Artemisia dracunculus Essential Oil Chemical composition and antioxidant properties

Abstract

Free radical scavenging activity, total phenolic content and the chemical composition of the essential oil isolated by steam distillation from Artemisia dracunculus L. was investigated. The isolation yield was 0.24% (v/w) based on the fresh plant material (leaves). GC-MS investigation identified 21 components, accounting 99.93% of the total amount. The major components were sabinene (42.38%), isoelemicin (12.91%), methyl eugenol (9.09%), elemicin (7.95%) and beta-ocimene (6.46%). The free radical scavenging activity of the essential oil of Artemisia dracunculus L. was evaluated in vitro by the DPPH assay (IC50 = 0.730 ± 0.213 mg/mL), BHA and alpha-tocopherol were used as a positive control. The total phenolic content of the tarragon essential oil was evaluated by the Folin-Ciocalteu method (GAE = 0.451 ± 0.001 mg/g sample). In view of these data, we consider that tarragon essential oil could represent a new antioxidants source as a reliable option to reduce the usage of synthetic additives.

Full text

REV.CHIM.(Bucharest)♦70♦No. 12 ♦2019 http://www.revistadechimie.ro 59
Artemisia dracunculus
Essential Oil
Chemical composition and antioxidant properties
ARIADNA PETRONELA FILDAN1,2, IOAN PET3, DANIELA STOIN4, GABRIEL BUJANCA4,
ALEXANDRA TEODORA LUKINICH-GRUIA5, CALIN JIANU4*, ADELINA MARIA JIANU6*, MATILDA RADULESCU6,
DOINA ECATERINA TOFOLEAN1,7
1Ovidius University, Faculty of Medicine, 124 Mamaia Blvd., 900527, Constana, Romania
2Constanta Clinical Pulmonology Hospital, 167 Alexandru Lapusneanu Blvd., 900001, Constanta, Romania
3Banat’s University of Agricultural Sciences and Veterinary Medicine King Michael I of Romania from Timisoara, Faculty of
Bioengineering of Animal Resources, 119 Calea Aradului, 300645, Timisoara, Romania
4Banat’s University of Agricultural Sciences and Veterinary Medicine King Michael I of Romania from Timisoara, Faculty of Food
Engineering,119 Calea Aradului, 300645, Timisoara, Romania
5OncoGen Centre, County Hospital Pius Branzeu, 156 Liviu Rebreanu Blvd., 300736, Timisoara, Romania
6University of Medicine and Pharmacy Victor Babes, Faculty of Medicine, 2nd Eftimie Murgu Sq., 300041 Timisoara, Romania
7County Clinical Emergency Hospital of Constanta, 145 Tomis Blvd., 900591, Constanta, Romania
Free radical scavenging activity, total phenolic content and the chemical composition of the essential oil
isolated by steam distillation from Artemisia dracunculus L. was investigated. The isolation yield was 0.24%
(v/w) based on the fresh plant material (leaves). GC-MS investigation identified 21 components, accounting
99.93% of the total amount. The major components were sabinene (42.38%), isoelemicin (12.91%), methyl
eugenol (9.09%), elemicin (7.95%) and beta-ocimene (6.46%). The free radical scavenging activity of the
essential oil of Artemisia dracunculus L. was evaluated in vitro by the DPPH assay (IC50 = 0.730 ± 0.213
mg/mL), BHA and alpha-tocopherol were used as a positive control. The total phenolic content of the
tarragon essential oil was evaluated by the Folin-Ciocalteu method (GAE = 0.451 ± 0.001 mg/g sample). In
view of these data, we consider that tarragon essential oil could represent a new antioxidants source as a
reliable option to reduce the usage of synthetic additives.
Keywords: tarragon, essential oil, free radical scavenging activity, total phenolic content, antioxidant activity
* email: calin.jianu@gmail.com ; adelina.jianu@gmail.com
The genus Artemisia part of the family
Asteraceae
(Compositae), consisting over 500 diverse species
distributed principally in Europe, Asia and North America
[1].
Artemisia dracunculus
L. distribution spans over
western North America, eastern and central Europe, and
most of temperate Asia [2]. Tarragon (
A. dracunculus
) is a
medicinal plant known and used in folk medicine from
ancient times with anti-inflammatory, carminative,
antiparasitic, digestive, antispasmodic, antiseptic,
antipyretic and anthelmintic effects [1-3]. Also, tarragon
has been used for flavoring foods. His pleasant, spicy aroma
of the entire plant and its essential oil, represent the main
reasons for the extensive use in the food industry [2, 3].
Tarragon essential oil is obtained from aerial parts of the
plant during the flowering period [4]. The EO is a clear
liquid, pale yellow to amber in color, with a delicate spicy
odor; the extraction yield range between 0.15-3.1% [4-6].
Different studies have been carried out regarding
phytochemical and biological activity of the tarragon EO.
The main components of are methyl ethers, ocimene,
myrcene, α-pinene, β-pinene, camphene, limonene, and
linalool [1, 3, 5-7].
Biological effects including antibacterial, antifungal and
antioxidant activities have been previously reported for
A.
dracunculus
. Tarragon EO demonstrated antibacterial
activities over a vast spectrum, including human pathogens
such as
Pseudomonas aeruginosa
,
Escherichia coli
,
Staphylococcus aureus
,
Salmonella typhimurium
and
Staphylococcus epidermidis
[8-10]. Also, the
A.
dracunculus
EO has shown antifungal activity against some
fungal species including
Candida albicans
,
Cryptococcus
neoformans
,
Aspergillus niger
,
Microsporum canis
,
Trichophyton rubrum
,
Microsporum gypseum
and
Fonsecaea pedrosol
[8-12]. Additionally,
A. dracunculus
EO and some of his components demonstrate, in vitro, a
moderate radical scavenging activity [9-11].
Our study aimed to determine: i) the chemical
composition and ii) radical scavenging activity and total
phenolic content of the EO isolated from
A. dracunculus
leaves on which there are no previous studies.
Experimental part
Materials and methods
Reagents
2,2-diphenyl-1-picrylhydrazyl (DPPH); Folin Ciocalteu2
s phenol reagent (2N); anhydrous sodium sulfate; sodium
carbonate; methanol; hexane (Sigma-Aldrich, Germany);
butylated hydroxyanisole (BHA); alpha-tocopherol (Fluka
Analytical).
Raw material
Plant material of
A. dracunculus
(Russian variety) was
collected through the flowering period, in Ludestii de Jos,
Hunedoara County (Coordinates: 45°43’ 5” N 23°10’21”E
in August 2018. After the identification, a voucher specimen
was deposited in the Herbarium of the Faculty of Agronomy,
Banat’s University of Agricultural Sciences and Veterinary
Medicine King Michael I of Romania from Timisoara. Only
leaves, fresh material, was studied. The essential oil of
A.
dracunculus
was isolated by steam distillation, according
to the method previously described by Jianu et al. [13]. The
essential oil was separated by decantation, dried over
anhydrous sodium sulfate and stored (-18°C) in sealed
amber vials.
GC-MS analysis
The essential oil sample was diluted 1:1000 in hexane
before GC-MS injection and analyzed on an HP6890 Gas-

