Chemical composition, antimicrobial activities and
olfactory evaluations of an essential marjoram oil from
Albania as well as some target compounds
Chemische Zusammensetzung, antimikrobielle Aktivitäten und
olfaktorische Beurteilungen eines ätherischen Majoranöls aus
Albanien sowie einiger Schlüsselkomponenten
L. Jirovetz, S. Bail, G. Buchbauer, Z. Denkova, A. Slavchev, A. Stoyanova, E. Schmidt
The chemical composition of an essential marjoram oil from
Die chemische Zusammensetzung eines kommerziell herge-
Albania produced for commercial purposes was analysed
stellten ätherischen Majoranöls aus Albanien wurde mittels
by GC and GC/MS. 28 volatiles were identiﬁed and the fol-
GC und GC/MS analysiert. 28 ﬂüchtige Inhaltsstoffe wurden
lowing monoterpenes were found as main compounds: ter-
identiﬁziert und als Hauptkomponenten die Monoterpe-
pinen-4-ol (21.33%), trans
-sabinene hydrate (15.53%), γ-ter-
ne Terpinen-4-ol (21,33 %), trans
-Sabinenhydrat (15,53 %),
pinene (14.00%), α-terpinene (8.88%) and sabinene (8.27%).
γ-Terpinen (14,00 %), α-Terpinen (8,88 %) und Sabinen
The essential Origanum majorana
oil, some higher and
(8,27 %) gefunden. Die antimikrobielle Aktivität des äthe-
medium-concentrated constituents, a phenolic reference
Öls sowie einiger Haupt- und
component and three antibiotics were tested against sev-
Nebenkomponenten wurden mit einem optimierten und
eral microorganisms using modiﬁed and well-established
bewährten Agardiffusions- und Agarverdünnungstest be-
and optimised agar dilution and agar diffusion assays to
stimmt. Zum direkten Vergleich wurden drei Antibiotika so-
evaluate the antimicrobial activity of the oil as well as these
wie eine phenolische Referenzkomponente hinzugezogen.
target compounds. In addition, the analytical results were
Die analytischen Resultate wurde mit olfaktorischen Bewer-
correlated with olfactoric evaluations for reasons of quality
tungen zur Qualitätskontrolle dieses ätherischen Majoran-
control of the essential marjoram oil.
Essential oil, Origanum majorana
L., chemical composition,
Ätherisches Öl, Origanum majorana
L., chemische Zusam-
antimicrobial activities, olfactoric evaluations
mensetzung, antimikrobielle Aktivitäten, olfaktorische Beur-
vouring of foodproducts, the applications of these oils in perfumery and cosmetics as well as the biological,
In continuation of an international project in the ﬁeld of
pharmacological and toxicological effects as fresh
combined data interpretation of composition analysis,
plant, herb, tea, extract and especially essential oil for
odour evaluation and antimicrobial activity testings of
the treatment of various diseases are available so far
various aroma samples , an essential oil of marjo-
ram (Origanum majorana
L., syn. Majorana hortensis
– common names: English: mar-
This immense number of chemical and antimicrobial data
joram, sweet marjoram, knotted marjoram; Arabic:
is based on different non-commercial marjoram samples.
bardagoush, barsagusha, mardqouche, mizunjuske;
The aim of this respective study was to analyze an es-
Dutch: marjolein; French: marjolaine, origane; German:
sential oil of Origanum majorana
from Albania produced
Gartendost, echter Majoran, Meiran, Wurstkraut [2, 3]
under commercial manufacturing conditions by means
– from Albania was investigated. Many papers about
of GC and GC/MS analysis, olfactory evaluation and anti-
the composition of Origanum majorana
, its extracts
microbial testings, using well-established and optimised
and essential oils, its utilisation as spice and for ﬂa-
agar diffusion and agar dilution methods .
2 Materials and methods
For other parameters, see description of GC/FID, above. Mass spectra correlations were done using
2.1 Samples and reference compounds
Wiley, NBS, NIST and our own library as well as pub-
Sabinene hydrate (purity: 98.1%, product no.: 96573)
is a product of Fluka-Riedel-de Haen-Sigma Aldrich
(Buchs, Switzerland); α-terpinene (85.1%, 22,318-2), 2.4 Antimicrobial testings
γ-terpinene (97.2%, 22,319-0), terpinolene (90.2%,
The essential oil of marjoram, the target compounds
W30,460-3-K), myrcene (90.0%, 276200), (S)-(-)-limo-
and the reference compound were prepared as 20%
nene (95.0%, W504505), and tetracycline hydrochlo-
solutions of ethanol and dissolved in a 0.9% NaCl so-
ride (achromycine hydrochloride – 25g, T3383-25G)
lution. As test microorganisms (colony-forming units
are products from Sigma-Aldrich Austria Co., Vienna.
