AOAC CASP Meeting

AOACTHANKS OUR CASP MEMBERS CASP Pioneers :

• ABCTesting, Inc. • Association of Food and Drug Officials • Bia Diagnostics • BioRad

• PathogenDx • PerkinElmer • R‐BiopharmAG • SCIEX • Supra Research & Development • TEQAnalytical

• BIOTECON Diagnostics • GW Pharmaceuticals  • Industrial Laboratories  • Materia Medica Labs • MilliporeSigma • SōRSETechnology

• TITANAnalytical • Trilogy Analytical

CASP Affiliates: • Charm Sciences • Crystal Diagnostics • Hygiena • Institute of FoodTechnologists • BIOTECON • Lazarus Naturals • Medicinal Genomics • SC Labs • Emerald Scientific

CASP Partners • CEMCorporation • CV Sciences • Eurofins Scientific • Trace Analytics 

AOAC INTERNATIONAL Cannabis Analytical Science Program (CASP) Saturday, September 7, 2019 | 9:00AM – 5:00PM EDT Sheraton Denver Downtown, Grand Ballroom 1

MEETING AGENDA

I. WELCOME, INTRODUCTIONS AND ANNOUNCEMENTS (9:00AM – 9:15AM) David Schmidt, AOAC INTERNATIONAL II. PROGRAM OVERVIEW (9:15AM – 9:45AM) Scott Coates, AOAC INTERNATIONAL III. CONSENSUS BUILDING AT AOAC INTERNATIONAL (9:45AM – 10:15AM) Deborah McKenzie, AOAC INTERNATIONAL

IV. REPORT FROM THE MICROBIAL CONTAMINANTS WORKING GROUP (10:30AM – 11:30AM) Pat Bird, PMB Biotek and Julia Bramante, Colorado Department of Public Health & Environment V. REPORT FROM THE CHEMICAL CONTAMINANTS WORKING GROUP (11:30PM – 12:30PM) Susan Audino, Audino and Associates VI. REPORT FROM THE CANNABOIDS IN CONSUMABLES WORKING GROUP (1:30PM – 2:30PM) Holly Johnson, American Herbal Products Association VII. CHALLENGES IN CHEMICAL CONTAMINANT TESTING AND APHL’S CANNABIS COMMUNITY OF PRACTICE (2:30PM – 2:50PM) Heather Krug, Colorado Department of Public Health & Environment VIII. TESTING CBD PRODUCTS FOR SAFETY AND QUALITY (3:05PM – 3:25PM) Toby Astill, PerkinElmer IX. RESPONSIBLE MICROBIAL TESTING POLICY IN CANNABIS: CHALLENGES AND FUTURE OUTLOOK (3:25PM – 3:45PM) Ben Katchman, PathogenDx X. CBD/THC IN HEMP PERSPECTIVE (3:45PM – 4:05PM) Roy Bingham, BDS Analytics XI. UPDATE FROM INSTITUTE OF FOOD TECHNOLOGIES LEGALIZED HEMP AND EDIBLES (4:05PM – 4:25PM) Rosetta Newsome, Institute of Food Technologists XII. 2020 WORK PLAN PROPOSAL (4:25PM – 4:45PM) Scott Coates, AOAC INTERNATIONAL XIII. SUMMARY/WRAP UP (4:45PM – 5:00PM) Scott Coates & Palmer Orlandi, AOAC INTERNATIONAL

Breaks at 10:15am and 2:50pm, Lunch at 12:30pm 08-26-2019 Version 7 – Subject to Change Without Notice

September 7, 2019 AOAC CASP Meeting – Presenter Bios

AOAC INTERNATIONAL CASP MEETING – SPEAKER BIOS

Toby Astill, Ph.D.

PerkinElmer

Global Marketing Manager for Cannabis & Hemp

Dr. Toby Astill is PerkinElmer’s Global Market Manager for Cannabis & Hemp. Dr. Astill has a Ph.D. in Chemistry and has spent the last 15 years working in Science, Technology and Business Roles for leading technology companies. For the last 8 years he has worked for PerkinElmer, based in locations across the West Coast of Canada & the US. Dr. Astill is now focused on driving PerkinElmer's global cannabis and hemp business; including but not limited to new product development, market & industry research, collaborations with customers, application development and marketing initiatives.

