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Submission Date
2016-09-16 10:45:50
Name
TOMASZ TUZIMSKI
Organization
MEDICAL UNIVERSITY IN LUBLIN
Title of Method
Detection and Quantitation of Selected Food Allergens using LCMS/MS
AOAC Candidate Method
Number (e.g. ALN-01)
ALL-01
Applicable SMPR
AOAC SMPR 2016.002
Summary:
The proposed method entitled ‘Detection and Quantitation of Selected Food Allergens
using LCMS/MS’ described by Lee Sun New, Hua-Fen Liu, Andre Schreiber, Vincent
Paez is applicable for the detection and quantitation of egg, milk, peanut, and hazelnut
food allergens in finished food products and ingredients by LC-MS/MS.
The method uses triple quadrupole mass spectrometry and selective Multiple Reaction
Monitoring (MRM) of characteristic transitions of precursor ions to fragment ions of
multiple proteins and peptides to uniquely identify each allergen. Characteristic
signature peptides were chosen for each alergen, and MRM transitions for each
signature peptide were determined based on their uniqueness compared to
background proteins and their sensitivity of detection. For each allergen multiple
unique peptides were chosen from unique proteins, and two MRM transitions per
peptide were chosen.
The method performance requirements were met for the detection of egg, milk, peanut
and hazelnut in a number of food matrices.
Food samples were defatted and extracted for proteins. Proteins were digested into
peptides and peptides were separated by LC and detected by MS/MS using multiple,
characteristic MRM transitions on a SCIEX QTRAP® 4500 LC-MS/MS system. The
target analytical range of 5-1000 and 10-1000, respectively, recoveries between 60 and
120% and RSDr % of less than 20% were achieved for all allergens in selected
matrices. RSDR % of less than 30% were achieved for all allergens in bread dough
and bread matrix (three labs with different instruments and operators).
The developed method was evaluated following the definitions of AOAC SMPR
2016.002 with respect to Method quantitation limit (MQL), Method detection limit
(MDL), Linearity, Repeatability, Reproducibility, Recovery. Analytical data was
collected for allergen/matrix combinations listed in AOAC SMPR 2016.002.
In Tables 14 to 17 the authors listed the results for egg, milk, peanut and hazelnut.
Recoveries, RSDr and RSDR are given for the MQL at 5 and 10 ppm, respectively.
RSDr was estimated by analyzing batches in triplicates.
Few allergen-matrices combinations were only analyzed in duplicates RSDr not be
given for these.
The authors are convinced that the method can be easily extended to the detection of
other allergens, including soy and other tree nuts (almonds, Brazil nut, cashew, pine
nut, pistachio, pecan, and walnut).
The authors also declared that an application of this method to matrices not covered
by the scope will require additional validation.
1. Does the applicability of
the method support the
applicability of the SMPR? If
not, please explain what is
missing.
YES: The applicability of the method is adequate to the applicability of the SMPR.
AOAC SPSFAM ERP REVIEW: MAIN FORM