Artemia salina Toxicity Assay: Standardization in a 24‐Well Microplate

ABSTRACT

Bioassays with Artemia sp. are widely used in aquatic, environmental, and human toxicology as an initial tool to investigate the toxicity of plant extracts, isolated compounds, drugs, and products of natural or synthetic origin. These assays allow for the estimation of the median lethal concentration (LC ₅₀ ) and guide subsequent studies in cellular or animal models. Traditionally, two methods are used as a reference for the tests; however, experimental adaptations are necessary to optimize resources and overcome limitations such as the larger sample requirement and/or the difficulty in handling the larvae. This study aimed to standardize the bioassay with Artemia salina from nauplii collection to execution in 24‐well microplates, in order to evaluate its applicability as an alternative to conventional methods. The assay was performed in three independent experimental blocks, with exposure to nine DMSO concentrations. The materials used for collecting, counting, and transferring the nauplii proved to be adequate, allowing for quick and safe handling with minimal damage to the organisms. The LC ₅₀ obtained was 7.17%, a value consistent with reports in the literature. Statistical analysis confirmed that concentration was the main determinant of mortality. Thus, the proposed standardization proved to be viable, generating statistically and reproducible results, in addition to reducing sample consumption and optimizing the experimental process.