September 7, 2019 AOAC CASP Meeting – Presenter Bios

Susan Audino, Ph.D

Audino & Associates

Owner

Dr. Susan Audino is a chemist/chemometrician and independent consultant to chemical and biological laboratories. On behalf of Accreditation Bodies, she assesses laboratories to and is an instructor for multiple ISO/IEC standards such as ISO 17025. She is recognized as a leader in the cannabis industry, focusing on consumer safety and sound science in the development of official and consensus analytical test methods. Susan serves as a scientific advisor to several scientific organizations, regulatory bodies, and sits on expert review panels. Dr. Audino has chaired the AOAC cannabis advisory panel and currently chairs the chemical contaminants working group, and is a board member for the Center for Research on Environmental Medicine. She has provided seminars, workshops, webinars, and facilitated symposia domestically and internationally.

September 7, 2019 AOAC CASP Meeting – Presenter Bios

Patrick M. Bird, M.S. PMB BioTek Consulting LLC Owner

Pat has been an active member of the food safety community for the last 15 years serving in numerous roles including manufacturing and laboratory management before starting his own consulting business in 2018. Pat spent extensive time as the supervisor of a R&D laboratory where he oversaw the development and validation of rapid diagnostic methods for microbiology, allergens and mycotoxins. As a consultant, Pat continues to work with method developers, laboratories and industry to optimize method performance and testing workflow. In December 2018, Pat joined AOAC INTERNATIONAL as a technical consultant, where he works on the development of validation outlines for the Performance Tested Methods SM (PTM) and Official Method of Analysis SM (OMA) processes. Pat has been an active member within AOAC INTERNATIONAL for over 12 years, serving on the AOAC Research Institute Board of Directors and as the microbiology community lead for AOAC’s Technical Programing Council. Pat has authored and co-authored over 40 peer reviewed publications and has presented over 15 OMA validations to the Microbiology Expert Review Panel as part of the OMA First Action submission process. Pat is a member of A2LA’s Laboratory Accreditation Council, and actively participates in many working groups including ASTM D37 on Cannabis , ASTM E35 on Pesticides, Antimicrobials and Alternative Control Agents , IAFP’s Applied Laboratory Methods working group on method validation and verification and two working groups in US ISO/TC34/SC9 Food Microbiology: WG3 for Method Validation and WG19 Guidelines for conducting challenge studies. Pat holds a BS in Microbiology from the Ohio State University in Columbus, Ohio and a MS in Food Safety from Michigan State University in East Lansing, MI.

September 7, 2019 AOAC CASP Meeting – Presenter Bios

Julia Bramante, Ph.D.

Colorado Department of Public Health and Environment Lead Scientist

Julia began her career in the cannabis industry in 2014 at Gobi Labs, one of the first cannabis testing facilities to open in Colorado. She then transitioned to the Colorado Department of Public Health and Environment’s Marijuana Reference Laboratory where she currently serves as Lead Scientist. Julia is also the Chair of the Cannabis Chemistry Subdivision of the American Chemical Society and the Co-Chair of the AOAC CASP Microbial Contaminants Working Group.

September 7, 2019 AOAC CASP Meeting – Presenter Bios

Scott Coates, M.S.

AOAC INTERNATIONAL Senior Director of the AOAC Research Institute

Scott was appointed as the Senior Director of the AOAC Research Institute on July 1, 2018. He is responsible for daily management of and business development for the AOAC Research Institute. Scott also serves as the Program Lead for the Cannabis Analytical Science Program. Scott served as the Chief Science Office from 2009 until June 2018. In this capacity, he served as the technical lead for many AOAC projects. Scott led the writing and development of Appendix F in the Official Methods of Analysis of AOAC INTERNATIONAL that describes validation requirements and the development of Standard Method Performance Requirements . Before joining AOAC, he worked for 10 years as the Operations Manager for an in-vitro diagnostic manufacturer making medical test kits such as Strep tests and specialized bacterial culture media. Scott holds a B.S. in Microbiology (1978) and a M.S. in Biotechnology Management (1994) from the University of Maryland. Contact Scott at: scoates@aoac.org , (301) 924-7077 x137

September 7, 2019 AOAC CASP Meeting – Presenter Bios

Benjamin Katchman, Ph.D.

