Chemical Composition of the Essential Oil of Artemisia absinthium Growing Wild in Iran
Studies were conducted to investigate the composition
of essential oil of wormwood (Artemisia absinthium L.) growing
wild in Iran. The wormwood aerial parts were harvested in full blooming
time from an area between Deylaman and Asiabar villages, at Alborz altitudes
in Guilan province in September 2005. Aerial parts were dried at shade
(room temperature) for several days and their essential oil was extracted
by hydrodistillation method in a Clevenger apparatus and analyzed by GC/MS.
Results showed that essential oil yield was 1.3%. Twenty eight components
representing 93.3% of the oil were identified, which were mostly monoterpenes.
β- pinene and β- thujone were the main components, which their
contents were 23.8 and 18.6% respectively. The largest part of the essential
oil was formed by hydrocarbon monoterpenes (47.8%). The results proved
that chemotype of the studied wormwood essential oil was specific and
different from other wormwood essential oil chemotypes, which have been
reported so far.
Artemisia absinthium L. is an aromatic plant of the
family Asteraceae, subfamily Asteroideae, tribe Anthemideae and is known
by the common names wormwood (UK), absinthe (France), wermut (Germany)
and afsantine (Iran) (Zargari, 1989; Wright, 2002). The plant grows in
North and East of Iran (Zargari, 1989) and also is very common in Turkey
(Aslan et al., 2005; Kordali et al., 2006). The herb is
native to warm Mediterranean countries, usually found growing in dry waste
places such as roadsides, preferring a nitrogen-rich stony and hence loose
soil. Wormwood has been naturalized in northeastern North America, North
and West Asia and Africa. The plant`s essential oil and bitter principles
underlie its medicinal and commercial significance (Wright, 2002).
A. absinthium extracts and essential oils are used
for healing various diseases (Lawless, 1999; Balz, 1996; Wright, 2002).
Oil of A. absinthium has been found to repel fleas and flies (Duke,
1995) and mosquitoes (Morton, 1981) and to kill houseflies (Kaul et
al., 1978). Anthelmintic, antibacterial, antifungal, insect repellent,
narcotic, digestive, tonic and other bioactivities are characteristic
of preparations from wormwood plants. Their stimulant property is dependent
on bitter substances as artabsin (sesquiterpene lactone) and absinthin
(dimmer of sesquiterpene lactones) present in plant extracts (Wright,
2002). Wormwood essential oil components 1, 8-cineole, cis (a) and trans
(ß)-thujones help people to withstand cold and other hardships of the
Himalayan region (Wright, 2002; Aslan et al., 2005; Kordali et
al., 2006). Large amounts of the above compounds are toxic (Woolf,
1999). Seizures may be caused by 1, 8- cineole and thujones for chronic
users. In addition, thujones may evoke dementia (Wright, 2002).
The major constituent of wormwood oil is thujone, present
at levels of approximately 40-70% of the oil (Tucker et al., 1993).
Other constituents present at significant levels include myrcene (<35%),
a- pinene (6%) and nerol (3%) in wormwood oil of Russian origin (Goraev
et al., 1962); camphor (6%), ?-cymene (4%), limonene (4%) and a-
pinene (4%) in Spanish wormwood oil (Mugica and Ochoa, 1974); ß-phellandrene
(10%), a-humachalene (7%) and ß- caryophyllene (5%) (Kaul et al.,
1979; Aslan et al., 2005; Kordali et al., 2006). High levels
of thujanol and thujyl acetate (60-70%), myrcene (35%), camphor and 1,
8-cineole were also determined in wormwood essential oils (Wright, 2002).
Thujones, trans-sabinyl acetate, cis-chrysanthenyl acetate and cis-epoxyocimene
are the most common constituents in wormwood essential oils (Arino et
al., 1999; Wright, 2002; Juteau et al., 2003).
This study was conducted to obtain information about essential
oil of wormwood growing wild in Iran. It presents the chemical composition
of essential oil from aerial parts of A. absinthium L. collected
in a habitat from North of Iran.
MATERIALS AND METHODS
Plant material: The aerial parts of A. absinthium were
collected in full blooming time from an area between Deylaman and Asiabar
villages, at Alborz altitudes in Guilan province in September 2005. The
aerial parts were dried at room temperature (20-25°C) for several days
until measurement of weight. Voucher specimens were deposited in the Herbarium
of the faculty of agricultural sciences, Shahed University, Tehran, Iran.
