AOAC SPIFAN Nutrients ERP Reviewer Forms (July 28, 2022)

FOR EXPERT REVIEW PANEL USE ONLY NOT FOR DISTRIBUTION

Expert Review Panel for Infant Formula and Adult Nutrition Evaluation of Method Phospho-02 July 2022

Title: Quantification of phospholipids by UPLC-MS/MS Author: Francesca Giuffrida and Isabelle Tavazzi, Societe des Produits Nestle S.A. Reviewer Name: Reviewer 2

Summary of Method: Phospholipids are extracted using organic solvents and separated according the PL head groups on a silica column using a gradient mobile phase. Chromatography and detection by LC-MS/MS using electrospray ionization source. Quantification is conducted using external calibration method. All molecular species from each phospholipid class are grouped and quantified together. Method Scope/Applicability: Applicable to the determination of phospholipids in Infant formula and Adult nutrition (phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), and sphingomyelin (SM)) General comments about the method: The method takes advantage of MS technology to selectivity detect and quantify classes of PL’s by their head groups. The method was developed and published in 2018 and has been used since then. Compared with 2018 separation of Phosphatidylserine was improved. The quantification using external calibration method uses selected calibrants expected for infant formulas and adult nutritional’s and is straightforward.

Method Clarity: The method procedure is easy to follow. A system suitability section is included. We have not seen a specific safety section.

Pros/Strengths: • Selective detection of PL classes using LC-MS/MS • High detection sensitivity • Mix of standards designed and applicable for infant formulas and adult nutrition (and ingredients is characterized).

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