Antibacterial Activityand Comparison of the Volatile Oils of Tanacetum tenuisectum (Boiss.) Podl. Obtained by Three Different Methods of Extraction

authors:

avatar Shiva Masoudi , avatar Jila Abbassi

Department of Chemistry, Islamic Azad University, Central Tehran Branch, Tehran, Iran.
Warning: No corresponding author defined!
Iranian Journal of Pharmaceutical Research : IJPR: Vol.16, issue Suppl; 188-196
article type: Original Article
received: December , 2015
accepted: May , 2016
DOI: https://doi.org/10.22037/ijpr.2017.2008

how to cite: Masoudi S, Abbassi J. Antibacterial Activityand Comparison of the Volatile Oils of Tanacetum tenuisectum (Boiss.) Podl. Obtained by Three Different Methods of Extraction. Iran J Pharm Res. 2017;16(Suppl):e124917. https://doi.org/10.22037/ijpr.2017.2008.

Abstract

The essential oils obtained by hydrodistillation (HD), steam distillation (SD) and solvent free microwave extraction (SFME) from the stems and flowers of Tanacetumtenuisectum (Boiss.) Podl., which is endemic to Iran, were analyzed by combination of GC and GC/MS.
Camphor (26.91 %, 27.23% and 25.52%), borneol (12.61%,11.48% and 7.62%) and 1,8-cineole (7.93%, 13.23% and 11.26%) were the main constituents of the HD,SD and SFME oils of Tanacetumtenuisectum respectively.
All three oils were rich in regard to monoterpenes and small percentage of sesquiterpenes and non terpenoid compounds.
Antibacterial activity of the essential oil of the plant was determined against six Gram positive and Gram negative bacteria. The results showed that this oil was active against all of the tested bacteria.

Introduction

The genus Tanacetum, which is an important member of the Compositaefamily, is widespread in Europe and Western Asia and consists about 150-200 species.The flora of Iran comprises 26 species of Tanacetumofwhich 12 are endemic (1, 2).

Some members of this genus have traditionally been used as a spicy additive for food, in cosmetics and as herbal remedies due to their biologically active compounds (3). Especially Tanacetumpartheniumhasbeenused sinceancienttimes for a variety of medicine proposes, and recently has gained considerableprominence due to its ability of alleviating the symptoms of migraine, athritisand psoriasis, and to inhibition of blood platelet aggregation (4).

According to recent studies, essential oils and extracts of members of the genus Tanacetumexhibit anti-inflammatory (5-7) anticancer (8) antibacterial (9, 10) antiviral (11) antifungal (12) insecticidal (13) and antiprotozoal effects (14, 15).

This genus is also found to contain sesquiterpene lactones (16, 17) a large group of molecules with several biological activities (18, 21).

Previous chemical investigations on different Iranian species ofTanacetum have shown that they possess sesquiterpene lactones (22, 23) and essential oils (24-32).

As part of our ongoing work on the chemical analysis of volatile obtained from wild plants of Iran, we report the composition of the volatile oils of Tanacetumtenuisectum obtained by hydrodistillation, steam distillation and solvent free microwave extraction with its antibacterial activity.

Experimental

Plant material

The stems and flowers of Tanacetumtenuisectum (syn. Pyrethrum tenuisectumBoiss., Chrysanthemum gaubaeBornm. and Chrysanthemum tenuisectum (Boiss.) Parsa), which is endemic to Iran, was collected from Rameh area, Garmsar, Province of Semnan, Iran in July 2013, during the flowering stage.

Voucher specimens have been deposited at the Herbarium of the Research Institute of Forests and Rangelands (TARI), Tehran, Iran.

Isolation of the essential oils

Distillation

Air-dried ground stems and flowers of the plant were separately subjected

tohydrodistillation and steam distillation using a Clevenger type apparatus for 3h. The obtained essential oils were dried over anhydrous sodium sulfate and after filtration, stored at 4ºC until tested and analyzed. The yield was found to be 0.2% and 0.3% (w/w), respectively.

