Anal. Methods Environ. Chem. J. 5 (4) (2022) 66-76
Research Article, Issue 4
Analytical Methods in Environmental Chemi s try Journal
Journal home page: www.amecj.com/ir
AMECJ
Evaluating the eect of ethanol foliar feeding on the essential oil,
phenolic content, and antioxidant activities of Ducrosia anethifolia
Aliyeh Sarabandia, Amirhossein Sahebkarb,c,d,e, Javad Asilif, Moharam Valizadehg, Khalilollah Taherih,
Jafar Valizadeh,*, and Maryam Akaberif,*
a Department of Phytochemis try, Faculty of Science, University of Sis tan and Baluchis tan, Zahedan, Iran.
bApplied Biomedical Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
c Biotechnology Research Center, Pharmaceutical Technology Ins titute, Mashhad University of Medical Sciences, Mashhad, Iran.
d School of Medicine, The University of Wes tern Aus tralia, Perth, Wes tern Aus tralia, Aus tralia.
e Department of Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
f Department of Pharmacognosy, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
g Faculty of Environmental Sciences and Sus tainable Agriculture, Sis tan and Baluchis tan University, Zahedan, Iran.
h Department of Biology, Faculty of Science, University of Sis tan and Baluchis tan, Zahedan, Iran
AB S TRACT
Ducrosia anethifolia (Apiaceae) is a medicinal aromatic plant
di s tributed in Iran and Afghani s tan. This research aims to inve s tigate
the composition of the plant essential oil, determine the total avonoid
and phenolic contents, and evaluate its antioxidant activities after
ethanol foliar feeding. For this purpose, 0, 10, 20, 40, and 80%
v/v of aqueous ethanol solutions were sprayed on dierent batches
of the plants. Then, the essential oils were obtained using water
di s tillation. Compounds were analyzed by Gas chromatography-
mass spectrometry technique(GC-MS) using a validated method.
The method was validated as per the ICH guidelines for linearity,
precision, accuracy, robu s tness, LOD, and LOQ. The total contents
of phenols and avonoids were measured using spectrophotometric
methods. The antioxidant activity was evaluated using DPPH and
FRAP assays. The results showed that n-decanal, cis-verbenyl
acetate and dodecanal were the major compounds in all treatments.
However, alcohol could cause signicant dierences in the essential
oils qualitatively and quantitatively. The results showed that 40%
ethanol could increase the number of phenolics and avonoids and
consequently the antioxidant activity. Thus, ethanol foliar feeding
can be used as an appropriate approach to increase the essential oil of
D. anethifolia as well as its phenolic and avonoid contents.
Keywords:
Ducrosia anethifolia,
GC-MS,
Spectrophotometry,
Foliar feeding,
Essential oil,
Antioxidant
ARTICLE INFO:
Received 27 Jul 2022
Revised form 11 Oct 2022
Accepted 23 Nov 2022
Available online 28 Dec 2022
*Corresponding Author: Maryam Akaberi and Jafar Valizade
Email: akaberim@mums.ac.ir and djafar.walisade@gmail.com
https://doi.org/10.24200/amecj.v5.i04.213
------------------------
1. Introduction
Ducrosia anethifolia (DC.) Boiss. (A s teraceae) is
an aromatic herbaceous and biennial plant with
a height of 10–30 cm. The s tems are glabrous
and branched mo s tly from the base. The leaves
are ovate-oblong, 2–6 cm long, and branched,
with a petiole length of 5–18 cm. The edges of
the petals are jagged and slightly shaggy and the
compound umbel inorescence has white owers.
In Iran, the plant is known as Moshgak, Mushk
Boo, Darvishan Ginger, Reshgak, Khorkhundai,
Gavarshkh, and mount Coriander [1]. The genus
Ducrosia has three species in Iran including D.
anethifolia, D. assadii Alava, and D. abellifolia
Boiss. Ducrosia anethifolia grows wildly in
mountainous and plain areas on sandy soils in
67
dierent regions of Iran such as Kerman, Khorasan,
Zanjan, Shushtar, Behbahan, Shiraz, Kazerun,
Borazjan, Noorabad, Farashband, Firoozabad,
Jahrom, Darab, and Si s tan Baluche s tan [2]. It also
grows natively in countries from the Mediterranean
range to W. Paki s tan such as Afghani s tan, Paki s tan,
Iraq, Syria, Lebanon, and some Arab countries
[3]. Ducrosia assadii is endemic to Iran and D.
