APAC SPSFAM Mid Year 2016

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”? 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”?

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. 

11 50

Discuss in the working group maybe follow ELISA guidelines

Reference to Appendix M:  Validation Procedures for  Quantitative Food Allergen ELISA Methods added to SMPR.

12 46

Discuss in the working group maybe follow ELISA guidelines

Line 108 of version  revised  to recommend "LOQ, MDL, recovery  and precision" data for every claimed matrix.

include a sentence for each parameter that explains the parameter‐ specific validation

13 46‐69

LoQ, MDL, recovery and precision data need to be determined for every claimed matrix

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.

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.  

116 (table  1)

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.

Change MDLs/LoQ in table 1 according to the VITAL values and  calculations given under comments. Discuss in the working group

14

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.

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)

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).

No change.   The working group did not agree.

15

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. 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

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

17

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?

No change.   AOAC policy not a working group decision.

18 9

discuss in the working group

Change title to “…selected food allergens."

19 3

The title is unclear

change to “…selected food allergens”

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?

No change.   Method comparision is not a verification  requirement.

20 9

Include minimum requirements for verification