T.

mascatenses
J. Essent. Oil Res., 18, 465-468 (July/August 2006)

The Composition and Antimicrobial Activity of Leaf

Essential Oil of Teucrium mascatenses Boiss. from
Oman

Abdulkhader Hisham,* Nirmal Pathare, Salim Al-Saidi and Ahmed Al-Salmi

Departments of Chemistry and Biology, College of Science, P.O.Box-36, Sultan Qaboos University, Al-Khode-123, Oman

Abstract
A hydrodistilled oil from the leaves of Teucrium mascatense Boiss. was analyzed by GC/MS. Twenty-one com-
ponents amounting to 91.2% of the oil were identified with linalool (27.8%), linalyl acetate (12.6%) and β-eudesmol
(10.1%) being the major constituents of the oil. The antimicrobial activity of the oil was tested against a panel of 17
bacterial and six fungal strains by the disc diffusion method. The oil inhibited the growth of all test organisms at vari-
ous levels. The minimal inhibitory concentrations (MIC) were also determined.

Key Word Index

Teucrium mascatensis , Lamiaceae, essential oil composition, linalool, linalyl acetate, β-eudesmol, antimicrobial
activity.

Introduction
The genus Teucrium (Lamiaceae) is represented by about
300 species (1), of which 49 species are endemic to Europe
mainly in the Mediterranean region and many of these species
are used in the folk medicine of many countries as stimulants,
tonics, stomach ache remedy and also as anti-diabetic agents
(2). The Teucrium species are considered as potential source
of neo-clerodane diterpenoids and they are rich in essential
oils as well (2).
Even though, several reports deal with the essential oil
composition of more than thirty Teucrium species, the studies
on the species growing in this region is rather limited except for
T. stocksianum (3,4) , T. melissoides (5) , T. flavum (6) and T.
polium (7). In general, the essential oil chemistry of Teucrium
dominates with sesquiterpe contents particularly compounds
such as β-caryophyllene and germacrene D.
Teucrium mascatensis Boiss. is an aromatic perennial, a
native plant of Oman, which grows 25 cm high and frequently
seen on rocky hill and mountain slopes in northern Oman (8).
It shares the common Arabic name ja’dah with a related spe-
cies T. stocksianum Boiss., a plant more common throughout
the Middle East and is popular for its use in folk medicine as
a fever remedy. Teucrium mascatense is also used as a fever
remedy as well to reduce the blood flow during menstruation.
This Teucrium species is morphologically quite distinct from
T. stocksianum and has neither been chemically nor pharma-
cologically investigated so far.
*Address for correspondence
1041-2905/06/0004-0465$14.00/4­—© 2006 Allured Publishing Corp.
Experimental
Plant material used in this study was collected from the
higher planes of ‘green mountain’ (Jabal Al-Akdhar in Arabic),
the main mountain massif of northern Oman, situated 2800 m
above the sea level, in April 2004 and authenticated by Annette
Patzalt, Department of Botany, SQ university, Oman where a
voucher specimen has been deposited.
Oil isolation: A sample of pleasant smelling genuine oil
was isolated by hydrodistillation of 50 g shade-dried powdered
aerial parts of T. mascatense using a Clevenger-type apparatus
for four hrs (yield 0.8%).
Preliminary GC analysis: The preliminary GC analysis
was carried out
by means of a HP 5890 gas chromatograph equipped with a
HP-1-cross-linked methylsilicone gum capillary column (25 m
x 0.32 mm x 0.52 µm film thickness), using a FID detector.
GC/MS analysis: The oil was diluted with an appropri-
ate volume of dichloromethane and analyzed by GC/MS on a
Shimadzu model (GC-MS-QP/5050A) instrument equipped
with HP-5 (5%-phenyl)-methylpolysiloxane-nonpolar-capillary
column (30 m x 0.32 mm x 1.0 µm thickness) and interfaced
with a quadrupole mass spectrometer.
Analytical conditions: The injector and interface tem-
peratures were kept at 275°C and 300°C, respectively. The
oven temperature was programmed from 70°-270°C at a rate
of 3°C/min. Helium was used as the carrier gas with a linear
velocity of 74.6 cm/s and the total flow rate was 39.9 mL/min.
Received: December 2004
Revised: June 2005
Accepted: June 2005

