CHAPTER 1

BASIC CONCEPT

INTRODUCTION AND HISTORY

Terpenes constitute a huge and important class of secondary

metabolites. A wide range of active compounds such as perfumes,
cosmetics and food products contain terpenes as part of their
components. Terpenes are a class of natural products consisting of
compounds with the formula (C5H8 )n (where n > 1). The term terpene
was coined in 1866 by the German chemist August Kekule to denote all
hydrocarbons having the empirical formula C10H16, of which camphene
was a member. Kekule coined the term “terpene” in order to reduce
the confusion, as many other hydrocarbons with the same empirical
formula C10H16 were also called “camphene”. The name terpene is the
shortened form “turpentine”, an obsolete spelling of “turpentine”.
Although the term “terpenoids” are used interchangeably with
“terpenes”, terpenoids are modified terpenes that contain additional
functional groups, usually oxygen-containing. Furthermore, terpenes
are produced from terpenoids and many terpenoids are produced from
terpenes. Both terpenes and terpenoids often possess a characteristic,
pleasant odour, which may protect their hosts or attract pollinators.
The inventory of terpenoids and terpenes is estimated at 55,000
chemical entities. In 1939 a Nobel Prize in Chemistry was awarded to
Leopold Ruzicka for his work on polymethylenes and higher terpenes,
including the first chemical synthesis of male sex hormones. The
biogenetic isoprene rule or the C5 rule was described in 1953 by
Leopold Ruzicka and his colleagues.
OCCURRENCE OF TERPENES AND ISOPRENOID COMPOUNDS
Terpenoids, or isoprenoids, are isoprene-based natural products
with fundamental roles in the metabolism of all organisms. Terpenoid
chemical diversity is especially high in plants where many can be
considered secondary metabolites. Such non-essential, specialized
plant terpenoids underlie many ecological interactions between plants
and animals, acting as allelochemicals to attract pollinators, repel
herbivores, or attract herbivore predators. The evolution of terpenoid
secondary metabolism in plants began with the recruitment of genes
from primary metabolism [5] and accelerated due to the proliferation
of cytochrome P450 and terpene synthase gene families in the
genomes of plants. Terpenoid chemical diversity partly reflects a
natural history marked by herbivory stress and other selective
pressures imposed by animals, resulting in a broad array of
functionalized terpenoids in the plant kingdom pre-selected for their
potent biological activities towards animals. This selective process may
have been facilitated by the general similarity of protein folds and
motifs between plant and animal proteins, resulting in plant secondary
metabolites with natural affinity for animal proteins by virtue of having
been produced by plant enzymes composed of the same amino acids.
Thus, pre-selection and emergence of plant secondary metabolites with
biological activity towards animals may have both an evolutionary and
a biochemical basis.
As a consequence, many plant terpenoids have found fortuitous
uses in medicine, and the terpenoid family of natural products has been
a valuable source of medical discoverie. Hundreds more allegedly
possess medicinal properties, but the testing process is painstaking and
resource intensive. The true number of plant terpenoids in nature
which could potentially be screened for therapeutic applications is
unknown but is likely >105, including >12,000 from the diterpenoid
group alone. While this number is small compared to modern
combinatorial methods, the lead compound discovery rate may be
significantly higher for plant natural products due to the
aforementioned pre-selection effects. But owing to their high degree of
chemical functionalization and metabolic specialization, many are
produced in small amounts, only in response to elicitation, or
accumulate exclusively in specialized tissues, necessitating microbial
production or major advances by plant breeding and genetic
improvement to obtain sufficient quantities to investigate clinical
potential. 