Isolation procedure: The essential oil was prepared by hydrodistillation
for 2.5 h using a Clevenger-type apparatus. The oil was dried over anhydrous
calcium chloride and stored in sealed vials at low temperature (2°C) before
Gas chromatography: GC analysis was performed using a Shimadzu
GC-9A gas chromatograph equipped with a DB-5 fused silica column (30 mx0.25
mm i.d., film thickness 0.25 µm). Oven temperature was held at 40°C for
5 min and then programmed to 250°C at a rate of 4°C min-1.
Injector and detector (FID) temperature were 260°C; helium was used as
carrier gas with a linear velocity of 32 cm sec-1.
Gas chromatography-mass spectrometry: GC-MS analyses were carried
out on a Varian 3400 GC-MS system equipped with a DB-5 fused silica column
(60 mx0.25 mm i.d.). Oven temperature was 40-250°C at a rate of 4°C min-1,
transfer line temperature 260°C, carrier gas helium with a linear velocity
of 31.5 cm sec-1, split ratio 1/60, ionization energy 70 eV,
scan time 1 sec, mass range 40-300 amu.
Identification of components: The components of the oil were identified
by comparison of their mass spectra with those of a computer library or
with authentic compounds and confirmed by comparison of their retention
indices, either with those of authentic compounds, or with data published
in the literature (Adams, 1995; Shibamoto, 1987).
RESULTS AND DISCUSSION
The chemical composition of the A. absinthium essential
oil is shown in Table 1. The components are listed in
order of their elution on the DB-5 column.
Chemical composition of the
essential oil of Artemisia absinthium
|RI: Retention index on DB-5 column
Twenty eight compounds were identified in the essential
oil of A. absinthium, representing 93.3% of the total oil components.
The major components of the oil were found to be ß- pinene and ß- thujone,
their contents being 23.8 and 18.6%, respectively. The other major components
were sabinene, Cubenol, linalool, myrcene, a-pinene, a- dehydro- ar-himachalene,
a-phellandrene and germacrene D. Monoterpenoids comprised a large part
(75.4%) of wormwood oil. The largest part of the essential oil was formed
by hydrocarbon monoterpenes (47.8%) and the other parts included oxygen-containing
monoterpenes (27.6%), hydrocarbon sesquiterpenes (9.1%) and oxygen-containing
sesquiterpenes (8.8%). Compounds with a sabinene carbon skeleton (sabinene,
a and ß- thujone, iso-3-thujanol) made up 29.3% of total oil.
The oil isolated by hydrodistillation from the aerial parts
of A. absinthium was found to be dark green to brown liquid that
can be attributed to the presence of chamazulene and its yield was 1.30%
(v/w). Essential oil contents between 0.2 and 1.5% in the crude drug have
been reported in the literature (Wichtl, 1989). The most abundant constituent
in the essential oil was ß-pinene, which has not been reported for the
species from other countries till now. It can be said that ß-pinene is
a new component for wormwood oil and possibly restricted to Iran. It has
been reported that A. absinthium plants produce several chemotypes
of essential oils in different countries and the main component of wormwood
essential oils is thujone (Juteau et al., 2003; Wright, 2002).
A sample collected in Argentina, had the ß-thujone (60%) as main constituent
(Sacco and Chialva, 1988). The plants and their parts from Croatia contained
notable amounts of (Z)-6, 7-epoxyocimene besides the first major constituent
trans-thujone (Juteau et al., 2003). Some volatile oils from North
Italy (Wright, 2002), France (Juteau et al., 2003; Wright, 2002)
and Spain (Arino et al., 1999) did not contain thujones. The predominant
components of the above oils were (Z)-6, 7-epoxyocimene and/or (Z)-chrysanthenyl
acetate. Two chemotypes were determined in Spain: one oil contained both
the above compounds as the main ones, but another oil was characterized
by (Z)-6, 7-epoxyocimene with minor amounts of other constituents (Arino
et al., 1999). Carnat et al. (1992) found cis- chrysanthenol
as the major component in the oil of plants grown in central France. Sabinyl
acetate prevailed in some oils from different countries (Arino et al.,
1999; Wright, 2002). Chiasson et al. (2001) reported ß-thujone
as a main component (32.1%) in wormwood essential oil from Canada. Oils
isolated from native and cultivated wormwood plants growing in Oregon
contained ß-thujone (40%) and cis-epoxyocimene (25%) in the native plants
and sabinene (30%) and myrcene (30%) in the cultivated plants (Tucker
et al., 1993). Bornyl acetate was reported as the major constituent
(23%) of a sample collected in Cuba (Pino et al., 1997), but it
was not observed in the present study.