Solvent Free microwave extraction

SFME extractionwas performed in a Milestone ETHOS 1600 batch reactor, which is a multimode microwave reactor operating at 2455 MHZ with a maximum delivered power of 1000 W, variable in 10 W increments. The dimensions of the PTFE-coated cavity are 35×35×35 cm. During the experiment time, temperature, pressure, and power were controlled using the "easy-WAVE" software package. Temperature was monitored with the aid of a shielded thermocouple (ATC-300) inserted directly in to the sample container.

Ina typical SFMEprocedure, 250 g of dry stems and flowers of T.tenuisectum were moistened prior to extraction by soaking in water for 1 h, then draining off the excess water.This step is essential to give thestems and flowers the initial moisture. Moistened stems and flowers were next placed in a reactor without any added solvent or water. The essential oil is collected, dried with anhydrous sodium sulfate and stored at 0ºC until used.

Gas chromatography

GC analysis was performed on Schimadzu15 A gas chromatograph equipped with a split/splitless injector (25ºC) and a flame ionization detector (250ºC). Nitrogen was used as carrier gas (1 mL/min) and the capillary column used was DB-5 (50m×0.2mm, film thickness 0.32µm).The column temperature was kept at 60ºC for 3 min and then heated to 220ºC witha 5ºC/ min rate and kept constant at 220ºC for 5 min. Relative percentage amount were calculated from peakarea usinga SchimadzuC-R4 Achromatopac without the use of correction factors.

Gas chromatography - mass spectrometry

Analysis was performed using a Hewlett- Packard 5973 with a HP-5MS column (30m×0.25 mm, film thickness 0.25 µm). The column temperature waskept at 60ºC for 3 min and programmed to 220ºC at a rate of 5ºC/min and kept constant at 220ºC/min for 5 min.The flow rate of Helium as carrier gas with (1 mL/min).

Mass spectrometry wastaken at 70 eV. The retention indices for all the components were determined according to the Van Den Door method, using n- alkanes as standards (33).

The compounds were identified by (RRI, DB5) with those reported in the literature and by comparison of their mass spectra with the Wiley library or with the published mass spectra (34, 35).

Antibacterial assay

The antibacterial activity of the essential oil from the aerial parts of Tanacetumtenuisectum was evaluated by disc diffusion method using Mueller- Hinton Agar (36). The antibacterial activity of the essential oil of the plant was tested against three Gram- positive and three Gram-negative bacteria.

The Gram-positive bacteria included Staphylococcus aureusATCC 25923, Bacillus subtilis ATCC 9372, and Bacillus creus ATCC 6633, and the Gram-negative bacteriaincluded Klebsiellapneumoniae ATCC 27736, Entobacteraerogenes ATCC 49469, and Escherichia coil ATCC 25922. The bacteria were obtained from the Iranian Research Organization of Science and Technology.

A serial dilution of the oil was prepared in Mueller-Hinton Broth forbacteria. The oil was diluted by the water and ethanol solvents. The solvents, at an appropriate concentration were also used as a negative centrol. The standardized suspension of bacteria was incubated in to each tube. The tubes were incubated at37ºCfor 24h.Thelowestoil concentration, where there was no visible growth, was the Minimum Inhibitory Concentration (MIC) when compared to control.

To determine the Minimum Bactericidal Concentration (MBC), for each set of test tubes in the MIC determination, a loopful of broth was collected from those tubes which did not show any growth and incubated on Muller- Hinton Agar by streaking. Plates incubated with bacterial were then incubated at 37ºC for 24 h. After incubation, the concentration at which no visible growth was seen was noted as MBC for bacteria. All the experiments were carried out in triplicate and mean calculated.

Results and Discussion

The identified volatile components and their peak area percentages of the stems and flowers of Tanacetumtenuisectumobtained by hydrodistilliation, steam distillation and solvent free microwave extraction are given in Table 1. The components are listed in order of their elution on the DB-5 column.

As it is shown from the Table 1, about 96.87% (34 components) of the hydrodistilled oil, 91.32% (46 constituents) of the steam distilled oil and 95.1% (44 components) of the solvent free microwave extraction oil of T.tenuisectumwere identified. The main components in three oils were camphor (26.91%, 27.23% and 25.52%), borneol (12.61%, 11.48% and 7.62%) and 1,8- cineole (7.93%, 13.23% and 11.26%), respectively. Other notable constituents were in hydrodistilled oil hexadecanoic acid (7.30%), carotol (6.58%) and γ-eudesmol (5.75%); in steam distilled oil camphene (5.44%), hinesol (4.35%) and (E)-sequilavandulol (4.24%) and in solvent free microwave extraction oil hinesol (6.86%), hexadecanoic acid (6.55%) and (E) - sequilavandulol (4.89%).

