The Growing Global Demand for Sustainable and Alternative Protein Sources
The growing global demand for sustainable and alternative protein sources has increased interest in edible insects. Their regulatory approval in the European Union as novel food and feed ingredients necessitates reliable analytical methods to ensure food safety, authenticity, and allergen risk assessment. This thesis addresses these challenges by developing and validating mass spectrometry-based proteomic workflows for insect protein identification, authentication, and quantification. The research establishes novel methodologies to support food safety, traceability, and regulatory compliance in the emerging insect protein industry.
Challenges in Insect Protein Analysis
Lack of comprehensive, species-specific protein databases limits proteomic analysis
High variability in protein composition across insect species poses identification challenges
Allergenic potential requires thorough assessment to ensure consumer safety
Regulatory compliance demands accurate and reliable detection methods
Direct proteolysis and sample cleanup are critical for efficient analysis
Novel Proteomic Workflows for Insect Proteins
A homology-based nano-HPLC-HR-MS approach was developed to overcome database limitations
Up to 1893 proteins were identified across six insect species, representing a 24-fold increase in available proteomic data
Parallel reaction monitoring confirmed 90% of tested peptides, validating the workflow
Species-specific marker peptides and a conserved pan-insect tropomyosin marker were identified
Conserved allergens like tropomyosin were detected across all tested insect species
Species-specific allergens indicated unique allergenic risks for different species
Targeted immunoaffinity LC-MS/MS assays were developed for authentication and quantification
The assays demonstrated high accuracy, precision, selectivity, and sensitivity
Reliable detection of insect proteins in commercial products and model food matrices was achieved
The thesis successfully develops and applies novel analytical tools for the comprehensive proteomic characterization of non-sequenced insect species. The methodologies enable both qualitative and quantitative detection of insect proteins in food, supporting food safety, traceability, and regulatory compliance. This research provides a robust framework for the safe and sustainable integration of insect proteins into the global food supply, addressing key challenges in authentication, allergen risk assessment, and quality control.
