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From Deep Gupta, A, Rajpurohit, D. (2011). Antioxidant and antimicrobial activity of
nutmeg (Myristica fragrans). In V. R. Preedy, R. R. Watson, V. B. Patel (Editors), Nuts
& Seeds in Health and Disease Prevention (1st ed.) (pp 831-839). London, Burlington,
San Diego: Academic Press is an imprint of Elsevier.
ISBN: 9780123756886
Copyright © 2011 Elsevier Inc. All rights reserved
Academic Press
Author's personal copy
CHAPTER 98
Antioxidant and
Antimicrobial Activity
of Nutmeg (Myristica
fragrans)
Ashish Deep Gupta
1
, Deepak Rajpurohit
2
1
Mangalayatan University, Institute of Biomedical Education & Research,
Department of Biotechnology, Uttar Pradesh, India
2
College of Horticulture and Forestry, Department of Biotechnology, Rajasthan, India
CHAPTER OUTLINE
Introduction 831
Botanical Description 832
Historical Cultivation and Usage 832
Present-Day Cultivation and
Usage 833
Application to Health Promotion and
Disease Prevention 833
Antioxidant activity 833
Antimicrobial activity 836
Adverse Effects and Reactions
(Allergies and Toxicity) 837
Summary Points 837
References 838
LIST OF ABBREVIATIONS
BHA, butylated hydroxyanisole
BHT, butylated hydroxytoluene
MMDA, 3-methoxy-4,5-methylene-dioxyamphetamine
TMA, 3,4,5 trimethoxyamphetamine
ROS, reactive oxygen species
INTRODUCTION
Due to potential liver damage and carcinogenic effects, the most widely used synthetic anti-
oxidant compounds, butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA),
have restricted usage (Senevirathne et al., 2006). Therefore, much emphasis is currently being
given to searching for new and natural antioxidants and antimicrobials from dietary plants,
because they can safeguard the human body against the oxidative damage of biological
831
Nuts & Seeds in Health and Disease Prevention. DOI: 10.1016/B978-0-12-375688-6.10098-2
Copyright Ó2011 Elsevier Inc. All rights reserved.
Author's personal copy
macromolecules. Many spices, such as nutmeg (Myristica fragrans), have been reported to have
antioxidant and antimicrobial properties, apart from their traditional use in numerous
medical conditions, and can thus offer a potential solution in the search for new and natural
sources of antioxidants and antimicrobials.
BOTANICAL DESCRIPTION
The nutmeg is a member of the magnoliales order and Myristicaceae family (Figure 98.1).
Nutmeg and mace are two important spices derived from the fruit. Nutmeg is the seed of the
tree; it is dark brown, ovoid, 2e3 cm long, and weighs between 5 and 10 g. Nutmeg seeds
possess ruminate endosperm, and are considered to be the most primitive among the flow-
ering plants. Mace is the dried lacy reddish covering or aril of the seed. A nutmeg tree takes
around 20 years to reach its full potential, but the first harvest can be carried out 7e9 years
after planting. World production of nutmeg is estimated to be 10,000e12,000 tonnes per year,
with annual world demand estimated at 9000 tonnes, whereas production of mace is esti-
mated at 1500e2000 tonnes. The main producers and exporters are Indonesia and Grenada;
India, Malaysia, Papua New Guinea, Sri Lanka, and the Caribbean Islands are other important
producers.
HISTORICAL CULTIVATION AND USAGE
Nutmeg originated in the Banda Islands of Indonesia, and was discovered by the Portuguese in
1512. The importance of the nutmeg seed was propagated by the Dutch. The name nutmeg is
derived from the Latin nux muscatus, meaning “musky nut.” In India, nutmeg is known as
Jaiphal. According to the ethno-medical literature, nutmeg seed oil was used for intestinal
disorders by Indians, in embalming by Egyptians, and to cure plague by Italians. In ancient
times, nutmeg seeds were used in medicines as an aphrodisiac, abortifacient, and anti-flatu-
lent, a narcotic, and as a means to induce menses. The effect of the nutmeg seeds on the central
nervous system was first observed in the early 19th century. Traditional uses of nutmeg seeds
include treatment of hemorrhoids, chronic vomiting, rheumatism, cholera, psychosis,
stomach cramps, nausea, and anxiety. Nutmeg seed oil also has antiseptic, analgesic, and
antirheumatic properties.
FIGURE 98.1
Nutmeg seeds. Nutmeg (Myristica fragrans) is an important spice. Nutmeg is the actual seed, and has been reported to have
strong antioxidant and antimicrobial potential.