Table 1

Comparative chemical composition (%) of Tanacetumtenuisectum oil obtained by HD, SD and SFME.

No.CompoundsaRIbHD(%)SD(%)SFME(%)
1Hexanal800-0.24-
2Tricyclene924-0.37-
3α-Thujene928-0.19t
4α- Pinene935-1.511.00
5Camphene9510.705.443.92
6β- Pinene981-1.040.68
7Mesitylene994-0.33-
8p- Cymene1024t1.841.17
91,8-Cineole10337.9313.2311.26
10γ-Terpinene1062-0.42t
11Linalool10960.730.62t
12Camphor114126.9127.2325.52
13cis-Chrysanthenol11600.82-1.09
14Pinocarvone1162-0.32-
15Borneol116512.6111.487.62
16Terpin-4-ol11771.140.990.59
17Naphthalene1183-0.25-
18α-Terpineol11881.200.840.58
19Myrtenal1191-0.22-
20Myrtenol1193-0.24-
21trans-Carveol12170.640.780.54
22cis-Chrysanthenyl acetate1260t0.23t
23Bornyl acetate12851.661.401.12
242- methyl Naphthalene1292-1.000.86
251- methyl Naphthalene1306-0.63t
26α-Terpinyl acetate1350-0.3-
No.CompoundsaRIbHD(%)SD(%)SFME(%)
27Decanoic acid13522.03-0.19
28α-Copaene1376-0.27-
292,6-dimethyl Naphthalene1379--0.57
30(E)-β-Damascenone1380-t-
311,7- dimethyl Naphthalene1397--0.44
322,7- dimethyl Naphthalene1400-0.54-
331,3- dimethyl Naphthalene1415-0.46-
34β-Caryophyllene1418--0.73
35Neryl acetone14310.80-0.88
36isobutyl-n-Butyrate14711.12--
37Pentadecane1500-0.17-
38β-Bisabolene1509--0.56
39Elemol1546--0.41
40(E)- Nerolidol15641.070.361.37
41(z)-dehydroApofarnesol15680.70-0.77
42Spathulenol15742.751.111.37
43Caryophyllene oxide15813.431.402.01
44Carotol15946.58--
45Tetradecanal16101.30-1.23
46(E)- isoEugenol acetate1611-0.39-
47γ-Eudesmol16305.75--
48(E)- Sesquilavandulol1632-4.244.89
49β- Caryophylla-4(12),8(13)dien-5-ol16361.87-0.87
50Hinesol1638-4.356.86
51β-Eudesmol1646-1.582.33
52α-Eudesmol16521.95--
53α-Cadinol1653--1.41
541(5),3-Aromadenedriene1660--0.51
5514- hydroxy-9-epi-β- Caryophyllene16640.970.27-
No.CompoundsaRIbHD(%)SD(%)SFME(%)
56Khusinol1674-0.31-
57(z)-Nerolidol acetate16750.79--
581,6-dimethyl-4-(1-methylethyl) Naphtalene1685-0.300.98
59Germacrone16930.86--
602-Pentadecanone1696--0.61
61(E)- Neroliol acetate17120.55--
62(E,E)- Farnesol1722--0.38
63Tetradecanoic acid17710.610.501.18
64Octadecane1800t0.21-
65Hexadecanal1806--0.64
666,10,14-trimethyl-2-Pentadecane18720.800.440.88
67Nonadecane19000.670.18-
68Hexadecanoic acid19737.302.766.55
69Eicosane20000.63--
70Henicosane2100-0.340.53
Monoterpene hydrocarbons0.7010.816.77
Oxygenated monoterpenes54.4457.8849.20
Sesquiterpene hydrocarbons-0.271.29
Oxygenated sesquiterpenes27.2713.6223.18
Other compounds14.468.7414.66
Total96.8791.3295.1

Compounds listed in order of elution from HP-5 MS column;

Retention indices to C8 - C24 n-alkanes on HP-5 MS column;

According to these results, the composition of the three oils show significant similarity for the concentration of the main components. All three oils were rich in regard tomonoterpenes (55.14%, 68.69%and 55.97%, respectively), while the sesquiterpenes fraction was (27.27%, 13.89% and 24.47%, respectively). The nonterpenoid fraction was relatively small, representing 14.46%, 8.74%, and 14.66%, respectively.