per cm3 = cfu/cm3), Gram-positive bacteria Staphylo-
The essential marjoram oil sample from Albania
ATCC 6538P (1.5x1013) and Enterococ-
(800522) was obtained by hydrodistillation from the
(clinical isolated, 1x1013), Gram-negative
ﬂowering tops of Origanum majorana
L. with physi-
bacteria Escherichia coli
ATCC 8739 (1x1011), Pseudo-
co-chemical data: rotation +17.7°, density: 0.892,
G 28 (2x1013), Klebsiella pneumoni-
refraction: 1.4754; and the following reference com-
(clinical isolated, 2x1013), Proteus vulgaris
pounds as terpinen-4-ol (99.1%, 800760), α-terpineol isolated, 6x1013) and Salmonella enterica
(82%, 800761), linalool (96%, 800506), linalyl acetate
(clinical isolated, 3x1013) as well as the yeast
(96%, 800507), p
-cymene (97.0%, 800613), α-pinene Candida albicans
ATCC 10231 (1.9x1010) – all products
(97%, 800079), β-pinene (95%, 800140), β-caryophyl-
from the National Bank of Industrial Microorganisms
lene (97.5%, 800173), eugenol (98.3%, 800316) were
and Cell Cultures, Soﬁa, Bulgaria – were used.
purchased from Kurt Kitzing Co., Wallerstein. Ciprox-
The antimicrobial activity was studied by two well-
in® 500 mg tablets (1 tablet = 582 mg ciproﬂoxacine
established and optimised methods as follows: agar
hydrochloride/water) were bought from Bayer Austria
diffusion disc method using 6 mm paper discs and
Co., Vienna and Lidaprim® infusion bottle (250 mg
quantities of 6.0 μL of the sample. After cultivation of
containing 0.8 g sulfametrol and 0.16 g trimethoprim)
the bacteria and the yeast at 37 °C for 24 h the diam-
eter of the inhibition zone (IZ) was measured. Agar serial tube dilution method results in a minimum in-
2.2 GC analysis
hibitory concentration (MIC) as follows: The essential
GC/FID analyses were carried out using a GC-14A with
oil, pure and reference compounds were added to a
split/splitless injector, FID and C-R6A-Chromatopac in-
saline solution (0.5% NaCl in distilled water) contain-
tegrator (Shimadzu, Japan), a GC-3700 with FID (Var-
ing 1.0% (v/v) Tween 80 at the appropriate volumes
ian, Germany) and C-R1B-Chromatopac integrator
to produce ﬁnal concentrations of the samples in
(Shimadzu). The carrier gas was hydrogen (ﬂow rate:
the range of 1–1000 ppm. Petri dishes were inoculat-
1 mL/min); injector temperature 250 °C; detector tem-
ed by pipetting 6.0 μL of the serial diluted samples,
perature, 320 °C. The temperature programme was:
the reference compounds (the tablets of Ciproxin®
40 °C/5 min to 280 °C/5 min, with a heating rate of
were added as solution in saline at a quantity of
6 °C/min. The columns were 30 m x 0.25 mm bonded
300 μg) and 0.1 cm3 of the desired culture on paper
DB-5MS fused silica, with a ﬁlm thickness of 0.50 μm discs (6 mm) and then incubated at 37 °C for 24 h.
(J & W Scientiﬁc Corp., USA) and 30 m x 0.32 mm
bonded Stabilwax, with a ﬁlm thickness of 0.50 μm 2.5 Olfactoric evaluations
(Restek, USA). Quantiﬁcation was achieved using
The essential marjoram oil from Albania was olfac-
peak area calculations, and compound identiﬁcation
torily evaluated by a professional perfumer and two
was carried out partly using correlations between re-
aroma chemists; the aroma described in Tab. 1
lated with odour impressions published [1, 24–29].
2.3 GC/MS analysis
3 Results and discussions
For GC/MS measurements a GC-17A with QP5050 (Shi-madzu, Japan), split/splitless injector and HP-Com-
The essential oil of Origanum majorana
L. from Albania
paq data system (Hewlett-Packard, USA, Shimadzu
was olfactorily evaluated as follows: herbaceous, aromat-
GCMSsolution software) and a GC-17A with QP5000
ic-spicy, slightly terpenous, reminding of terpinen-4-ol.
(Shimadzu, Japan), split/splitless injector and Pen-
By means of gas chromatographic-spectroscopic
tium PC data system (HP, USA, Shimadzu class5k
analysis (GC/FID and GC/MS) of the essential marjo-
software) were used. The carrier gas was helium (flow
ram oil 28 constituents were identiﬁed. The chemical
rate: 1 mL/min); injector temperature 250 °C; interface
composition conﬁrms the authenticity of the sample
heating at 300 °C, ionsourceheating at 200 °C, EI-
by comparing retention indices with those of literature
mode was 70 eV, and the scan range was 41–450 amu.
