AOAC Guidance on FA Immunoassay Validation (August 2023)

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5.2.4.3.3.1. Percent Recovery = (Experimental concentration)/(Expected concentration) x

100

5.2.4.3.3.2. The expected concentration for each test material should be calculated from

the incurred concentration, accounting for any mass changes during processing operations (e.g., moisture loss during baking).

5.2.4.4. Acceptance Criteria

5.2.4.4.1. If an applicable SMPR is available for a method, the SLV study data must meet the

corresponding criteria.

5.2.4.4.2. In the absence of an applicable SMPR, an expert review panel will evaluate the study data according to their expert opinions. With respect to recovery, while ideal mean recovery values are from 80-120%, values of 50-150% are acceptable. (9)

5.2.4.4.3. LOQ

5.2.4.4.3.1. The LOQ must be greater than or equal to the LOD.

5.2.4.4.3.2. The RSD i at the LOQ must be less than or equal to the RSD i in the relevant SMPR (or the RSD R if an RSD i is not listed). If there is no SMPR available for a

particular food allergen, RSD i at the LOQ must be ≤ 30%.

5.2.4.4.3.3. If a method developer has an LOQ claimed as part of the method design (e.g., the lowest non-zero calibrant), the estimated LOQ from the SLV (which meets the SMPR requirements for maximum RSD i ) must be less than or equal to the claimed LOQ of the kit, within statistical tolerances. If the estimated LOQ from the SLV is greater than the claimed LOQ of the kit, the method developer must revise the LOQ claimed in the test kit insert and validation reports to meet the

precision requirements for LOQ.

5.2.4.4.3.4. In the validation reports and test kit inserts, the method developers must indicate the actual RSD i value estimated for the LOQ of the kit as part of the

LOQ information. For example:

5.2.4.4.3.4.1. LOQ 15 , for a method where the existing LOQ claimed by the kit had an

estimated RSD i of 15% in the SLV

5.2.4.4.3.4.2. LOQ 30 , for a method where the LOQ was set based on the SLV outcome

and a maximum RSD i of 30%

5.2.5. Robustness Study 608 The Robustness study is intended to provide information on (1) robustness and (2) product stability and 609 consistency. 610 5.2.5.1. Acceptable Test Materials 611 5.2.5.1.1. Spiked matrices are acceptable for test kit lot-to-lot stability analysis and robustness 612 analysis (except when varying extraction conditions). See Annex A for description of 613 best practices for spiked matrix preparation. 614 5.2.5.1.2. Incurred matrices may also be used for the robustness study. If sufficient quantities of 615 incurred matrices have been prepared for the matrix study, these samples may also 616 be used for the robustness studies (i.e., separate incurred matrices are not 617 required). 618