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Pharmacological Activities of Basil Oil

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10

Pharmacological Activities of Basil Oil:

A Review
NASSER A. AWADAH ALI1*AND WILLIAM N. SETZERB2

ABSTRACT
Basil is a well-known aromatic plant used in the traditional medicine
in many Asian, African, and South American countries for treatment of
diseases. The therapeutic activity of basil is partly due to the presence of
essential oils which possess a wide spectrum of pharmacological effects.
In the present review, efforts have been made to sum up different aspects
of scientific studies on the basil essential oil such as ethnopharmacology,
phytochemistry and mainly the pharmacological activities. In vitro and
in vivo studies, for various pharmacological activities of the basil oil
e.g., antioxidant, antimicrobial, anticarcinogenic, anti-inflammatory,
immunomodulatory, muscle relexant, antiulcerogenic, mosquito
repellent, protozoacidal, and nematicidal etc., have been demonstrated.

Key words: Ocimum, Antimicrobial, Antioxidant, Cytotoxic, Essential

oils

INTRODUCTION

Ocimum is a genus of annual and perennial herbs and shrubs found in

the tropical and subtropical regions of Asia, Africa and South America

1
Pharmacognosy Department, Faculty of Pharmacy, Sana'a University, P. O. Box
13150, Sana'a, Yemen
2
Department of Chemistry, University of Alabama in Huntsville, Huntsville,
AL35899, USA
* Corresponding author: E-mail: [email protected]
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286 RPMP Vol. 37: Essential Oils–II

(Darrah, 1988). It belongs to the Lamiaceae, one of the largest and

most distinctive families of flowering plants, with about 220 genera
and almost 4000 species worldwide. The taxonomy of Ocimum is complex
due to interspecific hybridization and polyploidy of the species within
the genus. Pushpangadan and Bradu (1995) recognized more than 150
species; however, (Paton et al., 1999) proposed that Ocimum had only
65 species and other attributions should be considered as synonyms.

The genus Ocimum comprises important medicinal plants, which

are currently the subject of phytochemical attention because of their
biological and chemical diversity and essential oil production, a few of
which are O. basilicum,O. americanum, O. gratissimum, O. canum, O.
minimum, O. urticifolia, O. forskolei, O. tenuiflorum, O. suave, and O.
spicatum. Nowadays, the plant is extensively cultivated for its essential
oil in several countries. Information regarding the ethnopharmacology,
phytochemistry, pharmacological activities involving in vitro and in
vivo experiments, and a few human studies available in the recent
scientific literature is presented.

HISTORY AND ETHNOPHARMACOLOGY OF BASIL

Basil has long been used as an embalming and preserving herb, found
in mummies of ancient Egypt. Basil was used also a symbol of mourning
in Greece where it was known as basilikon photon, meaning magnificent,
royal, or kingly herb. Ancient records from 907 A D indicate sweet basil
in the Hunan region of China. Many beliefs and rituals are accompanied
with basil. In Italy it is a symbol of love, in France it is named as an
herb of royalty. Jewish people used it to get strength during fasting
while an African legend claims that basil protects against scorpions.
However, a European group has considered it to be a symbol of Satan
(Marwat et al., 2011).

The plant has been used in many parts of the world to treat a wide
variety of disorders. Basil is a popular herb in US and Mediterranean
diets (Lee & Scagel, 2009). O. basilicum, sweet basil is used in Yemeni
traditional medicine to treat various ailments: abdominal cramps,
gastroenteritis, dysentery, and diarrhea. In northern Oman and Saudi
Arabia, juice of leaves or crushed leaves is used in the treatment of
wounds, acne, and vitiligo. It is used also as a deodorant, is considered
to be an aphrodisiac, worn by men when visiting their women
(Ghazanfar, 1994; Schopen, 1983).

In the coastal areas of Nigeria, O. gratissimum is used in the

treatment of epilepsy, high fever and diarrhoea (Effraim et al., 2003).
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Pharmacological Activities of Basil Oil: A Review 287

O. gratissimum is used by the Ibos of Southeastern Nigeria in umbilical

cord management, to keep wound surfaces sterile, as well as the
treatment of fungal infections, fever, cold and catarrh (Ijeh et al., 2005).
The infusion of O. gratissimum leaves is used as pulmonary
antisepticum, antitussivum and antispasmodicum (Ngassoum et al.,
2003). Brazilian tropical forest inhabitants use a decoction of O.
gratissimum roots as a sedative for children (Cristiana et al., 2006). In
India, the whole plant has been used for the treatment of sunstroke,
headache, influenza, as a diaphoretic, antipyretic and for its anti-
inflammatory activity (Ueda-Nakamura et al., 2006).

Juice of the leaves of Tulsi plant, O. sanctum was used as demulcent,

stimulant, expectorant. Tulsi was also used in the cure of upper
respiratory tract infections, bronchitis, skin infections and earache
(Harsa et al., 2003).

Traditionally, extracts of O. kilimandscharicum have been used to

mitigate many disorders in East Africa including coughs, colds, measles,
abdominal pains, and diarrhea. It has also been used as an insect
repellent, particularly against mosquitoes and food storage pests (Bekele
et al., 1995). The essential oils obtained from this plant have been used
in northeastern Tanzania for centuries as repellents against nuisance
biting insects and malaria vectors (Kweka et al., 2008).

O. canum is used for treating various types of diseases, lowering

blood glucose, and for treatment of colds, fever, parasitic infestations
on the body, and inflammation of joints and headaches (Ngassoum et
al., 2003). Traditionally, Generally Ocimum species have been
extensively utilized in food and perfumery industries (Telci et al., 2006).

