RESEARCH PAPER
Essential oil composition and in vitro biological activity of Achillea millefolium L. extracts
 
More details
Hide details
1
Department of Virology and Immunology, Institute of Microbiology and Biotechnology, Maria Curie-Skłodowska University, Lublin, Poland
2
Department of Chemistry, Laboratory of Planar Chromatography, Medical University, Lublin, Poland
3
Department of Surgery, District Hospital, Lublin, Poland
CORRESPONDING AUTHOR
Roman Paduch   

Maria Curie-Skłodowska University, Institute of Microbiology and Biotechnology, Department of Virology and Immunology, Akademicka 19, 20-033 Lublin, Poland.
 
J Pre Clin Clin Res. 2008;2(1):49–58
KEYWORDS
ABSTRACT
The Achillea genus is one of the widely used genera to treat various medical ailments. In this study, gas chromatography (GS) and Solid Phase Microextraction (SPME) were used to determine the essential oil composition of the A. millefolium L. Human skin fi broblasts (HSF) viability based on spectrophotometrical 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Neutral Red (NR) methods and morphological analysis was performed in in vitro cell culture. Free radical scavenging activity of ethanol, ethyl acetate and water extracts of A. millefolium L. was also measured. The total oil content in Achillea fl owers was analyzed only with gas chromatography/mass spectrometry (GC/MS) method. SPME-GC/MS technique revealed that Achillea fl owers liberate high amounts of monoterpenes (α-pinene; β-pinene; 1,8-cineole). However, proportionally, most of all sesquiterpenes (β-caryophyllene and germacrene D) were found. We also showed that fresh fl owers contain much more monoterpenes such as α-pinene or sabinene but fewer amounts of β-pinene than preserved material. MTT analysis indicated that methanol extract from fresh, not dried, plant material and water extract from fresh plants at doses ranging from 25-225 μg/mL had no cytotoxic activity on HSF cells. Methanol extract from dried and freeze herb (doses >75 μg/mL) and ethyl acetate extract from fresh plants (doses >25 μg/mL) inhibited succinyl dehydrogenase activity. In the NR assay, only methanol extract from fresh plant material had no cytotoxic activity. The remainder signifi cantly decreased dye uptake. Moreover, ethyl acetate extract strongly infl uenced cellular actin fi laments composition, and consequently morphology of cells. Analysis of the IC50 values revealed that for methanol from fresh herb and water extracts, the IC50 values are higher than 250 μg/mL. The lowest IC50 was 13 μg/mL, obtained in NR assay for ethyl acetate extract. The highest free radical scavenging activity (1,1-diphenyl-2-picrylhydrazyl (DPPH) method) was found for methanol extract from dried herb at the maximal dose used (175 μg/mL), and was 19.2% higher than control value. When compared to the Trolox standard activity it was equivalent of the 6.8 μg/mL. In conclusion, the results suggest that A. millefolium L. extracts may, in limited concentrations, be useful in preparations of topical application and considered for use in clinical dermatology.
 
REFERENCES (21)
1.
Dokhani S, Cottrell T, Khajeddin J, Mazza G: Analysis of aroma and phenolic components of selected Achillea species. Plant Foods Hum Nutr 2005, 60, 55-62.
 
2.
Candan F, Unlu M, Tepe B, Daferera D, Polissiou M, Sokmen A, Akpulat A: Antioxidant and antimicrobial activity of the essential oil and methanol extracts of Achillea millefolium subsp. Millefolium Afan. (Asteraceae). J Ethnopharmacol 2003, 87, 215-220.
 
3.
Innocenti G, Vegeto E, Dall’Acqua S, Ciana P, Giorgetti M, Agradi E, Sozzi A, Fico, G, Tome F: In vitro estrogenic activity of Achillea millefolium L. Phytomedicine 2007, 14, 147-152.
 
4.
Cavalcanti AM, Baggio CH, Freitas CS, Rieck L, de Sousa RS, Da Silva- Santos JE, Mesia-Vela S, Marques MCA: Safety and antiulcer effi cacy studies of Achillea millefolium L. after chronic treatment in Wistar rats. J Ethnopharmacol 2006, 107, 277-284.
 
