3.

Bicyclic monoterpene hydrocarbons

The bicyclic monoterpenes may be divided into three class according to the size of
the second ring. The first being a six membered ring in each class. It is important
to note that the two rings do not exist in one plane, but are almost perpendicular to
each other.

Class l. (6 + 3 membered ring)

Thujane Carane
Class ll: (6 + 4 membered ring)

Pinane
Class Ill: (6 + 5 membered ring)

Camphane Iso-Camphane Fenchane Isobornylene

Class I: (6 + 3 membered ring)

Terpenes of the thujane structure:

36
 Sabinene (Savin oil) α- Thujene (Juniper oil)

Terpenes of the carane structure

Source: Turpentine oil.

Uses: As precursors of….sylvestrene and carvestrene.

Class II: (6 + 4 membered ring)

Terpenes of the pinane structure:

β-pinene (Nopinene) α- Pinene

α- Pinene is very widely distributed in nature, being found in most volatile oils of
the Coniferae. It is the main constituent of oil of turpentine. The turpentine oil is
separated from the resin (colophony) by steam distillation.

Pure α-pinene is obtained from the above fraction of turpentine oil by repeated
fractional distillation, followed by conversion to the crystalline nitrosochloride,
from which α-pinene, is liberated by treatment with aniline.

α-pinene is used as starting material for different synthetically prepared

compounds:
1- Preparation of borneol and camphor:

37
α-pinene identified by preparation of a mono nitrosochloride m.p. 115°C, and
mono hydrochloride m.p 132°C. (due to the presence on ethylenic linkage).

α-pinene is of great importance commercially in the synthesis of camphor and

related terpenes.
β-pinene, which is the isomeric with α-pinene, accompanies the latter in most
sources of the hydrocarbon. The two compounds may be separated by careful
fractionation. β-pinene distilIs at 162-163°C.

Class III: ( 6 + 5 membered ring)

Terpenes of the camphane structure:

Camphene

Source: Nutmeg oil.

Camphene is the only solid crystalline terpene hydrocarbon m.p. 51-52°C.

38
Terpenes or the fenchane structure

Fanchene

Source: French turpentine oil.

Sequiterpene hydrocarbons
The sesquiterpenes, in general, form the higher boiling fraction of the volatile
oils. The sesquiterpene structure is built up of three isoprene units.

The sesquiterpenes are classified into four groups according to the number of
ring present in the structure. If we use the isoprene rule, then when three
isoprene units are linked (head to tail) to form acyclic sesquiterpene
hydrocarbon, the later contain four double bond. Each isoprene unit contains
two double bonds, but one disappears for each pair that is connected

When this open-chain compound is converted into a monocyclic structure another

double bond is utilized in the process, and so monocyclic sesquiterpene
hydrocarbons contain three double bond. In a similler mannar it will be found that
a bicyclic structure contains two double bonds, and a tricyclic structure contains
one.

1. Acyclic ………….4 double bond……….... no ring

2. Monocyclic……..3 " …………one ring
3. Bicyclic………….2 " ………… two rings
4- Tricyclic…………1 " ………… three rings

Most of these sesquiterpene hydrocarbons are faint odour, more viscous. than
39
the terpenes, sparingly soluble in alcohol. They are readily resininfied. Being
unsaturated hydrocarbons, they form halogen compounds, hydrogen halides.
nitrosochloride, nitrosites and nitrosates. Some of them are crystalline, and
therefore helpful in the Identification of the parent sesquiterpene.

1-Acyclic sesquiterpene
Sesquicitronelline

α-Farnesene ß- Farnesene
(Oil of Citronalla) (Oil of Hops)

2. Monocyclic sesquiterpene
Zingiberene

Source: Oil of ginger

3. Bicyclic sesquiterpene
A. Derivatives of naphthalene
Cadinene

Source: Oil of cade.

40
 B. Derivatives of azulene

Azulene ( C10 H 8 )

Azulene is a non-benzenoid aromatic compound. The natural azulenes are

coloured which vary in colour from blue to violet or even green and are of various
structure, They occur in several volatile oil e.g. cubebs, valerian. galbanum and
chamomile or may occur in the form of colourless sesquiterpene precursors,
which by physical or chemical action can be transformed into coloured
azulenes. Most azulenes are well crystallized substances m.p.40-100°C.

Isolation and identification of azulenes:

Azulenes are isolated in the form of derivatives or adducts which are also used
for their identification.
1. Azulenes form addition compounds with strong mineral acids as phosphoric
or sulphuric acids. These addition compounds can be easily decomposed with
water to regenerate the azulenes.
2. Azulenes form ferrocyanate with ferrocyanic acid, which can be decomposed
with water to regenerate the azulene.
3. Azulene form picrate, trintrobenzene derivatives with sharp melting points.
From these derivatives azulenes can be regenerated in pure form with sodium
hydroxide.