PHYTOCHEMISTRY

Ocimum is noted for its pungency and flavor apart from its aroma. The
aroma in this genus is due to an essential oil, the contents of which
range from 0.3 to 3.6% (dry wt. basis). The major essential oil
components include phenyl-propanoids (methyl chavicol, methyl
cinnamate, eugenol, and methyl eugenol) and terpenes (linalool,
geraniol, geranial, camphorand neral). The minor components in this
genus, most of which are sesquiterpenes, are found to vary amongst
species. Basil has been classified according to different geographical
origins. They are the European chemotype, from Italy, France, Bulgaria,
Egypt, and South Africa, having linalool and methyl chavicol as main
components; tropical chemotype, from India, Pakistan and Guatemala,
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288 RPMP Vol. 37: Essential Oils–II

being rich in methyl cinnamate; Reunion chemotype, from Thailand,

Madagascar and Vietnam, being characterized by high concentration
of methyl chavicol. There is also a eugenol-rich chemotype from North
Africa and Russia. Other chemotypes were reported in recent studies
such as ß-caryophyllene in O. sanctum and O. micranthum (syn. O.
campechianum); citral in O. citriodorium and O. canum; ethyl cinnamate
in O. gratissimum; 1,8-cineole in O. micranthum (syn. O. campechianum;
thymol in O. gratissimum; p-cymene in O. gratissimum; geranyl acetate
in O. minimum; and camphor in O. canum (Telci et al., 2006; Grayer et
al., 1996).

PHARMACOLOGICAL ACTIVITIES OF BASIL OIL

Antioxidant Activity

Some compounds found in Ocimum plant oil have been found to possess
strong antioxidant activity. Essential oils containing higher levels of
linalool or eugenol should provide a stronger antioxidant activity as
was found by Julisni and Simon, 2002. In other study O. basilicum oil
with linalool, isoanethole and eugenol as major components has shown
appreciable antioxidant activities, comparable to that of á-tocopherol
(Wei et al., 2010). Similarly, the essential oils of basil have demonstrated
radical-scavenging and antioxidant properties in the 2,2-diphenyl-1-
picrylhydrazyl (DPPH) radical scavenging assay at room temperature
(Tomaino et al., 2005). Hussain (2009) reported that O. basilicum
essential oil (OBEO) collected from hilly and plains regions showed
highest radical scavenging activity (IC50 8.31-10.1 µg/ml) followed by
O. gratissimum (OGEO) (IC50 6.7-8.2 µg/ml) and O. sanctum (OSEO)
(IC50 11.1-13.5 µg/ml). In the same study, within different species of
Ocimum, OBEO (78.0-84.2%) showed better inhibition of peroxidation
of linoleic acid system than OGEO (77.2-81.5%) and OSEO (70.1-74.0%).
The major component of OBEO, linalool, also exhibited good antioxidant
activity (75.2%). Similarly, the antioxidant activities of OBEO var.
thyrsiflora and O. basilicum leaves were tested by DPPH radical
scavenging. OBEO (IC50 = 26.53 ± 0.94 µg/ml) exhibited a higher
scavenging ability for DPPH radicals than OBEO var. thyrsiflora (IC50
= 98.33 ± 2.08 µg/ml) (Pripdeevech et al., 2010).

Al-Maskri et al. (2011) studied the chemical composition and the

ABTS-scavenging ability of Omani OBEO. Total antioxidants content
quantified in Omani OBEO using a commercial kit were found to be
50.32 ± 1.8 mM. Omani OBEO was found to have higher antioxidant
content than previously reported values of many cultivars in literature.
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Pharmacological Activities of Basil Oil: A Review 289

Politeo et al. (2007) investigated antioxidant activity measured by DPPH

radical scavenging and ferric reducing/antioxidant power assay (FRAP)
of OBEO and free volatile aglycones. Eugenol, chavicol, linalool and á-
terpineol were the common compounds. DPPH method shows that free
volatile aglycones possess good antioxidant properties comparable with
that of the essential oil and well-known antioxidant butylated
hydroxytoluene (BHT), but less than pure eugenol. The results obtained
by FRAP method show that these compounds are somewhat less effective
antioxidants than the essential oil or BHT.

Suanarunsawat et al. (2010) have investigated the anti-

hyperlipidemic and antioxidant activities of essential oils extracted from
O. sanctum L. leaves in rats fed with high cholesterol diet. Eugenol and
methyl eugenol were the most important components in the essential
oils. The oils were able to decrease the high levels of thiobarbituric acid
reactive substances (TBARS) either in cardiac or liver tissues, protecting
them against stress induced oxidation.

In vitro antioxidant activity of the O. canum oil showed dose-

dependent free-radical scavenging activity against DPPH (IC50 523.55
± 0.001 µg/ml) and hydroxyl radicals (491.12 ± 0.002 µg/ml), metal
chelating (781.38 ± 0.001 µg/ml) and prevention of deoxyribose
degradation (168.50 ± 0.003 µg/ml) activities were also calculated.
Antioxidant activity of oil was less observed when compared with
positive controls such as ascorbic acid and BHT (12.49 ± 0.002 and
30.14 ± 0.005 µg/ml) respectively (Selvi et al., 2012).

