AOAC ISPAM Food Allergens Egg/Milk SMPR Info to WG

AOAC INTERNATIONAL INTERNATIONAL STAKEHOLDER PANEL ON ALTERNATIVE METHODS (ISPAM) Food Allergens Standard Method Performance Requirements (SMPR®) & Annexes

September 13, 2017

AOAC INTERNATIONAL 2275 Research Blvd., Suite 300 Rockville, MD, 20850

UNITED STATES dboyd@aoac.org 301.924.7077 x126

AOAC INTERNATIONAL International Stakeholder Panel on Alternative Methods (ISPAM)

Meeting at the Atlanta Marriott Marquis 265 Peachtree Center Ave NE, Atlanta, GA 30303 , USA STAKEHOLDER PANEL DRAFT MEETING AGENDA Sunday, September 24, 2017 Meeting Start Time: 8:30AM (Eastern US) ISPAM Chair: Erin Crowley (Q Laboratories, Inc.)

Location: Salon M106/107

(Registration Opens at 7:30AM)

I. WELCOME & INTRODUCTIONS (Goodwin/Crowley – 8:30AM-8:45AM) Jonathan Goodwin (AOAC) will open the meeting by welcoming attendees, leading introductions, and introducing ISPAM Chair, Erin Crowley (Q Laboratories, Inc.). Crowley will call the meeting to order.

II. AOAC ISPAM GOALS/OVERVIEW/UPDATE (Crowley – 8:45AM-9:15AM)

Erin Crowley will review the AOAC ISPAM meeting agenda and goals. She will also provide an overview of ISPAM and its working group activities, including an update on development of the Standard Method Performance Requirements (SMPR®) for Egg, Milk and Gluten in Oats. She will also provide an overview of ISPAM activities between March 2017 and September 2017.

III. AOAC STANDARDS DEVELOPMENT POLICIES & PROCEDURE OVERVIEW (McKenzie – 9:15AM-9:30AM) Deborah McKenzie (AOAC) will provide information on the AOAC Standards Development process.

IV. MICROBIOLOGICAL METHOD VALIDATION FOR CANNABIS (Wong – 9:30AM-10:30AM) Seth Wong (TEQ Analytical Laboratories) will describe current practices and gaps for Cannabis.

V. AOAC ISPAM WORKING GROUP ON QUANTITATIVE MICROBIOLOGY METHODS ACCEPTANCE CRITERIA (Bird/Arbault – 10:45AM-12:00PM) Patrick Bird (Q Laboratories) & Patrice Arbault (BioAdvantage) Working Group Co-Chairs, will provide information on the development of AOAC Quantitative Microbiology Method Validation Acceptance Criteria. VI. AOAC PRESIDENTIAL TASK FORCE ON FOOD FRAUD (Godefroy – 1:30PM-2:00PM) Samuel Godefroy (Université Laval) will inform AOAC ISPAM on the efforts of an AOAC Task Force relating to Food Fraud.

VII. WORKING GROUP ON FOOD ALLERGENS ASSAYS – MILK (Godefroy/Yeung – 2:00PM-2:30PM) Samuel Godefroy (Université Laval) & Jupiter Yeung (Nestlé) will present the status of the draft SMPR® for Milk.

*Requires a vote Draft meeting agenda is subject to change w/out notice

Version 3.2

VIII. WORKING GROUP CHAIR PRESENTATIONS AND VOTE ON FINAL SMPR® (Working Group Chairs – 2:30PM- 4:30PM) *Each Working Group Chair will present the work of the respective working group and how it reached consensus, and present the SMPR® for vote by the AOAC ISPAM Stakeholders.

1. WORKING GROUP CO-CHAIRS, Samuel Godefroy (Université Laval) & Jupiter Yeung (Nestlé)  FOOD ALLERGENS ASSAY (EGG)

2. WORKING GROUP CHAIR, Joe Boison (CFIA)  GLUTEN IN OATS

IX. NEXT STEPS (Crowley/McIver – 4:30PM-4:45PM) Erin Crowley (Q Laboratories) and Krystyna McIver (AOAC) will discuss next steps for the working group activities, wrap up all discussions and answer any additional questions.

