AOAC SPIFAN Nutrients ERP Reviewer Forms (December 7, 2022)

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Expert Review Panel for Infant Formula and Adult Nutrition Evaluation of Method HMO -02 (Resubmission)

Title: Determination of Seven Human Milk Oligosaccharides (HMO) in Infant Formula and Adult Nutritionals

Author: T. Bénet, N. Frei, V. Spichtig, D. Cuany, S, Austin Reviewer Name: Reviewer 1

Please Provide a Brief Summary of the Resubmitted Method: The determination of seven human milk oligosaccharides (2’FL, 3FL, 3’SL, 6’SL, DFL, LNT and LNnT) in infant formula and adult nutritionals is described. All oligosaccharides are labeled at their reducing end with 2 aminobenzamide (2AB), separated by means of liquid chromatography and detected by using a fluorescence detector. Consideration of interfering components such as, lactose-3’-phosphate, maltodextrins, galactooligosaccharides (GOS), polydextrose and fructooligosaccharides is taken and solutions for limiting interference are described. The method has been validated in a single laboratory on infant formula (e.g. cow, goat, soy milk based) and adult nutritionals. Recoveries were in the range of 90.9-109% in most cases. Relative repeatabilities (RSD r ) were in the range 0.1-4.2%. Was Method Scope/Applicability Updated? No, the scope and applicability of the method was not changed or updated, as it was not required. The method is applicable on most infant formula and adult nutritionals. However, there are some limitations in regard to 6’SL determination in soy-based formula and LNT/LNnT in elemental formula.

Has the resubmitted method met the requirements outlined as per the ERP recommendation/comment(s)? The authors provided answers and proceeded to corrections in the method as requested/suggested from the ERP and reviewers.

Please provide documentation:

Provide General comments: In general, the method is well written and relatively easy to understand. The authors have updated the method and corrected what was suggested by the ERP and reviewers. Several tables and figures are supporting the text and particularly the validation work. In addition to that, additional supporting material was provided – as it was requested. The necessary method performance requirements as obtained by the single laboratory validation

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