First Report on Screening of the Profiles of the Essential Oils and Volatiles from the Aerial Parts of Marrubium persicum Using Classical and Advanced Methods Prior to Gas Chromatographic Mass Spectrometric Determination

Author

Department of Chemistry, Shahrood Branch, Islamic Azad University, Shahrood, Iran

Abstract

The Lamiaceae family consists of a broad spectrum of medicinal plants involving Marrubium L. genus. Regarding the diverse pharmaceutical uses of the plants belonging to this genus, they can be considered as proper alternatives for chemical drugs having harmful effects. The present work aims to identify and characterize chemical compositions of the essential oils and volatiles from the aerial partsof Marrubium persicum C. A. Mey. as an herbal plant in Iran using classical hydrodistillation. To establish a comprehensive comparison between the traditional techniques and advanced ones, microwave-based extraction techniques namely MAHD as well as SFME have also been utilized. In another part of this project, the profiles related to the volatile fractions from the aerial parts of Marrubium persicum C. A. Mey.have been assessed and compared with the other categories. The main components in the hydrodistillation (HD) method were α-pinene (21.5%), spathulenol (19.5%), α-thujene (17.4%), while the headspace solid-phase microextraction(HS-SPME) profile mainly consisted of β-caryophyllene (14%), eugenol (11.2%) and methyl eugenol (10.2%). On the other hand, using the SFME approach spathulenol (25.4%), α-pinene (17.4%) and germacrene D (9.5%) were found as the most abundant constituents. Moreover, in the MAHD profile caryophyllene oxide (13.1%), δ-elemene (12.4%), camphene (8.5%) were respectively the predominant natural compounds. According to gas chromatographic-mass spectrometric determinations, a total of 40 compounds were recognized in the corresponding profiles totally covering 94.6-99.7% of the whole chemical compositions. Sesquiterpene hydrocarbons were recognized as the most frequent groups of natural compounds in the profiles of the advanced approaches, whereas in the traditional one monoterpene hydrocarbons were found to be the dominant constituting group.

Keywords


1. Mozaffarian V. A Dictionary of Iranian Plant Names, Farhang Moaser Press, Iran, 1996.

2. Zargari A. Medicinal Plants, University of Tehran Press, Tehran (In Persian). 1989.

3. Karioti A, Skaltsa H, Heilmann J, Sticher O. Acylated flavonoid and phenylethanoid glycosides from Marrubium velutinum. Phytochemistry.2003;64:655-660.

4. Karioti A, Heilmann J, Skaltsa H. Secondary metabolites from Marrubium velutinum, growing wild in Greece. Z Naturforsch B Chem Sci.2005;60:328-332.

5. Nicholas HJ. Isolation of marrubiin, a sterol, and a sesquiterpene from Marrubium vulgare. J Pharm Sci.1964;53:895-899.

6. Martin-Nizard F, Sahpaz S, Furman C, Fruchart JC, Duriez P, Bailleul F. Natural phenylpropanoids protect endothelial cells against oxidised LDL-induced cytotoxicity. Planta Med.2003;69:207-211.

7. Cechinel V. Advances and perspectives in the field of active natural products: Studies conducted at NIQFAR/UNIVALI. Quim Nova.2000;23:680-685.

8. Edziri H, Mastouri M, Aouni M, Verschaeve L. Polyphenols content, antioxidant and antiviral activities of leaf extracts of Marrubium deserti growing in Tunisia. South Afr J Bot.2012;80:104-109.

9. Edziri H, Mastouri M, Mahjoub MA, Ammar S, Mighri Z, Gutmann L, Aouni M. Antiviral activity of leaves extracts of Marrubium alysson L. J Med Plants Res.2011;5:360-363.

10. Hamedeyazdan S, Fathiazad F, Sharifi S, Nazemiyeh H. Antiproliferative activity of Marrubium persicum extract in the MCF-7 human breast cancer cell line. Asian Pac J Cancer Prev.2012;13:5843-5848.

