Allergens ERP Book, Sept 17

2. Does the method contain system suitability tests or controls as specified by the SMPR? If not, please indicate if there is a need for such tests or controls and which ones. 3. Is there information demonstrating that the method system suitability tests and controls as specified in the SMPR worked appropriately and as expected? If no, please specify. 4. Based on the supporting information, is the method written clearly and concisely? If no, please specify the needed revisions. 5. Based on the supporting information, what are the pros/strengths of the method? 6. Based on the supporting information, what are the cons/weaknesses of the method?

Yes suitability tests and control have been used along the work.

----

Yes, clarity has been significantly improved in this version.

---

A limitation is the use of peanut butter in place of roasted peanuts. In the SPMR it is infact recommended the use of peanuts seeds (from raw and roasted peanuts) for method development. The method demonstrated performed quite well for certain matrices but its applicability was not confirmed for other combination of allergen/matrices. The introduction of another confirmation peptide could be important, likely arising from another protein belonging to the same allergenic ingredient. The authors can make some efforts in optimizing the sample prep protocol to improve sensitivity and recovery for other combinations allergens/matrix for which the SMPR were not met (e.g. chocolate, ice cream)

7. Any general comments about the method?

----

V. Final Recommendation Do you recommend this method be adopted as a First Action and published in the Official Methods of Analysis of AOAC INTERNATIONAL? Please specify rationale.

The authors improved significantly the method that in the current version meets the SMPRe for some allergens. However one negative point remains the use of peanut butter instead of peanut seeds and mor ein general the use of just one peptide for allergen identification excluding any additional peptide for allergen confirmation in the same commodity. I would ask the authors to rework the method adding at least another confirmatory peptide for allergen identification and to show the ratio between qualifier and quantifier transition as they stated in the manuscript.

After a few amendments the method could be considered suitable to be adopted as First Action.