AOAC ISPAM Stakeholder Panel Meeting Book 9-24-17

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); or % repeatability relative standard

Expressed as the repeatability standard deviation (SD r

deviation (%RSD r ).

Reproducibility 2

Variation arising when identical test materials are analyzed in different laboratory by different operators on different instruments. The standard deviation or relative standard deviation calculated from among-laboratory data. Expressed as the reproducibility standard

deviation (SD R

); or % reproducibility relative standard deviation (% RSD R ).

Recovery

The fraction or percentage of analyte that is recovered when the test sample is analyzed

using the entire method.

5. Method Performance Requirements :

See Table 1.

6. System Suitability:

See antibody information, cross reactivity, and information on calibrators in Appendix M .

7. Reference Material(s):

7.1 Samples of oat flour spiked with wheat, rye, and barley for validation studies are available from Paul Wehling at General Mills ( Paul.Wehling@genmills.com ) until a  A sample series shall consist of: 1 sample of unspiked oat flour; 2 samples spiked with wheat; 2 samples spiked with rye; and 2 samples spiked with barley.  Gluten content for wheat, rye, and barley used for spiking will be estimated as suitable neutral source is established.

follows:

Wheat: Gluten=(Dumas nitrogen content) x 5.68 x 0.80 Rye: Gluten=(Dumas nitrogen content) x 5.68 x 0.55 Barley: Gluten=(Dumas nitrogen content) x 5.68 x 0.55

See Estimating Recovery of a Gluten ELISA Kit Method for details on procedures for spiking the flour samples (link to paper). Sample providers shall develop an SOP for

producing, storing and shipping the materials.

7.2 Refer to Annex F: Development and Use of In-House Reference Materials in Appendix F : Guidelines for Standard Method Performance Requirements , 20 th Edition of the AOAC

INTERNATIONAL Official Methods of Analysis (2016). Available at:

http://www.eoma.aoac.org/app_f.pdf

8. Validation Guidance :

For all candidate methods, developers must:

• provide antibody information, cross reactivity data, and information on calibrators

according to Appendix M.

• wherever possible, identify peptide sequences or target epitopes for all antibodies

used.

2 ISO 5725-1