AOAC Gluten Quantitative Validation Guidance-Round 1(Nov 2023)

Guidelines for Validation of Quantitative Gluten Methods, with Specific Examples for ELISA Assays

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Contents

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1. 2. 3. 4.

Scope

Applicability

Terms and Definitions

Method Developer Validation Study

4.1. Scope

4.2. Calibration Fit Study 4.3. Selectivity Study

4.4. Matrix Study

10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27

4.5. Data Analysis and Reporting for Matrix Studies 4.6. Acceptance Criteria for Matrix Studies 4.7. Robustness Study; Product Consistency and Stability 4.8. Method Instructions and Required Method Information

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Independent Laboratory Study

5.1. Scope

5.2. Matrix Study Collaborative Study

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6.1. Scope

6.2. Number of Collaborators

6.3. Matrix Study 6.4. Test Materials

6.5. Data Analysis and Reporting 6.6. Acceptance Criteria 6.7. Collaborator Comments

7.

Matrix Extension

7.1. Matrix Extension for Single Lab Validation Studies 7.2. Matrix Extension for Multi-Site Collaborative Studies

Figures 28 Figure 1: Design 1a. Lot: test kit lot, TP: test portion, E: ELISA measurement. Design 1a can be used to estimate intermediate 29 precision and repeatability. 30 Figure 2: Design 1b . Lot: test kit lot, TP: test portion, E: ELISA measurement. Design 1b can be used to estimate intermediate 31 precision, repeatability, and lot-to-lot product consistency. 32 Figure 3: Design 2a. Lot: test kit lot, TP: test portion, E: ELISA measurement. Design 2a can be used to estimate intermediate 33 precision, repeatability, and ELISA variance. 34 Figure 4: Design 2b. Lot: test kit lot, TP: test portion, E: ELISA measurement. Design 2b can be used to estimate intermediate 35 precision, repeatability, ELISA variance, and lot-to-lot product consistency. 36

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Figure 5 : Repeatability Only Design

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Annex A: Selectivity Study Matrices

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Annex B: Preparation of Spiked and Incurred Test Materials

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Annex C: Matrix Categories

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Annex D: Statistical Evaluation