Guaiazulene

41
Blue needles, m.p. 31°C
Source: - Geranium oil.
- Dehydrogenation of V. O. of guaiacum.

Vitavazulene

Violet needles, m.p. 33°C.

Source: Heating vetiver oil with
sulphur.
Chamazulene

Chamazulene .....chamomile oil (about 15%, blue colored).

C- Miscellaneous sesquiterpenes
Caryophyllene

Source: Clove oil.

Humulene

42
 Source: Oil of clove & oil of hops (The isomer of caryophylline).

4- Tricyclic sesquiterpene
α-Santalene

Source: Oil of sandalwood, (Santalum album).

Aromatic hydrocarbons
P-Cymene

Surce: Oil of lemon.

Terpeneless and sesquiterpeneless oils
Certain of the volatile oils are available commercially in a form known as
terpeneless oils and sesquiterpeneless oils. In the first type, only the terpenes
have been removed; in the latter, both the terpenes and sesquiterpenes have
been eleminiated. Because the terpenes and sesquiterpenes oxidize and resinify
easily under the influence of air and light or under improper storage condition
which means spoilage of odour and flavour, and lowering of the solubility in
alcohol.
This procedure of elimination involves distillation under high vacuum, washing
with dilute alcohol (in which the oxygenated constituents are soluble, but not the

43
hydrocarbons).
Terpeneless and sesquiterpeneless oils are much more expensive than the
original oils, but they possess certain advantages which render very useful. They
have much higher flavouring and odoriferous properties than ordinary oils. in
some instances up to 35 or 40 times as much.

Oxygenated terpene compounds

The oxygenation of the terpene hydrocarbons would naturally account for the
presence of (1) alcohols, (2) aldehydes. (3) ketones, (4) phenols, (5) phenolic
ethers, (6) esters and (7) oxide. Sine these oxygenated compounds are responsible
for the characteristic odour, taste, and therapeutic properties of the volatile oils.

1- Alcohols
The separation of terpene alcohols from the volatile oils is accomplished by a
variety of methods such as:
1- Fractional distillation. 2. Counter current extraction.
3. Preparation of derivatives. 4- Gas chromatography.

Most terpene alcohols are conveniently separated from volatile oils via their
crystalline additive products, which is very useful for the identification and
purification of the alcohols.
The general methods which used for separation of terpene alcohols are:

1- Calcium chloride addition products

Many primary alcohols, among them geraniol, cinnamyl alcohol, benzyl alcohol
and butyl alcohol form complex crystalline salts when shaken with powered
anhydrous calcium chloride in absolute ether or in benzene; or some times in
absence of solvent. The original alcohol is easily, regenerated by the addition of
water, geraniol addition product is the most important one, since it affords a
means of separating this alcohol from similar primary alcohol nerol and citronellol),

44
2. Formation of Esters
As a general rule, primary, secondary, and tertiary alcohols differ greatly in respect
to the speed with which they react to form esters. With acid anhydrides, primary
alcohols react more readily than secondary alcohols, and tertiary alcohols are not
appreciably affected, Certain esters are useful for the isolation, purfifcation as.
well as identification of alcohols such as Borates (Boric acid esters), phthalates
(phthalic acid esters), and Benzoates (Benzoic acid esters).

3- Dehydration products
Most tertiary alcohols are unstable and split off water, when reacted upon by
reagents generally employed in the preparation of alcohol derivatives.
Dehydration products (usually hydrocarbons) will result, which may be useful for
the identification of an alcohol, if the dehydration product can readily be identified.
Dehydration methods are generally based upon catalytic decomposition of tertiary
alcohols with gentle heating.

Classification of terpene alcohols

Alcohols occurring in volatile oils may be classified into (A) acylic alcohols, (B)
alicyclic alcohols, (C) sesquiterpene alcohols and (D) aromatic alcohols.

(A) Acyclic terpene alcohols

CitroneIlol

Source: Citonella oil.

Separation:
 Fractional distillation.

45
  Formation of esters (acid phthalate).
Identification :
 Formation of esters (phthalate), citronellyl acid phthalate..yield crystalline
silver salt m. p. 125-l26°C.
 Citronellol….(oxidation with chromic acid)….citronellal (aldehyde)……
..(addition of specific carbonyl reagenrt, semicarbazide)…..Crystalline
adduct (semicarbazone)……………Identified by its melting point.

Uses: used in perfume industries.

Geraniol and Nerol

Geraniol (trans-form) Nerol (cis-form)

Source: Geraniol….. Oil of geranium.
Nerol………Oil of neroli.
Separation :
 Fractional distillation.
 Formation of esters (acid phthalate).
Identification :
 Preparation of phenyl urethane derivatives (crystalline compounds), can be
identified by its m.p.
 By oxidation….. Geraniol gives ….Citral a , Nerol gives….Citral b.
Uses: Geraniol and nerol are used extensively in perfumery, especially in the
preparation of systhetic rose scents.

