Unraveling microplastic behavior in simulated digestion: Methods, insights, and standardization

Abstract

Despite the rapid expansion of in vitro digestion studies on microplastics (MPs), the field remains fragmented due to inconsistent methodologies, varying analytical approaches, and a lack of standardized protocols. These discrepancies hinder cross-study comparisons, complicate risk assessments, and limit the applicability of in vitro models for understanding MP fate and pollutant interactions in the gastrointestinal environment. A comprehensive synthesis is needed to assess progress, identify research gaps, and establish a unified research direction. This review systematically evaluates 85 studies (2020–2024), consolidating key findings and methodological challenges. It examines disparities in digestion protocols, fluid compositions, and exposure conditions, assessing how factors such as pH, enzyme activity, residence time, and temperature shape MPs' behavior and physicochemical transformations. Key findings on bio-corona formation, structural modifications, contaminant bioaccessibility, and interactions with digestive enzymes are synthesized to provide a clearer picture of MP behavior during digestion. With the field remains dominated by studies on polystyrene and polyethylene MPs in human-based models, inconsistencies persist, highlighting the urgent need for standardized methodologies. By addressing these gaps, this review lays a critical foundation for improving reproducibility, advancing standardization efforts, and strengthening exposure assessments, ultimately enhancing our understanding of MP ingestion risks to human health.