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Chromatograph coupled with HP5973 Mass Spectrometer.
One µL of sample was injected in the splitless mode on a
capillary column, Br-5MS, 5% Phenyl-arylene-95%
Dimethylpolysiloxane, 30 m length, 0.25 mm internal
diameter, 0.25 µm film thickness (Bruker). GC oven
temperature programme run in a range of 50°C to 300°C
with 6°C/min, with 5 min last hold and a solvent delay of 3
min. MS source was set at 230°C, MS Quad was set at
150°C, and ionization energy was 70 eV. The gas flow which
leads the sample through the column was helium at a
flow rate of 1 mL/min. The weight range of scanned
compounds was between 50 to 550 amu. All compounds
found were evaluated based on their spectra compared to
the mass spectrum from NIST0.2 library (USA National
Institute of Science and Technology software), area percent
was established. A semiquantitation based on the retention
times by calculating Kovats indexes and a comparison to
the Adams Indexes [14] from literature data was made.
Free Radical Scavenging Activity
The essential oil was analyzed regarding free radical
scavenging activity by a Brand-Williams’ adapted method
[15]. 0.1 mL sample methanolic solution of the essential
oil at different concentrations ranging from 1.5 mg/mL to
0.93 µg/mL/mL was placed in Corning 96 Flat Bottom clear
Polystyrol well plates. 0.1 mL methanol was used as the
control of DPPH. All samples were diluted in a ratio of 1:10
(v/v) DPPH/samples and incubated at room temperature
for 30 min , in darkness. The absorbance was measured at
515 nm with a Tecan i-control, 1.10.4.0 infinite 200Pro
spectrophotometer. The antioxidant activity was estimated
by the inhibition percent of the DPPH free radical and
calculated after the following formula:
I% = (Ablank – Asample/Ablank) • 100
where: Ablank express the absorbance of the control, and
Asample express the absorbance of the test sample.
IC50 was obtained using the BioDataFit 1.02 software
(Chang Broscience Inc, Castro Valley, CA, USA). Each test
was performed in triplicate.
Determination of Total Phenolics
The amount of total phenolic content was determined
according to the Folin-Ciocalteu method, adapted from
Swain and Hillis [16]. 15 mg sample was weighted and
dissolved in 1 mL methanol. A ratio of 1:5 sample and
Folin-Ciocalteu reagent (diluted 1:10 in distilled water) was
made and left for 5 min in the dark at room temperature;
after this reaction an equal volume with Folin-Ciocalteu
reagent of 7.5% sodium carbonate solution was added,
vortexed, and left one hour at room temperature, in
darkness. The absorbance was measured at 725 nm with
a spectrophotometer Tecan i-control, 1.10.4.0 infinite
200Pro spectrophotometer. The total phenolic content was
expressed in gallic acid equivalents (mg GAE/g sample)
after a propyl gallate calibration curve was made with
concentrations ranging between 0.375 mg/mL to 0.732
µg/mL.
Results and discussions
Hydrodistillation of
A. dracunculus
yielded 0.24% (v/w)
of essential oil, based on the fresh plant material. Previously
Arabhosseini et al. [17] report an amount of 0.6% essential
oil in the fresh leaves of
A. dracunculus
(a Russian variety).
Twenty-one volatile compounds were identified by their
mass spectra characteristics and retention indices using a
Br-5MS capillary column. The percentage of each
compound and their retention indices were presented in
table 1. The major compounds were sabinene (42.38%),
isoelemicin (12.91%), methyl eugenol (9.09%), elemicin
(7.95%) and beta-ocimene (6.46%). The EO is also
contained (Z)-beta-ocimene (4.78%) and terpinolene
(3.28%).
Arabhosseini et al. [17] investigating the chemical
composition of essential oil from Russian tarragon leaves
cultivated in the Netherlands report the presences of
sabinene (39.4%), methyl eugenol (14.7%), elemicin (16
%), isoelemicin (7.7%), (Z)-beta-ocimene (4.1%) and (E)-
beta-ocimene (3.1%) as the main components. Werker et
al. [18] report that essential oil from Russian tarragon
leaves mainly contained elemicin (36.2%), sabinene
(33.0%) and methyl eugenol (7.1%). These differences on
the chemical compositions of the
A. dracunculus
essential
oil can be partially attributable to environmental conditions
[2, 17, 18] and other factors such as genotypes and
ontogeny [2, 5, 19].
The free radical scavenging activity of the essential oil
of
A. dracunculus
was evaluated in vitro by the DPPH assay
(table 2). The tarragon essential oil demonstrated a low
radical scavenging activity compared with BHA and alpha-
tocopherol the positive controls used (Table 2). Our results
comply with previous studies that also reported a weak
DPPH radical scavenging activities for the
A. dracunculus
essential oils [9, 10]. In contrast, the
A. dracunculus
extracts evaluated in previous studies demonstrate a higher
free radical scavenging activity [20, 21]. Natural extracts,
including essential oils, are very complex mixtures of many
different compounds with distinct polarity as well as
antioxidant and prooxidant properties, sometimes showing
Table 1
THE CHEMICAL COMPOSITION OF THE ESSENTIAL OIL EXTRACTED
FROM LEAVES OF
A. dracunculus
CULTIVATED IN WESTERN ROMANIA

