A key component of ecological risk assessments is to develop evidence-based benchmarks to assess potential hazards to various receptors. To ensure that toxicity value development is performed using the best available science, the reliability (or inherent scientific quality) of these studies must be considered. The degree of reliability can be evaluated via critical appraisal tools (CATs), though application of such methods assessing ecotoxicological literature for toxicity value development is not well established compared to human health assessments. A review of existing CATs revealed that there is currently no approach that considers the full range of biases that should be considered for appraisal of internal validity in ecotoxicological studies. Recognizing the need for a comprehensive ecotoxicity study assessment framework that addresses risk of bias (RoB) for the interpretation of study reliability, we propose the Ecotoxicological Study Reliability (EcoSR) framework. This framework builds upon the classic RoB assessment approach, frequently applied in human health assessments. EcoSR adds reliability and key criteria specific to ecotoxicity studies from existing appraisal methods, emphasizing those used by regulatory bodies. Additionally, the framework recommends a priori customization based on assessment goals. EcoSR is comprised of two tiers: an optional preliminary screening (Tier 1) and a full reliability assessment (Tier 2). The framework outlines a systematic approach for conducting ecotoxicity study appraisals that enhances transparency and consistency in determining study reliability. The EcoSR framework provides flexibility and can be refined and applied to a variety of chemical classes. This framework represents a significant step towards improving the transparency and reproducibility of ecotoxicological study appraisals; ultimately, contributing to more informed and reliable toxicity value development within the ecological sciences.