MEETING ITINERARY: Registration Meeting Start Time

(7:00AM) (8:30AM) (10:30AM) (12:00PM) (3:15PM)

Morning Break

Lunch Own Your Own

Afternoon Break

*Requires a vote Draft meeting agenda is subject to change w/out notice

Version 3.2

DRAFT AOAC SMPR® for Egg Allergen Version 8; March 13, 2017 1 2 Quantitation of Chicken Egg by ELISA-based Methods 3 4 Intended Use : Method for quantitation of chicken egg in the context of food manufacturing. 5 6 1. Purpose: AOAC SMPRs describe the minimum recommended performance characteristics to 7 be used during the evaluation of a method. The evaluation may be an on-site verification, a 8 single-laboratory validation, or a multi-site collaborative study. SMPRs are written and 9 adopted by AOAC Stakeholder Panels composed of representatives from the industry, 10 regulatory organizations, contract laboratories, test kit manufacturers, and academic 11 institutions. AOAC SMPRs are used by AOAC Expert Review Panels in their evaluation of 12 validation study data for method being considered for Performance Tested Methods or AOAC 13 Official Methods of Analysis , and can be used as acceptance criteria for verification at user 14 laboratories.

15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46

2. Applicability:

Quantitation of chicken egg in one or more food(s) such as those listed in table 3 of

Appendix M. 1

3. Analytical Technique :

Enzyme-linked immunosorbent assay (ELISA)-based assays (see definition in section 4).

4. Definitions :

Enzyme-linked immunosorbent assay (ELISA)

For the purposes of this document, ELISA is defined as “an analytical procedure characterized by the recognition and binding of specific antigens by antibodies” 1 . This definition is not meant to be restrictive and encompasses other related binding based

technologies.

Limit of Detection (LOD)

LOD is defined as the lowest concentration or mass of analyte in a test sample that can be distinguished from a true blank sample at a specified probability level . 2

Limit of Quantitation (LOQ)

LOQ is the lowest level of analyte in a test sample that can be quantified at a specified level

of precision. 3

Repeatability

Variation arising when all efforts are made to keep conditions constant by using the same instrument and operator (in the same laboratory) and repeating during a short time period.

Expressed as the repeatability standard deviation (SD r

); or % repeatability relative standard

deviation (%RSD r

). 4

47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97

Reproducibility

Variation arising when identical test materials are analyzed in different laboratory by different operators on different instruments. The standard deviation or relative standard deviation calculated from among-laboratory data. Expressed as the reproducibility standard

deviation (SD R

); or % reproducibility relative standard deviation (% RSD R ). 5

Recovery

The fraction or percentage of analyte that is recovered when the test sample is analyzed

using the entire method.

Egg

A combination of [chicken] egg whites and egg yolks in their entirety, in natural proportions. 6 For the purposes of this SMPR, egg is referred to in its dry form as represented

by existing reference materials

5. Method Performance Requirements :

See table 1.

6. System suitability tests and/or analytical quality control:

See antibody information, cross reactivity, information on calibrators and information on matrices in section “Required Allergen-Specific Information to be Provided on the ELISA

Method” of Appendix M.

Method developers should clearly identify what component(s) of the egg they are measuring and provide the conversion factor used to equate to dried egg.

Method developers should provide applicability statement for intended use and claimed

matrices.

7. Reference Material(s):

Refer to Annex F: Development and Use of In-House Reference Materials in Appendix F: Guidelines for Standard Method Performance Requirements , 20 th Edition of the AOAC

INTERNATIONAL Official Methods of Analysis (2016). Available at:

http://www.eoma.aoac.org/app_f.pdf

Chicken Egg

• NIST 8445 (Spray dried whole egg for allergen detection)

8. Validation Guidance :

Method developers must provide data for method performance in all the claimed matrices.

Method developers must provide recovery data using incurred samples for all claimed

matrices.

