Arabian Journal of Chemistry (2020) 13, 8209–8217

King Saud University

Arabian Journal of Chemistry

www.ksu.edu.sa
www.sciencedirect.com

ORIGINAL ARTICLE

Chemical composition and antimicrobial activity of

the essential oils of Artemisia absinthium,
Artemisia scoparia, and Artemisia sieberi grown in
Saudi Arabia
Hanan Y. Aati a,*, Shagufta Perveen a,1, Raha Orfali a, Areej M. Al-Taweel a,
Sultan Aati b, Juergen Wanner c, Afsar Khan d, Rashad Mehmood e

a
Department of Pharmacognosy, College of Pharmacy, King Saud University. P. O. Box 22452, Riyadh 11495, Saudi Arabia
b
UWA Dental School, University of Western Australia, Nedland, Western Australia, Australia
c
Kurt Kitzing Co., Hinterm Alten Schloss 21, D-86757 Wallerstein, Germany
d
Department of Chemistry, COMSATS University Islamabad, Abbottabad Campus, Abbottabad 22060, Pakistan
e
Department of Chemistry, Division of Sience and Technology, University of Education, Township, Lahore, Pakistan

Received 3 August 2020; accepted 27 September 2020

Available online 8 October 2020

KEYWORDS Abstract Essential oils (EOs) are one of the most significant products of plant metabolites, the cur-
Essential oils; rent research work was performed to determine and compare the chemical compositions of the EOs
Antimicrobial activity; extracted from three different species of Artemisia (absinthium, sieberi, and scoparia) growing in
A. absinthium; Saudi Arabia and to test their antimicrobial potential against different bacterial and fungal strains.
A. scoparia; The EOs were isolated by hydro-distillation and analyzed by combining a gas chromatography–
A. sieberi; flame ionization detector (GC–FID) with the gas chromatography–mass spectroscopy (GC–MS)
GC–FID technique. Chemical analysis revealed that the three species had four compounds in common,
i.e., limonene, camphor, terpinen-4-ol, and ethyl 2-methylbutyrate, while the main components
identified in the EOs of A. absinthium and A. sieberi were cis-davanone (34.7% and 36.1%, respec-
tively) and camphor (16.2% and 24.1%, respectively). In contrast, the keto compounds dominated
in the oil of A. scoparia with 2-nonanone (55%) and 2-undecanone (24.5%) representing more than
80% of the total oil content. In addition, the antimicrobial activity of the isolated oils was evaluated
by the broth microdilution method, revealing that all the EOs isolated from the examined Artemisia
species displayed growth inhibiting actions in a concentration-dependent manner on selected tested

* Corresponding author.
E-mail addresses: [email protected] (H.Y. Aati), [email protected] (S. Perveen).
1
Co-corresponding author.
Peer review under responsibility of King Saud University.

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https://doi.org/10.1016/j.arabjc.2020.09.055
1878-5352 Ó 2020 The Author(s). Published by Elsevier B.V. on behalf of King Saud University.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
8210 H.Y. Aati et al.

microorganism species. The findings of the study also suggested that the tested EOs could be used to
develop effective natural antimicrobial remedies with potential application in the fields of cosmetic
industry, food manufacturing and medicine.
Ó 2020 The Author(s). Published by Elsevier B.V. on behalf of King Saud University. This is an open
access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction et al., 2010). Therefore, an additional aim of this study was

