Skip to main navigation menu Skip to main content Skip to site footer

Research Articles

Vol. 13 No. 3 (2026)

Phytochemical analysis, in vitro antioxidant and antibacterial activities and embryotoxicity evaluation of the essential oil of Microtoena patchoulii (C.B.Clarke ex Hook.f.) C.Y.Wu & S.J.Hsuan and terpinolene in zebrafish (Danio rerio)

DOI
https://doi.org/10.14719/pst.12427
Submitted
26 October 2025
Published
09-07-2026 — Updated on 18-08-2026
Versions

Abstract

Patchouli oil has diverse applications in food preservation, medicine and cosmetics. The present study investigated the phytochemical composition, antioxidant and antibacterial activities and embryotoxic potential of the essential oil of Microtoena patchoulii (C.B.Clarke ex Hook.f.) C.Y.Wu & S.J.Hsuan (EOMP). Phytochemical profiling was performed using gas chromatography-mass spectrometry (GC-MS) and Fourier transform infrared spectroscopy (FTIR). Antioxidant activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assays. Antibacterial activity of EOMP and terpinolene was assessed by the agar well diffusion method against Escherichia coli, Bacillus subtilis, Salmonella typhi and Staphylococcus aureus. Minimum inhibitory concentrations (MICs) were determined using a 96-well microtiter plate assay with tetrazolium dye. Embryotoxicity was evaluated in zebrafish (Danio rerio). The GC-MS analysis identified terpinolene (10.15 %) as the major volatile constituent. The EOMP exhibited stronger ABTS radical scavenging activity (EC₅₀ = 6.995 µg/mL) than DPPH activity (EC₅₀ = 10.044 µg/mL). The antibacterial activity of EOMP was observed in the order: E. coli (24.6 ± 0.1 mm) > S. aureus (22.0 ± 0.1 mm) > S. typhi (20.4 ± 0.6 mm) > B. subtilis (18.6 ± 0.1 mm), with the lowest MIC recorded against E. coli (5.0 ± 0.0 µg/mL). Terpinolene showed comparatively lower antibacterial activity, with the lowest MIC observed against B. subtilis and S. aureus (2.0 ± 0.0 µg/mL). The median lethal concentration (LC₅₀) values of EOMP and terpinolene were 174.147 µg/mL and 201.409 µg/mL, respectively. Overall, EOMP demonstrated stronger antibacterial activity but higher embryotoxic potential than terpinolene.