abellifolia is native to Syria, W. Iran, and Arabian
Peninsula [4]. Essential oils are one of the mo s t
pharmacologically important con s tituents in plants
belonging to Ducrosia. Terpene compounds are
reported to be responsible for many medicinal
activities of the oil of this medicinal plant. The
volatile compounds of this aromatic plant are
used as a avoring additive in various beverages
and desserts. Besides, Moshgak is also used as
an edible vegetable [5, 6]. Several s tudies have
inve s tigated the essential oil composition of D.
anethifolia revealing that the long chain oxygenated
hydrocarbons such as decanal, dodecanal, and
decanol con s titute the major compounds in the
essential oil. In addition, monoterpenes including
α-pinene, α-thujene, linalool, cis-citronellyl
acetate, and oxygenated sesquiterpenes such
as chrysanthenyl acetate have been reported as
the main con s tituents of the essential oil [7]. In
Iranian traditional medicine, the aerial parts of D.
anethifolia have been used to relieve various pains
such as headaches, back pain, and colic pain. It
has been also used for the treatment of seizures,
insomnia, heartburn, cataract, inammation of the
inner wall of the nose, and colds. Pharmacological
s tudies show that the plants belonging to this genus
have muscle relaxant, CNS depressant, and anti-
depressant properties. In addition, the essential oils
obtained from D. anethifolia have antimicrobial
properties again s t gram-positive bacteria, yea s ts,
and some dermatophytes. The essential oil of D.
anethifolia is reported to have antifungal properties
by preventing the growth of parasitic fungi such as
Candida albicans on the skin [8-10]. It could also
improve kidney function and lower the lipid levels
of the blood. Alpha-pinene as one of the major
compounds in essential oil is probably responsible
for the anti-anxiety eect of the plant. Myrcene,
as another main component of the plant, has
several pharmacological activities including anti-
radical, inhibitory eects, anti-cancer, and anti-
tumor properties [11-13]. The use of methanol and
ethanol foliar feeding is one of the mo s t important
approaches for increasing plant growth and harve s t
yield. Research has shown that ethanol becomes
acetaldehyde after penetration into the plant tissue.
Acetaldehyde is transformed into acetate (acetic
acid) by the acetaldehyde dehydrogenase enzyme.
Acetic acid also converts to acetyl coenzyme A,
which eventually turns into carbon dioxide and
dioxide. Methanol, ethanol, and other alcohols
are non-toxic to plants and can simply penetrate
the membrane of plant cells. The absorption
rate directly depends on the density of alcohol.
Therefore, the application of methanol and ethanol
foliar feeding on the aerial parts of C3 plants (the
plants that only use the s tandard method of carbon
dioxide xation by the enzyme Rubisco) [14], in
which their light breathing is large can compensate
for part of the s tabilized carbon losses and in this
way, increase pure photosynthesis and dry matter
production per unit area. As a result, some s tudies
conducted in the eld of agronomic C3 plants have
shown that methanol could aect the performance
of these plants positively [15]. Thus, the aim of
this s tudy was to evaluate the eect of ethanol
foliar feeding on the essential oil composition and
yields of D. anethifolia. In addition, the antioxidant
activities of the plant extract were inve s tigated and
the total phenol and avonoid contents of the plant
were measured.
2. Material and Methods
2.1. Planting and harves ting
This s tudy was performed at the research farm of
the research center for medicinal and ornamental
plants of Si s tan and Baluche s tan University with a
latitude of 29°27’N and a longitude of 60°51’E at an
altitude of 1410 m from sea level. In this experiment,
seeds of the plant samples were planted in 7 rows
of binary, each row was about 80 centimeters
and the di s tance between every two rows was 20
Determination and evaluation of the aromatic essential oil in plant Aliyeh Sarabandi et al
68
centimeters. The plants were divided into ve
treatment groups including control (di s tilled water)
and four concentrations of ethanol solution (10, 20,
40, and 80%). The ethanol spraying was performed
before the owering s tage of the plants every three
days 6 times. The spraying process began in early
May and the plants were harve s ted in early June
(spring 2016). After harve s ting, the aerial parts of
the plants were dried in the shade and s tored until
use. After collecting, the plants were transferred
to the laboratory and dried in the shade. Then, the
plants were milled, powdered, and prepared for
essential oil.
2.2. Extraction Procedure
The dried aerial parts of the plants for each treatment
were powdered separately with an electric mill.
Then, the powdered samples were subjected to
extraction. The essential oil of the samples was
obtained by water di s tillation. For this purpose,
50 g of each dried plant sample was subjected to
hydrodi s tillation for 3 h using a Clevenger-type
apparatus (1000 ml water). The essential oils were
collected in separate glass vials and were dehydrated
with aid of sodium sulfate and magnesium sulfate
salts. In order to obtain the extracts of the samples,
the maceration method was used. About 5.0 g of
the powdered samples were weighed accurately
and added to separate Erlenmeyer asks containing
50 ml methanol (covered with aluminum sheets)
and placed on a magnetic s tirrer for 24 hours. Then,
the extracts were ltered with Watman lter paper
and the solvent was removed by a rotary evaporator
to obtain dry extracts (Schema 1).