CLASSIFICATION OF TERPENES
A) Based on the number of isoprene units
i. Hemiterpenes (1 isoprene unit)
ii. Monoterpenes (2 isoprene units)
iii. Sesquiterpenes (3 isoprene units)
iv. Triterpenes (6 isoprene units)
v. Tetraterpenes (8 isoprene units)
vi. Polyterpenes ( > 8 isoprene units)
B) Based on the nature of carbon chain
i. Acyclic terpenes
ii. Monocyclic terpenes
iii. Bicyclic terpenes
iv. Tricyclic terpenes
v. Tetracyclic terpenes
vi. Pentacyclic terpenes
ISOPRENE (2-METHYL BUT-1, 3-DIENE)
 CHAPTER 2
BIOSYNTHESIS OF TERPENES
The C5 isoprene units are provided in the form of dimethylallyl
pyrophosphate (DMAPP) and isopentenyl pyrophosphate (IPP). DMAPP
and IPP are structural isomers to each other. This pair of building blocks
are produced by two distinct metabolic pathways: the mevalonate
(MVA) pathway and the non-mevalonate (MEP) pathway. These two
pathways are mutually exclusive in most organisms, except for some
bacteria and land plants. In general, most archaea and eukaryotes use
the MVA pathway, while bacteria mostly have the MEP pathway. IPP
and DMAPP are final products of both MVA and MEP pathways and the
relative abundance of these two isoprene units is enzymatically
regulated in host organisms. Plants may however utilize either of the
two pathways in the synthesis of terpenes.
The Mevalonate Pathway
This pathway conjugates three molecules of acetyl CoA.
Hydroxymethylglutarate, an intermediate is reduced using NADPH and
HMG-CoA reductase to give Mevalonate (hence the name “the
mevalonate pathway”). Mevalonate is then Phosphorylated at the
carbon 5 position to give mevalonate pyrophosphate (5-
pyrophosphomevalonate).
Isoprenoid synthesis then proceeds by the condensation of isopentenyl
pyrophosphate (gotten from the decarboxylation and dehydration of
mevalonate pyrophosphate) with the isomeric dimethylallyl
pyrophosphate to yield geranyl pyrophosphate. Further C5 units are
added by the addition of more isopentenyl pyrophosphate. From
geranyl and farnesyl pyrophosphate various structures can be built up.
It is however important to note that only the (R) form of mevalonic acid
gives rise to the terpenoids, the (S) is biologically inactive
The Non-mevalonate pathway
The non-mevalonate pathway or the 2-C-methyl-D-erythritol 4-
phosphate (MEP) pathway starts with pyruvate and glyceraldehyde 3-
phosphate (G3P) as the carbon source.
C5 IPP and C5 DMAPP are the end-products in either pathway, and are
the precursors of terpenoids with various carbon numbers (typically
C5 to C40), side chains of (bacterio)chlorophylls, hemes and quinones.
Synthesis of all higher terpenoids proceeds via formation of geranyl
pyrophosphate (GPP), farnesyl pyrophosphate (FPP),
and geranylgeranyl pyrophosphate (GGPP).
 CHAPTER 3
THE PHYSICOCHEMICAL PROPERTIES OF TERPENES

Most of the terpenoids are colourless, fragrant liquids which are lighter
than water and volatile with steam. A few of them are solids e.g.
camphor. All are soluble in organic solvent and usually insoluble in
water. Most of them are optically active.

 They are open-chain or cyclic unsaturated compounds having one

or more double bonds. Consequently, they undergo an addition
reaction with hydrogen, halogen, acids, etc. A number of
additional products have antiseptic properties.
 They undergo polymerization and dehydrogenation.
 They are easily oxidized nearly by all the oxidizing agents. On
thermal decomposition, most of the terpenoids yield isoprene as
one of the products
 They are highly flammable being hydrocarbons and possess a low
specific gravity (I.e. they float on water)
 They are usually local irritants and can cause gastrointestinal
disturbances if ingested
 Terpenoids have similar properties but tend to be more polar and
less volatile than their terpene analogues..
 CHAPTER 4
CHEMISTRY AND CHEMICAL STRUCTURES

Chemically, terpenes are grouped together because of their distinctive

carbon skeleton. It consists of a basic five-carbon isoprene unit (2-
methyl-1,3-butadiene). Terpenes generally are composed of two, three,
four, or six isoprene units. These are called monoterpenes,
sesquiterpenes, diterpenes, and triterpenes, respectively.

Terpenes may contain a variety of functional groups. Many important

terpenes contain hydroxyl groups, making them terpene alcohols.
Other terpenes are ketones. Terpene oxides are terpenes having an
oxygen-containing ring structure, as well as the basic isoprenoid
structure. As such, they contain a cyclic ether (C-O-C) bond.
Biogenetic Isoprene rule: the very idea that terpenes are essentially
built up of several isoprene units is commonly termed as Biogenetic
isoprene rule.
Meroterpenoids: it has been observed that a good number of other
natural products do exist which essentially belong to mixed
biosynthesis origin and are mostly made up from isoprene as well as
non-isoprenoid entities (e.g: cannabinol, quinine, vitamin-E)

Ergitomaine (an ergot alkaloid with one isoprene unit)