Appendix D: Guidelines for Collaborative Study Procedures To Validate Characteristics of a Method of Analysis; 20 th Edition of the AOAC INTERNATIONAL Official Methods of Analysis

(2016). Available at: http://www.eoma.aoac.org/app_d.pdf

Appendix F: Guidelines for Standard Method Performance Requirements; 20 th Edition of the AOAC INTERNATIONAL Official Methods of Analysis (2016). Available at:

http://www.eoma.aoac.org/app_f.pdf

98 99

Appendix M: Validation Procedures for Quantitative Food Allergen ELISA Methods: Community Guidance and Best Practices; 20 th Edition of the AOAC INTERNATIONAL Official Methods of Analysis (2016). Available at: http://www.eoma.aoac.org/app_m.pdf

100 101 102 103 104

Table 1: Method performance requirements

Parameter

Minimum Acceptance Criteria for target matrix

Analytical Range (ppm)

≤5 - ≥10

LOQ (ppm)

≤5

LOD (ppm)

≤5

50-150%

Recovery (%)*

% RSD r

≤20 %

% RSD

≤ 30%

R

Note: ppm dried egg. *Using incurred samples (acceptance criteria in Appendix M).

105 106 107 108 109

1. Appendix M: Validation Procedures for Quantitative Food Allergen ELISA Methods: Community Guidance and Best Practices, Official Methods of Analysis (2016) 20 th Ed., AOAC INTERNATIONAL, Rockville, MD, www.eoma.aoac.org; and, Journal of AOAC INTERNATIONAL [ 93 (2) 442-450] (2010). 2. Ibid. 3. Ibid. 4. ISO 5725-1:1994 5 I bid. 6. Introduction to Egg Products, USDA Food Safety and Inspection Service, website:

http://www.fsis.usda. gov/wps/wcm/connect/c5c85914-5055-4f09-8098- 1a179a1c6e14/EPT_Introduction.pdf?MOD=AJPERES, accessed 12/15/2015.

Annex I - Choice of LOD / LOQ (For Egg SMPR)

Limit of Detection (LOD) and Limit of Quantification (LOQ) are selected based on user requirements. The proposed limits in the SMPR constitute minimum requirements for food allergen testing in the targeted matrices as part of a food processing control. Assay users or developers may want to consider assays with a broader performance range, e.g. more sensitive and/or broader range, than the minimum acceptance criteria, as needed by their applications. For example, some users may seek the lower bound of the analytical range to correspond with either a regulatory or a health-driven threshold limit. Only a few jurisdictions such as Japan have set a regulatory limit of 10 ppm protein for all their priority allergens. Other jurisdictions attempt to rely on risk-based thresholds for the various priority allergens. Nonetheless, recent developments in reference doses 1 have been used by food manufacturers and others as part of risk management approaches that are developed by the food industry sector in Australia and New-Zealand 2 . Even with this new information, the food safety risk assessment community has not adopted a validated food allergen reference or benchmark doses, which can be applied consistently by food regulators and food manufacturers in allergen-related health risk assessments and the management of precautionary allergen labeling. The V oluntary I ncidental T race A llergen L abeling (VITAL) 3 initiative of the Allergen Bureau in Australia and New-Zealand developed an open and transparent scientific approach 1 using reference doses for allergen risk characterization, taking into account of clinical food allergen challenge studies. For each priority allergen targeted, a reference dose is defined as the milligram protein level (total protein from an allergenic food) below which only the most sensitive individuals (between 1% and 5% depending on the quality of the data set available) in the allergic population are likely to experience an adverse reaction . For example, in VITAL 2.0 the reference doses are currently set at 0.2 mg protein for peanut, 0.1 mg protein for milk, and 0.03 mg protein for egg 3 These reference doses are used to generate action levels for food allergen control, taking into account the serving size of the food in an eating occasion. Analytical targets may therefore be set at such action levels or lower. For example, the LOQ or action level for egg protein potentially present in a food consumed at a 100 g serving size would be 0.3 ppm (0.03 mg protein in 100 g).