2.3. GC-MS Analysis
In the next s tep, the essential oils were subjected
to the GC-MS analysis to separate the chemical
con s tituents and identify them according to their mass
characteri s tics and retention times. The analysis of the
essential oils was carried out using an Agilent sy s tem
equipped with an HP-5S column (30 m × 250 µm,
lm thickness 0.25 µm) interfaced with a quadrupole
mass detector (MS 5977A). Oven temperature 50-
250°C (3°C per minute), injector temperature 250°C,
injection volume: 0.1 μL, split injection with a split
ratio of 1:50 helium as the carrier gas with ow rate
1 mL min-1, ion source: 70 eV, ionization current:
150 μA, and scan range: 35-465. The method was
validated as per the ICH guidelines for linearity,
precision, accuracy, robu s tness, LOD, and LOQ
according to our previous s tudy [16]. Identication
of the chemical con s tituents of the essential oil was
carried out using AMDIS software (www.amdis.net)
and identied by its retention indices with reference
to the n-alkanes series (C6-C20), comparison of their
retention time, mass spectra, and computer matching
with the Wiley 7 nL and NIS t library database.
2.4. Determination of total phenolic content
Total phenol content was determined by the Folin-
Ciocalteu reagent. A dilute solution of the extracts
(0.05:1 g mL-1) or gallic acid ( s tandard phenolic
compound) was mixed with the Folin-Ciocalteu
reagent (2.5 ml, 1:10 diluted with di s tilled water)
and aqueous Na2CO3 (2 ml, 5%). The mixture was
allowed to s tand for 30 min and the phenolic contents
were determined by colorimetry at 765 nm. The total
phenolic content was determined as mg of gallic
Schema 1. Procedure for determination of avonoid, phenolic contents in Ducrosia anethifolia
Anal. Methods Environ. Chem. J. 5 (4) (2022) 66-76
69
acid equivalent using an equation obtained from the
s tandard gallic acid calibration curve [17].
2.5. Determination of total avonoid content
The avonoid contents of the extracts were measured
by aluminum chloride coloration using quercetin as
s tandard [18]. To extract avonoids, 0.1 g of each extract
was solved in 10 mL ethanol 80%. Then, 100 μL of the
solution was added to a te s t tube, and 100 μL of 10%
AlCl3, 100 μL of 1 M sodium acetate, 1.5 mL of ethanol
96%, and 3.2 mL of di s tilled water were added and
vortexed for 1 minute. The control treatment included
3.4 mL of di s tilled water, 100 μL of 1 M sodium
acetate, and 1.5 mL ethanol 96%. After 30 minutes,
adsorption was read at 415 nm.
2.6. DPPH free radical scavenging activity
This spectrophotometric method was used to evaluate
the antioxidant activity of the extracts. DPPH is
a reagent that measures free radical scavenging
activity [19]. Zero, 0.01, 0.02, 0.03, 0.04, 0.05 mL of
concentration 2000 mg L-1 of the extracts and positive
control (ascorbic acid) were added to 1.0 mL of 0.1
mM solution of DPPH (Sigma, S t Louis) in methanol.
The reaction mixture was shaken and then incubated
for 30 min at room temperature. The remaining
amount of DPPH was determined at 517 nm again s t
a blank using a spectrophotometer (Milton Roy
Company Spectronic 2OD). All te s ts were carried out
ve times.
2.7. Ferric-reducing antioxidant power (FRAP)
assay
The antioxidant capacity of the plant extracts was
done by Iron reduction (FRAP assay) according to
Sadeghi et al [17]. For this purpose, 300 mM acetate
buer (pH 3.610 ) mM TPTZ solution in 40 mM HCl,
and 20 mM FeCl3-6H2O solution were mixed for the
preparation of s tock. FRAP reagent was prepared
right away before analysis by mixing 25 mL acetate
buer, 2.5 mL TPTZ solution, and 2.5 mL FeCl3-
6H2O solution. Plant extracts (1000 μg mL-1) were
prepared. 200 μg mL-1 of the extracts was mixed with
1.8 mL of the FRAP reagent and was incubated at
37 ºC for 30 min in the dark condition before being
used. Then, readings of the colored products (ferrous
tripyridyltriazine complex) were determined at 595
nm again s t a di s tilled water blank. FeSO4-7H2O (100-
1000 μM) was used for calibration. Ascorbic acid was
used as a positive control. Results are expressed as
mM Fe2+ per mg sample [17].