REV.CHIM.(Bucharest)♦70♦No. 12 ♦2019 http://www.revistadechimie.ro 61
synergic actions by comparison with individual compounds
[22]. Previously reports have demonstrated a linear
correlation between total phenolic content and antioxidant
activity in plants [20, 23-25]. This assumption is in accord
with our results, the essential oil analyzed by us showed a
low total phenol content (0.451 mg gallic acid / g sample).
However, numerous compounds of different polarity,
present in small amounts in the essential oils composition,
can also contribute to the antioxidant activity. Hydrolysis
and other separating methods are responsible for
separating this compounds from the plant material.
Additionally, the separating conditions (heat, extraction
systems, etc.) can modify the biological properties of these
natural extracts consisting of various compounds with
different chemical and physical properties [26]. Future
investigations are required to clarify the association
between antioxidant activity and chemical composition
of essential oils.
Conclusions
Summarizing the results, we conclude that the essential
oil isolated by steam distillation from
A. dracunculus
L.
leaves are rich in sabinene (42.38%), isoelemicin (12.91%),
methyl eugenol (9.09%), elemicin (7.95%) and beta-
ocimene (6.46%). Also, the tarragon essential oil
demonstrate antioxidant activity; besides, his scavenging
activity is lower in comparison to BHA and alpha-tocopherol
the positive controls used. In view of these data, we
consider that tarragon essential oil could represent a new
antioxidants source as a reliable option to reduce the usage
of synthetic additives.
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Table 2
YIELD, TOTAL PHENOLIC CONTENT AND DPPH RADICAL SCAVENGING ACTIVITIES OF THE ESSENTIAL OIL EXTRACTED
FROM LEAVES OF
A. dracunculus
CULTIVATED IN WESTERN ROMANIA
.

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Manuscript received: 15.09.2018