In Vitro New Approach Methodologies for Toxicity Assessment of Emerging Food Contaminants in Indonesia: A Proposed Strategy for BPOM Regulatory Integration

ABSTRACT

The accelerating global diversification of food chemicals—encompassing mycotoxins, heavy metals, organophosphate pesticide residues and the increasingly prevalent burden of microplastics and nanoplastics (MNPs)—creates an urgent demand for rapid, human‐relevant and ethically acceptable toxicity testing platforms. Conventional animal‐based bioassays are insufficient to characterise the thousands of substances awaiting safety evaluation, a deficit particularly acute in tropical food‐producing regions including Indonesia where environmental contamination profiles and regulatory infrastructure differ markedly from high‐income countries. New approach methodologies (NAMs), comprising validated in vitro cell and organoid systems, high‐throughput screening (HTS), multiomics toxicogenomics, in silico modelling and adverse outcome pathway (AOP) frameworks, represent a paradigm shift in food safety toxicology. This review critically appraises current evidence on in vitro NAMs for emerging food contaminants, with emphasis on (i) 2D cell line and 3D organoid/organ‐on‐chip models for hepatotoxicity, enterotoxicity and nephrotoxicity; (ii) mechanistic pathways—oxidative stress, mitochondrial dysfunction, genotoxicity and intestinal barrier disruption—as interrogated through standardised in vitro endpoints; (iii) AOP frameworks as regulatory bridges between molecular initiating events and adverse outcomes; (iv) high‐throughput transcriptomics (HTTr) and Cell Painting integration for mechanism‐informed hazard prioritisation; and (v) in vitro–to–in vivo extrapolation (IVIVE) via physiologically based toxicokinetic (PBTK) modelling. Critical gaps in chronic‐exposure data, mixture toxicity assessment and BPOM regulatory acceptance are discussed. A novel evidence‐based NAM validation roadmap tailored to the Southeast Asian regulatory context is proposed.