Essential oil content, composition and antioxidant activity of the root of Smyrnium

cordifolium Boiss. from Southwest Iran

Sadegh Dodmana, Karamatollah Saeidia*, Zahra Lorigooinib, Mahmoud Kianic

a
Department of Horticulture, Faculty of Agriculture, Shahrekord University, Shahrekord, Iran; bMedical
Plants Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences,
Shahrekord, Iran; cDepartment of Horticulture, Faculty of Agriculture, Amol University, Amol, Iran.

Corresponding Author: saeidi@[Link]

Dr. Karamatollah Saeidi (Corresponding author)
Associate Professor of Medicinal and Aromatic Plants, Horticultural Science Department, Faculty of
Agriculture, Shahrekord University, Shahrekord, Iran.
SHORT COMMUNICATION
Essential Oil Content, Composition and Antioxidant Activity of the Root of Smyrnium
cordifolium Boiss in Southwest of Iran.
Sadegh Dodmana, Karamatollah Saeidia*, Zahra Lorigooinib, Mahmoud Kianic
a
Department of Horticulture, Faculty of Agriculture, Shahrekord University, Shahrekord, Iran; bMedical
Plants Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences,
Shahrekord, Iran; cDepartment of Horticulture, Faculty of Agriculture, Amol University, Amol, Iran

