AOAC SPSFAM ERP for SELECTED FOOD ALLERGENS

chromatographic gradient (2-40% acetonitrile).

With respect to how the transition data was processed and analyzed, particularly with respect to the quantitative analysis, some aspects require additional information. The authors seem to recommend quantification by developing a specific calibration curve for each allergenic food target and food matrix combination, using incurred/spiked foods. The quantification calibration curve would utilize a ratio of the spiked/incurred peptide peak area to the peak area of an internal heavy standard peptide spiked in following sample digestion. The concentration in an unknown sample would be determined by including the heavy internal standard peptide prior to analysis, and determining the subsequent light(sample):heavy peak area ratio and comparing that to the calibration curve. What remains unclear is how an end user would implement this calibration curve process in order to conduct the method. Will the end user need to analyze the spiked food calibration curve on their own instrument? If so, will the method developers provide the calibration curve materials? Also, how similar must the calibration curve food matrix be to the unknown sample food matrix? The authors have only shown information about the one set of food matrices that they prepared for the calibration curve itself, not for other food products. It remains quite unclear how an end user would conduct the quantitative analysis. In addition, the authors did not provide much information about how each transition and/or peptide would be evaluated either qualitatively or quantitatively in this system. For example, is the sum of the individual peptide transitions utilized for the peak area calculations? Are both transitions for each peptide required? In addition, it is unclear how many peptides and transitions are actually monitored and quantified in the final method. It is initially stated that two transitions are monitored from two peptides