1 Taylor et al (2014) Establishment of reference doses for residues of allergenic foods: report of the vital expert panel, food and chemical toxicology. Food Chem Toxicol 63: 9–17. 2 The Allergen Bureau was established in 2005 and is funded by membership from the Australian and New Zealand food industry 3 http://allergenbureau.net/vital/ accessed on August 25 th , 2017

DRAFT AOAC SMPR® for Milk Allergen; Version 6, July 20, 2017 1 2 Quantitation of Milk by ELISA-based Methods 3 4 Intended Use : Method for quantitation of milk in the context of food manufacturing. 5 6 1. Purpose: AOAC SMPRs describe the minimum recommended performance characteristics to 7 be used during the evaluation of a method. The evaluation may be an on-site verification, a 8 single-laboratory validation, or a multi-site collaborative study. SMPRs are written and 9 adopted by AOAC Stakeholder Panels composed of representatives from the industry, 10 regulatory organizations, contract laboratories, test kit manufacturers, and academic 11 institutions. AOAC SMPRs are used by AOAC Expert Review Panels in their evaluation of 12 validation study data for method being considered for Performance Tested Methods or AOAC 13 Official Methods of Analysis , and can be used as acceptance criteria for verification at user 14 laboratories.

15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45

2. Applicability:

Quantitation of milk in one or more food(s) such as those listed in Table 3 of Appendix M. 1

3. Analytical Technique :

Enzyme-linked immunosorbent assay (ELISA)-based assays.

4. Definitions :

Enzyme-linked immunosorbent assay (ELISA)

For the purposes of this document, ELISA is defined as “an analytical procedure characterized by the recognition and binding of specific antigens by antibodies”. This definition is not meant to be restrictive and encompasses other related binding based

technologies.

Limit of Detection (LOD)

LOD is defined as the lowest concentration or mass of analyte in a test sample that can be distinguished from a true blank sample at a specified probability level . 2

Limit of Quantitation (LOQ)

LOQ is the lowest level of analyte in a test sample that can be quantified at a specified level

of precision. 3

Repeatability

Variation arising when all efforts are made to keep conditions constant by using the same instrument and operator (in the same laboratory) and repeating during a short time period.

Expressed as the repeatability standard deviation (SD r

); or % repeatability relative standard

deviation (%RSD r

). 4

46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83

Reproducibility

Variation arising when identical test materials are analyzed in different laboratories by different operators on different instruments. The standard deviation or relative standard deviation calculated from among-laboratory data. Expressed as the reproducibility standard

deviation (SD R

); or % reproducibility relative standard deviation (% RSD R ). 5

Recovery

The fraction or percentage of analyte that is recovered when the test sample is analyzed

using the entire method.

Milk

For the purposes of this SMPR: “milk” refers to nonfat dry milk from pasteurized skim milk. It contains not more than 5 percent by weight of moisture, and not more than 1.5 percent

by weight of milkfat, 1 as represented by existing reference materials.

5. Method Performance Requirements :

See Table 1.

6. System Suitability Tests and/or Analytical Quality Control:

See antibody information, cross reactivity, information on calibrators and information on matrices in section “Required Allergen-Specific Information to be Provided on the ELISA

Method” of Appendix M.

Method developers should clearly identify what component(s) of milk they are measuring and specifically the method performance vis a vis the milk component such as whey, beta-

lactoglobulin, or casein.

Method developers should clearly identify what component(s) of milk they are measuring and specifically the method performance with regards to the milk component such as whey, beta-lactoglobulin, or casein; and the target used to generate antibodies.

Method developers should provide the conversion factor used to equate to non-fat dried

milk.

Method developers should provide applicability statement for intended use and claimed

matrices.

1 Code of Federal Regulations; Title 21 - Food and Drugs, § 131.125. Other internationally recognized definition may be applied.

2 MoniQA Association, http://www.moniqa.org/node/910

84 85 86 87 88 89 90 91 92 93 94 95 96 97 98

7. Reference Material(s):

Nonfat milk powder

• MoniQA Association Reference Material SMP-MQA 092014 2

Refer to Annex F: Development and Use of In-House Reference Materials in Appendix F: Guidelines for Standard Method Performance Requirements , 20 th Edition of the Official Methods of Analysis of AOAC INTERNATIONAL (2016). Available at:

http://www.eoma.aoac.org/app_f.pdf

8. Validation Guidance :

Method developers must provide data for method performance in all the claimed matrices. Must specify if samples used for validation are: incurred or spiked; processed (baked) or

99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116

unprocessed (raw).

Method developers must provide recovery data using incurred samples for all claimed

matrices.

