2011

NPC Natural Product Communications Vol. 6

No. 10
Antibacterial Activity of the Essential Oils of Pistacia 1505 - 1506
lentiscus Used in Moroccan Folkloric Medicine
Fatima Zohra Mharti, Badiaa Lyoussi and Abdelfattah Abdellaoui

Department of Biology, University Sidi Mohamed Ben Abdellah, Faculty of Sciences Dhar Mehraz,
Laboratory of Physiology Pharmacology and Environmental Health, USMSA- B.P. 1796. Atlas,
Fez –Morocco

abdellaouia@yahoo.fr

Received: May 31st, 2011; Accepted: June 17th, 2011

The essential oil of the leaves of Pistacia lentiscus, collected from the middle Atlas in Morocco, was analyzed by GC and GC–MS.
Altogether 43 components in concentrations of more than 0.2% were identified representing 97.4% of the oil composition. The
main constituents were germanicol (12.8%), thunbergol (8.8%), himachalene (7.4%), trans-squalene (6.7%), terpinyl propionate (6.7%),
3,3-dimenthol (6.2%) and cadina-1.4-diene (5.1%).The oils showed strong activity against Klebsiella pneumonia, but no activity against
Pseudomonas aeruginosa.

Keywords: Antibacterial activity, Pistacia lentiscus, essential oil, nosocomial infection, germanicol, GC/MS.

The aim of this work was to determine the composition of Table 1: Composition of the essential oil from leaves of Pistacia
lentiscus (%).
the leaf essential oil of Pistacia lentiscus L. (family
Anacardiaceae) from Morocco and to evaluate its Compounds (%) RI Compounds (%) RI
antibacterial activity against two Gram-negative bacteria 3-Carene 0.9 910 Pinane, 2,3-epoxide 0.2 1103
responsible for nosocomial infections encountered in the Terpinyl propionate 6.7 931 ɑ–Cubebene 1.1 1451
p-Metha-1.8-diene 2.9 951 Longifolene 1.8 1377
University Centre Hospital of Fez (Morocco). Thunbergol 8.8 1413 Copaene 0.3 1425
trans-Squalene 6.7 1785 Ylangene 0.2 1431
The leaf essential oil of P. lentiscus from Morocco was α-Phellandrene 0.3 1031 Himachalene 7.4 1505
yellow in color and was obtained in a yield of 0.2%. This Isopseudocumenol 2,0 975 α–Guaiene 2.4 1515
was higher than those in Algeria and Italy (0.02-0.12% and α–Pinène 0.2 1049 Himachala-2,4-diene 3.7 1486
0.09-0.32%, respectively) [1,2]. Forty-three components 3,3-Dimenthol 6.2 1586 Germanicol 12.8 1632
cis-Ocimene 0.5 1062 Isoledene 1.9 1499
were identified, representing 97.4% of the total oil
1R-ɑ-Pinene 2.9 962 Cadina-1.4-diene 5.1 1511
(Table 1). This percentage is higher than those reported for Caprylone 0.3 1236 Epizonarene 0.4 1156
P. lentiscus from Tunisia and Greece (58% and 13.5%, Pseudocumenol 2.7 985 Carnegine 0.5 1409
respectively) [3,4]. The main component of the essential Camphor 0.6 1077 Globulol 0.7 1550
Borneol 0.5 1159 Caryophyllene oxide 0.3 1575
oils was germanicol (12.8%). Other major constituents Isoborneol 3.6 1089 Agarospirol 0.3 1581
were thunbergol (8.8%), himachalene (7.4%), trans- trans-ɑ-Terpinyl 1.3 1349 Carotol 0.2 1499
squalene (6.7%), terpinyl propionate (6.7%), 3,3 dimenthol butanoate
(6.2%) and cadina-1.4-diene (5.1%). Hexenyl valerate 0.2 1131 Muurolol 2.8 1530
Isopentyl hexanoate 0.3 1142 Patchoulene 2.5 1632
α–Truxene 1.7 1275 Ledene oxide-(II) 0.2 1687
The antimicrobial test results are presented in Table 2. 2,5-Dibutyfuran 3.6 1016 Benzyl benzoate 0.2 1716
Strong activity was observed against the Gram-negative Furanone 0.2 893 Total 97.4
Klebsiella pneumonia, with an inhibition zone of 16 mm, Abbreviations: Retention index (RI); Area (%).
but no activity was observed against Pseudomonas
aeruginosa, which is also resistant to most standards Table 2: Antimicrobial activity of Pistacia lentiscus leaf essential oil.
antibiotics, except Imipenem (Table 2). This drug is Test microorganism Inhibition Antibiotic standards
typically used for the treatment of hospitalised patients zone (mm)
Klebsiella pneumoniae 16 35 (IMP), 30 (AK), 0 (LEV),
with nosocomial infection in order to avoid bacterial 0 (AMP)
resistance. Based on these results, it is possible to conclude Pseudomonas aeruginosa n/a 44 (IMP), 0 (AK), 0 (LEV),
that the essential oil of Moroccan P. lentiscus has a 0 (CIP), 0 (AMX)
Abbreviations Standard antibiotic disks: Imipenem IMP, Amikacin AK,
stronger antibacterial activity than standard antibiotics Levofloxacin LEV, Ciprofloxacin CIP, Ampicillin AMP, Amoxicillin AMX, n/a:
such as levofloxacin and ampicillin (Table 2). not active
1506 Natural Product Communications Vol. 6 (10) 2011 Mharti et al.

