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Chemical Composition and Antibacterial Activity of

Essential Oil of Myrtus communis L. Growing Wild in
Italy and Turkey
a a a a
Felice Senatore , Carmen Formisano , Francesco Napolitano , Daniela Rigano & Musa
b
Özcan
a
Dipartimento di Chimica delle Sostanze Naturali, Università degli Studi di Napoli “Federico
II”, Via D. Montesano, 49, I-80131, Napoli, Italy
b
Department of Food Engineering, Faculty of Agriculture, Selçuk University, Konya, 42031,
Turkey
Published online: 12 Mar 2013.

To cite this article: Felice Senatore , Carmen Formisano , Francesco Napolitano , Daniela Rigano & Musa zcan (2006)
Chemical Composition and Antibacterial Activity of Essential Oil of Myrtus communis L. Growing Wild in Italy and Turkey,
Journal of Essential Oil Bearing Plants, 9:2, 162-169, DOI: 10.1080/0972060X.2006.10643489

To link to this article: http://dx.doi.org/10.1080/0972060X.2006.10643489

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 Jeobp 9 (2) 2006 pp 162 - 169 162

ISSN 0972-060X

Chemical Composition and Antibacterial Activity of Essential Oil of

Myrtus communis L. Growing Wild in Italy and Turkey

Felice Senatore1,* , Carmen Formisano1 , Francesco Napolitano1 ,

Daniela Rigano1 , Musa Özcan2
Downloaded by [East Carolina University] at 16:50 23 August 2013

1
Dipartimento di Chimica delle Sostanze Naturali, Università degli Studi di Napoli
“Federico II”, Via D. Montesano, 49, I-80131 Napoli, Italy.
2
Department of Food Engineering, Faculty of Agriculture, Selçuk University,
Konya 42031, Turkey.
Received 15 May 2005; accepted 22 July 2006

Abstract: The composition of the essential oils from aerial parts of Myrtus communis L.
(Myrtaceae) from Italy and Turkey has been studied. The oil content was 0.33% and 0.38% (v/w), on
a dry weight basis, respectively for the Italian and Turkish sample. The oil compositions were ana-
lyzed by GC and GC/MS. Altogether 39 compounds were identified accounting for 94.8% and 95.9% of
the oil, respectively for the Italian and Turkish oil. Italian oil (I) was characterized by α-pinene (15.7%)
and 1,8-cineole (16.5%). Linalool (36.5%) and linalyl acetate (16.3%) were the most abundant compo-
nent in the Turkish oil (T). The oils showed antibacterial activity mostly against Gram + bacteria; the
Turkish oil was more active compared with the Italian oil, probably for a major presence of active
compounds such as linalool, α-terpineol and geranyl acetate.

Key words: Myrtus communis L.; essential oil; linalool; linalyl acetate; 1,8-cineole;
antibacterial activity.

Introduction: Myrtle (Myrtus communis L., Myrtaceae) is an evergreen shrub or a

small tree with white, fragrant flowers. It grows spontaneously through the Mediterranean
area, where it is also cultivated as an ornamental plant. In Italy the plant is known as mortella
and spreads over the Mediterranean coasts1. In Turkey, myrtle is called hambeles, mersin
or murt and it is found growing in pine forests and riversides, particularly in the Taurus
mountains2, from just above sea level to 500-600 m. This plant is known for culinary pur-
poses. The leaves are strongly aromatic and they are commonly known for the presence of
essential oils, whose composition determines the specific aroma of plants and the flavour of
the condiment. The oils extracted by steam distillation of fruits are used in flavour and

*Corresponding author: (Felice Senatore)

E-mail: <fesenato@unina.it>
 Felice Senatore et al. / Jeobp 9 (2) 2006 pp 162 - 169 163

