AOAC Gluten Qualitative Validation Guidance-FINAL (July 2023)

Q UALITATIVE G LUTEN M ETHOD V ALIDATION G UIDELINES PG . 3

and 0.5M NaCl (CODEX STAN 118-1979). Throughout this document, the word ‘wheat’ refers to all Triticum species, such as durum wheat, spelt and khorasan wheat, and their hybrids and crossbred varieties such as Triticale. 3.11 High-Dose Hook Effect The result of a suboptimal antigen-antibody reaction in which either the antibody or antigen is in excess, incomplete, or blocks an optimal reaction (CLSI EP12-A2). Also known as Prozone Effect or Overload Effect. 3.12 Incurred Test Material A test material prepared from a food matrix into which a gluten source (e.g. flour) has been incorporated prior to subjecting the matrix to a given food processing operation. 3.13 Independent Testing Site A testing site not owned, operated or controlled by the same entity as the method developer. 3.14 Interference Study The examination of matrices expected to be tested with the method, to demonstrate that they do not interfere with detection of the analyte. 3.15 Lateral Flow Device (LFD) An analytical method characterized by use of an immunochromatography platform for the detection of specific analyte. 3.16 LPOD Composite POD pooled across laboratories. Includes between-laboratory variation in addition to variation inherent in the binomial mature of binary probabilities (INSERT UPCOMING JAOAC REF). 3.17 Matrix The totality of components of a material system except the analyte (ISO 17511). For example, the food, beverage, or environmental surface material to be included in the validation as per the intended use of the method. 3.18 Measurand The quantity intended to be measured (the specification of the measurand should be sufficiently detailed to avoid any ambiguity). See also Analyte . 3.19 Measurement interference: A cause of significant bias in the measured analyte concentration due to the effect of another component or property of the sample which may result from non specificity of the detection system, suppression of an

Definitions

3.1 Analyte Chemical / molecular entity or entities, which may be surrogate(s), measured by the measurement system (see also Measurand ). 3.2 Candidate Method The method submitted for validation 3.3 Candidate Method Result The final result of the qualitative analysis for the candidate method. 3.4 Claimed Detection Capability (CDC) An analyte concentration, expressed in mg/kg of gluten (or µ g per surface area), that demonstrates a Probability of Detection (POD) of at least 0.95. This may be claimed as one level that achieves a POD of 0.95 across all validated matrices, or individually per matrix and/or gluten source. The claimed detection capability must be verified empirically during method validation. 3.5 Collaborator An intended user who participates in the collaborative study. 3.6 Cross-reactivity A measurable positive response, above the LOD or CDC of the method, to a material other than the target analyte. 3.7 Cross-reactivity Study The examination of matrices that do not contain the claimed analyte, which are potentially cross-reactive, to determine that they are not detected by the method. 3.8 Data Set Results from a combination of an analytical instrument, device, or equipment and an operator, technician or analyst. Results from a single test site may serve as multiple data sets dependent on nonredundant operators and acceptable separation of effort. 3.9 Fractional POD Range Validation criterion that is satisfied when a test material yields both positive and negative responses within a set of replicate analyses. See also POD. 3.10 Gluten A protein fraction from wheat, rye, barley, oats 1 or their crossbred varieties and derivatives thereof, to which some persons are intolerant and that is insoluble in water

1 Oats can be tolerated by most but not all people who are intolerant to gluten. Therefore, the allowance of oats that are not contaminated with wheat, rye or barley in foods […] may be determined at the national level (CODEX STAN 118-1979)