AOAC SPIFAN Nutrients ERP (December 7, 2022)

a) Analysis of CRM or Reference Material 1 • Where a CRM is not available, a suitable reference material may be substituted and compared to the 2 established levels (see paragraph 3c). 3 • A minimum of nine (9) independent replicates of CRM or reference material should be tested across 4 three (3) days (e.g. triplicate over three days) and compared to certified or reference values. 5 b) Spike recovery 6 • Recovery will be determined from a range of infant formula and adult nutritional matrices (use of the 7 recognized SPIFAN kit is recommended where appropriate, otherwise the Study Directors may agree 8 on the sample types to be used for recovery studies). 9 • Each selected matrix will be spiked at three levels; low, middle and high end of calibration range. Use 10 spike levels covering the analytical range specified in the SMPR. 11 • Spiked and unspiked samples will be analyzed in triplicates on each of three (3) days. 12 • The daily mean of unspiked samples will be used as the innate amount for calculating individual daily 13 recoveries. 18 2. ISO 13528: Statistical methods for use in proficiency testing by interlaboratory comparison. Annex 19 B 20 3. NACRW Reference Material Use in Trace Analysis, NACRW Reference Materials Working Group, 21 Edition 1; 1-20-2021 22 4. ISO17034:2016: General requirements for the competence and consistent operation of reference 23 material producers 24 5. ISO-5725-1:1994: Accuracy (trueness and precision) of measurement methods and results – Part 25 1: General principles and definitions 26 6. ISO 5725-3: Accuracy (trueness and precision) of measurement methods and results – Part 3: 27 Intermediate measures of the precision of a standard measurement method 28 7. V. Barwick (Ed), Eurachem/CITAC Guide: Guide to Quality in Analytical Chemistry: An Aid to 29 Accreditation (3rd ed. 2016). Appendix B-Instrument calibration and performance checks. ISBN 30 978-0-948926-32-7. Available from www.eurachem.org 31 8. Youden, W.J., & Steiner, E.H. (1975) Statistical Manual of the AOAC , AOAC INTERNATIONAL, 32 Gaithersburg, MD 33 9. Vander Heyden, Y., Nijhuis, A., Smeyers-Verbeke, J., Vandeginste, B.G.M., & Massart, D.L. 34 (2001). Guidance for robustness/ruggedness tests in method validation. Journal of 35 pharmaceutical and biomedical analysis . 24, 723 – 753 36 10. Dong, F. (1993). On the identification of active contrasts in unreplicated fractional factorials. 37 Statistica Sinica 3, 209 – 217 38 11. Magnusson and U. Örnemark (eds.) Eurachem Guide: The Fitness for Purpose of Analytical 39 Methods – A Laboratory Guide to Method Validation and Related Topics, (2nd ed. 2014). ISBN 40 978-91-87461-59-0. Available from www.eurachem.org 41 12. 11843-2:2000, Capability of detection – Part 2: Methodology in the linear calibration case 42 13. ISO 11843-5:2008, Capability of detection – Part 5: Methodology in the linear and non-linear 43 14 15 9 References 16 17 1. ISO Guide 30:2015, Reference materials – selected terms and definitions. International Organization for Standardization, Geneva, Switzerland (2015)

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