Toxicity tests of wild wormwood essential oils were performed using the brine shrimp (Artemia sp.) test. N auplii lethality (LC 50) ranged 15.7-31.9 µg/mL, depending on oil composition. The most toxic A. absinthium oils were found to be those containing appreciable amount of trans-sabinyl acetate (45.2%) and (cis+trans) thujones (12.3%), while other samples with equivalent amounts of sabinyl acetate, but without thujones were determined to be notably less toxic. Herb material for the tests was collected in Lithuania, the volatile oils were obtained by hydrodistillation from different plant organs (inflorescences and leaves) and analysed by GC-MS

Asta Judzentiene

Isabelle Laffont-Schwob

Jurga Budiene

Renata Gircyte

Véronique Masotti

International audienceToxicity tests of wild wormwood essential oils were performed using the brine shrimp (Artemia sp.) test. Nauplii lethality (LC 50) ranged 15.7-31.9 µg/mL, depending on oil composition. The most toxic A. absinthium oils were found to be those containing appreciable amount of trans-sabinyl acetate (45.2%) and (cis+trans) thujones (12.3%), while other samples with equivalent amounts of sabinyl acetate, but without thujones were determined to be notably less toxic. Herb material for the tests was collected in Lithuania, the volatile oils were obtained by hydrodistillation from different plant organs (inflorescences and leaves) and analysed by GC-MS

Judzentiene, Asta

Budiene, Jurga

Gircyte, Renata

Masotti, Veronique

Laffont-Schwob, Isabelle

HAL AMU

Toxic Activity and Chemical Composition of Lithuanian Wormwood (Artemisia absinthium L.) Essential Oils

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Toxic Activity and Chemical Composition of Lithuanian
Wormwood (Artemisia absinthium L.) Essential Oils
Asta Judzentiene, Jurga Budiene, Renata Gircyte, Veronique Masotti, Isabelle
Laffont-Schwob
To cite this version:
Asta Judzentiene, Jurga Budiene, Renata Gircyte, Veronique Masotti, Isabelle Laffont-Schwob. Toxic
Activity and Chemical Composition of Lithuanian Wormwood (Artemisia absinthium L.) Essential
Oils. Records of Natural Products, ACG Publications, 2012, 6, pp.180 - 183. ￿hal-02095676￿
SHORT REPORT                                                                                     
 
The article was published by Academy of Chemistry of Globe Publications 
www.acgpubs.org/RNP © Published  10/23/2011 EISSN: 1307-6167 
 
 
 
 
Rec. Nat. Prod. 6:2 (2012) 180-183 
 
 
Toxic Activity and Chemical Composition of Lithuanian 
Wormwood (Artemisia absinthium L.) Essential Oils  
 
Asta Judzentiene
1*
, Jurga Budiene
1
, Renata Gircyte
1§
, Véronique Masotti
2
 
and Isabelle Laffont-Schwob
2
 
 
1 Center for Physical Sciences and Technology, Institute of Chemistry, A. Gostauto 9, LT-01108, 
Vilnius, Lithuania 
 
2 University of Provence, Institut Méditerranéen d’Écologie et de Paléoécologie, UMR-IMEP-CNRS-
IRD 6116, Équipe BBE, Case 97, 3 place Victor Hugo, 13331 Marseille Cedex 3, France 
 
 
 
 (Received March 28, 2011; Revised July 19, 2011; Accepted July 24, 2011) 
 
Abstract: Toxicity tests of wild wormwood essential oils were performed using the brine shrimp (Artemia sp.) 
test. Nauplii lethality (LC50) ranged 15.7-31.9 µg/mL, depending on oil composition. The most toxic A. 
absinthium oils were found to be those containing appreciable amount of trans-sabinyl acetate (45.2%) and 
(cis+trans) thujones (12.3%), while other samples with equivalent amounts of sabinyl acetate, but without 
thujones were determined to be notably less toxic. Herb material for the tests was collected in Lithuania, the 
volatile oils were obtained by hydrodistillation from different plant organs (inflorescences and leaves) and 
analysed by GC-MS. 
 
Keywords: Artemisia absinthium; essential oil composition; GC-MS; trans-sabinyl acetate; (cis-trans) thujones; 
(Z)-epoxy-ocimene; brine shrimp lethality. 
 
