AOAC Guidance on FA Immunoassay Validation (August 2023)

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for each test material (i.e., at each concentration), by each of the three test kit

lots.

5.2.5.2.2.5. Alternatively, method developers may provide internal lot-to-lot and stability

data for review .

5.2.5.3. Data Analysis and Reporting

5.2.5.3.1. Results should be analyzed to determine mean results, repeatability standard

deviation, and recovery for each treatment group.

5.2.5.3.2. Depending on the study type and study design, appropriate statistical tests should be used to whether any experimental conditions or test kit lot characteristics result in

significant differences in method accuracy or precision.

5.2.5.3.3. For the robustness study, data should be analyzed by ANOVA, multi-factor regression,

or generalized linear model software. (11)

5.2.5.3.4. For a product stability and consistency study conducted separately from the matrix study, data should be analyzed by ANOVA. If product stability and consistency are included in a nested design for the matrix study, data should be analyzed according

to the ANOVA procedure outlined in Annex C.

5.2.5.4. Acceptance Criteria

5.2.5.4.1. With respect to robustness study, if any of the experimental conditions evaluated significantly affect the results, this should be mentioned in the kit insert

information.

5.2.5.4.2. For the product stability and consistency study, there should be no significant (p< 0.05) differences between test kit lots and no significant effect of storage time.

5.3. Independent Laboratory Study (PTM)

5.3.1. Scope 685 The independent laboratory validation study, conducted under PTM program guidance, should verify 686 the analytical results obtained in the method developer study in a controlled laboratory setting. The 687 independent laboratory should verify the precision (repeatability and intermediate precision), 688 sensitivity, and recovery performance parameters of the method. 689 5.3.2. Matrix Study 690 5.3.2.1. Test Materials 691 5.3.2.1.1. Incurred test materials are required for evaluation of precision, sensitivity, and 692 recovery. See Annex A for description of best practices for incurred test material 693 preparation. 694 5.3.2.1.2. At minimum, the independent laboratory must analyze at least one food matrix for 695 every five matrices evaluated in the Method Developer Study. Food matrices do not 696 include cleaning and sanitation solutions such as CIP rinse water. The independent 697 laboratory must analyze at least one environmental surface/cleaning and sanitizing 698 solution for every five environmental surfaces/cleaning and sanitizing solutions 699 claimed. The selection of which matrices/surfaces/cleaning and sanitizing solutions 700 are analyzed should be reflective of the range of difficulty associated with the 701 claimed matrices. 702