11. de Oliveira AP, Santin JR, Lemos M, Klein Junior LC, Couto AG, da Silva Bittencourt CM, Cechinel Filho V, de Andrade SF. Gastroprotective activity of methanol extract and marrubiin obtained from leaves of Marrubium vulgare L. (Lamiaceae). J Pharm Pharmacol.2011;63:1230-1237.

12. Rigano D, Aviello G, Bruno M, Formisano C, Rosselli S, Capasso R, Senatore F, Izzo AA, Borrelli F. Antispasmodic effects and structure-activity relationships of labdane diterpenoids from Marrubium globosum ssp libanoticum. J Nat Prod.2009;72:1477-1481.

13. Schlemper V, Ribas A, Nicolau M, Cechinel V. Antispasmodic effects of hydroalcoholic extract of Marrubium vulgare on isolated tissues. Phytomedicine.1996;3:211-216.

14. de Souza MM, de Jesus RAP, Cechinel V, Schlemper V. Analgesic profile of hydroalcoholic extract obtained from Marrubium vulgare. Phytomedicine.1998;5:103-107.

15. Masoodi MH, Ahmed B, Zargar IM, Khan SA, Khan S, Singh P. Antibacterial activity of whole plant extract of Marrubium vulgare. Afr J Biotechnol.2008;7:86-87.

16. Rigano D, Formisano C, Basile A, Lavitola A, Senatore F, Rosselli S, Brtmo M. Antibacterial activity of flavonoids and phenylpropanoids from Marrubium globosum ssp libanoticum. Phytother Res.2007;21:395-397.

17. Ulukanli Z, Akkaya A. Antibacterial activities of Marrubium catariifolium and Phlomis pungens Var. Hirta grown wild in Eastern Anatolia, Turkey. Int J Agric Biol.2011;13:105-109.

18. Zarai Z, Kadri A, Ben Chobba I, Ben Mansour R, Bekir A, Mejdoub H, Gharsallah N. The in-vitro evaluation of antibacterial, antifungal and cytotoxic properties of Marrubium vulgare L. essential oil grown in Tunisia. Lipids Health Dis.2011;10.

19. Alberto Ortega-Ramirez L, Rodriguez-Garcia I, Manuel Leyva J, Reynaldo Cruz-Valenzuela M, Adriana Silva-Espinoza B, Gonzalez-Aguilar GA, Siddiqui MW, Fernando Ayala-Zavala J. Potential of medicinal plants as antimicrobial and antioxidant agents in food industry: A hypothesis. J Food Sci.2014;79:R129-R137.

20. Amessis-Ouchemoukh N, Madani K, Fale PLV, Luisa Serralheiro M, Araujo MEM. Antioxidant capacity and phenolic contents of some Mediterranean medicinal plants and their potential role in the inhibition of cyclooxygenase-1 and acetylcholinesterase activities. Ind Crop Prod.2014;53:6-15.

21. Boulila A, Sanaa A, Ben Salem I, Rokbeni N, M'Rabet Y, Hosni K, Fernandez X. Antioxidant properties and phenolic variation in wild populations of Marrubium vulgare L. (Lamiaceae). Ind Crop Prod.2015;76:616-622.

22. Jovin E, Toth A, Beara I, Balog K, Orcic D, Mimica-Dukic N. Chemical composition and antioxidant capacity of Marrubium peregrinum L. from Vojvodina (Serbia). Planta Med.2008;74:1046-1047.

23. Kadri A, Zarai Z, Ben Chobba I, Bekir A, Gharsallah N, Damak M, Gdoura R. Chemical composition and antioxidant activity of Marrubium vulgare L. essential oil from Tunisia. Afr J Biotechnol.2011;10:3908-3914.

24. Matkowski A, Piotrowska M. Antioxidant and free radical scavenging activities of some medicinal plants from the Lamiaceae. Fitoterapia.2006;77:346-353.

