AOAC ERP Gluten Assays - March 2019
AOAC EXPERT REVIEW PANEL FOR GLUTEN ASSAYS
TUESDAY, MARCH 12, 2019 GAITHERSBURG, MARYLAND USA
AOAC INTERNATIONAL OFFICIAL METHODS OF ANALYSIS SM (OMA) PROGRAM
The Official Methods of Analysis SM (OMA) program is AOAC INTERNATIONAL's premier methods program. The program evaluates chemistry, microbiology, and molecular biology methods. It also evaluates traditional benchtop methods, instrumental methods, and proprietary, commercial, and/or alternative methods. In 2011, AOAC augmented the Official Methods SM program by including an approach to First Action Official Methods SM status that relies on gathering the experts to develop voluntary consensus standards, followed by collective expert judgment of methods using the adopted standards. All methods in the AOAC Official Methods SM program are now reviewed by Expert Review Panels for First Action AOAC Official Methods of Analysis SM status, continuance, repeal, and/or to recommend for AOAC Final Action Official Methods status. The OMA program has undergone a series of transitions in support of AOAC's collaborations, evolving technology, and evolving technical requirements. Methods approved in this program have undergone rigorous scientific and systematic scrutiny such that analytical results by methods in the Official Methods of Analysis of AOAC INTERNATIONAL are deemed to be highly credible and defensible. The methods are published in the Official Methods of Analysis of AOAC INTERNATIONAL and supporting manuscripts are published in the Journal of AOAC INTERNATIONAL . AOAC Official Methods SM program allows for submissions for all proprietary, single and sole source methods. Methods submitted through the PTM-OMA harmonized process also will be reviewed through the O fficial Methods of Analysis SM (OMA) program. Other complementary products and services include expanded consulting services for validation protocol development and AOAC INTERNATIONAL Organizational Affiliate Membership.
AOAC INTERNATIONAL 2275 Research Blvd, Suite 300 Rockville, Maryland 20850 Phone: (301) 924-7077
EXPERT REVIEW PANEL (ERP) GLUTEN ASSAYS
TUESDAY, MARCH 12, 2019 3:00 PM – 5:00 PM MEETING ROOM: SALON F/G GAITHERSBURG MARRIOTT WASHINGTONIAN CENTER GAITHERSBURG, MARYLAND 20878 USA
EXPERT REVIEW PANEL CHAIR: TERRY KOERNER, HEALTH CANADA
I.
WELCOME AND INTRODUCTIONS Expert Review Panel Co-Chairs
II. REVIEW OF AOAC VOLUNTEER POLICIES & EXPERT REVIEW PANEL PROCESS OVERVIEW AND GUIDELINES Deborah McKenzie, Senior Director, Standards Development and Method Approval Processes, AOAC INTERNATIONAL and AOAC Research Institute REVIEW OF METHODS & TECHNICAL RECOMMENDATION FOR OMA 2012.01 The Expert Review Panel will discuss and review the title change for OMA 2012.01. The method authors will be present to present, after which the ERP will discuss the method and render a decision on the possible title change. 1) AOAC OFFICIAL METHOD 2012.01 GLIADIN AS A MEASURE OF GLUTEN IN RICE- AND CORN-BASED FOODS [First Action 2012, Final Action 2016] Original Study Director: R-biopharm
III.
IV. DISCUSS FINAL ACTION REQUIREMENTS FOR FIRST ACTION OFFICIAL METHODS (IF APPLICABLE )
ERP will discuss, review and track First Action methods for 2 years after adoption, review any additional information (i.e., additional collaborative study data, proficiency testing, and other feedback) and make recommendations to the Official Methods Board regarding Final Action status.
V.
DISCUSS UPCOMING MEETINGS AND VOLUNTEER ROLES
VI.
ADJOURNMENT
Page 1 of 1 *Agenda is subject to change. V1
AOAC INTERNATIONAL ● 2275 RESEARCH BLVD, SUITE 300 ● ROCKVILLE, MARYLAND 20850 USA
Official Methods of Analysis SM (OMA) Expert Review Panel MEETING AND METHOD REVIEW GUIDANCE
The AOAC Research Institute administers AOAC INTERNATIONAL's premier methods program, the AOAC Official Methods of Analysis SM (OMA). The program evaluates chemistry, microbiology, and molecular biology methods. It also evaluates traditional benchtop methods, instrumental methods, and proprietary, commercial, and/or alternative methods and relies on gathering the experts to develop voluntary consensus standards, followed by collective expert judgment of methods using the adopted standards. The Official Methods of Analysis of AOAC INTERNATIONAL is deemed to be highly credible and defensible. All Expert Review Panel (ERP) members are vetted by the AOAC Official Methods Board (OMB) and serve at the pleasure of the President of AOAC INTERNATIONAL. In accordance to the AOAC Expert Review Panel Member and Chair Volunteer Role Description all Expert Review Panel members are expected to 1) serve with the highest integrity, 2) perform duties and method reviews, and 3) adhere to review timelines and deadlines.
To assist the ERP Chair and its members, please note the following in preparation for Expert Review Panel meetings and method reviews.
Pre-Meeting Requirements 1. Confirm availability and plan to be present to ensure a quorum of the ERP.
(Please refer to page 25, Quorum Guidelines, Expert Review Panel Information Packet ) 2. Ensure that your laptop, CPU or mobile device can access online web documentation. 3. Be prepared for the meeting by reviewing all relevant meeting materials and method documentation.
In-Person Meeting and Teleconference Conduct 1. Arrive on time.