Linalol (Linalool)

46
It is 3 ry alcohol.
Linalol….. Oxidation………………..Linalal (aldehyde = citral).
Source: Oil of linaloe.
Uses: Linalol is used extensively in perfumery, especially in the form of its esters.

(B) Alicyclic terpenene alcohols

The alicyclic terpene alcohols are classified into three groups, (1) monocyclic, (2)
bicyclic and (3) sesquiterpene alcohols.

1- Monocyclic terpene Alcohols

Menthol

Source : Japanese peppermint oil (Mentha arvensis), 75-90% menthol.

Separation from the oil :
Menthol is usually prepared from The oil is cooled to + 15ºC in a freezing mixture and
the deposited menthol crystals are separated by centrifugation of the oil. The
remaining oil is then cooled to + 5°C and the crystals again removed by centrifugation.
The process is repeated a third time at – l0°C. Starting from a natural oil containing
about 85% I-menthol, the residual oil, after the third treatment, will still contain 40-50%
natural l-menthol and some menthone (its corresponding ketone). This ketone can he
removed from the mixture by conversion into its oxime. The oxime is extracted from the
etherial solution through shaking with dilute sulphuric acid. The I-menthol will be
obtained on cooling in solid crystalline form, m.p. 42- 43°C.

47
The solid portion separated from the volatile oil by cooling is called "stearoptene", and
the liquid portion is called "eleoptene".

Menthol is also prepared synthetically (dl-). Synthetic racemic menthol is produced by

hydrogenation of thymol.

Menthol occurs in colourless, hexagonal crystals, usually needlelike, or in fused

masses, or a crystalline powder, and has a pleasant, peppermint like-odour.

Reactions of menthol:
Menthol is a saturated, secondary alcohol, yielding on oxidation the cooresponding
ketone methone.. On dehydration it yields a menthane, which on dehydrogenation
affords p-cymene.

Menthol is identified by the following method:

1. Determination of the melting point (42-43°C).
2. Preparation of derivatives, such as p-nitrobenzoate, phosphoric
acid complex, acid phthalate and menthoxy acetate.

Color tests :

a. Few crystals of menthol are dissolved in few drops of conc H 2 So4 and few drops
of vanilIin/sulphuric acid reagent are added, an orange yellow colour is developed
which changes to violet on the addition of few drops of water.
b- Few crystals of menthol are dissolved in glacial acetic acid then 3 drops of con.

H 2 So4 and one drop of HNO3 are added, no green colour is produced (c.f. thymol

which gives a green colour).

Uses:
Menthol is a local antipruritic. It has been used on the skin or mucous membranes as
a counterirritant, an antiseptic, and a stimulant.
Menthol is chiefly used to relieve symptoms of bronchitis, sinusitis and similar

48
conditions.
α-Terpineol

It is a 3 ry alcohol.
Source: Oil of camphor.
Uses: α-Terpineol, which has a delicate odour of lilac, is used extensively in the
manufacture of perfumes and cosmetics. It is prepared on an industrial scale from
various pine oils.

2. Bicyclic Monotorpene Alcohols

Borneol

Source: Borneol occurs in oil of Dryobalanops species,

isoborneol do not exist naturally, it is prepared
synthetically.
Uses: Borneol is used for scenting all kind of technical preparations, room sprays,
inhalants, soaps. Some bornyl esters are used in pharmaceutical preparations.

C. Sesquiterpene alcohols
α-Santalol

49
Source: East Indian sandal wood oil.
Uses: α-santalol is used medicinally in the treatment of gonorrhea.

D. Aromatic alcohols
Benzyl alcohol

Source: As esters of benzoic acid and cinnamic acid (benzyl benzoate and benzyl
cinnamate) in balsam peru, balsam tolu and storax.
Uses: Benzyl alcohol is used in perfumes and also serves as an important fixative and
a general diluent of perfume mixture.

Phenylethyl Alcohol

Source: Oils of geranium.

Users: Used in perfumes and cosemetics.

Cinnamyl Alcohol

50
Source: As ester in several volatile oils gums, balsam and resin.
Uses: Cinnamyl alcohol is used and its esters in the perfumes, cosmetics, soap
industries, it is serve as excellent fixative.

Determination of alcohol content in volatile oils

The total alcohols present in any given oil are determined by transforming the free
aIcohols into the corresponding acetate by boiling the oil with acetic anhydride in an
acetylization flask and then determining the saponification value of the acetylized
product.
When the alcohol occurs partly free and partly combined in the form of an ester, a
correction factor must be employed.

Tertiary alcohols are not quantitatively converted to the acetate by this process of
acetylation (e.g. linaleol and α -terpineol).

