11

CHAPTER I

INTRODUCTION
 12

Spices are important food ingredients traded in the world market

that are valued for their aroma and taste. They are fragrant and
pungent plant products that add zest and tang to a variety of
food preparations. Hajority of the spices originate in the
tropical regions of the world. India is the land of spices and
several spices are grown in this country. Of the 63 spices grown
in India, cardamom, ginger and turmeric had their origin in this
country. Spices such as chillies, cinnamon, clove, nutmeg, mace
and vanilla were introduced here by invaders.

1.1 HISTORY OF SPICES

Spices have been sought after since early times for the flavour
they impart to food. The early Indian scriptures dating back to
6000 BC mention the use of spices in medicine, as flavorants and
as preservatives, while the Bible refers to their use as
perfumes. Ancient Greeks and Romans used them as cosmetics,
aphrodisiacs and as presents for Gods while Chinese used spices
as medicines and as healing compounds 5000 years ago. The
1
Chinese herbal', the first medical treatise written by Shen Nung
about 2700 BC listed more than 100 spices and herbs. Spice
ownership was often associated with wealth and countries which
controlled spice trade were the most powerful. The search for
spice rich lands of the east, mainly India, gave impetus to the
discovery of new continents and trade route and the merging of
the eastern and western civilizations. South West coast of India
was one of the most important centres for spice trade from where
spices were exported to Europe and Middle East. Towards the end
of 19th century and beginning of 20th century, USA became an
important centre for spice trade. The history of spices has been
reviewed by Parryl and Rosengarten2.
 13

1-2 SPICE TRADE

1.2.1 International

Several tropical countries of the world grow spices as a crop.

India mainly produces black pepper, cardamom, ginger, turmeric
and chillies. Other major spice producing countries are Pakistan
(Chillies), Zanzibar (Cloves), Malagasy Republic (Clove and
Vanilla), Jamaica (Ginger & Pimento), Grenada (Nutmeg),
Sierralone (Ginger), Nigeria {Ginger), Sri Lanka and Seychelles
(cinnamon l, Indonesia ( Dlack &. white pepper) and Malaysia (Black
& white pepper). USA constituteH Hingle largest spice market in
the world with maximum numoer of spices being imported to that
country followed b~r Germany and Japan. In recent years, Middle
East, New Zealand, Australia and Singapore have also become
important markets for spice trade.

Black and White pepper are invariably the principal spices

imported in terms of both volume and value. Pepper alone
accounts for 30% of the international ~rade and togeth~~ with
clove, cardamom, nutmeg and mace it represents 70% of the world
Lealie iu spices3. The nine spices namely pepper, cinnamon,
clove, cardamom, nutmeg, mace, cassia, ginger and pimento account
for about 90% of the total international trade.

1.2.2 National

India is one of the largest producers and exporters of spices.

Due to the varying climatic conditions available, almost all
spices are grown in this country. India holds prime position in
the world \vi th respect. t.o pepper and cardamom. Other major
spices such as ginger, turmeric and chillies are also grown here
to a large extent. South India commands sole or near monopoly in
the production of pepper, cardamom, ginger and turmeric, while
North India contributes sizeable quantities of other spices.
Kerala state ranks first in India in the production of pepper
and cardamom contributing 96% and 70% respectively of the total
 14

amount for these spices produced in the country4, 25% of the

total ginger grown in India also comes from Kera1a. Cinnamon,
clove, nutmeg and mace are grown in Kerala but do not have
sufficient production to be exported. They are used mainly for
domestic consumption.

Spices export from India have registered an impressive growth

during the lasL five years. Of the worlds Lotal trade in spices
in 1991-92 estimated approximately at 4 lakh tonnes and valued at
45 billion, India contributed 1.3 lakh tonnes estimated at 3620
million. In terms of volume of trade, India's share in the
global market is thus about 33%4. East Asia accounted for 49% of
total quantity of spices exported and 36% of the value realized.
Fourteen countries namely USA, Bangladesh, Srilanka, USSR, UAE,
Japan, UK, Singapore, Saudi Arabia, Germany, Canada, Pakistan,
Italy and Mexico account for 85% of the total spice export
earnings of India4.

Pepper, the most important spice traded in the world market both
in quantity and value, is generally available in 3 mai~ forms
namely black, white and green pepper. Black pepper is obtained
by sun drying mature gree11 pepper berries, while fully ripe dried
fruits devoid of pericarp form the white pepper. Although black
and white pepper are the traditional forms of pepper available in
the market, green pepper has of recent years become an
important product valued mainly in the Western markets as a
garnishing spices for its appearance and delicacy. Table 1 gives
the export of green pepper in brine for the last three years5,

TABLE 1. EXPORT OF GREEN PEPPER IN BRINE5

(QUANTITY IN TONNES AND VALUE IN RS.LAKHS)

1988-89 1989-90 1990-91

QUANTITY VALUE QUANTITY VALUE QUANTITY VALUE

211 56 325 83 1240 243

 15

Its demand is thus found to be steadily increasing. Hitherto no

attempts have been made to popularize these green pepper products
as a result of which consumption is limited to a few countries
only. Green pepper is, in general, considered to be sterilized
nature-like product that retains all the flavouring principles
within it and hence assumed to be of commercial importance.

"The term spices6 applies to any dried, fragrant, aromatic or

pungent, edible vegetable or plant substances, in the whole,
broken or ground form, that contributes to the flavour and whose
primary function in food is to contribute relish or piquancy
rather than nutrition". Spices may be classified based on the
plant parts from which they are derived 6' 7 such as
fruits (capsicum, black pepper, cardamom etc.), seeds (aniseed,
caraway, celery, coriander, cumin, fennel, fenugreek, mustard,
nutmeg etc.), Aril (mace), rhizome or roots (ginger, turmeric
etc.), leaves (bay leaves, marjoram, parsley, sage, thyme), barks
(cinnamon and cassia), floral parts (saffron, cloves, caper) and
bulbs (onion, garlic, shallot).

A condiment is one containing one or more spices or spice

extractives which when added to food, after it has been served
enhances the flavour of food. Seasonings are mixture of
spices/spice extractives added to food either during its
manufacture or in its preparation before it is served.

1.4 TYPE OF SPICE PRODUCTS

Bulk of the spices are generally shipped in the whole dried form
to importing countries where they are further processed into
various products depending on the end use. Spice extractives such
 16

as the essential oils and oleoresins are also imported in larg•

quantities. Some of the other value added products available i1
the market are aquaresins 1 encapsulated ·spice oil or oleoresin:
and dry soluble spices31617,

1.4.1 Ground spices

There is considerable volume of international trade in groun<

spices. It is available in variety of different grinds in th•
market. H ix tures or blends of numerous spices in vary in!
proportions 1 known as curry powders, are also available in th•
rna r k e t and are used as s e as o n i n g s . The ad van t a g e s an<
disadvantages of handling and use of ground spices are summarize<
in literature3.

1.4.2 Essential oils

Essential oils or in other words the volatile oils may be define<

as natural odoriferous organic compounds occurring in, OJ

isolated mostly f:tom plant materials and in certain cases als<

from animal products. Essential oils are the principal but no1
the sole flavouring constituents of the spices.

Advantages of using spice essential oils are that they arE

generally uniform in flavour quality regardless of the quantit3
present. They are sterile, free from extraneous matter, solublE
in liquid fats or oils, emulsifiable in other liquid solutionsl
stable under good storage conditions and represent 98% saving ir
weight and storage space.

1.4.3 Oleoresins

Oleoresins are viscous and resinous concentrated extracts of

spices containing all its flavouring ingredients in a specifiec
solvent. Essentially, oleoresins consist of essentia]
oil,soluble resins and other related non volatile material~

inherently present in the spice. The nature of solvent used for

its preparation decides the quality of oleoresin to a great
extent. Solvents generally used are acetone, ethanol, hexane and
ethylene dichlorides and mixtures thereof.

Oleoresins have the advantage in that they have uniform,

standardized and complete flavour equal to natural spice and are
free from bacteria, molds, filth, enzymes and other antioxidants.
They are however highly concentrated and viscous creating
problems in weighing and gives spots when incorporated into food
matrix. New type of oleoresins such as super resins and aqua
resins have recently been introduced that have more specific
advantages to the end product.3

1.4.4 Other products

Other spice products that are available in the market are

encapsulated oils and oleoresins dry and liquid soluble spices,
spice essences and emulsions, spice decoctions, spice pastes and
concentrates and spice salts. These products have been developed
according to the end uses.

1.5 PROPERTIES AND USES

The use of spices as flavouring agents, as antioxidants and as

preservatives were known to the ancient people since early
times3. Their antimicrobial and medicinal properties were made
use of in ancient systems of medicine. The ayurv~dic system of
medicine prevalent in ancient India mentions the wide use of
spices in curing several diseases8. Even today, spices are
highly.valued and widely used for these properties. In addition
to the above properties, spices find application in perfumery and
cosmetics. Some of the important spice oils namely that of
cardamom, cumin, celery and nutmeg are being used in different
types of perfumes. The oil of cinnamon, fennel, nutmeg etc. are
used for scenting soaps, dental preparations, hair lotions etc.
 18

The above properties are attributable mainly to the essential oil

components and also certain other non volatile constituents suet
as phenolics which have been highlighted in section 1.6.

1.6 CHEMICAL CONSTITUENTS OF

SPICES

Chemic a 1 constituents of spices in general. are starch,

carbohydrates, proteins, fats, vitamins, alkaloids, essentiaJ
oils, and phenolic compounds besides pigments and other minoi
constituents. Table 2 lists the general chemical composition o1
the six commonly used spices6, Spices are used in low amounts ir
foods and hence these constituents do not have any significant
role in increasing the nutritive value of foods to which they are
added. The essen t. ial oils, on the other hand, impart aroma, anc
the non volatile constituents especially polyphenols, anc
related compounds such as alkaloids that contribute to the taste
and pungency together play a major role in contributing ·to the
flavour of spices for which they are highly valued in the worl~

market.

1.7 CONCEPT OF FLAVOUR

Flavour may be defined as a combination of odour, taste,

texture,temperature and psychological factors (appearance, colour
etc. )3, Basic tastes like sweet, bitter, sour and salty and
specific sensations related to taste like cooling, burning,
pungent, astringent or biting e f fee ts are encountered . in
differeut spices. The aroma properties of a substance exert the
greatest influence on what is understood as flavour. Essential
oils present in spices are responsible for their characteristic
aroma. Several chemical compounds either individually or in
combination contribute to the typical aroma of the· spice
TABLE 2. APPROXIMATE COMPOSITION OF PEPPER, CINNAMON, CLOVE, CARDAMOM, NUTM! t\ND MACE6

(per 100 g edible portion)

WATER FOOD ENERGY PROTEIN FAT TOTAL FIBER ASH Ca Fe Mg P K Zn Niacin VIT ASCORBIC
(g) (Kcal) (g) (g) CARBOHYDRATES (g) (g) A ACID
(g) (mg) ( IU) (mg)

BLACK PEPPER 10.5 255 11.0 3.3 64.8 13.1 4.3 437 29 194 173 1259 1 1 190

WHITE PEPPER 11.4 296 10.4 2.1 68.6 4.3 1.6 265 14 90 176 73 1

CINNAMON 9.5 355 3.9 3.2 79.9 24.4 3.6 1228 38 56 61 500 2 1 260 23

CLOVE 6.9 323 6.0 20.1 61.2 9.6 5.9 646 9 264 105 1102 !3 1 1 530 81

CARDAMOM 8.3 311 10.8 6.7 68.5 11.3 5.8 383 14 229 178 1119 7 1

NUTMEG 6.2 525 5.8 35.3 49.3 4.0 2.3 184 3 183 213 350 2 1 102

MACE 8.2 475 6.7 32.4 50.5 4.8 2.2 252 14 163 110 463 2 1 800
 20
essential oil.

Two terms namely "aroma character impact" compound ( s) and

"contributory aroma" compound ( s) are frequently used in recent
years to define the "top aroma" note of a given spice9. The term
"aroma character impact" compound(s) refers to those compound(s)
that actually contribute to the typical aroma of a particular
spice and whose absence results in the non-acceptability of the
product by the consumer. "Contributory aroma" compound ( s), on
the other hand, are those compound(s) that stimulate the typical
aroma to a more pleasant and acceptable note. They thus help in
determining the quality of a spice at hand. These two classes of
compounds together give rise to the "top note" of the given spice
(Table 3) 10. Study of the essential oil composition thus helps
in understanding the chemical compounds responsible for
characteristic "top aroma" of a particular spice.

Several classes of compounds contribute to the taste sensation3.

For example piperine in black pepper, eugenol in clove,
isoeugenol in nutmeg and capsicin and dihydrocapsicin in red
peppers all contribute to the pungency, biting or burning effects
of these spices. Phenolic compounds, such as phenolic acids and
flavonoids in glycosidic form on the other hand, are known to
contribute to the astringencyll and bitterness12. Polymeric
phenols such as tannins (as in cinnamon) are found to be
astringent while monomeric to tetrameric phenols are mostly
bitter13.

When spices are intended for direct use in the whole or ground
form, the appearance is of primary importance to the buyer14.
Many individual factors contribute to the total perception of the
appearance of a food product. Colour and texture are two such
important factors and they play a major role in evaluating the
acceptability of foods by the consumer. While natural occurring
pigments like anthocyanins, curcumene and carotenoids contribute
to the colour of various spices, several phenolic compounds
present in certain spices undergo enzymic or non-enzymic
~---------- ---

TABLE 3. AROMA CHARACTERISTIC, AROMA CHARACTER IMPACT COMI NDS

AND CONTRIBUTORY AROMA COMPOUNDS OF SOME COMMONL) 5ED
SPICEslO

Spice Aroma Aroma Character Contrib ry

Characteristics impact compounds aroma c1 )Unds

Coriander Lemon peel, Linalool, a-pinene

leaves aromatic, spicy 13-pinene

Fennel Anise-like Ani thole, fenchine Fenchon

Ginger Aromatic, a-Zingiberene, Gingero

Pungent, spicy, 13-Zingiberene, borneol
camphoraceous geraniol,
zingirone

Mango ginger Raw mango-like cis-Ocimene, a-pinen

Car-3-ene

Mustard Slightly acrid Allylisothiocyanate

Turmeric Pungent, spicy Turmerone, borneol

 22

oxidation to produce coloured polymeric products. Formation of

the latter pigments is controlled by the level and nature of
phenolic compounds, the presence of oxygen and the amount of
enzyme activity. Thus the natural pigments and polyphenols of
spices are important constituents that decide their overall
flavour quality.

The task of identifying flavour compounds in foods is complicated

by the fact that they are present in low concentrations, show
wide variation in their qualitative and quantitative distribution
and undergo changes during isolation, storage and analysis. The
lower sensitivity of available instrumental methods compared to
human olfactory system, together with the difficulty in assessing
the importance of each chemical compound to the overall flavour
profile also limits their study.