PathogenDx Principal Scientist

Dr. Katchman leads the research and development of sample preparation and assay development at PathogenDx, a Scottsdale, AZ based company which provides disruptive DNA-based pathogen testing technology and solutions for the cannabis, botanical, food and agricultural industries. Previously, Dr. Katchman spent worked with various universities and small companies developing novel nucleic acid and proteomic approaches for biomarker discovery and molecular diagnostics. This includes the development of plate-based and microarray-based diagnostic systems for use in clinical laboratories and point-of-care diagnostics for Arizona State University, FlexBioTech, Inc., and Eccrine Systems, Inc. Prior to joining PathogenDx, Dr. Katchman was a Principle Investigator at Eccrine Systems, Inc. where he led all clinical research, regulatory pathway implementation, assay development, and biomarker studies. He was the co-founder of FlexBioTech, Inc., where he led the development of novel OLED-based molecular diagnostic assay platforms. Dr. Katchman received a B.S. in Microbiology, from Indiana University in 2005, then carried out graduate research with Prof. Douglas F Lake at Arizona State University where he was the first to identify QSOX1 as a proto-oncogene and novel tumor biomarker resulting in over $2M in funding and multiple patents. He performed his postdoctoral research with Prof. Karen S Anderson and Prof. Joshua LaBaer where he designed novel high-density protein microarrays and novel clinical and point of care diagnostics utilizing flexible OLED based electronics resulting in over $15M in funding, multiple patents and high-impact publications.

September 7, 2019 AOAC CASP Meeting – Presenter Bios

Holly Johnson, Ph.D.

American Herbal Products Association

Chief Science Officer

Holly E. Johnson Ph.D., is the Chief Science Officer for the American Herbal Products Association (AHPA). She previously served for three years as Laboratory Director for Alkemist Labs, an ISO 17025 accredited natural products testing lab specializing in botanical dietary supplements. Dr. Johnson took her Ph.D. in Pharmacognosy at the College of Pharmacy, University of Illinois – Chicago (UIC), under renowned Pharmacognosist and researcher Dr. Norman Farnsworth. Holly was awarded a National Institutes for Health (NIH) Fellowship and trained at the UIC/NIH Center for Botanical Dietary Supplements. She was a Postdoctoral Research Fellow at the Institute for EthnoMedicine studying the etiology of neurodegenerative disease, and also worked for Waters Corporation conducting technical training and regulatory consulting for pharmaceutical and supplements companies. She is currently a Research Associate with the National Tropical Botanical Garden and serves on AOAC working groups, stakeholders’ panels, and expert review panels for Foods and Dietary Supplements. She is a member of the United States Pharmacopeia’s (USP) Medical Cannabis Expert Panel, the Editorial Board of the Journal of AOAC International, and she serves on the Advisory Boards of the American Botanical Council and the American Herbal Pharmacopeia. Holly has over 20 years’ experience working with natural products & botanicals and spent many happy years conducting research on medicinal plants and giving courses at the University of Hawaii.

September 7, 2019 AOAC CASP Meeting – Presenter Bios

Heather Krug, M.S.

Colorado Department of Public Health and Environment State Marijuana Laboratory Sciences Program Manager

Heather Krug is the State Marijuana Laboratory Sciences Program Manager for the Colorado Department of Public Health and Environment. She oversees inspections of Colorado retail/medical marijuana laboratories for compliance and suitability for certification, and is responsible for the state Marijuana Reference Laboratory. Heather holds a Bachelor of Science degree in Biomedical Sciences and a Master of Science degree in Forensic Toxicology.

September 7, 2019 AOAC CASP Meeting – Presenter Bios  

Rosetta Newsome, Ph.D. 

Institute of Food Technologists  Director of Science, Policy & Scientific and  Regulatory Affairs

Rosetta Newsome, Ph.D., is Director of Science, Policy & Scientific and Regulatory Affairs at the Institute of Food Technologists (IFT). Dr. Newsome is involved in developing content generated by the Science & Policy Initiatives team and guiding and participating in IFT’s scientific and regulatory affairs. She also leads IFT’s engagement in Codex, serves as a scientific and technical resource, and provides guidance in government relations and public policy activities. She has substantial experience in grant and contract activities, and extensive experience working with a variety of entities. She is a member of the American Association for the Advancement of Science, American Chemical Society, International Association for Food Protection, and Sigma Xi, as well as IFT. Rosie received her bachelor’s degree from Kentucky State University and master’s and doctoral degrees, focused on food science, from the University of Kentucky. The Institute of Food Technologists (IFT) is a global organization of nearly 17,000 individual members from more than 100 countries committed to advancing the science of food. Since 1939, IFT has brought together the brightest minds in food science, technology and related professions from academia, government, and industry to solve the world’s greatest food challenges. IFT works to ensure that members have the resources they need to learn, grow, and advance the science of food as the population and the world evolve. IFT believes that science is essential to ensuring a global food supply that is sustainable, safe, nutritious, and accessible to all.

