AOAC SPSFAM ERP for SELECTED FOOD ALLERGENS

Submission Date

Name

E-mail

Organization

Title of Method

AOAC Candidate Method

Number (e.g. ALN-01)

Applicable SMPR

I. General Summary:

1. Does the applicability of

the method support the

applicability of the SMPR? If

not, please explain what is

missing.

2. Does the analytical

technique(s) used in the

method meet the SMPR? If

not, please specify how it

differs from what is stated in

the SMPR.

AOAC SPSFAM ERP REVIEW FORM - MARCH 13, 2017

2017-03-01 19:29:16

SNEH BHANDARI

sneh.bhandari@mxns.com

Silliker Laboratories

Detection and Quantitation of Selected Food Allergens using LC-MS/MS (Revision 2)

ALL-01

2016.002

A LC-MS/MS based method for the detection and quantitation of whole egg (egg white

and egg yolk), whole milk, peanut butter, and hazelnut commodity in food matrices

listed in AOAC SMPR 2016.002 was developed and tested. The method uses triple

quadrupole mass spectrometry and Multiple Reaction Monitoring (MRM) of

characteristic transitions of precursor ions to fragment ions of a marker peptides to

uniquely identify each allergen. The calibration curves were plotted using the ratio of

unique peptide peak area of each allergen commodity in food matrix to spiked labeled

internal standard (area ratio) against incurred or spiked commodity concentration. The

recoveries (%) for each allergen commodity were estimated (n=9). The same quantifier

ion was used for the different food matrices except chocolate and wine due to matrix

interference or non-specific binding for certain proteins or peptides. The sensitivity and

selectivity of the method are demonstrated.. Overall, the method was able to meet the

method performance requirements stated in AOAC SMPR 2016.002. For all food

matrices, the target commodity analytical range of 10

‒

1000 ppm was achievable and

the method demonstrated good repeatability with RSDr < 15% . Except for chocolate,

the peptide recoveries ranged from 60.5 to 109.9% in the tested food matrices. The

method is able to demonstrate good sensitivity and is able to detect whole egg and the

rest of the allergen commodities at a MQL of 3 or 10 ppm, respectively. Similar results

were also achieved for the qualifier ion. Except for quantifier ion, poor sensitivity (MQL

≤ 30%) and low recovery (< 60%) were observed for the qualifier ions of milk, peanut

and hazelnut in chocolate due to severe matrix effects. The scope of the method is

limited to these matrices and allergen commodities.

SMPR states detection and quantitation of egg, milk, peanut, and hazelnut food

allergens in finished food products and ingredients. The submitted method is able to

detect mentioned allergens in selected matrices. Egg white and egg yolk in Cookies,

bread, cookie dough, salad dressing and white wine. Whole milk in cookies, infant

formula, red wine and dark chocolate. Peanut and hazelnut in cookies, ice cream,

breakfast cereals and milk chocolate. The method has not been applied to different

varieties of specified matrices in respective allergen category.

Yes.

II. Review of the

Method Only: