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Chemical compounds and antimicrobial activity of petitgrain (Citrus aurantium L. var. amara) essential oil


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Introduction: Due to its low cost and easy availability on the market, the petitgrain oil is commonly used in food, cosmetics, and aromatherapy. Objective: The examination of chemical composition and antibacterial activity of commercial petitgrain oil. Methods: Identification of chemical components of the petitgrain oil was performed by gas chromatography (GC). The minimum inhibitory concentrations (MIC) and minimum bactericidal/fungicidal concentrations (MBC/MFC) were determined using macrodilution method for the reference strains of bacteria and fungi. Results: Twenty components were identified. The petitgrain oil contained mostly oxygenated monoterpene hydrocarbons (98.01%), and the main components included linalyl acetate (48.06%) and linalool (26.88%). The MIC/MBC of the petitgrain oil for bacteria was in the range of 0.63-5.0/1.25-5.0 mg/ml and for fungi in the range of 1.25-40/5.0-80 mg/ml. Conclusion: The petitgrain oil had higher antibacterial activity than antifungal activity. Bacillus subtilis among the tested bacteria and Aspergillus niger and Penicillium expansum among the fungi were found to be highly inhibited by the petitgrain oil.
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Vol. 63 No. 4 2017
From Botanical to Medical Research
DOI: 10.1515/hepo-2017-0021
Chemical compounds and antimicrobial activity
of petitgrain (Citrus aurantium L. var. amara)
essential oil
1Department of Biotechnology, Microbiology and Food Evaluation
Warsaw University of Life Sciences – SGGW
Nowoursynowska 159c
02-776 Warsaw, Poland
2Department of Vegetable and Medicinal Plants
Warsaw University of Life Sciences – SGGW
Nowoursynowska 159c
02-776 Warsaw, Poland
* corresponding author: e-mail:
Introduction: Due to its low cost and easy availability on the market, the petitgrain oil is commonly used
in food, cosmetics, and aromatherapy.
Objective: The examination of chemical composition and antibacterial activity of commercial petitgrain oil.
Methods: Identification of chemical components of the petitgrain oil was performed by gas chromatography
(GC). The minimum inhibitory concentrations (MIC) and minimum bactericidal/fungicidal concentrations
(MBC/MFC) were determined using macrodilution method for the reference strains of bacteria and fungi.
Results: Twenty components were identified. The petitgrain oil contained mostly oxygenated monoter-
pene hydrocarbons (98.01%), and the main components included linalyl acetate (48.06%) and linalool
(26.88%). The MIC/MBC of the petitgrain oil for bacteria was in the range of 0.63–5.0/1.25–5.0 mg/ml
and for fungi in the range of 1.25–40/5.0–80 mg/ml.
Conclusion: The petitgrain oil had higher antibacterial activity than antifungal activity. Bacillus subtilis
among the tested bacteria and Aspergillus niger and Penicillium expansum among the fungi were found to
be highly inhibited by the petitgrain oil.
Key words: petitgrain, essential oil, chemical composition, antimicrobial activity
Received: 2017-03-21
Accepted: 2017-08-30
Herba Pol 2017; 63(4): 18-25
Chemical compounds and antimicrobial activity of petitgrain (Citrus aurantium L. var. amara) essential oil
Vol. 63 No. 4 2017
Citrus aurantium L. var. amara of the Rutaceae
family, commonly known as bitter (“sour”) orange, is
regularly cultivated in the Mediterranean area and in
Central and South America [1]. Composition of es-
sential oils from bitter orange is not the same and it
depends, to a large extent, on the geographic origin
and parts of the plant (zest, fruit, and owers) [2].
Essential oil from fruit zest is obtained through the
cold-pressed extraction method and it is referred to
as “cold-pressed essential oil”. e most expensive
in production is “Neroli oil”, which is obtained from
petals using steam distillation or hydrodistillation
method [1]. Petitgrain bigarade oil is obtained by
steam distillation [3] or hydrodistillation of the leaves
and twigs from pruning of the trees at dierent times
in the year [4]. Among essential oils obtained from
bitter orange, petitgrain is the lowest-priced oil [5].
e largest producer of the petitgrain oil is Paraguay,
followed by Egypt, Spain, France, and Italy [6]. e
total production of this essential oil is estimated at
260 tons per annum [7]. Petitgrain oil is also recom-
mended for aromatherapy, particularly for the antide-
pressant treatment, since it relieves anxiety, agitation,
stress, and challenging behaviors [8]. Due to its low
cost, and easy availability in the market, this essential
oil is commonly used in the production of marma-
lades and in avoring of some types of beers [9].
e study was aimed at the examination of the
chemical composition and antimicrobial activity
of commercial petitgrain oil against four bacterial
strains, including the pathogens transferred via food
and four fungi strains found in food.
Essential oil
In this research, petitgrain essential oil (from Cit-
rus aurantium L. var. amara) was used. e oil was
purchased from the Pollena Aroma Company, Po-
land (commercial producer of plant essential oils and
aromatic substances), from three dierent batches.
Quality of the essential oil was ascertained to be
higher than 98% pure. e oils were stored in tightly
closed dark vials at 4°C until further tests and analysis.
Identication of chemical components
Gas chromatography analysis was performed us-
ing a Hewlett Packard 6890 gas chromatograph
equipped with aame ionization detector (FID) and
capillary, polar column HP 20M (25 m × 032 mm,
0.3 µm lm thickness). e analysis was performed
following the temperature program: oven tempera-
ture isotherm at 60°C for 2 min, then from 60°C to
220°C at a rate of 4°C/min and held at 220°C for
5 min. Injector and detector temperatures were set
at 220°C and 260°C, respectively. e carrier gas
(He) ow was 1.1 ml/min. e split ratio was 1:70.