According to Bunrathep et al. (2007), the free radical scavenging

capacity of O. gratissimum oil was higher than that of either O. basilicum.
O. canum, O. sanctum or tocopherols. In their experiment, O. gratissimum
oil reduced the radical generator 2,2-diphenyl-1-picrylhydrazyl (DPPH)
by 50% (IC50 30.20 µg/ml) compared to tocopherols (IC50 62.77 µg/ml). A
study conducted by Leelapornpisid et al. (2008) revealed that holy basil
oil (O. sanctum) exhibits the highest antioxidant activity with IC50 of
0.6294 mg/ml compared to other oils tested in DPPH-method.

The phenolics and essential oil from O. kilimandscharicum leaves

exhibited antioxidant activity estimated by TBARS assay in vitro and
enzymatic antioxidant estimation in vivo (Singh et al., 2011).

Antimicrobial Activity

Essential oil present in most of the Ocimum species is responsible for

its antifungal, antibacterial and antiviral properties. The essential oils
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290 RPMP Vol. 37: Essential Oils–II

of various Ocimum species have been shown, in vitro, to have

antibacterial activity against Staphylococcus aureus, Salmonella
enteritidis, Escherichia coli, Proteus vulgaris, Bacillus subtilis,
Salmonella typhi, Shigella sonnei, S. bodyii, Pseudomonas aeruginosa
and Salmonella paratyphi (Sinha & Gulati, 1990).

The antibacterial properties of O. basilicum essential oil have been

studied using the agar disk diffusion assay with Gram-negative bacteria
Escherichia coli, Pseudomonas aeruginosa, and Gram-positive Bacillus
cereus, Staphylococcus aureus. Both minimum inhibitory concentrations
(MIC) and minimum bactericidal concentrations (MBC) were
determined. The results of agar disk diffusion tests showed at
concentration of 30 µL the inhibition zones as follow: S. aureus 29.20-
30.56 mm, B. cereus 10.66-16.11 mm, E. coli 17.48-23.58 mm and for P.
aeruginosa the maximum inhibition zones were seen. The results of
this study showed the presence of bacteriostatic effects of basil essential
oil on all the test bacteria. The MICs for gram-positive bacteria were
determined to be: B. cereus ranging 36-18 µg/ml, S. aureus 18 µg/ml,
and for Gram-negative bacteria of E. coli and P. aeruginosa were 18-9
µg/mL (Moghaddam et al., 2011). In another study, (Sinha & Gulati,
1990) have reported the effectiveness of the O. basilicum oil on E. coli,
Salmonella typhi, S. paratyphi, Shigella boydi, Proteus vulgaris, and
S. aureus. An in vitro study revealed that Propionibacterium acnes was
more susceptible to O. basilicum oil than other Ocimum species
(Pansanga et al., 2010).

Kristinsson et al. (2005) reported that the treatment of experimental

acute otitis media with oil of sweet basil or essential oil components
cured or healed 56%-81% of rats infected with Haemophilus influenzae
and 6%-75% of rats infected with pneumococci, compared with 5.6%-
6% of rats in the placebo group. Essential oils or their components placed
in the ear canal can provide effective treatment of acute otitis media.

Essential oils of O. basilicum, O. sanctum and O. gratissimum were

tested against 13 bacterial strains and 10 fungal strains (Hussain, 2009).
O. sanctum essential oils were found to be comparatively more active
against the bacterial strains tested, particularly against Gram-positive
bacteria as well as against fungi. Results obtained from disc diffusion
method, followed by determination of minimum inhibitory
concentrations, indicated that generally S. aureus, Bacillus pumilis, B.
subtilis and B. cereus were the most sensitive microorganism tested
with the largest zones of inhibition, 18.7-27.1, 19.0- 25.1, 19,7-27.2,
19.0-26.4 mm, and lower MIC values, 787.2-1097.0, 840.0-1100.0, 776.0-
1100.6, 833.3-1108.0 µg/ml respectively. Among fungal strains, Mucor
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Pharmacological Activities of Basil Oil: A Review 291

mucedo was the most resistant strain while Candida krusei was the
most sensitive strain tested. Linalool, the major component of the
essential oil of O. basilicum, showed stronger antibacterial activity than
antifungal activity with MIC values range from 304.5 to 909.2 µg/ml
and 1009.5-1635.4 µg/ml, respectively (Hussain, 2009).

The antimicrobial activity of six essential oils from four Tanzanian

Ocimum species, O. basilicum, O. kilimandscharicum, O. lamiifolium,
and O. sauve, against four Gram-positive bacteria S. aureus, S.
epidermidis, Streptococcus mutans and S. viridans, four Gram-negative
bacteria P. aeruginosa, Enterobacter cloacae, Klebsiella pneumoniae and
E. coli and three species of the yeast Candida, C. albicans, C. tropicalis,
and C. glabrata, was investigated. The oil of O. suave (sample B) showed
the strongest antibacterial activity on all tested bacteria with MIC
ranging from 0.05 to 1.45 mg/ml; this could be attributed to its major
component, methyl eugenol (82.7%). O. suave (sample A), O.
kilimandscharicum and O. lamiifolium were moderately active, while
O. basilicum oil was weakly active (Runyoro et al., 2010). In another
study, antibacterial activity was done against a number of common
pathogens: Enterococcus faecalis, E. coli, Listeria monocytogenes, L.
ivanovii, Proteus vulgaris, S. aureus, and S. epidermis. E. coli was
inhibited by O. basilicum while O. americanum and O.citriodorum
essential oils were the most effective against E. faecalis, E. faecium, P.
vulgaris, S. aureus and S. epidermis. Methyl chavicol, linalool, methyl
eugenol and methyl cinnamate, isolated from aerial parts of O.
basilicum, O. kilimandscharicum, and O. gratissimum, exerted
antimicrobial activity evaluated by broth dilution method (Anand et
al., 2011). The essential oils of the aerial parts of O. basilicum var.
pilosum showed significant antifungal activity against the
phytopathogenic fungi Fulvia fulva, Glomerella cingulata, Alternaria
alternate and Fusarium solani with IC50 of 15.58, 363.92, 376.70, and
14.27 µg/ml, respectively. Two components in this herb, linalool (29.68%)
and methyl (Z)-cinnamate (21.49%) accounted for the highest percentage
(Zhang. et al., 2009).