1. Plant Source 
 Artemisia absinthium (wormwood, Asteraceae) is used as a medicinal remedy worldwide. The 
wormwood extracts possess numerous properties, such as antiprotozoal, cytotoxic, antimalarial, 
antipyretic, antiradical and antioxidant, antimicrobial, anthelmintic, antidepressant, neurotropic etc. 
                                               
* Corresponding author: E-Mail: judzent@ktl.mii.lt; Phone: +370 5 264-8841; Fax: +370 5 264-9774.  
§ permanent address: Vilnius University, Faculty of Chemistry, Naugarduko 24, LT-03225, Vilnius, Lithuania 
 
 
Judzentiene et al., Rec. Nat. Prod. (2012) 6:2 180-183  
 
 
181
This plant is widely used in the Lithuanian folk medicine mainly for stomachic and anthelmintic 
activity, also for healing gall-bladder and kidneys diseases, against insomnia.  
 
2. Previous Studies 
 
A long list of various biological activities, such as acaricidal, antifungal, antibacterial, 
antimicrobial, neurotropic etc. has been reported for wormwood essential oils [1-7]. Meanwhile, data 
on toxicity of wormwood oils are limited, so the aim of our study is to evaluate toxic activity of A. 
absinthium essential oils linked to the natural variations of composition of the oils from plants 
growing wild.  
 
3. Present Study 
 
 The aerial parts of individuals from Artemisia absinthium L. populations were collected at full 
flowering stage (in August, 2008) in Western and South parts of Lithuania: A - Klaipeda district and B 
- Druskininkai, Latezeris. Voucher specimens were deposited in the herbarium (Institute of Botany 
(BILAS), Vilnius) with numbers: A-68929 and B-68930. The essential oils were obtained by 
hydrodistillation for 2 h using a Clevenger-type apparatus. Oils of dark orange-brown color were 
obtained and their yields ranged from 1.9 to 2.3% (v/w) on a dry weight basis. 
 GC/MS: Analyses by GC/MS were performed using a chromatograph HP 5890 interfaced to 
an HP 5971 mass spectrometer (ionization voltage 70 eV, scan time 0.6 s, scan range 35-400 Da) and 
equipped with a capillary column DB-5 (50 m × 0.32 mm i. d., film thickness 0.25 µm). The oven 
temperature was held at 60°C for 2 min, then programmed from 60 to 160°C at a rate 5°C/min,
 
held 
for 1 min, then heated from 160 °C to 250 °C at the rate 10°C/min and finally isothermal at 250°C for 
3 min, using He as a carrier gas (1.0 mL/min), split 1:40. Injector and detector temperatures were 
250°C. Qualitative analysis was based on a comparison of retention times, indexes and mass spectra 
with the corresponding data in the literature [8], co-injection of some terpenes references and 
computer mass spectra libraries (Wiley and NBS 54K).  
 GC/FID: An HP 5890 gas chromatograph, equipped with a FFAP column (30m × 0.25mm; 
0.25µm) was used for quantitative analysis. The GC oven was programming from 70°C (10 min) to 
250°C at a rate 3°C/min, using He as a carrier gas (0.7 mL/min). The injector and detector 
temperatures were 230 and 250°C, respectively.  
Data on chemical composition of the different organs i.e. inflorescences and leaves of 
wormwood essential oils are presented in Table 1. Twenty-eight identified components (additionally, 
three unknown compounds) constituted up to 79.8-94.1% of total oil content. Monoterpenoids fraction 
was the major group of compounds in both inflorescence and leaf oils, comprising 55.2-84.7% of the 
oils. Amounts of oxygenated monoterpenes were higher in the inflorescence oils (≤70.7%) than that in 
leaves (≤60.9%). Compounds with sabinane skeleton were characteristic of all the oils. Trans-sabinyl 
acetate was the main constituent of all the oils. Thujones and (Z)-epoxy-ocimene were the second 
main compounds for Klaipeda and Druskininkai populations, respectively. The oils could be attributed 
to two different chemotypes: sabinyl acetate + thujones and sabinyl acetate + (Z)-epoxy-ocimene.  
Toxicity test: Toxic activity of the wormwood oils was tested in vivo, using brine shrimp 
Artemia sp. (larvae) [9]. The eggs of shrimps hatch within 48 hours to provide larvae (nauplii) in sea 
water (31g/L sea salt) at 20-25°C. Then, different amounts of wormwood essential oils dissolved in 
dimethyl sulfoxide (DMSO) were added and survivors were counted after 24 hours. Lethality (LC50) of 
nauplii was calculated (n=4, with 95% confidence interval). Control test was done with DMSO.  
Test of wormwood essential oil toxicity has shown that lethality (LC50) of brine shrimp larvae 
ranged between 15.7-31.9 µg/mL. Differences in the level of toxicity could be explained by the 
various quantities of main toxic constituents. The most toxic oil was defined as containing, in addition 
to the major constituents sabinyl acetate (45.2%) and thujones (12.3%), also appreciable amounts of 
myrcene (6.0%) and 1,8-cineole (5.3%).  
 