Atotal of 0.1 µl of pure essential oil was manually
injected. Component identication was performed
by comparison of their retention times with those of
pure authentic samples and by their linear retention
indices (RI) relative to the series of n-hydrocarbons
(C7–C30), under the same conditions. Retention in-
dices of compounds were also compared with those
reported in the literature. e percentage composi-
tion of the oils was computed by the normalization
method from the GC peak areas, without the use of
correction factors. All the analyzes were performed
in triplicate.
Microorganisms and inoculum preparation
e reference strains Bacillus subtilis ATCC 6633,
Staphylococcus aureus ATCC 25923, Escherichia
coli ATCC 25922, Salmonella ser. Enteritidis ATCC
13076, Saccharomyces cerevisiae ATCC 9763, Can-
dida krusei ATCC 14243, Aspergillus niger ATCC
9142, and Penicillium expansum ATCC 7861 were
obtained from the culture collection (Division of
Food Biotechnology and Microbiology, Warsaw
University of Life Sciences, Warsaw, Poland). e
strains were stored in 20% glycerol at -80°C in
e frozen subcultures of bacteria were trans-
ferred to tubes containing 5 ml of sterile nutrient
broth (BTL). Aer incubation the test cultures of the
bacteria strains were separately inoculated on slants
of nutrient agar (BTL) and incubated at 37°C for
24 hours. Bacterial inocula were prepared in asterile
saline (0.85% NaCl) (w/v) solution with the quantity
corresponding to 0.5 McFarland (~1×108 cfu/ml)
and diluted to ~1×107 cfu/ml.
e yeasts were separately inoculated on Sab-
ouraud Agar slopes (SA, Merck) and were incubated
at 28°C for 48 hours. Yeast inocula were prepared
in asterile saline solution. e density of the yeast
suspension were measured with a hemocytometer
and diluted as necessary with sterile saline to obtain
the inoculum density of ~1×106 cfu/ml. e molds
were separately inoculated on Potato Dextrose Agar
Gniewosz, K. Kraśniewska, O. Kosakowska, K. Pobiega, I. Wolska
slopes (PDA, BTL) and incubated at 28°C for 7 days.
Slopes were ooded with 1 ml of phosphate-buered
saline (PBS) containing 0.05% Tween 80 and gently
probed to dislodge the conidia [10]. Aer the set-
tling of larger particles, suspensions were adjusted
by nephelometry and diluted as necessary to obtain
the inoculum density of ~1×106 cfu/ml.
Determination of MIC and MBC/MFC
e antimicrobial activity of the petitgrain oil was
assayed with dilution broth method according to
the EUCAST guidelines [11]. Double series of petit-
grain oil dilutions ranging from 0.078 to 80 mg/ml
(v/v) were prepared for the respective bacteria in
Mueller-Hinton Broth (MHB, Merck) and for fungi
in Sabouraud Broth (SB, Merck) [12] supplemented
with 0.002% (v/v) of Tween 80 (Sigma-Aldrich).
Anegative control (medium and inoculum) was ad-
ditionally prepared for each experimental series. Fi-
nally, each test tube contained 2 ml of medium with
the appropriate concentration of petitgrain oil and
0.1 ml of inoculum.
e 100 µl of inoculum containing approxi-
mately 1 × 107 cfu/ml of bacteria were transferred
in MBH so that the nal concentration in each
tube was ~ 5.0 × 105 cfu/ml and 100 µl of the in-
oculum containing approximately 1×106 cfu/ml of
fungi were added in SB, so that the nal inoculum
concentrations were in the range of 0.4 × 104
5.0 × 104 cfu/ml [10]. Apositive control (contain-
ing inoculum without petitgrain oil) was includ-
ed in each series. Aer incubation at appropriate
temperature and time (bacteria at 37°C for 24 h,
yeast at 28°C for 48 hours, and molds at 28°C for
72 hours), the MIC was checked. e MIC is the
lowest concentration of petitgrain oil at which the
bacteria and fungi failed to grow, no visible chang-
es were detected in the broth medium.
In order to evaluate the minimum bacteri-
cidal/fungicidal concentration (MBC/MFC) of
petitgrain oil, 100 µl of each culture from tubes
in which microbial growth was not observed was
spread on Mueller-Hinton Agar (MHA, Merck)
for bacteria and on Sabouraud Agar (SA, Merck)
plates for fungi. Plates were incubated at appropri-
ate temperature and time (bacteria at 37°C for 24 h,
yeast at 28°C for 48 hours, and molds at 28°C for
72 hours).e complete absence of growth of bac-
terial or fungal colonies on the agar surface at the
lowest concentration of petitgrain oil was dened
as the minimum bactericidal concentration (MBC)
or minimum fungicidal concentration (MFC). e
evaluation of MIC and MBC/MFC was carried out
in triplicate.
Ethical approval: e conducted research is not re-
lated to either human or animal use.
Characterization of essential oil
e chemical composition of petitgrain oil analyzed
using the GC/FID is presented in table 1. Twenty
chemical compounds were determined in the ana-
lyzed essential oil. e percentage content of its
components was 98.90%. e highest percentage of
content characterized was of monoterpenoid com-
pounds and oxygenated monoterpene hydrocar-
bons, which formed 98.01% of the oil. e majority
of the identied compounds belong to the oxygen-
ated monoterpene hydrocarbons: linalyl acetate
(48.06%), linalool (26.88%), α-terpineol (5.74%),
geranyl acetate (3.92%), geraniol (3.05%), and gera-
nial (2.44%).