2. Advise the Chair and ERP members of any potential Conflicts of Interest at the beginning of the meeting. 3. Participation is required from all members of the ERP. All members have been deemed experts in the specific subject matter areas. 4. The ERP Chair will moderate the meeting to ensure that decisions can be made in a timely manner. 5. Follow Robert’s Rules of Order for Motions. 6. Speak loud, clear, and concise so that all members may hear and understand your point of view. 7. Due to the openness of our meetings, it is imperative that all members communicate in a respectful manner and tone. 8. Refrain from disruptive behavior. Always allow one member to speak at a time. Please do not interrupt. 9. Please note that all methods reviewed and decisions made during the Expert Review Panel process are considered confidential and should not be discussed unless during an Expert Review Panel meeting to ensure transparency. Reviewing Methods Prior to the Expert Review Panel meeting, ERP members are required to conduct method reviews. All methods are reviewed under the following criteria, technical evaluation, general comments, editorial criteria, and recommendation status. These methods are being reviewed against their collaborative study protocols as provided in the supplemental documentation. Note: The method author(s) will be present during the Expert Review Panel session to answer any questions.
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Version 1 – OMA ERP Meeting Conduct
Official Methods of Analysis SM (OMA) Expert Review Panel MEETING AND METHOD REVIEW GUIDANCE
Reviewing Methods (Cont’d)
Reviewers shall conduct in-depth review of method and any supporting information. In-depth reviews are completed electronically via the method review form. The method review form must be completed and submitted by the deadline date as provided. All reviews will be discussed during the Expert Review Panel meeting. Any ERP member can make the motion to adopt or not to adopt the method. If the method is adopted for AOAC First Action status, Expert Review Panel members must track and present feedback on assigned First Action Official Methods . Recommend additional feedback or information for Final Action consideratio n. Here are some questions to consider during your review based on your scientific judgment: 1. Does the method sufficiently follow the collaborative study protocol? 2. Is the method scientifically sound and can be followed? 3. What are the strengths and weaknesses of the method? 4. How do the weaknesses weigh in your recommendation for the method? 5. Will the method serve the community that will use the method? 6. What additional information may be needed to further support the method? 7. Can this method be considered for AOAC First Action OMA status? Reaching Consensus during Expert Review Panel Meeting 1. Make your Motion. 2. Allow another member to Second the Motion. 3. The Chair will state the motion and offer the ERP an option to discuss the motion. 4. The Chair will call a vote once deliberations are complete. 5. Methods must be adopted by unanimous decision of ERP on first ballot, if not unanimous, negative votes must delineate scientific reasons. Negative voter(s) can be overridden by 2/3 of voting ERP members after due consideration. 6. All other motions will require 2/3 majority for vote to carry.
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Version 1 – OMA ERP Meeting Conduct
9/13/2018
AOAC Expert Review Panels An Orientation
Deborah McKenzie רב Sr. Dir., Standards Development AOAC INTERNATIONAL Staff Liaison ‐ Official Methods Board
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As a
,
AOAC INTERNATIONAL advances and ,
members, organizations, and experts dedicated to developing and validating and of
by
Analytical Excellence
AOAC Strategic Goals
Core Programs
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AOAC STRATEGIC PLAN
Accessible at AOAC homepage www.aoac.org
Analytical Excellence addresses emerging issues and influence standards development as a global leader in analytical excellence
Standards Development
Official Methods of Analysis SM (OMA) & Performance Tested Methods SM (PTM)
Laboratory Proficiency Testing & Quality Management
Analytical Excellence
Journal of AOAC INTERNATIONAL, OMA, ALACC Guide
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AOAC Method Approval Programs
Official Methods of Analysis SM (OMA) • AOAC’s premiere methods program • Approved methods – published in the Official Methods of Analysis of AOAC INTERNATIONAL (print and online) – Manuscripts published in the Journal of AOAC INTERNATIONAL – First Action and Final Action status
Performance Tested Methods SM (PTM) • AOAC’s method certification program • Certified methods – Commercial/proprietary rapid methods (test kits) – Certifications published on AOAC website – Manuscripts published in the Journal of AOAC INTERNATIONAL – Method developers licensed to use certification mark – Annual review & recertification
AOAC Official Methods SM Program
Submit Methods Responding to issued Call for Methods • Adoption of methods as Official Methods is contingent upon standards development activities • No application fee required to submit methods in response to Call for Methods Submit Individual & Sole Source Methods • Adoption of methods as Official Methods is contingent upon data supporting applicability and community based validation guidance information • Including proprietary/commercial methods and harmonized PTM – OMA methods • Application fee required
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Status of Official Methods of Analysis First Action, Final Action, Repeal
AOAC Policies & Procedures
Policy on Use of Association Name, Identifying Insignia, Letterhead, Business Cards
Policy on Volunteer Conflict of Interest
Policy on Antitrust
OMA Appendix G ‐ Use of AOAC Voluntary Consensus Standards to Evaluate Characteristics of a Method of Analysis
Expert Review Panel Policies and Procedures
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Road to First Action OMA Status
1. PTM – OMA Methods 2. Other Sole Source Methods 3. Response to Call for Methods
Method submitted
Expert Review Panels review all methods submitted methods
Notify Method author
Reject
ERP
Adopt
Published First Action OMA
Road to Final Action OMA Status
Method reproducibility must be demonstrated before Final Action consideration.
ERP determines if sufficient evidence merits a recommendation for Final Action status or repeal. • Only the OMB promotes a method to “Final Action” status or repeal the method. • Methods that did not meet the bar would be repealed. • Same for all method submissions
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PTM Overview for PTM‐OMA Harmonized Process • Administered by the Research Institute in 2003. • Well established and streamlined • Original approved by consensus with the OAs, OMB, RI Board of Directors and AOAC INTERNATIONAL Board of Directors. • ERP may be formed during Consulting Service. • Criterion for OMA: manufacturer’s method claims.
