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

Importance of the Topic

Food allergies pose significant health risks worldwide, and stringent labeling regulations demand reliable methods to detect trace levels of allergenic proteins in complex food matrices. Traditional techniques such as ELISA and PCR face limitations in multiplexing capability and specificity, motivating the development of advanced analytical approaches.

Objectives and Study Overview

This work aimed to establish a rapid, sensitive, and selective method for the simultaneous quantification of eight priority allergens (wheat, buckwheat, egg, milk, peanut, shrimp, crab, and soy) in processed foods. The study outlines sample preparation, chromatographic separation, and mass spectrometric detection strategies to achieve robust multiplex analysis.

Used Instrumentation

• NexeraTM X3 high-performance liquid chromatograph
• Shimadzu LCMS-8060NX triple quadrupole mass spectrometer

Methodology

Standard extracts of target allergenic proteins were subjected to reduction, alkylation, and trypsin digestion, followed by solid-phase extraction cleanup. The resulting peptide mixtures were separated on reversed-phase HPLC under a 10.5-minute gradient and analyzed via multiple reaction monitoring (MRM). A total of 48 MRM transitions targeting 17 signature peptides enabled concurrent detection with a calibration range of 0.1–50 ppm.

Main Results and Discussion

All eight allergenic peptides exhibited sharp, well-resolved peaks within the chromatographic run time. Calibration curves demonstrated excellent linearity and sensitivity, exemplified by the crustacean peptide AGGLTLER. Application to commercial food samples showed no false positives in pre-packaged curry or baby food, while wheat peptides were correctly identified in udon noodles, confirming alignment with product labels.

Benefits and Practical Applications

• Enables simultaneous monitoring of multiple allergens in a single analysis
• Reduces false positives and improves specificity compared to immunoassays
• Short run time enhances laboratory throughput for QA/QC and regulatory testing

Future Trends and Potential Applications

Advancements may include expanding the peptide panel to cover additional allergens, integrating high-resolution mass spectrometry for enhanced specificity, automating sample preparation workflows, and deploying real-time monitoring solutions in food production environments. Such developments will further strengthen food safety and compliance assurance.

Conclusion

A robust LC-MS/MS method using triple quadrupole instrumentation was successfully developed for the simultaneous detection of eight key food allergens. Demonstrated accuracy and speed in real food matrices underscore its value for routine allergen screening in food quality control laboratories.