Determination of primary alcohols:

The primary alcohols present in any given oil are determined by transforming the free
alcohols into the corresponding acid phthalate esters by boiling the oil with phthalic
anhydride. This reaction takes place readily at a temperature of about 100ºC in the case
of primary alcohols, for secondary alcohols, the time required for reflux is greatly
increased; for tertiary alcohol, no appreciable reaction occurs.

Determination of tertiary alcohols:

Most tertiary terpene alcohols are decomposed or dehydrated when treated with
acetic anhydride and a special techniques are required.

1. Formylation method:
(using acetoformic acid) This method is used specially for the determination of linalol
and α- terpineol.

51
2- Use of diluent in acetylation method:
This method of acetylation makes use of a diluent (oil of turpentine, xylene) in order to
decrease the dehydrating effect of acetic anhydride. The time of acetylation,
however, must be prolonged.

3- Dehydration method:
This method is based upon the catalytic decomposition of tertiary alcohol and the
splitting off of water. The amount of water obtained is determined, from which the
percentage of tertiary alcohol may be calculated.

4- Acetyl chloride-dimethyl aniline method:

This method is used mainly in the case of linalol and linalol containing oils. The
alcohol is treated with dimethyl aniline, acetyl chloride and acetic anhydride and the
mixture is heated at 40ºC for 3 hrs.

Determination of citronellol by formylation:

Most terpene alcohols are dehydrated by strong formic acid. giving rise to
nonsaponifiable terpenes. An exception is citronellol which is converted almost
quantitively to corresponding formate. This result is convenient and satisfactory method
for the determination of citronellol in the presence of geraniol and linalol.

2- Esters
The most common esters in many volatile oils are the acetates and formates. Some
volatile oils consist almost entirely of esters; oil of wintergreen, for example, consist
almost entirely of methyl salicylate and contain up to 99% of this ester.

Esters are isolated by the following methods:

1. By cooling of the oil for esters, which solidify at low temperature.
2. By fractional distillation under vacuum.

Esters could be identified by the following methods:

52
1. Saponification of the ester into alcohol and acid and converting the acid into a
characteristic derivatives.
2. By reaction with amines which has proved very valuable identification.
RCOOŔ + C6H5-CH2-NH2 RCONHCH2- C6H5 + ŔOH
Ester Benzyl amine N- Substituted benzyl amine

A) Esters of aliphatic acids

Geranyl acetate
Source: Oil of citronella.

Linalyl acetate
Source: Bergamot oil.

Terpinyl acetate
Source: Cajuput oil.
Bornyl acetate
Source: Pine oil.

Menthyl acetate
Source: Pippermint oil.

B) Esters of aromatic acids

Benzyle benzoate
(Benzoic acid benzyl ester)

Source: Oil derived from balsam peru.

Uses: Used as a fixative in perfume compositions and medicinally for treatment of
scabies.
53
 Methyl cinnamate
(Cinnamic acid methyl ester)
Source: Ocimum oil.

Cinnamyl cinnamate
(Cinnamic acid cinnamyl ester)
Source: It occurs in balsam peru.

Methyl salicylate
(Salicylic acid methyl ester)
Source: Oil of winter green.

Methyl salicylate does not occur as such, but in the form glycoside (gaultherin) which
must be hydrolysed by the enzyme of action (gaulthrase) in order to liberate the ester.

Uses: It is used as local irritant and antirheumatic.

Determination of esters:
When the esters are refluxed with alcoholic potassium hydroxide, they are
saponified and yield the free alcohol and the potassium salt corresponding to the
acid component of the ester.

The ester value of oils that contain appreciable amount of aldehydes or phenols
cannot be estimated accurately by saponification with an alkali, since variable
amounts of the alkalies are consumed by reaction with the aldehydes and
54
phenols, or their decomposition products.

3- Aldehydes

The aldehydes occurring in volatile oils are mainly aliphatic or aromatic. Relative to
the aliphatic, the aromatic aldehydes play a much important role in volatile oils. with
the exception of citral and citronellaI, aliphatic aldehyde do not play an important
role. The aromatic aldehyde include benzaldehyde (bitter almond oil), cinnamic
aldehyde (cassia oil), vanillin (vanilla, benzoin, tolu and peru balsams), and cuminic
aldehyde.
In general, these aldehydes gives all ordinary reactions of the aliphatic aldehydes
as: formation of cyanohydrin, bisulphite complex, oxidation to acids (by reduction
of amm AgNO3 & cooper salts), reduction to alcohols and polymerization.

These reaction may play an important role in the isolation and identification of these
aldehydes. Moreover, these aldehydes readily yield many crystalline derivatives,
typical of the carbonyl group as: Oxime ( with hydroxylamine HCl.),
semicarbazones' (with semicarbazide), and substituted hydrazones.

Acyclic terpene aldehydes

Citronellal (Rhodinal)

55
 Acyclic terpene aldehydes
Citronellal (Rhodinal)

Method of isolation :
1. The oil is shaken with sodium bisulphite solution.
2. The crystalline bisulphite addition product is purified by washing with alcohol
and ether.
3. The citronellal is regenerated by decomposition the sodium bisulphite compound
with sodium carbonate followed by distillation under vacuum.