Essential oil of Artemisia absinthium  
 
182
 
Table 1. Main composition (over 1.0%) of inflorescence and leaf essential oils of A. absinthium from 
Lithuania 
RI Compounds AF AL BF BL 
975 Sabinene 6.5 5.5 1.7 1.6 
989 β-Pinene 1.5 3.5 2.0 3.0 
990 Myrcene 6.0 2.0 4.6 3.8 
1002 α-Phellandrene t t  t 
1031 1,8-Cineole 5.3 4.9   
1037 (Z)-β-Ocimene t t 1.3 1.1 
1096 Linalool 2.0 2.2 2.0 1.6 
1102 cis-Thujone 7.0 6.5 t  
1114 trans-Thujone 5.3 4.6 t  
 (cis+trans)-Thujones 12.3 11.1 t   
1132 (Z)-Epoxy-ocimene 2.9 3.5 12.8 8.5 
1142 trans-Sabinol 0.5 0.4 1.5 1.4 
1164 cis-Chrysanthenol  t   
1177 Terpinen-4-ol 2.5 1.3 t  
1195 Myrtenol   1.6 1.2 
1290 trans-Sabinyl acetate 45.2 37.5 40.5 33.0 
1419 trans-Caryophyllene 1.5 1.7 1.8 2.3 
1470 Acoradiene 1.0 2.0 0.9 1.2 
1477 Geranyl propionate 0.3 0.5 1.7 2.0 
1485 Germacrene D 0.2 0.6 1.9 2.6 
1506 Unknown 1   0.4 2.9 
1509 Lavandulyl isovalerate 1.1 0.8 2.4 2.3 
1511 Lavandulyl 2-methyl butyrate  1.0 0.6 2.1 0.7 
1515 Geranyl isobutanoate 1.1 0.4 0.4 0.5 
1564 Geranyl butyrate   0.9  
1563 trans-Nerolidol 0.2   1.1 
1583 Caryophyllene oxide 0.3  0.9 1.2 
1605 Unknown 2 0.8 0.6 2.1 3.0 
1615 Unknown 3  0.3 0.4 3.1 3.6 
1731 Chamazulene 0.3 0.3 0.4 0.9 
1756 Nuciferol acetate  0.5 1.2 1.8  
1860 Lanceol acetate  0.8 1.5 2.8 0.3 
 Total  94.1  82.5  91.6  79.8  
 Oxygenated monoterpenes  70.7  60.9  58.4  45.7   
 Monoterpene hydrocarbons 14.0  11.0  9.6  9.5   
 Monoterpenoids 84.7  71.9  68.0  55.2   
 Compounds with sabinane skeleton 57.5  54.5  43. 7  36.0   
      
RI-Retention indices on nonpolar column DB-5, t:- traces (≤0.05%). Inflorescences indicated by letter F, leaves - 
by L, growing localities indicated as follow: A - Klaipeda (Western Lithuania) and B - Druskininkai, Latezeris 
(South Lithuania).  
Mass spectra of unknowns identified tentatively: 
1: (M
+
 187): 157(100); 142(50); 186(43); 171(29); 141(29); 143(23); 128(19); 115(16); 156(16); 77(6); 187(6); 
91(5); 51(3); 39(3); 102(1). 
2: (M+ 187): 157(100); 186(77); 143(60); 142(59); 128(47); 171(45); 141(45); 115(33); 129(32); 156(21); 
77(12); 91(12); 187(11); 39(5); 41(5); 63(5); 105(4). 
3: (M
+
 187): 171(100); 186(81); 143(64); 129(51); 128(49); 157(41); 142(40); 156(35); 115(34); 141(33); 
91(25); 77(23); 79(16); 144(15); 172(15); 187(15); 93(13); 39(11); 41(11); 105(10); 65(9). 
 
