Unraveling microplastic behavior in simulated digestion: Methods, insights, and standardization
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Updated: May 2, 2026

Protocol for Microplastics Sampling on the Sea Surface and Sample Analysis
Published on: December 16, 2016
Gurusamy Kutralam-Muniasamy1, V C Shruti2, Fermín Pérez-Guevara3
1Department of Biotechnology and Bioengineering, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Av Instituto Politécnico Nacional 2508, San Pedro Zacatenco, Gustavo A. Madero, Ciudad de México 07360, Mexico; CIITEC - IPN. Centro de Investigación e Innovación Tecnológica, Cda. de Cecati s/n, Santa Catarina, Azcapotzalco, Ciudad de México 02250, Mexico.
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.
Insights
In vitro microplastic (MP) digestion studies lack standardization, hindering comparisons and risk assessments. This review synthesizes 85 studies, highlighting methodological inconsistencies and the need for unified protocols to understand MP health risks.
Area of Science:
- Environmental Science
- Toxicology
- Analytical Chemistry
Background:
- In vitro digestion studies on microplastics (MPs) are rapidly expanding but lack standardized methodologies.
- Inconsistent protocols hinder cross-study comparisons, risk assessments, and understanding MP fate in the gastrointestinal tract.
- A comprehensive synthesis is needed to address fragmentation and establish a unified research direction.
Purpose of the Study:
- To systematically evaluate 85 in vitro digestion studies on microplastics published between 2020 and 2024.
- To consolidate key findings, identify methodological challenges, and assess disparities in digestion protocols.
- To provide a foundation for improving reproducibility and advancing standardization efforts in microplastic research.
Main Methods:
- Systematic literature review of 85 studies (2020-2024) on in vitro microplastic digestion.
- Analysis of variations in digestion protocols, fluid compositions, and exposure conditions (pH, enzymes, temperature, time).
- Synthesis of findings on bio-corona formation, structural changes, contaminant bioaccessibility, and enzyme interactions.
Main Results:
- Significant disparities exist in digestion protocols, fluid compositions, and exposure conditions across studies.
- Microplastic behavior and physicochemical transformations are heavily influenced by factors like pH, enzyme activity, and residence time.
- Bio-corona formation, structural modifications, and contaminant bioaccessibility vary based on experimental conditions.
Conclusions:
- The field of in vitro microplastic digestion is fragmented due to methodological inconsistencies, particularly in human-based models.
- There is an urgent need for standardized methodologies to improve reproducibility and enable reliable risk assessments.
- Addressing these gaps will enhance understanding of microplastic ingestion risks to human health.
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