ISPAM Working Group Chair Presentation Book

Challenges

• Unique protein and peptide selections with different digestion conditions  – Perform protein ID for allergens: Trypsin; chymotrypsin; species selected based on the  list highlighted by The Codex Alimentarius. – Search against database and select unique proteins/peptides.   Goal : • Detection of two proteins each species (if available), 3‐4 peptides each protein, and 3‐4 MRM  transitions each peptide. • MRM transition selections with consideration on sensitivity and few/no interferences from  the food matrix. • MRM transition selections made with consideration of potential modification during food  processing (cooking, baking, fermenting etc. )and storage (i.e. MRMs must be detectable in  raw and processed food) • Sample clean up: lipids removal   – Try different solvents (ex. hexane, etc) • Protein extraction and digestion optimization  – Try different denature conditions: (ex. heat, urea, OGS, guanidine).  – Final combined mixture of denature reagent.   – Must be simple for sample preparation and easy to reproduce.