832
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PRESENT-DAY CULTIVATION AND USAGE
Both in vitro and in vivo studies have resulted in a wide array of pharmacological actions
attributed to nutmeg seeds, including antioxidant, antifungal, antibacterial (Takikawa et al.,
2002), aphrodisiac (Tajuddin et al., 2003), anti-inflammatory (Olajide et al., 1999), and
hepatoprotective activities. Nutmeg seeds are used to control diarrhea and Crohn’s disease, as
they inhibit prostaglandin activity in the intestine. Nutmeg seeds also improve Alzheimer’s
disease, as they directly affect acetylcholinesterase activity in the brain. Nutmeg seed essential
oil is used in aromatherapy, because the three main constituents of nutmeg (myristicin,
elemicin, and isoelemicin) act as stress relievers.
APPLICATION TO HEALTH PROMOTION AND DISEASE PREVENTION
Volatile essential oil and various extracts of nutmeg seeds have been reported to have many
pharmacological properties, including antioxidant, antimicrobial, insecticidal, anti-amebic,
and anticarcinogenic activity. Nutmeg seed oil is a colorless or pale yellow liquid with the
characteristic odor and taste of nutmeg. The composition of the essential oil of nutmeg seeds
and that of mace differ significantly. The essential oil of nutmeg seeds mainly contains sabi-
nene (15e50%), a-pinene (10e22%), b-pinene (7e18%), myrcene (0.7e3%), 1,8-cineole
(1.5e3.5%), a-phellandrene (0.3e6.2%), myristicin (0.5e13.5%), limonene (2.7e4.1%),
euginol (0.1e1%), safrole (0.1e3.2%), and terpinen-4-ol (0e11%). It has been noted that the
composition of nutmeg seed oil depends upon its source.
Antioxidant activity
In the human body there are various exogenous and endogenous sources for the spontaneous
generation of free radicals and other reactive oxygen species (ROS), such as hydroxyl radicals
($OH), superoxide anions ($O
2
), and hydrogen peroxide (H
2
O
2
), which can affect lipid,
protein, and nucleic acid in various ways and thus play an important role in the initiation and/
or progression of various diseases (Figure 98.2). Under pathological conditions and in
immune-compromised hosts, there is an imbalance between the generation of ROS and their
quenching by the host antioxidant system, which leads to oxidative stress. The exogenous
supply of antioxidants could be very helpful in conquering oxidative stress. Many spices,
including nutmeg, have been reported to possess good antioxidant capacity.
The antioxidant capacity of nutmeg seeds can be measured by various chemical assays, such as
estimation of total phenolic concentration, capacity to scavenge the stable free radical
DPPH (2,2-diphenyl-1-picrylhydrazyl), ferric reducing/antioxidant power assay (FRAP),
inhibition of lipid peroxidation, inhibition of bleaching of b-carotene, etc. The antioxidant
capacity of the essential oil of nutmeg seeds and various extracts has been established by much
research. Jukic et al. (2006) found that the aglycone fraction, enzymatically isolated from
glycosidically bound volatiles of nutmeg, possesses a higher antioxidant capacity compared
with free volatiles from its essential oil. The variation was due to differences in the amounts of
eugenol and isoeugenol. Tomaino et al. (2005) studied the effect of heating on the antioxidant
effectiveness and chemical composition of nutmeg seed essential oil. They reported signifi-
cantly higher free radical scavenger activity with heating, which could be due to the volatili-
zation of the hydrocarbons of the oil at higher temperatures, resulting in the accumulation of
phenolic constituents in the remaining oil.
Antioxidant properties are contributed by the variety of active phytochemicals, including
vitamins, carotenoids, terpenoids, alkaloids, flavonoids, lignans, simple phenols, phenolic
acids, etc. It has been reported that total phenolic content and antioxidant activity have
a significant and positive correlation. In plants, phenolic antioxidants are mainly produced by
secondary metabolism, and their antioxidant property largely depends on their redox properties
and chemical structure (i.e., the number and position of the hydroxyl group). Shan et al. (2005)
reported caffic acid and catechin as the major phenolic acids present in nutmeg seeds.
CHAPTER 98
Pharmacological Importance of Nutmeg
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Compounds like caffic acid, having a catechol structure, are considered to be good antioxidants
because the catechol structure can easily donate phenolic hydrogen or electrons to the accep-
tors, such as reactive oxygen species or lipid peroxyl groups. Chatterjee and colleagues (2007)
reported an antioxidant activity of acetone extract from mace aril, and found that acetone
extract is mainly constituted of lignans. Calliste et al. (2010) reported lignan derivatives as a class
of compounds that contribute to the antioxidant potential of nutmeg seeds. These lignan
derivatives essentially belong to the dibenzyl butane group, with either a guaiacyl or piperonyl
moieties on the aromatic rings. The principal compounds of this category are argenteane (bis-
erythro-5,50-bis [1-(4-hydroxy-3-methoxyphenyl)-4-(3,4-methylenedioxyphenyl)-2,3 dime-
thylbutane]), meso-dihydroguaiaretic acid, and erythro-austrobailignan-6 (Figure 98.3).