to identify the chemical diversity of three different species from
Medicinal plants have been widely used by humans for cen- the genus Artemisia, i.e., A. absinthium, A. sieberi, and A. sco-
turies as part of traditional medicine to treat various diseases paria. Considering also that in recent decades, several antimi-
and infections. To date, many studies have focused on collect- crobial herbal products have received considerable attention
ing information about the chemical content and activities of due to the rapid increase in the resistance of microorganisms
medicinal herbs, indicating that aromatic plants are important to antibiotics, we tested the activity of the obtained EOs
sources of secondary metabolites. Secondary metabolism in against Gram-positive and negative bacterial strains and a
plants not only contributes to their survival, but also acts as strain of filamentous fungi. The results of the study could be
a chemical defense against predators and diseases (Pandey therefore used to develop of novel antimicrobial drugs isolated
and Tripathi, 2011). The family Asteraceae is particularly rich from a natural source or contributing in food manufacturing
in plants that produce essential oils (EOs), among which the not only as flavorings but used as an interesting safe source
genus Artemisia holds a top position owing to its biological of natural antimicrobials for food preservation.
activities. This genus consists of small herbs and shrubs and
2. Materials and methods
comprises of about 500 species found in America, Asia, and
several European countries (Abad et al., 2012).
The Kingdom of Saudi Arabia is a vast arid land with an 2.1. Plant material
area of about 2,250,000 km2 that covers most of the Arabian
Peninsula, which is characterized by a wide diversity of plant The fresh aerial parts (each 500 g) of A. absinthium were col-
species and different ecosystems (Hanan et al., 2019). The lected from the city of Jazan and those of A. sieberi, and A.
genus Artemisia is represented in Saudi Arabia by seven species scoparia from the city of Riyadh, Saudi Arabia, in March
(A. scoparia, A. monosperma, A. sieberi, A. judaica, A. abyssi- 2018 during the flowering stage of plants. The plants were
nica, A. herba-alba, and A. absinthium) and has been used identified by the plant taxonomist Dr. Rajakrishnan Rajago-
worldwide in folk medicine since ancient times (Erel et al., pal, a botanist in the Science College Herbarium, King Saud
2012). The aromatic leaves of some Artemisia species are used University, Riyadh, Saudi Arabia.
for flavoring (as a culinary herbs). An in-depth literature sur-
vey revealed that different Artemisia species have been tradi- 2.2. Essential oils extraction
tionally used as antidiabetic, anthelmintic, tonic,
antimalarial, and antiulcer agents, as well as for the treatment The freshly cut aerial parts of A. absinthium, A. sieberi, and A.
of bronchitis, wounds inflammation, and tuberculosis. Many scoparia were subjected to hydro-distillation (HD) for 4 h
researchers have also explored the antioxidant, antipyretic, using a Clevenger apparatus and following the standard proce-
analgesic, antimicrobial, cytotoxic, and antifungal activities dure described in the literature (European Pharmacopoeia,
of different Artemisia species (Wright, 2002; Humara et al., 2005). The recovered oils were dissolved in petroleum ether
2014). The phytochemical studies confirmed that all classes and then filtered over anhydrous sodium sulfate to give deep
of activity principles exist in this genus with particular refer- blue (A. absinthium), blue (for A. sieberi), and colorless Eos
ence to terpenoids and flavonoids. Moreover, it has been (for A. scoparia) in yields of 0.83% (v/w), 0.42% (v/w), and
shown that the accumulation of EOs rich in sesquiterpene lac- 0.34% (v/w), respectively. Finally, the obtained oils were
tones and terpenoids results in strong aromatic odors and bit- stored in amber glass under N2 at 4 °C in the dark until their
ter tastes for most of the species, which may discourage further testing and analysis. The isolated EOs were diluted in
herbivores and lead to a selective advantage (Bora and dichloromethane (1:20) and 1 lL of this solution was injected
Sharma, 2011). An attractive application of essential oils and in the GC at a split ratio of 1:100.
their constituents is in food products to prolong the shelf-life
of foods by limiting growth or survival of microorganisms. 2.3. GC–FID and GC–MS analysis
To the best of our knowledge, the aroma volatiles of A.
absinthium grown in Saudi Arabia have not been explored in Gas chromatography–flame ionization detector (GC–FID)
detail so far. Thus, herein, we performed a detailed gas chro- and GC–MS analyses were performed in one run using a gas
matography–mass spectrometric (GC–MS) analysis for the chromatograph with a MS-FID-splitter consisting of a quartz
EOs isolated from the aerial part of wild grown A. absinthium Y-splitter, a short (ca. 20 cm) 0.1 mm id fused silica restrictor
in the southwestern region of Saudi Arabia. Abou El-Hamd column as an inlet to the GC–MS interface, and a ca. 1 m  0.
et al. (2010) has also confirmed that the various oil composi- 25 mm deactivated fused silica column serving as a transfer
tions observed in plants grown in different areas of the same line to the FID detector. The restrictor column was used to
country or different countries indicate the presence of several limit the flow in the MS vacuum and prevent the insertion of
oil-dependent chemotypes in the same plant (Abou El-Hamd combustion gases from the FID, which operated at atmo-
Chemical composition and antimicrobial activity of the essential oils 8211

