Advancing High-Throughput MS-Based Protein Quantification: A Case Study on Quantifying 10 Major Food Allergens by...

Area of Science:

• Analytical Chemistry
• Food Science
• Mass Spectrometry

Background:

• Liquid chromatography-mass spectrometry (LC-MS) is the standard for food allergen quantification.
• Traditional calibration curves (MSCC) are time-consuming and prone to errors.
• There is a need for more efficient and reliable methods for food allergen analysis.

Purpose of the Study:

• To develop and validate a sensitive and robust LC-MS/MS method for quantifying 10 major food allergens.
• To replace the traditional calibration curve method with a more efficient one-sample multipoint external calibration curve (OSCC).
• To utilize the multiple isotopologue reaction monitoring (MIRM) technique for improved calibration curve establishment.

Main Methods:

• Development of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method.
• Implementation of a one-sample multipoint external calibration curve (OSCC) strategy.
• Application of the multiple isotopologue reaction monitoring (MIRM) technique with a single spiked level in a blank sample.
• Validation of the method for selectivity, sensitivity, accuracy, and precision.

Main Results:

• The proposed LC-MS/MS method with OSCC and MIRM demonstrated excellent performance.
• Method selectivity, sensitivity, accuracy, and precision were comparable to traditional MSCC.
• The strategy enabled isotope sample dilution by monitoring less abundant MIRM channels.
• The method was successfully applied to detect 10 food allergens in commercial food products.

Conclusions:

• The MIRM-OSCC-LC-MS/MS methodology offers a high-throughput and robust alternative for food allergen quantification.
• This approach simplifies sample preparation and reduces errors associated with traditional calibration curves.
• The developed method has significant potential for MS-based protein quantification analysis, particularly in food safety and quality control.