Based on chemical analysis of petitgrain oil pre-
sented in this study and several other previous ex-
aminations, it can be concluded that oxygenated
monoterpenes are present at higher concentrations
than monoterpene hydrocarbons in the oil. eir
content constitutes over 90% of total composition of
petitgrain oil. e major components of petitgrain oil
are linalyl acetate, linalool, α-terpineol, and geranyl
acetate [7, 13]. Petitgrain oils originating from vari-
ous areas of the world dier in the content of main
components which may be due to dierent plant gen-
otype, climate, and soil conditions [13, 14]. Linalool
was the predominant among the main components
of the Tunisian oil (62.57−22.35%), with lower con-
tent of linalyl acetate (25.38–5.64%) and α-terpineol
(3.0–15.69%) [15]. e petitgrain oil originating from
Greece contained 88.09% oxygenated monoterpenes,
mostly linalool (58.21%). Apart from these, the Greek
petitgrain oil contained neryl acetate and trans-β-
ocimene, which were not determined in the com-
position of commercial petitgrain oil. On the other
hand, Sicilian petitgrain oil was characterized with
higher values of linalyl acetate (0.3–73.1%) and lower
linalool content (8.7–16.7%) [16]. Petitgrain oil from
Turkey has ahigh content of oxygenated compounds
(89.6%) with linalyl acetate (50.1%) and linalool
(24.8%) being the main components [3]. e chemi-
cal composition of petitgrain oil is further inuenced
Chemical compounds and antimicrobial activity of petitgrain (Citrus aurantium L. var. amara) essential oil
Vol. 63 No. 4 2017
by the procedure and the time of extraction from the
plant raw material [15]. Ellouze and Abderrabba [15]
determined that during hydrodistillation of essential
oil from C. aurantium leaves for 180 min, adecline in
monoterpene content was observed (89.93–47.32%
in 15 min and 165 min, respectively), while the ses-
quiterpene content increased. Ellouze et al. [4] have
found the eect of seasonal variations of leaves col-
lection on chemical composition of essential oils.
e essential oils obtained from the fresh leaves of
C. autrantium L. ssp. aurantium, which were gathered
during January noted 14 components only, while the
July one presented 35 among the 46 identied com-
Antimicrobial activity of petitgrain oil
e evaluation of antimicrobial activity of petitgrain
oil was conducted in compliance with the standard-
ized techniques, determining the minimum inhibi-
tory concentrations (MIC) that inhibits the growth
of microbes and minimum bactericidal and fungi-
cidal concentrations (MBC/MFC). e assessment
of the eciency of petitgrain oil was conducted for
the reference test strains of bacteria and fungi.
e MIC and MBC/MFC of petitgrain oil are
presented in table 2. e highest inhibitory activ-
ity of petitgrain oil was determined for bacteria,
including Gram-positive bacteria. e MIC values
Table 1.
Gas chromatographic composition (% peak area) of petitgrain (Citrus aurantium L. var. amara) essential oil
Compound RIaRIbRI b range Mean ±SDc
α-Pinene 1028 1025 1008–1039 0.16±0.02
β-Pinene 1113 1110 1085–1130 1.14±0.12
Sabinene 1124 1120 1098–1140 0.27±0.37
β-Myrcene 1166 1161 1140–1175 1.62±0.15
Limonene 1203 1198 1178–1219 1.40±0.12
1,8 Cineole 1209 1211 1186–1231 0.04±0.03
γ-Terpinene 1248 1245 1222–1266 1.29±0.10
p-Cymene 1273 1270 1246–1291 0.06±0.01
α-Terpinolene 1278 1282 1261–1300 0.23±0.02
Linalool 1540 1543 1507–1564 26.88±0.88
Linalyl acetate 1557 1554 1532–1570 48.06±1.05
β-Caryophyllene 1593 1588 1570–1685 0.96±0.04
Isoborneol 1657 1659 1635–1675 0.09±0.01
α-Terpineol 1681 1694 1659–1724 5.74±0.11
Borneol 1687 1699 1653–1728 0.35±0.01
Geranial 1722 1725 1680–1750 2.44±0.12
Geranyl acetate 1731 1751 1728–1772 3.92±0.23
Nerol 1795 1794 1752–1832 1.36±0.31
Geraniol 1826 1839 1795–1865 3.05±0.04
Caryophyllene oxide 1955 1986 1936–2023 0.08±0.02
Identied components [%] 99.14
Grouped components
Monoterpene hydrocarbons 6.17
Oxygenated monoterpene
Sesquiterpene hydrocarbons 0.96
Oxygenated sesquiterpene
Notes: RIa – etention index relative on HP-20M capillary column; RIb – average retention indices on polar column reported by Babushok et al. [13],
RIb range – range of retention indices on polar column reported by Babushok et al. [13]; SDc – standard deviation, n=3
Gniewosz, K. Kraśniewska, O. Kosakowska, K. Pobiega, I. Wolska
of petitgrain oil for B. subtilis and S. aureus bacte-
ria remained in the range of 0.63–1.25 mg/ml, while
the MBC values were 1.25–5.0 mg/ml. e Gram-
negative bacteria strains of E. coli and S. ser. Enter-
itidis were less susceptible to the eect of petitgrain
oil. e MIC and MBC values remained in the range
between 2.5 and 5.0 mg/ml.
e weakest antimicrobial activity of petitgrain
oil was observed against fungi. e MIC of petit-
grain oil for the inhibition of P. expansum and A. ni-
ger mold was 1.25 mg/ml. In the case of S. cerevisiae
yeast, the MIC and MFC values were 2.5 mg/ml and
5.0 mg/ml, respectively. e weakest fungicidal ef-
fect of petitgrain oil was observed for C. krusei.
