water.

7. They contribute to flavour or aroma of most

foods, e.g. short chain fatty acids like C4 -
C10 are responsible for much of the flavour
in butter, milk and cheese.
210
 Classification
• There is no simple internationally accepted
system for the classification of lipids.
• The names of the compounds however fall under
certain categories which consider the component
structure present.
They are mainly classified into;
• Simple lipids,

• Compound lipids and

• Derived lipids.
211
• Simple lipids are esters of fatty acids and
glycerol.

• Compound lipids; These include a lot of

materials that contain other substances in addition
to alcohol and fatty acids.

• Four major types are phopshoacylglycerols,

sphingomyelins, cerebrosides and
gangliosides.

• Derived lipids include any material that can not

be neatly classified as simple or compound lipids,
e.g. steroids, carotenoids, lipid vitamins.
212
 Lipids Constituents/ hydrolytic
products
Simple lipids 1. Acylglycerols H+ Glycerol + fatty acids (fa)
hydrolysis

2. Waxes H+ Alcohol + fa aliphatic or

hydrolysis aromatic (both long chain)

Compound 3. Phosphoacylglycerols “ Glycerol + fa + HPO42- +

lipids choline or ethanolamine
primarily
4. Shingomyelins “ Sphingosine + fa + HPO42- +
choline or ethanolamine

5. Cerebrosides “ Sphingosine + fa + simple

sugar

6. Gangliosides “ Sphingosine + fa + 2-6 simple

sugars (including sialic
acid)
213
• 3 and 4 are also referred to as phospholipids due to the
presence of the phosphate groups.

• 4, 5 and 6 are also referred to as sphingolipids due to

the presence of sphingosine.

• 5 and 6 are also referred to as glycolipids due to the

presence of the carbohydrate or sugar units.

• Sulpholipids- derivatives of sphingosine, having a fatty

acid and a sulphated hexose substituents.

• Proteolipids- complexes of lipids and proteins.

• Steroids- derivatives of
cyclopentanoperhydrophenanthrene.
214
CLASSIFICATION OF LIPIDS

215
CLASSIFICATION OF LIPIDS

216
FATTY ACIDS
• Chemically, these are long chain aliphatic
carboxylic acids.

• Most are monocarboxylic acids containing

linear hydrocarbon chains of variable lengths.

• They usually contain even number of carbon

atoms, C12 to C20.

217
• Almost all naturally occurring lipids contain fatty
acids as principal components and therefore
yield these products on hydrolysis.

• A fatty acid bears a carboxyl group at the polar

end while the non-polar end of the molecule
consists of a hydrocarbon chain.

• Under suitable conditions (pH 7) the carboxyl

group ionizes.

• They are usually unbranched.

218
 -
polar COO
head hydrophilic
n
o
n
p
o
l hydrophobic
a
r
t
a
il

219
Types of fatty acids
There are two types;
1. Saturated (no double bonds) and
2. Unsaturated fatty acids (double bonds
present).
• Fatty acids of animal origin usually have a
straight and saturated hydrocarbon chain
(though some may contain unsaturation).
• Those from plant origin are usually
unsaturated.
220
221
Stearic acid Oleic acid
222
• Fatty acids do not occur in free or
uncombined form in cells but are present in
covalently bound forms in different classes
of lipids.

• They differ from each other primarily by the

chain length and number and position of
double bonds if any.

• These two characteristics specify their

physical properties such as melting point,
solubilities, etc.
223
 Saturated fatty acids
These have the general formula CnH2nO2
where n denotes an even number. E.g.
Acetic acid CH3COOH The essence of vinegar. It is
(C2) produced in wine by
bacteria.
Propionic acid CH3CH2COOH Responsible for the strong
(C3) flavour/aroma of Swiss
cheese.
Butyric acid CH3(CH2)2COOH The cause of odour of rancid
(C4) fat. It is the lowest acid
(lowest C number) isolated
from naturally occurring
fats and oils.
224
• The short chain fatty acids usually have sharp, sour
tastes and unpleasant aroma.

• Acetic, propionic and butyric acids are the end products of

carbohydrate fermentation by rumen micro-organisms of
animals.

• C4-C10 acids are present in butter and some plant fats.

• The bad odour of goats is partially due to the presence of

C6-C10 acids i.e. caproic (C6), caprylic (C8) and capric
(C10).

• Palmitic acid (C16) and stearic acid (C18) are the

predominant fatty acids in plant and animal fat (or in
nature).
225
Unsaturated fatty acids
• These are more abundant than saturated fatty acids
with C16 and C18 being predominant.

• Unsaturated fatty acids have lower melting points

than saturated fatty acids when fatty acids of the
same chain lengths are considered.

• In unsaturated fatty acids with single double bonds

(monoenoic), the double bond always occurs
between C9 and C10.

• In polyunsaturated fatty acids, double bonds occur

between C9 and C10 and any other position beyond
that.
226
• They do not usually have conjugated
double bond system.

• Double bonds are usually separated by

methylene groups.

• The general formula varies depending on

the number of double bonds.

• There are CnH2n-2O2, CnH2n-4O2, CnH2n-6O2,

etc. for fatty acids containing single, double
and triple bonds respectively.
227
• The notation used for fatty acids indicates
the number of carbon atoms and the
number and position of double bonds.
Example;
• 18:0-18 carbon fatty acid with no double
bond.
• 18:1- 18 carbon fatty acid with 1 double
bond. ∆ and superscript are used to
indicate the position of the double bond.
• 18:1 ∆9-denotes C18 monounsaturated
fatty acid with the double at position 9 i.e.
between 9 and 10.
228
A. Saturated fatty acids
Common Systematic Abbreviatio Formula Melting Source
name name n point
(oC)
Capric acid n-decanoic acid 10:0 CH3(CH2)8COOH 31.6 Coconut oil, butter

Lauric acid n-dodecanoic 12:0 CH3(CH2)10COOH 44.2 Coconut oil

acid

Myristic n-tetradecanoic 14:0 CH3(CH2)12COOH 54.0 Nut meg, coconut

acid acid oil
Palmitic n-hexadecanoic 16:0 CH3(CH2)14COOH 63.1 Palm oil
acid acid Common in all
animal and
plant fat
Stearic acid n-octadecanoic 18:0 CH3(CH2)16COOH 70.0 Common in all
acid animal and
plant fat

Arachidic n-eicosanoic 20:0 CH3(CH2)18COOH 75.4 Peanut/groundnut

acid acid oil, coconut
229
B. Some unsaturated fatty acids
Common Systematic name Abbrevia Formula Melting Sources
name tion point
(oC)
Palmitoleic Cis-9- 16:1 ∆9 CH3(CH2)5CH=CH(CH2)7 -0.5 Animal and
acid hexadecenoic COOH vegeta
acid ble fat

Oleic acid Cis-9- 18:1 ∆9 CH3(CH2)7CH=CH(CH2)7 13.4 “

octadecenoic COOH
acid
Linoleic acid Cis,-cis,9,12- 18:2 ∆9,12 CH3(CH2)4CH=CHCH2C -9 Lin seed,
octadecadieno H=CH(CH2)7COOH cotton
ic acid seed
oil
Linolenic All cis 9,12,15 18:3∆9,12, CH3(CH2CH=CH)3(CH2)7 -17 ”
acid octadecatrieno 15 COOH
ic acid

Arachidonic All cis 20:4 CH3(CH2)4(CH=CHCH2)3 -50 Peanut/gro

acid 5,8,11,14eicos ∆5,8,11 CH=CH(CH2)3COOH undnut
atetraenoic ,14

acid
230
Geometric isomerism of unsaturated fatty
acids
• Introduction of double bonds gives rise to cis and
trans forms.
• The two forms differ from each other in physical and
chemical properties.
• The cis form is unstable and more reactive, thus
most fatty acids occur in the cis configuration.
• The cis bond creates a bend in the hydrocarbon
chain.
H3C CH2 CH H3C CH2 CH
7
7
HC CH2 C OOH HOO C CH2 CH
7
7

trans oleic acid mp 45 C o cis oleic acid mp 13.4oC

231
• The cis form occurs naturally but the trans form
can only be synthesized.

• The cis and trans configurations are of biological

importance as organisms can distinguish
between them and use one form in preference to
the other.

• Fatty acids are weak acids with pKa values about

4.5.

pKa=4.5
RCOOH RCOO- + H+

232
Conformation of fatty acids
• Fatty acids occur in both chair and boat
conformations but the zig-zag structure is most
stable. palmitoleic acid 16:1 9

H3C COOH

oleic acid 18:19

COOH

boat chair zig-zag

Linoleic acid 18:29,12

COOH

Arachidonic acid 20:4 5,8,11,14

9,12,15
Linolenic acid 18:3
COOH
COOH

233
Other types of fatty acids are branched, hydroxy and
cyclic fatty acids.
1. Branched fatty acids- there are two types
• Isobranching – branching occurs at the end of the molecule
or at the carbon adjacent to the last or end methyl (CH3)
group. This type of branching is common in waxes than in
fats. H C
H3C CH2 3
or CH2
CH3
H3C
• Ante-branching – branching occurs within the molecule, e.g.
tuberculostearic acid obtained from Tubercule bacilli.
H3C CH2 CH H3C CH2 CH CH2 COOH
7 8
CH3 H3C

C18H37COOH
234
• Hydroxy fatty acids contain a hydroxyl group. E.g.
Cerebronic acid found in cerebrum of the brain. They may also
contain keto groups.