Recruiting Experts and Methods • AOAC issues – Call for Methods (Stakeholder affiliated methods) – Call for Experts • Sole Source/Individual Method Submissions – Individually completed Application not associated with an open Call for Methods
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Qualifications for ERP Membership Candidate must meet one of the following: • Demonstrated knowledge in the appropriate scientific disciplines. • Demonstrated knowledge regarding data relevant to adequate method performance.
• Demonstrated knowledge of practical application of analytical methods to bona fide diagnostic requirements.
Candidate application package includes: • Statement of Expertise • Current Abridged CV or Resume
ERP Member Vetting Process
Approved roster sent to AOAC President for volunteer appointment
Candidate submits application package
Reviewed by AOAC staff with recommendation to OMB
Reviewed by OMB and roster approved
• All members serve at the pleasure of the AOAC President • OMB assigns a representative to serve as a resource for every ERP
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Candidate Method Assignments A minimum of primary and secondary reviewers may be assigned to every method. In depth review via review form Prepare to attend and speak on the method and make a recommendation for ERP discussion and consideration. Review forms are completed and returned to AOAC staff in advance of the meeting. An email is sent with information on how to access the candidate methods and how to submit reviews
Members of both Committee on Safety and Committee on Statistics serve as advisory resources for all ERPs
Candidate Method Reviews
In your judgment, does the method sufficiently meet the Standard Method Performance Requirements (SMPR) or community‐based guidance?
In your judgment, is the method scientifically sound and can be followed? In your judgment, what are the strengths and weaknesses of the method? In your judgment, how do the weaknesses weigh in your recommendation for the method? In your judgment, will the method serve well the stakeholder community that will use the method? In your judgment, what additional information may be needed to further support the method meeting the SMPR or community‐based guidance? Members of both Committee on Safety and Committee on Statistics serve as advisory resources for all ERPs
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ERP Meetings ERPs can meet in person at a minimum of twice a year and up to four times per year*: AOAC Mid‐Year meeting (DC metro area) AOAC Annual Meeting. *2 additional designated times for proprietary method Organziational Affiliates At the ERP meeting: Reviews will be presented and the reviewers can make a motion to the ERP whether to adopt the method as First Action OMA. ERP discusses the method. ERP renders a decision on First Action status. ERP renders decisions on modifications to Official Methods . If the method is adopted ERP decides on what additional information is needed to recommend the method for Final Action status
ERP Teleconferences • Only after the initial in‐person ERP meeting for First Action consideration of methods • Possible for some method modifications • Possible for First Action to Final Action ERP recommendations
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ERP Meetings
Quorum
Presence of 7 vetted ERP members
Presence of 2/3 vetted ERP members
OR
WHICHEVER IS GREATER IF NO QUORUM, NO OFFICIAL MEETING
Method Review Overview
Method authors may be invited to make a presentation on their method REVIEWERS PRESENT THEIR REVIEWS AND MAY INITIATE A MOTION TO ADOPT THE METHOD IF THEY CHOOSE Chair recognizes each reviewer Reviews are presented.
If in favor, reviewers may make and second a motion to adopt adopt the method Chair can then entertain discussion on themethod Chair can call for a vote once deliberation is complete
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Consensus – First Action Adoption
First Action Official Methods status is granted:
Method must be adopted by unanimous decision of ERP on first ballot, if not unanimous, negative votes must be based on scientific reasons.
Negative voter(s) can be overridden by 2/3 of voting ERP members after due consideration.
Method becomes First Action on the date when ERP decision is made.
Consensus – First Action to Final Action
The ERP may then reach consensus on any additional information that it needs to review to be able to make a recommendation for Final Action Official Methods status.
This is a separate motion.
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ERP Meetings – Review for First Action METHOD AUTHOR: present any method and any resulting changes to the method since submission for review, summary of SLV and/or reproducibility evaluation, any recognitions (from AOAC or external) and, final draft of method proposed for decision
ERP CHAIR & MEMBERS: present reviews and discuss any resulting issues or questions on the method, review and agree upon final draft of method proposed for decision, and chair calls for ERP decision in accordance to procedures.
CONSENSUS: Method must be adopted by unanimous decision of ERP on first ballot. If not unanimous, negative votes must delineate scientific reasons. Negative voter(s) can be overridden by 2/3 of non‐ negative voting ERP members after due consideration. Abstentions do not count towards vote; in case of multiple abstentions the results will need to be evaluated. Staff will monitor and record consensus voting.
STAFF: Will organize and coordinate meeting, record ERP actions and decisions, draft ERP report and distribute after chair approval, work with chair and OMB liaison to complete checklist and assemble recommendation package for OMB.
ERP Methods Review & Approval
Methods should be scientifically sound with demonstrating that it will meet the needs of those using the method (evidenced by meeting the standard, or other acceptance criteria)
ERPs have approved methods with evidence of high potential to First Action and request additional work or support be submitted for review prior to ERP convening to recommend an action to OMB
OMB requires a justification or rationale for methods that are deemed acceptable and adopted but may not fully meet the standard set or acceptance criteria.
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OMB Expectations for First Action
Safety review needed prior to First Action status
SLV type of supporting information available per the SMPR
• Applicability, Method Performance Requirements Table, System Suitability, Reference Materials, and Validation Guidance
• Documented method performance versus a SMPR • Document reasons for acceptability if method does not meet the SMPR
Comparison to SMPR
Any approved method(s) along with supporting manuscript(s) and documentation sent to AOAC Publications after the meeting.