3. Results
3.1. Chemical composition of the essential oils
Table 1 shows the major components of the essential
oil in the dierent treatments and control along
with the percentage of each compound. The results
revealed that ethanol spraying had a signicant eect
on the amount and yield of the essential oils. The yield
of the essential oil was enhanced by increasing the
amount of alcohol from 10% to 40% treatment with a
decline in 80% alcohol-treated samples. The chemical
composition of the essential oil of the control and that
of the 10% treatment were to some extent similar.
Intere s tingly, while in the two treatments 20% and
40%, the chemical composition is rather the same, their
chemical composition is dierent from the blank and
10% treatment. Although the chemical composition
of the 80% treatment was similar to 20% and 40%,
there were some dierences. For in s tance, compound
2-isopropyl-5-methyl-3-cyclohexene-1-one was only
identied in the 80% treatment. Decanal, cis-verbenyl
acetate, and dodecanal were major components in all
samples with variations in dierent treatments.
Table 2 shows that monoterpenes and other compounds
including alkanes are dominated in the essential oil
samples. The number of oxygenated monoterpenes
is increased in 20%, 40%, and 80% treatments while
hydrocarbon monoterpenes are decreased in these
samples compared to the control. Hydrocarbon
monoterpenes are absent in the 20% sample.
The number of oxygenated sesquiterpenes is almo s t
the same in all treatments with a small increase of
20% and 40%. The highe s t number of hydrocarbon
sesquiterpenes were observed for 20% sample.
3.2. Determination of total avonoid content
The highe s t and the lowe s t amount of total phenol
per 1g of the plant powder were observed for 40%
Determination and evaluation of the aromatic essential oil in plant Aliyeh Sarabandi et al
70
Table 1. The major compounds identied in the essential oil of the dierent treatments.
aRt bRI Compound Control 10% 20% 40% 80%
4.936 939 α-Pinene 5.272 4.678 - - 0.564
5.52 975 Sabinene 0.936 0.876 - - -
5.589 979 β-Pinene 0.251 0.223 - - -
5.737 990 β-Myrcene 1.428 1.341 - - -
6.287 1024 ρ-Cymene 4.217 3.593 - - 0.330
6.355 1029 d-Limonene 5.178 4.575 - - 0.348
7.271 1088 Terpinolene 0.343 0.329 - 0.523 -
7.46 1100 Nonanal 0.849 1.018 0.425 - 1.057
8.135 1137 cis-Verbenol 0.809 0.682 0.779 - 0.927
8.204 1140 Citronellal 0.801 0.902 - - -
8.364 1150 trans-Verbenol 0.247 0.210 0.962 0.607 0.253
8.432 1169 1-Nonanol 0.732 0.783 - - 0.710
8.713 1179 ρ-Cymen-8-ol 0.458 0.419 - - 0.620
8.776 1185 Cryptone 0.606 0.696 - - -
8.965 1200 Decanal 20.044 20.681 9.098 10.596 13.571
9.274 1250 3,7-Dimethyl-2-octen-
1-ol
1.709 1.479 - - 0.353
9.754 1274 cIMC Hexane - - - - 10.191
9.817 1282 cis-Verbenyl acetate 20.598 18.151 36.274 42.415 21.766
9.88 1286 5-Undecanol 1.816 1.560 1.038 0.778 1.311
10.155 1290 Lavandulyl acetate 0.915 0.828 1.562 1.315 0.623
10.367 1298 trans-Pinocarvyl
acetate
- - 0.420 1.093 -
10.395 1306 Undecanal 1.008 1.183 - - 1.131
10.498 1352 Citronellyl acetate 0.659 0.634 2.058 2.255 1.019
11.408 1381 Geranyl acetate 1.387 1.467 0.958 1.243 0.331
11.677 1408 Z-Caryophyllene 0.451 0.575 0.490 - 0.243
11.757 1410 Dodecanal 10.768 11.953 6.777 6.431 11.053
12.072 1420 β-Caryophyllene 1.666 1.731 1.359 1.308 0.351
12.501 1436 γ-Elemene 3.458 3.246 22.994 1.055 2.038
13.25 1510 dCPCP - - - 18.913 16.705
14.029 1578 Spathulenol 1.898 2.096 2.287 1.577 1.417
14.12 1583 Caryophyllene oxide 1.568 1.709 1.304 0.796 1.161
14.252 1612 Tetradecanal 0.342 0.452 0.467 0.425 0.814
14.515 1620 Unknown 2.536 2.689 1.287 1.165 0.604
14.652 1632 γ-Eudesmol 0.264 0.266 2.008 3.832 1.287
14.738 1677 Z-Nerolidyl acetate 0.778 0.833 0.724 0.462 0.225
14.887 1680 β-Eudesmol 0.506 0.576 1.168 - 1.723
14.939 1685 n-Tetradecanol 1.092 1.334 0.990 0.665 1.582
Total 73.176 90.761 95.429 97.454 94.287
aRetention time; bKovats Index
cMC Hexane: 2-Isopropyl-5-methyl-3-cyclohexen-1-one
dCPCP: 2-Cyclopentylidene cyclopentanone
Anal. Methods Environ. Chem. J. 5 (4) (2022) 66-76
71
and 80% ethanol samples, respectively (Fig. 1). The
highe s t and the lowe s t amount of total phenol per 1.0
g of the plant extract were observed for 40% and 10%
ethanol samples, respectively (Fig. 2).
