AOAC

Item

Line

Numbers (If

Applicable)

Comment

Proposed Change(s)

Response

139 (table

Chocolate is an important matrix for peanut, hazelnut and milk.

Chocolate should be included into the list of priority allergens. If

chocolate is a known problem than the applicability should clearly

state that chocolate is not possible to measure using the validated

method.

No change.

Chocolate is an optional matrix to be tested for

candidate method that claim to work in chocolate.

2 56‐65

Should the precision data obtained over the whole analytical range? Number of levels?

Describe the validation of precision in a more precise way e.g. include

number of levels and replicates

Additional reference

to Appendix D and F are added

116 (table

By definition the analytical range can only start with an LoQ. MDL only gives a yes or no.

No change recommended.

The comment is true but there is

not any prohibition against the LOQ = MDL.

After validation, LC‐MS/MS methods will be used for comparison with ELISA results. An commercial

ELISA is (often) calibrated to the whole allergenic food while LC‐MS/MS is calibrated to peptides. Is

comparability established via reference materials? (again: traceability of LC‐MS/MS to these RMs is

mandatory!)

Discuss traceability and comparability to ELISA results (note: this SMPR

discuss a possible reference method for cGMP compliance!)

No change.

The working group did not agree to tie LC‐MS/MS

results to ELISA results.

5 96

NIST SRM 2387 is not pure peanut but a mixture of roasted peanut, sugar, partially hydrogenated

vegetables oils and salt. See NIST certificate: protein content is given but not peanut content.

Discuss suitability of this SRM in the working group and give conversion

factor

No change.

That’s will be left up to the methode developer.

6 92

NIST SRM 1549 is superseded by NIST 1549a

Delete NIST SRM 1549

Agree. Replace NIST SRM 1549 wuth 1549a.

7 85

NIST 8445 is a whole egg powder with a given protein content. How should a method developer

trace it to whole egg without conversion factor?

Discuss traceability in the working group and discuss a conversion

factor

Working Group agreed that all results to be "reported as ppm

of the target allergen in food commodity".

8 67

Recovery: What kind of samples is required? Spiked or incurred? For ELISA incurred is preferred.

We should follow the guideline for ELISA which prefer incurred

Add a reference to Appendix M: Validation Procedures for

Quantitative Food Allergen ELISA Methods.

Appendix M does

mandate the use of incurred samples. AOAC policy allows for

both kinds of samples. Method developer discretion.

9 67

Recovery: How should a method developer determine this parameter? By spiking with reference

materials or peptides or a different material. One should remember that it is not allowed to use a

reference material for calibration AND spiking! If peptides are used for calibration, how was

traceability established?

Discuss in the working group and remember to solve the traceability

problem

No change recommended.

Method development issue not

SMPR issue.

10 62

Since reproducibility determination is only possible by a collaborative study, an intra‐laboratory

reproducibility should be defined to ease single‐lab validations at the beginning

Inlcude a new clause after repeatability and describe the validation to

be done

No change.

All previous SMPRs used RSDR and RSDr.

AOAC SPSFAM ALLERGENS DRAFT SMPR ‐ COMMENTS on ALLERGENS SMPR FINAL