spheric pressure. Moreover, the flow in the analytical column The MIC value for each strain was determined by the broth
had to be greater than the inflow to the MS detector, which microdilution method following a previous reported procedure
was limited to about 1 mL/min by the restriction line. The (Murthy et al., 2006). Each sample solution (10 mL) was loaded
GC column flow should also be constant, otherwise the FID/ on wells and dissolved in 10% dimethyl sulfoxide (DMSO).
MS split ratio would change with temperature. This configura- The final concentration of the oil was 2500 mg/mL. To achieve
tion yielded an FID and MS chromatogram with almost iden- a concentration range of 2500–5 mg/mL, a series of two-fold
tical retention times (RT), thus facilitating the assignment of dilutions of the stock solution were prepared. Erythromycin
the FID peaks to each substance. For these measurements, (Saudi Pharmaceutical Industry, SPI), amikacin (Bristol May-
we used a Thermo Fisher Scientific Trace GC Ultra with a ers Squibb, BMS), and itraconazole (SAJA) were used as the
split/split less injector heated at 250 °C and connected to a positive reference standards for the Gram-positive bacteria,
50 m  0.25 mm  1.0 mm SE-52 (95% polydimethylsiloxane, Gram-negative bacteria, and fungi, respectively. Two-fold
5% polydiphenylsiloxane) capillary column (prepared and dilutions of the positive controls with concentrations of
tested for deactivation and separation efficiency in our lab 1000–1 mg/mL were also prepared. The respective bacterial
(Kurt, 1986), an FID detector operating at 250 °C, and a Tri- and fungal inoculums were of 104 CFU/mL and 103 CFU/
Plus RSH Autosampler. mL, respectively, which were incubated in 96-well microplates
For the identification of the EO components, a Thermo at 37 °C for 24 and 48 h for bacteria and fungi, respectively.
Fisher Scientific ISQ mass spectrometer was used with a GC/ The experiments were performed in duplicate and repeated
MS interface heating at 250 °C, an ion source operating at three times (n = 3). Accordingly, the lowest concentration
230 °C, an electron ionization mode at 70 eV, a filament with of the tested oil that could inhibit the bacterial growth was
50 mA, and a scan range of 40–500 amu. The oven temperature expressed as MIC in lg/mL (Joshi, 2010).
program involved heating at 60 °C for 1 min, then heating to
230 °C at a rate of 3 °C/min, and a 230 °C isotherm for 2.6. Statistical analysis
12.3 min. The carrier gas was helium 5.0 with a constant flow
rate of 1.5 mL/min. All data were expressed as mean ± standard deviation (SD)
values of three replicates. Where applicable, the data were sub-
2.4. Identification of the essential oil components jected to one-way analysis of variance. Based on the statistical
package analysis of Microsoft Excel 2010, the differences were
The Thermo Xcalibur 2.2 software was used to identify the EO considered statistically significant for p values of <0.05.
compounds by correlating the obtained mass spectra with the
databases of the National Institute of Standards and Technol- 3. Results
ogy (NIST 08), Wiley 8th ed. (Wiley RegistryTM), Adams
library (Adams, 2007), Mass Finder terpenoids library The application of HD to the aerial parts of the three Artemi-
(König et al., 2001), and our own library. However, the simple sia species (A. absinthium, A. sieberi, and A. scoparia) provided
comparison of the mass spectra is not sufficient for a clear deep blue, blue, and colorless EOs in yields of 0.83% (v/w),
identification, especially for sesquiterpenes. Thus, the position 0.42% (v/w), and 0.34% (v/w), respectively, depending on
in the chromatogram was also considered as a second criterion the fresh weight. The complexity of the chemical composition
determined by comparing the calculated retention indices (RI) of the EOs isolated from the aerial parts of A. absinthium, A.
of the peaks with the corresponding literature data, NIST 08, sieberi, and A. scoparia, as determined by GC–MS analysis,
Adams (2007) and König et al. (2001) or reference compounds. is demonstrated in Table 1.
The RI values were determined by measuring the RTs of a ser- The volatile constituents of the three EOs were identified
ies of n-alkanes that were eluted across the entire chro- based on the MS libraries (NIST 08, Wiley 8th ed., Adams
matogram and were calculated according to the method of library, Mass Finder terpenoids library) and by comparing
Van den Dool and Kratz (Van den Dool and Kratz, 1963; the obtained data with the corresponding MS literature find-
Gas Chromatographic Retention Data, NIST). Moreover, ings. The relative percentages of the individual components
the EO components were quantified using normalized peak were calculated based on the GC peak areas. Hence, GC–
area calculations of the FID chromatogram without (by first MS analysis revealed the presence of 57 constituents in the
approximation) relative FID-response factors. A. absinthium oil, which represented 95.1% of the total oil.
The major constituents of this oil were cis-davanone
2.5. Antimicrobial assay (34.7%), camphor (16.2%), chamazulene (8.2%), E-nerolidol
(4%), and terpinen-4-ol (3.4%). The A. sieberi oil consisted
The antimicrobial activity of the isolated EOs was evaluated of 54 volatile components, which represented 97.8% of the
by calculating their minimum inhibitory concentration (MIC) total oil content, while the main volatile compounds were
against Gram-positive [Staphylococcus aureus (CP011526.1), cis-davanone (36.1%), camphor (24.1%), terpinen-4-ol
Bacillus licheniformis (KX785171.1), and Micrococcus luteus (5.4%), E-nerolidol (5%), linalool (3.6%), bornyl acetate
(NCTC 2665)] and Gram-negative [Enterobacter xiangfangen- (3.6%), and chamazulene (2.9%). Furthermore, the GC anal-
sis (CP017183.1), Escherichia fergusonii (CU928158.2), and ysis of the A. scoparia oil indicated the presence of 23 aroma
Pseudomonas aeruginosa (NR-117678.1)] bacterial strains. In constituents, among which 2-nonanone (55%) and 2-
addition, their activity against three pathogenic fungi, i.e., undecanone (24.5%) represented more than 80% of the total
Candida albicans (MF942350), Candida parapsilosis oil content. A comparison of the findings for the three plant
(MF942354), and Aspergillus parasiticus (CBS 100926), was species indicated that the oxygenated sesquiterpenes and
examined. monoterpenes were the major chemical classes in the EOs
8212 H.Y. Aati et al.