Method incorporating ERP revisions (preferably in AOAC Format) Method Manuscript incorporating specified ERP revisions (in AOAC Format) Signed AOAC Copyright Authorization form
NO OMA NUMBER ASSIGNED UNTIL ALL DOCUMENTATION SUBMITTED
Publication of First Action Methods
Method and method manuscript prepared for publication in the Official Methods of Analysis of AOAC INTERNATIONAL and in Journal of AOAC INTERNATIONAL Updates on methods approved or status changes are published in the Inside Laboratory Management magazine and on the AOAC website
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ERP Meetings – Method Tracking METHOD AUTHOR: present any method feedback obtained and any resulting changes to the method, any reproducibility information, any implemented ERP recommendations, final draft of method proposed for decision ERP MEMBERS: present any method feedback obtained and
discuss any resulting changes to the method, any reproducibility information, any implemented ERP recommendations, review and agree upon final draft of method proposed for decision, and make a recommendation to OMB. CONSENSUS: 2/3 vote in favor of a motion. Abstentions do not count towards vote; in case of multiple abstentions. Staff will monitor and record consensus voting.
STAFF: Will organize and coordinate meeting, record ERP actions and decisions, draft ERP report and distribute after chair approval, work with chair and OMB liaison to complete checklist and assemble recommendation package for OMB.
Documentation Needed
Method Safety Evaluation
Reference Materials
Evidence of Single Laboratory Validation or equivalent
Evidence of Reproducibility Assessment
Published First Action OMA
Method Performance versus SMPR or acceptance criteria
Final draft of First Action OMA to be considered for status update
Rationale or Justification for Repeal or Continuance of First Action OMA
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OMB Meeting for Review of ERP Recommendations
OMB Review (renders decision on recommendation)
ERP Chair/or designee (addresses questions/comment)
OMB Liaison (presents recommendation)
Modifications to Official Methods • Types of Modifications – Editorial
– Major – Minor
• Applicable to First Action and Final Action OMA
• Relevant to all ERPs
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Editorial Modifications • The applicant must submit a written explanation of the change(s) including a statement that the modification does not alter the validated performance of the method.
• Examples include: Typos or editorial corrections or clarifications that strengthen instruction.
• Methods that have undergone an editorial modification will retain the same number.
Editorial Changes
• Editorial changes to methods only require AOAC staff review and the change is made to the OMA with changes noted in next printed edition of OMA. • A list of the methods with editorial modifications will be published in Inside Laboratory Management and on the Website.
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Minor Modifications • Results in no changes to the current validated performance. There is no significant effect to the results. The method will retain the original number. • Supporting data to justify the proposed modification must be submitted. Equivalency data is required unless adequate Justification to exclude this data is provided. • Examples include: Reagent change, a change in a column or consumables that do not impact the validated method performance.
Major Modifications • Results in a change to the current validated performance of the method. • This level of modification will result in a new method as part of AOAC standards development and will receive a new method number. • Examples include: significant change to the technology, sample preparation, or chemistry.
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Minor & Major Modifications
Based on AOAC staff review, a public comment period for the proposed modification is required.
Applicant Options
• Following the comment period, any comments are reconciled and recommends a response to the applicant. • The applicant can decide to proceed based on the reconciled comments
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Pathways for Minor & Major Modification • If applicant decides to
proceed, an ERP is formed – Level of modification determined by ERP
– Applies to
modifications of First Action and Final Action methods
Documentation and Communication • AOAC carefully documents the actions of Stakeholder Panel, Working Groups, and Expert Review Panels • AOAC will prepare summaries of the meetings – Communicate summaries to the stakeholders – Publish summaries in the Referee section of AOAC’s Inside Laboratory Management • AOAC publishes its voluntary consensus standards and Official Methods – Official Methods of Analysis of AOAC INTERNATIONAL – Journal of AOAC INTERNATIONAL • AOAC publishes the status of standards and methods in the Referee section of AOAC’s Inside Laboratory Management
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Requirements for ERP Service
Must have demonstrated expertise in the method, technology, analyte/matrix, etc… Be a subject matter expert. Must be able to attend ERP meetings Must be able to complete assigned reviews on time Must be prepared to speak on the method and share reviews during the meeting Must be proactive in tracking assigned First Action Official Methods Must be able to assist in peer reviewing paper for publication Must sign and submit AOAC Volunteer Acceptance Form
General Expectations for ERPs • You can expect to have a minimum of three weeks to review methods prior to ERP meeting. – You are requested to submit written reviews by specified deadline. Please alert staff if you are not able to complete on time. – You may have individually assigned methods to review or all of the methods to review. Please be prepared to discuss these methods during meeting. – You may use the OMA appendices as guidance for types of validation work that can be expected. If additional information is needed, please ask staff. • ERP Meeting Quorum – If there is no quorum, there is no official meeting. Please alert staff as early as possible if you are not able to attend a meeting. • ERP Consensus – ERP consensus may not reflect your own personal view – There may be times when a method may not meet all of the criteria exactly; however, the ERP can adopt the method.