Abstract
In this research, root samples of ten ecotypes of Smyrnium cordifolium Boiss. were
collected from their natural habitats throughout southwest of Iran. Essential oil (EO)
samples were obtained by using a clevenger-type apparatus, and were then chemically
characterized by gas chromatography (GC) and gas chromatography with mass
spectrometry (GC–MS) methods. EO yields ranged from 0.13-0.74% (w/w). EO
samples were predominantly constituted by germacron, curzerene and curzerenone.
Revealed by the hierarchical cluster analysis, the examined ecotypes were nested in
different groups in relation to intrinsic factors and genetic makeup rather than extrinsic
elements. Antioxidant activity of the samples was evaluated using DPPH radical
scavenging method (0.33-8.59 ug/ml). The current research provided new insights into
the chemical variation of Smyrnium together with a possible application of its essential
oil as a natural preservative agent in food and pharmaceutical industries.
Keywords: Smyrnium cordifolium; Essential oil; Aantioxidant activity; Iran
1. Introduction
Smyrnium is a genus of flowering plants in the family Apiaceae, indigenous to central Asia, Iran,
Turkey, and western Afghanistan. The genus is represented in the flora of Iran by only one species,
Smyrnium cordifolium. The species is well-adapted to the uplands, foothills and associated plains
of the Zagros range in the northwest (Esmaili & Amiri 2006)) and southwest regions of Iran
(Mehrabi & Mehrabi 2011).
Nearly all parts of the plant including stem, leaves, fruits and roots contain essential oil,
predominantly constituted by the compounds belonging to different classes of terpenes e.g.
curzerene, curzerenone and germacrene-D. In the Iranian traditional medicine, Smyrnium
cordifolium has found application as an e.g. antifungal, antibacterial, diuretic, restorative and
counteracting renal calculus agent(Ulubelen et al. 1985; Amiri et al. 2006; Ahvazi et al. 2011).
Additionally, the fleshy stems of the plant are used as vegetables (Amiri et al. 2006).
Oxidation is known to have negative effects, especially in industrial context. Oxidation reactions
occur in foods and cosmetics, often because of the prolonged exposure to oxygen. Food
preservation represents an issue of concern and requires assurance of protection from microbial
spoilage and prolongation of shelf life (Mimica-Dukić et al. 2003). The damaging effects caused
by oxidation have commonly been reduced by the usage of synthetic antioxidant molecules that
scavenge free radicals and reduce/prevent their damages. However, health critics and researchers
are questioning the safety of such compounds, as the usage of synthetic chemicals has increasingly
become viewed as a threat to human health. Plant-derived compounds have gained considerable
importance due to their increasing applications as additives in food and cosmetics technologies for
the protection of easy oxidizing components (Gruľová et al. 2012). Therefore, identification of
natural antioxidants as preservative agents plays a pivotal role in the food, cosmetic and
pharmaceutics industries (Saleh et al. 2010).
To date, very few attempts have been made to investigate the essential oil of the Iranian Smyrnium,
and we know little about its composition and antioxidant properties. Moreover, all of these efforts
are confined to a limited geographical area. To address the information gaps mentioned above, in
the current study, the variation in the essential oil yield, chemical constituents and antioxidant
activity among ten Iranian populations of S. cordifolium was investigated.
2. Results and discussion
2.1. Essential oil analysis
The yield in EO of ten smyrnium ecotypes was evaluated and results are summarized in (Table
1). According to our findings, The yield in EOs ranged from 0.13% for Farsan ecotype (SC10)
(SC8) to 0.74% for for Soodejan ecotype (Table 1). The essential oil yield of root samples from
the highest to the lowest was as follows: Farsan ecotypes (0.74%), Shah Qasem dam (0.53%),
Darshahi (0.49%), Gol zardi (0.47%), Gorgo (0.34%), Mukhtar (0.32%), Deh Bar Aftab (0.20%),
Lordegan (0.24%), Mahmoud Abad (0.21%), %) and Soodejan (0.13%). In previous studies, the
yield of essential oil of smyrnium was reported to be between 0.7% and 7.7% (Amiri et al. 2007).
The composition of SEOs was also analyzed and qualitative and quantitative differences were
observed. According to the results, the minor and major EO components were variable in
occurrence and concentration among the tested samples. In total, 18 compounds were identified in
all of the specimens, accounting for over 98.60-99.48% of the total oil. The highest number of
compounds was recorded in SC1 with 18 compounds, followed by SC2 (17 compounds), SC6 (16
compounds), SC10 (15 compounds), SC3 and SC5 (11 compounds), SC7, SC8 and SC9 (12
compounds), and SC4 (11 compounds). In general, the majority of the compounds identified
belonged to different classes of terpenes including monoterpenes (0-1.64%), oxygenated
monoterpenes (0-10.47%), sesquiterpenes (40.97-60.95%) and oxygenated sesquiterpenes (30.86-
57.74%), of which sesquiterpenes boast the highest total contribution percentage of the total oils.
Generally, the SEOs were predominantly constituted by germacrene, curzerene and curzerenone,
respectively (Table 2). The highest and the lowest contents of germacrene were recorded in SC9
(38.40%) and SC4 (58.17%), respectively. SC3 and SC9 contained the highest (47.04%) and the
lowest (23.96%) concentrations of curzerene, while SC10 (3.19%) and SC7 (7.10%) yielded the
highest and the lowest contents of curzerenone. In addition to these three major compounds, an
unknown compound (8.2%) in SC1, thymol (3.81%, 6.84%) and carvacrol (3.35%, 3.25%) in SC3
and SC6, respectively, and Caryophyllene oxide (2.12%) in SC9 were the constituents above 2%
of concentration.
Amiri et al. (2007) characterized the essential oil composition of Smyrnium cordifolium from
northwestern Iran, and results were as follows: The major constituents of the leaf and root EOs
were two oxygenated sesquiterpenes (curzerenone and curzerene), while a reversed order was
found in the stem and fruit oils. Hexadecanoicacid was another major compound in the stem and
leaf oil samples. In another study, Esmaeili and Amiri (2006) reported curzerene and curzerenone,
followed by germacrene-D, isopimarol and phyllocladanol as the major components of the
essential oil of Smyrnium cordifolium (Esmaili & Amiri 2006). The oils from the hypogeous
(below ground) and epigeous (above ground) parts of Smyrnium perfoliatum have also been
reported. The major components were found to be α-pinene (39.3%), aromadendrene (13.2%) and
α-terpinyl valerate (10.0%) in the hypogeous part oil and aromadendrene (49.9%), neryl
isovalerate (10.2 %), δ-muurolene (4.9%) and α-santalene (4.9%) in the epigeous part oil. In the
fruit oil of the plant, germacrene D (45.2%) was the main constituent. The root, herbage and flower
oils of S. olusatrum have been characterized by a high content of oxygenated sesquiterpenoids,
most of which were furano sesquiterpenoids, while the oil of green and ripe fruits of the plant were
dominated by monoterpene hydrocarbons with β-phellandrene and α-pinene as major constituents
(Tirillini et al. 1996). Another study identified several compounds e.g. myrcene, B-flandonor, B-
caryophyllene-furanodine, curzerenone, germacron B and germacron in the root, stem and leaf
EOs of S. olusatrum (Bertoli et al. 2004).In the previous researches carried out on this species, the
sample collection was made from a limited geographical area, while in the present study samples
were collected from a wider area and climatic conditions. It is widely accepted that the production
of EOs in plants is a complex interplay of intrinsic and extrinsic elements. This is
particularly modulated at the DNA level, where specific enzymes in the biosynthetic pathway of
desired products are encoded (Kurosaki. 2012). Therefore, the genetic structure can create
substantial variability in occurrence and concentration of active substances in plants (Kiani et al.
2015). On the other hand, metabolism of SMs is under the influence of extrinsic elements like e.g.
agroclimatic factors as well. Influenced by environmental elements, each class of SMs functions
as a chemical interface between the environment and the plant. These phytochemicals exert their
biological roles, as plastic adaptive drivers in response to the surrounding environment. Such
chemical interactions often include variations in the production of plant metabolites, which could
possibly be one of the main explanations behind plants metabolic diversity (Kiani et al. 2015).