Appendix D: Guidelines for Collaborative Study Procedures To Validate Characteristics of a Method of Analysis; 20 th Edition of the Official Methods of Analysis of AOAC INTERNATIONAL

(2016). Available at: http://www.eoma.aoac.org/app_d.pdf

Appendix F: Guidelines for Standard Method Performance Requirements; 20th Edition of the Official Methods of Analysis of AOAC INTERNATIONAL (2016). Available at:

http://www.eoma.aoac.org/app_f.pdf

Appendix M: Validation Procedures for Quantitative Food Allergen ELISA Methods: Community Guidance and Best Practices; 20th Edition of the Official Methods of Analysis of AOAC INTERNATIONAL (2016). Available at: http://www.eoma.aoac.org/app_m.pdf

117 118

Table 1: Method performance requirements

Parameter

Minimum Acceptance Criteria for target matrix

Analytical Range (ppm) 1

≤10 - ≥ 20 2

LOQ (ppm) 1

≤10

LOD (ppm) 1

≤10

50-150%

Recovery (%) 3

% RSD r

≤20 %

% RSD

≤30%

R

Notes: 1 ppm in non-fat dried milk. 2 See Annex 1 for rationale for setting lower limit of range. 3 Use incurred samples as per Appendix M. Incurred materials can be obtained from MoniQA Association.

119 120 121 122 123

1. Appendix M: 20th Edition of the Official Methods of Analysis of AOAC INTERNATIONAL (2016); and Validation procedures for quantitative food allergen ELISA methods: Community guidance and best practices . Journal of AOAC INTERNATIONAL [93(2) 442-450] (2010)

2. Ibid. 3. Ibid. 4. ISO 5725-1:1994 5 I bid.

Annex I - Choice of LOD / LOQ (For Milk SMPR)

Limit of Detection (LOD) and Limit of Quantification (LOQ) are selected based on user requirements. The proposed limits in the SMPR constitute minimum requirements for food allergen testing in the targeted matrices as part of a food processing control. Assay users or developers may want to consider assays with a broader performance range, e.g. more sensitive and/or broader range, than the minimum acceptance criteria, as needed by their applications. For example, some users may seek the lower bound of the analytical range to correspond with either a regulatory or a health-driven threshold limit. Only a few jurisdictions such as Japan have set a regulatory limit of 10 ppm protein for all their priority allergens. Other jurisdictions attempt to rely on risk-based thresholds for the various priority allergens. Nonetheless, recent developments in reference doses 1 have been used by food manufacturers and others as part of risk management approaches that are developed by the food industry sector in Australia and New-Zealand 2 . Even with this new information, the food safety risk assessment community has not adopted a validated food allergen reference or benchmark doses, which can be applied consistently by food regulators and food manufacturers in allergen-related health risk assessments and the management of precautionary allergen labeling. The V oluntary I ncidental T race A llergen L abeling (VITAL) 3 initiative of the Allergen Bureau in Australia and New-Zealand developed an open and transparent scientific approach 1 using reference doses for allergen risk characterization, taking into account of clinical food allergen challenge studies. For each priority allergen targeted, a reference dose is defined as the milligram protein level (total protein from an allergenic food) below which only the most sensitive individuals (between 1% and 5% depending on the quality of the data set available) in the allergic population are likely to experience an adverse reaction . For example, in VITAL 2.0 the reference doses are currently set at 0.2 mg protein for peanut, 0.1 mg protein for milk, and 0.03 mg protein for egg 3 These reference doses are used to generate action levels for food allergen control, taking into account the serving size of the food in an eating occasion. Analytical targets may therefore be set at such action levels or lower. For example, LOQ or action level for milk protein potentially present in a food consumed at a 100 g serving size would be 1 ppm milk protein (0.1 mg protein in 100 g).

1 Taylor et al (2014) Establishment of reference doses for residues of allergenic foods: report of the vital expert panel, food and chemical toxicology. Food Chem Toxicol 63: 9–17. 2 The Allergen Bureau was established in 2005 and is funded by membership from the Australian and New Zealand food industry 3 http://allergenbureau.net/vital/ accessed on August 25 th , 2017