It is suggested that the studied wormwood in this work synthesizes
a kind of essential oil component, which is different from other components
reported so far. The quality and yield of essential oils from Anthemideae
plants are influenced by the various factors such as harvesting season
(Cornu et al., 2001), fertilizer and the pH (ideal in acidic, pH
4.5-5.4) of soils (Alvarez-Castellanos and Pascual-Villalobos, 2003),
the choice and stage of drying conditions (Tateo and Riva, 1991), the
geographic location (Maffei et al., 1994), subspecies (Goren et
al., 2001), choice of plant part or genotype (Mishra et al.,
1999; Nori-Shargh et al., 1999; Keskitalo et al., 2001)
or extraction method (Scalia et al., 1999).
Monoterpenoids comprised a large part of wormwood oil in
the study. This corroborates the finding of Judpentiene and Mockute (2004)
which reported that monoterpenoids comprise a large part (55.7-80.2%)
of wormwood oil in Lithuania. The largest part of the essential oil was
formed by hydrocarbon monoterpenes (47.8%) and the other parts included
oxygen-containing monoterpenes (27.6%), hydrocarbon sesquiterpenes (9.1%)
and oxygen-containing sesquiterpenes (8.8%). Judpentiene and Mockute (2004)
reported that the largest part of the essential oil in wormwood is formed
by oxygenated monoterpenes (47.1-66.7%) and the hydrocarbon monoterpenes
Compounds with a sabinene carbon skeleton made up 29.3%
of total oil. These compounds were comprised (33.9-61.0%) of wormwood
oil in Lithuania (Judpentiene and Mockute, 2004). Chamazulene (0.9%) was
present in the oil which contributed to the characteristic colour of wormwood
oil. A small percentage of chamazulene (0.2%) was found in the Cuban oil
(Pino et al., 1997) and in two chemotypes studied of Spanish oil
it was 3 and 0.3% (Arino et al., 1999).
The authors gratefully acknowledge the financial support
of the University of Shahed and scientific assistance of Dr. H. Abbasipour.
1: Adams, R.P., 1995. Identification of Essential oil Components by Gas Chromatography/Mass Spectroscopy. 1st Edn., Allured Publishing Co., Illinois, USA., ISBN: 0-931710-42-1.
2: Alvarez-Castellanos, P.P. and M.J. Pascual-Villalobos, 2003. Effect of fertilizer on yield and composition of flowerhead essential oil of Chrysanthemum coronarium (Asteraceae) cultivated in Spain. Ind. Crop Prod., 17: 77-81.
Direct Link |
3: Arino, A., I. Arberas, G. Renobales, S. Arriaga and J.B. Dominguez, 1999. Essential oil of Artemisia absinthium from the Spanish Pyrenees. J. Essent. Oil Res., 11: 182-184.
4: Aslan, I., S. Kordali and O. Calmasur, 2005. Toxicity of vapours of Artemisia absinthium essential oils to the Tetranychus urticae Koch and Bemisia tabaci Genn. Fresenius Environ. Bull., 14: 413-417.
5: Balz, R., 1996. The Healing Power of Essential Oils. Lotus Light, USA.
6: Carnat, A.P., M. Madesclaire, O. Chavignon and J.L. Lamaison, 1992. Cis-Chrysanthenol, a main component in the essential oil of Artemisia absinthium L. growing in Auvergne (Massiv Central), France. J. Essent. Oil Res., 4: 487-490.
7: Chiasson, H., A. Belanger, N. Bostanian, C. Vincent and A. Poliquin, 2001. Acaricidal properties of Artemisia absinthium and Tanacetum vulgare (Asteraceae) essential oils obtained by three methods of extraction. J. Econ. Entomol., 94: 167-171.
CrossRef | Direct Link |
8: Cornu, A., A.P. Carnat, B. Martin, J.P. Coulon, J.L. Lamaison and J.L. Berdagué, 2001. Solid-phase micro-extraction of volatile components from natural grassland plants. J. Agric. Food Chem., 49: 203-209.
Direct Link |
9: Duke, J.A., 1985. CRC Handbook of Medicinal Herbs. 1st Edn., CRC Press, Inc., Florida, USA.
10: Goraev, M.I., U.S. Bazalitskaya and L.N. Lishtvanova, 1962. The terpene portion of the essential oil of Artemisia absinthium. Zeitschrift fur Naturforschung, 35: 2799-2802.
11: Goren, N., B. Demirci and K.H.C. Ba er, 2001. Composition of the essential oils of Tanacetum sp. from Turkey. Flavour Fragr. J., 16: 191-194.
12: Judpentienë, A. and D. Mockutë, 2004. Chemical composition of essential oils of Artemisia absinthium L. (wormwood) growing wild in Vilnius. Chemija, 15: 64-68.