H3C CH2 CH CH2 COOH

n n
HO

• Cyclic fatty acids contain cyclic structures within the

molecule. E.g. Lactobacillic acid in bacteria. Chaulmoogric
acid (from oil of hydrocarpus kuazii) used in the treatment of
leprosy.
O O
HC CH
H3C CH2 HC CH CH2 C OH CH CH2 C OH
5 9 H2C CH2 12
CH2
235
Essential fatty acids
• These are polyunsaturated fatty acids which can not
be synthesized in the body and have to be included
in the diet.

• Mammals can not introduce a double bond beyond

carbon atom 9 of the fatty acid chain.

• As a result linoleate (∆9,12) and linolenic acid

(∆9,12,15) cannot be synthesized by mammals.

• Linoleic acid can be converted to arachidonic acid in

human tissues. 236
• These polyunsaturated fatty acids are found in
high concentration in vegetable oils.

• These essential fatty acids are precursors in the

biosynthesis of physiologically important
substances called prostaglandins.

• These are hormone-like compounds which in

trace quantities have effect on many physiological
functions of the cell, e.g. growth, reproduction,
kidney function, skin integrity, etc.

237
Properties of fatty acids
1. Melting point – This is dictated by the chain length
and the number of double bonds within the
hydrocarbon chain.

Melting point increases with chain length and is

lowered by unsaturation, e.g. stearic acid (18:0) has
a higher melting point than oleic acid (18:1).

This trend is also true for triacylglycerols such that

vegetable oils which generally have a higher
proportion of unsaturated to saturated fatty acids
than animal fats have lower melting points.

238
• The difference is due to the differences in their
structures.
• In the solid phase, saturated fats have a uniform
shape (zig-zag) that allows them to pack closely
together easily in a crystal lattice.
• The closer they fit, the stronger the intermolecular
forces and the higher the melting point.
• The introduction of cis carbon-carbon double bond
in unsaturated vegetable oils creates bends (kinks)
in the hydrocarbon chain, making crystal formation
difficult as the molecules do not fit each other
properly.

239
• The more the double bonds, the more difficult it is
for the molecules to crystallize and the lower the
melting point of the oil (more disorder in the
molecule).
• Trans unsaturation can also fit each other.

2. Boiling point – As melting point, increases as

chain length increases.

Lowered by unsaturation thus unsaturated fatty

acids have lower boiling points than saturated fatty
acids of the same chain length.
240
3. Solubility – Short chain fatty acids between C2
and C4 (inclusive) are very soluble in water but
the solubility decreases with increasing chain
length.

• The long chain fatty acids are almost insoluble in

water because of the non-polar nature of the
hydrocarbon chain.

• They are soluble in non-polar and organic

solvents like ether, benzene, chloroform, etc.

241
Chemical properties/Reactions of fatty acids
• The COOH group, double bonds and other
functional groups in the fatty acids determine
their reactions;
Reactions of the COOH group
1. Ester formation –The carboxyl group of fatty
acids react with alcohols to form esters in the
presence of concentrated H2SO4 or HCl.

O O
H+
R C OH + R'OH R C OR
1
+ H2O

242
2. Salt formation – COOH group of fatty acids ionize and
thus form salts with alkali metals and alkali earth
metals, e.g. NaOH and KOH.

O O
- + H+
R C OH R C O

- NaOH RCOO-Na+ + OH-

RCOO +
RCOOH + NaOH RCOO-Na+ + H2O

243
• The salts of sodium and potassium are
readily soluble in water and the solubility
decreases as the hydrocarbon chain length
increases.

• These salts are called soaps.

• The Na and K salts of the fatty acids are

responsible for the cleansing action and
detergency in commercial soap.

244
• The higher molecular weight salts are less
soluble in water and cause precipitation in
hard water.

• When the salts of the fatty acids are

treated with inorganic acids, the free fatty
acid is released.

RCOONa + HCl → RCOOH + NaCl

245
 Reactions of unsaturated fatty acids
1. Ester formation - as saturated fatty acids but slower
and requires heat, acid or both.

2. Hydrogenation – Hydrogen easily adds on to the C=C

bond of unsaturated fatty acids to form the corresponding
set of saturated fatty acids.
• The hydrogenation usually occurs readily at high
temperature and pressure in the presence of suitable
metal catalyst, e.g. Pt, Ni or Pd. This technique is
employed in the production of margarine.

Pt/H2
Oleic Stearic
Pt/H2
R CH CH CH2 COOH R CH2 CH2 CH2 COOH
246
 Trans fatty acids and margarine
• Plant oils such as corn oil and sunflower oil can be
converted to ‘spreadable’ semisolid substances
known as margarines.

• Margarines can be produced by the partial or

complete hydrogenation of the double bonds in
plant oils.

• In addition to saturating the double bonds of the

fatty acid esters, the hydrogenation process can
also change the configuration of the remaining
double bonds from cis to trans.
247
• The physical properties of these trans fatty acids
are similar to those of saturated fatty acids.

• Dietary trans monounsaturated fatty acids can

increase plasma levels of cholesterol and
triglycerides, and thus their ingestion may
increase the risk of cardiovascular diseases.

• Because of that, many margarines are now

produced from plant oils without hydrogenation
by adding other edible components such as
skimmed milk powder (milk from which the
cream has been removed).

248
3. Halogenation
• Addition of halogen across the double bond
occurs under mild conditions of low
temperature.

• The order of reactivity is F>Cl>Br>I. Usually,

Br2 and I2 are added together as iodine
monobromide (Hans solution).

• Iodine monochloride (Wij’s solution) may also

be used.

249
• Halogenation using iodine alone is also used to
determine the number of unsaturation in a fatty
acid molecule.

It also determines the position of double bonds.

R CH CH CH2 COOH
n
+ I2 R CHHC CH2 COOH
n
I I

250
4. Oxidation – Unsaturated fatty acids may be oxidized
by KMnO4, ozone or atmospheric oxygen.

i. KMnO4

• Oxidation with dilute KMnO4 forms a diol across the

double bond.

• Complete oxidation of the diol under vigorous

conditions yields two compounds of low molecular
weight via the formation aldehyde intermediates.

• In the case of dicarboxylic acid, it is first converted

to semi-aldehyde and then further oxidized to
dicarboxylic acid. 251
 dil KMnO4/OH-
R CH CH CH2 COOH
cold R CHHC CH2 COOH
n n

HO OH diol
complete
oxidation

O O O

R C OH + HO C CH2
n
C OH

dicarboxylic acid

252
ii. Oxidation with ozone

• Addition of ozone to the double bonds forms a

complex ozonide which on hydrolysis with water
yields either an acid or an aldehyde.

• Hydrolysis under reducing conditions (e.g. acid

hydrolysis) yields two aldehydes while hydrolysis
under oxidizing conditions (e.g. alkaline) yields two
acids.

253
 O
H3C CH2 CH CH CH2 C OH
n n

O3
O
H3C CH2 HC O CH CH2 C OH
n n

O O
fatty acid ozonide
OH-/H2O H+/H2O
O O

H3C CH2 C OH H3C CH2 CH

n n
O O O O
+ HO C CH2 C OH
+ HC CH2 C OH
n n

254
iii. Oxidation by atmospheric oxygen
• Atmospheric oxygen oxidizes double bonds in
unsaturated fatty acids to yield several oxygen
complexes, e.g. peroxides, enediols, epoxides,
ketohydroxides, etc.

• These oxygen complexes undergo cleavage at

elevated temperatures to yield acids and
aldehydes of low molecular weight.

• These acids and aldehydes are responsible for

the characteristic odour and flavour of rancid fat.

255
 - -
O CH CH C CH
CH2 CH CH CH2 C OH
n n
O O O OH
peroxide ketohydroxide

C CH -
HC CH HC CH

OH OH O O

epoxide unstable compound

256
Biological oxidation – This occurs in biological
tissues and involves the stepwise removal of two
carbon units in the form of acetyl CoA from the
polar end of the molecule.

• This brings about a gradual degradation of the

long chain fatty acids to shorter ones.

• Energy is released in the process and the

stepwise removal of 2 carbon units is termed β-
oxidation.
257
 Omega (ω) – 3 fatty acids
• Carbon atoms of fatty acids are usually
numbered from the carboxyl carbon (C1).

• The carbon atom adjacent to the carboxyl

carbon (C2) is also known as the α-carbon.

• Carbon atoms 3 and 4 are the β and  carbons

respectively and the terminal methyl carbon
is known as the ω-carbon or n-carbon atom.

258
• n-3 fatty acids are a family of polyunsaturated
fatty acids that have in common a final carbon-
carbon double bond in the n-3 position i.e. the
third carbon from the methyl end of the fatty acid
and are often referred to as ω-3 fatty acids.

Examples are
• Linolenic (18:3 (n−3)all-cis-9,12,15-
octadecatrienoic acid),
• Eicosapentaenoic (20:5 (n−3)all-cis-
5,8,11,14,17-eicosapentaenoic acid)
• Docosahexaenoic (22:6 (n−3)all-cis-
4,7,10,13,16,19-docosahexaenoic acid).

• All the three are essential fatty acids.

259
Alpha-linolenic acid (ALA)

Eicosapentaenoic acid (EPA).

Docosahexaenoic acid (DHA)

260
261
 Dietary sources
1. Fish sources – herrings, salmon,
mackerel, sardines e.t.c.

2. Plant sources – linseed, flaxseed and

their oils.

262
 Biological importance
• They have possible anti-cancer effect
particularly breast, colon and prostate cancers.

• They have been reported to decrease blood

cholesterol thus having protection against
cardiovascular diseases.

• They also help in the improvement of immune

function.

263
 TRIACYLGLYCEROLS (TAG)
• They are the simplest and the most abundant
lipids containing fatty acids as building blocks.

• Also called neutral fats or triglycerides and

are fatty acids esters of glycerol.

