Developmental Toxicity of Methanol in Zebrafish Embryos for Establishing Safe Solvent Concentrations

Authors

  • Siti Nurhafizah Ramli Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, D.E, Malaysia.
  • Noor Azmi Shaharuddin Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, D.E, Malaysia.
  • Syahida Ahmad Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, D.E, Malaysia.
  • Siti Nor Akmar Abdullah Department of Agricultural Technology, Faculty of Agriculture, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
  • Azzreena Mohamad Azzeme Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, D.E, Malaysia.

DOI:

https://doi.org/10.54987/jobimb.v13i1.1087

Keywords:

Methanol toxicity, Zebrafish embryo assay, Developmental teratogenicity, Safe solvent concentration, Bioassay solvent control

Abstract

Methanol (MeOH) is widely used as a solvent for extracting bioactive compounds from plant materials, but its inherent toxicity may confound biological assays if not properly controlled. This study aimed to evaluate the developmental toxicity of MeOH in zebrafish embryos to determine the maximum concentration that can be safely used as a solvent without causing adverse developmental effects. Zebrafish embryos were exposed to a range of MeOH concentrations (0.39% to 24.98% v/v in embryo media). Survival rate, mortality, hatching rate, heartbeat rate, and teratogenic effects were assessed over 120 hours post-fertilization (hpf). Embryo survival decreased with increasing MeOH concentrations. The highest survival (91.7%) was observed at 0.39% MeOH, followed by 83.3% at 0.78%, and 50% at 1.56%. Concentrations ≥3.13% caused 100% mortality. The calculated LC50 was 1.58% MeOH. Hatching and heartbeat rates followed similar concentration-dependent trends. Teratogenic effects, including oedema, failure of tail detachment, and curved body, were observed at sub-lethal concentrations. Methanol exhibits significant developmental toxicity in zebrafish embryos at concentrations above 1.56%. This study establishes 1.56% MeOH as the maximum concentration tolerable without severe toxicity, providing a safe threshold for its use as a solvent in future bioassays involving plant extracts or other biological samples.

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Published

31.07.2025

How to Cite

Ramli, S. N., Shaharuddin, N. A., Ahmad, S. ., Abdullah, S. N. A., & Azzeme, A. M. (2025). Developmental Toxicity of Methanol in Zebrafish Embryos for Establishing Safe Solvent Concentrations. Journal of Biochemistry, Microbiology and Biotechnology, 13(1), 117–122. https://doi.org/10.54987/jobimb.v13i1.1087

Issue

Vol. 13 No. 1 (2025)

Section

Articles

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Copyright (c) 2025 Siti Nurhafizah Ramli, Noor Azmi Shaharuddin, Syahida Ahmad, Siti Nor Akmar Abdullah, Azzreena Mohamad Azzeme

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