2.2. Cluster analysis

To further explore the chemotaxonomic affinity and relationship among ten Smyrnium ecotypes,
the relative percentage of all analytes were submitted to a hierarchical cluster analysis. Results
indicated that Smyrnium ecotypes diverged across the tree in 3 main groups (Fig. 2). SC4 (Gol
Zardi), SC7 (Mahmood Abad), SC10 (Farsan) and SC3 (Mokhtar) specified with fairly similar
contents of curzerene (~ 31%), germacrene (~ 56%) and curzerenone (~ 5%). The second clade
(Group II) was further diverged to place SC2 (Shah Qassim Dam) and SC8 (Soodejan), SC1
(Gorgo) and SC6 (Deh Bar Aftab) in one group. contents of germacrene )Above 45 lower than
56%) . SC9 (Lordegan) and SC5 (Darshahi) with germacrene Below 45%) Were in the third group.
The least content of germacrene was found in these ecotypes respectively as compared with other
ecotypes. In addition, SC9 was the only ecotype in which the content of curzerene was greater than
that of germacrene. In a general view, the major constituents of the EOs including curzerene,
germacrene and curzerenone played more important role in separation of the ecotypes. Our data
revealed that the examined specimens were scattered across the tree with no evident relationships
to bioclimatic zone or geographical origin. The specimens were nested in different groups in
relation to intrinsic factors and genetic makeup rather than extrinsic elements.

2.2. Antioxidant activity

The free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging assay (Burits & Bucar 2000).
was carried out for determining the antioxidant capacity of SEO samples with reference to the
concentrations providing 50% inhibition (IC50 value). DPPH is a stable organic free radical with
an absorption band at 517 nm. It loses this absorption when accepting an electron or a free radical
species, which results in a visually noticeable discoloration from purple to yellow. It can
accommodate many samples in a short period and is sensitive enough to detect active ingredients
at low concentrations (Do et al. 2014). The antioxidant capacity of a given compound is inversely
related to its IC50, which simply implies that the lower IC50 of a certain chemical, the greater
antioxidant activity of the compound (Do et al. 2014). The addition of SEO to the DPPH solution
induced a rapid decrease in the optical density at 517 nm, and resulted in the significantly better
free radical scavenging activity (lower IC50 value). SC2 was the most potent DPPH scavenger
(0.33±030 μg/ml), followed by SC10 (0.45±0.050 μg/ml), the activity of which was comparable
to that of the superior sample. In this assay, SC7 showed the minimal scavenging activity, as it
removed the DPPH radical at the highest concentration (8.59±0.160 μg/ml). Smyrnium has been
shown to be a potent natural antioxidant source (Khanahmadi et al. 2010; Tabaraki & Ghadiri
2013). Likewise, results of our study revealed high DPPH activity of SEOs. Generally, the
antioxidant capacity of EOs largely depends on the composition of the EO. Therefore, all the
aforementioned factors (e.g. genetic makeup and agroclimatic conditions) influencing the
composition of EOs, would accordingly affect the antioxidant capacity of plants. A large number
of medicinal plants contain chemicals with antioxidant activity, among which phenolics and their
derivatives, e.g. thymol and carvacrol, are receiving particular attention (Dorman et al. 2003). A
highly positive relationship between total phenols and antioxidant activity has been found in many
plant taxa. Although the exact mode of action of phenolics has not been well understood, such
compounds may have effect on biological systems due to their ability to scavenge free
radicals, donate hydrogen atoms or electron, or chelate metal cations. For example, the phenolic
moiety in flavonoids can accept an electron to form relatively stable phenoxyl radicals, thereby
disrupting chain oxidation reactions in cellular components (Shamloo et al. 2017). This might also
be associated with the free radical scavenging activity of a variety of enzymes namely superoxide
dismutase, catalase, peroxidase, etc. as well as the presence of carotenoids, tocopherol and ascorbic
acid. The synergistic effects of other phytochemicals should also be taken into account.
Disclosure statement
No potential conflict of interest was reported by the authors.