Scott Coates Senior Director

AOAC Research Institute AOAC INTERNATIONAL

September 7, 2019

Cannabis-Related AOAC Standard Method Performance Requirements (SMPR) • SMPR 2017.001 Cannabinoids in Cannabis Concentrates • SMPR 2017.002 Cannabinoids in Dried Plant Materials • SMPR 2017.019

Cannabinoids in Chocolate • SMPR 2018.011 Pesticides in Cannabis

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Cannabis-Related AOAC Official Methods of Analysis

AOAC Official Method 2018.10 Cannabinoid in Dried Flowers and Oil Liquid Chromatographic Method AOAC Official Method 2018.11 Quantitation of Cannabinoids in Cannabis Dried Plant Materials, Concentrates, and Oils using Liquid Chromatography–Diode Array Detection Technique with Optional Mass Spectrometric Detection

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Cannabis Analytical Science Program (CASP) • AOAC decided to create a full-time, dedicated program to cannabis analytical science in 2018. • CASP to include all of AOAC’s programs:

• SMPR • OMA • Performance Tested Methods

• Laboratory Proficiency Testing • Publications  (Journal) • Meetings

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CASP Objectives • Facilitate a forum where the science of cannabis analysis is discussed with international experts. • Facilitate the development and publication of cannabis and hemp- specific methods and standards. • Develop cannabis and hemp reference materials.

• Establish a cannabis and hemp proficiency testing program in accord with International Standards. • Provide analytical and laboratory management training, in particular ISO accreditation training. • Provide resources and education to regulators responsible for establishing rules and laws around cannabis and hemp.

Current CASP Projects Microbiology in Cannabis • Initial focus on Aspergillus . Cannabinoids in Consumables

• Initial focus on cannabinoids in hemp plant materials. • Recommendation on reporting total THC (THC + THCA ) • Recommendation on dry weight. Chemical Contaminants in Cannabis • Review Target Limit of Quantitation for pesticides. • Residual Solvents in Cannabis 5

2019 CASP Work Plan  (as of Sept. 7, 2019) 

Click  Here For Work Plan

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Advisory Panel

• composed of CASP supporting organizations • annual membership (Pioneer, Partner, Affiliate)

CASP Pioneers: ABC Testing, Inc.

PathogenDx PerkinElmer

Association of Food and Drug Officials (AFDO)

• meets by web-conference quarterly • agenda-setting & priority-ranking

Bia Diagnostics

R-Biopharm AG

Bio-Rad

SCIEX

GW Pharmaceuticals Industrial Laboratories Materia Medica Labs MilliporeSigma SōRSE Technology

Supra Research & Development TEQ Analytical Laboratories

Titan Analytical Trilogy Analytical

2019 CASP Advisory Panel – 30 Member Organizations Web-conferences: • February 14, 2019 (Midyear Meeting planning) • April 9, 2019 (workplan finalization, confirmation of working groups) • August 5, 2019 (Annual Meeting planning)

CASP Partners: CEM Corporation

Eurofins Scientific Trace Analytics

CV Sciences

CASP Affiliates: Charm Sciences Crystal Diagnostics

Lazarus Naturals Medicinal Genomics

Hygiena

SC Labs

Institute of Food Technologists (IFT)

Emerald Scientific

BIOTECON Diagnostics

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Advisory Panel – Looking Forward

Sept 2019 CASP Meeting Receive community feedback on 2019 deliverables, and future work

Oct 2019 CASP Advisory Panel Call Endorsement of DRAFT 2020 workplan

Oct – Dec 2019 Renewal & Recruitment of 2020 Advisory Panel Members Confirm membership and identify potential new stakeholders

Dec/Jan 2020 CASP Advisory Panel Call Workplan finalized based on funding commitments (potential to add deliverables/working groups) 

Mar 2020 AOAC Midyear Meeting Receive community feedback, evaluate workplan progress

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Targeting cannabis- and hemp specific standards and/or methods of analysis such as: • potency • pesticide residues • biological contaminants • chemical contaminants including select mycotoxins

• untargeted testing profiles • method validation guidelines

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Contacts Scott Coates, MSBTM CASP Program Lead Senior Director AOAC Research Institute scoates@aoac.org 301-924-7077 x137 Jonathan Goodwin, , SHRM-SCP, SPHR Deputy Executive Director & Chief HR Officer AOAC INTERNATIONAL jgoodwin@aoac.org 301-924-7077 x104 Palmer A. Orlandi, Jr., Ph.D. Deputy Executive Director and Chief Science Officer AOAC INTERNATIONAL

Christopher Dent Manager, Standards Development & Official Methods of Analysis® AOAC INTERNATIONAL cdent@aoac.org 301-924-7077 x119. Alicia Meiklejohn Governance and Business Development

AOAC INTERNATIONAL ameiklejohn@aoac.org 301-924-7077 x101

porlandi@aoac.org 301-924-7077 x163.

