AOAC SPSFAM ERP for SELECTED FOOD ALLERGENS

AOAC SPDS ERP – ALL-01 Review

Name

Melanie Downs

E-mail

mdowns2@unl.edu

Organization

University of Nebraska-Lincoln

Detection and Quantification of Selected Food Allergens using LC-MS/MS (Revision 2)

Title of Method

AOAC Candidate Method Number (e.g. ALN-01)

ALL-01

Applicable SMPR

2016.002

The method described in the submission can broadly be described as a targeted bottom-up proteomics detection method. The authors used discovery bottom-up proteomics to identify target tryptic peptides from each allergenic food. These target peptides were subsequently incorporated into the final targeted method, using a selected reaction monitoring (SRM) strategy to monitor specific transitions of these peptide targets. The method claims to detect and quantify egg, milk, peanut, and hazelnut in several different matrices. The sample preparation described is consistent with a typical bottom-up proteomics experiment, including protein extraction, reduction, alkylation, trypsin digestion, and sample clean-up. Defatting is also an optional step prior to protein extraction for some matrices. In some instances, internal standard peptides are also added to the peptide digests. The digested samples are subsequently separated by RP-HPLC, inline with ESI-MS/MS analysis by a QTRAP instrument. The pre-established peptide transitions were monitored with a scheduled SRM method over the course of a 12 minute

Summary: