Composition and Antioxidant Effect of the Essential Oils of the Flowers and Fruits of Ducrosia assadii Alava., a Unique Endemic Plant from Iran
Essential oils are a kind of secondary metabolites with a variety of valuable biological activities. In the present study, the essential oils of the flowering and fruiting tops of Ducrosia assadii Alava. (Apiaceae), were isolated separately by steam distillation and studied by Gas Chromatography (GC) and Gas Chromatography Mass Spectrometry (GC/MS). Antioxidant effect of the essential oils was studied using diphenylpicrylhydrazil (DPPH) scavenging assay. The results indicated some quantitative but not qualitative differences between the analyzed oils. The major components of the oil were found to be n-decanal 46.68 and 42.21%, (n-decanol ) 13.79, 13.27% and (dodecanal) 10.01 and 8.8%, respectively in flowers and fruits. In both studied oils, oxygenated aliphatic compounds comprise the highest percentage of the oil (76.88, 77.71% in flower and fruit oil, respectively) while the sesquiterpenes were at low percentage in both oils. Both these oils exhibited relative antioxidant effect in comparison to butylated hydroxyanizole (BHA).The composition of the essential oil of D. assadii was completely different from the other studied species of the plant such as D. anethifolia and D. esmaeli which could be accounted for their biologic activities.
to cite this article:
Ali Assadipour, Fariba Sharififar, Maryam Robati, Vida Samzadeh and Khadijeh Esmaeilpour, 2013. Composition and Antioxidant Effect of the Essential Oils of the Flowers and Fruits of Ducrosia assadii Alava., a Unique Endemic Plant from Iran. Journal of Biological Sciences, 13: 288-292.
March 15, 2013; Accepted: March 29, 2013;
Published: July 10, 2013
Ducrosia belongs to the family of Apiaceae (Umbelliferae) and comprises
about only 4 known species which occur largely in Africa and Asia especially
in Iran, Afghanistan, Pakistan, Syria, Lebanon and Iraq (Aynehchi,
1990; Zargari, 1996). This genus comprises about 3
habitat species: Ducrosia anethifolia Boiss., Ducrosia flabellifolia
Boiss. and Ducrosia assadii Alava in Iran (Rechinger,
1987). The plant of D. assadii is described by Mozaffarian for a
vast region in Kerman province between Dehbakri and Jiroft, Lalehzar and Baghin
regions and known in Persian as Moshgake bakraei (Mozaffarian,
1998). Phytochemical studies have demonstrated the presence of some coumarins
such as pangelin in D. anethifolia (Starvi et
al., 2003). The flowers and fruits of D. assadii Alava have
been used as a flavoring agent, antispasmodic and carminative in folk medicine.
Janssen et al. (1984) reported α-pinene
(59.2%), myrcene (11.6%) and limonene (7.5%) as the major compounds of the essential
oil of D. anethifolia (Janssen et al., 1984).
In another report, the major compounds of the oil of D. anethifolia were
reported to be α-pinene, terpineolene and ocimene (Mostafavi
et al., 2008). Haghi et al. (2004)
reported the presence of α-pinene, citronellal, limonene, linalool and
myrcene in this oil (Haghi et al., 2004) while
in the another report, n-decanal constitutes more than 70% of the essential
oil which has exhibited antianxiety effect in mice (HajhashemiI
et al., 2010). There are lesser studies for D. assadii in
the literature. A report shows n-decanal (36.4%) as the main component among
the 29 constituents characterized in the oil of D. assadii (Rustaiyan
et al., 2006), meanwhile Mostafavi et al.
(2008) reported that the essential oil of the aerial parts of the plant
is composed of n-decanal (74%), dodecanal (7.2%) and α-pinene (4.0%) (Mostafavi
et al., 2010).
Several studies suggest that free radicals can damage cellular macromolecules
(Bergendi et al., 1999). Some of plant secondary
metabolites especially essential oils exhibit antioxidant effect (Bensky
et al., 2004; Sharififar et al., 2003,
2007a, b, 2009,
2011; Yassa et al., 2005).
As a part of a project on the volatile composition of Ducrosia species
endemic to Iran, we have studied the oil composition of the aerial parts of
D. assadii in two separate stages of flowering and fruiting times by
GC and GC/MS. We think that the knowledge of the essential oils may be useful
to clarify the systematic status of D. assadii. On the other hand the
antioxidant effect of these essential oils has been studied using DPPH free
radical scavenging assay.
