The composition and antibacterial activity of the essential oil of Levisticum officinale Koch flowers and fruits at different developmental stages

Abstract

The composition and antibacterial activity of the essential oil of Levisticum officinale Koch at different developmental stages (flower, immature fruit, green mature fruit and ripened fruit) is reported. The essential oils were obtained by hydrodistillation of air-dried samples and their antibacterial activities were tested against seven bacteria. The yield of oil (w/w %) in different stages was in the order: immature fruit (1.5 %) > green mature fruit (1.0 %) > ripened fruit (0.6 %) > flower (0.1 %). The essential oils were analyzed by GC and GC–MS. In total, 27, 31, 28 and 26 constituents were identified and quantified in the mentioned samples, respectively. Monoterpene hydrocarbons were the main group of compounds in the green mature fruit (79.2 %), immature fruit (78.4 %), ripened fruit (75.2 %) and flower (44.0 %). The antibacterial activity of the oils was evaluated by the disk diffusion method using Müller–Hinton agar and determination of inhibition zones. The results of the bioassays showed some variations between the three tested oils in their inhibitory activity against the tested bacteria at a 10 µl disc–1 concentration. The oils from mature and ripened fruit exhibited potent antibacterial activity against Bacillus subtilis, with minimum inhibitory concentration (MIC) values of 0.90 mg ml–1 in mature and ripened fruits.

Full text

J. Serb. Chem. Soc. 75 (12) 1661–1669 (2010) UDC Levisticum officinale:665.52:615.281–188
JSCS–4086 Original scientific paper
1661
The composition and antibacterial activity of the essential oil
of Levisticum officinale Koch flowers and fruits at different
developmental stages
MOHAMMAD HOSSEIN MIRJALILI1*, PEYMAN SALEHI1, ALI SONBOLI1,
JAVAD HADIAN1, SAMAD NEJAD EBRAHIMI1 and MORTEZA YOUSEFZADI2
1Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, G. C.,
Evin, Tehran and 2Department of Marine Biology, Faculty of Science, Hormozgan
University, Bandar Abbas, Iran
(Received 24 May, revised 9 July 2010)
Abstract: The composition and antibacterial activity of the essential oil of Le-
visticum officinale Koch at different developmental stages (flower, immature
fruit, green mature fruit and ripened fruit) is reported. The essential oils were
obtained by hydrodistillation of air-dried samples and their antibacterial acti-
vities were tested against seven bacteria. The yield of oil (w/w %) in different
stages was in the order: immature fruit (1.5 %) > green mature fruit (1.0 %) >
> ripened fruit (0.6 %) > flower (0.1 %). The essential oils were analyzed by
GC and GC–MS. In total, 27, 31, 28 and 26 constituents were identified and
quantified in the mentioned samples, respectively. Monoterpene hydrocarbons
were the main group of compounds in the green mature fruit (79.2 %), im-
mature fruit (78.4 %), ripened fruit (75.2 %) and flower (44.0 %). The anti-
bacterial activity of the oils was evaluated by the disk diffusion method using
Müller–Hinton agar and determination of inhibition zones. The results of the
bioassays showed some variations between the three tested oils in their inhi-
bitory activity against the tested bacteria at a 10 µl disc–1 concentration. The
oils from mature and ripened fruit exhibited potent antibacterial activity against
Bacillus subtilis, with minimum inhibitory concentration (MIC) values of 0.90
mg ml–1 in mature and ripened fruits.
Keywords: Levisticum officinale Koch; Apiaceae; essential oil; antibacterial
activity; reproductive stage.