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2019 CASP Work Plan (as of September 7, 2019)

 Individual meetings with Advisory Panel members, experts.    Advisory Panel meeting to identify priorities.    Organize CASP Meeting in Gaithersburg, MD.    First CASP Meeting March 12 th . (A/V, catering)  

Jan to March 

 Finalize 2019 program (timetable and budget) and review with Advisory Panel.    Recruit Microbiology Working Group Chair.    Populate working groups  

April 1 to April 15 

 Convene working groups to: 1) select specific topics; 2) initial development of  SMPR; and 3) identify meetings for AOAC participation.  Three separate  conference calls.    Six conference calls per working group to develop SMPR.  Total of 18  conference calls.   Possible attendance at cannabis meetings.  FDA public hearing, IFT and AFDO. 

April 15 to April 30 

May 1 to July 15 

  Public review of draft SMPRs.        Attendance at cannabis meetings.   

July 15 to Aug 15 

  Reconcile comments and prepare final daft SMPRs.    Advisory Panel Conference Call  

Aug 15 to Sept 1 

  CASP Meeting in Denver.    Adopt 3 SMPRs.    Select 2 nd  set of topics for working group.  

Sept 7 

Advisory Panel Conference Call.  

    

Three conference calls per working group to develop SMPR.    

Oct to December 

Total of 9 conference calls.  

3 calls for methods.  

Develop initial 2020 program and confirm participation.  

Legend 

 = completed;    = in‐process ;     and   = future tasks.

AOAC Standards Development

Approval of AOAC Standards & Consensus  Documents

Deborah McKenzie Sr. Director, Standards & Official Methods SM AOAC INTERNATIONAL

ANALYTICAL AOAC Products, Services, and Analytical Excellence

Standards & Methods Development

A Complete &  Harmonized  Quality System  Through 

Official Methods of  Analysis SM (OMA) &  Performance Tested  Methods SM (PTM)

Laboratory Proficiency Testing & Quality Systems

Analytical  Excellence 

Publications, Training, Educational Outreach & Horizon‐scanning

Standards Drafting Overview

Working  Group  Meets 

• Draft 

standard

• Public 

Post draft  standard

Comment  Period        (≥ 30 days)

Reconcile  Comments 

• Recommend  final draft to  CASP

AOAC Standard Development  Process

Consensus

US National  Technology  Transfer and 

Advancement Act  (PL 104‐ 113) and  OMB Circular A‐ 119

Defensibility

Acceptability

Examples: AOAC  Consensus Products

Basic Principles

• Transparency • Openness • Balance of Interests • Due Process • Consensus • Appeals

• Performance  Requirements • Guidelines

• Sampling Standards • Methods of Analysis • Best Practices • Operational Documents

AOAC Consensus & Products

CASP Standards Development Activity

Initial CASP meeting and launch of 3 working  groups with a proposed an outline for each  working group  March 2019 – AOAC Midyear  Meeting

July ‐ August 2019

Comments on draft standard method  performance requirements Comment period for all 3 documents began  on July 2019 through August 16 2019.  Online Open Comment Session held on  August 12, 2019

Deliberate and reach consensus on a final  versions of the documents WG chairs will present summaries of draft  document for deliberation and consensus. September 7, 2019

Working groups developed draft documents:   Working groups met to begin their work and  continued drafting documents via web  conference April– July 2019

To Date – Opportunities to Participate  Balance of Perspectives & Due Process

TARGETED  COMMUNICATION

INVITATIONS TO SMES

EMAIL BLASTS &  WEBSITE  NOTIFICATIONS

PARTICIPATION IN  MEETINGS – AOAC AND  EXTERNAL MEETING

ASSOCIATION NEWS  ARTICLES

ONLINE & WRITTEN ‐ PUBLIC COMMENT  FORMATS 

BRIEFINGS & PUBLIC  HEARINGS

Global Perspectives Included

Contract Research Laboratories Independent Contractors US Rule Makers Academia Commodity Producers Product Manufacturers Instrument & Technology Providers State Regulators and Laboratories Reference Material Organizations Proficiency Testing Programs Trade Organizations Scientific Associations Rapid Method Developers Accreditation Organizations

Approving the Draft Standards

Program Lead and  Standards Manager will  oversee the approval  process

STEP 2 :   Program Lead will  conclude deliberation and  Standards Manager will record  and verify with WG chair(s) any  revisions, if needed

STEP 1: WG chair(s) introduced  to present the draft standard  along with how comments were  reconciled followed by  discussion on the draft standard

STEP 3 :  After final revisions are  complete, the WG chair will  make a motion for approval of  the standard and a second to  the motion may be entertained,  but is not necessary 

STEP 4: Program lead will  acknowledge the motion (and  second, if offered) and offer  time for discussion/questions on  the motion.  