3.3. Determination of total avonoid content
As shown in Figure 3, the 40% ethanol treatment
showed the highe s t amount of avonoid compared
to the re s t of the treatments.
Table 2. The amount of dierent volatile compounds in dierent treatments.
Compounds Control 10% 20% 40% 80%
Oxygenated Monoterpenes 28.189 25.468 43.013 59.524 36.083
Hydrocarbon Monoterpenes 17.625 16.453 - 0.523 1.272
Oxygenated Sesquiterpenes 5.014 5.480 7.491 6.667 5.813
Hydrocarbon Sesquiterpenes 5.575 5.552 24.843 2.363 4.670
Other Compounds 25.883 38.964 18.795 37.808 47.934
Fig. 1. The total phenol content in the plant powder of dierent treatments.
Fig. 2. The total phenol content of the extracts from dierent treatments.
Determination and evaluation of the aromatic essential oil in plant Aliyeh Sarabandi et al
72
3.4. DPPH free radical scavenging activity
The results of DPPH radical-scavenging activity
assay are shown in Figure 4. Considering the large
variation of IC50, the lowe s t antioxidant activity
was observed for 80% treatment compared to the
control. Treatments with 10%, 20%, and 40%
showed almo s t the same antioxidant activity, all
higher than that of the control.
3.5. FRAP assay
All analyzed extracts demon s trated signicant
antioxidant capacities with FRAP te s t. The 40%
treatment showed 53.00 mMFe2+/mg sample
with the highe s t antioxidant activity compared to
reducing power of ascorbic acid (69.00 mMFe2+/
mg sample) (Fig. 5).
Totally, the results from the determination of phenolic
and avonoid contents of the samples as well as
DPPH and FRAP assays revealed that the amount of
these compounds and the antioxidant capacity of the
plant samples were inuenced by the use of ethanol
spraying which was shown in Table 3. By increasing
the concentration of alcohol from 10% to 40%, the
phenol and avonoid contents and consequently the
antioxidant activity reached their maximum rate.
Fig. 3. The total avonoid content of the extracts from dierent treatments.
Fig. 4. DPPH free radical scavenging activities observed
for dierent treatments compared to ascorbic acid as positive control
Anal. Methods Environ. Chem. J. 5 (4) (2022) 66-76
73
4. Discussion
Foliar feeding is a technique of feeding plants by
applying liquid fertilizer directly to the leaves. Due
to the increased rate of absorption through the aerial
parts of plants, it is an excellent method to deliver
food and elements required for plants much fa s ter
[19]. S tudies show that using alcohols with dierent
concentrations would exert dierent eects on
dierent plant species. The mo s t important role
for methanol operating in C3 plants is to prevent
light respiration, probably due to increased CO2
concentration in leaves. If the concentration of CO2
increases in leaves, ribulose 1,5-bisphosphate will
react with CO2 in s tead of O2, and the carboxylation
function will occur. Therefore, the alcohol-induced
biomass increase of the C3 plants might cause the
plant to use methanol as a direct source of carbon
for serine biosynthesis and reduce carbon wa s te
through light respiration [15]. There are several
reports inve s tigating the eect of alcohol on the
function of dierent plants. For in s tance, Zbiec and
Podsiad (2003) inve s tigated the eect of alcohol
spraying and reported the increasing quantitative
and qualitative yield of this technique on plants such
as geranium, wheat, turnip, and sugar beet [21].
Iqbal Makhdum et al. s tudied the eect of methanol
spraying on cotton plants and observed that 30%
methanol treatment has been able to increase plant
function compared with the control treatment [22].