INTRODUCTION
Lovage (Levisticum officinale Koch) is a perennial herbaceous plant from
the Apiaceae family with origins in Iran and Afghanistan; it can now be found
throughout the world.1–4 The plant has been alternatively classified as Ligus-
* Corresponding author. E-mail: m-mirjalili@sbu.ac.ir
doi: 10.2298/JSC100524126M
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1662 MIRJALILI et al.
ticum levisticum L., Hipposelinum levisticum Britt. and Angelica levisticum Bail-
lon.5 The essential oil of roots, seeds and leaves of lovage are used in a wide va-
riety of applications including food flavoring, medicinal preparations, aromathe-
rapy, perfumery and industrial fragrances.2,6,7 Moreover, the plant is used in
Iranian folk medicine for the treatment of several gastrointestinal, nervous and
rheumatic disorders.2,8 The essential oil composition of the plant was previously
studied in different countries and more than 190 compounds were reported in its
root, seed or leaf oil.9 It was found that the chemical compositions of the es-
sential oils distilled from separate botanical parts of this plant are rather diffe-
rent.10–12 The chemical constituents of lovage root oil are mainly phthalides in-
cluding n-butylidene phthalide and n-butyl-phthalide, sedanonic anhydride, ter-
penoids such as α-terpineol, carvacrol, phenylpropanoids such as eugenol and
volatile acids.5,13,14 Polyacetylenes as antimycobacterial compounds have also
been reported from the plant.15 The effect of harvesting time, plant age, cutting
frequency and the method of plantation establishment on the essential oil yield
and components in different parts of L. officinale was investigated previous-
ly.7,11,16,17 It was found that the flowers and seeds produced the highest yields of
the oil with β-phellandrene (40.8 and 61.5 %, respectively) as the main con-
stituent, while α-terpinyl acetate (≈70.0 %) was reported as the principal con-
stituent of the leaves and stems oils.7 In another study, the oil of lovage fruits
contained β-phellandrene (69.3 %),
α
-terpinenyl acetate (4.2 %) and α-terpineol
(2.1 %) as the major components.12 It was reported that the essential oil content
was similar in roots, stems, petioles, leaves and inflorescence, while the highest
content was found in seeds (1.9 %).18 Seasonal variations in the composition of
headspace volatiles were also determined,10 of which, β-phellandrene was the
most abundant component in all plant parts except for root. Samiee et al. reported
terpinyl acetate (40.5 %) and β-phellandrene (16.7 %) as the main constituents in
the essential oil and β-phellandrene (23.0 %), naphthalene (20.6 %) and γ-ter-
pinene (12.1 %) as the major components in the methanol extract of the plant
from Iran.19 Recently, (Z)-falcarinol, n-octanal, palmitic acid, (Z)-ligustilide, (Z)-
-3-butylidenephthalide, trans-β-farnesene have been reported as the main com-
pounds of the essential oil of hairy root cultures of L. officinale.20–22 Variations
in the essential oil composition of roots and leaves of L. officinale from different
European countries have also been studied. Ten compounds, including trans-p-
-mentha-2,8-dien-1-ol, iso-thujyl alcohol, p-mentha-1,5-dien-8-ol, bicyclo[3.2.0]-
heptan-3-ol, 2-methylene-6,6-dimethyl, trans-carveol, perillaldehyde, sabinyl ace-
tate, perillyl alcohol, the methyl ester of methylpentadecanoic acid and methyl-
hexadecadienoic acid, were introduced for the first time.23 To the best of our
knowledge, there is no previous report on the essential oil analysis and antibac-
terial activity of L. officinale at different developmental stages. Thus, in this pa-
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ESSENTIAL OIL O F Levisticum officinale Koch 1663
per, the composition and antibacterial activity of the essential oils of this plant at
different stages of its development are reported.
EXPERIMENTAL
Plant material
These experiments were conducted during 2007–2009 at the field of the Medicinal Plants
and Drugs Research Institute of Shahid Beheshti University, located in Evin (35°48’ N,
51°23’ E at an altitude of 1785 m) in the north of Tehran, Iran. The plant seeds were obtained
from the seed bank of the Medicinal Plants and Natural Products Research Institute, Iranian
Academic Center for Education, Culture and Research (ACECR) and were sown in a
greenhouse in the last week of February, 2007. Nine-week-old seedlings were transplanted at
50 cm row-to-row and 30 cm plant-to-plant spacing in the experimental field in May, 2007.
The sampling was realized from a 2-year-old cultivated population by the random collection
of 10 individuals for each developmental stage. For the collection of the flowers, all of them
on the inflorescence were opened. The samples at the fruiting stage were collected at three
different times of fruit maturation, i.e. immature (infructescence with young fruits 15 days
after flowering), mature (infructescence with solid and dark green colored fruit) and ripened
(infructescence with yellowish fruits just in the deciduous time). Voucher specimens (No.