The International Union for Pure and Applied Chemistry recommends a
systematic mode of nomenclature of terpenoids, however, the names
suggested are lengthy and quite cumbersome. Therefore, the old and
trivial names most terpenoids are used more frequently, even as of
today.
Examples:
Trivial Name IUPAC Name
Geraniol 3, 7 – Dimethyl-2, 6-octadiene-1-ol
Limonene 1-Methyl-4-(1-methylethylnyl)-cyclohexane

Monoterpenoids

In general, monoterpenoids represent a structurally diverse class of

compounds maybe categorized into nearly 35 varying structural
analogues, However, the most commonly occurring structural
variations are of the following types, namely:
1) Acyclic:

Myrcene (an acyclic monoterpene)

 2) Monocyclic:

Menthol (a monocyclic monoterpenoid)

3) Bicyclic:

examples of bicyclic monoterpenes

Sesquiterpenoids

The sesquiterpenoids are found to be extensively distributed in nature

and by all means represent the most abundantly prevailing class of
terpenoids.
They may be classified into four major categories namely: acyclic,
monocyclic and tricyclic sesquiterpenoids.
Other classes of Sesquiterpenoids include:
Sesquiterpenoid Lactones: these constitute a class of compounds
essentially bearing characteristic features as an alpha- methylene
gamma-lactone system, alpha, beta-unsaturated carbonyls and
epoxides and other chemically distinct features, are collectively called
sesquiterpenoid lactones.
An example of a pharmaceutically useful natural product that possesses
the sesquiterpenoid lactone functions include: Artemisinin

Artemisinin

Diterpenoids
They represent a broad class of non-volatile C20 compounds that have
been essentially obtained from geranyl pyrophosphate.
Diterpenoids possess the following chemical features:
1) Most of them are carboxylic compounds having upto five aromatic
rings
2) Certain members of this class are acyclic compounds
3) They mostly occur as hydrocarbons or highly oxygenated
compounds based on their degree of oxidation,
4) They are invariably isolated as optically active solids

Triterpenoids
They are generally obtained by biogenesis of six isoprene units,
they are found to share the common acyclic precursor squalene
(C30).
The triterpenoids may be categorized into two major groups,
namely: the tetracyclic and the pentacyclic compounds.
Interestingly, both groups may easily combine with sugar moieties
at C-3 position to yield the corresponding glycosides.
Nevertheless, the triterpenoids are invariably associated with
natural latex, resins, or cuticle of plants.

Tetrapenoids
A plethora of natures yellow, orange, red and purple colours are
mostly by virtue of the presence of carotenoids. They essentially
consist of Tetraterpenoids. A typical example being vitamin A

Chemistry of some common terpenoids

Thymol:
 The phenolic OH moiety present in thymol enables it to form
salts of acetate and carbonate easily which are applies in its use
as an antiseptic and anthelmintuc respectively
It may be synthesized by the treatment of menthone first
with bromine and then with quinolone.
Thymol when dissolved in NaOH solution and treated with
an I2-KI solution forms thymol iodide that finds it use as an anti-
infective and antifungal agent.it is a monocyclic monoterpene

Ginkgolide-B:

It is the most active member of the family with significant

therapeutic efficacy in the treatment of severe sepsis, whereas the
 corresponding A and C analogues are devoid of such abilities. It is
a Diterpenoid

Camphor:

camphor is a bicyclic terpenoid

ketone, specifically a monoterpenoid, as it contains two isoprene units.
In the presence of platinum black it undergoes hydrogenation at
ambient temperature giving rise to isoborneal as the major product and
traces of borneal. On prolonged hydrogenation it yields camphene.
 CHAPTER 5
GENERAL AND SPECIFIC CHEMICAL TESTS FOR TERPENES/TERPENOIDS

Chemical Test For Terpenoid :- 

1. Liberman-Burchard Test :-  Drug is mixed with acetic anhydride.

To this mixture conc. Sulfuric acid is added. There forms two
layers with browning at junction. Upper Layer with green colour
represent steroids whereas lower layer represent Terpenoid red
colour.
2. Salwoski Test :- Drug is mixed with con. Sulfuric acid. Upper layer
is of steroids which are red in colour and lower layer yellow colour
layer represent tri terpenoid.
3. Sulphur Powder Test :- If sulfur is added to the mixture of drug,
sulfur sinks down the mixture.
4. Trichloroacetic acid Test :- Coloured PPT
5. Tetranitro Methane Test :- Yellow PPT
6. Zimmer mann Test:-  Meta dinitro benzene solutions was added
to the alcoholic solution of drug containing alkali, on heating →
Violet Colour (Keto-Steroid)