e antimicrobial activity of essential oils is typi-
cally linked to their main components [17]. Petitgrain
oil is characterized with high content of oxygenated
monoterpenes, mostly linalyl acetate (48.06%) and
linalool (26.88%) (tab. 1). e mechanism of anti-
microbial activity of these compounds stems from
their lipophilic character, creating strong anity to
plasma membranes of microorganisms and strong
toxicity [18]. Linalyl acetate and limonene preferen-
tially divide from an aqueous phase and enter into
the plasma membrane structures. Accumulation of
linalyl acetate and limonene in the membrane dam-
ages the composition and structure of plasma mem-
brane, causing the increase of plasma membrane
uidity and leakage of intracellular molecules from
the cell. Blaskó et al. [18] conrmed that these dis-
turbances in plasma membrane determine the cell
death of Candida albicans. Similar eect is attributed
to 1,8-cyneol, which modies the characters of cel-
lular membranes by rupturing the hydrogen bonded
network at the membrane-water interface. Mono-
terpene alcohols (α-terpineol and 1,8-cineole) have
asimilar eect on microbial cells, i.e., they interact
with plasma membranes [12]. At relatively low con-
centrations, such interactions may lead to changes
like respiratory inhibition and altered permeability
[19], and at higher concentrations they may lead to
acomplete loss of homeostasis, plasma membrane
damage, and death. Monoterpene alcohols are con-
sidered to be ecient against microbes due to their
relatively high water solubility and the presence of
the alcohol moiety [19].
ere are published data from previous studies
regarding the antimicrobial activity of essential oils
produced from owers and fruits of Citrus auran-
tium L. var. amara, but no data is available on the
antimicrobial activity of petitgrain oil. Ellouze et
al. [4] registered that petitgrain oils were most ef-
fective against Gram-positive bacteria (S. aureus
and L. monocytogenes were more sensitive to the
essential oils), and weak against Gram-negative
strains. Essential oil from bitter orange owers ex-
amined by Hsouna et al. [17] inhibited the growth
of both Gram-positive and Gram-negative bacteria
in the concentration range from 0.312–2.5 mg/ml
and 0.625–2.5 mg/ml, respectively. e fungistatic
activity of this essential oil was even stronger. e
MIC for molds of the genus Aspergillus was in the
range of 0.078–1.25 mg/ml and for Fusarium, it
was 0.156–1.25 mg/ml. A comparison of MIC val-
ues of both the essential oils from Citrus aurantium
L. var. amara demonstrated that antimicrobial activ-
ity of the oil from owers was stronger than the oil
from leaves and twigs (petitgrain) of the plant. e
ower oil contained considerably higher amounts of
monoterpene hydrocarbons (36.2%), sesquiterpene
hydrocarbons (4.1%), and oxygenated sesquiterpene
hydrocarbons (26.2%), which were present in the
examined petitgrain oil at 6.17, 0.96, and 0.08%, re-
spectively. Strong fungicidal activity is also exhibited
Table 2.
e antimicrobial activity of petitgrain essential oil
Microorganism MIC MBC
B. subtilis ATCC 6633 0.63 1.25
S. aureus ATCC25923 1.25 5
Bacteria E. coli ATCC 25922 2.5 5
S. ser. Enteritidis ATCC 13076 5 5
S. cerevisiae ATCC 9763 2.5 5
A. niger ATCC 9142 1.25 40
Fungi P. expansum ATCC 7861 1.25 80
C. krusei ATCC 14243 40 >80
Chemical compounds and antimicrobial activity of petitgrain (Citrus aurantium L. var. amara) essential oil
Vol. 63 No. 4 2017
by limonene and (E)-nerolidol, which were present
in the ower essential oil at concentrations 27 and
17.5%, respectively. In the petitgrain oil, limonene
was present only at aconcentration of 1.4%.
e unequal eect of the oil is linked to the dif-
ference in its composition of active compounds.
Monoterpenes might be responsible for the anti-
fungal eect of the petitgrain oil. Hammer et al.
[19] demonstrated monoterpene activity toward
yeasts and lamentous fungi. In the study, it was
determined that Candida albicans and C. tropi-
calis are characterized by high susceptibility to
monoterpene mixtures. High antifungal activity is
also exhibited by terpinen-4-ol, α-pinen, β-pinen,
1,8-cyneol, linalool, and 4-terpineol. is mixture
of terpenoid metabolites inhibits the growth of
dermatophytes, such as Trichophyton mentagro-
phytes, T. rubrum, and Microsporum gypseum, and
also the lamentous fungi Aspergillus niger and
A. avus [20].
Essential oils are natural plant products containing
mixture of components and thus having multiple an-
timicrobial properties. In the petitgrain essential oil,
the GC/FID analysis allowed to identied 20 com-
pounds. e major components of essential oil was
linalyl acetate (48.06%) and linalool (26.88%).
e results of the study may suggest eective an-
timicrobial activity of petitgrain essential oil. It was
found that tested oil eectively inhibits the growth
of Gram-positive bacteria. Less sensitive to the in-
hibitory activity were Gram-negative bacteria and
fungi. is nding also highlights the potential use
of the petitgrain essential oil as inhibitors of food
spoilage and pathogenic microorganisms.
e work was funded as statutory research by the
Department of Biotechnology, Microbiology and
Food Evaluation, Faculty of Food Science, Warsaw
University of Life Sciences-SGGW.
Conict of interest: Authors declare no conict of interest.
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Skład chemiczny iaktywność przeciwdrobnoustrojowa olejku eterycznego
petitgrain (Citrus aurantium L. var. amara)
1Katedra Biotechnologii, Mikrobiologii iOceny Żywności
Szkoła Główna Gospodarstwa Wiejskiego wWarszawie
ul. Nowoursynowska 159C
02-776 Warszawa
2Katedra Roślin Warzywnych iLeczniczych
Szkoła Główna Gospodarstwa Wiejskiego wWarszawie
ul. Nowoursynowska 159C
02-776 Warszawa
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any wprodukcji żywności, kosmetyków iaromaterapii.