In our pervious investigations on Tanacetum genus we have identified essential oil compositions of T. balsamitha, T. polycephalum,T.khorassanicum,T. paradoxum,T. tabrisianum, T. elburensisand T.persicum(24-28). The dominant compound in T. balsamitha was carvone (68.0%) (24). Camphor (18.2% and 13.9%) and 1,8- cineole (17.0% and 18.6%) were found to be the major components of the oil of T. polycephalum and T. lingulatum, respectively (25, 29).

The major constituents of the aerial partsof T. khorassanicum were (E)-myroxide (19.8%), camphor (16.4%), isopulegon (13.4%) and 1,8-cineole (11.4%) (26).

Water distilled oil obtained from the aerial parts of T. paradoxum and T.tabrisianum have been the subject of our previous studies. The major components were camphor (23.8%), lavandulyl acetate (19.1%), lavandulol (15.9%) and 1,8- cineole (13.2%) in the former oil, caryophyllene oxide (12.0%)and spathulenol (10.3%) in the latter (27). Water- distilled oils from the aerial parts of T. elburensis and T. persicumgrowing wild in Iran were investigated .

The main constituents of the oil of T. elburensis were menthylisovalerate (20.0%) and 1,8 cineole (16.6%). The oil of T. persicum was characterized by higher amounts of borneol (24.3%), menthyl acetate (17.3%), isobornyl 2- methyl butyrate (16.0%) and artedouglasia oxide D (14.3%) (28).

The oils obtained by hydrodistillation of the leaves and flowers of T.dumosum growing wild in Iran were investigated. The main constituents of the leaves oil were borneol (27.9%), bornylacetate (18. 4%) and 1,8- cineole (17.5%), while the main components of the flower oil were isobornyl-2-methylbutanoate(41.1%) andtrans-linalyl oxideacetate (11.9%) (30).

Aerial parts of T. sonbolii, endemic in Iran,contained α- cadinol (35.3%),globulol (20.1%) and 1,8-cineole (8.6%) as major constituents (31).

Camphor (30.2%), (z)-chrysanthenylacetate (26.5%) and α- farnesene (11.1%) were found to be the major components from the root oil of T.partheniumfrom Iran. (32).

The dominant compound in the flower and stem oils ofT. chiliophyllum, from Turkey, was camphor (17.3% and 10.4%) respectively,while root oil of the plant was characterized with hexadecanoic acid (37.5 %) (37).

The dominant compound in the oil of T. nitens and T. argenteum from Turkey was 1,8 cineole (27.57%) and α- pinene (27.86%) respectively (38).

Baser et al. reported the oils from the flowers of T. zahlbruckneri and flowers and stems of T. tabrisianum from Turkey.

The flower oil of T. zahlbruckneri was characterised by germacreneD (29.7%) andspathulenol(12.0%).1,8-Cineole (17.6% and 22.5%) and hexadecnoic acid (10.3% and 8.0%) were the major constituents of the flower and stem oil of T. tabrisianum respectively (39).

The flower, stem and leaf oils of T. densum fromTurkey,were characterized with camphor (30.9%,25.7% and 27.7%, respectively) (40).

Lavandulol (21.5%) and 1,8- cineole (15.2 %) were identified as major components in the oil of T. gracile from LadakhHimalaya (India) (41).

Results of the antibacterial activity of the essential oil of Tanacetumtenuisectumare shown in Table 2.

Table 2

Antibacterial activity of essential oil of Tanacetumtenuisectum

MicroorganismsInhibition zone(mm)MICaMBCb
Staphyloccusaureus(ATCC 5923)116251250
Escherichia coil(ATCC 25922)136251250
Klebsiellapneumoniae(ATCC 27736)166251250
Entobacteraerogenes(ATCC 49469)16 625625
Bacillus subtilis(ATCC 9372)13625625
Bacillus creus(ATCC 6633)1612502500

Minimum inhibitory concentration;

Minimum bactericidal concentration .

