| Summary: | Benthic invertebrates produce secondary metabolites that serve as defenses against consumers and promote their fitness. To explore the chemical defense in marine benthic echinoderms, the chemical constituents of <i>Ophiopholis mirabilis</i> were investigated. A steroidal monoglycoside, asterosaponin P1, was isolated from <i>O. mirabilis</i> for the first-time using column chromatography. The chemical structure was characterized by spectroscopic and spectrometric methods. The embryotoxicity and teratogenicity of the isolated compound were assessed using the zebrafish embryo assay, a powerful vertebrate animal model system to study mechanisms of toxicity. When applied at high concentrations, asterosaponin P1 causes a significant increase in embryo mortality. A moderate LC<sub>50</sub> of asterosaponin P1 appeared to be time- and concentration-dependent in its toxicity to zebrafish embryos. Teratogenicity in zebrafish embryos also included morphological defects, decreased hatchability, and a reduced heart rate. These findings revealed that steroidal saponin extracted from <i>O. mirabilis</i> exhibited acute toxic effects on zebrafish embryos, suggesting a potential chemical defense function in marine habitats.
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