Central moieties of these compounds are able to release one or two H atoms to the free radicals,
which can be explained by density functional theorycalculations of the OeH bond dissociation
enthalpies. After absorption into the body, nutmeg seed lignans and their glycosides are
metabolized to produce biologically active compounds containing the catechol structure, which
could account for the high antioxidant potential of the nutmeg seeds (Nakai et al., 2003).
Besides their antioxidant activity, lignans are known to possess diverse pharmacological
potentials, including antitumor, antiviral, and anti-atherosclerotic activities.
Antioxidant activity could be attributed to the occurrence and concentration of various
chemical substances present in the plant. Many compounds that possess good antioxidant
activity have been isolated from nutmeg seeds (Figure 98.4). Compounds such as eugenol
and b-caryophyllene, which contain hydrogen atoms in the benzylic and/or allylic positions,
could be good contributors for antioxidant activity. These compounds have high antioxidant
activity because of the relatively easy abstraction of atomic hydrogen from these functional
groups by peroxy radicals formed under oxidative stress. Another view that favors the
antioxidant role of eugenol in nutmeg could be that it promotes the activities of catalase,
superoxide dismutase, glutamine transferase, glutathione peroxidase, and glucose-6-phos-
phate dehydrogenase enzymes (Kumaravelu et al., 1996).
Reactive oxygen species
Pathogenesis of disease,
Inflammation
phagocytosis
Smoking
Drug toxicity
Heavy metals
Mitochondria
ETS
Endogenous enzymes
NADPH oxidase
Myeloperoxidase
Interaction of ionizing
radiations with
macromolecules
Gastrointestinal
inflammation
Atherosclerosis
Neurodegenerative
Parkinson’s disease
Alzheimer’s disease
Muscular degeneration
Impaired wound healing
Cardiovascular diseases
Ageing,
Cancer
Diabetes
Cataracts
FIGURE 98.2
Synthesis of reactive oxygen species in the human body, and their consequences. In the biological system, there are
many possible ways which can generate ROS. Synthesis of ROS can lead to various life threatening diseases.
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O
H3C
HO
CH3
CH3
OH
O
CH3
Meso-di hydroguaiaretic acid
O
H3C
HO
CH3
CH3
O
O
Erythro-austrobailignam-6
Argenteane
O
O
H3C
H3C
O OH
CH3
OH O
CH3
CH3
CH3
O
O
FIGURE 98.3
Important Lignans of the nutmeg seed. Meso-di hydroguaiaretic acid, erythro-austrobailignan-6 and argenteane
(bis-erythro-5,50-bis [1-(4-hydroxy-3-methoxyphenyl)-4-(3,4-methylene dioxyphenyl) 2,3 dimethylbutane]) are major lignans in
nutmeg. Lignans are important antioxidants, as they can release H atoms to the free radicals.
FIGURE 98.4
Important constituents of nutmeg seed oil. b-Caryophyllene [trans-(1R,9S)-8-Methylene-4,11,11-trimethyl bicycle [7.2.0]
undec-4-ene], safrole [3,4-methylenedioxyphenyl-2-propene], eugenol [2-Methoxy-4-(2-propenyl) phenol], and isoeugenol
[2-Methoxy-4-(1-propenyl) phenol] exert antioxidant activity via various mechanisms. b-Phellandrene [3-methylene-6-
(1-methyl ethyl) cyclohexene] is the main aphrodisiac compound of nutmeg.
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Antimicrobial activity
Various extracts and the essential oil of nutmeg seeds have presented strong antimicrobial
activity against gram-positive and gram-negative bacteria, as well as a variety of fungi. Takikawa
et al. (2002) reported antimicrobial activity of ethanolic extract of nutmeg seeds against entero-
hemorrhagic E. coli O157, which was found to be highly sensitive to b-pinene. Narasimhan
and Dhake (2006) reported potent antibacterial activity of chloroform extract of nutmeg seeds
against both gram-positive and gram-negative bacteria. They found trimyristin and myristic
acid to be the chief antibacterial principles isolated from nutmeg seeds. Cho et al. (2007)
isolated three lignans (erythro-austrobailignan-6, meso-dihydroguaiaretic acid, and nectan-
drin-B) from the methanolic extract of nutmeg seeds, which were reported to have antifungal
activity. These three lignans were found to suppress the development of rice blast and wheat leaf
rust. Some important antimicrobial compounds reported in nutmeg seeds are a-pinene,
b-pinene, p-cymene, carvacrol, and b-caryophyllene, (Dorman & Deans, 2004)(Figure 98.5).