Since compounds present in the greatest proportions are mostly responsible for the greatest share of 
the total toxicity, the involvement of the less abundant constituents should also be considered. It is 
well known that thujone is a very toxic (mostly neurotoxic) compound, while sabinyl acetate is 
fetotoxic, a skin irritant and an abortifacient [10]. Other tested samples with equivalent amounts of 
sabinyl acetate but with appreciable amounts of (Z)-epoxy-ocimene (≤12.8%) and without thujones 
Judzentiene et al., Rec. Nat. Prod. (2012) 6:2 180-183  
 
 
183
were found to be notably less toxic. Previously, it was evaluated that wormwood oils rich in (Z)-
epoxy-ocimene have antimicrobial properties [3]. 
 
Table 2. Toxic activity of A. absinthium essential oils against Artemia sp. nauplii 
Toxic activity value AF AL BF BL 
LC50 , µg/mL 15.7 18.6 25.8 31.9 
LC95 , µg/mL 44.2 56.9 91.2 150.3 
Acknowledgements  
The work has been performed in “Gilibert” programme and supported by the Lithuanian State 
Science and Studies Foundation (No.V-05/2007 (V-07025) and V-08/2008 (V-08033)). R. Gircyte’s 
participation in this research was supported by the Lithuanian Science Council Student Research 
Fellowship Award (No. SP-03/SP 08-94). 
Furthermore, A. Judzentiene is grateful to the Embassy of France in Lithuania for a French 
government grant (EGIDE, No.484214F). 
 
References 
[1]  K. Karwowska, J. Stegman, W. Duszkiewicz-Reinhard and A. Dobrzeniecka (1997). Studies on isolation 
and chemical composition of biologically active compounds of wormwood (Artemisia absinthium), Ann. 
Warsaw Agricult. Univ. – SGGW Horticult. 18, 105-108.  
[2]  H. Chiasson, A. Bélanger, N. J. 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. 94:1, 167-171. 
[3]  F. Juteau, I. Jerkovic, V. Masotti, M. Milos, J. Mastelic, J. M. Bessiére and J. Viano (2003). Composition 
and antimicrobial activity of the essential oil of Artemisia absinthium from Croatia and France, Planta 
Med. 69, 158-161.  
[4]  S. Kordali, A. Cakir, A. Mavi, H. Kilic and A. Yildirim (2005). Screening of chemical composition and 
antifungal and antioxidant activities of the essential oils from three Turkish Artemisia species, J. Agric. 
Food Chem. 53:5, 1408-1416. 
[5]  S. Kordali, R. Kotan, A. Mavi, A. Cakir, A. Ala and A. Yildirim (2005). Determination of the chemical 
composition and antioxidant activity of the essential oil of Artemisia dracunculus and of the antifungal and 
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spicigera essential oils, J. Agric. Food Chem. 53:24, 9452-9458. 
[6]  S. Kordali, I. Aslan, O. Çalmaşur 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.  
[7]  D. Lopes-Lutz, D. S. Alviano, C. S. Alviano and P. P. Kolodziejczyk (2008). Screening of chemical 
composition, antimicrobial and antioxidant activities of Artemisia essential oils, Phytochemistry 69:8, 
1732-1738. 
[8]  R. P. Adams (2001). Identification of essential oil components by gas chromatography/quadrupole mass 
spectroscopy. Allured publishing Co, Carol Stream, Illinois.  
[9]  J. L. McLaughlin, L. L. Rogers and J. E. Anderson (1998). The use of biological assays to evaluate 
botanicals, Drug Inform. J. 32, 513-524. 
[10] N. Pages, G. Fournier, C. Baduel, N. Tur and M. Rusnac (1999). Sabinyl acetate, the main component of 
Juniperus sabina l’Hérit. essential oil, is responsible for antiimplantation effect, Phytother. Res. 10:5, 438-
440. 
 
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Toxic Activity and Chemical Composition of Lithuanian Wormwood (Artemisiaabsinthium L.) Essential Oils

Toxic Activity and Chemical Composition of Lithuanian Wormwood (Artemisiaabsinthium L.) Essential Oils

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