The O. basilicum oil was tested against pathogenic fungi viz.

Aspergillus niger, A. fumigatus, Penicillium italicum and Rhizopus
stolonifer using a disc diffusion method, and by determination of
minimum inhibitory concentration. Surprisingly high antifungal values
were found highlighting the potential of Omani basil as a preservative
in the food and medical industries (Al-Maskri et al., 2011).

Studies have shown O. basilicum act as a strong antiviral agent

against DNA viruses [herpes simplex viruses (HSV), adenoviruses
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292 RPMP Vol. 37: Essential Oils–II

(ADV) and hepatitis B virus (HBV)] and RNA viruses [coxsackievirus

B1 (CVB1) and enterovirus71 (EV71)] O.tenuiflorum also has been
reported to be having antiviral activity against bovine herpesvirus 1
(BHV-1) (Chiang et al., 2005).

The essential oil obtained from Kenyan O. gratissimum was

characterized by a high percentage of eugenol (68.8) and methyl eugenol
(13.21%). The antimicrobial activities of the essential oils were evaluated
against both Gram positive (S. aureus, Bacillus spp.) and Gram negative
(Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, Klebisiella
pneumoniae, Proteus mirabilis) bacteria and Candida albicans. The
essential oil showed significant activity on all the Gram negative
bacteria including those which were resistant (P. aeruginosa, S. typhi,
and P. mirabilis) to reference antibiotic (chloroamphenicol) (Matasyoh
et al., 2007). In another study, O. gratissimum essential oil was also
shown to inhibit extracellular protease and the expression of O-
lipopolysaccharide rhamnose in virulent and multidrug-resistant strains
of 22 Shigellae (Iwalokun et al., 2003). Thus, the oil may find a use as a
therapeutic measure against shigellosis.

Study was carried out to investigate the effect of honey as well as

those of surfactants on the antibacterial activity of the essential oil of
O. gratissimum. The antibacterial activity of dispersions of Ocimum
oil (2%) in methanol, honey, a macrogol blend, nonionic and ionic
emulsifiers were assessed by cup–plate method using type bacteria and
wound isolates. Honey enhanced the antibacterial activity of Ocimum
oil to a greater extent than the macrogol blend. The activity of Ocimum
oil emulsion in cetrimide (cationic) was lower than obtained for cetrimide
solution. Emulsion of the oil in sodiumlauryl sulphate (anionic) exhibited
a slightly high eractivity than the solution of the surfactant alone.
Although Tween® 20 (nonionic) and aqueous methanol had no activity,
the emulsion of the oil in Tween® 20 showed lesser activity than the oil
solution in methanol. Honey’s inherent antibacterial activity, surfactant
charge interaction and the effect of emulsification were adduced to the
observed differences in antibacterial activity of the Ocimum oil
formulations. The findings indicated that honey was a suitable base for
Ocimum oil especially in the treatment of infected wounds (Orafidiya
et al., 2006).

The antibacterial activity of different extracts from the leaves of O.

gratissimum was tested against S. aureus, E. coli, Salmonella typhi
and Salmonella typhimurium, pathogenic bacteria that cause diarrhoea.
Extracts evaluated included cold water extract, hot water extract and
steam distillation extract. Only the steam distillation extract had
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Pharmacological Activities of Basil Oil: A Review 293

inhibitory effects on the selected bacteria and the minimum inhibitory

concentration ranged from 0.1% for S. aureus to 0.01% for E. coli and S.
typhimurium, and 0.001% for S. typhi (Adebolu & Salau, 2005).

Mbata and Saikia (2007) showed that O. gratissimum oils have

properties that can inhibit the growth of psychrophils and heat resistant
organisms and suggested that the plant and its derivatives can be used
for the primary purpose of flavouring foods and for antimicrobial
activities. Antibacterial activity of the essential oil of hydrodistilled
leaves of O. gratissimum on oral bacterial isolates was investigated
using the agar diffusion method. The oil inhibited the growth of
Streptococcus mutans, Staphylococcus aureus, Staphylococcus
epidermidis Lactobacilluscasei, Streptococcus sanguis, Streptococcus
salivarius, and Streptococcus pyogenes while no activity was recorded
against Streptococcus viridans or Actinomyces israeli (Saliu et al., 2011).

Essential oils extracted by hydrodistillation from local plants in

Benin, western Africa were evaluated in vitro and in vivo for their
efficacy against Fusarium verticillioides infection and fumonisin
contamination. O. gratissimum was found to be the most effective in
vitro, completely inhibiting the growth of F. verticillioides at lower
concentrations over 21 days of incubation. These oils reduced the
incidence of F. verticillioides in corn and totally inhibited fungal growth
at concentrations of 8, 6.4, and 4.8 µl/g, respectively, over 21days. At
the concentration of 4.8 µl/g, these oils did not affect significantly
fumonisin production. However, a marked reduction of fumonisin level
was observed in corn stored in closed conditions. The oils adversely
affected kernel germination at 4.8 µl/g and therefore cannot be
recommended for controlling F. verticillioides on stored corn used as
seeds, when used at this concentration (Fandohan et al., 2004).

