Development of a Simultaneous Analysis Method for Allergens in Food Using a Triple Quadrupole Mass Spectrometer

Importance of the Topic

Food allergies are triggered by immune responses to specific proteins in food and pose significant health risks. Reliable detection of allergenic ingredients is essential for consumer safety and regulatory compliance in the food industry. Traditional methods such as ELISA and PCR have limitations in specificity, cross-reactivity and multiplexing capacity, creating demand for high-selectivity, high-sensitivity approaches.

Objectives and Study Overview

The study aimed to develop a rapid, simultaneous analysis method for seven specified allergens (wheat, buckwheat, eggs, milk, peanuts, shrimp, crab) and one equivalent allergen (soy) in processed foods. The goal was to achieve clear peptide separation, robust quantitation and accurate detection consistent with label claims in commercial products.

Methodology and Used Instrumentation

Sample Preparation Workflow

• Protein extraction from samples
• Reduction of disulfide bonds and cysteine alkylation
• Trypsin digestion to generate target peptides
• SPE purification and concentration

Chromatography and Mass Spectrometry

• UHPLC: Nexera X3 system with Shim-pack GIST-HP C18-AQ column (2.1×100 mm, 1.9 μm)
• Mobile phases: 0.1 % formic acid in water (A) and acetonitrile (B); gradient from 2 % to 95 % B over 13.5 min
• Triple quadrupole MS: LCMS-8060NX with IonFocus ESI in positive MRM mode
• MRM transitions: 48 transitions targeting 17 peptides across eight allergens

Main Results and Discussion

All target peptides eluted within 10.5 minutes with sharp, well-resolved peaks. Linearity was demonstrated from 0.1 to 50 ppm for key peptides, yielding R2 values above 0.996. Analysis of pre-packaged curry, baby food and udon noodles showed no allergen signals when none were labeled, and selective detection of wheat in noodles. Spiked samples at 2.5 ppm and 10 ppm confirmed accurate recovery and absence of false positives.

Benefits and Practical Applications

This LC-MS/MS method enables simultaneous quantification of multiple food allergens in processed products, reducing analysis time and resource use compared to separate ELISA or PCR assays. Its high specificity avoids cross-reactivity and DNA-based ambiguities, supporting quality control in food manufacturing and regulatory testing.

Future Trends and Potential Applications

Advances may include expanded allergen panels, high-throughput multiplexing, and integration with automated sample prep. Coupling with data analytics and real-time monitoring could further enhance allergen surveillance and compliance in complex food matrices.

Conclusion

A robust LC-MS/MS workflow using Nexera X3 and LCMS-8060NX was developed for simultaneous analysis of eight food allergens. The method delivers rapid separation, high sensitivity and reliable quantitation, proving suitable for practical use in food safety laboratories.