• Major components of adipose tissue of

animals and storage depot of plants. Not
normally found in membranes.

264
 O
H2C OH H2C O C R1
O
HC OH HC O C R2
O
H2C OH H2C O C R3

glycerol

Types of triacylglycerols

• They are classified according to the identity and position

of the three fatty acid components esterified to glycerol.

• There are two types; simple and mixed fats.

265
• Simple fats or TAGs; All 3 hydroxyl groups of
glycerol are esterified with the same type of fatty
acid, eg tristearin (tristearoyl glycerol), tripalmitin
and triolein (trioleinoyl glycerol)

O
H2C OH H2C O C CH2 CH3
14
O O
HC OH + 3 H3C CH2 C
14
OH HC O C
O
CH2 CH3
14

H2C OH H2C O C CH2 CH3

14

Tripalmitin

266
• Mixed fats; Have 2 or 3 different fatty acid groups
esterified to glycerol. In naming the mixed fats, the 3
carbon atoms of glycerol are designated 1,2,3 or
α,β,α`.

H2C OH


HC OH + 2 C17H33COOH + C15H31COOH
' oleic acid palmitic acid
H2C OH

O
H2C O C C17H33
O
HC O C
O
C15H31
+ 3H2O

H2C O C C17H33

2-palmito-1,3 diolein or palmito ' diolein

267
 O
H2C OH C17H35COOH H2C O C C17H35
(stearic) O
C17H33COOH HC O C17H33
HC OH
+ (oleic)
C
O
H2C OH C15H31COOH H2C O C C15H31
(palmitic)
-stearo 'palmitolein
-oleo ' stearopalmitin

• Usually the unsaturated fatty acid occupies the

β or C2 position while the saturated ones
occur at C1 and C3 (carboxyl group linked to
glycerol by covalent bonds).

268
• TAGs that are solid at room temperature
are called fats and may contain high
proportion of saturated fatty acids.

• Those that are liquid at room temperature

called oils, contain high proportions of
unsaturated fatty acids.

• TAGs are the major storage and transport

form of fatty acids.

269
• Due to their hydrophobic nature, they form
compact anhydrous droplets within cells and
therefore store more energy than the same
mass of glycogen (glycogen is hydrophilic).

• Fatty acids of TAGs are less oxidized, therefore

release more energy on oxidation (9.3 kCal/gm
of fats compared with 4.1 kCal/gm of
carbohydrates).

• Serve as insulation agent against low

temperatures (TAG stored under skin)-animals
who hibernate like bears.
270
271
 Separation of fatty acids from TAG
1. Treat mixture containing TAG with NaOH or KOH
(saponification). This yields sodium or potassium
salts (soaps) and free glycerol, water and non-
saponifiable lipids (mixture)
O
H2C O C R1 H2C OH R1COO-Na+
O
HC O C R2 + NaOH HC OH + R2COO-Na+
O
H2C O C R3 H2C OH R3COO-Na+
Soaps

R1COO-Na+ + HCl R1COOH + NaCl

272
2. Add ether to the mixture and shake vigorously.
Process partitions the mixture into ether layer
containing the non-saponifiable fraction and
the aqueous layer containing glycerol, soaps,
water and excess NaOH (these are water
soluble).

3. Separate

4. Treat aqueous phase with dilute mineral acid,

e.g. HCl. This releases the free fatty acids.

273
• Since fatty acids are non polar, they may be
extracted with suitable organic solvent like hot
ethanol.

• Various techniques can be used to separate short

chain, saturated and unsaturated fatty acids.

• The best procedure for separation of the different

classes of fatty acids is a chromatographic
technique.
• This separates fatty acids of different chain lengths
and different degrees of unsaturation and
saturation.
274
Separation of fatty acids from a mixture of
lipids Crude lipid mixture including
triglycerides

saponification KOH/NaOH (alcoholic)

glycerol + RCOO-K+ + non-saponifiable fraction (lipids)

Dilute with water and

extract with ether

aqueous phase ether phase

glycerol + RCOO-K+ non-saponifiable fraction -
(sterols, fat soluble vitamins, etc.)
acidify with HCl and
extract with alcohol

aqueous phase organic phase (alcohol)

glycerol + KCl RCOOH (free fatty acid)
275
 Gas liquid chromatography for the detection and
estimation of fatty acid components (fatty acid
analysis)
• It is a very sensitive procedure by which fatty acid
composition of microgram quantities of materials can be
resolved.

• This technique is based on the physical separation of a

moving gas phase by adsorption on a column containing a
non-volatile liquid adsorbent (stationary phase) supported on
an inert solid (e.g. silica gel, ground firebrick coated with
lubricant grease, silicone oil or polyesters of high molecular
weight alcohol with dibasic acid).

• The temperature of the column is maintained at 170-225oC.

276
277
• GLC makes use of volatile materials.

• Therefore the fatty acid must first be converted

to a form that can be vapourized.

• They are first converted to methyl esters.

• A sample of the ester mixture is injected at one

end of the heated column constructed out of a
coiled tube.

• A constantly flowing stream of an inert gas, e.g.

Helium, argon and nitrogen keep the volatilized
esters moving through the column (under
pressure).
278
• The fatty acid component of the gas mixture
is separated or partitioned by virtue of their
different solubilities in the stationary phase.

• The esters emerge out of the column in the

order of falling affinities.

• The ones that are retained least emerge first.

• The emerging esters are detected by the

changes in thermal conductivity or ionization
produced in sensitive detectors.
279
Two types of detectors may be used;
1.The thermal conductivity detectors

• This cell is fitted with a coil of wire with a high

temperature co-efficient and records the heat
conducted away from it when a gas or vapour
passes over the coil.

2. Hydrogen flame ionization detector

• This is more sensitive and amplifies the current

produced when organic materials burn in the
hydrogen flame.

280
• These detectors are connected to
automatic recorders which record peaks
when the different fatty acid fractions pass
over it.

• The area under the peak is proportional to

the concentration of the material.

• Comparison with standard mixtures of

known composition gives a direct
relationship.
281
Properties of fats or TAGs
Solubility
• Generally insoluble in water but those with short
chain fatty acids are sparingly soluble in water, e.g.
butyric acid esterified to OH groups of glycerol.
• Fats are soluble in ether, benzene, chloroform, and
other organic solvents.
• They are slightly soluble in cold ethanol and acetone;
however solubilities in these solvents increase with
increase in temperature.
• Hot ethanol is one of the best solvents for extracting
fats.
282
Melting point
• This depends on the fatty acid composition of the
TAG.

• Saturated and long chain fatty acids have high

melting points and this increases with increasing
chain length.

• Unsaturated and short chain fatty acids have lower

melting points.

• TAGs that are liquid at room temperature have a

high proportion of unsaturation and short chain fatty
acids and those which are solid at room
temperature have a higher composition of longer
chain and saturated fatty acids.
283
• As the proportion of unsaturation or short chain fatty
acid increases, the melting point progressively
decreases.

• Oils (liquids) are converted to fats (solids) by catalytic

hydrogenation which converts the double bonds to
single bonds (margarine preparation).
H2/Ni
Liquid oil solid fat called shortening

• Advantage over butter because it has both poly-

unsaturated and saturated fats.

284
Specific gravity
• Naturally occurring TAG have lower specific
gravity than water and therefore float on
water (fats are less dense than water,
ranges between 0.90 and 0.96g/ml).

285
Physical characteristics
• Pure fats are colourless, odourless and tasteless.

• If these properties appear in the fat, they may be

due to the presence of foreign substances that have
dissolved in the fat, e.g. yellow colour of butter is
due to the presence of the pigments carotene and
xanthophylls.

Optical activity
• Some TAGs exhibit optical activity because some of
them have asymmetric centers, e.g. the mixed
TAGs.
286
Chemical properties
• The chemical properties of fats are dictated by the
type of fatty acids present, e.g. if the fats have
unsaturated fatty acids, they are expected to undergo
addition reactions like halogenation, hydrogenation
and oxidation.

• Since all fats are esters, they are expected to undergo

reactions of esters.

287
• Hydrolysis of fats; Heating fats at high
temperatures and pressure in the presence of a
suitable acid catalyst hydrolyses them to glycerol
and fatty acids.

• Enzymes called lipases in pancreatic juice can

also hydrolyze fats.

O
H2C O C R1 H2C OH R1COOH
O
acid
HC O C R2 + 3H2O
heat/pressure
HC OH + R2COOH
O
H2C O C R3 H2C OH R3COOH

288
Saponification
• Hydrolysis of fats using NaOH or KOH (not water).
Sodium or potassium salts (soaps) of the component
fatty acids are released in the process.

• Fats are insoluble in aqueous medium, therefore the

process is facilitated by the addition of alcohol which
dissolves the fat.
O
H2C O C C17H35 H2C OH
O
HC O C C17H35 + 3NaOH HC OH + 3C17H35COONa
O
sodium stearate
H2C O C C17H35 H2C OH

289
Chemical constants of Fats/Oils
Saponification number:
• Defined as the number of milligrams of KOH or
NaOH required to completely saponify one gram of
fat.
• Since fats are mixtures of glycerides which may
contain different chain lengths of fatty acid, the
saponification number is an index of the average
molecular size or chain length of the fatty acids
present.

• It is inversely proportional to the average chain length

of fatty acid. The shorter the chain length, the higher
the saponification number and vice versa.
290
Iodine number:
• Defined as the number of grams of iodine absorbed by
100grams of fats.
• This measures the degree of unsaturation in a fat i.e.
number of double bonds.

Acid number:
• Defined as the number of milligrams of KOH required for the
neutralization of free fatty acid (ffa) present in 1 g of fat (or
oil).