STEP 5:   After any due  discussion, Program Lead will all  for a vote on the motion.  

STEP 6:   Standards Manager will  walk attendees through the  electronic balloting process.   All attendees will be able to  participate in the vote

NOTE: 2/3 vote in favor of a motion will  pass a motion.   2/3 of those voting will  demonstrate consensus in  passing a motion

Documentation and  Communication

AOAC carefully documents the actions  of CASP and the Working groups

AOAC will prepare summaries of the  meetings Communicate summaries to the stakeholders Publish status and summaries in the Referee section of AOAC’s  Inside Laboratory Management Publish documents in  Journal of AOAC  INTERNATIONAL, Official Methods of Analysis of  AOAC INTERNATIONAL

Roles and Responsibilities

Establish working groups to develop  standards & consensus documents Comment, deliberate, and establish voluntary  consensus standards

Stakeholder Panel

Develop draft standards & consensus  documents Reconcile comments Present draft standard to stakeholders

Stakeholder Panel Working  Groups

Official Method Board 

Provide process oversight and review

Coordinate stakeholder panel, working groups, and  facilitate their meetings Document actions/decisions of working groups and  program meetings Post draft standards, collect comments, and publish  approved standards

AOAC Staff

Questions?

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Patrick Bird & Julia Bramante Co-Chairs, CASP Microbial Contaminants Working Group Working Group Presentation

September 7, 2019 Sheraton Denver Downtown 1550 Court Place, Denver, CO, 80202

CASP Microbial Contaminants: Working Group Members AldwinAnterola, Southern Illinois University Carbondale NathanHordyk, Industrial Laboratories

MikePfaller,University of Iowa

TobyAstill,PerkinElmer

ChrisHudalla, ProVerdeLaboratories

Melissa Phillips, NIST

SusanAudino, S.A.Audino&Associates

KristenHypes,Mile HighLabs Ron Johnson,BioMerieux

AlenaRodriguez,Rm3Labs Yvonne Salfinger,AFDO Nandakumara Sarma,USP

BrianBeck,Microbiologics

Cornelius Berka,BIOTECONDiagnostics

JuanRodriguez,GreenHillsAnalytics Lab

PatBird,PMBBioTek

BenKatchman,PathogenDx

EdwardSawicki,Think20Labs Rachel Shegog,SoRSETechnology

Rafael Bombonato,Curaleaf KyleBoyar,Medicinal Genomics

JasonKircos,Neogen

JulieKowalski,TraceAnalytics Jasen Lavoie, U.S.CannabisPharma Quynh‐Nhi Le,NeogenCorporation

CaseySimmons,Hygiena

JuliaBramante,CDPHE

Sidney Sudberg,Alkemist Labs

Roger  Brauninger,A2LA (accreditation body)

SungOuiSuh,ATCC

RobertBrodnick, TitanAnalytical MikeClark,BioRad Laboratories

Raymundo Lerma,Phytatech

ChristySwoboda,Romer Labs, Inc.

Cynthia LeVesque,Caligreen Laboratory Kyle Lorenzen,Young Living EssentialOils

KatherineThomas,NJDepartmentofHealth AnandThota,Vivariant Laboratories Michael (Bhodi) Tims,Radix Medicina, LLC

BobClifford, Shimadzu

Danielle Deschene, SC Laboratories

EvaLynch,RockRiver Laboratory

LoriDodson,Maryland Medical CannabisCommission KevinMcKernan,Medicinal Genomics

TriciaVail, SartoriusCorporation

Wilfredo Dominguez,3M

MeganMurn,Microbiologics

GordonVrdoljak,StateofCADeptofPublicHealth

JanieDubois,ContractAnalytical Services

MariaNelson,AOAC

ChristopherWaggener,VirginiaDCLS

TedGatesy,MDARD

DustinNewman, Instituteof FoodSafety&Defense

MatthewWard,CDPHE

StephenGoldman,PhytaTech

GaryNiehaus,CrystalDiagnostics Melissa Nutter,TitanAnalytical

JaneWeitzel, Independent Consultant

BridgettGraham,OrganaKannalytics

DanielWene,New JerseyDepartmentofHealthPHEL

Bradford Haskell,DesignGroupCollaborative

ShawnO'Leary,NJDOH ShaunOpie,E4Bioscience BenOrsburn,Think20Labs

AnnaWilliamsWilliams,A2LA

DarinHayakawa,HawaiiDOH 

SethWong,TEQAnalytical Laboratories

YvonneHelbert,Medicinal Genomics

VictoriaWu,ABCTesting&MateriaMedica Labs

ShannonHoffman, SteepHill

JessPaoletti,TEQAnalytical Labs

JoshuaWurzer,SCLaboratories

ShermanHom,NJDOH

SalvatoreParisi,AlBalqaApplied University

ChristineYeo,NaturalProductsAssociation WendiYoung,MileHigh Labs JessaYoungblood,HardyDiagnostics