In another research, performed by Safarzadeh
Vishkaei who s tudied the eect of methanol on
peanuts, 30% methanol treatment could increase
the height of plant and grain function. Methanol
and ethanol (30%) spraying could increase plant
growth and the essential oil amount of peppermint
[23]. The application of alcohol foliar feeding
Fig. 5. The antioxidant capacity of dierent treatments compared
to ascorbic acid as a positive control
Table 3. Comparing the phenolic and avonoid contents as well as antioxidant activities of the samples.
Plant samples IC50
Fe2+ mM/
mg sample
mg total phenol/
1 gr powder
mg total phenol/
1 gr extract
mg quercetin/
1 gr extract
Control 2.87 0.69 5.33 20.44 21.7
10% 66.9 0.43 5.97 19.04 27.94
20% 30.22 0.45 6.71 20.58 30.62
40% 28.55 0.47 9.88 22.12 33.3
80% 91.73 0.44 4.43 18.46 26.54
Determination and evaluation of the aromatic essential oil in plant Aliyeh Sarabandi et al
74
would induce increasing the production of cytokinin
and plant growth [24]. In addition, foliar feeding
of alcohols in plants might induce increasing the
plant metabolites including essential oils. S tudies
show that plants exposed to environmental s tresses
might increase the production of their specialized
metabolites to confront the s timulant leading to
more metabolite synthesis.
The current s tudy showed that alcohol foliar feeding
had eects on the amount of essential oil of the
medicinal plant D. anethifolia and its composition.
Our results showed that monoterpenes and alkanes
such as n-decanal were the major components in
the essential oil of D. anethifolia. In mo s t of the
s tudies inve s tigating the essential oil composition
of this medicinal plant, n-decanal was reported as
the main con s tituent [25,26]. For example, Salari et
al. have reported n-decanal (22.29%) as the mo s t
abundant con s tituent of D. anethifolia essential oil
(Kerman), followed by alkanes decanol (22.18%)
and dodecanol (11/79%) [27]. There are also some
s tudies in which decanal has not been reported
as the major compound [3,28,29]. The results of
Arbabi et al. showed that cis-chrysanthenyl acetate
with an average amount of 44.77% was the main
compound of the essential oil of D. anethifolia
from Si s tan & Baluchi s tan [2].
Besides, our results revealed that this technique
had benecial eects on the amount of phenolic
and avonoid contents and consequently on the
antioxidant activity of the plant. In total, the be s t
eciency was observed for 40% ethanol treatment.
This can be due to the nutritional role of ethanol
as a carbon source, s timulus, and active sub s tance
in metabolic reactions. In addition, ethanol plays
an essential role in the biosynthetic pathway for
the production of terpenoids. The opposite eect
observed in the higher percentages of alcohol might
be due to the plant poisoning leading to the decreased
level of specialized metabolites and antioxidant
capacity. The results of this experiment showed
that the use of the hydroalcohol spraying could
increase essential oil production, so it is sugge s ted
to use of hydroalcoholic foliar feeding to increase
D. anethifolia metabolites in future research.
5. Conclusion
Taken together, the results of this experiment
showed that ethanol spraying could increase the
production of essential oil as well as phenolic and
avonoid compounds in D. anethifolia. Since the
plant is an important medicinal plant, this might
increase its ecacy. Thus, hydroalcoholic foliar
feeding can be a valuable s trategy to increase the
specialized metabolites of D. anethifolia in future
research.
6. Acknowledgements
The authors would like to thank the University of
Si s tan and Baluche s tan for their help and support
(Grant No. 2376081).
7. References
[1] J. Mottaghipisheh, A. Boveiri Dehsheikh,
M. Mahmoodi Soure s tani, T. Kiss, J.
Hohmann, D. Csupor, Ducrosia spp., rare
plants with promising phytochemical and
pharmacological characteri s tics: An updated
review, Pharmaceuticals (Basel), 13 (2020)
175. https://doi.org/10.3390/ph13080175
[2] M. Arbabi, H. Naghdi Badi, M. Labba,
A. Mehrafarin, E. Saboki, Inve s tigating
the essential oil composition of Ducrosia
anethifolia (DC.) Boiss. in dierent altitudes
of Si s tan and Baluche s tan province, Iran, J.
Med. Plants, 19 (2020) 343-355. https://doi.
org/10.29252/jmp.19.74.343
[3] J. Mottaghipisheh, M.T. Maghsoudlou,
J. Valizadeh, R. Arjomandi, Antioxidant
activity and chemical composition of the
essential oil of Ducrosia anethifolia (DC.)
Boiss. from Neyriz, J. Med. Plant. By-prod.,
2 (2014) 215-218. https://doi.org/10.22092/
JMPB.2014.108737.
[4] E.R. Elsharkawy, E.M. Abdallah, M.H.