• It is an index of rancidity and detects the extent of

hydrolysis and liberation of free fatty acid that has
occurred.

• This usually happens when fats are stored for a long time
and exposed to heat and moisture. 291
 Acetyl number:
• Defined as the number of milligrams of KOH
needed to neutralize the acetic acid liberated by
saponifying 1 g of acetylated fat.

H H
O O

R C CH2 C O C3H5
x
+ 3(CH3CO)2O R C CH2 C O C3H5
x
+ 3CH3COOH

OH 3 O C CH3 3
hydroxylated glyceride acetic anhydride O

• It is a measure of the number of OH groups

present in a fat/oil. 292
 Reichert Meisel number (RM
number) or volatile fatty acid
• Defined as the number of milligrams of 0.1 M
KOH required to neutralize the volatile soluble
fatty acids obtained from 5 g of fat which has
been saponified, acidified to liberate the fatty acid
and then steam distilled.

• They are the C4 (butyric) to C12 (lauric) acids.

Butter fat has a high RM number as it contains
more of the volatile or short chain fatty acids or
C4 to C8.
293
• RM for butter fat is 26-33 and that of lard is 0.6.

• RM value is a measure of the volatile water-

soluble acid constituents of the fat.

• The various constants provide information about the

nature of the fatty acid present in the mixture of
glycerides composing a fat.

• Fats may be identified by determining the physical

and chemical properties.
294
Rancidity
• This is a chemical change resulting in unpleasant
odour and taste in fat on storage when exposed
to light, heat, air and moisture.

• The process is known as rancidification and

permits hydrolytic or oxidative changes to occur.

• The reaction involves attack at the double bonds

by peroxide radicals with the formation of unstable
hydroperoxides, which undergo further degradation
to yield keto acids and hydroxy keto acids.

295
• It may also involve hydrolysis of component
glycerides into free fatty acids and glycerol or
mono and di-glycerides.

• This process may be facilitated by lipases,

moisture and heat.

• Besides the unpleasant flavour and taste of

rancid fat, the presence of peroxides
contributes to the destruction of fat soluble
vitamins present in the fat.
296
• Short chain fatty acids released after
hydrolysis are responsible for the rancid
flavour.

• Rancidification may be delayed by use of

antioxidants.

• Examples of naturally occurring ones are

vitamins C and E (prevent oxidation).

297
• Synthetic antioxidants include butylated
hydroxyanisole (BHA); butylated hydroxytoluene
(BHT), propyl galate (PG) and di-t-butyl hydroquinone
(TBHQ).
• Agents like metals which accelerate the
hydroperoxidation of fats are called pro-oxidants,
e.g. are Cu, Fe and Ni.
CH3 HO CH3
H3C
C
CH3
CH3
H3C CH3
C C
CH3
CH3 HO CH3 O CH3

2,6-di-tert-butyl-4-methyl 3-tert-butyl-4-hydroxylanisole
phenol BHT BHA
298
 Glycerol
• It is one of the hydrolytic products of fat
(acid hydrolysis or saponification with
alkali). It is a sweet and heavy liquid with
specific gravity of 1.26.

• It is very soluble in water and ethanol but

insoluble in other organic solvents.

• It can be oxidized with hydrogen peroxide

(H2O2) in alkali medium in the presence of
iron salt to produce a mixture of
glyceraldehyde and dihydroxyacetone.
299
• The product reduces Benedict’s solution;
hence this property is used to test for the
presence of glycerol in a hydrolyzed fat.

H2C OH HC O H2C OH
H2O2/OH-
HC OH Fe salt HC OH + C O

H2C OH H2C OH H2C OH

glycerol glyceraldehyde DHA

300
Dehydration of glycerol
• Heating glycerol in the presence of a
dehydrating agent such as potassium
bisulphate (KHSO4) results in the formation of
an unsaturated aldehyde called acrolein.

• This has an irritating odour.

301
• If fats are heated to sufficiently high temperatures,
they may be broken down with subsequent
formation of acrolein from glycerol which accounts
for the odour of hot fat.

H2C OH HC O

KHSO4
HC OH
Heat
CH + 2H2O

H2C OH CH2
acrolein

302
Uses of glycerol
• It is a powerful solvent for many substances
and therefore used in the manufacture of
cosmetics and pharmaceuticals.

• It has the capability of taking up and holding

water and is therefore used as a moistening
agent, e.g. hair products, toothpaste, etc.

• It is also used as a carrier of antioxidants in

food systems.
303
• In medicine, glycerol is used for lowering
blood pressure.

• It is non-toxic and readily utilized by the

body.

• During metabolism, it is converted to

glyceraldehyde and dihydroxyacetone
which can be phosphorylated for the
release of energy.

304
Soaps and detergents
• Soaps are salts of fatty acids. Those made using
short chain fatty acids (C2-C12) are all water
soluble.

• Those with long fatty acid chains are insoluble in

water because of the size of the hydrocarbon chain.

• Sodium and potassium soaps are soluble in water

while magnesium, lead, and zinc soaps are not.
Soaps are very powerful detergents with unusual
solubility properties.
305
Cleansing action of soap
• The cleansing action of soap is related to the
presence of both water soluble and oil soluble
groups within the same molecule.

• Soap is of little use in either acid or hard water.

• In water, soap ionizes to cation and anion.

• Dirt is considered as grease or an oil droplet.

306
• During cleansing, soap allows the efficient
mixing of oil and water (the two become
miscible).

• This is because soaps form micelles in water.

• These are usually spherical clusters of

carboxylate ions that are dispersed
throughout the aqueous phase.

307
• The carboxylate ions are packed together
with their negatively charged polar head
groups (COO-) at the surface, in contact
with the aqueous environment.

• The hydrocarbon tails are in the interior of

the molecule.

• The sodium ions are scattered through out

the aqueous phase as individual solvated
ions.
308
 H2O
RCOONa RCOO- + Na+
R- non polar tail
COO- - polar head

Structure of a micelle

309
310
• Because the surfaces of the micelles are negatively
charged, individual micelles repel each other and
remain dispersed throughout the aqueous phase.

• Water molecules alone are unable to disperse the

greasy oil droplets of dirt because they are unable
to penetrate the oily layer and separate the
individual particles from each other or from the
surface to which they are stuck.

• Soap solutions are able to separate the individual

particles due to micelle formation.

311
• During the cleansing action of soap, the non-polar
hydrophobic interior of the micelle dissolves the oil
droplet.

• The anions of the micelle confer a negative charge on

the oil droplets causing them to repel each other and
thus enhancing the breakdown of the oil droplet.

• This prevents coagulation and the individual globules

become dispersed throughout the aqueous phase
(emulsion).

• The negatively charged carboxyl groups are balanced

(electrically) by an equal number of positive ions, Na+
or K+.
312
The cleansing action of soap is therefore
due to two properties;
• Micelle formation
• Prevention of coagulation of the micelle by
repulsion.

These two properties are referred to as

detergency.

Soaps stabilize oil emulsions by imparting a

negative charge on the oil droplets and
making them hydrophilic.

313
• Soap solution is basically alkaline and it is this
alkalinity that makes it soapy.

• In the presence of acid (H+), soap loses its

detergency and hence it’s cleansing power
due to the loss of the negatively charged
groups.

• No micelles are formed and therefore

precipitation occurs.

• Soap is therefore not useful in acid solution.

314
• Calcium and magnesium soaps are insoluble in
water and do not emulsify oily substances.

• Sodium soaps are less water soluble than

potassium soaps and are less expensive.

• Ordinary soaps are sodium soaps.

• Potassium soaps are usually found in liquid

soaps and shaving creams.

315
In making soaps;
• dyes are added to make coloured soaps,
• antiseptics (phenols) are added for medicated soaps,
• pumice (an abrasive) is added for scouring soap,
• perfumes can be added for toilet soaps.

Uses of soaps;
• Calcium soaps are used as lubricants.
• Zinc soaps are components of talcum powder.
• Lead, Manganese and cobalt soaps are used in the
paint industry. They hasten the process of drying.

316
Soaps were ineffective in commercial laundering
operations for two reasons;

• The soap scum that formed due to the use of

hard water wasted both time and money in
clean up.

• When dissolved in water, soaps produced an

alkaline solution that destroyed delicate
fabrics.

• Both soaps and detergents are surfactants.

317
 RCOO-Na+ + H2O RCOOH + Na+OH

2RCOO-Na+ + Ca2+ (RCOO-)Ca2+ + 2Na+

Water soluble water insoluble (scum)

+
2CH3(CH2)14COO-Na+ + Mg2+ (CH3(CH2)14COO-)2Mg2+ +2Na

318
Synthetic soaps or detergents

• The cleansing ability of soap has been

improved by modification of the molecule to
form synthetic soaps.

• Such synthetic soaps are not affected by the

hardness or acidity of the water used.

• Besides, their detergent action has a powerful

antiseptic effect.
319
• Synthetic soaps are stable in acid
solutions and do not form insoluble Ca and
Mg salts in hard water.

• Such soaps retain their ionic nature even

at high [H+] ion concentration.

• They have better cleansing properties and

produce a neutral solution when dissolved
in water.

320
Preparation of synthetic soaps
• Catalytic reduction of the carboxyl group of a fatty
acid produces an alcohol which can be sulfated or
sulfonated to form synthetic soaps.
catalytic
RCOOH + 2H2 heat
RCH2OH + H2O

su
n

lf a
ta io

ti o
n

n
lu fo
s

RCH2SO2O-Na+ RCH2OSO2O-Na+
sodium alkyl sulfonate sodium alkyl sulfate

synthetic soaps
321
• The synthetic detergents function in the same way
as soaps.