The antifungal activity determined by the dilution method and

expressed as a Number of Decimal Reduction of the colony forming
units per ml (NDR cfu) showed that the essential oils extracted from O.
gratissimum was significantly (P < 0.05) more active against Penicillium
expansum than those extracted from C. citratus and T. vulgaris.
Fractions enriched with oxygenated terpenes were significantly (P <
0.05) more active than their respective essential oils, whereas most of
the fractions enriched with terpene hydrocarbons, were significantly
(P < 0.05) less active. Penicillium expansum is a mould that causes the
rotting of several fruits and vegetables, especially apples onto which it
also synthesizes some dangerous mycotoxins (Nguefack et al., 2012). In
another study, an investigation of antifungal activity of the essential
oil obtained by steam-distillation (1.1% w/w) of the aerial parts of O.
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294 RPMP Vol. 37: Essential Oils–II

gratissimum and of an ethanolic extract from the steam-distillation

residue was carried out using the agar diffusion method. The results
revealed that the essential oil inhibited the growth of all fungi tested,
including the phytopathogens, Botryosphaeria rhodina, Rhizoctonia sp.
and two strains of Alternaria sp., while the extract from the residue
was inactive. The antifungal activity of eugenol was evaluated against
a species of Alternaria isolated from tomato and Penicillium
chrysogenum. The minimal inhibitory concentrations (MIC) of eugenol
were 0.16 and 0.31 mg/disc for Alternaria sp. and P.chrysogenum,
respectively (Terezinha et al., 2006).

The hexane extract of O. gratissimum leaves and eugenol were

investigated for in vitro antifungal activity, using the agar dilution
technique against dermatophytes. The extracts (hexane, chloroform
fractions, the essential oil and eugenol) produced antifungal activities
against Microsporum canis, M. gypseum, Trichophyton rubrum and T.
mentagrophytes. The hexane fraction and eugenol were the most active.
The hexane fraction inhibited the growth of 100% of dermatophytes at
a concentration of 125 µg/ml, while eugenol inhibited the growth of
80% of dermatophytes at this same concentration. These results show
that extracts of O. gratissimum are active in vitro against human
pathogenic dermatophytes (Silva et al., 2005).

In another study, antifungal activities were carried out by the agar

dilution method using five chemotypes of O. gratissimum. Out of these
five chemotypes, ethyl cinnamate showed better activity and was active
against dermatophytes and Scopulariopsis breicaulis, causing skin
mycosis and onychomycosis; against Cryptococcus neoformans,
implicated in HIV disease, and against Malassezia found in the dog’s
otitis externa. Due to these polyvalent performances and the sweet
fragrance of this natural product, O. gratissimum essential oil containing
a high level of ethyl cinnamate seems especially suitable fordermatology
and cosmetology (Dubey et al., 1997).

The GC-MS analysis of O. sanctum essential oil showed a high content

of methyl chavicol (44.63%) and linalool (21.84%). Study revealed that
O. sanctum essential oil has significant antifungal activity against
Candida and hence it can be applied in treatment of fungal infections.
The antifungal activity of O. sanctum essential oil was enhanced by its
synergy with established azole antimycotics-fluconazole and
ketoconazole against seventy four fluconazole-sensitive and sixteen
fluconazole resist Candida isolates. Thus, O. sanctum essential oils,
extracts, or in combination are promising antifungal agents for
treatments of candidiasis (Khan et al., 2010).
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Pharmacological Activities of Basil Oil: A Review 295

O. sanctum essential oil and its major component eugenol were found
to be active against the fungi causing bio-deterioration of food stuffs
during storage. O. sanctum and eugenol were found effective in
inhibiting the growth of Aspergillus flavus NKDHV8; and, their
minimum inhibitory concentrations (MICs) were recorded as 0.3 and
0.2 µg/ml respectively. The LD50 value of O. sanctum essential oil on
mice was found to be 4571.43 µg/kg (Kumar et al., 2010).

Essential oil from aerial parts of O. kilimandscharicum showed

antimicrobial activity against Gram-positive bacteria (S. aureus,
Enterococcus faecalis), Gram-negative bacteria (E. coli, P. aeruginosa)
and also against yeast C. albicans (Kumar et al., 2011).

Cytotoxic activity

Taie et al. (2010) studied the cytotoxic effect for different concentrations
of O. basilicum ethanolic extract and O. basilicum essential oil in Ehrlich
ascites carcinoma cells by the modified cytotoxan trypan blue-exclusion
technique. Ocimum oil recorded the highest inhibition in the viability
of the cells with IC50 of 0.0616 µg/ml (ppm). These results incorporate
the findings by (Manosroi et al., 2006) who reported that sweet basil
leaf oil gave the IC 50 value of 0.0362 µg/ml in P388 cell line. The
dominant constituent in basil oil was linalool (88%). Kathirvel and Ravi
(2012) conducted the studies to identify the chemical composition and
in vitro anticancer activity of the essential oil from O. basilicum Linn.
The major constituents were found to be methyl cinnamate (70.1%),
linalool (17.5%), b-elemene (2.6%) and camphor (1.52%). The results
revealed that this plant may belong to the methyl cinnamate and linalool
chemotype. A methylthiazole tetrazolium assay was used for in vitro
cytotoxicity screening against the human cervical cancer cell line (HeLa),
human laryngeal epithelial carcinoma cell line (HEp-2) and NIH 3T3
mouse embryonic fibroblasts. The IC50 values obtained were 90.5 and
96.3 µg/ml, respectively, and the results revealed that basil oil has potent
cytotoxicity.

