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

Research Articles

Early Access

Phytochemical characterisation and biosafety profiling of Elsholtzia communis (Collett & Hemsl.) Diels essential oil using zebrafish embryos

DOI
https://doi.org/10.14719/pst.13082
Submitted
5 December 2025
Published
22-03-2026
Versions

Abstract

Elsholtzia communis (Collett & Hemsl.) Diels is traditionally used in herbal medicine, yet the phytochemical composition and developmental safety of its flower-derived essential oil remain insufficiently explored. This study aimed to characterise the essential oil of E. communis flowers (EOEC), evaluate its antioxidant and antibacterial activities and assess its developmental toxicity using zebrafish (Danio rerio) embryos. The EOEC was extracted by hydro-distillation and characterised using gas chromatography-mass spectrometry (GC-MS) and attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy. Antioxidant potential was evaluated through DPPH and ABTS radical scavenging assays, while antibacterial activity against selected Gram-positive and Gram-negative bacteria was determined using agar well diffusion and minimum inhibitory concentration (MIC) assays. Developmental toxicity was assessed in zebrafish embryos by monitoring mortality and morphological abnormalities to calculate LC₅₀ values. The GC-MS analysis revealed citral-A (geranial, 45.5 %) and citral-B (neral, 37.8 %) as the major constituents of EOEC, while FTIR spectra confirmed the presence of characteristic OH, C=O and CH functional groups. EOEC exhibited moderate antioxidant activity with EC₅₀ values of 45.12 µg/mL (DPPH) and 43.15 µg/mL (ABTS) and demonstrated significant antibacterial effects against both Gram-positive and Gram-negative strains. Zebrafish embryo assays showed clear dose-dependent lethality and developmental abnormalities, with LC₅₀ values of 123 µg/mL for EOEC and 145 µg/mL for citral. Notably, adverse developmental effects occurred at concentrations close to those producing biological activity. In conclusion, EOEC possesses promising antioxidant and antibacterial properties but exhibits concentration-dependent embryotoxicity, emphasising the need for careful dose optimisation and biosafety evaluation prior to therapeutic or nutraceutical applications.