5.
Salvagnini LE, Migliato KF, Isaac VLB, Correa MA, Salgado HRN, Pietro RCLR: Evaluation of effi cacy of preservatives associated with Achillea millefolium L. extracts against Bacillus subtilis. Braz J Microbiol 2006, 37, 75-77.
 
6.
Boskovic Z, Radulovic N, Stojanovic G: Essential oil composition of four Achillea species from the Balkans and its chemotaxonomic signifi cance. Chem Nat Compounds 2005, 41, 674-678.
 
7.
Gulcin İ: Antioxidant activity of caff eic acid (3,4-dihydroxycinnamic acid). Toxicology 2006, 217, 213-220.
 
8.
Dijoux N, Guingand Y, Bourgeois C, Durand S, Fromageot C, Combe C, Ferret P-J: Assessment of the phototoxic hazard of some essential oils using modifi ed 3T3 neutral red uptake assay. Toxicol in Vitro 2006, 20, 480-489.
 
9.
Paduch R, Kandefer-Szerszeń M, Trytek M, Fiedurek J: Terpenes: substances useful in human healthcare. Arch Immun Ther Exp 2007, 55, 315-327.
 
10.
Hayes AJ, Markovic B: Toxicity of Australian essential oil Backhousia citriodora (lemon myrtle). Part 2. Absorption and histopathology following application to human skin. Food Chem Toxicol 2003, 41, 1409-1416.
 
11.
Lima CF, Carvalho F, Fernandes E, Bastos ML, Santos-Gomes PC, Fernandes-Ferreira M, Pereira-Wilson C: Evaluation of toxic/protective eff ects of the essential oil of Salvia offi cinalis on freshly isolated rat hepatocytes. Toxicol in Vitro 2004, 18, 457-465.
 
12.
Fabian D, Sabol M, Domaracka K, Bujňakova D: Essential oils – their antimicrobial activity against Escherichia coli and eff ect on intestinal cell viability. Toxicol in Vitro 2006, 20, 1435-1445.
 
13.
Soekanto A, Kasugai S, Mataki S, Ohya K, Ogura H: Toxicity of camphorated phenol and camphorated parachlorophenol in dental pulp cell culture. J Endod 1996, 22, 284-286.
 
14.
Bhaya M, Beniwal R: Camphor induced myocarditis: A case report. Cardiovasc Toxicol 2007, 7, 212-214.
 
15.
Ferreira A, Proenca C, Serralheiro MLM, Araujo MEM: The in vitro screening for acetylcholinesterase inhibition and antioxidant activity of medicinal plants from Portugal. J Ethnopharmacol 2006, 108, 31-37.
 
16.
Lourens ACU, Reddy D, Başer KHC, Viljoen AM, Van Vuuren SF: In vitro activity and essential oil composition of four indigenous South African Helichrysum species. J Ethnopharmacol 2004, 95, 253-258.
 
17.
Şahin F, Gulluce M, Daferera D, Sokmen A, Sokmen M, Polissiou M, Agar G, Ozer H: Biological activities of the essential oils and methanol extracts of Origanum vulgare ssp. Vulgare in the Eastern Anatolia Region of Turkey. Food Control 2004, 15, 549-557.
 
18.
Farhat GN, Aff ara NI, Gali-Muhtasib HU: Seasonal changes in the composition of the essential oil extract of East Mediterranean sage (Salvia libanotica) and its toxicity in mice. Toxicon 2001, 39, 1601-1605.
 
19.
Chudnicka A, Matysik G: Research of enzymatic activities of fresh juice and water infusions from dry herbs. J Ethnopharmacol 2005, 99, 281-286.
 
20.
Ozbek H, Uğraş S, Dulger H, Bayram İ, Tuncer İ, Ozturk G, Ozturk A: Hepatoprotective eff ect of Foeniculum vulgare essential oil. Fitoterapia 2003, 74, 317-319.
 
21.
De-Oliveira ACAX, Ribeiro-Pinto LF, Paumagartten FJR: In vitro inhibition of CYP2B1 monooxygenase by β-myrcene and other monoterpenoid compounds. Toxicol Lett 1997, 92, 39-46.
 
eISSN:1898-7516
ISSN:1898-2395