METHODS AND MATERIALS
Plant materials: The air-dried tops of D. assadii were gathered
at flowering and fruiting stages (late June and August, 2011, respectively)
from Dehbakri, Kerman, Iran (at altitude of 1900 m above sea level). Voucher
specimen was deposited at the Herbarium of department of Pharmacognosy of Faculty
of Pharmacy in Kerman University of Medical Sciences, Kerman, Iran (KF1387).
Isolation of essential oil: The gathered parts of the plant were left
to dry at room temperature and 100 g of flowering and fruiting tops of the plant
were coarsely minced and placed in a flask containing 500 mL of water and steam
distilled in a Clevenger apparatus for 4 h. The obtained essential oils were
dried over anhydrous sodium sulfate and stored at 4°C in the darkness. Essential
oil yield was 0.71% and 0.49% based on dried weight of sample for flower and
fruit tops, respectively.
Gas chromatography: The essential oils were analyzed using a Shimadzu
QP 5000 gas chromatograph equipped with a FID detector and HP-5 MS capillary
column (30 m*, 0.25 mm, film thickness 0.25 m). Injector and detector temperatures
were set at 220 and 290°C, respectively. Oven temperature was kept at 50°C
for 3 min, then gradually raised to 160 at 3°C min-1, held for
10 min and finally raised to 240 at 3°C min-1. Helium was the
carrier gas, at a flow rate of 1 mL min-1. Diluted samples (1/100
in acetone, v/v) of 1,0 L were injected manually in the splitless mode. Quantitative
data were obtained electronically from area percent data without the use of
Gas chromatography/mass spectrometry: GC-MS analysis of the essential
oils was performed under the same conditions with GC (column, oven temperature,
flow rate of the carrier gas) using a Shimadzu QP 5000 gas chromatograph equipped
with a Shimadzu QP 5050 mass selective detector in the electron impact mode
(70 ev). Injector and MS transfer line temperatures were set at 220 and 290°C,
Qualitative and quantitative analysis: The components of the oils were
identified based on the comparison of their relative retention times and mass
spectra with those of standards. The individual compounds were identified by
MS and their identity was confirmed by comparing their retention indices relatives
to n-alkanes and by comparing their mass spectra and retention times with those
of authentic samples or with data already available in the NIST and Wiley library
and literature (Adams, 1995). GC and GC/MS analysis results
are given in Table 1.
||GC/MS analysis of the essential oil of Ducrosia assadii
in two stage of flowering (FLD) and fruiting (FRD)
DPPH free radical scavenging assay: The antioxidant activity of FLD
and FRD was determined in terms of hydrogen donating ability using the DPPH
free radical scavenging assay as modified by Sanchez-Moreno
et al. (1998). At first a serial dilution of the essential oils was
prepared . A volume of 0.1 mL of each sample was added to 2.9 mL of DPPH solution
(60 μM). The absorbance was read for 30 min at 5 min intervals using a
spectrophotometer. The remaining concentration of DPPH in the reaction medium
was calculated through the linear regression. The percentage of remaining DPPH
was expressed as following formula:
where, DPPH30 is DPPH concentration after 30 min and DPPH0
is the initial concentration. The percentage of DPPH inhibition of different
concentration of essential oils was determined as follow equation:
where, Ablank is the absorbance of the control reaction (containing
all reagents except the test compound) and Asample is the absorbance
of the test compound. Each experiment was performed in triplicate and data was
reported as Mean±SD.
RESULTS AND DISCUSSION
The components of the oils from the flowering and fruiting tops of D. assadii,
their retention indices and percentage composition are summarized in Table
1, where all the components are listed in order of elution on the DB-5 column.
The major constituents of the essential oil of the flowers of D. assadii
(FLD) were found to be n-decanal (46.68%), n-decanol (13.79%), dodecanal (10.01%)
and alpha-pinene (6.14%). Other representative components of the oil were identified
as beta-myrcene (4.4%), decanoic acid (2.27%), linalool (2.17%), limonene (1.55%)
and alpha terpineolene (1.32%). The total amount of aliphatic compounds was
78.15%, one of monoterpenes was 17.49%, whereas the one of sesquiterpenes was
1.05%. The major constituents of the oil from the fruiting tops of D. assadii
(FRD) were found to be n-decanal (42.21%), n-decanol (13.27%) and dodecanal
(8.8%). Other characteristic components of this oil were decanoic acid (5.53%),
beta- myrcene (2.72%), linalool (2.51%), alpha-pinene (2.30%), beta-caryophyllene
(2.8%) and alpha-terpineolene (1.54%). The amount of aliphatic compounds was
77.77% which 77.25% of them were oxygenated. The amount of monoterpenes was
14.76% and one of sesquiterpenes was about 2.8%. These results show that the
composition of essential oils from FLD and FRD are similar qualitatively but
not quantitatively. n-decanal, n-decanol and dodecanal have been identified
as main constituents in both oils. The components such as n-decanal and dodecanal
are currently obtained from synthetic sources and the high percentage of them
in plant sources would be a good source for them. The composition of essential
oils of D. assadii are completely different from the others species
of Ducrosia. As the report of Janssen et al.
(1984) alpha-pinene, citronellal, limonene, linalool and myrcene (Janssen
et al., 1984; Mostafavi et al., 2008)
were presented as main components and n-decanal, n-dodecanal, n-decanol, trans-2-dodecenal
and cis-chrysanthenyl acetate were lesser while as in D. ismaelis, α-pinene,
n-butylbenzene, gamma-cadinene, 3,5-dimethylstyrene, 5-methylindan have been
identified and reported as major compounds (Al-Meshal et
al., 1985). Our results also show some differences with previous studies
for this plant. Rustaiyan et al. (2006) reported
n-decanal (36.4%) as the main compound in the oil of aerial parts of D. assadii
and Mostafavi et al. (2008, 2010)
indicated the amount of n-decanal about 70 and 36.4% in the oil of aerial parts
of the plant collected from Lalehzar and Dehbakri, respectively. These differences
might be due to the environmental factors such as humidity, temperature and
even altitude of natural habitat where this species grows. In DPPH assay, the
degree of DPPH discoloration is attributed to reducing potency of test compounds.
The results have given in Fig. 1. As shown in this figure,
both FLD and FRD essential oils could inhibit DPPH radical in a concentration-dependent
manner. The highest inhibition was at 67.4±3.6% and 68.5±2.9%
for FLD and FRD, respectively.
||Antioxidant effect of the essential oil of Ducrosia assadii
in two stage of flowering (FLD) and fruiting (FRD), FLD: Flower essential
oil, FRD: Fruit essential oil experiment was carried out in triplicate and
results was reported as Mean±SD
||Time-remained DPPH radical study of the essential oil of Ducrosia
assadii in two stage of flowering (FLD) and fruiting (FRD) in comparison
to BHA, FLD: Flower essential oil, FRD: Fruit essential oil experiment was
carried out in triplicate and results was reported as Mean±SD
These results demonstrated the relative effectiveness of FLD and FRD essential
oils in comparison to BHA, however this activity was weaker than BHA. The highest
DPPH inhibition effect of both essential oils was seen at 320 μL mL-1,
so, this concentration was used for time- activity studies. The time- remaining
DPPH kinetic study shows that the concentration of DPPH decreases with time
in the presence of all concentrations of both essential oils (Fig.
2). DPPH concentration was about 38.1±2.6% and 34.3±2.4% for
FLD and FRD at highest used concentration. No significant difference was observed
in antioxidant effects of tested essential oils. Analysis of FLD and FRD oils
indicated the presence 20.27% and 19.39% monoterpenes in these oils, respectively
(Table 1). Monoterpenes with methylene group exhibit potent
antioxidant effect which attributed to the presence of methylene groups in monoterpene
hydrocarbons (Tepe et al., 2005). These oils
contain a low percentages of monoterpenes and so exhibited a moderate antioxidant
effect. Furthermore, it is expected, in the plants exposed to dry climate, the
compounds with high molecular weight impedes it to low humidity, although more
studies are needed for this hypothesis.
Authors are thankful from Dr. Mitadzadini for their kindly help for identification
of the plant.
Adams, R., 1995. Identification of Essentiol Oils Components by Gas Chromatograph/Mass Spectroscopy. Allured Publcation, Illinois.
Al-Meshal, I., T. Khalifa and M. Hassan, 1985. Physico-chemical characteristics and spectroscopy of Ducrosia ismaelis oil. Spectroscopy Lett., 18: 491-496.
Aynehchi, Y., 1990. Pharmacognosy and Medicinal Plants. Tehran University Publishers, Tehran, Iran.