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16. De Pasquale F, Siragusa M, Abbate L, Tusa N, De
Pasquale C, Alonzo G. Characterization of ve
sour orange clones through molecular markers
and leaf essential oils analysis. Sci Hort 2006;
17. Hsouna AB, Hamdi N, Halima NB, Abdelka S.
Characterization of essential oil from Citrus au-
rantium L. owers: antimicrobial and antioxidant
activities. J Oleo Sci 2013; 62:763-772.
18. Blaskó A, Gazdag Z, Gróf P, Máté G, Sárosi S, Kr-
isch J et al. Eects of clary sage oil and its main
components, linalool and linalyl acetate, on the
plasma membrane of Candida albicans: an in vivo
EPR study. Apoptosis 2016; 22(2):175-187.
19. Hammer KA, Carson CF, Riley TV. Antifungal
activity of the components of Melaleuca alterni-
folia (tea tree) oil. J Appl Microbiol 2003; 95:853-
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e role of structure and molecular properties of
terpenoids in determining their antimicrobial ac-
tivity. Flavour Frag J 1999; 14: 322-332.
Chemical compounds and antimicrobial activity of petitgrain (Citrus aurantium L. var. amara) essential oil
Vol. 63 No. 4 2017
Cel: Zbadano skład chemiczny iaktywność przeciwdrobnoustrojową handlowego olejku petitgrain.
Metodyka: Identyfikację chemicznych składników olejku petitgrain wykonano przy użyciu chromatografii
gazowej (GC). Minimalne stężenia hamujące (mic) oraz minimalne stężenia bakteriobójcze/grzybobójcze
(MBC/MFC) zostały oznaczone metodą makrorozcieńczeń wobec referencyjnych szczepów bakterii igrzy-
Wyniki: Zidentyfikowano dwadzieścia kompotentów. Olejek petitgrain zawierał najwięcej utlenionych wę-
glowodorów monoterpenowych (98.01%), agłównymi składnikami były: octan linalilu (48,06%) ilinalool
(26,88%). MIC/MBC olejku petitgrain wobec bakterii były wgranicach 0,63–5,0/1,25−5,0 mg/ml, awobec
grzybów były wzakresie 1,25–40/5,0–80 mg/ml.
Wnioski: Olejek petitgrain miał większą aktywność przeciwbakteryjną niż przeciwgrzybiczną. Spośród ba-
danych bakterii Bacillus subtilis, aspośród grzybów Aspergillus niger iPenicillium expansum były najsilniej
hamowane przez olejek petitgrain.
Słowa kluczowe: petitgrain, olejek eteryczny, skład olejku eterycznego, aktywność przeciwdrobno-
... Linalyl acetate, linalool, α-terpineol, geranyl acetate, geraniol, and geranial as oxygenated monoterpene hydrocarbons were primarily identified in petitgrain oil of C. aurantium var. amara [12], whereas limonene was present only at a concentration of 1.4%. ...
... Terpinen-4-ol, α-pinene, β-pinene, 1,8-cyneol, linalool, and 4-terpineol and their mixture have been shown to have potent antifungal activity [12,43,44]. The most abundant compounds in Tunisian oil were linalool with lower amounts of linalyl acetate and α-terpineol [45]. ...
... Although cis-β-terpineol, D-limonene, 4-carvomenthenol, and linalool were the main compounds in petitgrain EO in the present study, the compounds of linalyl acetate, linalool, α-terpineol, and geranyl acetate [12,18,63] were the main compounds in petitgrain EO, which exhibited good antibacterial and antifungal activity, especially against Bacillus subtilis, Aspergillus niger, and Penicillium expansum, whereas the weakest fungicidal effects were observed for Candida krusei [12]. A mixture of terpenoid containing terpinen-4-ol and linalool exhibited high antifungal activity against Trichophyton mentagrophytes, T. rubrum, Microsporum gypseum, A. niger, and A. flavus [43]. ...
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In the present work, essential oils (EOs) extracted from different parts of sour orange Citrus aurantium (green leaves/twigs, small branches, wooden branches, and branch bark) were studied through gas chromatography coupled with mass spectrometry (GC/MS). Furthermore, the EOs in the amounts of 5, 10, 15, 20, and 25 µL were studied for their antibacterial activity against three pathogenic bacteria, Agrobacterium tumefaciens, Dickeya solani, and Erwinia amylovora. The main EO compounds in the leaves/twigs were 4-terpineol (22.59%), D-limonene (16.67%), 4-carvomenthenol (12.84%), and linalool (7.82%). In small green branches, they were D-limonene (71.57%), dodecane (4.80%), oleic acid (2.72%), and trans-palmitoleic acid (2.62%), while in branch bark were D-limonene (54.61%), γ-terpinene (6.68%), dodecane (5.73%), and dimethyl anthranilate (3.13%), and in branch wood were D-limonene (38.13%), dimethyl anthranilate (8.13%), (-)-β-fenchol (6.83%), and dodecane (5.31%). At 25 µL, the EO from branches showed the highest activity against A. tumefaciens (IZ value of 17.66 mm), and leaves/twigs EO against D. solani and E. amylovora had an IZ value of 17.33 mm. It could be concluded for the first time that the wood and branch bark of C. aurantium are a source of phytochemicals, with D-limonene being the predominant compound in the EO, with potential antibacterial activities. The compounds identified in all the studied parts might be appropriate for many applications, such as antimicrobial agents, cosmetics, and pharmaceuticals.