References

  1. 1. Singh HB, Singh RS, Sandhu JS. Herbal medicine of Manipur: a colour encyclopaedia. New Delhi: Daya Publishing House; 2003.
  2. 2. Nath S, Tamuli KJ, Saikia S, Narzary B, Gogoi B, Bordoloi M, et al. Essential oil from the leaves of Elsholtzia communis (Collett & Hemsl.) Diels from North East India: chemical profiling, antimicrobial, cytotoxic and ACE inhibitory activities. Flavour Fragr J. 2021;36:626c36. https://doi.org/10.1002/ffj.3677
  3. 3. U.S. Food and Drug Administration. Substances generally recognized as safe (GRAS): citral. Silver Spring (MD): FDA; 2023.
  4. 4. Jayasinghe CD, Jayawardene 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
  5. 5. National Toxicology Program. Toxicology and carcinogenesis studies of citral (microencapsulated) in F344/N rats and B6C3F1 mice (feed studies). Natl Toxicol Program Tech Rep Ser. 2003;(505):1–268.
  6. 6. Hill AJ, Teraoka H, Heideman W, Peterson RE. Zebrafish as a model vertebrate for investigating chemical toxicity. Toxicol Sci. 2005;86(1):6–19. https://doi.org/10.1093/toxsci/kfi110
  7. 7. 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
  8. 8. Dawidowicz AL, Olszowy M. Antioxidant properties of BHT estimated by ABTS assay in systems differing in pH, metal ion or water concentration. Eur Food Res Technol. 2011;232:837–42. https://doi.org/10.1007/s00217-011-1451-7
  9. 9. Hudzicki J. Kirby–Bauer disk diffusion susceptibility test protocol. Washington (DC): American Society for Microbiology; 2009.
  10. 10. 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. J Food Sci Technol. 2017;54:3948–58. https://doi.org/10.1007/s13197-017-2859-2
  11. 11. Perumal S, Pillai S, Cai LW, Mahmud R, Ramanathan S. Determination of minimum inhibitory concentration of Euphorbia hirta extracts by tetrazolium microplate assay. J Nat Prod. 2012;5:68–76.
  12. 12. 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
  13. 13. Doimo L. Iso-citral and iso-geraniols in lemon-myrtle (Backhousia citriodora F. Muell.) essential oils. J Essent Oil Res. 2001;13(4):236–7. https://doi.org/10.1080/10412905.2001.9699680
  14. 14. Oliveira GLS, Machado KC, Silva APDSCL, Feitosa CM, Almeida FRC. Non-clinical toxicity of β-caryophyllene: absence of adverse effects in female Swiss mice. Regul Toxicol Pharmacol. 2018;92:338–46. https://doi.org/10.1016/j.yrtph.2017.12.013
  15. 15. Achimón F, Peschiutta ML, Brito VD, Ulla SB, Pizzolitto RP. Sulcatone as a plant-derived volatile organic compound for control of maize weevil and associated phytopathogenic fungi in stored maize. Plants. 2024;13(20):2893. https://doi.org/10.3390/plants13202893
  16. 16. Hartsel JA, Eades J, Hickory B, Makriyannis A. Cannabis sativa and hemp. In: Watson RR, Preedy VR, Zibadi S, editors. Nutraceuticals. Academic Press; 2016. p. 735–54. https://doi.org/10.1016/B978-0-12-802147-7.00053-X
  17. 17. Li YQ, Kong DX, Wu H. Analysis and evaluation of essential oil components of cinnamon bark 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. Encapsulation of oregano essential oil in chitosan nanoparticles: preparation, characterization and in vitro release. 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. De Luca E, Zaccaria GM, Hadhoud M, Rizzo G, Ponzini R, Morbiducci U, et al. ZebraBeat: a flexible platform for analysis of cardiac rate in zebrafish embryos. Sci Rep. 2014;4:4898. https://doi.org/10.1038/srep04898
  21. 21. Murugesan R, Vasuki K, Balasubramanian K, Ramadevi S. Developmental toxicity and behavioral safety of ethanolic extract of Pedalium murex on zebrafish embryos. ACBR. 2023;:1958. https://doi.org/10.37256/acbr.2120231958
  22. 22. Mangalagiri NP, Velagapudi K, Panditi SK, Jeevigunta NLL. Mechanism of action of essential oils and their major components. Res Rev J Bot. 2021;10(3):33–43.
  23. 23. Di Pasqua R, Betts G, Hoskins N, Edwards M, Ercolini D, Mauriello G. Membrane toxicity of antimicrobial compounds from essential oils. J Agric Food Chem. 2007;55(12):4863–70. https://doi.org/10.1021/jf0636465
  24. 24. Boukhatem MN, Ferhat MA, Kameli A, Saidi F, Tchoketch Kebir H. Lemon grass (Cymbopogon citratus) essential oil as an anti-inflammatory and antifungal agent. Libyan J Med. 2014;9:25431. https://doi.org/10.3402/ljm.v9.25431
  25. 25. Zakaria ZZ, Benslimane FM, Nasrallah GK, Shurbaji S, Younes NN, Mraiche F, et al. Using zebrafish to investigate molecular mechanisms of drug-induced cardiotoxicity. Biomed Res Int. 2018;2018:1642684. https://doi.org/10.1155/2018/1642684
  26. 26. He JH, Gao JM, Huang CJ, Li CQ. Zebrafish models for assessing developmental and reproductive toxicity. Neurotoxicol Teratol. 2014;42:35–42. https://doi.org/10.1016/j.ntt.2014.01.006
  27. 27. Bertelli PR, Biegelmeyer R, Rico EP, Klein-Junior LC, Toson NSB, Minetto L, et al. Toxicological profile and acetylcholinesterase inhibitory potential of Palicourea deflexa. Comp Biochem Physiol C Toxicol Pharmacol. 2017;201:44–50. https://doi.org/10.1016/j.cbpc.2017.09.003
  28. 28. OECD. OECD guideline for the testing of chemicals: fish embryo toxicity (FET) test. Paris: OECD; 2006.
  29. 29. Chen L, Xu M, Gong Z, Zonyane S, Xu S, Makunga NP. Cardio- and developmental toxicity of Sutherlandia frutescens detected using zebrafish embryos. BMC Complement Altern Med. 2018;18:230. https://doi.org/10.1186/s12906-018-2303-9
  30. 30. Tolentino JJV. Embryotoxicity and teratogenicity of Derris elliptica leaf extract on zebrafish embryos. Int J Pure Appl Biosci. 2016;4:16–20. https://doi.org/10.18782/2320-7051.2293
  31. 31. Alafiatayo AA, Lai KS, Syahida A, Mahmood M, Shaharuddin NA. Phytochemical evaluation and embryotoxicity of Curcuma longa extract on zebrafish. Evid Based Complement Alternat Med. 2019;2019:3807207. https://doi.org/10.1155/2019/3807207
  32. 32. Ali MK, Saber SP, Taite DR, Emadi S, Irving R. Protective layer changes during zebrafish embryo development. J Toxicol Pharmacol. 2017;1:009.
  33. 33. Mektrirat R, Yano T, Okonogi S, Katip W, Pikulkaew S. Safety evaluation of volatile terpenoids from Zingiber cassumunar Roxb. using zebrafish embryos. Molecules. 2020;25:613. https://doi.org/10.3390/molecules25030613
  34. 34. da Silva Jr II, Correia da Silva NP, Marrs JA, Cadena PG. Essential oils induce developmental toxicity and behavioral changes in zebrafish. Biomedicines. 2023;11(10):2821. https://doi.org/10.3390/biomedicines11102821

Downloads

Download data is not yet available.