Table 1 Volatile compounds of the essential oils isolated from the A. absinthium, A. sieberi, and A. scoparia aerial parts and their RTs
and RIs, as determined by GS–MS analysis.
No. Compounds* RT (min) A. absinthium A. sieberi A. scoparia RI
1 Ethyl 2-methylbutyrate 10.83 1.0 1.1 tr. 844
2 Ethyl isovalerate 10.94 0.2 0.1 – 846
3 a-Thujene 14.67 0.2 0.2 – 930
4 a-Pinene 15.12 2.1 2.1 – 940
5 Propyl 2-methylbutyrate 15.20 0.6 1.0 – 941
6 Camphene 15.90 2.0 2.0 – 956
7 2-Octanone 17.37 – – 0.4 986
8 Myrcene 17.55 1.1 0.6 – 990
9 a-Terpinene 19.08 0.8 0.6 – 1021
10 p-Cymene 19.42 0.3 0.5 – 1028
11 Limonene 19.70 0.3 0.4 0.1 1034
12 c-Terpinene 21.14 1.4 1.2 – 1062
13 2-Nonanone 22.48 – 0.6 55.0 1088
14 Terpinolene 22.67 0.4 0.3 – 1093
15 2-Nonanol 22.84 – – 2.6 1097
16 Linalool 22.93 3.2 3.6 – 1098
17 Nonanal 23.07 – – 0.8 1101
18 trans-Sabinene hydrate 23.14 0.2 0.2 – 1103
19 cis-p-Menth-2-en-1-ol 24.34 0.1 0.2 – 1127
20 2-Octanol acetate 24.66 – – 0.6 1134
21 trans-p-Menth-2-en-1-ol 25.19 0.1 0.2 – 1145
22 Camphor 25.63 16.2 24.1 1.1 1154
23 Nonanol 26.31 – – 0.3 1168
24 Borneol 26.66 0.4 0.8 – 1175
25 Terpinen-4-ol 27.15 3.4 5.4 0.1 1185
26 2-Decanone 27.39 – – 1.7 1190
27 a-Terpineol 27.71 0.3 0.4 – 1196
28 2-Nonanol acetate 29.38 – – 9.3 1232
29 Bornyl acetate 32.26 2.0 3.6 0.0 1293
30 2-Undecanone 32.20 – – 24.5 1292
31 Nonyl acetate 32.50 – – 0.6 1298
32 Myrtanol acetate 33.01 0.4 0.4 – 1309
33 Ethyl hydrocinnamate 34.85 0.3 0.4 – 1351
34 6-Dodecanone 35.44 – – 0.5 1364
35 Z-Ethyl cinnamate 36.07 0.6 0.6 – 1378
36 Limonen-10-yl acetate 37.61 0.2 0.3 – 1413
37 2-Acetoxyundecane 38.22 – – 0.8 1428
38 E-b-Caryophyllene 38.77 0.3 0.1 – 1441
39 E-Ethyl cinnamate 39.98 2.5 1.6 – 1469
40 Davana ether isomer 1 40.96 0.3 – – 1493
41 b-Selinene 41.59 0.2 0.1 – 1508
42 Davana ether isomer 2 41.78 1.6 tr. – 1513
43 Davana ether isomer 3 42.53 0.6 – – 1532
44 Davanone B (allo-Davanone) 43.82 1.0 – – 1564
45 E-Nerolidol 43.97 4.0 5.0 – 1568
46 Davanone 44.08 1.8 – – 1570
47 iso-Davanone 44.34 0.6 – – 1577
48 cis-Davanone 45.01 34.7 36.1 – 1594
49 Davanol 46.01 0.3 0.2 – 1620
50 Methyl jasmonate 47.25 0.6 0.1 – 1652
51 Chamazulene 51.09 8.2 2.9 – 1756
Oxygenated monoterpenes 26.7 39.5 1.2 –
Monoterpene hydrocarbons 8.7 8.1 0.1 –
Oxygenated sesquiterpenes 44.9 41.3 – –
Sesquiterpene hydrocarbons 8.8 3.2 – –
Saturated fatty acids 1.9 2.3 0.4 –
Miscellaneous 4.1 3.4 96.9 –
Total (%) 95.1 97.8 98.6 –
Rt = Retention time (min.), RI = Retention indices.
*
The isolated compounds are listed following their order of elution from the HP-5 MS column.
Chemical composition and antimicrobial activity of the essential oils 8213