Anti bacterial activity was determined against six bacteria.

The oil has shown maximum zone of inhibition against Klebsiellapneumoniae, Entobaceraerogenes and Bacillus creus. Staphylococcus aureus, Escherichia coli, Klebsiellapneumoniae, Entobacteraerogenes and Bacillus subtilis were the most sensitive bacteria to the essential oil (having MIC value 625 µg/mL).

Entobacteraerogenes and Bacillus subtilis have a minimum bactericidal concentration (MBC value 625 µg/mL). 1,8- Cineole and camphor are well- known chemicals having antibacterial potentials (42, 43).

The antibacterial effects of borneol were also reported (44). As aresult of these findings, antibacterial activity of T. tenuisectum oil could be attributed to 1,8 cineol, camphor and borneol. The present study confirms that there is a positive correlation between the chemical content of the oils and their antibacterial activities.

Acknowledgements

References

  • 1.

    Rechinger KH. Tanacetum. In: Rechinger KH, Hedge IC, editors. Flora Iranica. No.158, Compositae. Graz: Akadmische Druck and Verlagsanstalt; 1986. 214 p.

  • 2.

    Mozaffarian V. Dictionary of Iranian Plant Names. Tehran: Farhang Moaser; 1996. p. 534-537.

  • 3.

    Rohloff J, Mordal R, Dragland S. Chemotypical variation of tansy (Tanacetum vulgare L) from 40 different locations in Norway. J. Agric. Food Chem. 2004;52:1742-1748. [PubMed ID: 15030239].

  • 4.

    Ernst E, Pittler MH. The efficacy and safety of feverfew (Tanacetum parthenium L): an update of a systematic review. Public Health Nutr. 2000;3:509-514. [PubMed ID: 11276299].

  • 5.

    Sur R, Martin K, Liebel F, Lyte P, Shapiro S, Southall M. Anti-inflammatory activity of parthenolide – depleted feverfew (Tanacetum parthenium). Inflammopharmacology. 2009;17:42-49. [PubMed ID: 19112586].

  • 6.

    Park SJ, Shin HJ, Yun HS. Parthenolide inhibits TRIF- dependent signaling pathway of Toll- like receptors in RAW2647 macrophages. Mol. Cells. 2011;31:261-265. [PubMed ID: 21347702].

  • 7.

    Brown AMG, Edwards CM, Davey MR, Power JB, Lowe KC. Effects of extracts of Tanacetum species on human polymorphonuclearleucocyte activity in-vitro. Phytother. Res. 1997;11:479-484.

  • 8.

    Ghantous A, Sinjab A, Herceg Z, Darwiche N. Parthenolide; from plant shoots to cancer roots. Drug Discov. Today. 2013;18:894-905. [PubMed ID: 23688583].

  • 9.

    Habibi Z, Yousefi M, Shahriari F, Khalafi J, Ashabi MA. Chemical composition of the essential oil of Tanacetum turcomanicum and T canescens from Iran. Chem. Nat. Comp. 2009;43:93-95.

  • 10.

    Mohsenzadeh F, Chehregani A, Amiri H. Chemical composition, antibacterial activity and cytotoxicity of essential oils of Tanacetum parthenium in different developmental stages. Pharm. Biol. 2011;49:920-926. [PubMed ID: 21592001].

  • 11.

    Onozato T, Nakamura CV, Cortez DA, Dias Filho BP, Ueda- Nakamura T. Tanacetum vulgare: antiherpes virus activity of crude extract and the purified compound parthenolide. Phytother. Res. 2009;23:791-796. [PubMed ID: 19152371].

  • 12.

    Hethelyi E, Tetenyi P, Danos B, Koczka I. Phytochemical and antimicrobial studieson the essential oils of the Tanacetum vulgare clones by gas chromatography/ mass spectrometry. Herb Hung. 1991;30:82-90.

  • 13.

    Polatoğlu K, Karakoç OC, Gőkçe A, Gőren N. Insecticidal activity of Tanacetum chiliophyllum (Fisch & Mey) var monocephalum Grierson extracts and a new sesquiterpene lactone. Phytochem. Let. 2011;4:432-435.