fragrance industries3,4. The fruits, considered a rich source of tannins5, are very astringent
and are used as a condiment, as a substitute for pepper. In Turkey its fresh and/or dried
leaves oils are used in cosmetics sauces, confectionery and beverage industry6-11. This spe-
cies is also widely used in Italian folk medicine because of its astringent and balsamic prop-
erties12,13. Leaves and fruits decoctions or infusions of this plant are used as stomachic,
hypoglycemic, antimicrobic, appetizing, antihemorrhagic, for cough and oral diseases, consti-
pation and externally for wound healing12,14-16. A survey of the literature reveals a number of
studies concerning the chemical composition of myrtle oil also according their provenance4,8,17-
26
. A previous study11 on Turkish myrtle leaves essential oil, characterized by the presence of
myrtenyl acetate ranging between 14.5 and 10.8%, reports linalool (18.6-16.3%) and 1,8-
cineole (18.2-10-5%) as the main components of the oil. Essential oils obtained from the
leaves of plants growing in Liguria (Italy) is characterized by more than 90% of monoterpe-
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nes24 with α-pinene (28.9-41,6%) and 1,8-cineole (24.2-25.5%) as the main components of
the oil.
The oil from leaves of plants grown in Sardinia (Italy)26 also contains α-pinene (30.0-
59-5%) and 1,8-cineole (15.9-41.7%) as the most abundant components. Spanish oil8,20 is
characterized by a high content of myrtenyl acetate (35.9%) and a low content of α-pinene
(8.2%), while Tunisian oil27 shows α-pinene (52.9%) and 1,8-cineole (18.97%) as the main
components with myrtenyl acetate present in very low amount (0.16%); the same was
found for the oil from Corsica21 with α-pinene (53.5-56.7%) and 1,8-cineole (18.85%) as
the main components, while myrtenyl acetate represents only 0.82% of the oil. Chalchat et
al.21 compared also the chemical composition of 15 commercial samples of essential oil
from leaves of myrtle grown in other different Mediterranean countries and pointed out that
the differentiation among these oils can be made respect to the content of α-pinene and
myrtenyl acetate. The oil from Greece8 was found rich in linalyl acetate (31.4%), limonene
(21.8%) and α-pinene (18.0%). Recently28 we have demonstrated the inhibitory effect of
the essential oil from myrtle collected in Turkey on the mycelia growth of Agaricus campestris
(Lange) Sing. As a continuation of our studies, we have determined the chemical profile of
the myrtle oil obtained from plants collected in Maiori (Amalfitan coast, Campania, Italy)
and Büyükeceli-Gülnar-Mersin (Içel, oriental Mediterranean sea, Turkey) and we have evalu-
ated their antimicrobial activity, to further validate the medicinal properties of the plant.

Experimental
Plant material: Aerial parts of Myrtus communis L. were collected in June 2005, in
Campania (on the Lattari mountains, Amalfitan coast) and near Içel (Buyukeceli/Gulnar),
South Turkey. Plant material was identified by F. Senatore (Italian sample) and at the De-
partment of Biology, Faculty of Science and Education, Selçuk University, Turkey (Turkish
sample). Voucher specimens (NAP # 10 and NAP # 11) have been deposited in the Her-
barium Neapolitanum (NAP), Dipartimento di Biologia Vegetale, Università degli Studi di
Napoli “Federico II”, Italy.

Isolation of the essential oil: Dried aerial parts of the plants (leaves and flowers,
 Felice Senatore et al. / Jeobp 9 (2) 2006 pp 162 - 169 164

25 g), were collected and cut in small pieces and then subjected to hydrodistillation for 3 h,
with a Clevenger apparatus according to the standard procedure reported in the European
Pharmacopoeia29. The dry material yielded a 0.33% (Italian oil, I) and 0.38% (Turkish oil,
T) of pale yellow oil having a typical, strongly aromatic and pleasant-smell.

Gas Chromatography (GC): The oils were analyzed using a Perkin-Elmer Sigma-
115 gas chromatograph, equipped with a FID and a data handling system. Analytical condi-
tions were: DB 5 fused-silica capillary column (30 m x 0.25 mm, film thickness 0.25 µm),
carrier gas, He. Column temperature: 40°C, with 5 min initial hold, and then to 260°C at 2°C/
min, 260°C (20 min) using He as the carrier gas (1.0 mL/min). 1 µL of sample (1:100 n-
pentane solution) was injected in split mode (1:20). Retention indices (Ri) were determined
in relation to a homologous series of n-alkanes (C8-C24) under the same conditions. Compo-
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nents relative concentrations were obtained by peak area normalization. The percentage
composition of the essential oils was calculated from GC peak areas without using correc-
tion factors.

Gas Chromatography-Mass Spectrometry (GC-MS): GC/MS were performed

on a Agilent 6850 Ser. II apparatus equipped with a HP 5 fused-silica capillary column (30 m
x 0.25 mm; film thickness 0.33 µm), coupled with a Agilent Mass Selective Detector MSD
5973. The temperature program used was the same as described above. The MS was
operated in EI mode with ionization voltage 70 eV, electron multiplier energy 2000 V. The
oven programming was the same as above; transfer line was kept at 295°C.