25. Meyre-Silva C, Cechinel-Filho V. A review of the chemical and pharmacological aspects of the genus Marrubium. Curr Pharm Des.2010;16:3503-3518.

26. Pukalskas A, Venskutonis PR, Salido S, de Waard P, van Beek TA. Isolation, identification and activity of natural antioxidants from horehound (Marrubium vulgare L.) cultivated in Lithuania. Food Chem.2012;130:695-701.

27. VanderJagt TJ, Ghattas R, VanderJagt DJ, Crossey M, Glew RH. Comparison of the total antioxidant content of 30 widely used medicinal plants of New Mexico. Life Sci.2002;70:1035-1040.

28. Zaabat N, Hay A-E, Michalet S, Darbour N, Bayet C, Skandrani I, Chekir-Ghedira L, Akkal S, Dijoux-Franca M-G. Antioxidant and antigenotoxic properties of compounds isolated from Marrubium deserti de Noe. Food Chem Toxicol.2011;49:3328-3335.

29. Argyropoulou A, Samara P, Tsitsilonis O, Skaltsa H. Polar constituents of Marrubium thessalum Boiss. & Heldr. (Lamiaceae) and their cytotoxic/cytostatic activity. Phytother Res.2012;26:1800-1806.

30. Ahmed B, Masoodi MH, Siddique AH, Khan S. A new monoterpene acid from Marrubium vulgare with potential antihepatotoxic activity. Nat Prod Res.2010;24:1671-1680.

31. Essawy SS, Abo-elmatty DM, Ghazy NM, Badr JM, Sterner O. Antioxidant and anti-inflammatory effects of Marrubium alysson extracts in high cholesterol-fed rabbits. Saudi Pharm J.2014;22:472-482.

32. Marrelli M, Conforti F, Rigano D, Formisano C, Bruno M, Senatore F, Menichini F. Cytotoxic properties of Marrubium globosum ssp libanoticum and its bioactive components. Nat Prod Commun.2013;8:567-569.

33. Shaheen F, Rasool S, Shah ZA, Soomro S, Jabeen A, Mesaik MA, Choudhary MI. Chemical constituents of Marrubium vulgare as potential inhibitors of nitric oxide and respiratory burst. Nat Prod Commun.2014;9:903-906.

34. Kirmizibekmez H, Atay I, Kaiser M, Yesilada E, Tasdemir D. In vitro antiprotozoal activity of extracts of five Turkish Lamiaceae species. Nat Prod Commun.2011;6:1697-1700.

35. Kirmizibekmez H, Atay I, Kaiser M, Yesilada E, Tasdemir D. In vitro antiprotozoal activity of organic and aqueous extracts of several Turkish Lamiaceae species. Planta Med.2011;77:1426-1426.

36. Daoudi A, Aarab L, Abdel-Sattar E. Screening of immunomodulatory activity of total and protein extracts of some Moroccan medicinal plants. Toxicol Ind Health.2013;29:245-253.

37. Michelis F, Tiligada E, Skaltsa H, Lazari D, Skaltsounis AL, Delitheos A. Effects of the flavonoid pilloin isolated from Marrubium cylleneum on mitogen-induced lymphocyte transformation. Pharm Biol.2002;40:245-248.

38. Al-Bakri AG, Afifi FU. Evaluation of antimicrobial activity of selected plant extracts by rapid XTT colorimetry and bacterial enumeration. J Microbiol Methods.2007;68:19-25.

39. Khalil A, Dababneh BF, Al-Gabbiesh AH. Antimicrobial activity against pathogenic microorganisms by extracts from herbal Jordanian plants. J Food Agric Environ.2009;7:103-106.

40. Masoodi M, Ali Z, Liang S, Yin H, Wang W, Khan IA. Labdane diterpenoids from Marrubium vulgare. Phytochem Lett.2015;13:275-279.