  • 14.

    Tiuman TS, Ueda-Nakamura T, Garcia Cortez DA, Dias Filho BP, Morgado- Diaz JA, De Souza W, Nakamura CV. Antileishmanial activity of parthenolide asesquiterpene lactone Isolated from Tanacetum parthenium. Antimicrob. Agents Chemother. 2005;49:176-182. [PubMed ID: 15616293].

  • 15.

    lzumi E, Morello LG, Unda- NakamuraT, Yamada- Ogatta SF, Filho BPD, Cortez DAG, Ferreira ICP, Morgado-Diaz JA, Nakamura CV. Trypanosoma cruzi: antiprotozoal activity of parthenolide obtained from Tranacetum parthenium (L) Schultz Bip (Asteraceae Compositae) against epimastigote and amastigote forms. Exp. Parasitol. 2008;118:324-330. [PubMed ID: 17950283].

  • 16.

    Aljancic I, Vajs V, Bulatovic V, Menkovic N, Milosavljevic S. Parthenolide from the aerial parts of Tanacetum larvatum. Biochem. Syst. Ecol. 2001;29:655-657. [PubMed ID: 11336815].

  • 17.

    Mahmood U, Kaul VK, Singh B. Sesquiterpene and long chain ester from Tanacetum longifolium. Phytochem. 2002;61:913-917.

  • 18.

    Merfort I. Perspectives on sesquiterpene lactones in inflammation and cancer. Curr. Drug Targets. 2011;12:1560-1573. [PubMed ID: 21561425].

  • 19.

    Kreuger MR, Grootjans S, Biavatli MW, Vandenabeele P, D'Herde K. Sesquiterpene lactones as drugs with multiple targets in cancer treatment: focus on parthenolide. Anticancer Drugs. 2012;29:883-896.

  • 20.

    Rabito MF, Britta EA, Pelegrini BL, Scariot DB, Almeida MB, Ferreira ICP. In-vitro and in-vivo antileishmania activity of sesquiterpene lactone-rich dichloro methane fraction obtained from Tanacetum parthenium (L) Schultz-Bip. Exp. Parasitol. 2014;143:18-23. [PubMed ID: 24810433].

  • 21.

    Rosselli S, Bruno M, Raimondo FM, Spadaro V, Varol M, Koparal AT, Maggio A. Cytotoxic effect of eudesmanolides isolated from flowers of Tanacetum vulgare ssp siculum. Molecules. 2012;17:8186-8195. [PubMed ID: 22777187].

  • 22.

    Rustaiyan A, Zare K, Habibi Z, Hashemi M. Germacronolide from Tanacetum polycephalum. Phytochem. 1990;29:3022-3023.

  • 23.

    Rustaiyan A, Sedaghats S. Two new guaianolides from Tanacetum fruticulosum Ledeb. Proc. Acta Hort. 2005;677:65-69.

  • 24.

    Monfared A, Davarani S, Rustaiyan A, Masoudi S. Composition of the essential oil of Tanacetum balsmitha L ssp balsamitades (Schultz Bip) Grierson from Iran. J. Essent .Oil Res. 2002;14:1-2.

  • 25.

    Rustaiyan A, Mojab F, Salsali M, Masoudi S, Yari M. Composition of the essential oil of Tanacetum polycephalum Schultz. Bip. J. Essent. Oil Res. 1999;11:497-498.

  • 26.

    Majed Jabari T, Vatanpoor H, Rustaiyan A, Masoudi S, Monfared A. Composition of the essential oil of Tanacetum Khorassanicum (Krasch) Parsa a new species from Iran. J. Essent. Oil Res. 2002;14:380-381.

  • 27.

    Habibi Z, Biniyaz T, Ghodrati T, Masoudi S, Rustaiyan A. Volatile constituents of Tanacetum paradoxum Bornm and Tanacetum tabrisianum (Boiss) Sosn et Takht from Iran. J. Essent. Oil Res. 2007;19:11-13.

  • 28.

    Habibi Z, Hejazi Y, Alipour S, Masoudi S, Rustaiyan A. Essential oils of Tanacetum elburensis Mozaff and Tanacetum persicum (Boiss) Mozaff from Iran. J. Essend. Oil Res. 2007;19:310-312.

