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1. Are the definitions
specified in the SMPR used
and applied appropriately in
the supporting
documentation (manuscripts,
method studies, etc...)? If not,
please explain the differences
and if the method is impacted
by the difference.
YES: The definitions specified in the SMPR were used and applied appropriately in the
supporting documentation (manuscripts, method studies, etc.).
2. Is there information
demonstrating that the
method meets the SMPR
Method Performance
Requirements using the
Reference Materials stated in
the SMPR? If not, then
specify what is missing and
how this impacts
demonstration of
performance of the method.
YES: There is a piece of information demonstrating that the method meets the SMPR
Method Performance Requirements using the Reference Material stated in the SMPR.
3. Is there information
demonstrating that the
method performs within the
SMPR Method Performance
REquirements table
specifications for all analytes
in the SMPR applicability
statement? If not, please
specify what is missing and
whether or not the method's
applicability should be
modified.
YES: There is a piece of information demonstrating that the method performs within the
SMPR Method Performance Requirements table specifications for all analyses in the
SPMR applicability statement.
In the method proposed by the authors, the specification of the following data
according to the SMPR Method Performance Requirements table specifications is as
following:
Calibrations were performed over a range of 1-1000 ppm for each allergen. The
calibration linearity was greater than 0.9978 with calibration residuals within 13% of the
actual value over the calibration range.
Table 1 presents the MQL and MDL for the selected allergen marker peptides. The
MQL met the Method Performance Requirements and equalled 1 on the basis of the
calibration residual results. Going to 0.3 ppm resulted in calibration residuals above
20% difference from the actual level. The MDL’s ranged from 0.1 to 0.05 based on a
signal to noise of 3:1 extrapolated from the 1 ppm calibration level.
Spike recoveries for egg, hazelnut, milk and peanut in the allergen matrix combinations
listed in the AOAC SMPR 2016.002 method performance document was performed a 5
ppm and 50 ppm.
The Tables 2 and 3 list the recovery results for each specified allergen matrix
combination. The 5 ppm spike recovery data ranged from 75-101% with a standard
deviation of 15% or less. The 50 ppm spike recovery data ranged from 90-104% with a
standard deviation of less than 12%.
The repeatability of the method was tested on seven separate extractions-digestions
and LC/MS/MS analysis of the food allergens in a cookie at 5 ppm. Table 5 presents
the results for one peptide for each allergen. The repeatability ranged from 4.5-9.4
percent standard deviation for the four allergens.
I provided my additional remarks in the part ‘7. Any general comments about the
method?’
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 to implement any additional steps in the method
evaluated.