Microbial Contaminants Working Group: Work To Date

• Eight teleconferences  (April 2019 – August 2019) • One SMPR drafted • Public comment period (June – July, 2019) • SMPR made ready for review and approval 

SMPR Development

Standard Method Performance Requirements ® (SMPRs)  for  Detection of Aspergillus in Cannabis and Cannabis Products  Applicability:  Candidate methods used to detect Aspergillus  ( Aspergillus niger,  Aspergillus fumigatus, Aspergillus flavus,  and Aspergillus terreus )  in cannabis (plants/flowers) and/or cannabis products  (concentrates, infused edibles and infused non‐edibles).   Candidate methods may be validated for specific matrices,  categories or broader claims.  

Aspergillus species • A common mold that produces spores that become airborne. • The spores are easily spread through air currents and are found in most environments, both indoors and outdoors. • Most people breathe in the spores everyday without getting sick. • People with weakened immune systems are at a higher risk of developing aspergillosis. • Infections start in the lungs and can quickly lead to infection. Infections can be serious and sometimes fatal.

• Aspergillus contaminates cannabis flowers and plants. • Individuals who consume cannabis through smoking are at higher risk for infection

Definitions • POD, Probability of Detection.— The portion of positive analytical outcomes for a qualitative method for a given matrix at a given analyte level or concentration • dPOD CP – This difference in POD values between presumptive and confirmed results • LPOD – The POD value obtained from combining all valid collaborator data sets for a given matrix at a given analyte level or concentration • LCL, Lower confidence limit.— Calculated to determine 95% confidence interval of various POD values • UCL, Upper confidence limit.— Calculated to determine 95% confidence interval of various POD values • CFU, Colony forming unit.— Number of viable microorganisms, presented per a specific quantity. (ex. CFU/mL) • MPN, Most probable number.— Method used to estimate contamination levels

Method Performance Requirements

Table 1. Validation Acceptance Criteria (Plants/Flowers, Concentrates,  Infused Edibles, Infused Non‐Edibles)

Parameter  Requirements

Target Test Concentration a

Parameter

Minimum Acceptable Results

Single Laboratory Validation with artificial contamination

Low level to produce  fractional positive  results  Ex. 0.2‐2 CFU/Test  Portion High level to produce  consistently positive  results Ex. 2‐10 CFU/Test  Portion 0 CFU/Test Portion

Fractional positive results,  25‐75%   (5‐15 positive test  replicates)  dPOD CP   95% CI: LCL < 0 < UCL b

Replicates per matrix: 20 Inoculation procedure:  AOAC Appendix J 

Fractional Concentration  (low level)

Replicates: 5 Inoculation procedure:  AOAC Appendix J

High Concentration

POD of 1.00 c

Non‐Inoculated (Zero)  concentration

Replicates: 5

POD of 0.00 c

Single Laboratory Validation with natural contamination 

Fractional positive results, 25‐75% (5‐15 positive test  replicates) for minimum 1 lot dPOD CP   95% CI: LCL < 0 < UCL b

Acceptable minimum  detection level (low  level)

2 separate lots of  20 replicates

N/A

Method Performance Requirements

Table 1. Validation Acceptance Criteria (Plants/Flowers, Concentrates,  Infused Edibles, Infused Non‐Edibles)

Parameter  Requirements

Target Test Concentration a

Parameter

Minimum Acceptable Results

Multi Laboratory Validation

0.15  ≥ LPOD  ≥  0.85 

12 replicates

1‐10 CFU/Test Portion

95% CI: LCL < 0 < UCL b

dPOD CP

LPOD

12 replicates

10‐50 CFU/ Test Portion

LPOD ≥ 0.95

12 replicates

0 CFU/Test Portion

LPOD ≤ 0.05

LPOD (0)

a Determined through MPN Procedures (see Table 4) b The range between the lower and upper confidence interval should encompass 0,  if not, the results must be investigated, and an explanation provided.  c If acceptance criteria is not observed, results must be investigated, and an explanation provided. 