... Similar to the Poland petitgrain EO, 20 compounds were identified. The major components of petitgrain EO were linalyl acetate (48.06%) and linalool (26.88%) (Gniewosz et al., 2017). Linalool was the most abundant of the primary components of Tunisian oil (22.35-62.57%), ...
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The purpose of this study was to determine the chemical composition and biological properties of the citrus essential oils (EOs) derived from orange rinds (peels) of lemon (Citrus limon), wild orange (Citrus sinensis) from Brazil extracted by the cold pressed/expressed method, and leaves and twigs of petitgrain (Citrus aurantium) from Paraguay extracted by steam distillation. These food grade EOs were evaluated for their cytotoxic activity in breast, liver, and cervical cancer cells (MCF-7, HepG2 and HeLa) via MTT assay, antiproliferative activity via colony formation assay, and antimigratory activity via wound healing assay, and apoptosis via DNA fragmentation and morphology assessment. The major compounds found in lemon EO were D-limonene (66.75%), beta-pinene (12.82%), and gamma-terpinene (11.57%), totaling over 90% of the identified compounds. For wild orange, the only predominant compound was limonene (96.60%), and the rest, found in minor amounts, included alpha-pinene, bicyclohexane, beta-pinene, beta-myrcene, 3-carene, and o-cymene. For petitgrain EO, linalyl isobutyrate (51.76%) and linalool (26.86%) were mainly detected. Based on the MTT assay, petitgrain EO was the most effective against MCF-7, HepG2 and HeLa. However, wild orange EO was the most antiproliferative and antimigratory against all three cells using the anticolony formation assay and wound healing assay, respectively. The results showed that cell death is associated with the apoptotic process, with morphological hallmarks of apoptosis including membrane blebbing and DNA fragmentation. These findings imply that the three citrus EOs might be used as active components in functional food products for chemopreventive benefits.
... Olejek eteryczny z Citrus aurantium wykazuje aktywność względem Aspergillus fumigatus, hamując jego wzrost całkowicie przy 10-procentowym dodatku olejku do podłoża [16], co odpowiada wynikom otrzymanym w prezentowanej pracy w odniesieniu do Aspergillus niger. Dawka powodująca działanie hamujące wzrost A. niger, jakie wykazał olejek eteryczny z gorzkiej pomarańczy C. aurantium, została określona na 1,25 mg/ml [48], co potwierdziły również wyniki badań innych autorów [17]. Różnice mogą wynikać z różnic gatunkowych pomiędzy grzybami użytymi w badaniach a grzybami wskazanymi przez innych autorów oraz z różnic w składzie badanych olejków eterycznych wynikających z różnych metod ekstrakcji czy sezonu uprawy cytrusów. ...
... The main ingredients of sour orange (petitgrain) oil were linalool, (was the main alcohol components), linalyl acetate, (which considered the major component of ester fraction), (limonene, βocimene, myrcene and β-pinene were the highest monoterpene hydrocarbons), α-terpineol, geranyl acetate, geraniol and geranial (Abd El-Rashid, 2005& Gniewosz et al., 2017. ...
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Two field experiments were conducted during two successive seasons of 2017/2018 and 2018/2019 at the Horticulture Research Station Farm in El-Quassassin, Ismailia Governorate, Egypt, to assay the effect of various sources of aromatic waters of peppermint, geranium, spearmint, rosemary and sour orange (petitgrain) on growth parameters, yield, chemical constituents and volatile oil production of garlic plants (Allium sativum L.). Results showed that the foliar application with aromatic waters gave significant response in various aspects compared with plants treated with distilled water (control) during two seasons. Garlic plants that sprayed with petitgrain aromatic water significantly enhanced plant growth and yield. Moreover, the highest values of chemical contents and antioxidant activity were recorded for the same treatment. On the contrary, this treatment recorded the lowest nitrate content for both seasons. Results showed non-significantly affect among sulphur content values. It is remarkable that spraying with peppermint recorded significant volatile oil production compared with untreated plants during two seasons.
... This study used the following ten commercially available essential oils (dr Beta, Pollena Aroma, Nowy Dwór Mazowiecki, Poland): anise, cinnamon, citronella, clove, geranium, lavender, limette, mint, rosemary and thyme. The composition of tested EOs was presented in Table 1, based on data obtained from previous studies analyzing the EOs' composition by gas chromatography with flame-ionization and mass spectroscopic detection (GC-FID-MS) [46][47][48][49][50][51][52] or data from European Pharmacopoeia [53]. Table 1. ...
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Modifying the composition of dental restorative materials with antimicrobial agents might induce their antibacterial potential against cariogenic bacteria, e.g., S. mutans and L. acidophilus, as well as antifungal effect on C. albicans that are major oral pathogens. Essential oils (EOs) are widely known for antimicrobial activity and are successfully used in dental industry. The study aimed at evaluating antibacterial and antifungal activity of EOs and composite resin material (CR) modified with EO against oral pathogens. Ten EOs (i.e., anise, cinnamon, citronella, clove, geranium, lavender, limette, mint, rosemary thyme) were tested using agar diffusion method. Cinnamon and thyme EOs showed significantly highest antibacterial activity against S. mutans and L. acidophilus among all tested EOs. Anise and limette EOs showed no antibacterial activity against S. mutans. All tested EOs exhibited antifungal activity against C. albicans, whereas cinnamon EO showed significantly highest and limette EO significantly lowest activity. Next, 1, 2 or 5 μL of cinnamon EO was introduced into 2 g of CR and microbiologically tested. The modified CR showed higher antimicrobial activity in comparison to unmodified one. CR containing 2 μL of EO showed the best antimicrobial properties against S. mutans and C. albicans, while CR modified with 1 μL of EO showed the best antimicrobial properties against L. acidophilus.
