AOAC SPIFAN Stakeholder Panel Meeting Book (March 12, 2020)

Summary of Existing Vitamin D Methods – OMA, Chapter 50

Challenges with Existing Methods Methods Scope and Performance vs Current SMPR Are Methods Still in Use? Do any of the fit-for-purpose methods still have utility? Is more than one SMPR needed based on intended use or are any revisions needed for the current SMPRs Any gaps in types of methodology needed based on review Other findings

Summary of Existing Methods in OMA, Chapter 50 Method Status Method Applicability Comments 992.26 “Vitamin D 3

• AOAC Final Action • Codex Type II (Special Foods)

• Vitamin D 3 • Range: 0.25 – 3.75 mcg/100mL • Milk and Soy-Based IF Studied • Applicable to wide range of SPIFAN matrices • Recovery and RSD r pass SMPR • RSD R does not pass SMPR • Vitamin D (D 2 and D 3 ) • Range: 0.02 – 7.3 mcg/100mL • Applicable to wide range of SPIFAN matrices • RSD r and RSD R does not meet SMPR for all matrices studied

• Sap, L/L Extraction, NP-NP LC-UV; External calibration • Pre-D not measured, mathematical accounting via kinetics • SLV: Milk- and Soy-based IF • MLT: Three RTF Milk-Based IF ; RSD R does not meet SMPR • RSD R : Does not meet SMPR for any of three products • SLV / MLT not completed on SPIFAN Kit matrices • Sap, L/L Extraction, SPE Clean up, RP LC-UV Analysis • Internal Standard (D 2 , D 3 ) • Pre-D not measured but accounted for via kinetics • SLV: IF Liq/Pwd, Enteral Liq/Pwd’ MLT: 5 Product Types • SLV / MLT not completed on SPIFAN Kit matrices • Sap, L/L Extraction; NP LC Clean up; RP LC-UV Analysis • Internal Standard (D 2 ) • Pre-D not addressed • SLV/MLT – RSD r and RSD R pass SMPR (One IF) • SLV / MLT not completed on SPIFAN Kit matrices

(Cholecalciferol) in Ready-to-Feed Milk- Based Infant Formulas”

995.05 “Vitamin D in infant Formulas and Enteral Products”

• AOAC First Action • Codex Type III (Infant Formula)

2002.05 “Cholecalciferol (Vitamin D 3 ) in Selected Foods”

• AOAC First Action

• Vitamin D 3 Only • Range: 0.4 – 12 mcg/100g • Horizontal method • One Infant Formula Studied

LC-UV Based methods Common well-characterized equipment and rugged sample procedures Selectivity driven by chromatography – Sample matrix challenges Internal Standardization Used in two methods Still in use Lacks selectivity of newer technology (MS/MS)