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Licensed Unlicensed Requires Authentication Published by De Gruyter May 23, 2022

Antibacterial, antioxidant, and cytotoxic activities of Syzygium aromaticum (L.) Merr. & Perry essential oil with identification of its chemical constituents

  • Wael A. Al-Zereini ORCID logo EMAIL logo , Ibrahim N. Al-Trawneh , Mahmoud A. Al-Qudah , Heba M. TumAllah , Zead H. Abudayeh and Tahani Hijazin


Aromatic plants embrace volatile compounds with efficiency in treating different diseases. In Jordan, Syzygium aromaticum flower buds (clove) are extensively used as folk medicine without awareness of its bio-safe dosage. Herein, clove buds were hydrodistilled using the Clevenger apparatus, and the resulting essential oil (CEO) was analyzed using Gas Chromatography-Mass Spectrometry (GC-MS). The antibacterial activity was evaluated against tested bacterial strains by agar diffusion test and micro-broth dilution assay. The antioxidant capacity was assessed using DPPH radical scavenging assay, while the cytotoxic potency was unraveled by determination of its anti-proliferative activity against MDA-MB-231 breast adenocarcinoma and normal Vero cell lines. CEO yield was 5.7 ± 1.3% (w/w); encompassed 24 volatile ingredients with eugenol as the principal compound (73.41%). The CEO inhibited the growth of both Gram-positive and Gram-negative bacterial test strains, causing the formation of 13.7 ± 1.5–17.3 ± 0.6 mm and 11.7 ± 1.5–20.7 ± 1.2 mm inhibition zones, respectively with MIC 1.25–5 μL/mL. Moreover, it showed antioxidant activity with IC50 0.0016 ± 0.0001 μL/mL (1.6 ± 0.1 μg/mL, 2.98 ± 0.4 µg Trolox®/µg CEO). Intriguingly, the CEO was cytotoxic against both cancerous and noncancerous cell lines at IC50 of 0.25 ± 0.02 μL/mL and 0.18 ± 0.01 μL/mL, respectively. Herein results unveil the potential application of CEO as a pharmaceutical remedy with considering its bio-safe dosage.

Corresponding author: Wael A. Al-Zereini, Department of Biological Sciences, Faculty of Science, Mutah University, Al-Karak 61710, Jordan, E-mail

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: No funding was received for this study.

  3. Conflicts of interest: The authors declare that there are no conflicts of interest.


1. Nasr, FA, Noman, OM, Mothana, RA, Alqahtani, AS, Al-Mishari, AA. Cytotoxic, antimicrobial and antioxidant activities and phytochemical analysis of Artemisia judaica and A. sieberi in Saudi Arabia. Afr J Pharm Pharmacol 2020;14:278–84.10.5897/AJPP2020.5175Search in Google Scholar

2. Al-Zereini, WA, Al-Trawneh, IN, Al-Qudah, MA, TumAllah, HM, Al Rawashdeh, HA, Abudayeh, ZH. Essential oils from Elettaria cardamomum (L.) Maton grains and Cinnamomum verum J. Presl barks: chemical examination and bioactivity studies. J Pharm Pharmacog Res 2022;10:173–85.10.56499/jppres21.1162_10.1.173Search in Google Scholar

3. Iqbal, J, Abbasi, BA, Mahmood, T, Kanwal, S, Ali, B, Shah, SA, et al.. Plant-derived anticancer agents: a green anticancer approach. Asian Pac J Trop Biomed 2017;7:1129–50. in Google Scholar

4. Che, CT, Zhang, H. Plant natural products for human health. Int J Mol Sci 2019;20:830. in Google Scholar PubMed PubMed Central

5. Cortés-Rojas, DF, de Souza, CRF, Oliveira, WP. Clove (Syzygium aromaticum): a precious spice. Asian Pac J Trop Biomed 2014;4:90–6. in Google Scholar