Specific Test for Terpenoid :-

1. Bitter Orange Peel :- Solutions of recently express orange oil in

dehydrated alcohol is neutral to moistened litmus paper.
2. Camphor Oil :-  A drop of solutions of Vanillin (1:100) and H2SO4
when added to powdered natural camphor, produce immediately
a yellow colour changing to red, violet and finally blue Colour.
3. Chenopodium Oil :-  Heat 1 ml chinopodium oil in a test tube with
a piece of porcelain. A deep golden yellow colour is produced.
4. Cinnamon Oil :- On addition of a drop of FeCl3 solution to a drop
of volatile oil produces a pale green colour. With ferric chloride,
cinnamic aldehyde give brown colour and Eugenol give blue
colour result the formation of pale green colour.
5. Eucalyptus Oil :- Mix 2.5 ml of Eucalyptus oil is with 5 ml of
purified petroleum benzene add 5 ml of solutions of sodium
nitrite. Then add 5 ml of glacial acetic acid crystal of phellandrene
nitrite do not form in the mixture within 10 minutes.
6. Rasna :-  Shake About 1g finely powdered rasna with 60% alcohol
for 10 minutes and filter put a drop on filter paper and dry it.
Examine the same under UV light, a bright blusih white
florescence is observed.
 VOLATILE OILS (or ESSENTIAL OILS)
Volatile oils or ethereal oils are the odorous and volatile products of
various plant and animal species. They differ from the fixed oils in that:
1) They mostly consists of terpenoids
2) They leave no spot or grease spots on filter paper
3) Saponification tests are not applicable to them
4) They possess distinctly marked and specific fragrance
5) The are also easily oxidized on exposure to air and light and
undergo resinification

They are prepared using the following methods:

1) Direct steam distillation
2) Hydro-distillation
3) Extraction (solvent or super critical fluid extraction)
4) Fractional distillation
5) Enzymatic hydrolysis

Common essential oils that are also terpenes include: camphor,

eucalyptus oil, oil of turpentine, menthol, limonene, citral, pinene,
carvone, linalool. They are mostly derived from Monoterpenes,
however some volatile oils also possess sesquiterpene features
 CHAPTER 6
MEDICINAL AND PHARMACEUTICAL USES OF TERPENES/ TERPENOIDS
AND VOLATILE OILS
Apart from the flavor that gives to food, essential oil contain
antimicrobial properties. Thyme is one of plants that synthesize
terpene alcohols and phenols which contain powerful antibacterial and
antifungal properties.
Medicinal Properties of terpenes from different sources
Terpene Medicinal properties

Contains the active ingredient to treat cutaneous

Tea tree
infections

Possesses powerful antibacterial and antifungal

Thyme
properties

Possesses psychoactive properties and used against

Cannabis
many infectious diseases

Spanish
Enhances memory and is used in anti-dementia drugs
sage

Citrus fruits Drugs against pediculosis

Citral Antibacterial and antifungal effects

Lemongrass Insect repellent

MENTHOL
 It is used in various mouthwashes and toothpastes 
  It is used as a flavoring agent in some pharmaceutical
formulations such as cough syrups and lozenges
 It is used on the mucous membranes or on the skin to serve as a
counterirritant, antiseptic and as a mild stimulant
 It is combined with other allied substances such as camphor and
eucalyptus oil as an expectorant and nasal decongestant
 Menthol also aids in the dilation of blood vessels and is employed
in combinations with other drugs in the relief of rheumatoid pains
ARTEMISININ
 The drug and its derivatives are used as fast acting blood
schizontocides in the control and management of malarial
fever caused by plasmodium vivax
 It is beneficial in the treatment of cerebral malaria and
chloroquine resistant malaria.
 It has been listed as a potential COVID-19 drug due to its
high anti-SARS-CoV-2 activity

TAXOL (PALCLITAXEL)
 It is used to treat breast cancer, ovarian cancer and lung cancer
 It is also used to treat AIDS-related Kaposi’s sarcoma

structure of taxol
 REFERENCES

 Pharmacognosy and Pharmacobiotechnology by Ashutosh Kar

 Trease and Evans Pharmacology 16th edition by William C. Evans
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and function of plant isoprenoids.Prog.LipidRes.2005,44,357–429.
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 12.Lange, B.M.Online resources for gene discovery and
biochemical research with aromatic and medicinal
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