ABSTRACT
In this study, an ELISA-based method was observed to overestimate quantification of peanut in three wheat flour-based dry matrices (both raw and cooked). A reliable method using liquid chromatography tandem mass spectrometry (LC-MS/MS) was developed to quantify peanut accurately in these matrices. This method simplified both sample preparation and quantification compared with previous approaches, by digesting peanut proteins without reduction and alkylation and using Ara h 1 to build calibration curves. A significantly increased amount of peanut peptides was detected by heating the extracts. The sensitivity and linearity were superior to most commercial ELISA kits. The limit of quantification (LOQ) was 0.31 mg/kg total peanut in wheat flour and the linear range was between 0.3 and 40 mg/kg. This LC-MS/MS method could be applied as a confirmatory method for peanut allergen management when positive ELISA results raise suspicion of contamination.
Acknowledgments
The authors would like to thank Agilent Technologies (Jerry Zweigenbaum, Linfeng Wu) for technical consultation on sample preparation. The authors would also like to thank John Carney and Liesbeth Bruckers for their advice on statistical analysis. The authors would like to dedicate this paper to Mr. David Crean for his 35 years of stellar contribution to Mars.
Disclosure statement
No potential conflict of interest was reported by the author(s).
Supplementary material
Supplemental data for this article can be accessed on the publisher’s website.