Shiboob, S.M. Alghanem, Phytochemical,
antioxidant and antibacterial potential of
Ducrosia anethifolia in northern border
region of Saudi Arabia, J. Pharm. Res. Int., 31
(2019) 1-8. https://doi.org/10.9734/jpri/2019/
v31i630361
Anal. Methods Environ. Chem. J. 5 (4) (2022) 66-76
75
[5] N.M. Shalaby, H.I. Abd-Alla, H.F. Aly,
M.A. Albalawy, K.H. Shaker, J. Bouajila,
Preliminary in vitro and in vivo evaluation of
antidiabetic activity of Ducrosia anethifolia
Boiss. and its linear furanocoumarins,
Biomed. Res. Int., 2014 (2014) 480545.
https://doi.org/10.1155/2014/480545
[6] M. Zamyad, M. Abasnejad, S. Esmaeili-
Mahani, A. Mo s tafavi, Alpha-Pinene as the
main component of Ducrosia anethifolia
(Boiss) essential oil is responsible for its
eect on locomotor activity in rats, Avicenna
J. Neuro. Psycho. Physiol., 3 (2016) 29-34.
https://doi.org/10.17795/ajnpp-38787.
[7] M. Akaberi, Z. Tayarani-Najaran, I.
Mehregan, J. Asili, A. Sahebkar, M.
Hassanzadeh-Khayyat, S.A. Emami,
Review of the essential oil composition of
Iranian endemic and native taxa of Apiaceae
(Umbelliferae), Curr. Org. Chem., 24 (2020)
909-1009. https://doi.org/10.2174/13852728
24999200513103632.
[8] M. Abbasnejad, A. Mo s tafavi, R. Kooshki,
P. Hamzenejad, S. Esmaeili-Mahani, Eect
of Ducrosia anethifolia (Dc.) Bioss essential
oil on spatial learning and memory in rats,
J. Gorgan Univ. Med. Sci., 18 (2017) 9-15.
http://goums.ac.ir/journal/browse.php?a_
id=2949&sid=1&slc_lang=en
[9] L. Obeidi, A.A. Mehrabi, M. Omidi, A.
Oladzad, Meiotic behavior and pollen
viability of Ducrosia anethifolia (DC), Iran.
J. Rangelands For. Plant Breed. Genet. Res.,
20 (2012) 124-133. https://doi.org 10.22092/
IJRFPBGR.2012.6631
[10] M. Zamyad, M. Abbasnejad, S. Esmaeili-
Mahani, A. Mo s tafavi, V. Sheibani, The
anticonvulsant eects of Ducrosia anethifolia
(Boiss) essential oil are produced by its
main component alpha-pinene in rats, Arq.
Neuropsiquiatr., 77 (2019) 106-114. https://
doi.org/10.1590/0004-282X20180147.
[11] M. Karami, F. Ghassemi, Eects of aqueous-
alcoholic extract of Ducrosia anethifolia
(DC.) Boiss. leaves on fetal heart tissue
in diabetic rats, Iran. J. Med. Aromatic
Plants Res., 37 (2021) 52-64. https://doi.
org/10.22092/IJMAPR.2021.342950.2791
[12] N. Rahimi, E. Samani Jahromi, S. Zolghadri
Jahromi, The eect of the hydro-alcoholic
extract of (Ducrosia anethifolia) on
te s to s terone hormone and the hi s tological
changes of the te s ticle in male adult rats,
Armaghane-danesh, Yasuj Univ. Med. Sci. J.,
21 (2016) 682-693. https://armaghanj.yums.
ac.ir/article-1-1389-en.pdf
[13] L. Shooshtari, M. Omidi, E. Majidi, M. R.
Naghavi, M. Ghorbanpour, A. Etminan,
Assessment of somaclonal variation of
regenerated Ducrosia anethifolia plants using
AFLP markers, J. Hortic. For. Biotechnol.,
17 (2013) 99-106. https://journal-hfb.usab-
tm.ro/romana/2013/Li s ta%20Lucrari%20
PDF/Lucrari%20Vol%2017(4)%20
PDF/22Lia%20Shooshtari.pdf
[14] C. Wang, L. Guo, Y. Li, Z. Wang, Sy s tematic
comparison of C3 and C4 plants based on
metabolic network analysis, BMC Sy s t. Biol.,
6 (2012) S9. https://doi.org/10.1186/1752-
0509-6-S2-S9
[15] S. Sajedi Moghadam, A. Mehrafarin, H.
Naghdi Badi, A.R. Pazoki, N. Qavami,
Evaluation of phytochemical yield of thyme
(Thymus vulgaris L.) under foliar application
of hydroalcohols, J. Med. Plants, 4 (2012)
130-140. https://doi.org/20.1001.1.2717204.