Vol. 18, July/August 2006 Journal of Essential Oil Research/465

 Hisham et al.
Table I. Percentage composition of the leaf oil of Teucrium
mascatense
Compound %
Monoterpenes
α-pinene 0.5
β-pinene 0.5
limonene 5.7
cis-linalool oxide (furanoid) 4.6
trans-linalool oxide (furanoid) 4.1
linalool 27.8
verbenol 1.3
p-mentha-1,5-dien-8-ol 1.2
α-terpineol 3.0
myrtenal 0.8
verbenone 1.1
trans-carveol 1.0
carvone 2.8
linalyl acetate 12.6
α-terpinyl acetate 0.9
neryl acetate 1.0
geranyl acetate 1.5
Sesquiterpenes the oil were identified (see Table I ) . Eighteen monoterpenes
α-bergamotene 5.0
β-selinene 2.5
caryophyllene oxide 2.9
β-eudesmol 10.1
correct isomer not identified
*
Mass spectra were continuously recorded from 40 to 500 m/z.
The MS operating parameters were: ionization voltage 70 eV,
scan rate 500 amu/s. The components of the essential oil were
identified by comparison of their mass spectral data with the
reference spectra (Wiley 229, 2000) in the data base.
Microbial cultures growth conditions: Test microorgan-
isms included the following standard antimicrobial susceptibility
strains: Staphylococcus aureus (NCTC 6571), Escherichia coli
(NCTC10418), Pseudomonas aeruginosa (NCTC10662) besides
a battery of Gram positive and Gram negative strains given in
Table II as per the NCCLS guidelines (9). Cultures of these
bacteria were grown in Mueller-Hinton broth (Difco) at 37°C
and maintained on slopes of nutrient agar (Difco) at 4°C.
Antimicrobial activity assay: The oil was tested for an-
timicrobial activity using the disc diffusion technique on solid
media as described by Bauer et al (10). Sterile, 6-mm diameter
Whatman 41 discs [containing filter sterilized oil initially diluted
4 mg/mL with ethylene glycol and further diluted to achieve
required disc concentrations with phosphate buffered saline
(w/v)] were placed on plates of Mueller-Hinton agar (Difco),
which had been surface spread with 0.1 mL of logarithmic
phase bacteria at a density adjusted to a 0.5 McFarland turbid-
ity standard (108 colony-forming units [CFU]/mL). The plates
were then incubated for 24 h at 37°C. Disc containing 0.04
mL ethylene glycol was placed in each plate as control. The
results were recorded by measuring the average zones of growth
inhibition surrounding the discs. Standard antibiotic discs as
per the requirement of ‘Laboratory Control and Antimicrobial
Therapy’(11) were used in parallel. Antifungal disc diffusion
assay was performed by employing above techniques utilizing
466/Journal of Essential Oil Research Vol. 18, July/August 2006
Sabouraud’s agar and 0.2 mL culture suspension. The plates
were incubated at 27°C for 48 h.
Minimal inhibitory concentration (MIC): The oil was
tested for antibacterial activity using the microbroth dilution
method in broth media Mueller-Hinton (Difco) as per NCCLS
guidelines (12). In these experiments, 50 µL of a suspension
containing 1 x× 106 CFU/mL was added to 100 µL of susceptibil-
ity test broth containing serial twofold dilutions of the essential
oil in sterile ELISA plate wells. All plates were incubated at
37°C for 24 h before being read. The MIC was considered
the lowest concentration of the sample that prevented visible
growth. Minimum bactericidal concentrations (MBCs) were
determined by subculturing, 10 µL from each negative well
and from the positive growth control. MBCs were defined as
the lowest concentration yielding negative subcultures. All
samples were examined in duplicate.
Results and Discussion
The chromatogram of T. mascatense leaf oil revealed that
25 peaks were detected of which 21 representing 91.4 % of
amounting to 70.9 % in the oil of which 17 were identified
except for a minor component (0.6 %). Linalool (27.8 %),
linalyl acetate (12.6 %) were predominant compounds in the
monoterpene series. Seven sesquiterpenes amounting to 29.2
% were detected of which four compounds, β-eudesmol (10.1
%), α- bergamotene (5.0 %), caryophyllene oxide (2.9 %) and
β-selinene (2.5 %) were identified. The other compounds could
not be identified due to the lack of reference spectra in the
library. Teucrium mascatense leaf oil was found to be rich in
oxygenated monoterpenes. The composition of T. mascatense
oil was found to be quite different from that of T. stocksianum
reported from the United Arab Emirates by Al Yousuf et al.(3)
and that from Iran by Mojab et al. (4) in the number as well as in
the nature of components. The oil of T .stocksianum from UAE
contained more than 70 components in which α-cadinol and
δ-cadinene were the major ones, whereas α-pinene, β-pinene
and β-cubebene were the major components in the oil from
Iran. In addition, the oil composition of T. mascatense seems
to be different from that reported for seven other Teucrium
species by Kovacevic et al. (13) in which sesquiterpenes were
found to be the major components in the oil. But T. mascatense
oil partially resembled the oil of T. montana (13) in terms of
its high percentage of β-eudesmol and also that of T. creticum
(14) leaf oil in its high percentage of linalool.
In the present study, the oil from T. mascatense was found
to exhibit broad spectrum antimicrobial activity. Antimicrobial
activity was tested using a battery of Gram positive and Gram
negative bacteria as well as fungi and molds, along with inter-
national standard susceptibility strains S. aureus [NCTC6571],
E. coli [NCTC10418] and P.aeruginosa [NCTC10662]. Most
of the Gram positive strains of saprophytic species as well as
pathogenic species of Staphylococci and Streptococci exhibited
increased susceptibility to the constituents of this oil. Gram
negative organisms were also found to be sensitive to the oil,
with higher susceptibility being exhibited by enteric strains
belonging to E. coli, P. aeruginosa and Salmonella species.
The oil from T. mascatense retained its activity up to 0.2 mg
 T. mascatenses