41. Petrovic S, Pavlovic M, Maksimovic Z, Milenkovic M, Couladis M, Tzakou O, Niketic M. Composition and antimicrobial activity of Marrubium incanum Desr. (Lamiaceae) essential oil. Nat Prod Commun.2009;4:431-434.

42. Robles-Zepeda RE, Velazquez-Contreras CA, Garibay-Escobar A, Galvez-Ruiz JC, Ruiz-Bustos E. Antimicrobial activity of northwestern mexican plants against Helicobacter pylori. J  Med Food.2011;14:1280-1283.

43. Karioti A, Skopeliti M, Heilmann J, Tsitsilonis O, Skaltsa H. Novel cytotoxic labdane diterpenes from Marrubium cylleneum. Planta Med.2006;72:1025-1025.

44. Karioti A, Skopeliti M, Tsitsilonis O, Heilmann J, Skaltsa H. Cytotoxicity and immunomodulating characteristics of labdane diterpenes from Marrubium cylleneum and Marrubium velutinum. Phytochemistry.2007;68:1587-1594.

45. Mohammadhosseini M. Chemical composition of the volatile fractions from flowers, leaves and stems of Salvia mirzayanii by HS-SPME-GC-MS. J Essent Oil-Bear Plants.2015;18:464-476.

46. Mohammadhosseini M. Chemical composition of the essential oils and volatile fractions from flowers, stems and roots of Salvia multicaulis Vahl. by using MAHD, SFME and HS-SPME methods. J Essent Oil-Bear Plants.2015;18:1360-1371.

47. Mohammadhosseini M, Akbarzadeh A, Hashemi-Moghaddam H, Shahnama M, Fahimi B, Azami S. Gas chromatographic-mass spectrometric analysis of volatiles obtained by HS-SPME-GC-MS technique from aerial parts of Ziziphora capitata L., and evaluation for biological activity. Oriental J Chem.2016;32:1439-1451.

48. Nekoei M, Mohammadhosseini M. Application of HS-SPME, SDME and cold-press coupled to GC/MS to analysis the essential oils of Citrus sinensis CV. Thomson Navel and QSRR study for prediction of retention indices by stepwise and genetic algorithm-multiple linear regression approaches. Anal Chem Lett.2014;4:93-103.

49. Nekoei M, Mohammadhosseini M. Chemical compositions of the essential oils from the aerial parts of Achillea wilhelmsii using traditional hydrodistillation, microwave assisted hydrodistillation and solvent-free microwave extraction methods: Comparison with the volatile compounds obtained by headspace solid-phase microextraction. J Essent Oil-Bear Plants.2016;19:59-75.

50. Mohammadhosseini M. A Comprehensive Review on New Methods for Processing, Separation and Identification of the Essential Oils, Islamic Azad University of Shahrood Press, Shahrood, Iran, 2016.

51. Mohammadhosseini M. Screening of the profiles of the essential oils from the aerial parts of Nepeta racemosa using classical and microwave-based methods: Comparison with the volatiles using headspace solid-phase micro-extraction. J. Herbal Drugs.2016;7:97-108.

52. Mohammadhosseini M, Mahdavi B, Akhlaghi H. Characterization and chemical composition of the volatile oils from aerial parts of Eryngium bungei Bioss. (Apiaceae) by using traditional hydrodistillation, microwave assisted hydrodistillation and head space solid phase microextraction methods prior to GC and GC/MS analyses: A comparative approach. J Essent Oil-Bear Plants.2013;16:613-623.

53. Mohammadhosseini M, Mahdavi B, Shahnama M. Chemical composition of essential oils from aerial parts of Ferula gummosa (Apiaceae) in Jajarm Region, Iran using traditional hydrodistillation and solvent-free microwave extraction methods: A comparative approach. J Essent Oil-Bear Plants.2015;18:1321-1328.