200364-7) representative of each sample were deposited at the Medicinal Plants and Drugs
Research Institute Herbarium (MPH), Shahid Beheshti University of Tehran.
Essential oil isolation procedure
The essential oil of air-dried samples (100 g) of each stage was isolated by hydrodistil-
lation for 3 h, using a Clevenger-type apparatus according to the method recommended in
British Pharmacopoeia (1993).24 The isolated oils were dried over anhydrous sodium sulfate
and stored in dark tightly closed vials at 4 °C until analysis.
Essential oil analysis procedure
GC analysis was conducted using a Varian CP-3800 instrument equipped with a DB-1
fused silica capillary column (25 m×0.25 mm i.d., film thickness 0.25 μm). Nitrogen was used
as the carrier gas at a constant flow rate of 1.1 ml min–1. The oven temperature was held at 60
°C for 1 min, then programmed to 250 °C at a rate of 4 °C min-1, and then held for 10 min.
The injector and detector (FID) temperatures were kept at 250 and 280 °C, respectively. GC/
/MS analysis was realized on a Thermoquest-Finnigan Trace GC/MS instrument equipped
with a DB-1 fused silica column (60 m×0.25 mm i.d., film thickness 0.25 μm). The oven tem-
perature was raised from 60 to 250 °C at a rate of 5 °C min-1 and then held at 250 °C for 10
min; the transfer line temperature was 250 °C. Helium was used as the carrier gas at a flow
rate of 1.1 ml min-1; the split ratio was 1/50. The quadrupole mass spectrometer was scanned
over the 45–465 amu range with an ionizing voltage of 70 eV and an ionization current of 150 μA.
Identification and quantification of the oil components
The constituents of the essential oils were identified by calculation of their retention
indices under temperature-programmed conditions for n-alkanes (C6–C24) and the oil on a
DB-1 column under the same chromatographic condition. Identification of individual com-
pounds was made by comparison of their mass spectra with those of the internal reference
mass spectra library or with authentic compounds and confirmed by comparison of their
retention indices with authentic compounds (purchased from Sigma-Aldrich and Merck) or
with those reported in the literature.25 For quantification purpose, the relative area percentages
obtained by FID were used without the use of correction factors.
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1664 MIRJALILI et al.
Antibacterial activity
The antibacterial activity of the oils was evaluated by the disk diffusion method using
Müller-Hinton agar26 and determination of the inhibition zones. The essential oils were tested
at a concentration of 10 μl per disk. The microorganisms used were as follows: Bacillus sub-
tilis ATCC 9372, Enterococcus faecalis ATCC 15753, Staphylococcus aureus ATCC 25923,
Staphylococcus epidermidis ATCC 12228, Escherichia coli ATCC 25922, Pseudomonas ae-
ruginosa ATCC 27852, and Klebsiella pneumoniae ATCC 3583. For the determination of the
minimum inhibitory concentration (MIC), a microdilution broth susceptibility assay was used,
as recommended by NCCLS.27 The technical data were described previously.28 Ampicillin
was used as the standard reference for the positive control.