Sharma et al. (2010) investigated the apoptosis inducing effect of

essential oil from aerial parts of O. viride in human colorectal
adenocarcinoma cells (COLO 205 cell line) by sulphorhodamine B (SRB)
assay. Cell cycle analysis and change in mitochondrial membrane
potential was quantified by flow cytometry. The results revealed that
adose and time-dependent inhibition by EO was determined with IC50
values of 0.070, 0.058 and 0.033 µl/ml at 24, 48, and 72 h, respectively.
This observed cell death was due to apoptosis, as established by annexin
V/PI assay, DNA ladder formation and scanning electron microscopy.
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296 RPMP Vol. 37: Essential Oils–II

The major constituents of essential oil were thymol (80.44%),

caryophyllene oxide (9.12%), a-pinene (2.23%), geranyl acetate (1.86%),
and 4-terpineol (1.86%), which constituted 95% of the essential oil.

The essential oils derived from O. sanctum showed the most

interesting anti-proliferative (cytotoxic) activity against MCF-7 (IC50
70.6-82.3 µg/ml) and LNCaP (IC50 70.0-75.0 µg/ml). The IC50 values
of O. gratissimum essential oils were 110.2-115.1 µg/ml and 105.3-
120.6 µg/ml on the cell proliferation of MCF-7 and LNCaP, respectively.
The essential oil extracted from O. basilicum was comparatively less
effective to inhibit cell growth of both the cell lines. IC50 values of O.
basilicum essential oil were found to be 260.3-270.7 µg/ml and 170.1-
172.1 µg/ml against MCF-7 and LNCaP cell lines. O. sanctum, O.
gratissimum and O. basilicum essential oils tested were less toxic
(IC50 149.9-395.3 µg/ml) against normal fibroblast (NIN-3T3) cell line
(Hussain, 2009).

The O. sanctum has been well known for its antioxidant property
with active ingredient such as eugenol and hence the plant has been
studied for its anticancer activity. The protective effect of alcoholic
extract of the leaves of O. sanctum on 3-methylcholanthrene (MCA),
7,12-dimethyl-benzanthracene (DMBA) and aflatoxin B, (AFB(1)
induced skin tumorigenesis in a mouse model was investigated. The
leaf extract of O. sanctum provides protection against chemical
carcinogenesis in one or more of the following mechanisms: (i) by acting
as an antioxidant; (ii) by modulating phaseI and II enzymes; (iii) by
exhibiting antiproliferative activity (Rastogi et al., 2007).

The essential oils isolated from the leaves of O. gratissimum were

tested for cytotoxic activity against P388 leukemia cells. The IC50 of
Cymbopogon oil was found to be 5.7 µg/ml while that of Ocimum oil
was 10.8 µg/ml. The mixture of the oils (1:1 v/v) showed an IC50 value of
10.2 µg/ml with no synergism in the cytotoxic activity (Dubey et al.,
1997).

Insect Repellent Activity

Oils from several Ocimum spp. have been shown to repel insects and
have larvicidal activities against house flies, blue bottle flies and
mosquitoes. Camphor, D-limonene, myrcene, thymol, are some of the
compounds in Ocimum oils that may provide the repellent properties.
Eugenol, citronellal and methylchavicol may responsible for the
larvicidal activity.
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Pharmacological Activities of Basil Oil: A Review 297

A study conducted by Mishra et al., (2012) investigated the repellent

activity of O. basilicum oil against two major economic important stored-
grain insect pests, red flour beetle Tribolium castaneum Herbst
(Coleoptera: Tenebrionidae) and rice weevil Sitophilus oryzae L.
(Coleoptera: Curculionide) adults. The results indicated that the
repellency of O. basilicum was 8.50 ± 0.22 for T. castaneum and
8.16±0.30 for S. oryzae. The repellency of both insect pests increased
with concentration from 0.05% to 0.40% at exposure time of 4 h.

O. gratissimum leaves from Cameroon are a potential source of essential

oil. Bioactivities were tested on the insect pest Sitophilus zeamais
(Motschulsky) (Coleoptera: Curculionidae), which is the major pest of stored
maize. Insecticidal activity was representatives of S. zeamais with 20 g of
maize grains powdered with various mixtures of essential oil and kaolin
(5 and 10%). The tested essential oils of O. gratissimum protected 74 % of
the test-material against the S. zeamais population after 4 days. A direct
application of the O. gratissimum on the test insects revealed 85.7% knock
down effect (Jirovetz et al., 2005). Similarly, Asawalam et al. (2008) assessed
the effect of essential oil of O. grattissimum leaves on S. zeamais for
repellency, mortality, progeny emergence and maize damage in the
laboratory. The oil was found to be moderately repellent to the maize weevil
and induced high mortality in the weevils. In addition, grains treated with
the essential oil showed significant reduction in the number of progeny
derived from surviving S. zeamais.

The dried ground leaves and essential oil of O. kilimandscharicum

in doses of 25.0 g leaves and 0.3 g essential oil per 250 g grain (maize or
sorghum) killed 100% of Sitophilus zeamais, Rhyzppertha dominica and
Sitotroga cerealella in 48 h. The best repellent activity was seen by 0.3
g essential oil/250 g grains against Sitophilus zeamais (Bekele &
Hassanali, 2001).