1.7.1 General methods of isolation and concentration of flavour

compounds

The first step in the isolation of flavour compounds from· foods

involves crushing, homogenizing, blending or extracting them in
some manner with minimum loss in flavour components. Fresh foods
contain active enzymes in their tissues that can alter the
flavour profile once cellular disruption has occurredl5, Hence,
inactivation of enzymes is essential. Homogenizing with methanol
with or without thermal process are often employed for this
purpose16,

Those methods which are frequently used in recent years for the
isolation and concentration of flavour compounds from spices are
highlighted below.

1.7.1.1 Isolation and concentration of volatile flavour compounds

1.1.1.1.1 Steam distillation

0f t he s eve r a l me t hods c u r rent l y in pract i ce f o ·r t he

 23

isolation/concentration of volatile aroma constituents of spices

(Table 4), steam distillation in its various forms is the most
frequently used technique 17. However, this method suffers from
the draw back that there may be losses in highly volatile aroma
compounds that may undergo chemical changes due to
thermal treatment. Steam distillation under reduced pressure, on
the other hand, greatly reduces many of the disadvantages and
gives a higher yield of oil. The aqueous condensate containing
the volatile flavour compounds obtained above is usually
extracted with appropriate solvents and the solvent removed by
distillation under reduced pressure to yield the aroma
concentrate.

1.7.1.1.2 Simultaneous distillation-extraction

An isolation procedure that provides for efficient recovery of

volatile flavour compounds is the use of a simultaneous
distillation-extraction apparatus originally developed by
Nickerson and Likens18, The extracting solvent generally used in
this method is either diethyl ether or pentane. In this method
the volatile flavour compounds along with steam simultaneously
come in contact with the solvent vapour and subsequently separate
into aqueous and sol vent phases in the condenser. The aqueous
and the solvent phase return to their respective flasks thereby
maintaining a continuous operation.

It has the advantage of greater efficiency and lower extraction

time compared with conventional steam distillation. Loss of
volatiles during concentration is minimized due to lower amounts
of solvent used. Short contact time with aqueous phase also
reduces the risk of changes in flavour compounds during
extraction. The extract thus obtained is usually concentrated
either by distillation in a vigreux column or by removing the
solvent with a slow stream of nitrogen to obtain the aroma
concentrate. Introduction of steam in the distill.ing flask or
the operation of distillation-extraction unit under reduced
pressure are some of the recent modifications of· this
 24

TABLE 4. METHODS FOI?. ISOLATION AND CONCENTRATION OF VOLATILE FOOl

CONSTITUENTs17

Distillation

Flash distillation
Atmospheric pressure
Reduced pressure

Steam distillation
Atmospheric pressure
Reduced pressure

C02 distillation

Vacuum fractional distillation

High Vacuum distillation

Degassing
Molecular distillation

Other Types

Gas entrainment
Open system
Closed system

Distillation of aqueous and nonaqueous extracts

Lyophylization (freeze drying)

Retention of volatiles

Freeze concentration
Zone melting

Adsorption
Charcoal
Silica gel chromatography
Dry column chromatography

Extraction

Derivative formation
technique19,20,

Thermally induced chemical changes, however, can occur in

simultaneous distillation-extraction method of isolation so that
care must be taken for artifact formation during this process.

This technique alLhough widely used for spice aroma research in

several laboratories has not so far been commercialized due to
l ts sophist lea t ion in fabr ica Lion and operation. However, this
technique seems to be a novel method for almost total recovery of
essential oil from a variety of spices.

1.7.1.1.3 High vacuum distillation

High vacuum distillation or in other words closed system high

vacuum transfer method is a milder technique than the above two
distillation methods and is gaining importance in the isolation
and concentration of volatile flavour compounds from natural
products. The potential loss of very low boiling volatiles is
minimized and the formation of artifacts as observed i~ many
other distiliation procedures is eliminated. Various types of
vacuum distillation assemblies have been developed21,
Bandyopadhyay et al22 reported a simple closed system high vacuum
distillation apparatus for the isolation and concentration of
true aromatic principles of various natural products. The
assembly is a closed system high vacuum transfer of volatiles
from the samples and operated at or below ambient temperature.

Recovery of low boiling volatiles by this method is particularly

good. The high vacuum requirements however, limit this method to
laboratory scale isolations. A commercial method known as
WURVAc17 process for the stripping of volatile flavour compounds
from fruit juices using short thermal treatment under vacuum is
practiced in fruit juice industry. This technique has so far not
been applied in the spice industry to obtain spice oils.
 26

1.7.1.1.4 Solvent extraction

(a) Conventional

Most flavour compounds exhibit substantial solubility in organic

solvents such as diethyl ether, dichloromethane, pentane etc.
Oleoresins are often extracted from spices using polar solvents.
Non-fat containing foods may be solvent extracted to obtain a
flavour isolate. Batch extraction23 and liquid-liquid
extraction24 are the two methods generally used. However, by
these procedures various non-volatile constituents are also
extracted from plant products along with essential oils, which
often pose problems for the subsequent recovery of the desired
essential oils. Extraction of essential oils and oleoresins at
low temperatures using Freon became a commercial proposition for
handling bulk materials because of its non-inflammable, non-
explosive and practically odourless properties. In recent years,
however, recognition of its toxic properties has restricted its
use in such applications.

When a fatty food containing aroma compounds is to be solvent

extracted, the volatile flavourings must be isolated from the
extract via steam distillation, molecular distillation or
dialysis25,

(b) Super critical fluid extraction

Solvent extraction of spices using super critical fluids such as

liquid carbon dioxide (super critical COz extraction) has become
an important method useful both in industrial and research
applications26,27, C02 can be liquified at high pressure by
maintaining a temperature below 3l"C and at super .critical state
(72.8 atmp at +31'C) by maintaining at one above it. The high
solvation power of COz together with its low viscosity and high
solute diffusivity at high pressures and at super critical as
well as sub critical temperatures can be conveniently made use of
for the extraction of volatile flavour compounds. Its solvation
 27

power and selectivity for holding solutes can be altered by small

changes in temperature and pressure just above critical
temperature thus providing flexibility of operation. Low
proportion of modifiers such as methanol, ethanol and acetic acid
in C02 are used for selective extraction of polar compounds28.
Liquid C02 is inexpensive, non-explosive, relatively inert and
has low toxicity. Risk of degradation of labile compounds is
also reduced ~ue to the low temperatures used so that nature-
identical products can be obtained. In addition, the recovery of
the extracted compounds is simplified because the solvent can be
removed by decompression. This technique appears to have great
potentiality in the field of spice essential oils for the
production of value-added products.

1.7.1.2 Isolation and concentration of non volatile flavour

compounds

Non volatile flavour compounds of spices particularly pigments,

alkaloids and phenolic constituents are isolated by solvent
extraction. Solvents used for extraction are chosen according to
the polarity of the compound being studied. Non polar and medium
polar solvents such as hexane, benzene, chloroform, diethyl ether
or ethyl acetate are used for extraction of aglycones,
carotenoids and compounds of medium polarity while polar solvents
(acetone, alcohol, water, or a combination of these) are used for
the isolation of glycosides and highly polar compounds.
Sequential solvent extraction with a number of solvents of
varying polarity helps in separating glycosides from aglycones
and polar compounds from non-polar ones.

Concentration of the extracts thus obtained is generally carried

out by flash evaporatio11 under reduced pressure at ambient
temperature or at 40"C,

The glycosidically bound aroma compounds constituted of

glycosides of aliphatic, monoterpene and sesquiterpene alcohols,
aldehydes and acids are also known to be present in several
 ·28

natural products including spices. A varying amount of bound

aroma compounds have also been reported in g inger2 9, grapes30,
Prunus spices31 and blue berries32, These bound volatile flavour
compounds cannot be isolated by the normal isolation methods
unless they are released either enzymatically (~-glucosidase) or
chemically (acid hydrolysis) from spices. Higher yields of aroma
concentrate representing true aroma of the spice can thus be
obtained.

1.7.2 General methods of separation

The aroma and flavour compounds of spices are a complex mixture

of several organic compounds varying over a wide range in their
physical and chemical properties. The task of identifying these
constituents either individually or as a class is thus a
difficult proposition unless they are subjected to separation.
Several methods of separation are known, of these the ones most
commonly used are described below.

1.7.2.1 Separation of volatile flavour compounds

The essential oil constituents of spices vary widely in their

volatility, boiling point and functional groups and could be
classified as terpenoids, phenols, acids, lactones, carbonyls,
esters, ethers etc. Terpenoids mainly monoterpenoids and
sesqui terpenoids are the major constituents of spice essential
oils. They are mainly acyclic, monocyclic, bicyclic and
tricycli'c terpenoids. The wide range of functional moieties
found in mixtures of aroma volatiles influence both the
polarities and the acidic/basic properties of individual
molecules. These compounds need to be separated at least into
classes or individually before their identification.

(a) Preliminary fractionation

Separation of flavour volatiles into broad chemical classes by

fractionation on a silica gel column33 using solvents of varying
 29

polarity facilitates preliminary fractionation. _Not only many

fractions free from major interfering compounds can be obtained,
but also a considerable concentration of minor components of
sensory importance may be acl1ieved in the individual fractions.

Separation into acidic, basic and neutral fractions also serve to

reduce the complexity of the concentrates34, Fractional
distillation of spice aroma concentrates under reduced pressure
using different types of fractionating columns are commercially
used for the recoveries of aroma fractions having perfumery
values. Chemical methods of fractionation such as derivative
formations, although less frequently used, are also helpful.

(b) Gas chromatography (GC}

Technique of GC is of primary importance in flavour analysis35,

It is an indispensable tool for the separation, fractionation and
purification of complex mixture of volatile flavour compounds.
The main factors for achieving a successful separation are the
type of column, the nature of stationary phase, carrier gas flow
rate, temperature, sample load and type of detectors used.
Chromatographic columns are usually of two types36. eg. Packed
column and open tubular or capillary columns. Higher resolving
power, shorter analysis time, lower detection limits and
reproducible results are some of the advantages of capillary
columns over packed columns. Capillary columns, however, have
limited sample loading capacity compared to packed columns.
Selection of a column depends on the nature of compounds to be
separated. It is generally found that the use of nonpolar or
slightly polar (e.g. silicon oils) stationary phases along with
programming37 of column temperature ranging from ambient or
subambient t~mperature to as high as 250"C serves to provide the
most satisfactory separation. Technique of programmed cryogenic
temperature38 GC is also used to enhance sample separation.

Two dimentional ac39,40 involving passing a section of one GC run

over a second column of different polarity, sequentially
 30

connecting two GC columns of different polarity and operating

columns at different temperatures as in SECAT ac41 and recycling
the first run on the same column as in multiple Gc41 have all
been used to improve GC-resolving power. Separation and detection
of various stereoisomers and chiral compounds is also made
possible by the use of high resolution GC (HRGC)41 equipped with
split/splitless device and having a fused silica capillary column
or chiral column.

Of the GC detectors used in flavour research FID (flame

ionization detector) and TCD (thermal conductivity detector) are
being frequently used. FID has higher sensitivity compared to
TCD; however TCD aids in fraction collection and odour
evaluation. A recent improvement in GC involves the use of a
splitting device that enables simultaneous detection (FID) and
odour evaluation of the separated component.

Recently, super critical flu{d chromatography (SFC)42 using super

critial fluids as carrier gas has been applied in the separation
of thermally labile volatile c.;onstituents on l>ol,h packed and
capillary columns. Use of coupled (on-line) super critical fluid
extraction with capillary GC (SFC-GC) technique facilitates43
sample extraction, concentration and GC separation within one
hour and thus has great potentiality in screening flavour quality
of spice varieties.

Other chromatographic techniques for separation of flavour

volatiles such as column, paper and TLC are used in diminishing
order in recent years as they lack the efficiency, sensitivity
and resolving power compared to GC technique.

(c) High performance liquid chromatography (HPLC)

The application of HPLC technique in the separation of spice

essential oil components has been limited because of its
l imitation in the resolving power of mul t icomponent mixture and
of poor response to presently available detectors.· Low
 3i

temperature HPLC separation of monoterpene and sesquiterpene

hydrocarbons44 as well as oxygenated terpenes have been recently
reported. Thus the presently known HPLC method seems to be
advantageous in class separation of volatile terpenes which
facilitates the subsequent detailed analysis by capillary GC.

1.7.2.2 Separation of nonvolatile flavour compounds

Non volatile flavour compounds isolated from spices as mentioned

earlier (Section 1.7.1.2) have widely varying chemical structures
and polarities. Separation of these natural products from each
other is of interest since early times.

(a) Separation with selective solvents

Based on preferential solubility of certain classes of above

mentioned non vola t i 1 e compounds in specific solvents, a
pre! iminary separation can be achieved. Use of C02 extraction
technique for this purpose appears to be a promising method in
recent years28,

The separation by solvent extraction method enable a class

separation and concentration of the compound of interest from the
bulk spice samples. However, it fails to separate individual
constituents of the separated classes. More specific and better
separation techniques have, therefore, been developed.

(b) Chromatographic techniques

Adsorption techniques such.as column, paper and thin-layer

chromatography are routinely used for the isolation of
nonvolatile flavour compounds. Adsorbents such as silica gel,
cellulose, alumina, sephadex LH-20, ion-exchange resins (Dowex
and Amberlite XAD-2) and polyamide are widely used for this
purpose. Polyamides have high capacity for binding phenolic
compounds which can then be isolated by extraction with a
suitable solvent.
 32

Non volatile flavour compounds as such cannot be handled by GC

without prior modification. Volatilization of these constituents
by deri vatization is a common practice generally employed. Non
volatile constituents such as phenolic acids and their glycosides
are usually converted to their trimethylsilyl derivatives. Using
this method separation and analysis of such compounds on GLC have
been carried out in several spices and plant foods45, 46. HPLC,
on the other hand, requires usually no such pretreatment and
hence is the method of choice in the analysis of such compounds.

The usual chromatographic principles employed in ihe separation

on HPLC column are partition, adsorption, ion-exchange and
exclusion chromatography. A wide variety of polar and nonpolar
compounds can be separated either on a normal phase (Stationary
phase more polar than mobile phase) or on a reverse-phase
(stationary phase less polar than mobile phase) column. Klick
and Hermann46 reported a successful separation of phenolic acids
and their glycosides by HPLC. Using HPLC technique Biacs et al47
separated 16 carotenoid pigment components of paprika (capsicum
annuum.>. A system of HPLC and supercritical rluid
chromatography ( SFC) was elaborated and developed for the
separation, identification and determination of paprika
oleoresins and associated carotenoids48, Recently, HPLC of foods
on silica bonded phases have been reviewed by Khurana49, where
the application of various bonded materials such as amino, cyano,
dial, C-18, C-8, chiral etc. phases to analyze and determine food
components such as food colour, pigments, carbohydrates, flavours
including phenolic acids etc. have been discussed.