RESULTS AND DISCUSSION
Essential oil analysis
The essential oils had a light yellow color with distinct sharp odor. The yield
of the essential oils (w/w %) of the plant at different developmental stages were
in the order: immature fruit (1.5 %) > green mature fruit (1.0 %) > ripened fruit
(0.6 %) > flower (0.1 %). The qualitative and quantitative analytical results are
listed in Table I together with the retention indices of the identified compounds,
where all the constituents are arranged in order of their elution on the DB-1 co-
lumn. In total 27, 31, 28 and 26 constituents, respectively, were identified and
quantified in the studied samples representing 95.9, 99.9, 98.7 and 92.7 % of the
total oil, respectively. A comparison among the composition of the essential oils
revealed both quantitative and qualitative differences. The GC and GC–MS ana-
lyses showed that the distribution of saturated hydrocarbons of the oil from flo-
wer was remarkably different from that of the oils at the fruiting stage. The re-
sults revealed that the saturated hydrocarbons from the flower (12.3 %) were
present in higher amount than in the other samples. Heneicosane (6.0 %) and tri-
cosane (3.0 %) were found only in the oil of the flower. The major constituent of
the oil of the flower was β-pinene (17.7 %), but it was found that this compound
decreased gradually in subsequent developmental stages. The β-pinene and α-pi-
nene contents were highest in the essential oil of the first harvest and decreased
with progressive maturation of the fruit. On the contrary, β-phellandrene was
found as the principal component of the oil after fruit initiation, i.e., it constituted
11.7 % of the oil of the flower but increased remarkably in the fruit oils, con-
stituting 62.4, 60.5 and 56.4 % of the green mature, immature and ripened fruit
oils, respectively. β-Phellandrene has already been reported as the main consti-
tuent in the essential oil from the flowers and fruits in previous reports.7,10,12
β-Gurjunene (2.8 %), globulol (0.7 %) and geranyl acetate (3.3 %) were found
only in the oil of the flower. α-Phellandrene, δ-elemene and germacrene-D were
absent completely in the oil of flower but were present in trace or low amounts in
the other samples. The essential oil obtained from immature fruit contained the
highest contents of sabinene (2.3 %), isomenthol (5.6 %), cis-dihydrocarvon (0.6
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ESSENTIAL OIL O F Levisticum officinale Koch 1665
%), germacrene-D (0.6 %), elemol (0.5 %) and trans-nerolidol (1.6 %) compared
with the other samples.
TABLE I. Composition of the essential oil of Levisticum officinale at different developmental
stages
RI
a Compound From flowers
% In fruiting stages, % Identification
methods
Immature Mature Ripened
0935 α-Pinene 5.3 4.6 4.3 2.9 RI, MS
b
, CoIc
0969 Sabinene 1.7 2.3 1.5 1.1 RI, MS
0976 β-Pinene 17.7 11.5 4.1 2.9 RI, MS, CoI
0982 Myrcene 1.3 t
d
t t RI, MS
1002 α-Phellandrene – t t t RI, MS
1010 δ-3-Carene 3.0 2.8 5.7 6.8 RI, MS
1017 ortho-Cymene 1.4 t t t RI, MS
1026 β-Phellandrene 11.7 56.4 62.4 60.5 RI, MS
1038 cis-Ocimene 1.9 0.8 0.8 0.8 RI, MS
1083 Terpinolene – – 0.4 0.2 RI, MS, CoI
1111 cis-p-Menth-2-en-1-ol – 0.3 – 0.2 RI, MS
1162 Isomenthol 1.8 5.6 3.9 2.1 RI, MS
1166 4-Terpineol 0.5 0.6 – – RI, MS
1168 cis-Dihydrocarvone – 0.6 – – RI, MS
1217 Cuminyl aldehyde – 0.5 t – RI, MS
1265 p-Cymene-7-ol – 0.4 t – RI, MS
1339 δ-Elemene – 0.4 0.3 0.3 RI, MS
1358 Geranyl acetate 3.3 – – – RI, MS, CoI
1385 α-Copaene 1.3 0.3 0.3 0.2 RI, MS
1392 β-Elemene 1.0 1.2 1.3 1.3 RI, MS
1433 α-Humulene 0.9 0.4 0.3 0.2 RI, MS
1436 β-Gurjunene 2.8 – – – RI, MS
1446 (Z)-β-Farnesene 4.3 0.2 0.2 0.2 RI, MS
1473 γ-Curcumene 8.0 0.2 0.2 0.3 RI, MS
1484 Germacrene-D – 0.6 0.3 t RI, MS
1489 Zingiberene 0.8 0.5 0.8 t RI, MS
1500 Germacrene-B 0.8 1.6 t 1.2 RI, MS
1503 β-Bisabolene 1.6 0.8 1.8 – RI, MS
1518 β-Sesquiphellandrene 2.1 1.2 2.4 5.1 RI, MS
1541 Elemol – 0.5 – – RI, MS
1548 trans-Nerolidol 0.8 1.6 – – RI, MS
1562 γ-Elemene – 0.8 0.8 1.2 RI, MS
1574 Spathulenol 8.9 1.1 1.9 2.7 RI, MS
1593 Globulol 0.7 – – – RI, MS
1601 Hexadecane 3.3 2.1 2.5 2.2 RI, MS
1691 3-iso-Thujopsanone – – 2.5 0.3 RI, MS
2097 Heneicosane 6.0 – – – RI, MS
2301 Tricosane 3.0 – – – RI, MS
–
Monoterpene
hydrocarbons 44.0 78.4 79.2 75.3 –
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1666 MIRJALILI et al.