A study was carried out to determine the repellent activity of O.

gratissimum volatile oil against Simulium damnosum (black flies). A
12-month field study was conducted in three onchocerciasis endemic
communities (Idomido, Obiocamp, and Ikot Adaha) in Ini Local
Government Area of Akwa Ibom State, Nigeria. The results revealed
that topical application of 20% v/v concentration of the oil with liquid
paraffin as a base reduced the biting rate of S. damnosum by 90.2,
81.6, and 79.7%, in Idomido, Obiocamp, and Ikot Adaha respectively.
The oil gave protection against the bite of S. damnosum for at least 3 h.
A total of 710 adult S. damnosum were caught by individuals treated
with Ocimum oil, as against 4296 caught by the control group. (Usip et
al., 2006).
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Nematidical Activity

Asha et al. (2001) tested the essential oil of O. sanctum and its main
component eugenol in vitro in the Caenorhabditis elegans model.
Eugenol exhibited stronger anthelmintic effect (ED50 of 62.1 µg/ml) than
the essential oil (ED50 237.9 µg/ml). The essential oil of O. sanctum and
eugenol showed potent anthelmintic. Eugenol being the predominant
component of the essential oil, suggested as the potential candidate for
anthelmintic activity.

The ovicidal activity of the essential oil of O. gratissimum and its

main component eugenol was evaluated against Haemonchus contortus,
a gastrointestinal parasite of small ruminants. The oil and eugenol were
diluted in Tween 20 (0.5%) at five different concentrations. In the egg
hatch test, H. contortus eggs were obtained from the faeces of goats
experimentally infected. At 0.50% concentration, the essential oil and
eugenol showed a maximum eclodibility inhibition. These results
suggest a possible utilization of the essential oil of O. gratissimum as
an aid to the control of gastrointestinal helmintosis of small ruminants
(Pessoa et al., 2002).

Anti-Inflammatory and Immunomodulatory Properties

Anti-inflammatory activity of essential oil extract of O. sanctum leaf

(eugenol-enriched) was studied in Wistar rats by using carrageenan
induced hind paw edema method. The extract was administered 100
mg/kg body weight per intraperitoneally (i.p.) and the standard
paracetamol was also administered 5 mg/kg body weight per
intraperitoneally (i.p.). The extracted eugenol and paracetamol exhibited
significant (p< 0.05%) activity when compare with carrageenan control
(Thakur & Pitre, 2009). The essential oil extracted from O. gratissimum
inhibited the two enzymes, cyclooxygenase function of PGHS and
lipoxygenase L-1, with IC50 values, respectively, of 125 µg/ml and 144
µg/ml (Bedi Sahouo et al., 2003).

The oils extracted from fresh leaves (essential oil) and seeds (fixed
oil) of Tulsi have shown anti-inflammatory effects on experimental
animal’s hind paw edema induced by carrageenan, serotonin, histamine
and prostaglandin-E-2. The mechanism of action of the anti-
inflammatory effects of Tulsi could be the cyclooxygenase and
lipooxygenase pathways (Singh et al., 1996; Singh et al., 1997).

Fixed oils of various species of Ocimum viz O. sanctum, O. basilicum,

O. americanum, showed different response against phlogistic-agent-
induced paw edema. O. basilicum oils possess highest percentages of
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Pharmacological Activities of Basil Oil: A Review 299

linolenic acid (21.0%) and offered maximum inhibition of paw edema

(72.42%) (Singh, 1998). Fixed oil of Tulsi can inhibit enhanced vascular
permeability and leukocyte migration as evidenced by carrageenan
induced inflammatory stimulus (Singh, et al., 1996; Singh, et al., 2008).

In another study, (Mediratta et al., 2002) have investigated the

immunomodulatory effect of O. sanctum seed oil (OSSO) on some
immunological parameters in both non-stressed and stressed animals
and evaluated that OSSO appears to modulate both humoral and cell-
mediated immune responsiveness and these immunomodulatory effects
may be mediated by GABAnergic pathway.

Muscle Relexant and Analgesic

Experimental procedures using preparations obtained from O.

gratissimum were able to detect relaxant effect on isolated intestinal
smooth muscle (Madeira et al., 2002) and antinociceptive properties
(Aziba et al., 1999; Rabelo et al., 2003).

O. selloi essential oil (2, 20, and 200 mg/kg; p.o. orally) reduced, in a
dose-dependent way, the abdominal contraction induced by acetic acid
(0.6%; i.p.) and the diarrhea episodes induced by castor oil in mice
(Franca et al., 2008).

The antiulcerogenic activity of O. minimum essential oil was

demonstrated by reduction of gastric lesions, induced by indomethacin
or by ethanol, by 83.8% and 70.0%, respectively, at a dose of 0.125 ml/
kg in a mouse model (Maia et al., 2005). Similarly, fixed oil of O.
basilicum was found to possess significant antiulcer activity against
aspirin, indomethacin, alcohol, histamine, reserpine, serotonin and
stress-induced ulceration and secretion in pylorus ligated rats (Singh,
1999).

CNS Effect

Spontaneous sedative effect was detected in the thymol chemotype of

essential oil from O. gratissimum (Orafidiya et al., 2004). In another
study, the essential oil from O. minimum at doses of 1.5 mL/kg and 5.0
mL/kg, p.o., demonstrated CNS depressant activity in animals as
evidenced by observed changes in spontaneous motor activity, reactivity
to sound and touch (Maia et al., 2005). It has been suggested that the
anticonvulsant activity observed in the essential oil from O. gratissimum
extracted in the spring could be related to higher concentrations of
sesquiterpenes. (Löscher et al., 1988).
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The O. gratissimum oils obtained in each season were capable of

increasing the barbiturate-induced sleeping duration. The greatest effect
was obtained with the preparation from autumn, and the least effect
was observed with that from winter, which was not active in the lesser
dose administered. Eugenol was the most abundant compound in the
essential oil from each season, with the greatest relative percentage
detected in autumn (56.10%) (Cristiana et al., 2006).