1.7.3 General methods of identification

Physical, chemical and instrumental techniques have been widely

used in the objective evaluation of volatile oils and other
chemical constituents of spices. Although a large number of
methods are available for identification of unknown compounds
only a few are useful in flavour research. Of the physical
methods, specific gravity, refractive index, optical rotation,
 33

optical density and solubility have been used in commercial

practice. Certain characteristic features such as colour as in
turmeric, pungency as in black pepper and capsicum and
astringency as in cinnamon are routinely evaluated using
physicochemical methods. Measurement of colour intensity of the
product by colorimetry ej ther directly or by using chromogenic
reagents are the methods commonly used in such cases50,51,52,
However, these methods give a qualitative assessment of the
product. With the advent of instrumental methods such as
chromatography and spectrometry, objective evaluation of spice
and spice products has been perfected to a great extent with
respect to their quantitative composition and characterization.

The chromatographic methods as mentioned earlier (·Section 1. 7. 2)

are primarily separation techniques that are being used for the
detection and tentative identification of aroma and flavour
compounds. Internal and external standards are frequently used
in chromatographic analysis for the quantitative assessment of
aroma and flavour composition in spices. The unique feature in
both GC and HPLC is the ability to handle low amounts of ~amples

for analysis exhibiting a clear profile of the constituents and

their· l]llaljtatlvc as well as quantitative composition. However,
they have limitation in the characterization of the separated
compounds, as the identification is primarily based on retention
data. Therefore, GC and HPLC methods should be supplemented with
other instrumental methods of analysis for the structural
characterization of aroma and flavour components.

Increasing attention has been paid to on-line GC coupled to mass

spectrometer (GC-MS)53,54 and on-line HPLC coupled to MS (LC-
MS)55,56 for the identification and simultaneous characterization
of volatile and nonvolatile flavour compounds in foods including
spices. In this system, the components eluting out of the column
are directly introduced through an interface into the MS where
they are subsequently identified based on their fragmentation
pattern. MS is a method of choice for the identification of
flavour constituents in foods due to its high sensi ti vi ty ( 10-
 34

12g) and specificity. In off-line analysis the constituents

separated by GC or LC.are collected and leisurely analyzed by MS.
Computerization of the on-line assemblies including data
processing, print out and video display of the analytical results
have been well developed which reduces the time of analysis
significantly.

In recent years, supercritical fluid extraction coupled to a GC-

MS system ( SFE-GC-MS) 57 for the extraction and simultaneous
analysis of volatile flavour compounds of various materials has
been developed and this complex system appears to be a very
useful method for the quality evaluation of spices and spice
products.

Other ancillary identification methods such as infrared (IR,FT-

IR) spectroscopy, nuclear magnetic resonance (lH, 13c, 2D, COSY,
NOE, NOESY) spectroscopy and MS have been employed for the
identification and absolute configuration of the flavour
constituents separated by GC and LC.

1.7.4 Sensory analysis

Sensory analysis58 of foods relies upon human senses (odour,

taste, tactile, temperature, pain etc.) for the evaluation of
food flavours. The great sensitivity of human olfactory system
especially the sense of smell helps in detecting minute
difference in flavour that would be impossible using chemical
methods and physical instruments. Sensory analysis is used in
assessing the nature and quality of raw materials and finished
products, competitive products and development of new products.
Such sensory procedures may be classified into difference,
preference and descriptive techniques. Several studies on the
sensory evaluation of spices are reported in literature3,
Correlation of GC data with sensory techniques for the quality
evaluation of spices have been reported by various workers59,60,
This objective~subjective evaluation has been used to categorize
spice according to their quality.
 35

The general aspects of spices and spice products and the method
of recovery of flavour compounds from them has been briefly
discussed in the foregoing section. The subsequent sections
individually review the relevant literature on the commercially
important spices under the present study.

1.8 PEPPER ( .P.I.P.E.R .N".IG.RUM" L. )

Piper nigrum is a perennial woody climbing vine or shrub, 5-10 m

or more in height. Fruits called drupes or berries are dark
green and ovoid turning to bright orange and red when ripe. More
than 70 cultivars of pepper are grown in India. The berries are
harvested 6-8 months from flowering. Black and white pepper are
obtained in yields of 33% and 25% of the weight of green berries
respectively.

Black, white and green pepper are the three main products
available in the market. Processed products as mentioned earlier
are also available. Pepper is graded in the market based
primarily on three quality attributes namely appearance, pungency
and aroma/flavour quality14. These quality attributes are
primarily dependent on the intrinsic characteristics of the
cultivar grown, area of cultivation, processing methods,
condition and duration of storage and upon the maturity of the
berries at the time of harvest61,62,63.

1.8.1 Products and Uses

For the commercial production of black and white pepper64, sun

drying is the traditional method used. However, artificial
drying using smoke or modern driers are also employed. Black
pepper is used in a wide range of food stuffs particularly in
meat products both for domestic culinary purposes and in
flavoring processed food. White pepper is used in flavouring
foods when dark particles are undesirable such as in ·light
 36

coloured sauces, mayonnaise and cream soups.

Black pepper is made into a variety of grinds ranging from

1
cracked' to fine ground, while white pepper is normally made
into fine grinds64. They are extensively used for culinary
purposes and for seasoning processed foods.

Pepper essential oils are obtained from dried pepper berries and
also to a large extent from pin heads and wastes by conventional
distillation meth~ds64. Pepper oil is widely used in perfumery
and in flavourings.

The oleoresin of pepper consists of essential oil, piperine, the

pungent principle and varying amounts of non flavour constituents
like fixed oils, colouring matter, resins and sugars etc. Yield
of oleoresin varies from 6-10% in both black and white pepper; it
contains 20-25% essential oil and 35-40% piperine14,64. They are
used extensively in processed foods.

Green pepper berries in a solution of 16-20% brine and 0. 2%

citric acid with 100 ppm S02 gives a product that is free of
discolouration, gelation and berry rupture61. Dehydrated green
pepper is an another type of product that has good market
potential 55. It has high flavouring properties ( 3-5% volatile
oil and 7-9% piperine) besides the natural appeal that it imparts
to the food. It rehydrates in warm water to almost the natural
appearance within 15 min.

1.8.2 Medicinal uses

The medicinal use of black pepper in Ayurveda has been known

since early days. It is being employed even today as an
important ingredient in various Ayurvedic preparations8' 66. It
is used especially in intermittent fevers, indigestion,
dyspepsia, diarrhoea, flatulence and worm infestation. Asthma,
cough, heart troubles and pains are relieved by it. It is used
in the treatment of diabetes, piles, epilepsy, elephantiasis and
 37

filaria and is an antipoison.

1.8.3 Chemistry

Pepper berry contains essential oil, fat, phenolic.s, alkaloids,

resins, proteins, cellulose, pentosans, starch, minerals etc.
Chemical composition of world trade varieties have been
exhaustively reviewed by Govindarajan 64 who reported a wide
variation in the composition of individual chemical constituents
among different pepper varieties grown in various regions of
India. Varieties grown in Kerala had intermediate values of
volatile and nonvolatile constituents compared to that grown in
Assam and Karnataka64,

Outer skin of pepper which comprises 25% dry weight of black

pepper contains mainly fibers and some essential oil cells which
are lost in the preparation of white pepper. White pepper thus
differs from black pepper in their fibre content (3-5% compared
to 10-15% in black) and starch content (55-60% as against 35-48%
in black)14, Green pepper contained the highest le~el of
volatile oil which is proportionately related to the lower level
of starch present in green pepper compared to black and white
pepper67, The fat content of pepper was found to vary from 1.9
to 9.0% with palmitic, oleic and linoleic acids being the
predominant fatty acicts64,

Besides the major flavouring principles such as essential oil and

piperine, phenolic constituents present in pepper play a
significant role by imparting astringency and bitterness as
mentioned earlier. However, little attention has so far been
paid to the study on the nature of phenolic compounds in pepper
berries.

The chemistry of certain important constituents of pepper namely

essential oil, pungent principles and phenolic constituents are
reviewed in this section.
 38

1.8.3.1 Essential oil

Essential oil of pepper is a pale green mobile liquid becoming

viscous on ageing and gives a warm-spicy aroma. The yield of oil
can vary depending upon the type of material used, varietal
variation, maturity, processing methods and upon the conditions
and method of extraction and storage64. Lower yields are noted
in oils distilled from old and damaged pepper corns and from
pepper wastes such as husks and pin heads. A wide variation in
the yield of oil among cultivars could be noted ranging from 0.6%
to 4.1%64, Changes in volatile oil content with maturity have
been shown earlier in black, green and white peppers grown in
Srilanka67. The yield of oil was also found to depend upon the
processing methods used prior to isolation . . For example,
cryornilled pepper gave a higher yield of oil (1.84%) compared to
that ground at room temperature (1.4%)64, This could be
explained on the basis of greater release of flavour compounds
from fine grinds produced during cryomilling and a reduced loss
of low boiling compounds. Conventional steam distillation
generally used for obtaining pepper essential oils give a lower
yield of oil due to losses in volatile compounds during heat
treatment. Supercritical fluid extraction using C02 as mentioned
in section 1.7.1.1.4.b have recently been introduced for the
extraction of volatile oils from pepper68,

Composition of volatile oil was examined by several workers. As

early as 1835 Dumas69 examined the composition of black pepper
oil by classical isolation and derivatization techniques and
found a high proportion of monoterpene hydrocarbons. Later
Schreiner and Kremers70 identified sesquiterpenes and oxygenated
compounds as components of the oil. Inspite of its great
importance a detailed composition of pepper oil was not available
until the advent of GC and GC/MS techniques. Detailed analysis
of pepper oil was first attempted by Hasselstrorn et al71 who
showed the presence of terpene hydrocarbons such as a- and P-
pinenes, a-phella.ndrene, dl-1 imonene, 13-caryophyllene and
oxyg~nated terpenes namely epoxy dihydrocaryophyllene, piperonal,
 3"9

cryptone and dihydrocarveol. Ikeda et al72 further identified a-

thujene, camphene, sabinene, 6-3-carene, a-terpinene, myrcene, ~­

phellandrene, Y-terpinene, p-cymene, terpinolene and ocimene in

addition t..o t..he above compounds. Nigam and Handa73 as· well as
Lewis et al74 confirmed the presence of many of these monoterpene
hydrocarbons in Nalabar pepper oil and additionally identified
the sesquiterpenes namely a-bergamotene, a-humelene and a-
selinene. Jennings and co-workers75,76 identified several
additional compounds in Srilankan blacl\ pepper. Sesquiterpene
hydrocarbons namely 5-elemene, a-cubebene, a-copaene, 0-elemene,
a-cis-bergamotene, a-santalene, a-trans-bergamotene, [3-
caryophyllene, 0-bisabolene, 5-cadinene, calamenene,
isocaryophyllene, 1-murolene, 0-farnesene and ar-curcumene,
oxygenated compounds namely linalool, 1-terpinene-4-ol, u-
terpineol, cryptone, carvone, p-cymene-8-ol, trans-carveol,
safrole, methyl eugenol, nerolidol, myristicine, cis-and trans-
sabinene hydrate, elemol, cis-c~rveol, eugenol,
cis-p-2-menthen-7-ol, cis-p-2, 8-menthadien-1-ol, trans-
pinocarveol and a number of partially characterized sesquiterpene
ketones, aldehydes and alcohols were identified in the oil. New
constituents of oxygenated fractions of Lompong black pepper
essential oil have been reported by Debrauwere and
Verzele77. They identified forty new oxygenated compounds of
which the major compounds include butyric and benzoic acid,
butyrates, piperonal, carvetonacetone, cadinene-4-ol, linalool,
caryophyllene oxide, p-cymene-8-ol-methyl ether, a~pinene oxide,
limonene oxide, dihydro carvone, methyl carvacrol, nerol, 2-
undecanone, p-cymene-8-ol-methyl ether, 0-pinone, 1,1,4-trimethyl
cyclohepta-2,4,-dien-6-one, 1-terpinen-5-ol, 3,8(9)-p-menthadiene
-1-ol, N-formyl piperidine, 1 ( 7), 2-p-methadien-4-ol, 1 ( 7), 2-p-
menthadien-6-ol, 5,10(15)-cadinen-4-ol and [3-caryophyllene
alcohol.

It has now been shown that the pepper oil has an extremely
complex composition, composed of a variety of monoterpene and
sesqui terpene hydrocarbons and various oxygenated compounds.
Oxygenated compounds, although minor in composition, may play an
 40

important role in determining the ogranoleptic properties of the

spice14, Constituents identified so far in steam distilled black
pepper oils have been summarized in table 5. Thus a total of 102
compounds have been identified in pepper oil. The phenyl ethers
and micellaneous compounds listed in table 5 are present in low
amounts.

Qualitative and quantitative variation78 in the composition of

pepper oil obtained from different Indian cultivars is reproduced
in table 6. These studies have shown that the monoterpene
hydrocarbon and sesquiterpene hydrocarbon groups varied from
49.2% to 77.5% and 18.5% to 42% respectively, while the
oxygenated compounds varied from 2. 0% to 15.7% in different
varieties. a-phellandrene and 6-3-carene among the monoterpenes
showed considerable variations ranging from 0. 46 to 27% and
traces to 15. 5% respectively. Less pronounced compositional
changes were observed in sesquiterpene hydrocarbons and
oxygenated compounds. {3-caryophyllene was the major
sesquiterpene hydrocarbon, being least in ·Kuthiravally (9.39%)
and highest in Narayakodi (30.94%). Linalool and Caryophyllene
oxide were the dominant oxygenated compounds. Variation in the
essential oil composition among different Indian pepper cultivars
have recently been studied by other workers79,80,

White pepper has a lower oil content (0.5-1.5%) compared to black

pep p e r , however , they do not show any qua 1 it at i v e and
quantitative differences in their oil composition compared to
black p.epper81,

Very few reports are available on the volatile oils of green

pepper. Janz et a162 have reported the composition of volatile
oil obtained from fresh green and dried peppers of Sri Lankan
origin and found it to be similar to that reported for Indian
black peppers 64. They showed that except for 1-terpinene-4-ol
and {3-caryophyllene no significant variation in the composition
of other constituents occurred during maturation. Monoterpene
content always accounts for 65-70% of the oil during the ~ntire
 41
TABLE 5. CONSTITUENTS IDENTIFIED IN BLACK PEPPER OILS14