TABLE I. Continued
RI
a Compound From flowers
% In fruiting stages, % Identification
methods
Immature Mature Ripened
–
Oxygenated
monoterpenes 8.9 8.0 3.5 2.3
–
Sesquiterpene
hydrocarbons 20.3 9.0 9.1 10.7
–
Oxygenated
sesquiterpenes 10.4 2.4 4.4 2.2
–
Other 12.3 2.1 2.5 2.2
Total identified 95.9 99.9 98.7 92.7
aRetention indices relative to C6–C24 n-alkanes on a DB-1 column; bmass spectroscopy; cco-injection with au-
thentic compounds; dtrace, less than 0.1 %
The classification of the identified compounds based on functional groups is
summarized at the end of Table I and shows that monoterpene hydrocarbons
were the main group of compounds in all samples. The monoterpene hydrocar-
bons content was the lowest in the flower and increased with subsequent harvest-
ing times to reach maximum in the mature fruit and then decreased in the ripened
fruit. In this study, the oil from green mature fruit contained β-phellandrene in
higher amount (62.4 %) than the oils from ripened fruit (60.5 %), immature fruit
(56.4 %) and flower (11.7 %). The other major monoterpene hydrocarbons which
were found in all samples were β-pinene, α-pinene and δ-3-carene, while in ano-
ther study,
α
-terpinenyl acetate, α-terpineol, limonene and myrcene were reported
as the major monoterpenes.12 Monoterpene hydrocarbons identified in the oil of
flower were present in lower amount than in the oil of other stages. On the other
hand, in the essential oil of flower, sesquiterpenes were one of the dominant frac-
tion with spathulenol (8.9 %) as the major compound and their percentage de-
creased with progressive fruit maturation.
Antibacterial activity
The disk diffusion method, used in preliminary screening of the antibacterial
activity, showed that the oils from the three different fruiting stages of L. offici-
nale were active against all the tested bacteria. Moreover, the oils proved to be
highly active against the tested Gram-positive bacteria, especially B. subtilis that
was more sensitive than others, and the Gram-negative bacterium, E. coli (Table
II). The oils were moderately active against K. pneumoniae and P. aeruginosa.
The antibacterial activities of the oils were also determined using the microtiter
96-well dilution method, by measuring the minimal inhibitory concentration (MIC)
against the tested bacteria (Table II). The essential oils of mature and ripened
fruits exhibited the highest activity against B. subtilis with an MIC value of 0.90
mg ml–1. In addition, the highest activity of the oil of mature fruit was observed
against S. epidermidis, with an MIC value of 0.90 mg ml–1. The oils showed the
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ESSENTIAL OIL O F Levisticum officinale Koch 1667
lowest activity against K. pneumoniae and P. aeruginosa, with the MIC values
ranging from 14.4 to 15.2 mg ml–1.