... According to Fathi et al. [61], MBICs for E. coli and K. pneumoniae to C. aurantium essential oil in the study were 100 mg/mL and 150 mg/mL, respectively. The antimicrobial activity of CAEO reported by Gniewosz et al. [62] was 1.25 mg/mL MIC and 5 mg/mL MBC for S. aureus, 2.5 mg/mL and 5 mg/mL in case of Saccharomyces cerevisiae, and 2.5 mg/mL and 5 mg/mL MBC with respect to S. cerevisiae. ...
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The main aim of the study was to investigate the chemical composition, antioxidant, antimicrobial, and antibiofilm activity of Citrus aurantium essential oil (CAEO). The biofilm profile of Stenotrophonomonas maltophilia and Bacillus subtilis were assessed using the mass spectrometry MALDI-TOF MS Biotyper and the antibiofilm activity of Citrus aurantium (CAEO) was studied on wood and glass surfaces. A semi-quantitative composition using a modified version was applied for the CAEO characterization. The antioxidant activity of CAEO was determined using the DPPH method. The antimicrobial activity was analyzed by disc diffusion for two biofilm producing bacteria, while the vapor phase was used for three penicillia. The antibiofilm activity was observed with the agar microdilution method. The molecular differences of biofilm formation on different days were analyzed, and the genetic similarity was studied with dendrograms constructed from MSP spectra to illustrate the grouping profiles of S. maltophilia and B. subtilis. A differentiated branch was obtained for early growth variants of S. maltophilia for planktonic cells and all experimental groups. The time span can be reported for the grouping pattern of B. subtilis preferentially when comparing to the Molecules 2020, 25, 3956 2 of 21 media matrix, but without clear differences among variants. Furthermore, the minimum inhibitory doses of the CAEO were investigated against microscopic fungi. The results showed that CAEO was most active against Penicillium crustosum, in the vapor phase, on bread and carrot in situ.
... The phytocompounds of C. aurantium essential oil of fruit peels have been reported to possess antioxidant, antimicrobial, antifungal, antiparasitic, and anti-inflammatory activities (Sonbol et al., 1995;Ramadan et al., 1996;Wei and Shibamoto, 2010). The antifungal activity of C. aurantium essential oil (CAEO) extracted from leaves, fruits and flowers have been reported against fungal pathogens like Penicillium expansum, Aspergillus niger, C. albicans, and C. krusei (Chintaluri et al., 2015;Gniewosz et al., 2017). However, there are no reports on the synergistic potential of C. aurantium essential oil (CAEO) against fungal pathogens. ...
In research the antioxidant and antimicrobial activity of water and ethanolic extracts of Scutellaria baicalensis root on quality of chicken meatballs during refrigerated storage were determined. Extracts were analyzed for identification and quantification of polyphenols, DPPH and antimicrobial activity. Three different treatments of meatballs were prepared: control (MB), with addition of water extract (MBWE) and with addition of ethanolic extract (MBEE). The water extract was characterized by lower phenolic content (235.86 mg/gdm) and lower DPPH (257.56 μmol Trolox/gdm) than ethanolic extract. The ethanolic extract was also characterized by higher antimicrobial activity in relation to selected strains of microorganisms. Oxidative changes in MBEE were significantly (p < 0.05) lower in comparison to MB. This was indicated by lower (by 50%) TBARS values and almost twice as long fat induction time. A higher content of polyphenol compounds in the ethanolic extract (especially baicalein) resulted in a more effective reduction of lipid oxidation.
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The effects of clary sage (Salvia sclarea L.) oil (CS-oil), and its two main components, linalool (Lol) and linalyl acetate (LA), on cells of the eukaryotic human pathogen yeast Candida albicans were studied. Dynamic and thermodynamic properties of the plasma membrane were investigated by electron paramagnetic resonance (EPR) spectroscopy, with 5-doxylstearic acid (5-SASL) and 16-SASL as spin labels. The monitoring of the head group regions with 5-SASL revealed break-point frequency decrease in a temperature dependent manner of the plasma membrane between 9.55 and 13.15 °C in untreated, in CS-oil-, Lol- and LA-treated membranes. The results suggest a significant increase in fluidity of the treated plasma membranes close to the head groups. Comparison of the results observed with the two spin labels demonstrated that CS-oil and LA induced an increased level of fluidization at both depths of the plasma membrane. Whereas Lol treatment induced a less (1 %) ordered bilayer organization in the superficial regions and an increased (10 %) order of the membrane leaflet in deeper layers. Acute toxicity tests and EPR results indicated that both the apoptotic and the effects exerted on the plasma membrane fluidity depended on the composition and chemical structure of the examined materials. In comparison with the control, treatment with CS-oil, Lol or LA induced 13.0, 12.3 and 26.4 % loss respectively, of the metabolites absorbing at 260 nm, as a biological consequence of the plasma membrane fluidizing effects. Our results confirmed that clary sage oil causes plasma membrane perturbations which leads to cell apoptosis process.
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Nowadays, use of alternative and complementary therapies with mainstream medicine has gained the momentum. Aromatherapy is one of the complementary therapies which use essential oils as the major therapeutic agents to treat several diseases. The essential or volatile oils are extracted from the flowers, barks, stem, leaves, roots, fruits and other parts of the plant by various methods. It came into existence after the scientists deciphered the antiseptic and skin permeability properties of essential oils. Inhalation, local application and baths are the major methods used in aromatherapy that utilize these oils to penetrate the human skin surface with marked aura. Once the oils are in the system, they remodulate themselves and work in a friendly manner at the site of malfunction or at the affected area. This type of therapy utilizes various permutation and combinations to get relief from numerous ailments like depression, indigestion, headache, insomnia, muscular pain, respiratory problems, skin ailments, swollen joints, urine associated complications etc. The essential oils are found to be more beneficial when other aspects of life and diet are given due consideration. This review explores the information available in the literature regarding therapeutic, medical, cosmetic, psychological, olfactory, massage aromatherapy, safety issues and different plants used in aromatherapy. All the available information was compiled from electronic databases such as Academic Journals, Ethnobotany, Google Scholar, PubMed, Science Direct, Web of Science, and library search.
