Toxicity of Activated-carbon Purified Centella Asiatica (L.) Urb. Methanolic Extract on Zebrafish (Danio Rerio) Embryos

Abstract

Background: The development of novel purification methods using activated carbon on Centella asiatica methanolic extract, necessitates rigorous toxicity testing, as this method remains untested for toxicity and requires foundational safety data for subsequent validation and application stages. Objective: The study aims to identify the toxicity of activated-carbon purified Centella asiatica methanolic extract (ACP-CME) on Zebrafish (Danio rerio) embryos and determine its LC50. Method: Toxicity testing was performed by exposing zebrafish embryos to concentrations of 250, 125, 62.5, 31.25, and 12.5 ppm for 96 hours. The control groups included a positive control treated with 4 mg/L 3,4-DCA, a solvent control with 0.2% DMSO, and negative controls in sterile distilled water. The result data was analyzed using Microsoft Excel to calculate the LC50 and determine the toxicity signs of the extract. Result: ACP-CME exhibited concentration-dependent toxicity, with 100% mortality at 250 ppm and 10% mortality at 12.5 ppm. At concentrations of 31.25 ppm and 125 ppm, 10% and 25% of embryos, respectively, displayed abnormalities, including lordosis, kyphosis, delayed tail-bud detachment, tail abnormalities, somite formation failure, hatching failure, and yolk sac edema. Conclusion: The LC50 of ACP-CME is 38.74 ppm, while the observed toxicity is characterized by mortality, lordosis, kyphosis, delayed tail-bud detachment, abnormalities of the tail, failure on somite formation, hatching failure, and yolk sac edema.