AOAC HMO-06 Reviewer Forms (July 25, 2023)

FOR EXPERT REVIEW PANEL USE ONLY NOT FOR DISTRIBUTION

1. Is the Validation Study Report in a format acceptable to AOAC? Yes

2. Is the method described in sufficient detail so that it is relatively easy to understand, including equations and procedures for calculation of results (are all terms explained)? Yes

3. Are the figures and tables sufficiently explanatory without the need to refer to the text? No – In some cases the titles could be improved and it is not always easy to understand exactly what data is in the tables. 4. Are all the figures and tables pertinent? YES – I would also like to see plots of residuals next to calibration curves (they are easier to read than tables)

5. Could some be omitted and covered by a simple statement?

No

6. Are the references complete and correctly annotated? Yes. However I would like to see an additional reference to support the statement “ The most widely adopted reaction for this task is reductive amination, which has been used to determine HMOs from human milk as well as infant formula” (e.g. Anumula 2006 Anal Biochem 350: 1-23) or some papers analyzing HMOs using such approaches (there are plenty of examples).

7. Does the method contain adequate safety precaution reference and/or statements? Yes

Recommendation: Recommend no action at this time. Method looks promising, however the validation is insufficient. Authors should use wider range of formula matrices and multiple spike levels (preferably covering the range in the SMPR). Also the LoQs should be assessed in-matrix, not in water.

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