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1. Based on the supporting
information, were there any
additional steps in the
evaluation of the method that
indicated the need for any
additional precautionary
statements in the method?
In my opinion there is no need.
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.
YES: There are.
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.
YES: There are information demonstrating that the method system suitability tests and
control as specified in the SMPR worked appropriately and expected.
4. Based on the supporting
information, is the method
written clearly and concisely?
If no, please specify the
needed revisions.
The method is well described and substantively prepared. The project of the method is
well integrated and includes a clear and concise description.
5. Based on the supporting
information, what are the
pros/strengths of the
method?
The developed method was evaluated following the definitions of AOAC SMPR
2016.002 with respect to Method quantitation limit (MQL), Method detection limit
(MDL), Linearity, Repeatability, Reproducibility, Recovery. Analytical data was
collected for allergen/matrix combinations listed in AOAC SMPR 2016.002.
Specificity is another important analytical parameter. Characteristic signature peptides
were chosen for each allergen, and MRM transitions for each signature peptide were
determined based on their uniqueness compared to background proteins and their
sensitivity of detection. For each allergen, multiple unique peptides were chosen out of
unique proteins, and two MRM transitions per peptide were chosen.
6. Based on the supporting
information, what are the
cons/weaknesses of the
method?
The cons/weakness of the method may be costs. But I think it is inevitable.
7. Any general comments
about the method?
My remarks to the AOAC Candidate Method #ALL-01 are as follows:
Page 13, LC-MS Separation:
The authors should answer the following question:
How long stationary phase was conditioned by mobile phase?
Moreover, they should add details about the condition of the stationary phase:
- before the analysis,
- between each of the analyses in gradient elution mode (if the additional conditioning
was applied for balancing of chromatographic system, despite the fact that
concentrations of both components of mobile phase on the start and the end of
gradient are the same).
I have no additional comments.