Micropollutants are compounds used to enhance human life standards that are present in the environment in ng/L to µg/L concentrations. The most common micropollutants present in the environment include pharmaceuticals, personal care products, pesticides and endocrine-disrupting compounds. The presence of many of these micropollutants has been reported in various aquatic matrices. Even though low concentrations of micropollutants are reported in the environment, their exposure result in significant toxicological concerns (short-term to long-term effects) for aquatic organisms and human health. This is because of their extended lifespan and multigenerational exposure to continuously produced and discharged micropollutants in the environment. Thus, it is pivotal to understand the toxic effects of micropollutants on the health of aquatic organisms and humans. In this scenario, this chapter outlines the various methodologies used for assessing the toxicity of micropollutants on living organisms and human health.