Incorporation of pea (Pisum sativum), bean (Phaseolus vulgaris) and lentil (Lens culinaris) by-product protein extracts into food matrices: A safety perspective

The rising demand for plant-based meat-like products has led to increased interest in legume proteins, while significant portions of by-products are discarded in the EU. This study explored pea (Pisum sativum), bean (Phaseolus vulgaris), and lentil (Lens culinaris) protein extracts as viable food ingredients. Safety assessments were performed by heavy metal detection, microbiological analysis and pesticide assessment. Additionally, in vitro assays were conducted to assess cytotoxicity using the MTT and Alamar Blue assays in HepG2 cells, while genotoxicity and mutagenicity were evaluated using the Comet assay and Ames test, respectively. Results showed that heavy metal concentrations in all extracts were below Codex Alimentarius limits. The extracts showed no contamination by pathogenic microorganisms as well. Pesticide residues in all raw material samples were below the EU's maximum residue level (MRL). In vitro cytotoxicity assays indicated minimal impact on HepG2 cell viability, while genotoxicity and mutagenicity tests confirmed no DNA damage or mutagenic potential. These findings support the safety of pea, bean, and lentil by-product protein extracts for use in food formulations. Furthermore, this study highlights the importance of New Approach Methodologies (NAMs) in food safety assessments, reducing reliance on animal testing while ensuring consumer protection.