6. Vicidomini, C, Roviello, V, Roviello, GN. Molecular basis of the therapeutical potential of clove (Syzygium aromaticum L.) and clues to its anti-COVID-19 utility. Molecules 2021;26:1880. in Google Scholar PubMed PubMed Central

7. Behbahani, BA, Noshad, M, Falah, F. Study of chemical structure, antimicrobial, cytotoxic and mechanism of action of Syzygium aromaticum essential oil on foodborne pathogens. Potr S J F Sci 2019;13:875–83. in Google Scholar

8. Selles, SMA, Kouidri, M, Belhamiti, BT, Ait Amrane, A. Chemical composition, in-vitro antibacterial and antioxidant activities of Syzygium aromaticum essential oil. J Food Meas Char 2020;14:2352–8. in Google Scholar

9. Singh, NA, Kumar, P, Kumar, N. Spices and herbs: potential antiviral preventives and immunity boosters during COVID‐19. Phytother Res 2021;35:2745–57. in Google Scholar PubMed PubMed Central

10. Almuhur, RA. Routine usage of medicinal plants in northern Jordan. Pakistan J Nutr 2019;18:554–62. in Google Scholar

11. Mahmoud, I, Alkofahi, A, Abdelaziz, A. Mutagenic and toxic activities of several spices and some Jordanian medicinal plants. Int J Pharmacogn 1992;30:81–5. in Google Scholar

12. Abu-Ghazaleh, BM. Antimicrobial activity of Thymus vulgaris extract, Syzygium aromaticum extract and Zingiber officinale extract on Cronobacter spp. as compared with common preservatives. Pharmacology Online 2019;2:1–11.Search in Google Scholar

13. Ereifej, KI, Feng, H, Rababah, TM, Tashtoush, SH, Al-U’datt, MH, Al-Rabadi, GJ, et al.. Microbiological status and nutritional composition of spices used in food preparation. Food Nutr Sci 2015;6:1134–40. in Google Scholar

14. Sung, H, Ferlay, J, Siegel, RL, Laversanne, M, Soerjomataram, I, Jemal, A, et al.. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. Ca - Cancer J Clin 2021;71:209–49. in Google Scholar PubMed

15. International Agency for Research on Cancer/World Health Organization. Globocan 2020. Available online: [Accessed 17 Feb 2021].Search in Google Scholar

16. Adams, RP. Identification of Essential Oil Components by Gas Chromatography/Mass Spectrometry. Carol Stream, USA: Allured Publishing Corporation; 2007.Search in Google Scholar

17. Al-Zereini, WA. Bioactive crude extracts from four bacterial isolates of marine sediments from Red Sea, Gulf of Aqaba, Jordan. Jordan. J Biol Sci 2014;7:133–7. in Google Scholar

18. CLSI. Performance standards for antimicrobial susceptibility testing. In Twenty-second informational supplement, M100-S22. Wayne, USA: CLSI; 2012.Search in Google Scholar

19. Al-Qaralleh, OS, Al-Zereini, WA, Al-Mustafa, AH. Antibacterial, antioxidant and neuroprotective activities of crude extract from the endophytic fungus Fusarium sp. isolate OQ-Fus-2-F from Euphorbia sp. plant. J Pharm Pharmacog Res 2021;9:755–65.10.56499/jppres21.1077_9.6.755Search in Google Scholar

20. Al-Zereini, WA. Ononis natrix and Salvia verbenaca: two Jordanian medicinal plants with cytotoxic and antibacterial activities. J Herbs, Spices, Med Plants 2017;23:18–25. in Google Scholar

21. Finney, DJ. Probit analysis: a statistical treatment of the sigmoid response curve. Cambridge, UK: Cambridge University Press; 1947. p. 8–47.Search in Google Scholar

22. Saeed, A, Shahwar, D. Evaluation of biological activities of the essential oil and major component of Syzygium aromaticum. J Anim Plant Sci 2015;25:1095–9.Search in Google Scholar