References

  1. 1. Livermore DM. Discovery research: the scientific challenge of finding new antibiotics. J Antimicrob Chemother. 2011;66:1941–4. https://doi.org/10.1093/jac/dkr262
  2. 2. Bakkali F, Averbeck S, Averbeck D, Idaomar M. Biological effects of essential oils: a review. Food Chem Toxicol. 2008;46:446–75. https://doi.org/10.1016/j.fct.2007.09.106
  3. 3. Wang Q, Hong DY. Character analysis and taxonomic revision of the Microtoena insuavis complex (Lamiaceae). Bot J Linn Soc. 2011;165:315–27. https://doi.org/10.1111/j.1095-8339.2010.01111.x
  4. 4. Senpuku M, Nonaka K, Ito M, Honda G. Chemical composition of the essential oil of Microtoena patchoulii [(C.B. Clarke ex J.D. Hooker) C.Y. Wu et Hsuan]. J Essent Oil Res. 2007;19:336–7. https://doi.org/10.1080/10412905.2007.9699297
  5. 5. Khomdram SD, Singh PK. Harnessing the economic potential of an aromatic Lamiaceae plant Microtoena patchoulii (Clarke ex Hook.) Wu & Hsuan in Manipur, India. Recent Adv Nat Prod Res. 2012.
  6. 6. Hu G, Peng C, Xie X, Zhang S, Cao X. Availability, pharmaceutics, security, pharmacokinetics and and pharmacological activities of patchouli alcohol. Evid Based Complement Alternat Med. 2017;2017:4850612. https://doi.org/10.1155/2017/4850612
  7. 7. Lanzerstorfer P, Sandner G, Pitsch J, Mascher B, Aumiller T, Weghuber J. Acute, reproductive and and developmental toxicity of essential oils assessed with alternative in vitro and in vivo systems. Arch Toxicol. 2020;95:673–91. https://doi.org/10.1007/s00204-020-02945-6
  8. 8. Jayasinghe CD, Jayawardena UA. Toxicity assessment of herbal medicine using zebrafish embryos: a systematic review. Evid Based Complement Alternat Med. 2019;2019:7272808. https://doi.org/10.1155/2019/7272808
  9. 9. OECD. Guidelines for the testing of chemicals. Section 2: effects on biotic systems, test No. 236: fish embryo acute toxicity (FET) test. Paris: Organisation for Economic Co-operation and Development; 2013.
  10. 10. Brand-Williams W, Cuvelier ME, Berset C. Use of a free radical method to evaluate antioxidant activity. LWT Food Sci Technol. 1995;28:25–30. https://doi.org/10.1016/S0023-6438(95)80008-5
  11. 11. Dawidowicz AL, Olszowy M. Antioxidant properties of BHT estimated by ABTS assay in systems differing in pH or metal ion or water concentration. Eur Food Res Technol. 2011;232:837–42. https://doi.org/10.1007/s00217-011-1451-7
  12. 12. Miller NJ, Sampson J, Candeias LP, Bramley PM, Rice-Evans CA. Antioxidant activities of carotenes and xanthophylls. FEBS Lett. 1996;384:240–2. https://doi.org/10.1016/0014-5793(96)00323-7
  13. 13. Ashraf SA, Al-Shammari E, Hussain T, Tajuddin S, Panda BP. In vitro antimicrobial activity and identification of bioactive components using GC-MS of commercially available essential oils in Saudi Arabia. J Food Sci Technol. 2017;54:3948–58. https://doi.org/10.1007/s13197-017-2859-2
  14. 14. Perumal S, Pillai S, Cai LW, Mahmud R, Ramanathan S. Determination of minimum inhibitory concentration of Euphorbia hirta (L.) extracts by tetrazolium microplate assay. J Nat Prod. 2012;5:68–76.
  15. 15. Kimmel CB, Ballard WW, Kimmel SR, Ullmann B, Schilling TF. Stages of embryonic development of the zebrafish. Dev Dyn. 1995;203:253–310. https://doi.org/10.1002/aja.1002030302
  16. 16. Mrisho II, Musazade E, Chen H, Zhao H, Xing J, Li X, et al. Unlocking the therapeutic potential of patchouli leaves: a comprehensive review of phytochemical and pharmacological insights. Plants. 2025;14:1034. https://doi.org/10.3390/plants14071034
  17. 17. Li YQ, Kong DX, Wu H. Analysis and evaluation of essential oil components of cinnamon barks using GC-MS and FTIR spectroscopy. Ind Crops Prod. 2013;41:269–78. https://doi.org/10.1016/j.indcrop.2012.04.056
  18. 18. Hosseini SF, Zandi M, Rezaei M, Farahmandghavi F. Two-step method for encapsulation of oregano essential oil in chitosan nanoparticles: preparation, characterization and in vitro release study. Carbohydr Polym. 2013;95:50–6. https://doi.org/10.1016/j.carbpol.2013.02.031
  19. 19. Wen P, Zhu DH, Wu H, Zong MH, Jing YR, Han SY. Encapsulation of cinnamon essential oil in electrospun nanofibrous film for active food packaging. Food Control. 2016;59:366–76. https://doi.org/10.1016/j.foodcont.2015.06.005
  20. 20. Wei A, Shibamoto T. Antioxidant and lipoxygenase inhibitory activities and chemical compositions of selected essential oils. J Agric Food Chem. 2007;55:1737–42. https://doi.org/10.1021/jf101077s
  21. 21. Zuo GY, et al. Antibacterial and synergistic effects of pogostone from Pogostemon cablin (patchouli) against drug-resistant pathogenic bacteria. J Ethnopharmacol. 2012;140:214–20. https://doi.org/10.1016/j.jep.2012.01.027
  22. 22. Gushiken LFS, Beserra FP, Hussni MF, Gonzaga MT, Ribeiro VP, de Souza PF, et al. Beta-caryophyllene as an antioxidant, anti-inflammatory and re-epithelialization agent in a rat skin wound excision model. Oxid Med Cell Longev. 2022;2022:9004014. https://doi.org/10.1155/2022/9004014
  23. 23. da Silva II Jr, da Silva NPC, Marrs JA, Cadena PG. Essential oils produce developmental toxicity in zebrafish embryos and cause behavior changes in zebrafish larvae. Biomedicines. 2023;11:2821. https://doi.org/10.3390/biomedicines11102821
  24. 24. Sisman T, Ceylan Z. The embryotoxicity of alpha-pinene in zebrafish (Danio rerio Hamilton, 1822). Nat Life Sci Commun. 2023. https://doi.org/10.12982/NLSC.2023.020
  25. 25. Szaszkiewicz J, Leigh S, Hamilton TJ. Robust behavioural effects in response to acute, but not repeated, terpene administration in zebrafish (Danio rerio). Sci Rep. 2021;11:19214. https://doi.org/10.1038/s41598-021-98768-1

Downloads

Download data is not yet available.