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The essential oil constituents of the peel and leaf of Citrus aurantium L. (Rutaceae) grown in the north of Iran, were analyzed by GC/MS. Fourteen components representing 99.6% of the leaf oil were identified. The major compounds were linalool (39.4%), linalyl acetate (38.8%) and α-terpineol (7.2%). Twenty constituents consisting 99.4% of the peel oil were identified. The main ingredients were limonene (91.3%), β-myrcene (3.0%) and linalool (1.1%).
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Essential oils are natural materials widely used in many fields all over the world and have become an integral part of everyday life. There is increasing demand for essential oils, which has resulted in cases of adulteration. Authentication is thus a matter of critical importance for both consumers and chemical companies. This comprehensive overview covers known adulterations in essential oils, and some analytical methodologies adopted for their detection. We first list recommended tests, and then we explain and discuss common analytical techniques, such as chiral gas chromatography, isotope-ratio mass spectrometry, and nuclear magnetic resonance spectroscopy. We also present (high-performance) thin-layer chromatography, vibrational spectroscopy, coupled and multidimensional chromatography, high-performance liquid chromatography, and combination with chemometrics-metabolomics. This review provides a critical overview of existing techniques.
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Gas chromatographic retention indices were evaluated for 505 frequently reported plant essential oil components using a large retention index database. Retention data are presented for three types of commonly used stationary phases: dimethyl silicone (nonpolar), dimethyl silicone with 5% phenyl groups (slightly polar), and polyethylene glycol (polar) stationary phases. The evaluations are based on the treatment of multiple measurements with the number of data records ranging from about 5 to 800 per compound. Data analysis was limited to temperature programmed conditions. The data reported include the average and median values of retention index with standard deviations and confidence intervals.
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Thiabendazole, classified as antiparasitic and also used as an antifungal drug, can be found as otological solution indicated for treatment of parasitic and fungal external otitis in small animals. Malassezia pachydermatis is a yeast recognized as a normal inhabitant on the skin and mucous membranes of dogs and cats. However, it is considered an opportunistic agent that causes external otitis and dermatitis in these animals. The aim of this study was to evaluate the in vitro effect of thiabendazole against 51 isolates of M. pachydermatis using the CLSI Broth Microdilution method that has been adapted for this yeast species (NCCLS, 2002). Based on this test, the Minimum Inhibitory Concentrations (MIC) of thiabendazol was calculated. Subsequently, the susceptibility of each isolate against this antifungal was determined. It was observed that the MIC of thiabendazole against M. pachydermatis ranged from 0.03 to > 4 µg/mL. A total of 13.7% of the isolates were found to be resistant, 47.1% were intermediate and 39.2% were sensitive to the drug. The rate of resistance of the yeasts against thiabendazole was similar to the results previously obtained with other antifungals, while the adapted broth microdilution technique used in this study proved to be efficient.
In order to assess the kinetics of extraction Citrus aurantium leaves underwent hydrodistillation with a Clevenger apparatus. Essential oil was collected every 15 minutes during the whole extraction period. Yield variation during extraction period revealed an exponential aspect curve with 95% of maximum yield at 150 minutes. Sixty one compounds were identified during extraction time. Chromatographic analysis showed notable differences between the different petitgrain samples especially for the major components: linalool (29.74-62.43%), α-terpineol (3.04-15.69%) and linalyl acetate (5.64-25.38%). Important variations were observed in petitgrain chemical composition according to extraction time and each component presented a specific kinetic according to extraction time. These variations may be the expression of variable chemical and physical phenomena that take place in Clevenger hydrodistillation batch.
Citrus aurantium L. essential oil is commonly used as a flavouring agent. In the present study, the essential oil of fresh Citrus aurantium L. (CaEO) flowers cultivated in North East of Tunisia (Nabeul) was analyzed by GC-FID and GC-MS. 33 compounds were identified, representing 99% of the total oil. Limonene (27.5%) was the main component followed by E-nerolidol (17.5%), α-terpineol (14%), α-terpinyl acetate (11.7%) and E. E-farnesol (8%). The antimicrobial activity of the CaEO was evaluated against a panel of 13 bacteria and 8 fungal strains using agar diffusion and broth microdilution methods. Results have shown that the CaEO exhibited moderate to strong antimicrobial activity against the tested species. The investigation of the mode of action of the CaEO by the time-kill curve showed a drastic bactericidal effect after 5 min using a concentration of 624 μg/ml. The antioxidant activities of the CaEO were assayed by DPPH and beta carotene tests. Results showed that CaEO displayed an excellent DPPH scavenging ability with an IC50 of 1.8 μg/ml and a strong Beta-carotene bleaching inhibition after 120 min of incubation with an IC50 of 15.3 μg/ml. The results suggested that the CaEO possesses antimicrobial and antioxidant properties, and is therefore a potential source of active ingredients for food and pharmaceutical industry.
Five clones of sour orange (Citrus aurantium L.) showing significant morphological differences were selected from our germplasm collection and characterized both by genetic and leaf volatiles analysis. The genetic studies were undertaken by the use of molecular markers developed by PCR-based techniques (ISSR and RAPD), while the leaf essential oil patterns were obtained by chromatographic and mass spectrometric determination. Data obtained suggest that reasonably similar information can be achieved from the two techniques, supporting each other in characterization studies.