SPSFAM Allergens ERP

AOAC SPSFAM ERP REVIEW: MAIN FORM

Submission Date

2016-09-16 10:45:50

Name

TOMASZ TUZIMSKI

E-mail

tomasz.tuzimski@umlub.pl

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.