obtained from A. absinthium and A. sieberi, while the keto cated that the main chemical classes of components were oxy-
compounds dominated in the A. scoparia oil (Fig. 1). The genated sesquiterpenes (41.3%), which were mainly
GC–MS total ion chromatogram of the oils is shown in represented by davanone, its alcohol derivative, and camphor,
Figs. 2-4. and monoterpenes (39.5%), including terpinen-4-ol, linalool,
Traditionally, A. absinthium, A. sieberi and A. scoparia used and bornyl acetate (Fig. 3).
an anthelmintic, antiparasitic, antihypertensive, relieve ten- In contrast to the other two species, the composition of the
donitis, earache, bronchitis, wound burns infection and to cure A. scoparia EO was dominated by the ketones, 2-nonanone
different type of infections. For that, we assessed quantita- and 2-undecanone, Table 1 and Fig. 4 which represented more
tively the antimicrobial and antifungal activities of the three than 80% of the total oil content (98.6%).
tested EOs by determining their MIC values to validate the The chamazulene content in the A. absinthium and A. sie-
folkloric use for these plants. As shown in Table 2, the EOs beri species was 8.2% and 2.9%, respectively, giving the corre-
of A. sieberi and A. scoparia showed the most significant activ- sponding oils a blue color with varying intensity depending on
ities against both Gram-negative and -positive bacteria. Espe- the chamazulene percent concentrations (Rezaeinodehi and
cially the EO of A. scoparia exhibited the lowest MIC against Khangholi, 2008) (Figs. 2 and 3).
Staphylococcus aureus, Enterobacter xiangfangensis, and Can- Davanone is a sesquiterpene lactone found in various Arte-
dida albicans (0.2 ± 0.3, 3.0 ± 0.1 and 0.1 ± 0.5 lg/mL, misia species (Luigi et al., 2016) and has a chemotaxonomic
respectively) with higher inhibition values than the positive value. The davana (A. pallens) oil contains davanone as major
control broad-spectrum standard antimicrobial drugs ery- constituent and exhibits antispasmodic, antibacterial, antifun-
thromycin, amikacin, and itraconazole, 2.0 ± 1.0, 9.0 ± 4.0 gal properties, as well as wound healing, antihelmintic, hypo-
and 0.29 ± 0.6 lg/mL, respectively. Moreover, the EO iso- glycemic, and antidepressant activities also, it used as food
lated from A. sieberi was the most active against Micrococcus flavoring (Shreyas et al., 2018). As previously mentioned, the
luteus with a MIC value of 0.5 ± 0.1 lg/mL, whereas the EO EOs isolated from the A. absinthium and A. sieberi Saudi spe-
of A. absinthium was more sensitive to Gram-positive bacteria cies were dominated by cis-davanone (34.7% and 36.1%,
and fungi and completely inactive against the selected Gram- respectively). Thus, the present study could also set the basis
negative bacteria (Table 2, Fig. 5). for the isolation of davanone from a natural source that is
widely used in the medical field as well in food industry. Fur-
4. Discussion thermore, davanone has been reported to be odorless, there-
fore, the odoriferous constituents of oils containing
The A. absinthium oil was also characterized by a high content davanone were attributed to the presence of davana ethers
of oxygenated compounds including both sesquiterpenes (Thomas and Dubini, 1971). Herein, three out of four natural
(44.9%) and monoterpenes (26.7%) (Table 1, Fig. 1). More davana ether stereoisomers (Garneau et al., 2013) were
specifically, davanone and its derivatives were the most com- detected in the EO isolated from A. absinthium, while traces
mon sesquiterpenes, accounting for approximately 40.8% of from davana isomer 2 were identified in the A. sieberi EO. In
the total detected compounds, while camphor was the predom- contrast, no davana ethers were detected in the A. scoparia oil.
inant monoterpene with a concentration of 16.2% in a total A previous study on the leaf extract of A. sieberi growing in
monoterpene concentration of 26.7% (Fig. 2). Saudi Arabia led to the isolation of davanone (Lamya et al.,
Furthermore, the main compounds isolated from the A. sie- 2020), but there was no reference to the content of Saudi A.
beri distilled oil were cis-davanone, camphor, terpinen-4-ol, E- sieberi EOs in davanone. Instead, an Iranian report confirmed
nerolidol, linalool, bornyl acetate, and chamazulene, which that the EO isolated from A. sieberi contains davanone in 15%
accounted for approximately 97.8% of the total oil content v/w, which is, significantly lower that the value determined in
(Table 1). Moreover, the GC–MS and GC–FID analysis indi- this study for the Saudi species (36.1%v/w). Arbi et al. also