Experimental
Plant material: Fresh leaves of Pistacia lentiscus L. were
collected in January 2009 in a winery located in hills in the
region of Berkin, 100 km from Gerssif city, Morocco. The
leaves were dried for 7 to 10 days in the shade, then stored
in cloth bags at 5°C until extraction.
Preparation and analysis of essential oil: Air-dried leaves
(100 g) were subjected to hydrodistillation for 3 h with 500
mL distilled water using a Clevenger-type apparatus,
according to the European Pharmacopoeia [5]. The oil
obtained was collected and dried over anhydrous sodium
sulfate and stored in a refrigerator at 4-5°C prior to
analysis. Oil yield was based on dry weight.
The volatile constituents were analyzed on a Thermo
Fischer capillary gas chromatograph directly coupled to a
mass spectrometer system (model GC ULTRA S/N
20062969; Polaris QS/N 210729), HP-5MS non polar
fused silica capillary column (60 m x 0.32 mm, 0.25 μm
film thickness). The oven temperature was maintained at
40°C for 2 min, then increased at a programmed rate of
2°C/min to a final temperature of 260°C, which was
maintained for 10 min; injector temperature 250°C. The
carrier gas was helium at a flow rate of 1mL/min. Samples
were run in n-hexane with a dilution ratio of 1:10.
Compounds were identified by matching their MS and
retention index with those reported in the literature [6].
The volume of injected specimen was 1 μL of diluted oil;
split injection technique; ionization energy 70 eV in the
electronic ionization mode; ion source temperature 200°C,
scan mass range m/z 40-650, and interface line temperature
300°C. The essential oil components were identified by
comparing their retention times and mass fragmentations
with the NIST-MS and by literature comparison [7].
Antimicrobial activity assessment: The microorganisms
tested were the Gram-negative bacteria Klebsiella
pneumonia and Pseudomonas aeruginosa. These were
isolated in a hospital environment from clinical patients in
reanimation service (CHU, Morocco).
For the susceptibility screening test, an agar-disc-diffusion
procedure was adapted from a method used earlier [8].
Each microorganism stock was suspended in Mueller-
Hinton broth and incubated at 37°C for 18–24 h. The
overnight cultures were diluted and adjusted in order to get
a density of 108 c.f.u./mL (0.5 McFarland turbidity
standard). These were flood-inoculated onto the surface of
MH agar plates and 6 mm diameter, sterile filter discs of
Whatman paper N3, impregnated with 15 µg/disc of the
essential oil, were delivered onto the inoculated agar
(MH). The plates were incubated for 18 h at 35°C.
Antimicrobial activity was evaluated by measuring the
zones of inhibition. The standards used were Imipenem,
Amikacin, Levofloxacin, Ciprofloxacin, Ampicillin and
Amoxicillin. The tests were carried out in duplicates.
Acknowledgments - The authors would like to thank the
Regional Center of Interface, University Sidi Mohamed
Ben Abdellah, Fez, Morocco for the gas chromatography
coupled with mass spectrometry (GC/MS).

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