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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.
See previous answer indicating no specific system suitability tests apparent.
4. Based on the supporting
information, is the method
written clearly and concisely?
If no, please specify the
needed revisions.
The method was submitted as two separate documents, which makes it difficult for a
reader to understand the final method workflow proposed by the authors. In addition,
the method is not presented in a way that would be easy for an end user to read and
follow. The authors should submit a more precise, stepwise explanation of the method
from start to finish (e.g. including all final parameters for extraction, digestion, clean-
up/SPE, LC-MS/MS, and data analysis.
The method does not provide sufficient information as to how the authors conducted
the data analysis or how an end user would conduct the data analysis. For example,
which peptides and transitions were used for quantification? How should an end use
choose those? In the transition list table, the authors indicate at least two peptides that
would be used for quantification, but the supporting data only shows results from one
peptide from each allergenic food. How was that selection made, and how would end
users make that selection? Also, one would assume that the authors are calculating the
peak areas for the monitored transitions, but no information is given about the
parameters the authors used or what end users should do to derive the quantitative
information. Did the authors require a minimum number of points across the peak, for
example?
The method also needs to have substantially more information about how calibration
curves were prepared and how end users would prepare calibration curves for the
method. What exact material (e.g. lyophilized whole milk or fluid whole milk) was used
for the calibration curves and what were the curves prepared in (in water or in some
sort of buffer or in matrix-matched extracts)? Was each concentration extracted and
prepared separately or were dilutions performed following digestion and SPE cleanup?
How do the authors plan to address the presence of target peptides in species other
than the target species (hen’s egg peptides in turkey; hazelnut peptide h4 in peach;
and Bos Taurus peptides in water buffalo and wild yak?
5. Based on the supporting
information, what are the
pros/strengths of the
method?
It is possible that this submission could be the beginning of a sound method, but the
lack of clarity on several key issues noted in this review, makes it difficult to actually
assess the method.
6. Based on the supporting
information, what are the
cons/weaknesses of the
method?
The final procedures for the method are unclear, in part due to two separate
documents being submitted at different times and, in part, due to a general lack of
precision and clarity in the writing and presentation of information. While there are a
number of items that need to be clarified, the largest overarching issue with the
presentation of the method is the lack of complete, accurate, and clear quantitative
units and descriptions of materials used throughout presentation.
7. Any general comments
about the method?
Clear and accurate units must be provided not only for reviewers to evaluate the
effectiveness of the method, but also so that the method delivers correct and usable
data to end users. This method requires significant revision on the issue of units as well
as general clarity before it should be considered again.