Monoterpene hydrocarbons cis-carveol

trans-carveol.
camphene carvone
'& -3-carene carvetonacetone
p-cymene 1,8-cineole
limonene cryptone
mycrene p-cymene-8-ol
cis-ocimene p-cymene-8-methylether
a.-phellandrene dihydrocarveoi
13-phellandrene dihydrocarvone
a.-pinene linalol
sabinene cis-2,8-menthadien-2-ol
a.-terpinene 3,8(9)-p-menthadien-1-ol
Y-terpinene 1(7),2-p-menthadien-6-ol
terpinolene 1(7),2-p-menthadien-4-ol
a.-thujene 1,8(9)-p-menthadien-5-ol
1,8(9)-p-menthadien-4-ol
Sesquiterpene hydrocarbons cis-p-2-menthen-1-ol
myrtenal
a.-cis-bergamotene myrtenol
a-trans-bergamotene methyl carvacrol
13-bisabolene trans-pinocarveol
o-cadinene 13-pinone
1-cadinene cis-sabinene hydrate
calamenene trans-sabinene hydrate
13-caryophyllene 1-terpinen-4-ol
a-copaene 1-terpinen-5-ol
a.-cube bene a-terpineol
ar-curcumene 1,1,4-trimethylcyclohepta-
5-elemene 1,4-dien-6-one
13-elemene
(3-farnesene Phenyl ethers
a.-guaiene
a.-humulene eugenol
isocaryophhyllene methyl eugenol
Y-murolene myristic in
a.-santalene safrole
a-selinene
13-selinene Oxygenated sesquiterpenes

5,10(15)-cadinen-4-ol
Oxygenated monoterpenes caryophylla-3(12),7(15)-dien-4-13-oJ
caryophylla-2,7(15)-dien-4-13-ol
borneol caryophylla-2,7(15)-dien-4-ol
camphor 13-caryophyllene alcohol
cavacrol caryophyllene ketone
caryophyllene oxide·
epoxy-dihydrocaryophyllene
nerolidol
4,10,10-trimethyl-7-methylene-
bicyclo-(6.2.0) decane-4-carboxal-
dehyde
 42
TABLE 5. CONTINUED

Hisc. compounds

butyric acid
3-methylbutyric acid
hexanoic acid
2-methyl pentanoic acid
benzoic acid
phenylacetic acid
cinnamic acid
piperonic acid
methyl heptanoate
methyl octanoate
2-undecanone
piperonal
m-methyl acetophenone
p-methylacctophenone
n-but.yrophenone
n-nonane
n-tridecane
n-nonadecane
piperidine
(methyl esters also found)
TABLE 6. MONOTERPENE HYDROCARBONS OF VOLATILE OIL FROM NAMED CULTIVARs78

(INTEGRATED AREA PERCENTAGES)

·hellan-
Black pepper 'ne and
samples n-Thu.jene a-Pinene Camphene Sabinene 13-Pinene Myrcene n-Phellan- 5-3- n-Terpi- p-cy1nen .imo- 'f-rerpi- Terpino- Total
drene Carene nene tene nene lene

Arikottanadan 1. 73 16.20 0.79 5.74 10.19 2.05 0.69 0.06 1. 26 0.13 .7. 63 1. 63 0.63 53.93

Balancotta 0.66 13.53 1.11 O.H 9.33 2.13 7.60 7.53 1. 61 1. 79 9.-!5 1.42 1.13 67.83

Cherivakanayakadan 3.07 6.67 0.32 8.32 7.38 2.02 2. 94 8.47 2.41 0.69 .6. 41 3.47 1. 55 64.22

Karimunda 0.25 6.12 0.45 0.39 11.20 2.53 3.36 15.52 0.58 0.46 8.37 0. 52 1.12 60.87

Karimkotta 0.39 5.72 0.30 0.39 8.48 1. 92 14.52 9.04 0.92 1. 84 .3. 93 0.34 0.70 63.49

Kaluvally 0.47 10.91 1.44 0.14 9.22 2.30 13.25 5.17 2.32 1.15 4.36 1.11 1. 21 73.09

Kuthiravally 3.27 5.40 0.24 13.73 8.30 1.84 1.17 0.46 1. 75 0.25 .8.61 2.52 0.90 58.49

l1unda 1. 38 1.11 0.30 3.59 4.92 2.08 2.88 6.73 2.49 0.38 .8.18 3.31 1. 5-! 49.19

Narayakodi 1. 53 5.91 0.23 7.46 11.59 2.01 0.46 0.03 0.73 0.23 .9. 79 1. 09 0.48 51.54

Nilgiris 2.18 10.44 0.49 5.47 7.11 1.66 11.07 0.26 1. 99 0.91 :I. 59 2.2-l 0.84 66.25

Panniyur-I 1. 62 6.99 0.34 4.81 14.33 1.99 0.83 0.33 0.82 0.17 :I. 30 1. 04 0.50 55.07

Sullia 0.57 10.67 0.43 0.28 7.99 1.67 27.37 0.21 1.42 2.78 3.61 0.25 0.28 77.53

Black pepper oil 2.15 4.88 0.26 9.45 7.97 1.43 1.10 4.36 1. 61 1.41 :o. 71 2.24 1. 01 58.58

steam distilled
TABLE 6. CONTINUED

SESQUITERPENE HYDROCARBONS AND OXYGENATED COMPOUNDS OF VOLATILE OIL FROM NA~ CULTIVARs78

(INTEGRATED AREA PERCENTAGES)

Total
sesqu-
Cultivars a-Cube- a-Cop- 0-Ele- B-Carvo- 0-Farn- Humu- B-Se1i- a-Se1i- 0-Biso- iterpene tina- rp- a-Terp- Carv- Caryo- Total
bene aene mene phyllene esene lene nene nene bolene hydro- 1oo1 inol one phyllene oxvge-
carbons 1 oxide nated
compo-
unds

A1·ikot tana<.lan 0.76 0.36 O.i2 18.79 2.30 1.45 0.58 2.08 2.34 29.38 0.16 0.03 o.o 0.04 5.9

Balancotta 0. 21 0.16 1.12 22.5-1 0.24 1.40 1. 57 1.43 0.21 28.93 0.07 0.01 0.01 0.04 2.1

Ched yakanayakadan 1. 1 i 3.35 0.81 13.49 0.23 1. 22 1. 55 1. 3-! 0.09 28.75 o. 20 0.03 o.o1 o.o-t 3.9

Karimunda 0.-13 0.16 2.43 21.50 0.56 1. 99 4.61 3.34 0.24 35.32 0.20 O.Oi o.o 0.21 2.0

Karimkotta 1.12 0.05 1. 06 25.61 0.33 1. 52 1.17 0.95 0.09 31.90 0.18 o. 01 0.04 3.1

Ka1uvally 0.69 0.24 0.96 12.63 0.40 1. 02 1.41 1. 21 1. 74 20.30 0.0-t 0.06 0.05 3.2

Kuthiravally 0.45 0.16 0.66 9.39 2.23 2.14 1. 01 3.07 3.53 22.64 0.20 0.03 0.0 0.06 15.7

Munda 1.6-1 0.06 0.87 29.8-t 3.26 1. 59 0.40 2.43 1.86 41.95 0.11 0.02 o.o 0.43 5.4

Narayakodi l.H 0.10 2.05 30.94 I. 04 2.02 2.19 1. 56 O.H 41.18 0.21 c 0.05 0.01 0.04 5.5

Nilgiris 0.9i 0.10 1.20 20.53 1.88 1.31 1. 70 I. 27 0.17 29.13 0.05 0.09 0.01 0.17 3.0

Panni}~r-I 0.70 3.62 0.36 22.34 0.0-t 1.14 0.33 o.3o 5.18 34.01 0.25 0 0.03 0.0 0.09 6.5

Sullia 0.44 0.11 1.21 12.33 0.03 0.96 1. 56 1. 67 0.20 18.52 0.13 0 o.os 0.04 o.os 2.5

Black pepper oil 0.32 0.45 0.60 19.06 2.10 1. 23 I. 31 0.93 2.80 28.85 0.49 0 0.09 0.05 1. 30 7.3

steam distilled
 45

maturation period. Buckle et a]61 studied the composition of

green, black and white peppers of Sri Lankan cultivars (Table 7)
at various stages of maturity and reported a variation in the
content of volatile oil constituents between the three types of
pepper products at different maturity stages. A recent report82
on the composition of Indian green peppers (dried) at different
maturity stages did not reveal any definite pattern of change in
chemical constituents during development (Table 8). A higher
concentration of ~-pinene and limonene and a lower content of ~­

caryophyllene could be noted in these oils compared to that of

Sri Lankan varieties.

Determination of relative abundance of sabinene, ~-pinene, o-3-

carene, limonene and cryophyllene in steam distilled pepper oils
could be used as an index to differentiate geographical sources
of black pepper81, Analysis of a few Indian pepper cultivars of
different grades82 are presented in Table 9. It appears from the
table that the extractives, piperine and volatile oil content
varies with the grade.

Sensory analysis techniques have been used by various

workers64,74,77,83 to evaluate the odour quality and to identify
the compounds responsible for the typical aroma note of pepper.
While some of these workers found the typical peppery note in
volatiles of black pepper was associated with monoterpene
hydrocarbons74,83, others found the high boiling fraction as
having a true pepper aroma 77. Howeve.r, no single compound was
isolated that was peppery in aroma.

1.8.3.2 Pungency

Oersted84 in 1820 isolated from the extract of black pepper a

yellow crystalline substance called piperine which he found to
have a pungent taste. It was later identified chemically as
trans-trans form of 1-piperoyl piperidine85, Few other minor
alkaloids with some degree of pungency have been isolated from
 46

TABLE 7. COMPOSITION OF SRI LANKAN CULTIVAR PEPPER OIL (% V/V)67

Age of berries (months)

2 3 4 5 6 4 4.5 5 5.5 5

Constituent Black pepper Green pepper White pepper

Munoterpene hydrocarbons

a-Pinene 9,2 9.3 9.3 10.3 10.8 9.5 9.5 9.4 10.3 9.5 10.1 10.
Camphene 0.6 0.6 0.6 0.6 0.6 0.6 0.6 0,5 0,5 0.6 0.6 o.
13-Pinene 9.2 7.1 7.1 7.3 7.2 9.1 9.2 9.4 7.3 8.4 8.2 7.
Sabinene 6.8 6.5 6.5 7.0 5.4 5.9 6.8 6.5 6.3 5.8 5,4 5.
a-Phellandrene 5.0 4.8 4.8 4.0 3.6 4.8 3.7 4,0 3,5 4.1 3.8 3.
Myrcene 3.0 3.0 3.0 3.6 2.4 3.0 2.4 2.4 3.0 3.6 2.8 2.
a-Terpinene 1.•1 0.9 0.9 1.1 1.2 1.0 1,"1 1.2 1.5 0.8 1.1 1.
Limonene 8.7 9.9 9.9 12.2 12.6 9.5 9.7 9.9 10.9 10.2 10.9 12.
r-Terpinene 1.2 1.0 1.0 1.0 1.2 1.2 1.4 0.6 0.6 1.0 1.1 1.
p -Cymene tr* 0.3 0.3 tr tr 0.3 tr tr tr 0.3 tr tr
Terpinolene 0.8 0.6 0.6 0.4 0.5 0.6 0.6 0.6 0.8 0.5 0.6 o.
Total monoterpenes 44.9 43.9 44.0 47.6 45.5 45.5 45,0 44.5 44.7 44.8 44.6 44.

Sesquiterpene hydrocarbons

a-Cube bene tr tr tr tr tr 0.3 tr 0.3 0.3 0.6 ·o.6 o.

o-Elemene 2.3 1.8 2.2 1.9 1.8 2.3 2.0 1.9 1.8 1.9 1.9 1.
a-Copaene 8,9 8.5 7.3 6.1 6,0 7.2 7.3 5.8 5.6 7.2 6.3 5 ,I
13-Caryophyllene 17.7 19.7 17.6 23.6 26.0 17.6 18.9 22.2 23.2 21.2 22.2 23.
Humulene 0.5 0.5 0.4 0.4 0.5 0,6 0.6 0.7 0.7 0.6 0.7 o.'
13-Farnesene 5.4 6.0 5.4 2.4 2.4 6.0 5.4 4.5 2.4 3.9 3.9 2.
Calamenene 0.6 0.6 0.6 0.6 0.6 0.9 1.2 0.7 0.7 0.6 0.7 o.'
Total sesquiterpenes 35.4 37.1 33.5 35.0 37.3 34.9 35.4 36.1 34.7 36.0 36.3 34.

Oxygenated compounds

~-Terpinen-4-ol 1.2 1.3 1.3 o.8 o.8 1.2 1.6 1.0 1.3 1.8 1.8 I.:
Pinocarveol 1.2 1.8 2.3 0.9 0.9 1.8 1.1 2.1 1.1. 1.0 0.9 1.•
Myrtanol tr tr tr tr tr 0.6 2.1 0.8 0.9 tr 0.6 O.l
Caryophyl1ene oxide 2.6 3.8 4.8 3.2 3.2 3,0 3.2 2.9 3.0 3.8 2.9 3,:
Humulene oxide 0.6 tr tr tr 0.6 0.6 0,8 0.8 0.7 0.6 0.7 0.!
Total oxygenated campo- 5.6 6.9 8.4 4.9 5.5 7.7 8.8 7,6 7.0 7.2 6.9 7 ••
unds
Unknown fraction 14.1 12.1 14.1 12.5 11.7 11.9 10.8 10.8 13.6 12.0 12.2 12.!
Total of all- 85.9 87.9 85.9 87.5 88.3 88.1 89.2 88.2 86.4 88.0 87.8 87,<
constituents

*tr Trace
TABLE 8. GC ANALYSIS OF PEPPER OILS AT DIFFERENT MATURITY WES
(DRIED)82

Sl. Constituents 1 month 2 months 3 months .4 months 5 months 6 months months

No.