54. Mohammadhosseini M, Nekoei M. Chemical compositions of the essential oils and volatile compounds from the aerial parts of Ferula ovina using hydrodistillation, MAHD, SFME and HS-SPME methods. J Essent Oil-Bear Plants.2014;17:747-757.

55. Nekoei M, Mohammadhosseini M. Chemical composition of essential oils of Salvia leriifolia by three different extraction methods prior to gas chromatographic-mass spectrometric determination: comparison of HD with SFME and HS-SPME. J Essent Oil-Bear Plants.2016;19:In press.

56. Adams RP. Identification of Essential Oil Components by Gas Chromatography/Mass Spectrometry, Allured Publishing Co., Carol Stream, IL., USA, 2007.

57. Akhlaghi H, Nekoei M, Mohammadhosseini M, Motavalizadehkakhky A. Chemical composition of the volatile oils from the flowers, stems and leaves of Prangos latiloba Korov. using the head space solid phase microextraction method prior to analysis by gas chromatography-mass spectrometry. J Essent Oil-Bear Plants.2012;15:328-335.

58. Mohammadhosseini M, Pazoki A, Akhlaghi H. Chemical composition of the essential oils from flowers, stems and roots of Salvia multicaulis Vahl. growing wild in Iran. Chem Nat Comp.2008;44 127-128.

59. Nekoei M, Mohammadhosseini M, Akhlaghi H. Chemical composition of the volatile oils from the aerial parts of Artemisia annua L. (Asteraceae) by using head space solid phase microextraction and hydrodistillation methods prior to gas chromatographic-mass spectrometric determination: A comparative investigation. J Essent Oil-Bear Plants.2012;15:926-933.

60. Mohammadhosseini M, Zamani HA, Akhlaghi H, Nekoei M. Hydrodistilled volatile oil constituents of the aerial parts of Prangos serpentinica (Rech.f., Aell. Esfand.) Hernnstadt and Heyn from Iran and quantitative structure-retention relationship simulation. J Essent Oil-Bear Plants.2011;14:559-573.

61. Akhlaghi H, Shafaghat A, Mohammadhosseini M. Chemical composition of the essential oil from leaves of Biebersteinia multifida DC. growing wild in Iran. J Essent Oil-Bear Plants.2009;12:365-368.

62. Hashemi-Moghaddam H, Mohammadhosseini M, Basiri M. Optimization of microwave assisted hydrodistillation on chemical compositions of the essential oils from the aerial parts of Thymus pubescens and comparison with conventional hydrodistllation. J Essent Oil-Bear Plants.2015;18:884-893.

63. Hashemi-Moghaddam H, Mohammadhosseini M, Salar M. Chemical composition of the essential oils from the hulls of Pistacia vera L. by using magnetic nanoparticle-assisted microwave (MW) distillation: Comparison with routine MW and conventional hydrodistillation. Anal Methods.2014;6:2572-2579.

64. Mohammadhosseini M. Hydrodistilled volatile oils of the flowers of Salvia leriifolia Bench. and Salvia multicaulis Vahl. as two growing wild plants in Iran. Asian J Chem.2012;24:1432-1434.

65. Mohammadhosseini M, Pazoki A, Zamani HA, Akhlaghi H. Chemical composition of the essential oil from aerial parts of Ajuga chamaecistus Ging. subsp Scopria in brackish regions of Iran. J Essent Oil-Bear Plants.2011;14:101-105.

66. Mohammadhosseini M, Pazoki A, Zamani HA, Akhlaghi H, Nekoei M. Chemical composition of the essential oil from aerial parts of Senicio gallicus Chaix growing wild in Iran. J Essent Oil-Bear Plants.2010;13:704-709.

67. Motavalizadehkakhky AR, Shafaghat A, Zamani HA, Akhlaghi H, Mohammadhosseini M, Mehrzad J, Ebrahimi Z. Compositions and the in vitro antimicrobial activities of the essential oils and extracts of two Achillea species from Iran. J Med Plants Res.2013;7:1280-1292.

