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11 50
MDL: How should a method developer estimate this parameter? By using blank matrices or blank
matrices spiked with reference materials/peptides? How many replicates? We have very clear
guidelines for allergen determination by ELISA‐why not for “Reference methods for cGMP
compliance”?
Discuss in the working group maybe follow ELISA guidelines
Reference to Appendix M: Validation Procedures for
Quantitative Food Allergen ELISA Methods added to SMPR.
SMPR will also refer to FDA and/or EPA definition for MDL.
12 46
LoQ: How should a method developer determine or even estimate this parameter? By using
reference materials or peptide solutions or blank matrices or blank matrices spiked with reference
materials/peptides? How many replicates? We have very clear guidelines for allergen
determination by ELISA‐why not for “Reference methods for cGMP compliance”?
Discuss in the working group maybe follow ELISA guidelines
Reference to Appendix M: Validation Procedures for
Quantitative Food Allergen ELISA Methods added to SMPR.
13 46‐69
LoQ, MDL, recovery and precision data need to be determined for every claimed matrix
include a sentence for each parameter that explains the parameter‐
specific validation
Line 108 of version revised
to recommend "LOQ, MDL, recovery
and precision" data for every claimed matrix.
14
116 (table
1)
By taking the latest published VITAL reference doses C18(Food Chem. Toxicol. 63: 9‐17, 2014) it is
obvious that the MDLs/LoQs in table 1 are not sufficient when a food is analyzed that is consumed
in a service size of more than 50 g. Lower MDL/LoQ appropriately to the following table. Note: C19
Hazelnut: Reference dose as protein: 0.1 mg; Reference dose as allergenic food: 0.64 mg;
Minimum concentration to be quantified when consuming 50 g food: 12.8 mg/kg and for 200 g 3.2
mg/kg.
Milk: Reference dose as protein: 0.1 mg; Reference dose as allergenic food: 3.03 mg; Minimum
concentration to be quantified when consuming 50 g food: 60.6 mg/kg and for 200 g 15.2 mg/kg.
Peanut: Reference dose as protein: 0.2 mg; Reference dose as allergenic food: 0.8 mg; Minimum
concentration to be quantified when consuming 50 g food: 16 mg/kg and for 200 g 4 mg/kg.
Whole egg: Reference dose as protein: 0.03 mg; Reference dose as allergenic food: 0.25 mg;
Minimum concentration to be quantified when consuming 50 g food: 4.8 mg/kg and for 200 g 1.2
mg/kg.
Change MDLs/LoQ in table 1 according to the VITAL values and
calculations given under comments. Discuss in the working group
No change.
Working Group discussed on 3/3/2016. There are
multiple VITALs with different maximum permissiable
concentrations. The Working Group consensus is that none of
the VITALs are international concensusn standards, and declined
to reset the LOQs or MDLs based on VITALmaximum
permissiable concentrations.
15
How should a method developer prove that the selected peptides are not “too” specific e.g. a
sequence is used that is not present in every commercially available peanut or hazelnut variety. On
the opposite, if the selected peptides are not specific enough, near botanical relatives are detected
which are maybe not allergenic or regulated (see prunus mahaleb example).
At minimum a chapter describing the known specificities/selectivities
should be provided. (Note: Unknown occurrence of peptides that are
not from a allergenic source will always occur in the future, see also
prunus mahaleb)
No change.
The working group did not agree.
16
What are the minimum performance criteria for peptide selection?
Include criteria for peptide selection or give reference
No change.
The working group did not agree.
17
VITAL values are based on amount of protein per service size. Therefore, the definition of the food
allergens as “food commodities” without mentioning the protein content will establish a non‐
comparability between results obtained by an LC‐MS/MS method and VITAL values.
Include some guidance for the user or let the method developer
describe his way of establishing traceability to VITAL values
No change.
Working Group discussed on 3/3/2016. There are
multiple VITALs with different maximum permissiable
concentrations. The Working Group consensus is that none of
the VITALs are international concensusn standards, and declined
to reset the LOQs or MDLs based on VITALmaximum
permissiable concentrations. AND E25
18 9
collaborative test: It should be critically checked if Appendix D is sufficient in the case where LESS
than 8 participants (and/or LC‐MS/MS machines) are available. Is this still collaborative or
forbidden at all?
discuss in the working group
No change.
AOAC policy not a working group decision.
19 3
The title is unclear
change to “…selected food allergens”
Change title to “…selected food allergens."
20 9
This means a method comparison between the original method (checked by an ERP) and this
method transferred to another lab. Are there any guidelines for this case? What is the minimum
required number of measurements to be sure that both methods are comparable?
Include minimum requirements for verification
No change.
Method comparision is not a verification
requirement.