Fig. 1 Keto compounds classes identified in the essential oils of A. absinthium, A. sieberi, and A. scoparia.
8214 H.Y. Aati et al.

Fig. 2 GC–FID chromatograms of the volatile compounds in the A. absinthium essential oil. The numbers represent the peaks of (1)
ethyl isobutyrate, (2) ethyl 2-methylbutyrate, (3) a-thujene, (4) a-pinene, (5) camphene, (6) myrcene, (7) c-terpinene, (8) linalool, (9)
camphor, (10) terpinen-4-ol, (11) a-terpineol, (12) bornyl acetate, (13) Z-ethyl cinnamate, (14) E-ethyl cinnamate, (15) davana ether isomer
2, (16) davanone B (allo-davanone), (17) E-nerolidol, (18) davanone, (19) cis-davanone, (20) methyl jasmonate, and (21) chamazulene.

Fig. 3 GC–FID chromatograms of the volatile compounds in the essential oil obtained from A. sieberi. The numbers represent the peaks
of (1) ethyl 2-methylbutyrate, (2) a-pinene, (3) camphene, (4) c-terpinene, (5) linalool, (6) camphor, (7) terpinen-4-ol, (8) bornyl acetate, (9)
E-ethyl cinnamate, (10) E-nerolidol, (11) cis-davanone, and (12) chamazulene.

reported that the EO isolated from the A. sieberi Saudi species the currently investigated A. absinthium Saudi species was
collected from the northern region of the Kingdom was dom- 34.7%v/w out of the total oil content. Given that, to the best
inated by spathulenol (30.4%v/w) (Arbi et al., 2017). of our knowledge, no studies have so far explored the EO of
In addition to the A. sieberi species, the EO isolated from the A. absinthium Saudi species, our study can be considered
the A. absinthium Ethiopian species included 26.5% davanone the first report. Nevertheless, several studies on the EOs of dif-
(Asfaw and Demissew 2015), while the davanone content in ferent Artemisia species have attributed their antimicrobial
Chemical composition and antimicrobial activity of the essential oils 8215