Many plant phenolics have been reported to possess antimicrobial activity. b-Caryophyllene
has been reported to have anti-inflammatory and antifungal activities (Sabulal et al., 2006).
a-Pinene and b-pinene (pinene-type monoterpene hydrocarbons) have been reported to have
antimicrobial activity (Dorman & Deans, 2000), and are supposed to be involved in membrane
disruption by the lipophilic compounds. Another important component for antimicrobial
activity could be carvacrol. The mode of action of carvacrol on bacteria is similar to that of other
phenolic compounds, and occurs via membrane damage, resulting in an increase in membrane
permeability to protons and potassium ions, depletion of the intracellular ATP pool, and
disruption of the proton-motive force. p-Cymene could also be an important component,
because it is a precursor of carvacrol. It has been reported that p-cymene shows weak anti-
bacterial activity when used alone, but works synergistically with carvacrol in expanding the
membrane, which in turn causes destabilization of the membrane (Ultee et al., 2002). It has
been suggested that antimicrobial activity could be attributed to both major and minor
components; it is possible that the antimicrobial activity of major components is regulated by
FIGURE 98.5
Important antimicrobial compounds of nutmeg seed. a-Pinene, b-pinene, p-cymene [1-methyl-4-(1-methylethyl)-
benzene], and carvacrol [2-methyl-5-(1-methylethyl)-phenol] are the chief antimicrobial compounds of nutmeg. The main
mechanisms for antimicrobial activity are membrane disruption, or depletion of the intracellular ATP pool.
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some other minor components, as well as these minor components being able to interact with
other components to exert antimicrobial activity (Bounatirou et al., 2007).
ADVERSE EFFECTS AND REACTIONS (ALLERGIES AND TOXICITY)
Consumption of nutmeg seeds in high dosage has been reported to lead to facial flushing,
tachycardia, hypertension, dry mouth, blurred vision, psychoactive hallucinations, feelings of
euphoria and unreality, and delirium. Recently, several cases of nutmeg seed ingestion have
been reported in adolescents in particular, all of whom were attempting to achieve a euphoric
state at low cost (Demetriades et al., 2005). Symptoms usually begin about 3e6 hours after
ingestion, and resolve by 24e36 hours. The medical literature does not cite any fatalities solely
related to nutmeg intoxication. The possible cause for the psychoactivity of nutmeg seeds
could be metabolic conversion of elemicin and myristicin into amphetamine-like compounds
(Figure 98.6). Elemicin is observed to metabolize to 3,4,5 trimethoxyamphetamine (TMA),
and myristicin to 3-methoxy-4,5-methylene-dioxy amphetamine (MMDA), which are
amphetamine derivatives (Stein et al., 2001). Moreover, myristicin is a weak inhibitor of
monamine oxidase, which could be responsible for some cardiovascular symptoms.
SUMMARY POINTS
lDue to potential liver damage and carcinogenic effects, most widely used synthetic
antioxidant compounds such as butylated hydroxytoluene and butylated hydroxyanisole
have restricted usage.
lNutmeg is the dried kernel of a broadly ovoid seed. Nutmeg seeds have been reported to
have antioxidant and antimicrobial properties, besides their use as an important folk
medicine.
FIGURE 98.6
Metabolism of elemicin and myristicin in 3,3,5-trimethoxyamphetamin (TMA) and 3-methoxy-4,5-methylendioxy
amphetamine (MMDA). 3,3,5-Trimethoxyamphetamin (TMA) and 3-methoxy-4,5-methylendioxy amphetamine (MMDA) are
major compounds that cause hallucinogenic effects.
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lNutmeg seeds possess a high antioxidant potential, which is attributed to caffic acid,
catechin, eugenol, b-caryophyllene, argenteane, meso-dihydroguaiaretic acid, and erythro-
austrobailignan-6.
lNutmeg seeds show strong antimicrobial activity against gram-positive and gram-negative
bacteria, as well as against various pathogenic fungi. Antimicrobial activity is contributed
by b-caryophyllene, a-pinene, b-pinene, p-cymene, and carvacrol.
lDue to its high antioxidant and antimicrobial activities, nutmeg could be considered as
a significant natural source of antioxidants and antimicrobials. Nutmeg, being a natural
product, can offer more safety to people and the environment, and is considered to be less
of a risk for resistance development by pathogenic microorganisms.
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