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Ethical Expectations of AOAC Expert Review Panel Members • Respect for your peer ERP members and chair – Each member has been vetted for expertise relevant to the review of the method(s) in the ERP • Be considerate of each others perspectives and points of view • Be considerate of the ERP’s consensus even if you disagree – Inform staff as early as possible if you cannot attend the scheduled ERP meeting • Be considerate in that your absence can impact the quorum of the ERP and its ability to have an official meeting to make decisions – Notify staff and/or disclose in the ERP meeting if you have a direct or perceived conflict of interest for a specific method • Please review AOAC’s policy on Volunteer Conflict of Interest Ethical Expectations of Expert Review Panel Members (con’t) • Respect for Method Authors and Intellectual Property – Each Method Author is encouraged to attend the ERP meeting – Each candidate methods (not yet adopted or published as Official Methods of Analysis of AOAC INTERNATIONAL ) are still the intellectual property of the method author. Therefore, the information is shared only with the vetted ERP members and is available during the meetings. Please do not distribute the information without expressed written permission from an appropriate AOAC staff liaison. – Be clear about and justify how additional recommended work is a requirement for First Action, a requirement for Final Action consideration, or something recommended, but not necessary. – Keep your focus on the science
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ERP Chair Responsibilities
Before Meeting
During Meeting
Moderate discussions based on agenda
Work with staff on meeting coordination
Engage staff to encourage members to reach decision points
Review submitted and/or assigned methods
Engage staff on procedural questions
Review method reviews if applicable
Engage discussion on feedback mechanism
Review SMPR(s) and/or relevant guidance and criteria
ERP Chair Responsibilities
Other Efforts and Recognitions Can nominate methods for OMB Award
After Meeting Review Meeting Report and Approve Final Version
Can nominate ERP members for OMB Award
Assist with any follow up on methods
Can assist in identifying methods for review
Assist in Publication Reviews
Can serve as a guest editor for the Journal
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Roles and Responsibilities
AOAC Official Methods Board Vet and approve stakeholder panel chair & voting members Vet and approve ERP membership and AOAC Experts Render decisions on status of First Action methods (Final Action, repeal, etc…) Assign a liaison to each stakeholder panel and ERP Coordinate OMB Awards AOAC Expert Review Panels Review methods and meet in person to render decisions on methods for First Action Official Methods SM status. Track First Action Official Methods SM and modify, if necessary Recommend First Action methods after 2 years or less to OMB for Final Action, continuance, or Repeal Participate in Consulting Service and PTM reviews for OMA and harmonized PTM and harmonized OMA method studies AOAC Experts Review and approve PTM validationtesting protocol documentation Peer review of PTM validation manuscript and supporting documentation AOAC Research Institute ‐ PTM Expert Reviewers Peer Review of PTM validationmanuscripts and supporting documentation
AOAC Research Institute Independent Laboratories Conduct independent evaluation of candidate method using AOAC approved testing protocols AOAC Stakeholder Panels Develop voluntary consensus standards Assign working groups to draft standards method performance requirements Voting members demonstrate consensus on behalf of stakeholders AOAC Staff Coordinate method reviews and method approval activities Coordinate OMB meetings Provide trainings and orientations Maintain website and communication Document and publish actions and decisions Coordinate standards development activities Publish standards and methods AOAC Research Institute Technical Consultants Draft validation protocols in Consulting Service for assigned methods
Facilitate PTM evaluation of assigned candidate methods Facilitate comments/responses for assigned OMA reviews
Questions?
Thank you
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AOAC Official Methods of Analysis SM (OMA) EXPERT REVIEW PANEL FOR GLUTEN ASSAYS
TABLE OF CONTENTS A. AOAC FINAL ACTION REQUIREMENTS FOR FIRST ACTION OFFICIAL METHODS – COVER SHEET I. AOAC OFFICIAL METHOD 2012.01: GLIADIN AS A MEASURE OF GLUTEN IN FOODS CONTAINING WHEAT, RYE, AND BARLEY, ENZYME IMMUNOASSAY METHOD BASED ON A SPECIFIC MONOCLONAL ANTIBODY TO THE POTENTIALLY CELIAC TOXIC AMINO ACID PROLAMINE SEQUENCES, FIRST ACTION 2012 .......................................................... 3 A. METHOD FEEDBACK.......................................................................................... 6 B. ARTICLE: GLIADIN AS A MEASURE OF GLUTEN IN FOODS CONTAINING WHEAT, RYE, AND BARLEY—ENZYME IMMUNOASSAY METHOD BASED ON A SPECIFIC MONOCLONAL ANTIBODY TO THE POTENTIALLY CELIAC TOXIC AMINO ACID PROLAMIN SEQUENCES: COLLABORATIVE STUDY........................ 16 C. EXPERT REVIEW PANEL REPORT (SEPTEMBER, 2016)........................................ 24 D. AOAC PERFORMANCE TESTED CERTIFICATE #120601 .......................................... I. REFERENCE AND GUIDANCE DOCUMENTATION ( Access via the link below ) In addition to all AOAC Official Methods Board Guidance regarding AOAC First Action and AOAC Final Action Official Methods status. 1. APPENDIX D : GUIDELINES FOR COLLABORATIVE STUDY PROCEDURES TO VALIDATE CHARACTERISTICS OF A METHOD OF ANALYSIS 2. APPENDIX M: VALIDATION PROCEDURES FOR QUANTITATIVE FOOD ALLERGEN ELISA METHODS: COMMUNITY GUIDANCE AND BEST PRACTICES 3. APPENDIX N: ISPAM GUIDELINES FOR VALIDATION OF QUALITATIVE BINARY CHEMISTRY METHODS
CHECKLIST FOR FINAL ACTION METHOD RECOMMENDATION 2012.01: GLIADIN AS A MEASURE OF GLUTEN IN FOODS CONTAINING WHEAT, RYE, AND BARLEY, ENZYME IMMUNOASSAY METHOD BASED ON A SPECIFIC MONOCLONAL ANTIBODY TO THE POTENTIALLY CELIAC TOXIC AMINO ACID PROLAMINE SEQUENCES, FIRST ACTION 2012 OMB GUIDANCE FOR AOAC ERPS Considered? Comments Method Applicability Yes The determination of gluten in rice flour and rice
based unprocessed and processed foods as specified in the applicability statement. All addressed prior to First Action. Reference materials are addressed in the protocols for SLV. AOAC Performance Tested SM cert.: 120601; Harmonized PTM‐OMA
Safety Concerns Reference Materials
Yes No
Single Laboratory Validation
Yes
Immer et. al., J. AOAC Int . 95 , 1118 (2012)
Reproducibility/Uncertainty and Probability of Detection
Yes
Comparison to SMPR
No
SMPR, not applicable. This method was validated by previous AOAC Committee D/E prior to the establishment of the Expert Review Panels.
Feedback from Users of Method
Yes
Discussed in ERP meeting
ERP Recommendation to Repeal First Action Method
Not Applicable
DOCUMENTATION Safety Evaluation Reference Materials
Available?