  • 29.

    Afsharypour S, Mosaffa Jahromy M. Constituents of the essential oil of Tanacetum lingulatum (Boiss) Bornm. J. Essent .Oil Res. 2003;15:74-76.

  • 30.

    Ghanbarian GA, Naseri M, Hatami A, Jafari E. Comparative essential oil composition of aerial parts of Tanacetum dumosum Boiss from southern Zagros, Iran. Nat. Prod. Res. 2015;29:197-200. [PubMed ID: 25370611].

  • 31.

    Firozy M, Talebpour Z, Sonboli A. Essential oil composition and antioxidant activities of the various extracts of Tanacetum sonboli Mozaff (Asteraceae) from Iran. Nat. Prod. Res. 2012;26:2204-2207. [PubMed ID: 22115413].

  • 32.

    Mojab F, Tabatabai SA, Naghdi-Badi HA, Nickavar B, Ghadyani F. Essential oil of the root of Tanacetum parthenium (L) Schulz Bip (Asteraceae) from Iran. Iran. J. Pharm. Res. 2007;6:291-293.

  • 33.

    Van den Dool H, Kratz PD. Generalization of the retention index system including linear temperature programmed gas- liquid partition chromatography. J. Chromatogr. 1963;11:463-471. [PubMed ID: 14062605].

  • 34.

    Massada Y. Analyses of Essential Oil by Gas Chromatography Mass Spectrometry. New York: Wiley; 1976.

  • 35.

    Adams RP. Identification of Essential Oil Components by Gas Chromatography/Quadrupole Mass Spectroscopy. Allured Publishing Corporation: Carol stream; 2001.

  • 36.

    Baron EJ, Finegold SM. Diagnostic Microbiology-Part 2 Method for Testing Antimicrobial Effectiveness. Springer: Berlin; 1991. p. 172-194.

  • 37.

    Polatoğu K, Demirci F, Demirci B, Gören N, Başer KHC. Essential oil composition and antimicrobial activities of Tanacetum chiliophyllum (Fisch & Mey) Schultz Bip var monocephalum Grierson from Turkey. Rec. Nat. Prod. 2012;6:184-188.

  • 38.

    Bagci E, Kocak A. Essentail oil composition of two endemic Tanacetum (T nitens (Boiss & Neo) Grierson and T argenteum (Lam) Willd subsp Argenteum (Asteraceae) taxa growing wild in Turkey. Indust. Crops Prod. 2010;31:542-545.

  • 39.

    Polatoğlu K, Demirci B, Demirci F, Gören N, Başer KHC. Essential oil composition of endemic Tanacetum zahlbruckneri (Nab) and Tanacetum tabrisianum (Boiss) Sosn and Takht from Turkey. Nat. Prod. Res. 2011;25:576-584. [PubMed ID: 21409718].

  • 40.

    Polatoğlu K, DemirciB, Demirci F, Gören N, Başer KHC. The essential oil composition of Tanacetum densum (Labill) Heywood ssp eginense Heywood from Turkey. Rec . Nat. Prod. 2012;6:402-406.

  • 41.

    Kitchlu S, Bakshi SK, Kaul MK, Bhan MK, Thapa RK, Agarwal SG. Tanacetum gracile Hook F & T a new source of lavandulol from Ladakh Himalaya (India). Flav. Fragr. J. 2006;21:690-692.

  • 42.

    Pattnaik S, Subramanyam VR, Bapaji M, Kole CR. Antibacterial and antifungal activity of aromatic constituents of essential oils. Microbios. 1997;89:39-46. [PubMed ID: 9218354].

  • 43.

    Tzakou O, Pitarokii D, Chinou IB, Harvala C. Composition and antimicrobial activity of the essential oil of Salvia ringens. Planta Med. 2001;67:81-83. [PubMed ID: 11270730].

  • 44.

    Vardar-Unlu G, Candan F, Sokmen A, Daferera D, Polissiou M, Sokmen M, Donmez E, Tepe B. Antimicrobial and antioxidant activity of the essential oil and methanol extracts of Thymus pectinatus Fischet Mey var pectinatus (Lamiaceae). J. Agri. Food Chem. 2003;51:63-67.