Citronellal is obtained by mild catalytic hydrogenation of citral. Reduction of

citronellal with sodium amalgum yields the terpene alcohol citronellol.

Uses: Citronellal is an important constituent of many artificial perfumes and

flavouring matters.

56
 Citral

(trans) (Cis)
Natural citral is mainly citral-a.
Source: Lemon grass oil.
Uses: Citral has a powerful odour and taste of lemons. It is used in the compounding
of synthetic lemon flavours, and in perfumery.

Aromatic aldehydes
Benzaldehyde

Source: Bitter almond oil.

Uses: Used as flavouring agent in food stuffs.

This aromatic aldehyde probably does not occur as such in plants. but in the form of
glycoside amygdalin in bitter almond kernels and also in other rosaceous kernels,
as peach and apricot. Benza]dehyde is produced as a result of the reaction of the
enzyme emulsin on the glycoside amygdalin in an aqueous mixture of ground
almond.

57
In bitter almond oil, benzaldehyde is therefore often accompanied by small quantities
of hydrocyanic acid.

Method of isolation:
Benzaldehyde is prepared from the volatile oils of bitter almond, peach and apricot
kernels, These volatile oils contain benzaldehyde, benzaldehyde cyanohydrin and the
toxic HCN, which should be removed first. The HCN is removed by shaking the oil with
milk of lime, and ferrous sulphate and allowing the mixture to stand for a long time,
then distilling the benzaldehyde which liberated with steam.
By this treatment the HCN is converted into calcium ferrocyanide which is not
volatile.

Salicylaldehyde

Source: Cassia oil.

Source: Oil of cumin.

58
Uses: Cuminaldehyde is used for preparation of synthetic cumin oil.

Source: Cassia leaf and bark oil and of cinnamon bark oil.
Uses: Cinnamic aldehyde is used as a flavouring agent.

Vanillin

Source: Vanilla pods.

Uses: Vanillin is used mainly as flavouring agent.

Methods of preparation:
1. From vanilla pods by extracting the powder with ether, evaporating the ether and
crystallization from alcohol.

59
2. Chemically vanillin is prepared from eugenol, the main constituent of oil of cloves.

3. Vanillin is prepared commercially in large amount from guaiacol (catechol methyl

ether) by the Reimer-Tiemann's reaction).

4. Preparation from the glycoside coniferin by oxidation.

5. Commercial method by oxidation of lignin, which is a by-product in manufacture of

paper pulp; lignin is a complex polymeric structural material of woody plants and
contains as a structural unit, the fragment.

60
 Upon oxidation of lignin, vanillin is formed in a high yield to make the process
economically feasible.
4- Ketones
Ketones occurring in volatile oils are either, aliphatic, cyclic or aromatic.
The aliphatic ketones are not of common occurrence in volatile oils. The lowest
members of this group (acetone and diacetyl) originate mostly from decomposition of
the more complex compounds during steam distillation.

The cyclic terpene ketones are of common occurrence and are classified into
monocyclic and bicyclic.
The aromatic ketones are more rare in volatile oils than aliphatic ketone.

A- Monocyclic terpene ketones

Menthone

Source: Pippermint oil.

Uses: Menthone is used extensively in the manufacture of flavouring materials.

Piperitone

61
Source: Eucalyptus oils.
Uses: Piperitone is used as staning material for the preparation of synthetic menthol
and thymol.

Carvone

Source: Oil of caraway.

Uses: Carvone has the odour of caraway fruit, and is used for flavouring of many
kinds of food products and beverages. It is also used in mouth preparations, gargles
and tooth pastes.

Diosphenol

Source: Volatile oil of buchu.

Uses: Diaphoretic.
B. Bicyclic terpene ketones
Fenchone

Source: Fennel oil.

Uses: Fenchone is used in preparations for room sprays and bath preparations.
Camphor
62
Source: Camphor oil, which is obtained from the wood and leaves of the camphor
tree (Cinnamomum camphora).

Methods of isolation:
1. Cooling of volatile oil….Deposition of camphor occurs.
2. Camphor may also obtained from volatile oils by formation of insoluble complex with
strong acids e.g. 80% sulphuric acid.
3. Isolation from camphor trees:
The chipped wood of the camphor tree is distilled with steam and the distillate is
received in special chamber where camphor will solidify on its wall on cooling. The
solidified camphor is purified by mixing with soda lime, sand, charcoal and subjected
to sublimation, where camphor will sublime in pure state. It is pressed in cubes or
plates for exportation.
4. Most of the camphor of commerce is obtained synthetically from the α-pinene of
-
turpentine oil. Synthetic camphor is racemic, which contains CI ions due to the
method of preparation. Test for Cl - indicates its origin, natural or synthetic.