23. El Ghallab, Y, Al Jahid, A, Jamal Eddine, J, Ait Haj Said, A, Zarayby, L, Derfoufi, S. Syzygium aromaticum L.: phytochemical investigation and comparison of the scavenging activity of essential oil, extracts and eugenol. Adv Tradit Med 2020;20:153–8. in Google Scholar

24. Mejía-Argueta, EL, Santillán-Benítez, JG, Canales-Martinez, MM, Mendoza-Medellín, A. Antimicrobial activity of Syzygium aromaticum L. essential oil on extended-spectrum beta-lactamases-producing Escherichia coli. Bull Natl Res Cent 2020;44:1–7. in Google Scholar

25. Teles, AM, Silva-Silva, JV, Fernandes, JMP, Abreu-Silva, AL, Calabrese, KDS, Mendes Filho, NE, et al.. GC-MS characterization of antibacterial, antioxidant, and antitrypanosomal activity of Syzygium aromaticum essential oil and eugenol. Evid base Compl Alternative Med 2021;2021:1–12. in Google Scholar PubMed PubMed Central

26. Hossain, MA, Harbi, SRA, Weli, AM, Al-Riyami, Q, Al-Sabahi, JN. Comparison of chemical constituents and antimicrobial activities of three essential oils from three different brands’ clove samples collected from Gulf region. Asian Pac J Trop Dis 2014;4:262–8. in Google Scholar

27. Alma, MH, Ertas, M, Nitz, S, Kollmannsberger, H. Chemical composition and content of essential oil from the bud of cultivated Turkish clove (Syzygium aromaticum L.). Bioresources 2007;2:265–9. in Google Scholar

28. Chaieb, K, Hajlaoui, H, Zmantar, T, Kahla‐Nakbi, AB, Rouabhia, M, Mahdouani, K, et al.. The chemical composition and biological activity of clove essential oil, Eugenia caryophyllata (Syzigium aromaticum L. Myrtaceae): a short review. Phytother Res 2007;21:501–6. in Google Scholar PubMed

29. Nana, WL, Eke, P, Fokom, R, Bakanrga‐Via, I, Begoude, D, Tchana, T, et al.. Antimicrobial activity of Syzygium aromaticum and Zanthoxylum xanthoxyloides essential oils against Phytophthora megakarya. J Phytopathol 2015;163:632–41. in Google Scholar

30. Radünz, M, da Trindade, MLM, Camargo, TM, Radünz, AL, Borges, CD, Gandra, EA, et al.. Antimicrobial and antioxidant activity of unencapsulated and encapsulated clove (Syzygium aromaticum, L.) essential oil. Food Chem 2019;276:180–6. in Google Scholar PubMed

31. Fuentes, JAM, Fernandez, IM, Aleman, RS, Maldonado, SAS, Roger, LF, Funez, NH, et al.. Chemical characterization of the essential oil of Syzygium aromaticum and its antimicrobial activity against a probiotic Lactobacillus acidophilus. Eur Sci J 2020;16:33–42. in Google Scholar

32. Boughendjioua, H. Essential oil composition of Syzygium aromaticum (L.). IRJPMS 2018;11:26–8.Search in Google Scholar

33. Razafimamonjison, G, Jahiel, M, Duclos, T, Ramanoelina, P, Fawbush, F, Danthu, P. Bud, leaf and stem essential oil composition of Syzygium aromaticum from Madagascar, Indonesia and Zanzibar. Int J Basic Appl Sci 2014;3:224–33.Search in Google Scholar

34. Kaur, K, Kaushal, S. Phytochemistry and pharmacological aspects of Syzygium aromaticum: a review. J Pharmacogn Phytochem 2019;8:398–406.Search in Google Scholar

35. Hamad, A, Mahardika, MGP, Yuliani, I, Hartanti, D. Chemical constituents and antimicrobial activities of essential oils of Syzygium polyanthum and Syzygium aromaticum. Rasayan J Chem 2017;10:564–9.Search in Google Scholar