Bensky, D., S. Clavey and E. Stoger, 2004. Materia Medica. Eastland Press Inc., Hong Kong, pp: 193-194.
Bergendi, L., L. Benes, Z. Durackova and M. Ferencik, 1999. Chemistry, physiology and pathology of free radicals. Life Sci., 65: 1865-1874.
Haghi, G., A. Safaei and J. Safari, 2004. Extraction and determination of the main components of the essential oil of Ducrosia anethifolia by GC and GC/MS. Iran J. Pharm. Res., 2: 90-91.
HajhashemiI, V., M. Rabbani, A. Ghanadi and E. Davari, 2010. Evaluation of antianxiety and sedative effects of essential oil of Ducrosia anethifolia in mice. Clinics, 65: 300-302.
Janssen, A.M., J.J.C. Scheffer, A. Baerheim Svendsen and Y. Aynehchi, 1984. The essential oil of Ducrosia anethifolia (DC.) Boiss Chemical composition and antimicrobial activity. Pharm. Weekbl. Sci., 6: 157-160.
PubMed | Direct Link |
Mostafavi, A., D. Afzali and S. Mirtadzadini, 2008. Chemical composition of the essential oil of Ducrosia anethifolia (DC.) Boiss. from Kerman Province in Iran. J. Essention Oil Res., 20: 509-512.
Direct Link |
Mostafavi, A., T. Shamspur, D. Afzali and S. Mirtadzadini, 2010. Chemical composition of the essential oil of Ducrosia assadii Alava. from Kerman Province in Iran. J. Essential Oil Res., 22: 300-302.
CrossRef | Direct Link |
Mozaffarian, V., 1998. A Dictionary of Iranian Plant Names. Farhang Moasern, Tehran.
Rechinger, K., 1987. Flora Iranica. Akademische Druck and Verlagsanstalt, Iran.
Rustaiyan, A., H. Mazloomifar, S. Masoudi and Z. Aghjani, 2006. Volatile oils of Ducrosia assadii alava. and Prangos acaulis (DC.) Bornm. from Iran. J. Essent. Oil Res., 19: 682-684.
Direct Link |
Sanchez-Moreno, C., J.A. Larrauri and F. Saura-Calixto, 1998. A procedure to measure the antiradical efficiency of polyphenols. J. Sci. Food Agric., 76: 270-276.
CrossRef | Direct Link |
Sharififar, F., A. Derakhshanfar, G. Dehghan-Nudeh, N. Abbasi and R. Abbasi et al., 2011. In vivo antioxidant activity of Zataria multiflora Boiss essential oil. Pak. J. Pharm. Sci., 24: 221-225.
Sharififar, F., G. Dehghn-Nudeh and M. Mirtajaldini, 2009. Major flavonoids with antioxidant activity from Teucrium polium. Food Chem., 112: 885-888.
Sharififar, F., M.H. Moshafi, S.H. Mansouri, M. Khodashenas and M. Khoshnoodi, 2007. In vitro evaluation of antibacterial and antioxidant activities of the essential oil and methanol extract of endemic Zataria multiflora Boiss. Food Control, 18: 800-805.
CrossRef | Direct Link |
Sharififar, F., V. Mozaffarian and S. Moradkhani, 2007. Comparison of antioxidant and free radical scavenging activities of the essential oils from flowers and fruits of Otostegia persica Boiss. Pak. J. Biol. Sci., 10: 3895-3899.
CrossRef | PubMed | Direct Link |
Shrififar, F., N. Yassa and A. Shafiee, 2003. Antioxidant activity of Otostegia persica (Labiatae) and its constituents. Iranian J. Pharm. Res., 2: 235-239.
Direct Link |
Starvi, M., K.T. Mathew, F. Bucar and S. Gibbons, 2003. Pangelin, an antimycobacterial coumarin from Ducrosia anethifolia. Planta Med., 69: 956-959.
Tepe, B., D. Daferera, A. Sokmen, M. Sokmen and M. Polissiou, 2005. Antimicrobial and antioxidant activities of the essential oil and various extracts of Salvia tomentosa Miller (Lamiaceae). Food Chem., 90: 333-340.
CrossRef | Direct Link |
Yassa, N., F. Sharififar and A. Shafiee, 2005. Otostegia persica as a source of natural antioxidants. Pharm. Biol., 43: 33-38.
Zargari, A., 1996. Medicinal Plants. Vol. II, Tehran University Publications, Tehran, Iran.