Comments
Yes No Yes Yes Yes Yes No Yes Yes Yes Yes
Completed prior to AOAC First Action
In manuscript
SLV or PTMs
AOAC PTM cert.: 121301; Harmonized PTM‐OMA Precollaborative, Independent, Collaborative
Approved Validation Protocols
Statistics Review
Completed prior to AOAC First Action
Method Published in OMA
2014.03
Method Performance vs SMPR criteria
SMPR, not applicable. This method was validated using the guidance of OMA Appendices D, M, and N. Discussed in ERP meeting
Feedback Information Additional Recognition(s)
AOAC PTM cert.: 120601
ERP Reports
September, 2016
Immer et. al., J. AOAC Int . 95 , 1118 (2012)
Manuscript(s) Published in JAOAC
Method Recommended for Final Action
Yes
The OMA title was changed to “Gliadin as a Measure of Gluten in Rice and Corn Based Foods” in lieu of “Gliadin as a Measure of Gluten in Foods Containing Wheat, Rye, and Barley”
of sample extracts is compared with response observed with calibrators. B. Apparatus Apparatus specified has been tested. Equivalent apparatus may be used. ( a ) Grindomix GM 200 .—For sample homogenization (Retsch GmbH, Haar, Germany). ( b ) Water bath .—Gesellschaft für Labortechnik mbH (Burgwedel, Germany). ( c ) Bench-top centrifuge .—Multifuge 3L-R, operating at 2500 rpm (Thermo Electron GmbH, Dreieich, Germany). ( d ) Glass tubes .—10 mL; for extraction (Brand GmbH, Wertheim, Germany). ( e ) Polystyrol tubes .—5 mL; for sample dilution (Sarstedt, Nümbrecht, Germany). ( f ) Microtiter plate reader .—With 450 nm filter (Tecan Deutschland GmbH, Crailsheim, Germany). ( g ) Micropipet .—Accurately delivering 100 µL ± 1%. ( h ) Glassware .—Wash bottle (1000 mL) and graduated cylinders. ( i ) Rotator 3100 CMV or equivalent.— Fröbel Labortechnik (Lindau, Germany). C. Antibody Characteristics Antibodies must satisfy the following criteria: ( 1 ) Bind to gliadin derived from wheat and to related prolamins derived from rye and barley ( 2 ) Recognize the potential celiac toxic structure QQPFP and related sequences ( 3 ) Bind to the a -, b -, γ-, and Ω-gliadin motifs in nonheated and heated food, extracted by cocktail solution ( 4 ) No binding to- oats, maize, rice, teff, buckwheat, quinoa, and amaranth ( 5 ) Bind with high affinity to allow an LOD of 1.5 mg/kg gliadin or related prolamins ( 6 ) Able to build a stable POD labeled conjugate, stable for more than 1 year
AOAC Official Method 2012.01 Gliadin as a Measure of Gluten in Rice- and Corn-Based Foods Enzyme Immunoassay Method Based on a Specific Monoclonal Antibody to the Potentially Celiac Toxic Amino Acid Prolamine Sequences First Action 2012 Final Action 2016 Caution : Cocktail solution necessary for sample preparation contains β -mercaptoethanol. Use a chemical hood for sample preparation. Stop solution contains 1 M sulfuric acid. Avoid skin and eye contact ( see Material Safety Data Sheet). See Table 2012.01 for the results of the interlaboratory study supporting acceptance of the method. A. Principle The method is based on an enzyme immunoassay format using a monoclonal antibody that can determine gliadin derived from wheat and related prolamins derived from rye and barley. The antibody binds to the potentially celiac toxic amino acid sequence QQPFP (1) and to related sequences, which exist as motifs on all the gliadin subunits. The antibody detects prolamins in nonheated and heated food by using an additional specific extraction method (cocktail solution). No cross-reactivity exists to oats, maize, rice, millet, teff, buckwheat, quinoa, and amaranth. Prolamins from food are extracted by using a cocktail solution containing β -mercaptoethanol and guanidine hydrochloride described by García et al. (2), following an extraction with 80% ethanol. After centrifugation, the supernatant is used in a second- step sandwich method. The analyte is incubated in monoclonal antibody-coated wells forming an antibody–antigen complex. In a second step, an antibody peroxidase (POD) conjugate reacts with the complex to form an antibody–analyte–antibody complex. A chromogen/substrate reaction with the immobilized POD labeled conjugate determines the bound analyte. Nonimmobilized components are removed by washing between steps. The response
Table 2012.01. Interlaboratory study results for gliadin by RIDASCREEN ® Gliadin Material No. of labs (outliers) Mean, mg/kg Recovery, % Matrix Level, mg/kg
RSD r
, %
RSD
, %
R
Maize Maize Maize Maize Rice Rice a Rice
168
19 (1) 20 (0) 18 (2) 20 (0) 18 (2) 17 (1) 20 (0) 20 (0) 17 (0)
141.8
84.4
20.8 37.7 14.2 32.0 18.3 26.8 26.8 37.4 29.7
28.6 40.3 32.4 41.5 25.6 35.4 40.7 38.1 52.1
35 79
36.8 74.1
105.0
93.8
0
8.3
41
34.7 <1.5
84.6
0
147
126.6
86.1 89.3
Wheat starch
14 13
12.5 14.1 13.2 <1.5 <1.5
Rice flour
108.5
Wheat starch Maize flour a Maize flour a
13.5 <1.5 <1.5
97.8
a Negative samples were not included in the statistical evaluation.