2012.11.44.12.4
[16] J. Asili, Z. Tayarani-Najaran, S.A. Emami, M.
Iranshahi, A. Sahebkar, S. Eghbali, Chemical
composition, cytotoxic and antibacterial
activity of essential oil from aerial parts of
Salvia tebesana Bunge, J. Essen. Oil-Bear.
Plants, 24 (2021) 31-39. https://doi.org/10.1
080/0972060X.2021.1886996
[17] Z. Sadeghi, J. Valizadeh, O. Azyzian
Shermeh, M. Akaberi, Antioxidant activity
and total phenolic content of Boerhavia
elegans (choisy) grown in Baluche s tan,
Iran, Avicenna J. Phytomed., 5 (2015) 1-9.
https://www.ncbi.nlm.nih.gov/pmc/articles/
Determination and evaluation of the aromatic essential oil in plant Aliyeh Sarabandi et al
76
PMC4352527/
[18] E.V. Beketov, V. P. Pakhomov, O.V.
Ne s terova, Improved method of avonoid
extraction from bird cherry fruits, Pharm.
Chem. J., 39 (2005) 316-318. https://doi.
org/10.1007/s11094-005-0143-7
[19] M. Jahani, M. Akaberi, M. Hasanzadeh
Khayyat, S.A. Emami, Chemical composition
and antioxidant activity of essential oils from
Cupressus sempervirens. var. sempervirens,
C. sempervirens cv. Cereiformis and C.
sempervirens var. horizentalis, J. Essent. Oil-
Bear. Plants, 22 (2019) 917-931. https://doi.
org/10.1080/0972060X.2019.1646672
[20] H. Nourafcan, Z. Kalantari, The eect of
methanol and ethanol foliar application
on peppermint morpho-physiological
charactri s tics, Agroecol. J., 12 (2017) 1-9.
http://www.Agroecology Journal.com
[21] I. Zbiec, C.O. Podsiad, Response of some
cultivated plants to methanol as compared
to supplemental irrigation, Electron. J. Pol.
Agric. Univ., 6 (2003) 1-7. http://www.ejpau.
media.pl
[22] M.I. Makhdum, M.N.A. Malik, S. Din, F.
Ahmad, F.I. Chaudhry, Physiological response
of cotton to methanol foliar application, J.
Res. Sci., 13 (2002) 37-43. https://www.bzu.
edu.pk/jrscience/vol13no1/5.pdf
[23] M.N. Safarazade Vishgahi, G. Nourmohamadi,
H. Magidi, H. Eect of methanol on peanut
function and yield components, Iran. J. Agric.
Sci. 103 (2007) 88. https://jijas.ut.ac.ir
[24] H. Hadadi, P. Moradi, F. Matlabi, The Eect
of methanol and manganese salt spraying on
the amount and components of essential oil
of melissa (Melissa ocinalis L.), J. Med.
Plants, 2 (2016) 80-88. https://doi.org/20.10
01.1.2717204.2016.15.58.13.1
[25] V. Hajhashemi, M. Rabbani, A. Ghanadi,
E. Davari, Evaluation of antianxiety and
sedative eects of essential oil of Ducrosia
anethifolia in mice, Clinics, 21 (1985)
5103-5121. https://doi.org/10.1590/S1807-
59322010001000020
[26] F. Sedkon, I. Javidtash, Essential oil
composition of Ducrosia anethifolia (DC.)
Boiss. from Iran, J. Essential Oil Res., 14
(2002) 278-279. https://doi.org/20.1001.1.27
17204.2020.19.74.24.2
[27] S. Salari, M. Shamsaddini, The production of
nanoliposomal sy s tem containing Ducrosia
anethifolia (DC.) Boiss essential oil made
by sonication and ltration methods, J. Med.
Plants, 19 (2020) 229-238. https://doi.org/20.
1001.1.2717204.2020.19.74.13.1
[28] Z. Karimi, A. Ghani, S. Mohtashami,
Comparative s tudy of essential oil content
and composition of Ducrosia anethifolia at
two phenologocal s tage, Res. J. Pharm., 44
(2017) 29-30. http://rjpharmacognosy.ir
[29] M. Keshavarzi, A. Sharifan, S.A. Yasini
Ardakani, Eect of the essential oil of
Ducrosia anethifolia (DC.) Boiss. and
Teucrium polium L. on physicochemical,
sensory, and microbial characteri s tics of
probiotic yogurt during s torage time, Iran.
J. Food Sci. Technol., 18 (2021) 265-276.
https://doi.org/10.52547/fsct.18.117.265
Anal. Methods Environ. Chem. J. 5 (4) (2022) 66-76
