Acute Toxicity Tests of Fermented Robusta Green Coffee Using Zebrafish Embryos (Danio rerio)

Background: Green coffee beans are coffee beans of coffee fruit that have not yet been roasted. The use of green coffee beans as a weight-loss agent has been widely used worldwide, but nowadays there is a new way to enjoy coffee by adding kombucha culture to it, or what is known as kombucha coffee. The development of this fermented product preparation still requires a more in-depth study, one of which is related to the toxicity of the kombucha green coffee preparation. Objective: This research was aimed to determine LC₅₀ values for robusta green coffee fermented with kombucha culture using zebrafish (Danio rerio) embryos using an in vivo method. Methods: This study observed the development of 20 zebrafish embryos administered one of five different concentrations of kombucha coffee preparation 24 hours up to 96 hours, with the experiment repeated three times. The percentage of embryo deaths was observed and analyzed using a probit model for LC₅₀ concentration and analyzed using IBM SPSS Ver 23. Results: An LC₅₀ for kombucha green coffee of 1294.29 ppm was obtained. Statistical tests on the concentration of kombucha coffee extracts were not significant differences with p-values > 0.05. Conclusion: The LC₅₀ of robusta green coffee extract using zebrafish embryos of 1294.29 ppm included in the safe category.