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and measure in a microtiter plate reader at 450 nm vs air within 30 min after stopping the reaction. Do not reuse wells of the plate. Use separate pipet tips for each standard and each sample extract to avoid cross-contamination. Use a multistepper pipet for adding the conjugate, substrate/ chromogen, and stop solution. Use a single tip for each of these components. Components and procedures of test kit have been standardized for use in this procedure. Do not interchange individual components between kits of different batches (lot numbers). Do not freeze any of the kit components. Carefully dilute the components that are included in the kit as concentrates; avoid contaminations by airborne cereal dust or dirty laboratory equipment. Wear gloves during preparation and performance of the assay. Clean surfaces, glass vials, mincers, and other equipment with 60% ethanol. Carry out sample preparation in a room isolated from ELISA procedure. Check for prolamin contaminations of reagents and equipment. F. Preparation of Test Samples Weigh 5 g sample and grind to a powder as fine as possible to obtain maximal surface. Weigh 0.25 g of the solid ground sample or use 0.25 mL of a liquid sample in a 10 mL glass vial and add 2.5 mL cocktail. Close vial and mix well (avoid cross-contamination). If tannin- and polyphenol-containing samples (e.g., chocolate, chestnut, or buckwheat) are prepared, add an additional 0.25 g skim milk powder (food quality) to the sample–cocktail solution. Incubate for 40 min at 50°C (122°F) in a water bath. Let sample cool down; then mix with 7.5 mL 80% ethanol. Close vial and shake for 1 h upside down or by a rotator at room temperature (20–25°C/68–77°F). Centrifuge 10 min at 2500 g at room temperature (20–25°C/68–77°F). Remove the supernatant (extract) in a screw-top vial and keep for testing. Dilute the sample at least 1:12.5 (1 + 11.5, 0.1 + 1.15 mL) with the prepared sample dilution buffer (depending on the expected prolamin content of the sample). Dilute serially from the first dilution, if necessary mixing thoroughly each time before diluting further. Use 100 µL per well in the assay. G. Preparation of Components Delivered with Kit ( a ) Sample diluent . — Provided as a concentrate (5-fold). Only the amount which is actually needed should be diluted 1:5 (1 + 4) with distilled water (e.g., 3 mL concentrate + 12 mL distilled water, sufficient for the dilution of 10 samples). Make sure that the buffer is not contaminated with gliadin. ( b ) Antibody enzyme conjugate.— (Bottle with red cap.) Provided as a concentrate (11-fold). Since the diluted enzyme conjugate solution has a limited stability, only the amount that is actually needed should be diluted. Before pipetting, the conjugate concentrate should be shaken carefully. For reconstitution, the conjugate concentrate is diluted 1:11 (1 + 10) with distilled water (e.g., 200 μL concentrate + 2.0 mL distilled water, sufficient for two microtiter strips). Take care that the water is not contaminated with gliadin. ( c ) Washing buffer .—Provided as a 10-fold concentrate. Before use, the buffer must be diluted 1:10 (1 + 9) with distilled water (i.e., 100 mL buffer concentrate + 900 mL distilled water). Prior to dilution, dissolve any crystals formed by incubating the buffer in a water bath at 37°C (99°F). The diluted buffer is stable at 2–8°C (35–46°F) for 4 weeks.
( 7 ) Show reproducible affinity, sensitivity, specificity, and stability from batch to batch for more than 1 year ( 8 ) Monoclonal antibodies are preferred; polyclonal antibodies can be used if they fulfill the same specificity criteria to react with wheat, rye, and barley to 100% and have no cross-reactivity to oat, maize, teff, and others D. Reagents Items ( a )–( i ) are available as a test kit (RIDASCREEN® Gliadin; R-Biopharm AG, Darmstadt, Germany). All reagents are stable for 18 months from date of manufacture at 2–8°C (36–46°F). Refer to kit label for current expiration. Equivalent antibodies may be used for ( a ) and ( c ) provided they satisfy characteristic criteria in C . ( a ) Antibody-coated microwell strips .—Monoclonal antibodies are coated in 20 mM phosphate buffered saline (PBS), pH 6.0, onto a set of twelve 8-microwell strips (NUNC, Roskilde, Denmark), containing 0.01% sodium azide as preservative. ( b ) Wash buffer concentrate .—100 mL/bottle, 10x concentrate. Contains a final concentration of 20 mM PBS (0.9% sodium chloride) with 0.1% Synperonic and 0.01% bronidox L as preservative. ( c ) Peroxidase-labeled antibody. —One vial (1.2 mL, 11x concentrated). ( d ) Gliadin ready-to-use standards (antigen). —Six vials (1.3mL each, ready to use). Prepared by Sigma gliadin or own preparation, dissolved in 60% ethanol at a concentration of 1 mg/mL. Solution is further diluted in 20 mM PBS–Tween (0.9% sodium chloride, 0.05% Tween 20) containing 0.22% fish gelatin (Sigma) to 0, 5, 10, 20, 40, and 80 ng/mL gliadin, calibrated to the Working Group on Prolamin Analysis and Toxicity (WGPAT) gliadin (86% highly purified gliadin from 40 different European wheat varieties). ( e ) Substrate .—One vial, 7 mL (urea peroxide). ( f ) Chromogen .—One vial, 7 mL (tetramethylbenzidine in methanol). Can be added either separately or mixed 1 + 1 with ( e ) before pipetting. ( g ) Stop solution .—One vial, 14 mL (1 N H 2 SO 4 ). ( h ) Sample dilution buffer .—60 mL, 5x concentrate. Contains a final concentration of 20 mM PBS–Tween (0.9% sodium chloride, 0.05% Tween 20) with 0.22% fish gelatin (Sigma) and 0.01% Kathon as preservative. ( i ) Cocktail solution .—One vial, 105 mL. Recommended but not provided with the test kit: ( a ) Skim milk powder.— Food quality. ( b ) Samples .—Three control samples (powder), one nongliadin- containing sample (rice flour) and two prolamine-contaminated maize samples (A and B, concentration given by a certificate), which can be extracted with 60% ethanol and diluted further with the sample dilution buffer to control the test from run to run. E. General Instructions Store kit at 2–8°C (35–46°F). Let all kit components come to 20–25°C (68–77°F) before use. Return any unused microwells to their original foil bag, reseal them together with the desiccant provided, and store at 2–8°C (35– 46°F). The colorless chromogen is light-sensitive; therefore avoid exposure to direct light. Include ready-to-use standards in duplicates to each run of diluted sample extracts in duplicates. Add the diluted antibody– POD conjugate (diluted by water) to all wells. Add substrate and chromogen simultaneously. Stop the reaction with stop solution,