Components identification: The oil components were identified from their GC re-
tention indices30, by computer matching against commercial (NIST 02, Wiley-275) and home-
made library mass spectra made up of pure substances and components of known oils, as
well as MS literature data31,32 and, whenever possible, by co-injections with authentic com-
pounds available in our laboratories.

Evaluation of antimicrobial activity: The oils antibacterial activity was evaluated

by determining the minimum inhibitory concentration (MIC) using the broth dilution method33.
Eight bacteria species [Bacillus cereus (ATCC 11778), Bacillus subtilis (ATCC 6633),
Staphylococcus aureus (ATCC 25923), Streptococcus faecalis (ATTC 29212), Escheri-
chia coli (ATCC 25922), Proteus mirabilis (ATCC 25933), Pseudomonas aeruginosa
(ATCC 27853), Salmonella typhi Ty2 (ATCC 19430)], selected as representative of the
class of Gram + and Gram -, were tested as previously described34.

Results and Discussion: The results of this study are presented in Table 1 where
the identified compounds are ordered with respect to their elution times on a DB 5 capillary
column. The analysis of the myrtle essential oil has allowed the identification of thirty three
and twenty-six compounds, representing the 94.8% (I) and the 95.9% (T) of the total oil,
respectively. From the results it appeared that the total amount of monoterpenes of the
myrtle essential oils from leaves collected in Italy and Turkey was similar but the qualitative
 Felice Senatore et al. / Jeobp 9 (2) 2006 pp 162 - 169 165

composition was different. In fact, the compounds were oxygen containing monoterpenes
(alcohols, acetates and one oxide), monoterpene hydrocarbons and a small amounts of ses-
quiterpenes and phenols. Both oils were characterized by high contents of monoterpenes,
but in the oil from Italy monoterpene hydrocarbons (50.8%) were the most representative
components and α-pinene (15.7%) was the main compound of this fraction. 1,8-Cineole
(16.5%) and linalool (8.9%) constituted about the two thirds of the oxygen containing monot-
erpenes (38.5%).
On the contrary, the oil from Turkey showed low content of monoterpene hydrocar-
bons (9.0%) with α-pinene (7.8%) as the main compound being the others components of
this fraction present in low concentration or traces, and was characterized by an higher
content of oxygen containing monoterpenes (81.6%) with linalool (36.5%) and linalyl ac-
etate (16.3%) as the most representative compounds. Other compounds of this fraction
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present in interesting amounts were 1,8-cineole (10.5%), geranyl acetate (8.0%) and α-
terpineol (7.9%). Differences were also found for the composition of the sesquiterpenoidic
fractions: though they were present in the same concentration, their composition is different.
The oil from Turkey was characterized by the absence of oxygenated sesquiterpenes and
contains six sesquiterpene hydrocarbons in quite low (2.8%) content, ranging between 0.9%
(caryophyllene) and trace amounts (δ-elemene). The oil from Italy showed the same con-
tent (2.9%) of sesquiterpene hydrocarbons with caryophyllene (2.0%) as the most important
component. Spathulenol (0.1%) was the unique oxygen containing sesquiterpene identified
in this oil. The composition of the present oils differs from that reported for the oils obtained
from leaves and branches of myrtle collected in Italy (Liguria24 and Sardinia26) and in Tur-
key11 and this finding agrees with the hypothesis that the composition of the essential oils is
also function of the harvest time and environmental factors27,35,36.
The results obtained in the antimicrobial assay are presented in Table 2. The oils from
Italy and Turkey showed both a good antimicrobial activity, but the Turkish oil was slightly
more active than the Italian one, especially against Gram + bacteria. This finding can be
justified by a major concentration in it of active compounds such as linalool, α-terpineol,
geranyl acetate. The oils had a good activity (12,5 µg/mL) against Staphylococcus
epidermidis. Among Gram + bacteria, Staphylococcus aureus appeared to be the less
sensitive (100 µg/mL). The oils showed a similar activity against Bacillus cereus and Strep-
tococcus faecalis, with MICs of 50 and 25 µg/mL respectively. Among Gram – bacteria,
Escherichia coli was the most sensitive (25 µg/mL). A slight activity of both the oils was
detected against Proteus vulgaris and Salmonella typhi Ty2 (100 µg/mL), while there was
no activity at all against Pseudomonas aeruginosa (>100 µg/mL). The antimicrobial nature
of myrtle essential oils can be attribuited to the presence of various substances, mainly 1,8-
cineole, α-pinene, linalyl acetate, linalool, α-terpineol, geranyl acetate, but it is also attribut-
able to a synergistic antimicrobial effect of these compounds34,35. The good activity of myrtle
essential oil against different bacterial strain may contribute to the medicinal properties of
M. communis.