To distinguish natural camphor from synthetic:

Camphor is warmed with 1% vanillin/HCI. Natural camphor develops a yellow colour,
on further heating gives a blue, green and finally a presistant indigo blue colour is
obtained. Sytnthetic camphor gives no reaction (only a yellow-colour). Very pure
camphor gives negative colour reaction which indicates that the colour reaction is
caused by slight contamination of the camphor with constituents of camphor oil.
Natural camphor is dextrorotatory while synthetic is racemic.

Uses: Locally camphor is weakly antiseptic, mildly anesthetic, and rubefacient when
rubbed on the skin. Camphor is a pharmacetuical necessity for flexible collodion and

63
camphorated opium tincture.

Separation of mixture of borneol and Camphor:

1. The mixture is heated with phthalic or succinic anhydride. Borneol will be esterified
(acid phthalate or acid succinate), add sodium hydroxide solution and extract with
ether. The borneol acid ester goes into the alkaline solution, while camphor is
extracted by ether.

2. The benzoic and stearic esters of borneol are sparingly steam volatile, so convert
borneol into its stearic or benzoic ester and remove camphor by steam distilIation.

3. Camphor is converted to its oxime, and the oxime is dissolved in sulphuric acid. The
borneol is removed by extraction with ether.

Determination of aldehydes and ketones

1. The bisulphite method:

A measured quantity of the oil is shaken with hot aqueous solution of sodium
bisulphite, a water soluble additive compounds is formed. the non-aldehyde portion
of the separates as an oily layer. which can be measured. Using cassia flask.

2. The neutral sulphite method:

In this method neutral sulphite solution is used and sodium hydroxide is liberated as
the reaction proceeds and this must be periodically neutralized with acid to permit the

64
reaction to go to completion. This method is specifically of importance for the determinant
of carvone in spearmint, dill and caraway oils.

3. The hydroxylamine hydrochloride method:

This method based on reaction of the aldehydes and ketones with hydroxylamine
HCl giving the oxime with liberation of HCI equivalent to the aldehyde or ketone
content. This HCl could be titrated against alkali.

5-Phenols
Phenols are one of the most important constituent of volatile oils. Some oils e.g.
thyme, owe their value in the pharmaceutical field almost entirely to the antiseptic and
germicidal properties of their phenolic content. Other oils e.g. clove oil is most popular
flavouring agent.

General methods of isolation:

1 - Phenols are sufficiently acidic (with few exception) to form water soluble salts with
dilute aIkali solution (3-5%) caIled, phenolates. This property is used for the separation
of phenols from non-phenolic constituents. Thus the oil is shaken with dilute aqueous
alkali, the aqueous layer containing the phenol is separated from the oil, acidified
and the regenerated phenol is isolated either by steam distillation or extraction with
ether.

General characters of phenols:

1. Phenols react with bromine (substitution reaction) evolving HBr. The resulting
crystalline bromide is useful for identification.
2. Phenols react with ferric chloride giving a highly coloured iron compounds.
3. The phenolic group undergoes many reaction similar to those the alcohol group,

65
yielding the same type of reaction products useful for identification purpose e.g. reactions
with acetic anhydride, phenylisocyanate, benzyl chloride, p-nitrobenzoyl chloride.
Several of the common phenols may be condensed with phthalic anhydride to produce
phthalein e.g. thymol phthalein.

Thymol

Source: Thyme oil.

Colour reaction of thymol:
1. Dissolve crystal of thymol in glacial acetic acid, add drops of H2SO4 and drops of
HNO3, the solution shows a deep bluish green colour.
2. When thymol is fused with phthalic anhydride and conc. H2SO4 it develops a strong
violet red colour, and in dilute alkaline solution it develops an intense blue colour
(thymolphthalien).
3. When thymol is heated with 10% NaOH, a clear colourless or pale red solution is
formed, on addition of few drops of chloroform and shaking the mixture, a violet
colour is produced.

Uses: Thymol possesses disinfectant and antibacterial properties It is used in mouth

washes, gargles and other oral preparations. It is also used as a gastro-intestinal
disinfection. Thymol also serves as starting material for preparation of synthetic
menthol.

Carvacrol

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Source: Oil of Thymus capitatis.
Colour reaction: Carvacrol in a very concentrated a!coholic solution develops a
green colour.
Uses: Carvacrol like thymol, is powerful antiseptic and germicidal in oral
preparations.

Eugenol

Source: Oil of clove.

Uses: Eugenol is widely used in pharmaceutical preparations, as well as for
flavouaring all kind of food products. It is used in toothache remedies, because of the
local anaesthetic property.

Determination of phenol content:

Volatile oils that contain phenols when shaken with solutions of sodium or potassium
hydroxide diminish in volume because of the ready solubility of the phenol
constituents in alkali, the non phenolic portion of the oil remains undissolved.