Table II. Antimicrobial activity of the leaf oil of Teucrium mascatense by the disc diffusion method/MIC

Diameter of zone of inhibition [mm] MIC

Bacterial Strains Amox-Clav Cefotaxime Tetracycline Oil oil

Gram-Positive [20 µg] [30 µg] [30 µg] [0.4 mg] [0.2 mg] [mg/mL]
# S. aureus* [NCTC6571] 13.0 6.5 9.0 20.0 17.5 2.0
◘S. aureus * 12.0 6.5 8.0 17.0 15.0 2.5
◊S. albus * 14.0 6.0 9.0 20.0 17.5 2.0
◘S. epidermidis * 11.0 6.0 9.0 18.5 16.0 2.0
◘Strept. mitis** 14.0 8.0 10.0 20.0 17.5 1.5
◊Strept. sanguis** 16.0 8.0 10.0 19.0 17.0 1.5
◊M. luteus*** 10.0 6.0 9.0 11.0 9.5 2.5
◊B. subtilis **** 7.0 2.0 11.0 17.5 15.5 2.0
◊B. cereus **** 6.0 2.0 9.0 18.0 16.5 1.5

Gram-Negative
#E. coli■ [NCTC10418] 6.5 10.0 6.5 15.0 13.5 6.0
◘E. coli■ 7.5 10.0 5.5 11.0 9.5 7.0
◊Entero. aerogenes■ ■ 3.5 5.5 5.0 5.5 3.5 8.0
◊Kleb. pneumoniae■■■ 2.5 6.5 4.0 9.0 7.5 8.5
◊Salmonella typhi 8.0 11.0 7.5 13.5 11.5 6.5
◊Proteus vulgaris 6.0 10.5 2.0 8.5 7.0 8.5
#Ps. aeruginosa■■■ ■
[NCTC10662] 0.0 2.5 0.0 11.5 9.5 8.0
◊Ps. aeruginosa ■■■■ 0.0 2.0 0.0 8.5 06.5 8.5

Fungal Strains Diameter of zone of inhibition [mm] Mycostatin

1.6 mg 0.8 mg 0.4 mg 0.2 mg 15 µg/mL
◊Candida albicans 5.0 3.5 2.0 1.0 5.5
◊Saccharomyces cerevisiae 5.5 4.5 2.5 1.0 6.0
◊Aspergillus flavus 3.5 2.5 1.0 0.0 10.0
◊Rhizopus stolonifer 5.0 3.5 2.0 1.0 4.5
◊Penicillium notatum 6.5 4.5 3.0 1.5 8.0
◊Fusarium oxysporum 4.5 3.0 2.0 1.0 4.0
N. B.-zone of inhibition values are the average of four determinations in four different directions; Whatman 41 (6 mm) discs were soaked with each sample tested at the given
concentration in phosphate buffered saline (w/v); Initial stock dilution was done in ethylene glycol, the control disc containing ethylene glycol did not exhibit any antibacterial/
antifungal activity
Amox-Clav- Amoxicillin- Clavulanate; bacterial strains : S.* -Staphylococcus, Strept. **- Streptococcus, M. ***- Micrococcus, B. ****-Bacillus, E. - Escherichia, Ent. - Enterobacter,
K. - Klebsiella, Ps.- Pseudomonas; #- the standard NCTC strains recommended for standardization of antibiotic susceptibility test; - wild type strains isolated from human
normal flora; ◊- wild type strains isolated from environment

Blunden, The composition of the essential oil of Teucrium stocksianum

concentration in the disc diffusion assay. The MIC range varied
from 1.5-2.5 mg/mL for Gram positive organisms and 6.0-8.5
mg/mL for Gram negative organisms. The oil also showed activ-
ity predominantly against yeasts and moderate activity against
molds; however the inhibitory oil concentrations necessary for
these were higher compared to the various bacterial species
tested. Our observation suggests that the oil from T. mascatense
is effective against Gram positive cocci and yeasts which typi-
cally form the predominant opportunistic skin microflora; and
this may be reflected in its local usage.
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