Direct Link |
13: Juteau, F., I. Jerkovic, V. Masotti, M. Milos, J. Mastelic, J.M. Bessiere and J. Viano, 2003. Composition and antimicrobial activity of the essential oil of Artemisia absinthium from Croatia and France. Planta Med., 69: 158-161.
Direct Link |
14: Kaul, V.K., S.S. Nigam and A.K. Banerjee, 1978. Insecticidal activity of some essential oils. Indian J. Pharm., 40: 22-26.
15: Kaul, V.K., S.S. Nigam and A.K. Banerjee, 1979. Thin-layer and gas chromatographic studies of the essential oil of Artemisia absinthium L. Indian Perfumer., 23: 1-7.
16: Keskitalo, M., E. Pehu and J.E. Simon, 2001. Variation in volatile compounds from Tansy (Tanacetum vulgare L.) related to genetic and morphological differences of genotypes. Biochem. Syst. Ecol., 29: 267-285.
Direct Link |
17: Kordali, S., I. Aslan, O. Calmasur and A. Cakir, 2006. Toxicity of essential oils isolated from three Artemisia species and some of their major components to granary weevil, Sitophilus granarius (L.) (Coleoptera: Curculionidae). Ind. Crops Prod., 23: 162-170.
CrossRef | Direct Link |
18: Lawless, J., 1995. The Illustrated Encyclopedia of Essential Oils: The Complete Guide to the Use of Oils in Aromatherapy and Herbalism. Element Books Ltd., Shaftesbury, UK., ISBN-13: 978-1852307219, Pages: 256.
19: Maffei, M., M Mucciarelli and S. Scannerini, 1994. Essential oils from Achillea species of different geographic origin. Biochem. Syst.Ecol., 22: 679-687.
20: Mishra, D.K., S.N. Naik, V.K. Srivastava and R. Prasad, 1999. Effect of drying Matricaria chamomilla flowers on chemical composition of essential oil. J. Med. Aro. Plant Sci., 21: 1020-1025.
21: Morton, J.F., 1981. Atlas of Medicinal Plants of Middle America. Charles C. Thomas, Springfield, Illinois, USA., pp: 23-30.
22: Mugica, M.G. and J.T. Ochoa, 1974. Aceite esencial de Artemisia absinthium L. Contribucion al estudio 4e los aceites esenciales Espanoles II. Aceite esenciales de la provincia de Guadalajara. Instituto Nacionale Investigaciones Agrarias, Ministry of Agriculture, Madrid.
23: Nori-Shargh, D., H. Norouzi-Arasi, M. Mirza, K. Jaimand and S. Mohammadi, 1999. Chemical composition of the essential oil of Tanacetum polycephalum (sp. heterophyllum). Flavour Fragr. J., 14: 105-106.
24: Pino, J.A., A. Rosado and V. Fuentes, 1997. Chemical composition of the essential oil of Artemisia absinthium L. from Cuba. J. Essent. Oil Res., 9: 87-89.
25: Sacco, T. and F. Chialva, 1988. Chemical characteristics of the oil from Artemisia absinthium collected in Patagony (Argentina). Planta Med., 54: 93-97.
26: Scalia, S., L. Giuffreda and P. Pallado, 1999. Analytical and preparative supercritical fluid extraction of chamomile flowers and its comparison with conventional methods. J. Pharm. Biomed. Anal., 21: 549-558.
CrossRef | Direct Link |
27: Shibamoto, T., 1987. Retention Indices in Essential Oil Analysis. In: Capillary Gas Chromatography in Essential Oil Analysis, Sandra, P. and C. Bicchi (Eds.). Huethig-Verlag, New York, USA., pp: 259-274.
28: Tateo, F. and G. Riva, 1991. Influence of the drying process on the quality of essential oils in Artemisia absinthium. Mitt. Gebiete Levensm. Hyg., 82: 607-614.
29: Tucker, A.O. M.J. Maciarello and G. Sturtz, 1993. The essential oils of Artemisia Powis Castle and its putative parents A. absinthium and A. arborescens. J. Essent. Oil Res., 5: 239-242.
30: Wichtl, M., 1989. Teedrogen, 2. Auflage, Wissenschaftliche Verlagsgesellschaft mbH, Stuttgart.
31: Woolf, A., 1999. Essential oil poisoning. J. Toxicol., 37: 721-727.
32: Wright, C.W., 2002. Artemisia. CRC Press, Boca Raton, FL., USA., ISBN-13: 978-0415272124, Pages: 344.
33: Zargari, A., 1989. Medicinal Plants. University of Tehran Press, Tehran.