Use is made of this property of phenols in the official assay methods. In these
determination a cassia flask is used. In this flask, an accurately measured volume of
the oil is shaken vigorously with a portion of the caustic alkali, and a further quantity
of the alkali is then added to bring the undissolved oil into the graduated neck of the
flask when the liquid have separated and become clear, the volume of the oil
insoluble in alkali is read off, the volume of the phenol being determined by
difference.

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 6-Phenolic ethers
Most of phenolic ethers possess a powerful aromatic odour and flavour (e.g.
anethole). Several phenolic ethers can be isolated by cooling the oil, or the proper
fraction to a low temperature.

Characters of phenolic ethers:

1-They are very stable neutral compounds and do not react with alkalies.
2-When they are heated with hydrobromic or hydroiodic acid, they are converted into
phenols.
3-They do not undergo the reactions typical for phenols and so their identification is
accomplished either through conversion to phenols or through the reaction
products typical of aromatic ethers.

- Phenolic ethers can be identified by formation of derivatives which result from

reactions such as: bromination, nitration, oxidation or reduction with
chlorosulphonic acid reagent, to give sulphonyl chlorides, which yields
sulphonamides when heated with ammonia or ammonium carbonate.
4- Many phenolic ethers form crystalline derivatives with picric acid, mercuric acetate,
trinitrobenzene. These crystalline derivatives serve for the identification purposes.

Anethole

Source: Anise oil.

Uses: Anethole is used widely for flavouring food and beverage products, also for
flavouring pharmaceutical preparation as mouth washing and gargles.

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 Anisole

(Phenyl methyl ether)

Source: Oil of wintergreen.

Safrole

Source: Oil of sassafras.

Uses: Safrole is widely used for flavouring of certain beverages and pharmaceutical
preparations. The principal use is the manufacture of heliotropin.

Myristicin

Source: Oil of nutmeg.

Apiole

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Source: Parsley seed oil.

Zeisels method for quantitative methoxyl determination:

This method depends on the splitting off the methyl group by constant boiling with
hydroiodic acid resulting in the formation of methyl iodide The volatile methyl iodide is
transferred quantitatively (by means of stream of a carrier gas as CO2) into receiver
containing alcoholic solution of AgNO3. The methyl iodide reacts with AgNO3,
forming the double salt AgI. AgNO3 (soluble) which is decomposed with dilute HNO3
into AgI (ppt.) and this is separated by filtration and weighed.

7- Oxides
Cineole

(Eucalyptol , cajuputol, 1:8 cineole)

Source: Eucalyptus oil (30-70%) and cajuput oil (40%).

 Cineole is separated by fractional distillation, the fraction b.p. 170-l80ºC being

childed and the crystalline cineole collected (m.p.1ºC).
 Pure cineole form number of crystalline adducts with halogen acids HCl & HBr,
O-cersol, resorcinol and phosphoric acid. Some of these adducts have been used
in the purification of cineole.

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  The oxygen atom of cineole shows basic character. The adduct with hydrogen
chloride, for example, is an oxonium chloride , this and other salts are hydrolysed
by water.
H
(R – O+ – R) Cl -

Microscopical test for cineole:

Cineole forms with resorcinol and hydroquinone a crystalline compounds of special
shape under microscope (prisms or rhomboids).

Uses: Cineole is used internally as a stimulating expectorant, locally it is employed for

its antiseptic effect in inflammations of nose and throat and in certain skin diseases.
It is some times used by inhalation in bronchitis.

Determination or cineole:
The method is based on the formation of a solid, loose molecular compound of
cineole and phosphoric acid by the lowering of the freezing-point of the mixture, from
which the cineole is regenerated by the addition of warm water. The volume of cineole
liberated is measured in cassia flask.

8-Peroxides
Ascaridole

Source: Chenopodium oil.

Separation: Isolated by fractional distillation in vacu , followed by purification by
crystallization (m.p. 2.5°C).
 Ascaridole when heated to 130-150ºC it decomposes with violent explosion.

71
  It is also decomposes with explosive violence by the action of sulphuric,
hydrochloric or nitric acid.
 It is a peroxide and liberated I2 from KI in acetic acid solution.
Uses: Ascaridole is used as anthelmintic, particularly useful against hookworm,
roundworm and dwarf tapeworm. It serves in many medical and veterinary
preparations.

Determination of ascaridole content

The method is based on the oxidising action of the peroxide radical on acidified
solution of KI and titration of the liberated iodine with sodium thiosulphate. It is very
important to maintain the low temperature (-3°C) because reaction is vigorous and
explosive and the liberated iodine is capable of being absorbed by the unsaturation in
the oil.

9- Nitrogen compounds

9- Nitrogen containing compounds

Indole

Source: Occurs in several volatile oils (small amounts) derived from flowers e.g.
jasmine, neroli.
- Indole possesses powerful, presistant, disagreable fecal odour, but if pure and
very diluted, it has a flowary, heavy odour.