• They have long non-polar alkane chains with polar

groups at the end. The polar groups of most
detergents are sodium sulfonates and sodium
sulfates.

• The presence of sulfur in detergents stabilizes the

RCOONa group. The non-polar ends are
hydrophobic (lipophilic) and attack the grease or oil
droplet.

• The sulphate or sulphonate end is ionic and is

attracted to the water.
322
• Examples of detergents
H3C CH2 CH2 OSO2O-Na+
n

sodium alkyl sulfate

H3C CH2 CH2 OSO2O-Na+

10

sodium dodecyl sulfate

CH3

H3C CH2 HC
n
SO2O-Na+

sodium alkylbenzene sulfonate

323
Disadvantages of using synthetic soaps
• Some synthetic detergents are not biodegradable
i.e. they are not broken down into harmless
chemicals by microorganisms in the soil, especially
detergents containing branched alkyl chain and
tetrapropylene based sodium alkyl benzene
sulfonate.

H3C CH CH2 CH CH2HC CH2 HC SO2O-Na+

CH3 CH3 CH3 CH3
Tetrapropylene based sodium alkylbenzene sulfonate

324
Types of detergents
There are three types; anionic, cationic and non-ionic.

Anionic detergents
• These include various sulfonates, sulfated lauryl
monoglycerides and the sodium salt of lauryl sulfuric
acid.

• They contain a negative charge on the long organic

chain, e.g. sodium lauryl sulfate and sodium p-
dodecyl benzene sulfonate. (These are
biodegradable).
325
Cationic detergents
• These are quaternary ammonium salts containing
aliphatic chains like cetyltrimethyl ammonium salts.
• They find use as dispersants, emulsifiers, dye-fixing
agents, etc. (used in hair conditioners and
shampoos). They contain a positive charge on long
organic chain.
CH3
CH3
+ +
R N CH3 A - H3C CH2 CH2 N CH3 Cl-
16

H3C
CH3

Alkyl trimethyl ammonium salts R octadecyltrimethyl

may be cetyl or lauryl. A may be ammonium chloride
chloride or CH3SO4
326
Non-ionic detergents
• These do not contain ionic species.

• They are fatty acid esters of sorbitan polyethylene

derivative, e.g. tweens - these are polyethylene
sorbitan fatty acids and SPANS-[these are fatty acid
esters of sorbitol anhydride.]

• Non-ionic detergents find use in hair conditioning and

cosmetics, textile softening, etc.

• Frothing or sudsing is minimal so the non ionic

detergents are well suited for automatic dishwashers.
327
 • In addition, synthetic fibers like polyester
become cleaner when washed with non-ionic
detergents.

H3C CH2 CH2 O CH2 CH2 OH

10 7

a non-ionic detergent

328
• Both soaps and detergents are surfactants
i.e. they lower the surface tension of water.

• This property enables the soap or detergent

solution to soak into textile pores and loosen
and remove grease or dirt more effectively.

• The soaps and detergents clean by a

combination of micelle formation and the
ability to act as surfactants.
329
 WAXES
• These are fatty acid esters of long chain alcohols
other than glycerol.

• The alcohols may be aliphatic or aromatic.

• The long chain alcohols are normally mono-hydroxy

though a few may be di-hydroxy.

• They are soft and pliable when warm but hard when
cold (ie, they are malleable at normal ambient
temperatures).
330
• They are found as protective coatings on
skin, fur and feather and on leaves and
fruits of higher plants.
• They also occur on exoskeleton of many
insects providing a barrier against the
environment.
• They are water repelling solids (insoluble
in water -hydrophobic) and have a melting
point between 60-100oC.
• They have a relatively low viscosity when
melted.
331
 Sources of waxes
Animal source
1. Insects
• Some insects secrete waxes, e.g. beeswax - a
mixture of esters, some free fatty acids, alcohols
and hydrocarbons.
• The esters are palmitic, cerotic (C26) and
mellisic (C30) acids with myricyl alcohol
(C30H61OH).
• The long chain alcohols are usually C26-C34.
• The chemical formula of beeswax is
C15H31COOC30H61.
332
Sources of waxes

333
2. Sperm oil (Spermaceti wax)
• This is wax obtained from the head of
sperm whale and is present with TAGs.
• It contains both saturated (C10-C18) and
unsaturated (C12-C20) fatty acids.
• Alcohols range from C16 to C20.
• May be used as a lubricant.
• It consists principally of cetyl palmitate i.e.
ester of cetyl alcohol and palmitic acid
(C15H31COOC16H33).

334
335
3. Ear wax (known medically as cerumen)
• A yellowish waxy substance secreted in the
ear canal of humans and many other
mammals.
• 60% of ear wax consists of keratin, 12-20%
saturated and unsaturated long-chain fatty
acids, alcohols, squalene and 6-9%
cholesterol.
• Fear, stress and anxiety result in the increased
production of ear wax.
• It protects the skin of the human ear canal,
assists in cleaning and lubrication, and
provides some protection from bacteria, fungi,
insects and water.
336
337
4. Wool fat/wax/grease (lanolin)
• Obtained from sheep.

• It is a mixture of fatty acid esters of the sterol

lanosterol (C30H49OH) and agnosterol (C30H47OH).

• It also contains free and esterified cholesterol.

• It forms protective coating over the wool fibers.

• Lanolin can retain water without much change in

consistency (takes up water without dissolving). It is
therefore used in the preparation of cosmetics and
ointments and moisturizing lotions.
338
339
Plant source
1. Carnauba wax (mp 83-91oC)
• This is a protective coating found on some
leaves e.g., Brazilian palm tree.

• The fatty acids and alcohols of carnauba wax

have even numbered carbons ranging from
C26-C34. Eg. Esters of myricyl alcohol and
cerotic acid.

• It also contains C27 paraffin hydrocarbon.

• It is used in the manufacture of polishes.

340
341
342
 Uses of waxes
• They are used in the production of wax
paper to make it water proof or make it
resistant to staining.

• They are used in shoe polishes, wood

polishes and automobile polishes.

• They are used as coating for many cheeses.

343
• They are also used to make fabrics and leather
waterproof.

• Wax with coloured pigments has been used in

the manufacture of crayons and coloured
pencils.

• They are used in cosmetic industry. E.g. in the

manufacture of lipsticks, mascara, etc.

• Waxes also find use in the pharmaceutical

industries.
344
Structural lipids in membranes
• The major feature of biological membranes is a lipid
bi-layer which acts as a barrier to the passage of
polar molecules and ions. Membrane lipids are
amphipathic, i.e. they contain strongly hydrophilic
and hydrophobic groups.

• There are three general types of membrane lipids.

These are;
– glycerophospholipids - hydrophobic portion composed of
two fatty acids joined to glycerol.
– Sphingolipids - a single fatty acid is joined to a fatty
amine.
– sterols 345
Glycerophospholipids
• These are also called phosphoglycerides.

• Two fatty acids are attached in ester linkage to the

first and second carbons of glycerol and a highly
polar or charged group is attached through a
phosphodiester bond to the third carbon.

• Glycerophospholipids are named as derivatives of

the parent compound phosphatidic acid according
to the polar alcohol in the head group.

346
• The attachment of phosphate at either end
makes glycerol chiral and therefore may
be designated as D or L.

• Not all phosphorous containing lipids are

phosphoglycerides, e.g. sphingomyelin, a
phosphorus containing lipid is not a
phosphoglyceride.

347
348
349
 O

H2C OH O H2C O C R1

HO CH R2 C O CH

H2C O - H2C O -
O O
P P
- -
O O
O O

L-glycerol-3-phosphate L-phosphatidic acid (important

intermediate in the biosynthesis
of phosphoglycerides)
O
g
l fatty acid
O H2C O C R1
y
c or R2 C O CH
e fatty acid
r H2C O
o alcohol
O X
l phosphate ROH P x is contributed by an alcohol
-
O
O
phosphatidyl ester

350
• Fatty acids may differ widely as in TAG
O

H2C O C CH2 CH3 stearyl group

O 16

HC O C CH2 CH CH CH2 CH CH CH2 CH3

7 4

linoleyl group
H2C O
O X
P
-
O
O
phosphatidyl ester

Classes of phosphoglycerides
• There are several classes of the phosphatidyl esters and their
classification depends on the second alcohol esterified to the
phosphoric acid.
351
352
353
• Some of the most important lipids in this
class are
phosphatidyl choline (lecithin),
phosphatidyl ethanolamine (cephalin),
phosphatidyl serine,
phosphatidyl inositol,
phosphatidyl glycerol (cardiolipin).

354
• In all of them, there can be a wide
variation in the nature of the fatty acids
within the molecule.

• The alcohols esterified to the phosphoric

acid is usually an amino alcohol (contains
a positively charged amino group).

• They differ in size, shape, and electric

charge of the polar head groups.

355
Lecithins (Phosphatidyl choline)
• Lecithins are present in membranes of higher
organisms and in egg yolk and soya bean
where they act as emulsifiers.

• They have both metabolic and structural

functions in membranes.

• More than 80% of the phospholipids in the

extracellular liquid layer that lines alveoli of
normal lungs is dipalmitoyl lecithin (has palmitic
acid in both positions 1 and 2).
356
• Dipalmitoyl lecithin is a very effective surface
active agent (surfactant-a substance that lowers
the surface tension of a liquid, normally water,
so that it spreads out over a surface).

• It prevents adherence due to surface tension of

the inner surfaces of the lungs, i.e. dipalmitoyl
lecithin decreases surface tension of the
aqueous surface layer of the lung.

• Its absence from lungs of premature infants

causes Respiratory Distress Syndrome
(RDS).

