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AOAC SPDS ERP – ALL-01 Review
Name
Melanie Downs
Organization
University of Nebraska-Lincoln
Title of Method
Detection and Quantification of Selected Food
Allergens using LC-MS/MS (Revision 2)
AOAC Candidate Method
Number (e.g. ALN-01)
ALL-01
Applicable SMPR
2016.002
Summary:
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