01. a.-Pinene 2.17 5.50 6.74 7. 74 7.85 6.85 6.43

02. Camphene O.Oi 0.14 0.1i 0.21 0.20 0.18 0.17

03. 13-Pinene 6.83 22.13 29.43 29.63 31.76 30.00 30.07

04. G-3-Carene 0.05 0.27 0.36 0.45 0.46 0.40 0.22

05. d-Limonene 7.58 14.89 18.53 1i. 71 21.37 22.46 23.40

06. Lina1ool 0.81 o. 54 0.34 0.60 0.48 0.50 o. 51

07. Terpenen-4-ol 0.22 0.75 0.62 2.28 I. 04 0.66 0.57

08. a.-Terpenic! 0.18 0.22 0.21 0.40 0.20 0.19 0.21

09. Carvone 0.38 0.65 o. 50 0.48 0.38 0.35 0.35

10. Piperitone 0.45 0.25 0.16 0.15 0.13 0.13 0.15

11. Caryophyllene 7.31 9.99 10.43 8.94 8.40 9.70 9.54

12. Humulene 0.91 0.76 o. 74 0.67 0.61 0.66 0.66

13. Cadinene 10.76 10.69 9.20 8.58 7.22 8.00 5.64

TABLE 9. ANALYSIS OF GRADES OBTAINED FROM DIFFERENT PEPPER VARIETIEs82

Volatile oil
Variety Grade Extrac- Pipe-
tives rine % % yield *

Karimunda (PH Spec.) 2.3 0.6 0.9

GL-Special 8.0 5.4 3.0 80
MG-1 7.6 4.8 2.4 80
TG 6.5 3.9 2.2 78
TGSEB 6.9 3.8 1.3 82

Kottanadan (PH Spec.) 5.7 2.2 1.0

GL-Special 10.9 5.3 3.0 90
MG-1 7.6 3.9 3.3 77
TG 7.0 4.0 2.7 80
TGSEB 5.8 3.6 2.2 80

Kalluvally (PH Spec.) 3.5 1.1 0.9

GL-Special 8.9 5.1 2.7 87
MG-1 8.7 5.1 2.9 82
TG 8.3 4.6 3.4 83
TGSEB 7.6 3.9 3.0 82

Panniyur (PH Spec.) 3.5 0.8 1.0

GL-Special 10.1 5.4 3.0 89
MG-1 8.8 4.8 3.1 86
TG 8.9 4.6 3.0 85
TGSEB 8.0 4.3 3.0 83

~
CP
*RI = Refractive index
black pepper86, 87' 88, They are piperettine, piperyline,
piperanine and piperolein A and B. Spectrometric methods56,86,89
based on the absorption by piperine in the UV range at 345 nm was
found to be a simple, rapid and widely used method for
quantitative estimation of piperine. TLC coupled with
spectrophotometric methods have also been used to give a better
estimation of pungency64, 90. GLC91-93 and HPLc94' 95 have been
recently used to estimate piperine and other minor pungent
principles. Sensory techniques using seaville heat unit has also
been used to evaluate the pungency in pepper64,88, A clear
agreement between sensory pungency and that estimated
spectrometrically at 342 nm was observed64,

Piperine content of black pepper is generally found to be

2-6% 96,97, Black pepper oleoresin contains 35-40% piperine as
estimated by Spectrophotometric method98. Piperine content was
recorded to be 40-58% in C02 extract99. Green pepper products
prepared from immature berries contain higher piperine content
than matured ones as piperine content reaches a maximum. before
maturi ty63 '82. Removal of outer pericarp during processing
results in high concentration of piperine in white pepper64,
Piperine content of four varieties of Indian pepper is depicted
in Table 9.

1.8.3.3 Phenolic constituents

The phenolic constituents of pepper have received much less

attention in spite of their organoleptic significance as
explained earlier. Schulz and Herramann45 isolated and identified
nine phenolic acids namely caffeic, ferulic, gentisic, p-
hydroxybenzoic, protocatechuic, salicylic, synapic, syringic and
vanillic acids in black pepper using GLC after prior
derivatization into their silylated derivative. These phenolic
acids were shown to exist solely as their glycosides in pepper
and exhibited qualitative and quantitative differences in their
 50

distribution in black and white pepper45. Among the phenoli

acids reported, syr ing ic acid was found to be the predomi_nan
phenolic acid of black pepper, while ferulic acid dominated i
white pepper45. White pepper with a total phenolic acid content
1/5th that of black pepper(673 mg/kg) was reported to be devoi
of caffeic, synapic, gentisic and salicylic acids45.

Vosgen and HerrmannlOO reported the presence of considerabl

concentration of glycosides of kaempferol ( 1, 500 mg), rhamneti
(600 mg) and quercetin (350 mg) with lesser amounts o
isorhamnetln glycoside (<20 mg) per kg of black pepper usin
paper chromatography and HPLC. Glucose, rhamnose, a~abinose an'
rutinose were identified as the major sugar residueslOO,

Nakatani et al 101 reported the presence of two phenolic amide

namely N-trans feruloyl tyramine and N-5-(4-hydroxy phenyl)-2E,4
-pentadienoyl piperidine (coumaperine) in yield of 325 mg and 13.
mg per kg of lvhite pepper respectively. Three new phenoli,
compounds namely N-trans-feruloyl piperidine, N-5- ( 4-hydroxy-3·
methoxy phenyl)-2E,4E-pentadienoyl piperidine (feruperine) and N·
5- ( 4-hydroxy-3-methoxy phenyl) -2E-pentenoyl piperidine were alsc
isolated and characterized in white pepper by spectroscopic
analysis! 02. They were obtained in yields of 11 mg, 17 mg anc
9. 5 mg per kg of white pepper respectively102. Coumaperine anc
feruperine were suggested to play a crucial role in the
biosynthetic pathway_of piperine102,

No report exists on the nature of phenolic compounds in gree1

pepper except for a report on the polyphenol content at differen1
stages of maturity before and after processing82. Pepper beforE
drying showed the highest amount of polyphenol content (1%) whicl
reduced to 0.4% on ripening82.
 51
1.9 CINNAMON
Z.EYLA~:CCU.M" OR C:C~.N"A.M"O.M"U.M"

V.E.RU.M")

The dried bark of Cinnamomum verum constitutes the true cinnamo;

of commerce. Sri Lanka leads in the production and export o
cinnamon. Seychelles &nd Malagasy republic are the other tw'
countries with a major share in cinnamon trade. Cassia
considered to be inferior to true cinnamon are grown mainly L
China (Cinnamomum cassia). Most of the demand for cinnamon i
India is met by imports.

C. verum or C.zeylanicum is an evergreen tree that grows to .

height of 8-17 rn and 30-60 ern in diameter. It has thick gra
bark and low lying branches with elliptic dark green leaves. Th1
bark and leaves are strongly aromatic. The yield of cinnarno.
bark is generally 168-224 kg of quills per hectare annually 63 k1
of chips and 2. 5 tonnes of undried· leaves per hectare are als'
obtained annually.

1.9.1 Products and uses

The products14 of cinnamon that are available in the market ar1

the whole/ground bark, its oleoresin and the essential oils.

Cinnamon bark has a warm spicy, aromatic and woody aroma. Whol1
or ground cinnamon bark is used mainly for culinary purposes an1
for flavouring of bakery products, sauces, pickles, puddings
curry powder, beverages and confectionery.

Cinnamomum zeylanicum yields 10-12% of oleoresin which contain:

16-60% volatile oil. Oleoresins are prepared mainly by flavourinl
industries and have similar application to ground spice.

Cinnamon bark essential oil is a pale yellow liquid possessin1

the delicate aroma of the spice and a sweet pungent taste·. Barl
 52

oils are used to flavour baked goods, sauces, pickles,

confectionery and beverages. They are also used in perfumery and
in dental and pharmaceutical preparations. Leaf oil is used in
flavourings and perfumery.

1.9.2 Medicinal properties

Cinnamon is used8,66 as a carminative, astringent, stimulant and

antiseptic. It stops vomiting, relieves flatulence and is useful
for diarrhea and hemorrhage.

1. 9. 3 Chemistry

The dried bark of cinnamomum verum contains essential oil, fixed

oil, tannins, resins, proteins, cellulose, pentosans, mucilage,
starch, calcium oxalate and mineral elements14,

Organoleptic properties of cinnamons are determined by the

composition of their essential oils. The polyphenols that make
up the bark tannins also play a role in contributing t·o the
colour of the spice.

1.9.3.1 Cassia

The general composition of Cinnamomum cassia is similar to that

of C. verum except for a higher mucilage content. The essential
oils of cassia bark and leaf have been extensively studied by
several. workers103' 104. C. cassia differs from c. verum in that
only one type of essential oil is obtained from both stern bark
and leaves. Cinnamaldehyde is the major constituent in both
oils. Another distinguishing feature is the presence of o-
• methoxy cinnamaldehyde and coumarin in cassia bark oils in
contrast to C.verum where they are absent. Eugenol on the other
hand is present only in c.verum.
 53

1.9.3.2 Essential oil

Cinnamomum verwn plant yields three types of volatile essentia:

oils namely the bark oil, the leaf oil and the root oil. 0:
these the bark oil is the most important while the root oil ha:
no commercial significance. The chemistry and technology o
e s s. e n t i a 1 o i 1 of c i n n am o n p 1 ant has be e n r e v i ewe d b:
Wijesekara105,

(a) Bark oil

The yield of oil obtained from bark ranges from 0.5 to 2:

depending on its origin. The oil content of Indian cinnamon8:
bark range from 2-2.2%.

Earlier work on the chemistry of cinnamon bark and leaf oil ha:
been well documented in literature where the abundance o:
cinnamaldehyde in bark oil and eugenol in leaf oil wa:
reported106. With the help of chromatography and spectrometri<
techniques, Bramaramba and Sidhu107 identified 13-caryophyllene.
benzyl benzoate, o-methyl eugenol, p-cymene, linalool, a-pinene
g-phellandrene and humelene in the non phenolic fraction of barl
oil of Cinnamomum ze.vlanicum grown in India. l'Hjesekara et a]lOl
reported a detailed gas chromatographic examination of bark, roo1
bark and leaf oils of Sri Lankan cinnamon. Twenty-six compound:
were identified in the bark oil in this study. Senanayake ei
a]109 have analyzed the bark, leaf and root bark oils oj
C.zeylanicum. A total of 72 compounds were identified in all thE
three oils of which 11 monoterpenes, 4 sesquiterpenes, onE
aromatic and two aliphatic compounds were reported for the firs1
time in these oils. Table 10 lists the constituents identifiec
by these workers. A recent report82 on the composition oj
different varieties of Indian cinnamon bark and leaf oils alsc
confirmed the presence of the above constituents.

Cinnarnaldehyde constituted 65-75% of the Sri Lankan14 and 40-55~

of the Indian82 bark oils while Seychelles and Malaga~y bar~
 54

TABLE 10. VOLATILE CONSTITUENTS IDENTIFIED IN COMMERCIAL CINNAMON

( CLNNAMOMUM ZEYLANICUM) LEAF, STEM BARK, AND ROOT BARK
OII.S OF 5-YEAH Ol.D PLANT109

Peak Relative %

Retention Stem Root

No. time, min Compoound Leaf oil Bark oil Bark oil Identified by

1 9.0 a-Pinene 1.0 0.6 4.3 RD, PE, IR

2 9.4 Unknown Tr Tr 0.1
3 9.8 Camphene 0.4 0.2 1.9 RD, PE, IR
4 10.7 13-Pinene 0.3 0.1 2.2 RD, PE, IR
5t 11.7 Sabinene Tr 0.02 0.8 RD, PE
6t 11.8 ~ -3-Carene 0.1 0.03 · Tr RD, PE
7 12.0 Myrcene 0.1 0,1 2.2 RD, PE
8 12.3 a-Phe1landrene 0.1 0.6 0.2 RD, PE, IR
9 12.7 a-Terpinene 0.1 0.4 0.2 RD, PE, IR
10 13.3 Limonene 0.4 0.5 3.1 RD, PE, IR
11 13.7 1:8-Cineo1e 0.6 2.0 11.7 RD, PE, IR
12*t 14.3 cis-Ocimene 0.02 0.03 0.6 RD, PE
13t 15.0 1-rerpincne 0.1 0.03 0.2 RD, PE
14*t 15.1 trans-Ocimene 0.02 Tr 0.2 RD, PE
15 15.9 p-Cymene 1.2 1.1 1.4 RD, PE, IR
16"' 16.4 Terpinolene 0.1 0.1 0.3 RD, PE, IR
17*t 18.3 11-Hexanol Tr Tr ND RD, PE
15*t 21.6 Fenchone Tr Tr 0.02 RD, PE
19t 24.3 Furfural 0.02 0.03 0.02 RD, PE
2o*t 25.0 trans-Lina1ool oxide 0.1 0.1 0.1 RD, PE
furanoid form
21 26.8 a-Ylangene 1.0 0.3 ND RD, PE, IR
22*t 27.0 Nonana1 Tr ND ND RD, PE
23* 27.4 Benzaldehyde 0,7 0.3 ND RD, PE, IR
24 27.8 Camphor Tr Tr 56.2 RD, PE, IR
25 28.7 Lina1ool 3.4 2.4 0.9 RD, PE, IR
26*t 29.4 Linalool acetate Tr Tr 0.3 RD, PE
27*t 30.7 Borny1 acetate Tr 0.1 0.03 RD, PE
28 31.7 Terpinen-4-ol 0.1 0.4 2.7 RD, PE, IR
29 32.3 Caryophy Liene 5.8 3.3 0,5 RD, PE, IR
3o*t 33.4 2-Phenylacetaldehyde Tr Tr 0.04 RD, PE
Jl*t 34.6 Borneol Tr 0.02 0.1 RD, PE
32* 35.0 Methyl chavicol Tr Tr 0.4 RD, PE, IR
33 35.8 a-llumulene 0.9 0,6 0.2 RD, PE, IR
34 36,4 «-Terpineol 0.4 0.7 6.9 RD, PE, IR
3s*t 37.2 13-Selanene Tr Tr ND RD
36*t 37.9 Y-cadenene 0.01 ND 0.5 RD, PE
37*t 38.1 Geranial 0.02 Tr ND RD
38* 38.5 Piperitone 0.04 0.1 0.9 RD, PE, IR
39t 38.7 Geranyl acetate ND Tr ND RD, PE
40* 39.0 Cumina1dehyde 0.1 0.04 0.1 RD, PE
 55

TABLE 10. CONTINUED

Peak Relative :1;

Retention Stem Root

No. time, min Compoound Leaf oil Bark oil Bark oil Identified by

41* 39.6 Hydrocinnamaldehyde 0.2 0.4 Tr RD, PE, IR

42*t 40.3 Phenylethyl acetate 0.03 0.1 Tr RD, PE
43t 41.4 Nero! Tr Tr ND RD
44 44.1 Geraniol 0.04 0.1 0.7 RD, PE, IR
45'kt 44.8 Benzyl alcohol Tr Tr Tr RD, PE
46 45.7 Safrole 2.3 Tr 0.3 RD, PE, IR
47"' 46.8 2-Phenyiethyl alcohol Tr 0.4 Tr RD, PE, IR
48 48.1 Unknmm ND ND Tr
49* 49.2 2-Phenylpropyl acetate 1.0 0.1 Tr RD, PE, IR
50 50.4 Unko'h'n Tr Tr ND
Sl'tt 51.8 Phenol 0.02 Tr Tr RD, PE
52* 52.5 Caryophyllene oxide o.s Tr 0.2 RD, PE, IR
53*t 52.9 Methyleugenol 0.01 Tr ND RD, PE
54 53.5 Cinnamaidehyde 2.0 75.0 0.7 RD, PE, IR
sst 54.5 Methyl cinnamate 0.03 Tr Tr RD, PE
56 55.1 Unknown Tr ND Tr
57*t 56.3 11ethy I i soeugeno I Tr ND Tr RD, PE
58 56.9 Unkno ..·n ND ND ND
sst 58.8 Ethyl cinnamate 0.02 ND Tr RD, PE
60 60.1 Cinnamyi acetate 1.7 5.0 Tr RD, PE, IR
61 60.6 Eugenol 70.1 2.2 0.5 RD, PE, IR
62 62.9 Unknown Tr ND ND
63 63.8 Acetyleugenol 2.5 0.2 0.1 RD, PE, IR
64 64.5 Unknown Tr Tr ND
65 65.2 Cinnamyl alcohol 0.4 0.3 0.1 RD, PE, IR
66*t 67.1 Farnesoi 0.1 0.03 ND RD, PE
67t 68.8 Isoeugenol 0.1 0.02 ND RD, PE
68*t 69.2 2-Vinylphenol Tr 0.03 ND RD
69* 71.3 Coumarin Tr 0.7 ND RD, PE, IR
7o*t 72.8 Vanillin ND Tr ND RD, PE
71 79.• 2 Denzylbenzoate 3.5 0.7 0.3 RD., PE, IR
n*t 83.1 2-Phenylethyl benzoate Tr ND ND RD

ND, not detected; *, new compound; t, tentative identification; RD, retention data; PE, peak
enhancement; IR, infrared spectrum

•
@

56

oils14 showed a cinnamaldehyde content of 62-65% Eugenol

content, on the other hand, was found to be 5-18% in Sri Lankan
oils with Seychelles and Malagasy oils registering a content of
2-10%. Eugenol was, however, found to be low (0.5-1.3%) in Indian
bark oils82. Lower amounts of mono terpene hydrocarbons and
linalool (1.6-1.8%) with a higher content of cinnamyl alcohol (1
to 3.3%), cinnamyl acetate (30 to 35%) and benzyl benzoate (1 to
2.5%) were also noted in these oils82 compared to the steam
distilled oils from Sri Lankan cinnamon14 (linalool 6.23%,
cinnamyl alcohol 0.43%, cinnamyl acetate 5% and benzyl benzoate
0.21%).

(b) Leaf oil

Cinnamon leaf oil is a yellow to brownish yellow coloured liquid

that possesses a warm, spicy and harsh aroma lacking the rich
body of cinnamon bark oil. The yield of oil varies from 0. 5 to
0.9% • In contrast to the bark oil, the leaf oil contains
eugenol as the major constituent. Table 10 lists the
constituents identified in cinnamon leaf oil.

While Sri Lankan oils showed a eugenol content ranging from 60-
90%, in Seychelles and Malagasy cinnamon it ranged from 70-95% •

Among the Indian leaf oils studied, the Calicut cinnamon had an
eugenol content of 90% with Trichur and Mangalore cinnamons
showing 78.6% and 52.66% respectively82. A distinguishing
feature of Indian cinnamon leaf oils grown in Bhubaneswar is the
relatively high benzyl benzoate content110. Among the other minor
constituents identified in Indian cinnamon leaf oils, ~­

caryophyllene (2.77%) safrole (0.5-1%) and linalool (0.88%)

were in detectable amounts in Calicut oils, while eugenol acetate
(3.77%), benzyl benzoate (2.74%), 13-caryophyllene (2.19%),
linalool (2.11%) and a-ylangene (2.14%) were the important minor
constituents of Trichur cinnamon leaf oils82, Indian cinnamon
variety from Mangalore, however, had a leaf oil containing
 57

eugenol acetate (6.9%), methyl eugenol (2.5%) and a-caryophyllenE

(7.08%) in relatively high proportions82,

(c) Root bark oil

It is a colourless pale yellowish liquid with

brown
camphoraceous odour. Yield varies from 1-2.8% . Major component
of the oil is camphor constituting 60% of the oil14.
Constituents identified in root bark oil are included in Table
10.

1.9.3.3 Phenolic constituents

Schulz and Herrmann45 have reported the presence of ten phenolic

acids totalling to 414 mg/kg in Sri Lankan cinnamon bark of which
protocatechuic a~id was the prominent compound. Presence of
epicatechin, gallocatechin, epigallocatechin and proanthocyanins
such as cyanidin have also been reported by them111 in Sri Lankan
cinnamon bark. Nonaka et a1112 have isolated and identified
trimeric, tetrameric and pentameric proanthocyanidins from
Cinnamomum zeylanicum using standard chemical and spectral
methods. Tannins of Chinese cinnamon ( Cinnamomum cassia) were
also studied by Marimoto et al113, 114, while Buchalter115 found
that the tannins of Saigon cinnamon (Cinnamomum loureirii)
consists of polymeric units of 5,7,3',4'-tetrahydroxyflavan-3,4-
diols.

1.9.3.4 Sensory evaluation

Subjective-objective evaluation has been used recently in the

identification of aroma character impact components in cinnamon
bark and leaf, where the oxygenated aromatic compounds were shown
to be important116,117,
 58

1 . 10 CLOVE ( SY..ZYGIUM" .AROM".ATICUM"

OR .EUGEN"I.A C.ARYO.P.EIYLLUS)

Unopened flower buds of Syzigium aromaticum constitute the cloves

of commerce. Although indigenous to Moluccas, it is mainly
produced in Zanzibar and Halagasy republic (Madagascar).
Indonesia is the world's largest consumer of cloves. India
imports clove to a large extent with small amounts being produced
internally.

Clove tree is a small ever green tree reaching a height of 12-15

rn and 30 em in diameter. Leaves are aromatic. Several cultivars
of clove are grown. Dry clove buds weighs 1/3rd the weight of
green freshly harvested cloves. The mean average yield of green
clove is around 1375 kg/hectare per annum. Weight of dry stem
produced is 1/3rd that of green stem and about 23.5% of dry
cloves.

1.10.1 Products and uses

Commercially, clove is available in three main forms namely (i)

whole clove buds used for preparing ground spice, oleoresin and
bud oil (ii) stem oil and (iii) leaf oil.

Traditionally dried clove buds are obtained by sun drying of

green clove buds. Clove buds in the whole/ground form is used
for domestic culinary purposes, sauces and pickles and for
flavouring of kretek cigarettes in Indonesia. Oleoresins are
prepared using a variety of solvents. Oleoresin is used for
flavouring manufactured foods and in perfumery. The essential
oils obtained from bud, stern and leaves of clove are also used in
flavouring, perfumery and pharmaceutical industry. ·

1.10.2 Medicinal properties.

Clove8' G6 is the most stimulating and carminative of all

j
 59

aromatics and is given in powder or infusion for nausea, emesis,

flatulence, languid indigestion and dyspepsia. It stimulates
peristalisis, is a strong germicide, a powerful antiseptic and a
local anesthetic applied to decayed teeth.

1.10.3 Chemistry

Dried clove buds contain steam volatile oil, fixed oil,

phenolics, res ins, tannins, proteins, cellulose, pentosans and
mineral elements. Carbohydrate comprises 2/3rd of the weight of
the spice. The organoleptic properties of cloves are determined
by the composition of their essential oil.

1.10.3.1 Essential oils

The th~ee commercially important essential oils of clove are,

l.>ud, stem and leaf oils. Of these the bud oil is the most
important. All the three oils contain eugenol as the main
constituent. They, however, differ in their odour
characteristics.

(a) Bud oil

The steam distilled clove bud oil is a pale yellow to colourless

liquid possessing the odour and flavour characteristics of thE
spice. It is obtained in yields of 15-17%14. Use of
supercritical liquid C02 as extracting solvent has recently been
reported to yield 19% of volatile oil from clove buds118.
Variation in the conditions of supercritical C02 extraction,
however, resulted in higher yields of oil ranging from 21-24%57,

Analysis of essential oil of clove bud was carried out as early

as 1875 by Churchll9 who reported the presenc~ of humelene
besides eugenol as one of the constituents of this oil. Studies
undertaken between late 19th century and early 20th century
resulted in identification of 18 components in the clove bud
oil14, A number of GC studies on bud oil conducted by later
 60

workers 14 resulted in the addition of 8 more compounds to this

list; the major constituents identified in the oil were eugenol,
eugenol acetate and ~-caryophyllene. Trace components present in
the oil included chavicol, methylsalicylate and vanillin among
the phenols along with terpene hydrocarbons and various
oxygenated, aliphatic as well as aromatic compounds14, A recent
report82 on the composition of clove bud, stalk and leaf oils
from Indian clove plant gives a list of 31 compounds identified
in all the three oils which is reproduced in Table 11. Zheng
et a1120 have very recently reported the presence of a-humelene
epoxide among the sesquiterpenes studied ~n clove bud oil.·

Eugenol is the major constituent generally accounting for 70-90%

of the bud oil with eugenol acetate and ~-caryophyllene being
present to the extent of 17% and 5-12% respectively14. The bud
oil constituted mainly of oxygenated compounds contributing to
90% of the oil Hi th mono terpene and sesqui terpene hydrocarbons
accounting for 2% and 8% of the oil, respectively. Conditions
used for processing such as heat treatment and storage can have a
pronounced effect on the composition of volatile oils121,

(b) Stem oil

It is a pale yellow liquid possessing a coarser and more woody

odour than the bud oil. Higher eugenol content (83-95%) together
with lower amounts of eugenol acetate compared to bud oil is
responsible for its harsher phenolic odour. They contain all the
constituents detected in bud oil82 (Table 11).

(c) Leaf oil

It is obtained in yields of 2-3% as a dark brown liquid

possessing a harsh, woody and dry phenolic odour. It has a lower
eugenol content (80-8~%) than bud oil. Leaf oil has similar
qualitative composition as bud oil82 (Table 11).
Gopalakrishnan et a1122 showed that the content and composition
of Indian clove leaf oil varies during maturation; of the major
 61

TABLE 11. COMPOSITION OF CLOVE BUD, S'l'EM AND LEAF OIL~.82

% Compc,sition

Compound RRt Bud Stem Leaf

a-Pinene 0.42 0.02 0. (13 0.03

13-Pinene 0.44 0.03 0. (J6 0.03

2-Hexanone* 0.48 0.05 0.05 0.03

2-Heptanone + 1:8-cineole 0.50 0.05 0.04 0.06

a-Terpinene + Limonene 0.53 0.02 0.(12 0.01

p-Cyrnene 0.56 0.05 0.05 0.05

2-Heptanol 0.06 0.03 0. () 1

2-Nonanol 0.64 0.05 0.06 0.03

u. I. 0.66 0.04

Benzaldehyde 0.69 0.07 0. 1)3 0.07

13-Terpineol*(tentative) 0.81 0.07 0.12 0.13

Terpinen-4-ol* 0.87 0.02

a-Cube bene* 0.90 0.66 0. ·19 0.08

a-Copaene* +a-Terpineol* 0.92 0.84 0. 77 0.37

+ Benzyl acetate

.1.3-Caryophyllene 1. 00 6.18 6. "57 6.42

a-Humulene 1. 06 1.06 } }
} 1. 53 } 1. 39
Benzyl alcohol 1.10 0.52 } }

¥-cadinene* 1.14 0.34 0. :15 0.55

5-Cadinene* 1.18 0.54 Traces

Caryophyllenol 1. 34 0.08

Eugenol (+ traces of 1. 41 77.13 8 0. t 9 80.97

sesquiterpene alcohols)

!so-eugenol (+ traces of 1. 51 0.42 0. 12 Traces

sesquiterpene alcohols)
 62
TABLE 11. CONTINUED

% Composition

Compound RRt Bud Stern Leaf

Eugenol acetate (+ traces of 1. 54 5.04 0.44 Traces

sesquiterpene alcohols)

u. I. 1. 57 0.71

Farnesol (c,t)* 1. 59 0.77 0.47 0.61

u. I. 1. 63 0.62 0.59

Farnesol (t,t)* 1. 66 1. 08 0.69 0.61

u. I. 1. 71 1. 28

u. I. 1. 72 0.22 0.51
Vanillin 1. 82 0.92 0.73
u. I. 1. 87 0.46 0.80

Dibutyl phthalate 1. 96 0.46 0.40 0.64

Asarone (tentative) 2.06 1. 08 0.50 ·o.45

u. I. 2.17 0.67 1. 32

" 2.32 0.37 0.33 0.29

" 2.44 0.44 0.28 0.55

" 2.52 0.07 0.28 0.55

" 2.54 0.42 0.37 0.15

" 2.77 0.02

" 2.80 0.01

" 2.81 0.03

RRt refers to relative retention time with re.spect to f3-

caryophyllene which is assigned an Rt of 1.0
* Refers to compound newly reported in the Gil
U.I. - Unidentified

J
 63

constituents, eugenol content increased from 38.2 to 95.2% while

eugenol acetate and 13-canrophyllene content decreased from 51.2
to 1.5% and from G.3 to 0.2%, respectively. Indonesian leaf oil
contained 13-caryophyllene as high as 18% with eugenol and
humelene content being 79% and 1.9%, respectively. Recently,
over 70 constituents were identified in this oil by GC/MS
studiesl23,

1.10.3.2 Phenolic constituents

Phenolic acid glycosides present in clove have been studied by

Schultz and Herrmann45, Of the ten phenolic acids identified
together accounting for 13.9 rug/kg, gallic acid was the
predominant followed by p-coumaric and protocatechuic acids
including other phenolic acids such as ferulic, caffeic,
gentisic, salicylic, p-hydroxy benzoic, vanillic and syringic
ac ids . V o s g en and He r r mann 1 0 0 have i dent i f i e d f 1 avon o i d
compounds such as kaempferol, quercetin and rhamnetin and their
glycosides in clove buds. Ellagl tanins which are gallic acid-
carbohydrate complexes have also been isolated from clove
budsl21, One of this, namely cugcnin \vas found to show antiviral
activityl25, 3,4-dihydroxy phenyl ethanol and 3,4-dihydroxy
benzoic acid were also isolated from leaves of clove plant and
shown to have anti-inflammatory propertiesl26,

1.10.3.3 Sensory evaluation

Salzer127 has suggested that eugenol, eugenol acetate and .13-

caryophyllene could be responsible for the t top aroma' note of
clove. Trace compounds present in the oils also contribute to
the organoleptic properties of the spice; for example methyl-n-
amyl ketone, a minor component is cons ide red to be responsible
for the fresh fruity top note14, The tannins present in clove as
mentioned above may pLay an important role in the appearance and
astrigcncy of dried clove. However, the relative importance of
various trace components in determining the odour character of
clove has not been clearly established.
 64

1 •. 1 1 CARDAMOM ( .EL.ETTAR.IA
CAR.DAMOMU.M')

Cardamom is principally the dried fruits of Elettaria cardamomt

(var. miniscule) also kno~n as small or true cardamom. India j

the largest producer and exporter of true cardamom with Guetarnal

being ranked second. International trade concentrates mainly c
small cardamom. Several varieties of small cardamom are grown
India of which the Malabar and Mysore varieties are the rna~

prominent.

They are perennial herbacjous plants growing to a height of 3 t

4 m with creeping root stalk and two distinct aerial growths U
leaf and flower shoots. Fruits are fleshy when green an
leathery when dry. Yield of dry fruit ranges from 25-lE
kg/hectare

1.11.1 Products and uses

Cardamom is mainly available in the market in three main formsl~

namely (i) in the whole as green and bleached (creamy or goldE
yellow colour) or ground form (ii) oleoresin and (iii) essentil
oil.

Whole cardamom is marketed as dry matured fruits. Fine quali 1

cardamoms shmv tightly packed, glistening dark brown seeds th~

are slightly sticky to touch. Whole and powdered cardamom a1

used by food and flavouring industries, in culinary preparation~

confectionery and sweets, in bakery and pastry products.