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Uses: It is widely used, but in treaces, for the compounding of perfumes.

Skatole

3-methyl Indole

Uses: Used as a modifier and fixative of perfumes.

10- Sulphur containing compounds

Sulphides occur quite frequently in plants, propably in the form of glycosides which
are decomposed by the process of distillation

1. Hydrogen sulphide (H2S)

It occurs as a decomposition product of anise and caraway oils during
distillation.

2. Carbon disulphide (CS2)

It occurs as a decomposition product of mustard oil.

3. Dimethyl sulphide (CH3)2S

It occurs as a decomposition product of American peppermint oil and must be
removed by rectification in order to improve the odour of the oil.

4- Diallyl disulphide :
It occurs in oil of onion and oil of garlic.
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 5- Allylisothiocyanate
CH2=CH-CH2-N=C=S
It occurs in glycosidal combination as sinigrin in the seeds of black mustard (
Brassica nigra) .Sinigrin, on hydrolysis with enzyme myrosin, gives allyl
isothiocyanate, d-glucose and potassium hydrogen sulphate.

Method of preparation
1-The black mustard seeds are defatted by expression.
2- The marc left after expression is macerated in water at 37- 40ºc for about 5
hours to induce enzymatic action, where the glycoside is hydrolysed.
3- After hydrolysis, the mixture is subjected to steam distillation.
4- The oily layer ( heavier than water ) is collected, dried over CaCl2 and then
rectified by fractional distillation.

* Allylisothiocyanate is also prepared syntheticlly by the distillation of allyl chloride

with sodium thiocyanate in alcoholic solution.

Uses: Allylisothocyanate is used in flavouring of different kinds of food products. It

is used externally in some pharmaceutical preparations e.g. plasters and as
rubefacient and vesicant.

Determination of allyl isothiocyanate content

The official method is based upon the reaction of the isocyanate radical with silver
nitrate. The excess silver nitrate solution is determined by titration with standardized
ammonium thiocyante solution in the presence of ferric ion.
- The alcoholic solution of the oil is treated with ammonia and excess standard silver

74
 nitrate.
- The mixture is heated on water both; ammonia reacts with allylisothiocyanate
forming allyl thiourea. This reacts quantitatively with Ag2O preciptating Ag2S.
-The mixture is then filtered, acidified with nitric acid and the excess of silver nitrate
is titrated against ammonium thiocyanate using ferric sulphate as indicator.

11- Lactones
Bergaptene

(5-methoxy furanocoumarin)
Source: Bergamot oil.
Uses: Bergaptene is used for treatment of Leukoderma.

References

1- Guenther, E. ( 1952 ): The Essential oils ( vol. I-VII ), Van

Nostrand, N.Y.
2- Formacek, F.& Kubeczka, K. (1982) : Essential oils analysis, John
Wiely & sone (London) .
3- Baerheim, A.& Scheffer, J. (1985) : Essential oils and Aromatic
plants, Martinus Nijhoff (London) .
4-Trease, G. & Evans, W. (1998) : Phormacognosy, 14 ed., Baillier
Tendall (London) .
5- Official methods of analysis of the AOAC .
6- The extra Pharmacopeia 32 ed., "Martindale" (1999) .
7- De Smet, P. and Keller,. (1992) : Adverse effects of herbal drugs.

75
Springer-verlag. Berlin .
8- Pinder, A. (1960) : The chemistry of the terpenes, chapman & Hall
LRD .
9- Goodwin, T.W. (1971) : Aspects of Terpenoid " Chemistry and
Biochemistry, Academic press " .
10- Egyptian Pharmacopoeia (1984) .
11- WHO Monographs of Selected Medicinal Plants WHO, Geneva,
volume I (1999).
12- Pharmaceutical Preparations Registered in Norway (Fellesk
Katalog) (1992/1993) .
13- US National Formulary, (2000) .
14- British Parmaceutical 12th Codex (1994) .
15- Remington, 20, (2000) .
16- The Merck Index – 12th (1996) .
17- Pharmacognosy, V.. Tyler, L.R. Brady and J.E. Robbers, Lea
and Febiger, 9th ed. (1988) .
18- Medicinal Plant Constituents, S.I. Balbaa, 2nd ed. (1976) . Central
Agency for University and School Books .
19- H.D. Reuter, and A. Sendl, in "Economic and Medicinal Plant
Research, Academic Press, Vol. 6, 55 (1994) .
20- Principles of Medicinal Chemistry, W.O. Foye, T.L. Lenke, and
D.A. Williams, 4th ed., Alla and Febiger Book, Williams and Wilkins
(1995) .
21- Burger's Medicinal Chemistry and Drug Discovery Manfred E.
Wolf (ed), John Wiley and Sons, Fifth ed.(1996) .

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