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Table 1
Geographical distribution and essential oil content of Smyrnium cordifolium Boiss. populations
throughout southwest Iran.
code origin Latitude Longitude Elevation (m EO content
a.s.l.) )%(
SC1 Gorgo 30° 27' 51° 40' 2269 0.34
SC2 Shah Qasim 30° 34' 51° 33' 2034 0.53
Dam
SC3 Mokhtar 30° 40' 51° 31' 2011 0.32
SC4 Gol zardi 30° 29' 51° 29' 2229 0.47
SC5 Darshahi 30° 51' 51° 20' 1679 0.49
SC6 Deh Bar Aftab 30° 45' 51° 32' 2230 0.30
SC7 Mahmoud 30° 39' 51° 36' 1933 0.21
Abad
SC8 Soodejan 32° 31' 50° 24' 2248 0.13
SC9 Lordegan 31° 29' 51° 00' 1982 0.24
SC10 Farsan 32° 15' 50° 34' 2022 0.74
 Table 2. Essential oil composition of Smyrnium cordifolium Boiss. populations
Ecotypes
Component RI SC1 SC2 SC3 SC4 SC5 SC6 SC7 SC8 SC9 SC10
α-pinene 933 1.26 0.77 0.55 0.29 0.29 0.13 0.38 - 0.12 0.38
β-pinene 978 0.19 0.14 - - - - - - 0.19 0.26
Borneol 1170 0.12 0.23 - - - 0.39 - - - -
Thymol 1303 0.87 0.80 3.81 - 0.13 6.84 - 1.15 - 0.09
Carvacrol 1312 0.70 0.22 3.35 - 0.10 3.25 - 0.74 - 0.01
β-Elemene 1391 0.83 1.19 1 0.89 1.08 0.83 0.89 0.83 1.30 0.90
Curzerene 1500 31.11 35.16 23.96 31.19 34.74 30.34 31.30 38.86 47.04 32.23
Germacrene A 1508 0.24 0.42 0.92 0.23 0.13 0.58 0.71 0.09 0.19 0.13
δ-Cadinene 1519 0.42 0.43 0.58 0.24 0.13 0.56 0.73 - 0.16 0.31
Myristicine 1527 0.84 0.57 0.45 - 0.14 0.62 0.30 0.14 - -
Germacrene B 1559 0.42 0.70 0.42 0.57 0.6 0.46 0.34 0.60 0.92 0.68
Curzerenone 1582 5.84 5.55 3.94 5.88 5.84 5.43 7.10 3.96 6.84 3.19
Caryophyllene oxide 1586 0.85 0.61 1.26 0.64 0.66 0.69 1.05 0.23 2.12 0.64
Atractylone 1661 0.57 0.84 0.85 0.54 0.54 0.75 0.85 0.65 0.87 0.39
Germacrene 1669 45.07 51.06 57.58 58.17 43.30 45.83 54.97 51.50 38.40 56.75
Unknown 1700 8.2 - - - - - - - - -
Unknown 1879 1.10 0.58 0.88 0.73 1.82 1.63 0.74 0.64 1 1.68
Unknown 1992 0.33 0.21 - - - 0.44 - - - 0.18
Total - 99/17 99.48 99.55 99.37 98.60 98.77 99.36 99.37 99.15 98.82
Monoterpenes hydrocarbons - 1.45 0.91 0.55 0.29 0.29 0.13 0.38 0 0.31 1.64
Oxygenatedmonoterpene - 1.69 1.25 7.16 0 0.23 10.47 0 1.89 0 0.1
Sesquiterpenehydrocarbons - 47.82 54.38 60.95 60.1 44.88 48.88 57.94 53.16 40.97 58.77
Oxygenatedsesquiterpens - 38.58 43.00 30.86 38.79 51.32 37.96 41.15 44.35 57.74 36.84
Others - 9.63 0.79 0.88 0.73 1.82 2.07 0.74 0.64 1 1.86
Table 3. Antioxidant activity of essential oils from ten populations of Smyrnium cordifolium Boiss.
Population IC50 (µg /ml)
SC1 3.30±0.080 d
SC2 0.33±0.030 a
SC3 2.24±0.100 b
SC4 2.57±0.270 c
SC5 3.47±0.000 d
SC6 3.30±0.025 d
SC7 8.59±0.160 f
SC8 3.80±0.060 e
SC9 2.41±0.115 bc
SC10 0.45±0.050 a
Different letters indicate significant differences between groups (p < 0.05)
Fig. 1. Collection regions of the studied S. cordifolium Boiss populations.
Fig. 2. Smyrnium l0 ecotypes substratum according to the three major combinations of
essential oils based on the ward method.