TABLE II. Antibacterial activity of L. officinale essential oil at different fruiting stages
Microorganism Essential oila Ampicillinb
Immature Mature Ripened
DDc MIC
d
DD MIC DD MIC DD
B
. subtilis 25.0±0.9 3.8 36.0±0.5 0.9 35.0±0.4 0.9 14.0±0.7
E
. faecalis 19.0±0.7 15.2 17.0±0.4 7.5 13.0±0.4 7.2 11.0±0.4
S. aureus 17.0±0.5 3.8 21.0±0.5 3.7 16.0±0.5 3.6 13.0±0.6
S. epidermidis 23.0±0.7 1.9 26.0±0.5 0.9 25.0±0.6 1.8 19.0±0.5
E
. coli 19.0±0.6 15.2 18.0±0.4 7.2 15.0±0.7 7.2 12.0±0.5
K
. pneumoniae 10.0±0.8 >15.2 10.0±0.7 >14.4 9.0±0.4 >14.4 –
P
. aeruginosa 11.0±0.7 >15.2 8.0±0.6 >14.4 9.0±0.5 >14.4 9.7±0.7
aTested at a concentration of 10 μl disc-1; btested at a concentration of 10 μg disc-1; cdiameter of inhibition zone
(mm) including disk diameter of 6 mm: inactive (–), moderately active (7–14) and highly active (>14); dmini-
mum inhibitory concentration, values as mg ml oil-1
CONCLUSIONS
The study of a plant as a source of aromatic and flavoring compounds re-
quires the analysis of not only the whole plant but also its individual parts at their
different developmental stages. While the best harvesting time of lovage to ob-
tain sharp odorant compounds such as α- and β-pinene is the flowering stage,
β-phellandrene as the main compound of the plant with a peppery-minty and
slightly citrusy odor was achieved at the fruiting stage. Chemical characterization
and antibacterial screening studies on the plant-based essential oils could also
lead to the discovery of new natural antibacterial agents. In addition to perfume
and tobacco products, the essential oil of lovage is used as a flavor agent in major
food products, such as beverages, frozen dairy desserts, candy, gelatins and pud-
ding, and meat and its products. Although the antimycobacterial activity of poly-
acetylenes, such as falcarinol and falcarindiol, from the plant has recently been
studied,15 the present study is the first report on the antibacterial activity of the
essential oil from fruits of L. officinale at different developmental stages. The oils
showed promising antibacterial activity against, B. subtilis and S. epidermidis.
The present results revealed that the essential oils tested represent an inexpensive
source of natural antibacterial substances for use in pathogenic systems to pre-
vent the growth of bacteria and extend the shelf life of processed food.
Acknowledgement. We are grateful to Shahid Beheshti University Research Council for
financial support of this work.
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1668 MIRJALILI et al.
ИЗВОД
САСТАВ И АНТИБАКТЕРИЈСКА АКТИВНОСТ ЕТАРСКОГ УЉА ЦВЕТА И ПЛОДА
БИЉКЕ Levisticum officinale Koch У РАЗЛИЧИТИМ РАЗВОЈНИМ ФАЗАМА
MOHAMMAD HOSSEIN MIRJALILI1, PEYMAN SALEHI1, ALI SONBOLI1, JAVAD HADIAN1,
SAMAD NEJAD EBRAHIMI1 и MORTEZA YOUSEFZADI2
1Medicinal Plants and Drugs Research Institute, Shahid Beheshti University,G. C., Evin, Tehran и 2Department
of Marine Biology, Faculty of Science, Hormozgan University, Bandar Abbas, Iran
Одређиван је састав и антибактеријска активност етарског уља биљке Levisticum offici-
nale Koch у различитим развојним фазама: цвет, незрео плод, зелени зрео плод и потпуно
зрео плод. Етарско уље је добијено из сувих узорака дестилацијом воденом паром, а анти-
бактеријска активност је одређивана спрам седам врста бактерија. Принос уља (масени %) у
различитим фазама је био следећи: незрео плод (1,5 %) > зелени зрео плод (1,0 %) > потпуно
зрео плод (0,6 %) > цвет (0,1 %). Састав етарског уља је одређиван методама GC и GC–MS. У
ова четири узорка је идентификовано и квантификовано редом 27, 31, 28 и 26 састојака. Мо-
нотерпенски угљоводоници су чинили главну групу једињења: 79,2 % у зеленом зрелом пло-
ду, 78,4 % у незрелом плоду, 75,2 % у потпуно зрелом плоду и 44,0 % у цвету. Антибакте-
ријска активност уља је одређивана методом прстенасте дифузије у Милер-Хинтоновом ага-
ру мерећи зону инфибиције. Коришћено је 10×10 µl уља за инхибицију и резултати су били
различити за уља добијена из биљке у различитим развојним фазама. Најјача антибакте-
ријска активност је испољена спрам Bacillus subtilis. MIC вредност је била 0,90 mg ml-1 са
уљем потпуно зрелог плода.