Cardamom essential oil j.s mainly used in food and perfurne1

industries and to a limited extent in liquor and pharmaceuticE
industries as a flavqurant and carminative. Due to the closene~

of the volatile oil and oleoresin in flavour quality oleoresir

are not of much commercial importance.
 65

1.11.2 Medicinal properties

Cardamom8,66 is a carminative and stimulant, used mainly as an

adjuvant or corrective. It is used in indigestion and flatulence
and is good for colic and disorders of the head. It cures skin
diseases, colds and inflammation.

1.11.3 Chemistry

The general chemical constituents of cardamom are the volatile

oil, phenolics, fixed oil, waxes, starch, proteins, fiber,
pentosans, vitamins, minerals and colouring matter. The content
of non volatile ether extract is reported to vary from 2-3.7%,
while the total ash content indicative of the cleanliness of the
product varies from 3.8-6.2% in seeds. The characteristic aroma
of cardamom is due to its essential oil128,

1.11.3.1 Essential oil

Cardamom essential oil is generally obtained in yields of 6-8% by

steam distillation and about 5.2% by cold pressing of freshly
decorticated seedsl29, Use of super critical ·coz extraction
under varying conditions of pressure, temperature and time
resulted in yield of oil ranging from 7.6% to 7.8%130, The yield
of oil varies both with maturity and the variety under
cultivation. Natarajan et aL131 observed an increase in volatile
oil content from 5.4-7.0% with advance in maturity. This was
also confirmed in a recent report by Kumara et a1132. Halabar
and Mysore varieties also showed considerable variations in their
oil content ranging from 6-12%131,

The early \V"Ork on the composition of steam distilled essential

oil of cardamom showed the presence of 1,8-cineol, d-u-terpineol,
terpinyl acetate, limonene, sabinene and borneol133, Using gas
chromatography, Ikeda et a173 identified monoterpene hydrocarbons
 68

such as a-pinene, a-thujene, ~-pinene, sabinene, a-terpinene,

myrcene, D-limonene, Y-terpinene and p-cymene in cardamom
essential oil. First detailed analysis of volatile oil of
cardamom was reported by Nigam et al134 who confirmed the
presence of above compounds and additionally identified methyl
heptanone, 1 inalool, 1 inalyl acetate, ~-terpineol, geraniol,
nerol, neryl acetate, nerolidol, 2-undecanone, 2-tridecanone and
heptacosane in the oil. A Japanese work on the analysis of
cardamom essential oils reported new compounds namely 1,4-cineol
and cis- and trans- p-menth-2-en-1-ol to be present in the
oil135. A systematic study on the composition of volatile oil
was reported by Lawrence136 who identified 55 compounds as
presented in Table 12, Maurer et a]137 isolated two unusual
hydrocarbons namely (E)-4,8-dimethyl-1,3,7-nonatriene and (E,E)-
4,8,12-trimethyl-1,3,7,11-tridecatetraene from essential oil of
cardamom. Pieribattesti et a]138 identified, besides the already
mentioned constituents, p-menthene, methyl allyl benzene, 1-
octanol, borneol, trans-carveol, hydroxy acetates, farnesol,
methyl heptanone, neral, perillene, geraniol, neryl acetate, n-
octyl acetate and geranyl acetate in the essential oils of
cardamom from Reunion. P-menthene and methylallyl benzene were
reported in this oil for the first time.

Unlike steam distilled oils examined by above workers, Bernhard

et all29 studied the oil obtained by cold pressing of ·cardamom
seeds. Besides the above constituents, camphene, a-phellandrene,
trans-sabinene hydrate, DL-camphor, citronella!, 4-terpineol,
cis-and trans-citral, citronellol, 4-terpinyl acetate,
ascaridole, geranyl acetate, bisabolene/(trans-nerolidol) and
farnesol were reported by them.

The essential oil of cardamom is mainly composed of oxygenated

compounds with small amounts of monoterpenes and other minor
constituents (Table 12). Variation in the major constituents
namely 1:8-cineol and a-terpinyl acetate together amounting to
 67

TABLE 12. MAIN COMPONENTS OF CARDAMOM VOLATILE OIL136

Components Area (%}

a-Pinene 1.5
f3-Pinene 0.2
Sabinene 2.8
Hyrcene 1.6
a-Phellandrene 0.2
Limonene 11.6
1,8-Cineole 36.3
Y-Terpinene 0.7
p-Cymene 0.1
Terpinolene 0.5
Linalool 3.0
Linalyl acetate 2.5
Terpinen-4-ol 0.9
a-Terpineol 2.6
a-Terpinyl acetate 31.3
Citronellol 0.3
Nerol 0.5
Geraniol 0.5
Hethyl eugenol 0.2
trans-Nerolidol 2.7

TRACE COMPONENTS IN CARDAMOM VOLATILE OIL

Hydrocarbons Alcohols and Phenols
a-Thujene 3-Methyl butanol
Camphene p-Henth-3-en-1-ol
a-Terpinene Perillyl alcohol
cis-Ocimene p-Cresol
Toluene Carvacerol
p-Dimethylstyrene Thymol
Cyclosativene Carbonyls
a-Copaene 3-Methyl butanal
a-Ylangene 2-Methyl butanal
Y-cadinene Pent anal
D-Cadinene Furfural
Acids 8-Acetoxycarvotanacetone
Acetic Cumlnaldehyde
Propionic Carvone
Butyric Others
2-Nethyl butyric Pinole
3-Methyl butyric Terpinene-4-yl acetate
a-Terpinyl propionate
Dihydro-a-Terpinylacetate
 68

the range of 60-80% was reported among different

varieties138,139.

Expressed oils is found to have lower ranges of a-pinene +

sabinene (2.2-4%) and linalool (2.3-3.8%), normal range of 1:8-
cineol (22.2-33.5%) and distinctly higher range of a-terpinyl
acetate (51.1%-56.4%)128.

1.11.3.2 Phenolic constituents

Schulz and Herrmann45 have isolated and identified nine phenolic

acids accounting for 646 mg/kg in cardamom of which ferulic acid
was the highest in content followed by decreasing order of
vanillic, protocatechuic, p-hydroxybenzoic, p-coumaric, gentisic,
synapic, salicylic and caffeic acids. Hydroxy cinnamoyl quinic
acids and 4- ( 13- D-g 1 ucopyranosyloxy) -benzoic acids were also
reported by Dirks and Herrmann140 while 2-phenyl phenols were
shown to be present in dichloromethane extracts141 of cardamom.
Till date no other report exists on the phenolic constituents of
this spice.

1.11.3.3 Sensory evaluation

Govindarajan et al128 reported that the basic cardamom aroma was

contributed by esters and 1:8-cineol. Lower 1:8-cineol content
with its harsh camphory note together with higher linalyl acetate
content with its sweet-floral odour is responsible for relatively
pleasanter mellow flavour of Malabar variety (Alleppey green),
the largest selling Indian cardamoml28. Minor components modify
and balance the basic character through sweet, spicy, green
floral, woody and sometime lemony and mint notes128. It was
suggested that the expressed oils and cardamom extracts represent
more closely the full rich aroma of original spice as compared to
the usual steam distilled oil128.
 •

69

1.12 NUTMEG AND MACE ( M"Y.RI.STICA

F.RAG.RA~.S)

Nutmeg and mace are two distinct products of the same tree
Hyristica fragrans. While nutmeg is the kernel of the seed, mace
is the dry aril that surrounds the single seed within the fruit.
Host West Indies islands, the Philippines, India, South America
and many pacific islands have climatic conditions that favour
growth of this spice. 40% of the world supply of nutmeg comes
fro m Gr e n ad a \v h ile r e ma i n i n g G0 % c om e s f r o m Ma 1 a y s i a and
Indonesia. India meets most of its demand for nutmeg by imports.

The nutmeg tree Nyristica fragrans is an evergreen tree that

grows to a height of 12 m or more. All parts of the plant are
aromatic. Flowers, which form infloresence are about lcm long,
fragrant, creamy-yellow in colour, waxy and fleshy. Fruits are
orange-yellow in colour and when ripe the yellow-pericarp splits
open to reveal a hard, shiny, grayish brown kernel (nutmeg) and a
lacy red aril (mace). It takes 5-8 years for flowering and 6-9
months from flowering to fruit ripening. Nutmeg in its sh~ll and
the surrounding. aril (mace) is separated from the pericarp of the
harvested fruit and dried separately either artificially or in
the sun. The proportion of dried shelled nutmegs to dried mace
is approximately 20:314.

1.12.1 Products and uses

Primary products of H;vristica fragransl4 which are traded

internationally are (i) Dried nutmeg and mace in the whole/ground
form (ii) oleoresins (iii) essential oils and (iv) nutmeg butter.

In the whole/powdered form nutmeg and mace are used in flavouring

of bakery products, sauces, pickles and in some meat products.
Nutmeg is used mainly for sweet recipes and finds application in
manufacture of processed foods while mace is favoured in savoury
dishes and is preferred for domestic culinary purposes.
 7.0

Nutmeg yields an oleoresin (31-37%) containing substantial amount

of trimyristin when benzene is used as solvent. \vi th c.old
ethanol, however, it yields 18-26% of crude oleoresin. Using
petroleum ether as extracting solvent the yield of mace oleoresin
J.S 27-32% Hot ethanol extracts of mace provide a crude
oleoresin in 22-27% yield. These oleoresins are used in
flavouring foods and in perfumery.

Nutmeg essential oil is used in flavouring processed foods, soft

drinks, pharmaceutical formulations and in perfumery. Hace
essential oil is comparatively very small volume product which is
primarily used for perfumery purposes.

Nutmeg butter is a relatively minor product containing largely

trimyristin having consistency of butter and high proportion of
volatile oil but has limited usage for pharmaceutical
preparations.

1.12.2 Medicinal properties

Both nutmeg and mace are used66 for flatulence, to allay nausea
and vomiting, for convalescents, as an ointment for piles, for
leucorrhea and as a local stimulant to the gastro-intestinal
tract.

1.12.3 Chemistry

Dried nutmeg and mace contains steam volatile oils, phenolics,

fixed oils, proteins, starch, cellulose, pentosans, resin and
mineral elements. Composition of the individual compounds
differs somewhat between the two spices. Both the spices contain
appreciable amounts of fixed oil, their content varying from 25-
40% in case of nutmeg and 20-35% in case of mace; .the composition
of fixed oil is qualitatively similar in both cases but vary
significantly in quantitative terms14,
 71
1.12.3.1 Essential oil

(a) Nutmeg oil

Nutmeg essential oil is a pale yellow thin liquid obtained b

steam distillation in yields of 7-16%14,

Wallach142 as early as 1885 identified limonene and a-pinene i

the essential oil of nutmeg. Nutmeg oil was later on examined b
several workers143-145 in the early 19th century resulting in th
addition of camphene, 13-pinene, borneol, geraniol, linalool, a
terpineol, terpinen-4-ol, p-cymene, eugenol, trans-isoeugenol
myristicin, safrole and myristic acid to this list. With th
advent of chromatographic and spectrometric techniques, th
composition of nutmeg oil was examined in detail. Lee et a114
reported the presence of menthane, menthyl isovalerate, I3
caryophyllene, cumene and cyclaruene aldehyde, while Ikeda et a17
identified 5-3-carene, myrcene, a- and 13-phellandrene
terpinolene, sabinene, a~terpinen and a-thujene in nutmeg oils
Shulgin and co-workers147 reported the identification o
elimicin, cis- and trans-isoelimicin, toluene, trans-methy
isoeugenol and methyl eugenol as constituents of nutmeg oil. Th
studies conducted by later workersl48,149 confirmed the presenc
of all the above mentioned compounds and additionally identifie
J-terpinene' 1: a-cineol' cis-p-ment-2-en-ol' cis-piperi tol, I3
terpineol, cis- and trans-sabinene hydrate, terpinen-4-acetat
and copaene as the constituents of these oils. Schenk an
Lamparsky150 identified 62 constituents 'in nutmeg oil, of whic
a, p-dimethyl styrene, a-cubebene, a-bergamotene, a-humelene, a
farnesene, 13- b i saba lene, 5-cad inene, ge rmacrene D, vani 11 in
fenchyl alcohol, nerol, trans- piperi tel, trans-p-menth-2-en-1
ol, trans-p-menth-2-ene-1, 4 dial, p-cymen-8-ol, citronelly
acetate, neryl acetate, a-terpenyl acetate, 3-methyl-4-decen-l~o
and 3-methyl-4-decenyl acetate were reported for the first tim1
to be present in this oil. Davis and Cook151 recently identifie1
a dehydro diphenyl propanoid derivative of rnyristicin as a ne1
component of nutmeg oil. From the above 1 i tera ture review· i ·

J
 72

appears that the nut~eg essential oil comprises of 64 compound~

while mace essential oil contains 34 compounds. Table U

represents the composition of major components present ir
essential oils of both nutmeg and mace82,

Nutmeg oils are generally classified as East Indian (Indonesia:

or West Indian (mainly from Grenada and St. Vincent) depending or
their geographical origin14, West Indian oils are found to bE
weaker in odour and less spicy compared to East Indian oils whicl
have a higher aromatic ether content and differ in thei1
quantitative composition149,

Oils obtained from oleoresin had a better aroma than stea~

distilled oils152, Higher content of eugenol and phenolic ethe1

derivatives in oleoresins, that are believed to dominate thE
organoleptic profile of the spice is responsible for better aroml
of oils obtained from oleoresin. Use of super critical CO~

extraction provides an oil, free of artifacts153,

(b) Mace oil

Hace oil obtained by steam distillation in yields of 4-17% ha~

similar physicochemical and organoleptic properties to nutrne~

oil14. Shulgin et a]15·1 reported rnyristicin as the rnajo1

constituent of mace oil with safrole and elimicin being presen1
in minor amounts. Detailed study on the composition of mace oi:
by Forrest and Heacock155 and later by Dann et a1156 resulted ir
.the detection of most of the constituents that were also presen1
in nutmeg oil.as summarized in Table 13.

