Development of a simplified, sensitive, and accurate LC-MS/MS method for peanut quantification in wheat flour-based...

Area of Science:

• Food Science
• Analytical Chemistry
• Allergen Detection

Background:

• Enzyme-linked immunosorbent assay (ELISA) methods can overestimate peanut quantification in wheat flour matrices.
• Accurate detection of peanut allergens is crucial for food safety and consumer protection.

Purpose of the Study:

• To develop and validate a reliable LC-MS/MS method for accurate peanut quantification in wheat flour.
• To compare the performance of the LC-MS/MS method with ELISA kits.

Main Methods:

• Developed a simplified sample preparation protocol involving protein digestion without reduction and alkylation.
• Utilized liquid chromatography tandem mass spectrometry (LC-MS/MS) for precise peanut peptide detection.
• Employed Ara h 1 for calibration curve construction and analyzed heated extracts to enhance peptide detection.

Main Results:

• The LC-MS/MS method demonstrated superior sensitivity and linearity compared to commercial ELISA kits.
• Achieved a limit of quantification (LOQ) of 0.31 mg/kg and a linear range of 0.3–40 mg/kg for total peanut in wheat flour.
• Heating extracts significantly increased the detection of peanut peptides.

Conclusions:

• The developed LC-MS/MS method provides accurate and reliable quantification of peanut in wheat flour.
• This method serves as a valuable confirmatory tool for peanut allergen management, especially when ELISA results are questionable.