Acknowledgment: The GC and GC-MS spectra were performed at the C.S.I.A.S.

of the University “Federico II” of Naples. The assistance of the staff is gratefully appreci-
ated.
 Felice Senatore et al. / Jeobp 9 (2) 2006 pp 162 - 169 166

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 Felice Senatore et al. / Jeobp 9 (2) 2006 pp 162 - 169 168

Table 1 - Chemical composition of essential oil of Myrtus communis L. growing

wild in Italy (I) and Turkey (T).

K ia Component Identificationb I %c T% c

914 Tricyclene Ri, Co-GC, MS - t

929 α-Thujene Ri, MS 0.3 t
938 α-Pinene Ri, Co-GC, MS 15.7 7.8
954 Camphene Ri, MS 0.2 -
974 Sabinene Ri, Co-GC, MS 0.6 t
978 β-Pinene Ri, Co-GC, MS 1.6 t
988 Myrcene Ri, Co-GC, MS 0.4 0.7
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1005 α-Phellandrene Ri, MS 1.5 -

1013 ∆ 3 -Carene Ri, MS 1.5 t
1018 γ-Terpinene Ri, MS, Co-GC 2.1 -
1026 p-Cymene Ri, MS, Co-GC 3.3 -
1027 trans-Pinocarveol Ri, MS - 0.3
1030 Limonene Ri, MS, Co-GC 8.9 -
1033 1,8-Cineole Ri, MS, Co-GC 16.5 10.5
1038 (Z)-β-Ocimene Ri, MS - 0.4
1049 (E)-β-Ocimene Ri, MS 1.0 -
1059 γ-Terpinene Ri, MS, Co-GC 3.5 0.1
1089 Linalool Ri, MS, Co-GC 8.9 36.5
1090 Terpinolene Ri, MS 0.2 -
1148 Camphor Ri, MS, Co-GC t -
1168 Borneol Ri, MS, Co-GC 2.2 -
1178 Terpineol-4 Ri, MS, Co-GC 0.7 t
1187 α-Terpineol Ri, MS, Co-GC 2.7 7.9
1190 Myrtenal Ri, MS 1.1 -
1194 Myrtenol Ri, MS 0.6 1.0
1248 Linalyl acetate Ri, MS, Co-GC 13.2 16.3
1264 Bornyl acetate Ri, MS 0.2 t
1278 Carvacrol Ri, MS, Co-GC - 0.8
1333 α-Terpinyl acetate Ri, MS 0.6 1.1
1337 δ-Elemene Ri, MS 0.1 t
1367 Methyl eugenol Ri, MS 2.5 1.7
1368 Geranyl acetate Ri, MS 1.8 8.0
1419 Caryophyllene Ri, MS, Co-GC 2.0 0.9
1454 α-Humulene Ri, MS 0.3 0.7
1460 allo-Aromadendrene Ri, MS 0.5 -
1468 γ-Muurolene Ri, MS t 0.2
1485 Germacrene D Ri, MS - 0.4
1499 (Z)-α-Bisabolene Ri, MS - 0.6
 Felice Senatore et al. / Jeobp 9 (2) 2006 pp 162 - 169 169

table 1. (continued0

K ia Component Identificationb I %c T% c

1575 Spathulenol Ri, MS 0.1 -

Monoterpene hydrocarbons 40.8 9.0

Oxygen containing monoterpenes 48.5 81.6
Sesquiterpene hydrocarbons 2.9 2.8
Phenols 2.5 2.5
Total identified 94.8 95.9
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a: Retention index on a DB-5 column;

b: Ri = retention index,
MS = mass spectrum,
Co-GC = co-injection with authentic compound;
c: T = trace, less than 0.05%.

µg/mL) of essential oils from Myrtus communis L. aerial

Table 2. MIC values (µ
parts and MIC of reference antibiotic.

Bacterial strain I T G*

Bacillus subtilis 50 25 1.56

ATCC 6633
Staphylococcus aureus 100 100 3.12
ATCC 25923
Staphylococcus epidermidis 12.5 12.5 6.25
ATCC 12228
Streptococcus faecalis 50 25 >100
ATTC 29212
Escherichia coli 25 25 3.12
ATCC 25922
Proteus vulgaris 100 100 100
ATCC 13315
Pseudomonas aeruginosa >100 >100 12.5
ATCC 27853
Salmonella typhi Ty2 100 100 >100
ATCC 19430

*: G = gentamicyne