Simultaneous analysis of major allergens in food matrices by high sensitive mass spectrometer

Význam tématu

Food allergies pose significant health risks and concern for both consumers and food manufacturers. Proteins in foods such as milk, eggs, fish, shellfish, tree nuts, peanuts, wheat and soybeans can trigger severe immune reactions. Reliable detection of these allergenic proteins is critical for consumer safety, accurate labeling and compliance with regulations such as FALCPA. Traditional assays like ELISA and PCR have limitations in specificity and multiplexing, driving adoption of LC-MS/MS for sensitive and selective simultaneous analysis of multiple allergens.

Cíle a přehled studie

This study aimed to develop a high-throughput LC-MS/MS method for the simultaneous detection of major food allergens. The approach targeted signature peptides from 13 proteins across eight allergenic food groups. Method performance was evaluated in both standard mixtures and real food matrices, including cooked and gluten-free products, to demonstrate applicability in complex samples.

Použitá metodika a instrumentace

• Sample preparation: Milling of food samples followed by defatting with hexane, protein extraction using Tris-HCl buffer with urea and protease inhibitors, reduction, alkylation and in-solution digestion.
• Peptide cleanup: Solid phase extraction and lyophilization.
• LC-MS/MS: Shimadzu Nexera X2 UHPLC coupled to LCMS-8050 triple quadrupole mass spectrometer.
• Chromatography: Shim-pack XR-ODS III column, gradient elution with formic acid in water and acetonitrile over a 10 min run.
• MRM transitions: 150 optimized transitions for 31 signature peptides using Skyline for transition selection and collision energy optimization.

Hlavní výsledky a diskuse

• All target peptides were baseline separated within 6.5 minutes, showing excellent linearity (R2 >0.999) over 1–50 ppm.
• Detection of all eight allergenic food proteins in mixed standard matrices and various cooked food samples, with reliable identification of labeled allergens in commercial products.
• Gluten peptides were absent in gluten-free bread and crackers, and wheat peptides were detectable at spiking levels as low as 20 ppm.
• Selection of unique peptide sequences minimized false positives from related grains, despite peptide homology in certain cereals.

Přínosy a praktické využití metody

The developed LC-MS/MS method enables rapid, multiplexed detection of major food allergens with high sensitivity and specificity. It supports regulatory compliance for allergen labeling and can be integrated into quality assurance workflows in food manufacturing. The approach overcomes cross-reactivity issues of immunoassays and expands analytical capabilities beyond single-analyte tests.

Budoucí trendy a možnosti využití

• Expansion of quantitative capabilities by incorporating stable isotope labeled peptides.
• Development of targeted methods for less common allergens and emerging allergenic proteins.
• Integration with automation and high-throughput sample preparation to increase laboratory throughput.
• Application of this platform for routine monitoring and risk assessment in supply chains and processed foods.

Závěr

A robust LC-MS/MS workflow has been established for comprehensive analysis of major food allergens in various matrices. The method demonstrates rapid analysis time, high sensitivity and selectivity, and practical applicability for ensuring food safety and regulatory compliance.

Použitá instrumentace

• Shimadzu Nexera X2 UHPLC system
• Shimadzu LCMS-8050 triple quadrupole mass spectrometer
• GM-200 Retsch grinder