68. Shahnama M, Azami S, Mohammadhosseini M. Characterization of the essential oil and evaluation of antibacterial activity of methanolic extract of Stachys lavandulifolia Vahl. Int J Curr Microbiol App Sci.2015;4:275-283.

69. Akhlaghi H, Asl MRS, Mohammadhosseini M. Composition of the essential oil of Hymnocrater platystegius in Iran. Chem Nat Comp.2009;45:448-449.

70. Akhlaghi H, Shafaghat A, Salimi F, Mohammadhosseini M. GC/MS analysis of the essential oil from wild Stachys pubescens Ten. leaves from Northwest Iran. Anal Chem Lett.2011;1:325-327.

71. Mohammadhosseini M, Akbarzadeh A, Shafaghat A, Hashemi-Moghaddam H, Mohammadi Nafchi A, Ashouri H. Chemical composition of the essential oils from flowers and leaves of Marsdenia erecta using microwave assisted hydrodistillation technique. J Essent Oil-Bear Plants.2016;19:863-874.

72. Rahimi M, Karimi E, Nekoei M, Mohammadhosseini M. Hydro-distilled volatile oil constituents from the aerial parts of Satureja mutica and QSRR simulation by multiple linear regression. J Essent Oil-Bear Plants.2016;19:307-320.

73. Mohammadhosseini M. Hydrodistilled volatile oil from stems of Eryngium creticum Lam. in the marginal brackish regions of Semnan province by using gas chromatography combined with mass spectrometry. Asian J Chem.2013;25:390-392.

74. Mohammadhosseini M, Beiranvand M. Chemical composition of the essential oil from the aerial parts of Satureja hortensis as a potent medical plant using traditional hydrodistillation. J Chem Health Risks.2013;3:49-60.

75. Mohammadhosseini M, Akbarzadeh A, Hashemi-Moghaddam H, Mohammadi Nafchi A, Mashayekhi HA, Aryanpour A. Chemical composition of the essential oils from the aerial parts of Artemisia sieberi by using conventional hydrodistillation and microwave assisted hydrodistillation: A comparative study. J Essent Oil-Bear Plants.2016;19:32-45.

76. Grassia A, Senatore F, Arnold NA, Bruno M, Piozzi F, Rigano D, Formisano C. Chemical composition and antimicrobial activity of the essential oils from aerial parts of two Marrubium sp (Lamiaceae) growing wild in Lebanon. Pol J Chem.2006;80:623-628.

77. Tajbakhsh M, Khalilzadeh MA, Rineh A, Balou J. Essential oils of Marrubium anisodon C. Koch and Marrubium propinquum Fisch et CA Mey., growing wild in Iran. J Essent Oil Res.2008;20:161-162.

78. Morteza-Semnani K, Saeedi M, Babanezhad E. The essential oil composition of Marrubium vulgare L. from Iran. J Essent Oil Res.2008;20:488-490.

79. Weel KGC, Venskutonis PR, Pukalskas A, Gruzdiene D, Linssen JPH. Antioxidant activity of horehound (Marrubium vulgare L.) grown in Lithuania. Fett-Lipid.1999;101:395-400.

80. Novaes AP, Rossi C, Poffo C, Pretti E, Oliveira AE, Schlemper V, Niero R, Cechinel V, Burger C. Preliminary evaluation of the hypoglycemic effect of some Brazilian medicinal plants. Therapie.2001;56:427-430.

81. Demirci B, Baser KHC, Kirimer N. Composition of the essential oil of Marrubium bourgaei ssp caricum P.H. Davis. J Essent Oil Res.2004;16:133-134.

82. Javidnia K, Miri R, Soltani M, Khosravi AR. Constituents of the essential oil of Marrubium astracanicum Jacq. from Iran. J Essent Oil Res.2007;19:559-561.