(Примљено 24. маја, ревидирано 9. јула 2010)
REFERENCES
1. K. H. Rechinger, in Flora Iranica, K. H. Rechinger, Ed., Akademische Drucku Verlags-
anstalt, Graz, 1987
2. M. H. Mirjalili, J. Javanmardi, in Handbook of Herbs and Spices, K. V. Peter, Ed.,
Woodhead Publishing Ltd., Abington, 2006
3. V. Mozaffarian, A Dictionary of Iranian plant names, Farhang Moaser, Tehran, 1996
4. E. Daukšas, R. P. Venskutonis, B. Sivik, J. Supercrit. Fluids 22 (2002) 201
5. J. E. Simon, A. F. Chadwick, L. E. Craker, The Scientific Literature on Selected Herbs,
Aromatic and Medicinal Plants of the Temperate Zone, Archon Books, Hamden, CT, 1984
6. B. Toulemonde, I. Noleau, Dev. Food Sci. 18 (1988) 641
7. E. Bylaite, R. P. Venskutonis, J. P. Roozen, J. Agric. Food Chem. 46 (1998) 3735
8. A. Zargari, Medicinal plants, Tehran University Publications, Tehran, 1990
9. E. Daukšas, R. P. Venskutonis, B. Sivik, T. Nilson, J. Supercrit. Fluids 15 (1999) 51
10. E. Bylaite, J. P. Roozen, A. Legger, R. P. Venskutonis, M. A. Posthumus, J. Agric. Food
Chem. 48 (2000) 6183
11. I. Novak, E. Nemeth, Acta Hort. 576 (2002) 311
12. J. Dyduch, A. Najda, T. Wolski, S. Kwiatkowski, Folia Hort. 15 (2003) 141
13. M. J. M. Gijbels, J. J. C. Scheffer, A. B. Svendsen, Planta Med. 44 (1982) 207
14. M. E. Ibrahim, Egypt. J. Hort. 26 (1999) 177
15. A. Schinkovitz, M. Stavri, S. Gibson, F. Bucar, Phytother. Res. 22 (2008) 681
16. C. Rohricht, Z. Arznei-Gewurzpflanzen 14 (2009) 105
17. S. Andruszczak, Acta Sci. Pol.-Hortoru. 6 (2007) 21
2010 Copyright (CC) SCS
_________________________________________________________________________________________________________________________
Available online at www.shd.org.rs/JSCS/

ESSENTIAL OIL O F Levisticum officinale Koch 1669
18. K. Seidler-Lozykowska, K. Kazmierczak, Herba Polonica 44 (1998) 11
19. K. Samiee, M. R. Akhgar, A. Rustaiyan, S. Masoudi J. Essent. Oil Res. 18 (2006) 19
20. S. Nunes, J. M. S. Faria, A. C. Figueiredo, L. G. Pedro, H. Trindade, J. G. Barroso Planta
Med. 75 (2009) 387
21. M. Costa, A. C. Figueiredo, J. G. Barroso, L. G. Pedro, S. G. Deans, J. J. C. Scheffer
Biotechol. Lett. 30 (2008) 1265
22. P. A. G. Santos, A. C. Figueiredo, M. M. Oliviera, J. G. Baroso, L. G. Pedro, S. G. Deans,
J. J. C. Scheffer, Plant Sci. 168 (2005) 1089
23. A. Raal, E. Arak, A. Orav, T. Kailas, M. Müürisepp, J. Essent. Oil Res. 20 (2008) 318
24. British pharmacopoeia, HMSO, London, 1993
25. T. Shibamoto, in Capillary Gas Chromatography in Essential Oil Analysis, P. Sandra, C.
Bicchi. Eds., Hüthig Verlag, New York, 1987, p. 259
26. E. J. Baron, S. M. Finegold, Bailey and Scott’s Diagnostic Microbiology, 8th ed., C.V.
Mosby Co., St. Louis, MO, 1990
27. National Committee for Clinical Laboratory Standards, Performance standards for anti-
microbial susceptibility testing, 9th international supplement, M100-S9, National Com-
mittee for Clinical Laboratory Standards, Wayne, PA, 1999
28. D. Azaz, F. Demirci, F. Satil, M. Kurkcuoglu, K. H. C. Baser, Z. Naturforsch. 57c (2002)
817.
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