AOAC Guidance on FA Immunoassay Validation (August 2023)

1. Scope 62 The purpose of this document is to provide AOAC International (AOAC) guidelines for method developers 63 conducting validation studies of immunoassay-based food allergen analysis methods, e.g., for 64 submission for AOAC Official Methods of Analysis (OMA) status and/or Performance Tested Methods 65 (PTM) certification. This document is not intended to describe requirements for laboratories using 66 commercial immunoassay methods for food allergen analysis, though it may assist these laboratories in 67 understanding the consensus-based validation approaches, terminology used, and information expected 68 to be provided by method developers. 69 2. Applicability 70 These guidelines are applicable to both quantitative and qualitative food allergen methods, of either a 71 proprietary or non-proprietary nature. 72 3. Terms and Definitions 73 3.1. General 74 3.1.1. Analyte: 75 3.1.1.1. chemical entity or entities* measured by the measurement system (see also ‘measurand’ 76 and narrative discussion). *which may be a marker(s) or surrogate(s), see Section 4.1.4 77 3.1.1.2. See also “measurand” definition and narrative examples (Section 4.1.4). See De Bievre 2013 78 for a detailed discussion of the difference between “analyte” and “measurand”.(1) 79 3.1.2. Enzyme-linked immunosorbent assay (ELISA): an analytical procedure characterized by the 80 recognition and binding of specific antigens by antibodies and signal generation by an enzyme- 81 substrate reaction 82 3.1.3. Food allergen: A food source known to have the ability to elicit IgE-mediated allergic reactions in 83 susceptible individuals, generally identified by the common name of the food, e.g., milk, egg, fish, 84 crustacean shellfish, soy, peanut 85 3.1.4. Food allergen material: A food ingredient or component derived from a food allergen 86 3.1.5. Matrix: Totality of components of a material system except the analyte (ISO 17511). For example, 87 the food, beverage, or environmental surface material to be included in the validation as per the 88 intended use of the method. 89 3.1.6. Measurand: The quantity intended to be measured (the specification of the measurand should be 90 sufficiently detailed to avoid any ambiguity). See also “analyte” definition and narrative examples 91 (Section 4.1.4) 92 3.1.7. Test material: A material used for method validation that either contains a target food allergen 93 present at a given concentration in the context of a food or environmental matrix or is a blank 94 matrix free of the target food allergen. 95 3.1.8. Spiked Test Material: A food matrix into which a food allergen material has been incorporated 96 after all relevant food processing operations have been completed (See Annex A for details). 97 3.1.9. Incurred Test Material : Prepared from a food matrix into which a food allergen material has been 98 incorporated prior to subjecting the matrix to a given food processing operation. 99 3.1.10. Qualitative method: Method of analysis whose response is either the presence or absence of the 100 analyte. 101