357
Properties
• They exist as zwitterions with an isoelectric pH of
6.7 or higher.

• They are highly reactive and decompose on

storage.

• They are waxy white substances when pure but on

exposure to air and light, turn brown due to auto-
oxidation of the fatty acids and decomposition.

• They are hygroscopic and mix well with water to

form cloudy, colloidal solution from which they can
be precipitated with acetone.
358
Hydrolysis
• On hydrolysis with acid or alkali, the
nitrogenous base (choline), fatty acids,
glycerol and phosphoric acid are released.

• Certain enzymes called phospholipases

hydrolyze lecithin at various sites of the
molecule.

359
• Hydrolysis by phospholipase A obtained from
snake venom, bee stings and pancreatic
juice, yields lysolecithin which has a
powerful hemolytic action on red blood cells.

• At high concentrations, lysolecithin and other

lysophospholipids act as detergents,
dissolving the membranes of erythrocytes and
lysing the cells.

• Extensive haemolysis may be fatal.

360
• Phospholipase B (A1 and A2)-a mixture of
two enzymes) split off both fatty acids from
the molecule.

• Phospholipase C from plant kingdom,

certain venoms and animal tissues splits
lecithin into a 1,2 diacylglycerol and
phosphoryl choline.

• Phospholipase D, from cabbage liberates

choline, leaving a phosphatidic acid.
361
 phospholipase A1

phospholipase A2 O

O H2C O C R1

R2 C O CH
CH3
H2C O +
O CH2 CH2 N CH3
P
phospholipase A - CH3
O
O
phospholipase C
phospholipase D

362
Cephalins (Phosphatidyl ethanolamine)
• The primary amino group in cephalins is less basic
and hence more acidic compared to lecithin.

• They also have a lower solubility in alcohol than

lecithin.

• Phosphatidyl serine is the most strongly acidic

member due to the presence of COO- group.

363
 O
+
HO CH2 CH2 NH3 H2C O C R1
O

R2 C O CH
ethanolamine
H2C O +
O CH2 CH2 NH3
P
-
O
O
phosphatidyl ethanolamine (cephalin)

364
Sphingolipids
• Sphingolipids are another class of lipids
which do not contain glycerol but contain a
long chain amino alcohol with a long
unsaturated hydrocarbon chain (aliphatic
base) called sphingosine.

• They are important membrane components in

both plant and animal cells, especially in large
amounts in the brain and nerve tissues.

365
• They contain three characteristic building
block components; one molecule of long
chain fatty acid, one molecule of
sphingosine or its derivative
dihydrosphingosine, and a polar head
group or amino group.

• Carbohydrate residues in sphingolipids

include glucose, galactose, N-acetyl
hexosamine and sialic acid-
sphingoglycolipids.
366
• The simplest classes of sphingolipids are the
ceramides which consist of one fatty acid
linked to the amino group of sphingosine by an
amide bond.

• Ceramide is a precursor to both sphingomyelin

and cerebrosides.

367
 +
+ OH NH3
OH NH3
H3C CH2 CH2 CH2 CH CH CH2OH
H3C CH2 CH CH CH CH CH2OH 12
12

sphingosine dihydrosphingosine

O
OH HN C R
H3C CH2 CH CH CH CH CH2OH
12

ceramide

368
Sphingomyelins

• In sphingomyelins, the primary alcohol group of

sphingosine is esterified to phosphoric acid, which in
turn is esterified to amino alcohol, choline.

• It is the only phosphorous containing sphingolipid.

• It is found abundant in the brain and nerve tissues.

369
 O
OH HN C R CH3
H3C CH2 CH CH CH CH CH2O +
12 O CH2 CH2 N CH3
-
P
O CH3
O
a sphingomyelin

370
Cerebrosides
• These contain one or more neutral sugar residues as
their polar head groups, e.g. galactose and glucose.

• They do not possess an electric charge and form most

part of the brain’s white matter and myelin sheath.

• Cerebrosides of brain and nervous tissues contain D-

galactose (galactocerebrosides).

• Those in non-neural tissue usually contain glucose

(glucocerebrosides).
371
 O
sphingosine OH HN C R long chain fa
H3C CH2 CH CH CH CH CH2
12

HO CH2 O
HO
O
OH
galactose

H
OH

galactocerebroside
372
Gangliosides

• Complex sphingolipids that contain sialic

acids (neuraminic acid) in addition to glucose
and galactose.

• Found in high concentration in nerve tissues,

spleen and gray matter of the brain.

373
• The sugars and sugar acids may be joined
in branches of about seven (residues) to
form an oligosaccharide which is esterified
to the terminal alcoholic group of
sphingosine.

• Found abundant in nerve endings and have

been suggested to function in the
transmission of nerve impulses across
synapses.

374
 THE EICOSANOIDS
• These are a group of hormone-like molecules
that are produced in most mammalian
tissues.

• They include the prostaglandins,

thromboxanes and leukotrienes.

Prostaglandins
• These are lipid soluble acidic materials which
were first isolated from seminal fluid and the
prostate gland.
375
• They have since been found to occur in many
tissues in the body.

• They are derived from arachidonic acid.

• They are C20 fatty acids containing one or

more double bonds, hydroxyl and keto
groups and a cyclopentane ring involving
carbon atoms 8 to 12 in the middle of the
chain.

376
Types of prostaglandins
• There are different classes of prostaglandins and
these differ from one another in the number and
position of double bonds and oxygen containing
functional groups.

• They are classified as belonging to the series A, B, E

and F and further oxidation products termed G, H
and I series.

• Thus there is PGA, PGB, PGE and PGF.

• Some of the E and F series are widely distributed.

377
• A subscript is attached to the letters to indicate the
number of C=C bond outside the ring.

• The E type has a keto group in position 9 whereas

the F has an OH group in this position.

• The E and F were the first two to be isolated and

were designated so because of their preferential
solubility in ether (E) and phosphate buffer (F) from
fosfat, a Sweden word.

378
Characteristics of prostaglandins;
They possess

• 20 carbon atoms.
• a cyclopentane ring.
• two aliphatic side chains.
• a double bond between carbon 13 and 14.
• a terminal carboxyl group.
• an OH group at position C15.

379
 O O

C OH 7 5 3 C OH
9 8 1
6 4 2
CH3 10
16 18 CH3
1415 20
11 12 13 17 19

Arachidonic acid Prostanoic acid

O O
O OH
C OH C OH

CH3 CH3

OH OH OH OH

PGE1 PGF1

380
O O OH O
C OH C OH
CH3 CH3

OH OH OH OH

PGE2 PGF2

O O
O O
C OH C OH

CH3 CH3

OH OH

PGA1 PGB1

381
Functions
• They modulate the action of hormones rather than
act as hormones themselves.

• They regulate blood pressure (lower blood

pressure).

• They stimulate smooth muscles to contract

(especially uterus, inducing delivery of the foetus).

• They have effect on renal blood flow, gastric

secretions, Na+ uptake, inflammatory reactions, etc.
382
• They have sedative effects on the central
nervous system (CNS).

• They are known to inhibit the aggregation of

blood platelets.

• They may therefore be of therapeutic value in

preventing the types of stroke and heart attacks
that arise from the blood clots which block blood
supply to the brain or to the heart.

• They induce inflammation.

383
• Aspirin inhibits prostaglandin biosynthesis
(by interfering with the oxygen adding
dioxygenase).

• This property of aspirin accounts for its anti-

inflammatory and fever reducing properties.

384
Leukotrienes
• These are also derivatives of arachidonic acid. They
are found in leukocytes or WBCs and have three
conjugated double bonds, hence their name.
O
-
H2C CH C O
cysteine
+
S NH3

COOH
OH
triene
CH3

Leucotriene C
385
Functions
• They induce constriction of smooth muscles
(especially muscles lining the airways to the lungs).

• Asthma attacks may result from this constricting

action, since the synthesis of Leukotriene C appears
to be facilitated by allergic reactions such as
reaction to pollen.

• Drugs that inhibit the synthesis of leukotriene C,

e.g. prednisone (glucocorticoid) are used for the
treatment of asthma.

386
• The strong contraction of the smooth
muscles of the lung that occurs during
anaphylactic shock is part of the potentially
fatal allergic reaction in individuals
hypersensitive to bee stings, penicillin or
various agents.

• They have inflammatory properties and

may be involved in rheumatoid arthritis.

387
 STEROIDS
• These are derivatives of a fused reduced ring system
called the cyclopentanoperhydrophenanthrene nucleus.

• It is made up of three fused cyclohexane rings designated

A, B and C as in phenanthrene and a terminal
cyclopentane ring designated D.

12
13 17
11
C D 16
1
10 14
2 9 15
8
A B
3 5 7
4 6
388
CHOLESTEROL
389
• Molecules with this basic structure are
called steroids.

• Many different steroids with distinctive

functions and activity occur in nature.

• They differ in number and position of

double bonds and in the location,
number, and types of substituent
functional groups.
390
• The main points of substitution are; C3 in ring A, C11
in ring C, and C17 in ring D. The presence of a
hydrocarbon side chain with 8-10 carbon atoms at
C17 and an OH group at C3 characterizes a large
group of steroids called sterols (steroidal alcohols).

• Steroids are derived from the linear triterpene called

squalene which is a C30 hydrocarbon made up of 6
isoprene units.
CH3
or
H2C C CH CH2

isoprene unit 391

• Cyclization of squalene produces lanosterol
which is converted to cholesterol, the most
abundant sterol in animal tissues.

