AOAC SPSFAM ERP for SELECTED FOOD ALLERGENS

and concisely? If no, please specify the needed revisions.

by an end user. The authors appear to be suggesting that the amount of allergen in an unknown sample would be determined by comparing the sample area ratio (sample peptide area/heavy internal standard area) to the calibration curve constructed from the incurred/spiked food area ratio. Is that correct? Would the authors intend for each user to run the incurred/spiked food calibration curve? If so, will those materials be available for end users? Also, if that is the intended quantification method to obtain a result, why did the authors use different quantitative comparisons in their recovery analysis? Lastly, if the quantification strategy uses the incurred/spiked foods as the calibration curve, then the quantitative data shown in the submission only supports the performance of the method with the standard curve itself, not on any sort of unknown or reference samples. In addition, the authors initially show the results for just the one quantifier ion for each allergenic food (Tables 10-14), but then go on to show quantitative results for the qualifier ion in Appendix 4. How were those results based on the qualifier ion calculated? Did the authors also have heavy internal standards for the qualifier ion? The method and the supporting information are also not clear on how the data from the other transitions described as being monitored in the method (from the other peptides and/or proteins) would be assessed. Supposedly two transitions for each of two peptides from two proteins from the allergenic source were monitored, but no data for other transitions/peptides/proteins (aside from the qualifier/quantifier ions) are presented in the supporting information. Does confirmation of presence of the allergenic food require detection of all of the transitions or only some of them?