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Gluten content of a sample can be calculated from the gliadin value, as gliadin generally represents 50% of the proteins present in gluten. Gluten values can be expressed in mg/kg by multiplying the gliadin value by 2. Example calculation : A sample was extracted with the recommended dilution factor of 500. The absorbance value of the sample corresponds to 10 ng/mL gliadin in the calibration curve. By multiplying the obtained value by the factor 500 leads to 5000 ng/mL, corresponding to 5 mg/kg gliadin, respectively, 0.0005% gliadin. To calculate the gluten content, multiply by factor 2, which results in 10 mg/kg gluten, respectively, 0.001% gluten. This sample is considered to be gluten-free because the gluten concentration is below 20 mg/kg gluten. LOD was calculated by testing 10 blank samples/matrix; mean values and standard deviation (SD) were calculated. LOD was defined as mean + 3x SD. LOQ was verified by analyzing 10 replicates of a food sample, which contains a gliadin content close to standard 2 [5 ng/mL × 500 (dilution factor) = 2.5 ppm gliadin]. In parallel standard 1 (= 0 ng/mL gliadin) was measured 10 times. The variation of standard 1 (absorbance value + 3x SD) was confirmed. The mean value – 3x SD was found significantly different from zero in consideration of the CV. K. Criteria for Acceptance of Standard Curve The course of the calibration curve is shown in the Quality Assurance Certificate, enclosed in the test kit. In comparison with the certificate, higher values of the absorbance at 450 nm, especially for the zero calibrator, may be a result of insufficient washing or gliadin contamination. A further dilution and repeated measurement of the samples is recommended for absorbance values (450 nm) higher than standard 6. This additional dilution factor must be taken into consideration during calculation. Indication of instability or deterioration of reagents is shown by any coloration of the chromogen solution prior to test implementation or if values of less than 0.6 absorbance units for standard 6 occur. SD of replicates should be less than 10%. Test controls offered by R-Biopharm should be measured in the reported ranges from run to run. References: (1) Osman, A.A., Uhlig, H.H., Valdes, I., Amin, M., Mendez, E., & Mothes, T. (2001) Eur. J. Gastroenterol. Hepatol . 13 , 1189–1193 (2) García, E., Llorente, M., Hernando, A., Kieffer, R., Wieser, H., & Méndez, E. (2005) Eur. J. Gastroenterol. Hepatol . 17 , 529–539 J. AOAC Int . 95 , 1118(2012) Revised 2016 to update title as part of Final Action vote (change “foods containing wheat, rye, and barley” to “rice- and corn- based foods”); March 2017 to include modification of the wash solution to substitute thimerosal in the washing buffer by the mercury-free preserving agent bronidox L, D ( b ). Posted: July 6, 2017
H. Determination Bring all reagents to room temperature (20–25°C/68–77°F) before use. Do not allow microwells to dry between working steps. Insert a sufficient number of wells into the microwell holder for all standards and samples to be run. Record standard and sample positions. Add 100 µL of each standard solution or prepared sample to separate wells, mix 10 s manually, and incubate for 30 min at room temperature (20–25°C/68–77°F). Dump the liquid out of the wells, and then tap the microwell holder upside down vigorously (three times in a row) against absorbent paper to ensure complete removal of liquid from the wells. Fill all the wells with 250 µL diluted washing buffer and dump out the liquid again. Repeat two more times. Add 100 µL of the finally diluted enzyme-labeled conjugate to each well, mix 10 s manually, and incubate for 30 min at room temperature (20–25°C/68–77°F). Dump the liquid out of the wells, and then tap the microwell holder upside down vigorously (three times in a row) against absorbent paper to ensure complete removal of liquid from the wells. Fill all the wells with 250 µL diluted washing buffer and dump out the liquid again. Repeat two more times. Add 50 µL substrate and 50 µL chromogen to each well. Mix gently by shaking the plate 10 s manually and incubate for 30 min at room temperature (20–25°C/68–77°F) in the dark. Positive wells should develop a blue color, indicating the presence of prolamins. Add 100 µL stop reagent to each well. Mix gently by shaking the plate manually. The color of positive prolamin-containing wells changes from blue to yellow. I. Reading Read the results with a microtiter plate reader. Measure the absorbance at 450 nm. Read within 30 min against air after addition Determine the gliadin content of each set of duplicate sample wells by reference to a calibration curve measured by the actual test run utilizing special computer software or semilogarithmic paper; plot absorbance of standards (linear scale) vs gliadin content of standards (logarithmic scale). The standard calibration curve of the ELISA covers a range from 2.5 to 40 mg gliadin/kg sample, which corresponds to a range of 5–80 ng/mL gliadin in the calibrators. Convert the units ng gliadin/mL diluted sample to mg gliadin/kg sample as follows: Multiply the amount in ng/mL by the dilution factor. Divide the product by 1000 to achieve units of mg/kg. The dilution factor corresponds to the sample preparation and is usually 500; however, 1000 was used in this study. Absorbance below standard 2 (5 ng/mL gliadin) implies that the sample assayed is diluted too much or that no gliadin or gliadin below the LOQ is present in the sample. of stop solution. J. Calculations
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