36. Moemenbellah‐Fard, MD, Abdollahi, A, Ghanbariasad, A, Osanloo, M. Antibacterial and leishmanicidal activities of Syzygium aromaticum essential oil versus its major ingredient, eugenol. Flavour Fragr J. 2020;35:534–40.10.1002/ffj.3595Search in Google Scholar

37. Cai, L, Wu, CD. Compounds from Syzygium aromaticum possessing growth inhibitory activity against oral pathogens. J Nat Prod 1996;59:987–90. in Google Scholar PubMed

38. Kamatou, GP, Vermaak, I, Viljoen, AM. Eugenol—from the remote Maluku Islands to the international market place: a review of a remarkable and versatile molecule. Molecules 2012;17:6953–81. in Google Scholar PubMed PubMed Central

39. Zhang, K. Chemical composition and antioxidant activities of the essential oil from the clove buds (Syzygium aromaticum) toward various oxidative stresses in vitro. Agric Food Sci Res 2015;2:19–24.Search in Google Scholar

40. Abozid, MM, El-Sayed, SM. Antioxidant and protective effect of clove extracts and clove essential oil on hydrogen peroxide treated rats. Int J Chem Tech Res 2013;5:1477–85.Search in Google Scholar

41. Abd Al-azem, D, Al-Derawi, KH, Al-Saadi, SAM. The protective effects of Syzygium aromaticum essential oil extract against methotrexate induced hepatic and renal toxicity in rats. J Pure Appl Microbiol 2019;13:505–16. in Google Scholar

42. Halder, S, Mehta, AK, Kar, R, Mustafa, M, Mediratta, PK, Sharma, KK. Clove oil reverses learning and memory deficits in scopolamine-treated mice. Planta Med 2011;77:830–4. in Google Scholar PubMed

43. Angelini, P, Tirillini, B, Akhtar, MS, Dimitriu, L, Bricchi, E, Bertuzzi, G, et al.. Essential oil with anticancer activity: an overview. In Anticancer plants: Natural products and biotechnological implements; Akhtar, M, Swamy, M, Eds. Singapore: Springer, 2018; pp. 207–31.10.1007/978-981-10-8064-7_9Search in Google Scholar

44. Abdullah, ML, Hafez, MM, Al-Hoshani, A, Al-Shabanah, O. Anti-metastatic and anti-proliferative activity of eugenol against triple negative and HER2 positive breast cancer cells. BMC Compl Alternative Med 2018;18:1–11. in Google Scholar PubMed PubMed Central

45. Batiha, GES, Alkazmi, LM, Wasef, LG, Beshbishy, AM, Nadwa, EH, Rashwan, EK. Syzygium aromaticum L. (Myrtaceae): traditional uses, bioactive chemical constituents, pharmacological and toxicological activities. Biomolecules 2020;10:202. in Google Scholar PubMed PubMed Central

46. Jeng, JH, Hahn, LJ, Lu, EJ, Wang, YJ, Kuo, MYP. Eugenol triggers different pathobiological effects on human oral mucosal fibroblasts. J Dent Res 1994;73:1050–5. in Google Scholar PubMed

47. Prashar, A, Locke, IC, Evans, CS. Cytotoxicity of clove (Syzygium aromaticum) oil and its major components to human skin cells. Cell Prolif 2006;39:241–8. in Google Scholar PubMed PubMed Central

48. Ranjitkar, S, Zhang, D, Sun, F, Salman, S, He, W, Venkitanarayanan, K, et al.. Cytotoxic effects on cancerous and non-cancerous cells of trans-cinnamaldehyde, carvacrol, and eugenol. Sci Rep 2021;11:1–10. in Google Scholar PubMed PubMed Central

Received: 2022-03-14
Accepted: 2022-04-30
Published Online: 2022-05-23
Published in Print: 2023-03-28

© 2022 Walter de Gruyter GmbH, Berlin/Boston

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