Isoprene → Squalene→Lanosterol →Cholesterol

• Cholesterol and lanosterol are sterols. They

either occur free in nature or esterified with fatty
acids at OH group of C3 (cholesteryl esters).
They are solids at room temperature.
392
Cholesterol
• It is found abundant in animal cells, especially
nervous tissues. It is insoluble in water but
soluble in ether, benzene, chloroform and hot
alcohol.

• It is a major structural component of plasma

membrane and plasma lipoproteins (LDL,
VLDL, HDL).

• It varies in concentration from 0-40% of total

membrane lipids.
393
• Often esterified to fatty acids. It is the
parent compound of all steroids
synthesized in the body, e.g. bile acids,
glucocorticoids, sex hormones and vitamin
D.

• It is present in human serum at

concentration of 160-260 mg/100 ml. Its
deposition in blood vessels results in
artherosclerosis, a condition that
eventually leads to heart diseases.
394
Squalene to cholesterol

squalene

squalene

CH3

CH3

HO
H3C CH3
lanosterol

cholesterol
HO

395
396
Bile acids
• These are C24 sterols synthesized in the
liver from cholesterol. They are the main end
products of cholesterol metabolism.

• Examples are cholic acid and deoxycholic

acid.

• They aid in the digestion and absorption of

fats and are components of bile.

397
• The bile acids are usually conjugated with
glycine and taurine.

• The salts of these conjugated acids are

water soluble and powerful detergents for
fats, facilitating their hydrolysis by lipases
and absorption in the intestines.

398
 OH
OH -
COO C NHCH2CH2SO3H
O

taurine

H2NCH2CH2SO3H HO OH
HO OH
H
H
cholic acid Taurocholic acid
(cholate) (taurocholate)
H3N+CH2COO-
glycine

OH OH
-
C NHCH2COO- C O
O O

HO OH HO
H H

glycocholic acid
Deoxycholic acid
(glycocholate)
399
Unsaponifiable lipids
• These do not contain fatty acids and are found in
smaller amounts in cells and tissues.

• They are broadly classified as isoprene units.

• Examples are the carotenoids e.g. β–carotene (a

precursor of vitamin A or retinol), squalene
(precursor of cholesterol) and the fat soluble
vitamins A, D, E and K.

400
 FAT SOLUBLE VITAMINS
Vitamin A (all-trans-retinol)
• It is an isoprenoid alcohol important for vision, growth
and reproduction in rats.

• Carotenoid pigments in plants act as precursors to

vitamin A, e.g. α, β and γ–carotene with β–carotene
being the major precursor.

• The α– and γ–forms have open rings. Animals

consume these pigments and bring about chemical
alterations to yield vitamin A.
401
• The conversion is enzymatic and occurs in
the liver and intestines.

• β-carotene is a symmetrical molecule and

yields two molecules of retinol/mole of β –
carotene administered. α and γ carotenes
yield one mole of retinol while lycopene
and xanthophylls have no vitamin A
activity. β–carotene is a major carotenoid
in green plants.

402
Sources (carotenoids and vitamin A)- green leafy
vegetables, carrots, brightly coloured fruits and
vegetables, cod liver oil, egg yolk, milk, animal fat,
liver (the richest source, about 95%). Colostrum
also contains a high concentration.

Structure and properties

• There are two types of vitamin A designated A1 and
A2 with A2 being more active.
• A1 or retinol 1 is common in mammalian tissues
and marine species and A2 or retinol 2 is more
common in fresh water species.
• Retinol is a highly unsaturated alcohol with a β-
ionone ring in the structure.
403
• It is insoluble in water though it is possible to
synthesize water soluble derivatives.

• It is stable to high temperatures but is readily oxidized

in the presence of oxygen and trace elements like
copper and cobalt.

• Oxidation can be inhibited by antioxidants like vitamin

E and hydroquinone.

• It is not affected by ordinary cooking and canning

processes.

• Retinyl esters are more stable than the free vitamins.

404
• The aldehyde form, retinal is more reactive
than vitamin A because of the carbonyl group
attached to conjugated double bonds.

• It reacts with amino and sulfhydryl groups of

proteins.

• Absorption of retinol and β –carotene occurs

in the small intestines. Bile salts are required
for their absorption while mineral oils interfere
with their absorption.
405
H3C CH3 CH3
CH3
15
1 10 14 CH2OH
2 6 7 8 9 11 12 13

3
5 CH3
4 C20H29OH

all-trans retinol - vitamin A1

H3C CH3 CH3

CH3
CH2OH

CH3
C20H27OH

retinol-2 - vitamin A2
(3-dehydroretinol)

406
Cleavage of provitamin A

cleavage
H3C
H3C CH3 CH3 CH3

CH3 CH3 H3C CH3

CH3
enzyme
action in [O] carotene (provitamin A)
liver

H3C CH3 CH3

CH3
CH2OH

CH3 H3C CH3 CH3

CH3
CH O
retinol (vitamin A)

CH3
all-trans retinal
407
Biochemical Function-The Photochemistry of
Vision
• Vitamin A plays an important role in vision. There are
two types of light receptors in the retina of the eye,
the cone cells and the rod cells.

• The rod cells cover a fairly large area of the retina

and are mainly found at the periphery. They are
responsible for vision in dim light.

• The cones are mainly found in the central area and

are responsible for colour vision and vision in
bright light (and contribute to acuity of vision).
408
• Hens and pigeons have vision only in
bright light and mainly have cones in their
retina. The owl which hunts at night has
mainly rods in its retina.

• The chemistry of vision has been more

extensively studied in the rod cells than in
cone cells.

• A protein called opsin is essential for the

biochemical function of vitamin A.
409
• The perception of light by rods and cones is
mediated by photosensitive pigments called
rhodopsin (for rods) and iodopsin (for
cones).

• These are protein conjugates.

• Rhodopsin is a brilliant-red heat-labile

protein which is insoluble in water and
bleaches on exposure to light.

410
• Retinol from the liver is transported in the plasma to
the retina bound to retinol binding protein (RBP-mw
21,000-22,000). In the retina, under the action of the
enzyme retinol dehydrogenase, it is oxidized to all-
trans retinal.

• The all-trans retinal is immediately isomerised to 11-

cis-retinal which spontaneously complexes with opsin
to form rhodopsin which is stable but photosensitive.

• All-trans-retinal can not bind to opsin (Rhodopsin

forms about 50% of the protein of the rod cells).

411
The biochemical conversion of retinol to 11-cis retinal

H3C CH3 CH3 H3C CH3 CH3

CH3 CH3
CH2OH CH O

retinol all-trans retinal

retinol dehydrogenase
CH3 CH3

retinal isomerase

H3C CH3
CH3
11

Rhodopsin
opsin
CH3 H3C

CH
11-cis retinal (visual pigment) O

412
• On the absorption of light, the 11-cis retinal of
rhodopsin is isomerised to all-trans retinal
which is readily hydrolyzed to all-trans retinal
and free opsin.

• As a result of this reaction, an electrical impulse

is generated in the optic nerve and transmitted
to the brain to be processed as a visual event. It
is the initial reaction that requires light energy
and all subsequent changes can occur in
darkness or in light.
413
• To regenerate rhodopsin, the all-trans-
retinal is reduced to all-trans-retinol by
alcohol dehydrogenase and NADH.

• This is then isomerised by an isomerase to

11-cis-retinol and followed by oxidation in
the presence of alcohol dehydrogenase
and NAD+ to 11-cis-retinal which
complexes with opsin to form rhodopsin.

414
all-trans retinal all-trans retinal
NADH retinal
alcohol dehydrogenase OR isomerase
NAD+
all-trans retinol 11-cis retinal

retinol
isomerase

11-cis retinol
NAD+
alcohol dehydrogenase
NADH
11-cis retinal

U.V.

Rhodopsin (11-cis retinal) Opsin + all-trans retinal

415
Visual Cycle
Light

Rhodopsin

Opsin
11-cis-retinal all-trans-retinal

enzymatic

416
• When rhodopsin absorbs light, the cis
retinal isomerizes to a trans form which
dissociates from the opsin.

• The dissociation increases the

permeability of rod cell membrane to ions.
This influx of ions stimulates nerve cells
that send signals to the brain.

• Signals produce visual image from the

brain.
417
Steps involved in the visual cycle are;
• Pigment in photoreceptor in rod cell in retina
absorbs light energy to yield a specific
photochemical product.

• Photochemical product initiates a nerve

impulse.

• Light sensitive form of visual pigment is

regenerated.
418
• In the transmission of light by the brain resulting from
the above changes, it is believed that the bleaching
of rhodopsin increases the permeability of the vesicle
membrane across which there is a normal potential
difference so that Ca2+ flow out of the vesicles thus
triggering the nerve impulse that sends signals to the
brain.

• [There is indication that the same carotenoid

pigments occur in rods and cones but differences in
function have been attributed to different protein
conjugates-rhodopsin in rods and iodopsin in cones.]

419
• Three types of cone cells containing
pigments with different absorption maxima
in the blue (445 nm), green (535nm) and
red (575nm) regions of the spectrum are
present in normal individuals.

• They all contain 11-cis retinal bound to

different proteins that modify the colours
perceived.

420
Metabolic role of vitamin A
• Plays a role in vision as constituent of the visual
pigments (synthesis of visual pigments).

• Promotes growth.

• Confers immunocompetence, i.e. plays a role in

immunity and host’s resistance to infection.

• Helps to maintain the skin and mucous

membranes of the oral cavity and digestive,
respiratory, reproductive and urinary tracts.
421
Deficiency

• Dry thickened skin

• Growth failure
• Poor development of bones and nervous system
• Kidney degeneracy
• Xerophthalmia (inflammation of the conjunctiva)
• Nyctalopia (night blindness)
• Possible death in children.