Fig. 4 GC–FID chromatograms of the volatile compounds in the essential oil obtained from A. scoparia. The numbers represent the
peaks of (1) 2-nonanone, (2) 2-nonanol, (3) camphor, (4) 2-decanone, (5) 2-nonanol acetate, (6) 2-undecanone, and (7) 2-acetoxyundecane.

activity to their davanone-type sesquiterpenes content (Vlatka Table 1 showed some similar compounds could be detected
et al., 2004; Stefanie et al., 2008). in all three Artemisia species EOs (limonene, camphor, and
In contrast to the other two species, the EOs isolated from terpinen-4-ol), but their quantities were different. Although
the aerial part of A. scoparia that was collected from the cen- the plant aerial parts were collected from the same country
tral region of Saudi Arabia consisted mainly of 2-nonanone and belonged to the same family and genus, the Artemisia spe-
and 2-undecanone. However, the oil isolated from the same cies exhibited chemical polymorphism, which was attributed to
species growing in the same country (Saudi Arabia) but col- the chemovariation of EOs due to the different genetic and
lected from the northern region of Saudi Arabia was domi- environmental (temperature, moisture, and chemical composi-
nated by acenaphthene (83.2%v/w) (Arbi et al., 2017). tion of soil) factors that affected the EO compositions. There-
Previous studies have also shown that the insecticidal and fore, by comparing the current results with relevant data from
antimicrobial activities of EOs isolated from A. herba-alba previous studies we concluded that the geographic origin con-
are mainly due to their ketone content (Herrera et al., 2015; siderably affects the chemical composition of the Artemisia
Nassiba et al., 2017). Therefore, since A. scoparia contains species.
the highest percent concentration of ketones, we assumed that In recent years, the resistant isolates of microorganisms
the presence of ketone groups may increase the toxicity against have significantly increased (Bingyun and Thomas, 2018) due
microbes. to the increasing use of antibiotics. Taking also into account
Composition and yield of secondary metabolites in plants the adverse effects caused by chemical agents used for the
like EOs, can be affected in a number of ways, from their for- treatment of infectious diseases, the research community has
mation in the plant to their isolation. Several factors have been also focused on the discovery of new natural compounds. In
studied, in particular for commercially important crops, to this study, it was revealed that the examined Gram-positive
optimize the cultivation conditions and time of harvest and bacteria were more susceptible to the antibacterial activity of
to obtain higher yields of high-quality essential oils. In addi- EOs than the Gram-negative bacteria. This higher resistance
tion to the commercial importance of the variability in yield of the Gram-negative bacteria could be attributed to their
and composition, the possible changes are also important external membrane, which surrounds their cell wall and inhi-
when the essential oils and volatiles are used as chemotaxo- bits the penetration of the hydrophobic EOs through its
nomic tools. Knowledge of the factors that determine the lipopolysaccharide membrane that would affect the cell struc-
chemical variability and yield for each species are thus very ture and functionality. Additionally, the external capsule of
important. These include: physiological variations (organ some Gram-negative bacteria limits or prevents the diffusion
development, pollinator activity cycle, type of plant material of EOs into the bacterial cell (Ratledge and Wilkinson,
[leaves, flowers, etc.], type of secretory structure, seasonal vari- 1988). A previous study has demonstrated that the most abun-
ation and mechanical or chemical injuries); environmental dant chemical groups in the A. absinthium and A. sieberi oils,
conditions (climate, pollution, diseases and pests and edaphic i.e., oxygenated monoterpenes, such as 1,8-cineole, camphor,
factors); geographic variations; genetic factors and evolution; 1-terpinen-4-ol, linalool, a-terpineol, borneol, and bornyl acet-
storage; political/social conditions; and also amount of plant ate, have potent antimicrobial activities (Carson and Riley
material needs (Cristina et al., 2008). 1995). Moreover, the antibacterial and antifungal activities
8216 H.Y. Aati et al.