Antiprotozoal Activity

A study carried out by Oliveira et al. (2009) revealed that essential oils
of O. gratissimum inhibited the growth of L. chagasi promastigotes in a
dose-dependent way, with IC50 75 µg/ml. Similarly, the eugenol-rich
essential oil of O. gratissimum progressively inhibited Leishmania
amazonensis growth at concentrations ranging from 100 to 1000 µg/ml.
The IC50 of the essential oil for promastigotes and amastigotes were,
respectively, 135 and 100 µg/ml and the IC50 of eugenol was 80 µg/ml
for promastigote forms (Ueda-Nakamura et al., 2006). In another study,
essential oils from O. basilicum and O. sanctum grown in Mississippi
showed in vitro activity against Leishmania donovani (IC50=37.3-49.6
µg/ml), which was comparable to the activity of basil oil (IC50 = 40-50
µg/ml). Minor basil oil constituents (+)-d-cadinene, d-3-carene, a-
humulene, citral, and (-)-(E)-caryophyllene had antileishmanial activity
(Zheljazkov et al., 2008).

Oil of O. gratissimum showed significant antimalarial activities

against P. berghei in the four-day suppressive in vivo test in mice. At
concentrations of 200, 300 and 500 mg/kg of mouse per day, the essential
oil of O. gratissimum showed 55.0%, 75.2% and 77.8% supression,
respectively. Chloroquine (10 mg/kg of mouse, positive control) had a
suppressive activity of 100% (Tchoumbougnang et al., 2005). At
concentrations from 20 to 250 µg/ml, the essential oil of O. gratissimum
progressively inhibited the protozoan growth of Herpetomonas
samuelpessoai, a nonpathogenic trypanosomatid. The IC50, in defined
and complex media, at 28 C were 100 and 91 µg/ml, respectively (Holetz
et al., 2003).

In another study, essential oil of O. basilicum (2 mg/ml) and its

purified substances demonstrated antigiardial activity. Linalool (300
µg/ml), however, was able to kill 100% parasites after 1 h of incubation,
which demonstrates its high antigiardial potential (de Almeida et al.,
2007).
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Pharmacological Activities of Basil Oil: A Review 301

AChE and BChE Inhibitors

Essential oil of O. sanctum oil showed moderate AChE inhibition with

IC 50 value of 1.6 mg/ml. The oil consisted mainly of only three
components: eugenol (59%), â-caryophyllene (33%), and á-humulene
(3.0%) (Dohi et al., 2009). In another study, at concentrations of 1 mg/
ml, essential oils of both green and purple varieties of O. basilicum,
had little inhibitory activities (10.7%, 33.3%), whereas they remarkably
inhibited BChE with 74.1% and 79.1% inhibition, respectively (Orhan
et al., 2008).

Human Studies

A recent study tested a range of concentrations of O. gratissimum oil in

comparison to 10% benzoyl peroxide anda placebo, over a period of 4
weeks, for the reduction of acnelesions in a population consisting mainly
of students. O. gratissimum oil was incorporated at concentrations of
0.5%, 1%, 2% and 5% v/v in four different bases (polysorbate 80,
cetomacrogol, petrolatum and alcohol) resulting in 16 parallel
experimental groups. Preparations containing 2% and 5% Ocimum oil
in alcohol and 5% in cetomacrogol were significantly more active than
benzoyl peroxide (P < 0.05), The most active 5% preparations produced
skin irritation but the authors considered a 2% preparation in
cetomacrogol to be suitable for the management of acne (Orafidiya et
al., 2002). Another clinical study was carried out by the same author
using a combination of O. gratissimum and Aloe vera gel. It was found
that Aloe vera gel enhanced the antiacne properties of Ocimum oil. The
oil or its combination with Aloe vera gel was found to be more effective
than 1% clindamycin in the treatment of Acne vulgaris (Orafidiya et
al., 2004). In another study conducted by Pansanga et al. (2010), it was
found that a 3% sweet basil microemulsion should be safe and well
tolerated on male human skin.

CONCLUSIONS

This review attempts to shed light on the ethnopharmacology,

phytochemistry, pharmacology and few clinical studies of basil oil i.e.
EOs extracted from different species of Ocimum. The results reviewed
in this article are aimed at attracting the attention of researchers
seeking new drugs from natural products as well as those investigating
the pharmaceutical diversity of essential oils. The data presented
provide also a basis for reviving the old art of ‘Ocimum oil therapy’
based on our modern scientific knowledge of their mode of action, and
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302 RPMP Vol. 37: Essential Oils–II

it can be said that Ocimum essential oils have shown very promising
results extending from their activity as antimicrobial, antioxidant and
cytotoxic agents to antiprotozoal, nematidical and analgesic. Moreover,
insecticides and insect repellent agents based on Ocimum essential oils
or their constituents have demonstrated efficacy against a range of
stored product pests, domestic pests. Because of their volatile nature,
there is a much lower level of risk to the environment than with current
synthetic pesticides. Thus Ocimum essential oils and their constituents
can hopefully be considered in the future for more clinical evaluations
and possible applications, and as adjuvants to current medications.

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