83. Kaurinovic B, Vlaisavljevic S, Popovic M, Vastag D, Djurendic-Brenesel M. Antioxidant Properties of Marrubium peregrinum L. (Lamiaceae) essential oil. Molecules.2010;15:5943-5955.

84. Kirimer Ne, Kurkcuoglu M, Akgul G, Baser KHC, Abdel-Megeed A. Composition of the essential oil of Marrubium anisodon C. Koch of Turkish origin. Rec Nat Prod.2015;9:234-236.

85. Laouer H, Yabrir B, Djeridane A, Yousfi M, Beldovini N, Lamamra M. Composition, antioxidant and antimicrobial activities of the essential oil of Marrubium deserti. Nat Prod Commun.2009;4:1133-1138.

86. Zawislak G. The chemical composition of the essential oil of Marrubium vulgare L. from Poland. Farmacia.2012;60:287-292.

87. Kurkcuoglu M, Baser KHC, Tosun A, Dogan E, Duman H. Essential oil composition of an endemic species of Turkey: Marrubium bourgaei Boiss. ssp bourgaei (Labiatae). J Essent Oil Res.2007;19:34-36.

88. Belhattab R, Larous L, Figueiredo AC, Santos PAG, Costa MM, Barroso JG, Pedro LG. Essential oil composition and glandular trichomes of Marrubium vulgare L. growing wild in Algeria. J Essent Oil Res.2006;18:369-373.

89. Salama MM, Taher EE, El-Bahy MM. Molluscicidal and mosquitocidal activities of the essential oils of Thymus capitatus Hoff. Et Link. and Marrubium vulgare L. Rev Inst Med Trop Sao Paulo.2012;54:281-286.

90. Nik ZB, Mirza M. Composition of the essential oil of Marrubium astracanicum Jacq. J Essent Oil Res.2003;15:342-343.

91. Hamdaoui B, Wannes WA, Marrakchi M, Ben Brahim N, Marzouk B. Essential oil composition of two Tunisian horehound species: Marrubium vulgare L. and Marrubium aschersonii Magnus. J Essent Oil-Bear Plants.2013;16:608-612.

92. Khanavi M, Ghasemian L, Motlagh EH, Hadjiakhoondi A, Shafiee A. Chemical composition of the essential oils of Marrubium parviflorum Fisch & C. A. Mey. and Marrubium vulgare L. from Iran. Flav Fragr J.2005;20:324-326.

93. Lazari DM, Skaltsa HD, Constantinidis T. Essential oils of Marrubium velutinum Sm. and Marrubium peregrinum L., growing wild in Greece. Flav Fragr J.1999;14:290-292.

94. Masoudi S, Aghajani Z, Rustaiyan A, Feizbakhsh A, Kakhky AM. Volatile constituents of Teucrium persicum Boiss., Thymus caucasicus Willd. ex Ronniger subsp grossheimii (Ronniger) Jalas. and Marrubium crassidens Boiss. three Labiatae herbs growing wild in Iran. J Essent Oil Res.2009;21:5-7.

95. Zawislak G. Chemical composition of essential oils of Marrubium vulgare L. and Marrubium incanum Desr. grown in Poland. Chemija.2012;23:136-140.

96. Argyropoulou C, Skaltsa H. Identification of essential oil components of Marrubium thessalum Boiss. & Heldr., growing wild in Greece. Nat Prod Res.2012;26:593-599.

97. Nik ZB, Mirza M, Shahmir F. Essential oil of Marrubium cuneatum Russell and its secretory elements. Flav Fragr J.2004;19:233-235.

98. Sarikurkcu C, Tepe B, Daferera D, Polissiou M, Harmandar M. Studies on the antioxidant activity of the essential oil and methanol extract of Marrubium globosum subsp globosum (Lamiaceae) by three different chemical assays. Biores Technol.2008;99:4239-4246.