Table 2 MIC (mg/mL) values of the A. absinthium, A. sieberi, and A. scoparia essential oils and the corresponding values of three
broad-spectrum standard antimicrobial drugs.
Microbial strains MIC [mean ± SD (mg/mL)]*
A. absinthium A. sieberi A. scoparia Erythromycin Amikacin Itraconazole
Gram-positive
S. aureus CP011526 5 ± 0.7 2 ± 0.6 0.2 ± 0.3 2.0 ± 1.0 – –
B. licheniformis KX785171 11 ± 0.8 4 ± 0.3 2 ± 0.2 1.0 ± 1.0 – –
M. luteus NCTC2665 8 ± 0.4 0.5 ± 0.1 4 ± 0.1 1.0 ± 1.0 – –
Gram-negative
E. xiangfangensis CP017183 >25 16 ± 0.2 3 ± 0.1 – 9.0 ± 4.0 –
E. fergusonii CU928158 >25 12 ± 0.5 > 25 – 7.0 ± 8.0 –
P. aeruginosa NR117678 >25 17 ± 0.1 >25 – 4.0 ± 3.0 –
Fungi
C. albicans MF942350 4 ± 0.1 2 ± 0.0 0.1 ± 0.5 – – 0.29 ± 0.6
C. parapsilosis MF942354 5 ± 0.6 3 ± 0.2 2 ± 0.3 – – 0.33 ± 0.4
A. parasiticus CBS 100926 2.5 ± 0.5 1 ± 0.1 4 ± 0.1 – – 0.28 ± 1.3
*
MIC > 25 lg/mL.

Fig. 5 MIC (mg/mL) of the essential oils obtained from A. absinthium, A. sieberi, and A. scoparia.

of the oxygenated sesquiterpenes, which were the main con- different regions of Saudi Arabia. The results indicated
stituents of the A. absinthium and A. sieberi oils accounting that the chemotype of the studied Artemisia EOs was
for 44.9% and 41.3%, respectively, of the total EOs content specific and different from other already reported Artemi-
and consisted of a high amount of davanone and its deriva- sia EOs chemotypes. In addition, the tested Artemisia
tives, have also been established (Shreyas et al., 2018). In addi- EOs showed good antimicrobial activities, which indicated
tion, the antimicrobial properties of the hydrocarbons of their effectiveness against diseases caused by the overpro-
monoterpenes and sesquiterpenes that were also identified in duction of microorganisms. Thus, these species could be
high concentrations in these EOs have also been recognized further investigated to develop new antimicrobial agents,
(Adorjan and Buchbauer, 2010). Especially chamazulene has can be used as natural safe additives in food products
been identified as a fungicidal agent against 34 species of fungi to prolong the shelf-life of foods by limiting growth or
at a dose of 20 mL (Kordali et al., 2005). Minor components survival of microorganisms. However, additional studies
should be considered as well as, they may also play a critical are required regarding the safety and toxicity of the indi-
role in the EOs antibacterial activity through their synergistic vidual major compounds, as well as the effectiveness of
effect with other oil components (Lopes-Lutz et al., 2008). the EOs as a source of active biological compounds in
animal models.
5. Conclusions Funding

The present study investigated the chemical compositions This work was supported by the Deanship of Scientific
and tested the antimicrobial activity of three Artemisia Research at the King Saud University through the Research
species by isolating volatile EOs from plants growing in Group Project no RGP-221.
Chemical composition and antimicrobial activity of the essential oils 8217

Authors’ contributions König, W.A., Hochmuth, D.H., Joulain, D., 2001. Terpenoids and
Related Constituents of Essential Oils. Library of Mass Finder 2.1.
Institute of Organic Chemistry, Hamburg.
HYA and SP conceived and designed the experiments; HYA, Kordali, S., Kotan, R., Mavi, A., Cakir, A., Ala, A., Yildirim, A.,
SP, RO and JW performed the experiments; HYA and SP ana- 2005. Determination of the chemical composition and antioxidant
lyzed the data and wrote the primary draft of the manuscript; activity of the essential oil of Artemisia dracunculus and of the
SY participated in the sample collections and manuscript writ- antifungal and antibacterial activities of Turkish Artemisia
ing and editing; All authors have read and agreed to the pub- absinthium, A. dracunculus, Artemisia santonicum, and Artemisia
lished version of the manuscript. spicigera essential oils. J. Agric. Food Chem. 53, 9452–9458.
Kurt, G., 1986. Making and Manipulating Capillary Columns for Gas
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