Hace oil .contains 75-94% monoterpene hydrocarbons, 4. 7-17. 6'

oxygenated monoterpenes and sesqui terpenes and 0-5. 9% aromatic
ethers14, Higher amounts of a-pinene and a distinctly highe1
content of aromatic ethers are noted in mace oils compared tc
nutmeg oils14,
 73

TABLE 13. MAJOR CHEMICAL CONSTITUENTS IDENTIFIED IN NUTMEG AND

MACE82

% composition

Compound Rt nutmeg mace

a-Pinene 2.76 14.72 15.24

~-Pinene + Sabinene 3.41 -62.66 45.52
n-Phcllandrene 3.56 3.06 3.17
b-3-Carene 3.78 0.60 0.67
a-Terpinene + p-cymene 3.95 1. 08 3.53
1:8-Cineol + Limonene 4.23 6.18 6.97
u. I. 4.52
~-Phellandrene 4.71 1. 08 2.80
u. I. 4.90 0.28
Y-Terpinene 5.29 0.54 1. 83
Linalool + Terpinolene 5.54 0.48 0.42
~-Terpineol 6.02 0.25 0.32
Borneol (tentative) 6.47 0.05 0.16
u. I. 6.62 0.16
Terpinene-4-ol 7.18 1. 85 4.59
a-Terpineol + Piperitol 7.45 0.36 0.94
U. I. 7.94 0.02 0.27
Geraniol 9.20 0.02 0.22
Safrole + p-cymene-8-ol 9.53 0.53 o. £F
Bornyl acetate 10.18 0.07 - 0. 09
u. I. 10.62 0.08
" 10.95 0.01
" 10.62 0.01
Methyl eugenol i1.26 0.14 0.22
Eugenol + Terpenyl acetate 11.53 0.22 0.15
Geranyl acetate +
a-copaene 12.15 0.29 0.16
Isoeugenol 12.35 0.31 0.45
u. I. 13.13 0.03 0.05
~-Caryophyllene 13.49 0.07 0.07
a-Humulene 14.06 0.02 0.03
u. I. 14.41 0.04 0.08
o-Cadinene 14.53 0.08 0.15
Myristic in 15.08 3.28 5.92
Elemicin 15.76 1. 38 3.14
u.n I. 17.22 0.05 0.11
17.44 0.02 0.18
II
19.20 0.27
n
19.47 0.07
n
21.57 0.03
II
22.10 0.04
Myristic acid 23.12 t 0.01
 .
74

TABLE 13. CONTINUED

% composition
Compound Rt ~utmeg mace

u. I. 23.80 0.03
" 24.42 0.04
" 25.22 0.10
" 25.62 0.08
Trimyristin 26.12 0.06 0.05
u. I. 27.44 0.22
" 28.41
" 29.74 0.02

Rt - Retention time, t - <0.01% U.I. -Unidentified

 7.5

1.12.3.2 Pheno~ic constituents

Schulz and Herrmann45 have reported the isolation an~

identification of seven phenolic acids in nutmeg and eight il
mace amounting to 475 and 1526 mg/kg, respectively. Synapic acic
constituted 73% and 62% of the phenolic acids in nutmeg and mace
respectively. Qualitative variation in the composition oJ
phenolic acids are also noted between the two spices. WhilE
gentisic, caffeic and salicylic acids are absent in nutmeg, macE
is devoid of vanillic and protocatechuic acids. In fact,
gentisic acid is quite predominant in mace. Other phenolic acid~

common to both the spices include ferulic, p-coumaric, p-

hydroxybenzoic and syringic acid. These workerslll have alsc
shown the presence of epicatechin and catechin in nutmeg.
Cyanidine was identified as the major proanthocyanidine of nutme~

responsible for its characteristic astringent taste and colour82.

Several other phenolic constituents such as lignans have bee~

isolated from nutmeg157' 158. Nutmeg contained 2. 5% of total

polyphenols of which 40% comprised of tannins82. Polyphenols
were however, practically absent in mace at full maturity82.

It is important to mention that the scarlet pigment of mace which

greatly influences the commercial value of the spice has been
found to be constituted mainly of lycopene159.

1.12.3.3 Sensory evaluation

Nutmeg flavour has been reported as bitter, warm, spicy, pungent,

heavy, oily and terpeney while the aroma is described as spicy,
warm slightly camphoraceous, sweet and penetrating6. The
essential oil component and phenolic constituents discussed above
are likely to play a major role in imparting these characteristic
flavour and aroma of nutmeg. The strong and coarse flavour note
of nutmeg compared to mace is attributed to higher content of
saturated fatty acid possessing higher melting point that modify
the impression of essential oil on the tongue14.
 76

Correlation between instrumental and sensory techniques have been

carried out to determine the character impact compounds of nutmeg
and mace160. Myristic in, elimicin and safrole were reported to
be the major components responsible for the flavour of the two
spices160.

1.13 ENZYMATIC BROWNING OF

SPICES AND POLYPHENOL
OXIDASE

A prominent feature that occurs during drying of certain spices

is the conversion of fresh green spice to a brown/black finished
product. Oxidation of phenolic compounds mediated either by the
catalytic oxidation by oxidase enzymes such as polyphenol oxidase
or the non-enzymatic autooxidation of labile phenolic compounds
is responsible for this phenomenon161, Studies on these aspects
with respect to fruits and vegetables and model system have been
recent 1 y reviewed by Mach e i x e t a 11 6 2 . However , the
browning/blackening of spices during drying has received little
attention so far. Fundamental step in the chemistry of
browning/blackening is the conversion of o-diphenols to the
corresponding o-quinones163, The o-quinones once formed are very
reactive species which may react with one another, condense with
hydroquinone or condense through two semiquinone radical
intermediates and polymerize to complex brown/black oxidized
products162, In spite of considerable interest with regard to
browning, few studies on polymerization reactions are reported
probably because the intermediate condensation products are
highly labile causing difficulties in their isolation and
characterization.

Polyphenol oxidase ( PPO) ( EC 1. 14.18.1) enzyme widely occurring

in various fruits and vegetables, is a multifunctional copper
containing protein that catalyses both the orthohydroxylation of
monophenols to o-diphenols (cresolase activity) and the
 77

subsequent oxidation of o-diphenols to o-quinones ( catecholase

activity)164, The enzyme has been shown to exist in multiple and
interconvertible forms, many of which differ with regard to their
development, subcellular location and plant source165. The
occurrence, properties, methodology of assay, reaction mechanism
and physiological function of PPO has been reviewed by Mayer
et all65,

Control or prevention of browning in fruit products and a review

of the different methods used has been published recently166.
Inhibition of PPO can be achieved by heat inactivation, by the
exclusion of one or all of the substrates or by the use of
additives. When heat treatment is not desirable, a wide range of
chemical compounds including sulfites, thiol containing
compounds, hal ides, cyanides, acids, diethyldi thiocarbamate and
ascorbic acid have been proposed to inhibit ppol66, Since
cyanides, azides and diethyl dithiocarbamates are toxic and not
permitted for usage in foods, treatment of foods containing PPO
with chemicals such as ascorbic acid, thiol containing compounds
and sulfites is a desirable way to obtain dried products with
good natural appeal. These compounds act either by reducing o-
quinones to o-diphenols or react with o-quinones to yield
colourless compounds. They may also exhibit a direct inhibitory
effect on the enzyme.

1.14 PRESERVATION OF SPICES BY

IONIZING RADIATION

Spices and spice products are prone to insect infestation and

microbial contamination during storage and transportation causing
serious problems in their export. New legal requirements of
spice importing western countries insist that the commodity
should be totally free of microorganisms. Fumigation with
ethylene bromide or ethylene oxide is the only dependable method
being used in the international trade with some degree of
 78

assurance. However, it has several disadvantages in that it doe

not ensure complete control of microorganisms and pests, rna
leave toxic residues and the treated spice can underg
recontamination and cross-contamination on storage. Host of th
importing countries have recently banned this process and ther
is a urgent need for a substitute to decontaminate spices fo
export. Exposure to high energy electromagnetic ionizin
radiations is an unique technique that can be applied fo
microbial decontamination of spices167, Four main sources o
ionizing radiatio11 are employed in food irradiation, two of whic
are the radionucleide or isotope sources ( 60co and 137cs) tha
erni t i -rays \vhile the other t'vo are electrons ( 10 NeV rnaxirnu
energy from a linear accelerator) and X-rays ( 5 MeV maximu
energy from X-ray machine). This technique is a physical proces
like heat or freezing for killing microorganisms and does no
induce radioactivity in foods at doses recommended. 60co is th
major source widely used at present for food irradiation. Th
amount of radiation dose absorbed by a given material is measure
in terms of gray (Gy), It is defined as the amount of radiatio
necessary to deposit one Joule of energy per kg of material. Th
dose range for food applications is measured in terms o
kilograys ( kGy). I\ dose of 10-50 kGy can be used fo
decontaminating spices \vi thou t undesirable effects16 7,
Severa
countries including India have cleared radiation processing fo
decontamination of spices for export purposes.
The maxirnu;
overall average dose allowed for irradiation has been fixed at 11
kGy.

Gamma rays have high penetrating power, there by ensurin1

complete disinfestation of the spice and thus increasing shelf·
life of the product. The method can be applied to prepacked item~
eliminating the risk of recontamination and does not leave an:
toxic residues unlike chemical fumigation. Extensive studies 01
the microbiological, nutritional, toxicological and organoleptic
aspects of irradiated spices have shown that Y-irradiated spice~
treated with ~-rays up to a dose of 10 kGy are wholesome and car
be consumed safely168,
 7.9

Chemiclearance of irradiated food as a supporting evidence t

wholesomeness studies necessitated a detailed chemica
investigation of irradiated foods. Hain components of foo
stuffs are water, carbohydrates, proteins and lipids. Th
radiation chemistry of main food components are elucidated i
literaturel69, For example, gluconic acid, arabinose, 2- and 5
deoxy gluconic acid and 2-deoxyribose were identified as th
radiolytic products of glucose. In addition, the glycosidi
bonds can be broken. Irradiation of starch produces dextrin
maltose and glucose and various other products reducing th
degree of polymerization and viscocity of polysaccharides i
solution. Reaction of lipids or fat portion of foods with
reactive species of water radiolysis plays only a minor role i
most cases. Exposure of triglycerides to ionizing radiatio.
results in formation of cation radical and excited molecule
which undergo deprotonation follo>.,red by dimerization an'
decarboxylation forming alkanes, alkenes, diesters, carbonyls
dicarbonyls and dials. In the case of proteins and amino acids
abstraction of a proton, decarboxylation reactions and th
formation of ammonia and pyruvic acid etc. are the results o
ionizing radiation in protein containing foods. Splitting of S-
bonds, dimerization and formation of other aliphatic amines ar'
important patlnvays in the radiation chemistry of sulphur amin
acids.

Besides the major components mentioned above, chemical changes i:

minor constituents of food such as vitamins, phenolic compounds
essential oils etc. can occur. Thus losses in vitamin conten·
can occur during irradiation. An increase in phenyl alanin1
ammonia lyase (PAL) activity results in an increased conversio1
of phenyl alanine to cinnamic acids and other phenolic compound:
produced from c innamic acid such as p-coumaric acid, caffeic
acid, ferulic acid, glucoside ferulic acid, scopolin anc
scoplectin. For e.g. the cortex of irradiated potatoes was foun<
to contain 50~la more chlorogenic acid and 70% more caffeic aci<
than that of non irradiated potatoes169, No substantial change~
in the· essential oil content were observed in most foc:>d~

J
 80

including spices treated with doses up to 10-15 kGy169.

Preservation of food including spices by ionizing radiation has
been well documented in literature170,171.

1.15 IMPORTANCE OF PHENOLIC

COMPOUNDS

Phenolic constituents present in spices play a major role in the

overall quality on the basis of their astringency, bitterness and
colouring properties as mentioned earlier (Section 1. 7). These
constituents are also important in contributing to the
antioxygenic, antimicrobial and medicinal properties and for
their preservative action3. The antioxidant properties of
phenolic pigment curcumene present in turmeric and of vanillin in
preserving carotenes is reported in li teraturel72' 173. Most of
the antioxygenic and preservative action of clove and thyme is
shown to be due to eugenol and thymol, respectivelyl74.
Herrmann175 reported that the known antioxidant a6tion of clove
oregano, rosemary, sage, thyme, marjoram and savoury is partly
due to labiatic acid, a depside of caffeic acid and p-hydroxy
hydrocaffeic acid. Carvacrol in caraway seeds and eugenol and
eugenol methyl ether in cinnamon and clove possess antimicrobial
and bacteriostatic properties3.

The medicinal properties of spices are likely to be dependent

upon the phenolic constituents present in them. Capsaicinoids
present in red peppers are active ingredients in linaments used
to relieve local muscle or joint pains and are also used the-
rapeutically in the treatment of cutaneous disordersl76. Eugenol,
a volatile phenol, is used as an analgesic to alleviate
toothaches while curcuminoids of turmeric are found to have
anticarcinaogenic and antimutagenic activities176. Tannins or
polyphenols of spices have important pharmacological properties
and are used to control haemorrhage, diarrhoea, inhibit mucous
secretions, as an haemostatic agent, cure disorders of blood
 -81

stream including high BP and in heart diseases177, They also

show antibacterial properties. Phenolic amides and phenolic
amine s including catechol amines are known to be
neurotransmitters, to reduce hypertension, have antibiotic
activities, reduce diabetes and to have antitumor activitiesl78,
Flavanoid glucosides have spasmolytic and diuretic activities eg.
quercetin is kno~"n to be a pmv-erful anticarcinogenl 79. Several
other recent reports on the theurapeutic activities of phenolic
compounds are known in 1 i te ea ture. They include anti virall80,
antiseptic181, antipathogenicl82, antimutagenic183, antitumor184
and antiplatelet aggregation activities185,

The foregoing brief survey of 1 i te ra tu re suggests that a wide

range of volatile and nonvolatile phenolic constituents present
in spices have important biological activities besides their
organoleptic attributes.

1.16 SCOPE OF THE WORK

The chemistry of spice essential oils has been studied by various

researchers and several chemical constituents identified.
However, the knowledge on tlae aroma character impact compound (s)
along with contributory aroma compounds that gives rise to "top
aroma" note of spices is limited. Very few reports are available
on the chemistry of essential oils of green pepper. The nature
of phenolic compounds anu their role in contributing to the
sensory properties of spices has not been thoroughly studied.
Till date no report exists on the nature of phenolic compounds in
green pepper berries. The causative factors involved in the
blackening/broh·ning phenomenon in green pepper and their
chemistry during drying process has not so far been elucidated.

In the present ~ork attempts have been made to identify aroma

character impact and contributory aroma compounds in six
 82
commercially important spices namely pepper, cinnamon, clove
cardamom, nutmeg and mace. Phenolic compounds in these spice:
were characterized and their composition was determined wit:
special emphasis on green pepper berries. Enzymatic blackenin.
of green pepper berries and the involvement of hithertj
unreported phenolic compounds in the drying process have bee.
investigated. Synthesis of these phenolic compounds \-lerj
attempted. Variation in chemical composition with respect t1
volatile flavour compounds, phenolic compounds and pungen·
principles between different cultivars of green pepper were alsj
studied.

The thesis highlights the importance of essential oils an<

phenolic compounds in contributing to aroma, taste, appearancE
and colour and thus to the overall quality of spices. The arom1
impact compound and contributory aroma compounds present in spicE
essential oils appeared to play a major role in contributing tc
the "top aroma" note of these oils. These studies eventuall:
help in identifying better spice cultivars with superior flavouJ
qualities. The presence of two novel phenolic compounds ln fresl
green pepper berries and their unique chemical structure suggest~

the use of green pepper as an effective drug ingredient a~

related compounds are known to have important pharmacologicaJ

properties.