422
Toxicity (high concentration or heavy doses)
occur because animals are unable to excrete
excess quantities of retinol. Symptoms
include;

• Headache – as a result of elevated pressure

of the spinal fluid around the brain, swelling
around the optic nerve.

• Anorexia, irritability and scaly dermatitis.

• Dizziness, nausea, itching of the skin.

423
• Coarsening and thinning of hair (hair loss).

• Swelling (enlargement) of the liver and

spleen.

• Bone fragility and pain.

• High dose intake during pregnancy

produces foetal abnormalities.

424
VITAMIN D
• It is a complex steroidal alcohol and has several
forms, the active ones being vitamin D2
(ergocalciferol) and vitamin D3 (cholecalciferol).

• The pro-vitamin is called calciferol.

Vitamin D3 (cholecalciferol)
• This is synthesized in the skin of humans and
animals from a precursor 7-dehydrocholestrol
when the individual is exposed to the ultra violet
(UV) component of the sunlight (290-315nm).
425
• 7-dehydrocholesterol is an animal product
formed in the intestines from cholesterol.

• On reaching the skin, it is converted to

cholecalciferol on exposure to direct sunlight.

• It is either stored in the liver or utilized by

the tissues.

426
 enzymatic

HO HO

UV radiation of the skin

cholesterol 7-dehydrocholesterol

H3C
CH3
CH3

H3C
CH2

or
HO
CH2

cholecalciferol or D3 HO cholecalciferol
427
Vitamin D2 (ergocalciferol)
• This is synthesized commercially by UV irradiation of
ergosterol obtained from yeast and other fungi and
plants.

irradiation
CH2

HO

Ergosterol HO

or
ergocalciferol
H3C
CH3
CH3

H3C

CH2

HO
428
Properties
• The D vitamins are white crystalline substances,
soluble in fats and fat solvents.

• They are resistant to heat and oxidation.

• They are not affected by acids or alkali. They are

quite stable if kept stored in the dark in the
absence of oxygen.

Sources; The vitamin is stored in the liver especially

cod liver and some fishes.
Bile salts are required for their absorption. 429
Biochemical function
• D3 itself is inactive but it is hydroxylated in the liver
to form the active form 25-hydroxycholecalciferol.

• This undergoes a further hydroxylation in the

kidney to 1, 25-dihydroxycholecalciferol.

• This conversion is enhanced by parathyroid

hormone and a low level of serum phosphate

430
 cholecalciferol

hydroxylation 25-hydroxylase (liver)

25-hydroxycholecalciferol

hydroxylation 1,-hydroxylase
(kidney)

1,25-dihydroxycholecalciferol
(main physiologically active form)
431
• The dihydroxycholecalciferol directly affects the
absorption of Ca2+ ions from the intestines by
stimulating synthesis of a protein that transports Ca2+
(calcium binding protein).

• It also promotes increased excretion of phosphate by

the kidney and mobilizes calcium ions from the bone
when serum Ca2+ levels fall lower than normal.

• This process is triggered by parathyroid hormone

which acts on the kidney to stimulate release of 1,25-
dihydroxy-cholecalciferol.

• Ca2+ is involved in blood clotting, muscle

contraction and heart function.
432
Functions
• Involved in the mobilization of Ca2+ from the
intestinal tracts for bone building and hence
required for normal growth (required for proper
calcification of bones).

• Involved in calcium and phosphorus metabolism.

• On the whole, the function of vitamin D is to cause

increased absorption, longer retention and better
utilization of calcium and phosphorus in the body.

433
Deficiency
• Causes children to have soft deformed and
poorly calcified bones (rickets).

434
• Causes osteomalacia (soft bones) in adults- a
disease in which there is depletion of bone calcium
already deposited resulting in loss of mechanical
strength. (It occurs in women on poor cereal diet
devoid of milk who keep indoors all-day.)

• The prime cause of deficiency is climatic as the

body can carry out unaided all the stages of
biosynthesis except one, the photo-conversion
stage.
Toxicity
• Excess vitamin D causes bone to become fragile
and easily fracture.
435
VITAMIN E
• It belongs to a class of terpenes called tocopherols. It was
first recognized as a factor in vegetable oil that restores
fertility in rats.

• Early studies showed that rats fed on diet of refined food stuffs
along with cod liver oil and yeast grew at a normal rate and
conceived but did not produce living young as resorption of
foetus occurred.

• The addition of a variety of vegetable foods, especially seed

oils, rendered the animals fertile and resulted in normal birth
of living young.

• The most active in the group is α–tocopherol.

• Vitamin E is stored in muscle and adipose tissue.

436
Structure and properties
The parent structure is tocol with various substitutions at
positions 5, 7 and 8 on the ring system to produce tocopherols

CH3 CH3 CH3

HO CH2 CH CH2 CH CH2 CH(CH3)2

3 3
3

H3C O CH3
CH3 tocopherol
or
CH3

HO
CH3
H3C O
CH3 CH3 CH3
CH3
437
438
• The various tocopherols differ from each other in substituents on carbons 5,
7 and 8.
• These substituents are methyl groups and hydrogen atoms. α-tocopherol
contains 3 methyl groups whereas other tocopherols are short one or two
methyl groups on the aromatic ring

439
Properties
• They are pale yellow oils soluble in fats and
fat solvents.

• They are stable to heat and acids in the

absence of oxygen.

Sources; Seeds and seed oils, vegetable oils,

grains, fruits and animal products,
margarine.
440
Biochemical function
• Vitamin E has high antioxidant activity
preventing the auto oxidation of
polyunsaturated fatty acids.

• It therefore protects unsaturated fatty acids of

biological membranes (in membrane lipids,
e.g. phospholipids and sphingolipids) against
the action of oxygen.

• Being a good reducing agent, it reacts with

oxidizing agents before they can attack other
biomelecules. 441
HC CH CH2 CH CH loss of H+ HC CH CH CH CH A
.

R R R O2 R
AH

HC CH CH CH C .
AOO
OO
R R peroxide
AH
.
A
HC CH CH CH CH
hydroperoxide
R O OH AOOH
R
442
• A free radical has at least one unpaired
electron, which accounts for the high degree of
reactivity.

• Free radicals may play a part in cancer

development and aging.

• The action of tocopherols in the reaction is to

break the free radical chain reaction
mechanism by converting the free radicals into
reactive groups.
. .
AOO +  AOOH + 
. .
A +  AH + 
443
• The tocopherol free radicals (αT.) produced are
not capable of producing other free radicals and
are stabilized by resonance (delocalization).

• They are also prevented from reacting with each

other by steric hindrance.

• They also lack sites to which oxygen can attack to

produce other free radicals.

• Its antioxidant action prevents vitamin A and other

compounds from degradation.
444
445
Deficiency
• Causes infertility in both male and female rats
(not known if it can cause infertility in man).

• Products of autooxidation (brown pigments)

accumulate in fat depot, liver and other
organs where they do not normally occur.

• Degeneracy of the kidney and necrosis of the

liver (death of liver cells).

446
• The normal resistance of polyunsaturated
fatty acids in red blood cell membrane to
oxidizing agents is reduced leading to
spontaneous rupture. (Administration of
vitamin E restores the resistance to
normal).

• Increased susceptibility to haemorrhage.

• Affects muscular system causing

dystrophy and paralysis and if the heart is
affected, myocardial failure may result
leading to death.
447
VITAMIN K
• There are at least 3 forms of the vitamin; K1
(phylloquinone), K2 (farnoquinone) and K3
(menadione).
Structure and properties
O

CH3
CH3 CH3 CH3
CH2 CH C CH2 CH CH2 HC
3 3
2 CH3O
O

K1-phylloquinone (plant source) CH3

O CH3
CH3
CH2 CH C CH2 H
n
O
n=6-9 or 10 depending on the
species
O
Farnoquinone-K2-products of bacterial synthesis
K3-menadione mainly by intestinal flora
448

448
• The natural vitamins are fat soluble and fairly stable to heat and
to reducing agents, however, their activity is abolished by UV
light, strong acids and oxidizing agents, especially in alkaline
solution.

• Menadione is sparingly soluble in water. Synthetic water

soluble forms (like menadione but bears S and P groups) of the
vitamin are commercially available.
OSO2ONa OPO3Na2

OSO2ONa OPO3Na2

K4
449
Source - green leafy vegetables; spinach, cauliflower,
cabbage

Vitamin K2
• A variety of microorganisms including those in the
intestines of higher animals and men synthesize it.

• It may also be obtained from fish meal. K2 from different

sources differs in length of side chain.

• K2 is believed to be the active form of the vitamin which is

required to maintain normal blood clotting time.

450
Biochemical function
• Vitamin K is required for the complex
processes involved in blood clotting.

• It is required for the proper formation of

prothrombin, a blood plasma protein which is
the inactive precursor of thrombin.

• Thrombin is involved in the conversion of

fibrinogen to the insoluble fibrin which holds
blood clots together.
451
452
• Deficiency does not readily occur in rats and other
animals because the vitamin is synthesized by
intestinal bacteria.

• Thus administration of drugs like sulfonamides and

other antibiotics which inhibit the growth of
microorganisms and destroy them can lead to
vitamin K deficiency.

• The vitamin requires bile salts for absorption

therefore any means that eliminates the flow of bile
into the intestines can lead to vitamin K deficiency.

453
• This is why in jaundiced patients the vitamin is
administered due to the absence of bile salts
from the intestinal tract.

• In the neonatal period of life, vitamin K

supplement is needed in the first few days of
life to balance inadequate maternal intake,
reduced permeability of the placenta to the
vitamin, and to the lack of bacterial flora to
synthesize the vitamin in the gut of the new
born.

454