Research project
36 | monthsIMPROVING

Multimethodological approach to evaluate chemical-biological and toxicological hazards

Related toSpoke 03

Principal investigators
Maria Elisa Crestoni,Alessandra Gentili,Cinzia Ingallina,Massimo Reverberi

Other partecipantsAntonella Di Sotto, Silvia Di Giacomo, Marco Ruggeri, Luigi Faino
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Highlights

Task involved

Task 3.1.1.

Standard protocols (ISO), whole genome sequencing (WGS), computational methodologies, and MetaOmic approaches (metagenomics, metatrascriptomics, metabolomics, lipidomics, culturomics and phenomics) will be applied for the identification and characterization of the new and (re)-emerging chemical and biological hazards in traditional products, related to climate changes, microbial evolution, and modifications in the manufacturing processes. Omics techniques will also be applied to study factors affecting the survival and the stress resistance mechanisms of pathogens and antimicrobial resistant (AMR) bacteria during food processing and shelf life. In addition, a CAD-based automatic feature recognition procedure will be developed for hygienic design of food machinery, as a prerequisite for GMP in food production.

Task 3.1.2.

MetaOmics and MultiOmics approaches will be used for the identification and quantification of toxic compounds in novel sources or ingredients, also to anticipate risks related to novel foods and new sustainable food processes. The culture collection from task 3.1.1 will be used for challenge experiments in pilot plants mimicking industrial manufacturing. Food toxicants in new ingredients and food processing technologies will be evaluated (in connection with Spoke 2). Allergenicity (also with computational approaches) and toxicity will be considered by in vitro and in vivo tests. Exposure assessment and risk- benefit assessment (RBA) of novel foods will be performed.

Task 3.1.3.

New/existing materials will be characterised in terms of migration studies of both intentionally and non-intentionally added substances, small-/micro/nano- plastics, metal nanoparticles, food packaging suitability and growth of mycotoxigenic moulds, even after being subjected to innovative/emerging processing technologies (e.g., cold gas plasma, HPP). Potential antimicrobial properties will also be assessed, particularly for developed functional packaging systems. The safety of new materials used as FCM, from recyclable sources, bioplastics or derived from by-products, will be assessed in collaboration with Spoke 2

Project deliverables

D3.1.1.2.

Omics protocols to evaluate food safety

D3.1.1.3.

Report on RA of Italian traditional foods (M36)

D3.1.3.1.

Report on safety / stability functionality of new/existing food packaging materials/systems (M30)

D3.1.2.2.

RA of food toxicants (M30)

D3.1.2.3.

Report on integrated methodologies for RA and RBA of novel foods (M30)

D3.1.2.4.

Report on the formation, accumulation, and modification of food toxicants along the food production chain

State of the art

The food and beverage production is constantly expanding and a comprehensive quality control system is required to monitor each step of the chain, from production to retail, ensure environmental and human safety and improve consumer’s health and confidence. 

Chemical and metabolic transformations deriving from the effects of (new) manipulations, storage and packaging conditions may largely affect the composition of traditional and novel foods, of plant and animal origin. 

A major effort involves developing analytical protocols to maximize metabolome coverage and identify (potentially) harmful compounds from sanitary and production treatments.   

Here, a comprehensive and high-throughput bioanalytical strategy is proposed based on high-resolution NMR and MS techniques, and biochemical assays to explore the chemical diversity of different food matrices, the presence of contaminants and hazards of natural and anthropogenic origin, and to determine the molecular effects of processing and storage on food safety and genuineness.

Operation plan

Food matrices throughout spoke 3 and in collaboration with spoke 2 and 4 will be listed and classified. Among them, pilot products will be selected along the production and transformation process for the application of a multi-methodological approach, developed following the three parallel lines:

  • Chemical characterization
    Untargeted NMR and MS-based metabolomic methods and targeted chromatographic and spectrophotometric methods (HPLC-PDA, HPLC-MS/MS, HPLC-UV/FD, UV-Vis spectrophotometer) will be applied for the identification of chemico-biological hazards related to phytosanitary treatments, manufacturing processes, incorrect storage practices and (a)biotic stressors.
  • Targeted sequencing
    Identification of biohazard agents in fresh food using targeted analysis. Detection of biological agents like Escherichia coli or Salmonella with Polymerase chain reaction.
  • Toxicological characterization
    Identification of hazards arising from toxic compounds, contaminants, and impurities. Determination of cytotoxicity in human cell models of gastro-intestinal tract by spectrophotometric, fluorimetric and cell imaging methodologies. Genotoxicity assessment according to OECD guidelines.

Expected results

Highly complementary, advanced NMR and MS techniques will be applied to provide a broad chemical and metabolomic profile and monitor traceability and quality of traditional, novel, and transgenic (GM) foods (in collaboration with spoke 2 and 4). 

The target is also related to the development of a widely applicable methodology to identify possible sources of human disease in food. Food processing and shelf-life will also be evaluated. While NMR supports molecular structure characterization and absolute quantification of detected signals, the huge dynamic range and sensitivity of MS techniques enables harmful compounds present in trace amounts, like pesticides, agrochemical derivatives, metal and metalloids containing complexes, and antiseptics, to be determined in very heterogeneous and complex matrices. Peptides/Protein identification for food profiling and quality will be acquired to point out adulteration or aggressive industrial practices.