data mentioned in Tab. 1
as well as scientiﬁc publica-
* non-polar/polar column# in relative % peak area using GC-FID with a non-polar column (mean value of 3 analyses)
§ in accordance to published data [1, 24–29]
of high antimicrobial active, phenolic compounds euge-
Tab. 1: Composition and olfactoric evaluation of the marjoram essential oil from
nol and the synthetic antibi-
Tab. 1: Zusammensetzung und olfaktorische Beurteilung des ätherischen Majo-
ride and Ciproxin® (these
ranöls aus Albanien.
products and Lidaprim® are known to have no effects
tions and own databases. The main compounds were
against the yeast C. albicans
) prove the effectivity of
as follows (concentration higher than 3.0%, calculated
the chosen testing methods, while Lidaprim® (no ef-
as relative % peak area using GC/FID with a non-polar
fects against P. aeruginosa
and K. pneumoniae
) did not
column, see Tab. 1
): terpinen-4-ol (21.33%), trans
show the antimicrobial activity as expected .
nene hydrate (15.53%), γ-terpinene (14.00%), α-terpi-
Antimicrobial tests with pure compounds, identiﬁed as
nene (8.88%), sabinene (8.27%), cis
main or minor constituents of the essential marjoram
(3.80%), α-terpineol (3.26%) and terpinolene (3.20%).
oil, such as α- and γ-terpinene, terpinolene, terpinen-
Using olfactoric data of previously published refer-
4-ol, α-terpineol, linalool, linalyl acetate, p
ences (see references cited in Tab. 1
) the character-
myrcene, limonene, α- and β-pinene, sabinene hy-
istic odour of the marjoram sample can be especially
drate (mixture of cis
- and trans
-sabinene hydrate) and
correlated to α-thujene, α-terpinene, γ-terpinene and caryophyllene showed the following antimicrobial ac-
bicyclogermacrene (herbaceous notes), sabinene,
tivity (see Tab. 2
): Only the monoterpene alcohols ter-
α-phellandrene, β-phellandrene, cis
- and trans
pinen-4-ol, α-terpineol and linalool were moderately
nene hydrate, cis
-sabinene hydrate acetate, β-caryo-
to highly effective against all strains of Gram-positive
phyllene, α-humulene, bicyclogermacrene and caryo-
and Gram-negative bacteria as well as the yeast, simi-
phyllene oxide (aromatic-spicy notes), α- and β-pi-
larly to the marjoram sample. Therefore, these three
nene, α- and γ-terpinene, terpinolene and terpinen-4-ol constituents are of basic importance for the excellent
1.5x1013 cfu/cm3 1x1013 cfu/cm3 1x1011 cfu/cm3 2x1013 cfu/cm3 6x1013 cfu/cm3 2x1013 cfu/cm3 3x1012 cfu/cm3 1.9x1010 cfu/cm3
Tab. 2: Antimicrobial activities of the essential marjoram oil from Albania, some main and minor compounds
as well as reference compounds.
Tab. 2: Antimikrobielle Aktivität des ätherischen Majoranöls, einiger Haupt- und Nebenkomponenten sowie der
antimicrobial properties of the essential O. majorana
oil from Albania. All other tested compounds (without
The authors acknowledge the valuable help of Prof.
γ-terpinene against S. aureus
and myrcene against
Dr. Heinz Schilcher
, Emeritus of the Freie Universität
both Gram-positive bacteria) were found to have a
Berlin, in the field of antimicrobial test methods.
medium to high effectivity against the Gram-positive bacteria and the yeast C. albicans
(only myrcene did
not show any activity) and therefore may act with the above-mentioned monoterpene alcohols in a syner-
 Jirovetz L.
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In conclusion, we can report that the essential Origa-
 United States Department of Agriculture, USDA
oil from Albania of our interest was es-
Agricultural Research Service. ARS, National Ge-
pecially rich in terpinen-4-ol, trans
netic Resources Program. Germplasm Resources
and γ-terpinene, and showed medium to high antimi-
Information Network (GRIN) [Online-Datenbank],
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National Germplasm Resources Laboratory,
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(29 January 2008).
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19.07.2008 / Menschen und Leben / Seite 17 Zwischen Kiez und Kino Kida Ramadan – ein Schauspieler, der aus dem Alltag kommt Er spricht deutsch, arabisch, türkisch und kurdisch und spielt in seiner Freizeit gerne Boules: Filmkünstler Kida Ramadan (31) am Paul-Lincke-Ufer in Berlin-Kreuzberg. Foto: ND/Ulli Winkler Knallhart« steht in arabischen Buchstaben auf dem T-Shirt, das Ki
UDC 658.155.001.18 Ishchenko Mykola Ivanovych, Ph.D. (Economics), Member of Executive Board, Director of Mining Division of METINVEST HOLDING LLC DETERMINATION OF ENTERPRISE EXCESSIVE LIABILITIES AT ITS FINANCIAL AND ECONOMIC PERFORMANCE EVALUATION Problem definition . Different features and ratios are used for defining financial and economic performance of the enterpr