Method Performance Requirements

Table 6. Inclusivity/Exclusivity Performance Requirements

Final Test  Concentration  (CFU/mL)

Minimum  Acceptable Results

Parameter

Parameter Requirements

Inclusivity Single‐laboratory validation (SLV) study: At  least 10 strains per required Aspergillus spp.  (reference Annex I) cultured by the candidate  method enrichment procedure.  A minimum of  50 strains is required. Exclusivity SLV study: At least 30 non‐target organisms,  (including those required in Table 8), cultured  under optimal conditions for growth b

10‐100 x limit of  detection of the  candidate method 

100%positive results a

Overnight growth  undiluted

100%negative results a

a. 100% correct analyses are expected. All unexpected results are to be retested following internationally  recognized guidelines (ISO  16140, AOAC OMA Appendix J, The Compendium of Analytical Methods of  Health Canada). Some unexpected results may be acceptable if the unexpected results are investigated,  and acceptable explanations can be determined and communicated to method users b. In instances where an exclusivity culture produces a positive result by the candidate method, the  culture may be reanalyzed after culture following the candidate method enrichment procedure.  Both  results (optimal growth conditions and candidate methodenrichment) must be reported.

Method Performance Requirements

• Bulk inoculation of test material is required. • One matrix must contain microflora at 10x the level of the target microorganism. • MPN analysis to determine concentration of target microorganism. Use of matrix study replicates is encouraged.

• Use of live (viable) cultures and/or fungal spores (liquid stressed/non- stressed, lyophilized) is required. • To screen samples for the presence or absence of the target analyte, two methods that employ different technologies (agar plate, PCR, ELISA) must be used. • Extended primary enrichment (up to at least 48 total hours) followed by plating of the sample to a minimum of two different types of agar plates.

Method Performance Requirements

Table 2. Category Test Portion Requirements

Table 3. Acceptable Matrix Claims

Minimum Test  Portion Size a

Criteria

Category

Matrix Claim

Plants & Flowers

10 g 

Number of Matrices Minimum Number  of Categories

15 (minimum 3  matrices/category) ≥ 10 (minimum 2  matrices/category)

Concentrates

5 g

Broad Range

4 categories

Infused Edibles 

25 g

Variety

4 categories 2 categories 1 category 1 category

Infused Non‐Edibles 10 g a Minimum test portion size required for validation.   Alternatively, larger test portions may be validated.  

Select

≥ 5 ≥ 5 ≥ 1

Specific Category Specific Matrix (s)

Comments Submitted

The definition for acceptable minimum detection level does not align  with the final version presented in ISO 16140‐1. No minimum detection  level has been set as a requirement; therefore this definition has been  removed from the document.  Table 1 has been modified to emphasize  the low contamination level should produce fractional positive results.  This aligns the SMPR with Appendix J.    After spores, more descriptive language regarding the state of the  inoculum (liquid stressed/non‐stressed, lyophilized) has been added  parenthetically. The SMPR  has been updated to the following: At the time of the  publication, no national reference standard method exists for the  confirmation of Aspergillus from cannabis products.  The SMPR  has been updated to: Until a suitable reference method is  established the following is recommended for method developers. The SMPR has been updated to change probably to probable.

Line 34: The entire 90% confidence interval  on the POD falls above 0.95. 

Line 154: Add additional details regarding  condition of culture for inoculation Line 165:  Suggestion to replace word  "standard" with "method" Line 166:  Replace "until one" with "until a  suitable reference standard method ... Line 190:  "probable" not "probably"

Comments Submitted Line 190:  Add "Which if possible, should  include the replicates run in the validation  study"

See Next Comment for change made regarding Line 190

Line 191:  Right before "See appendix"  please add a new statement, if possible,  regarding the use of matrix study test  portions in the MPN study.

The following sentence has been added to the SMPR: If possible, the use  of test portions included in the matrix study  should be included as a level in the MPN study.   Contamination levels have been modified to be broader and not as  prescriptive.   AMDL has been changed to Fractional Concentration (Low Level) in  column one.   The minimum acceptance criteria has been reworded to be more  straightforward:   Fractional positive results 25‐75%  (5 to 15 positive test  replicates).  No change to the dPODCP criteria as it matches Appendix J Table 1 has been updated to indicate 12 replicates are required for each  contamination level for an MLV

Method performance table:  Review the  number of CFUs and dPOD in AMDL 

Method Performance table: Include the  number of test replicates per  contamination level for the MLV

Comments Submitted

Table 5 ‐ Conditions of inoculating culture  needs more details

Additional details have been added to describe the inoculating cells state  prior to inoculation (ex. Liquid non‐stressed) Table 6 has been updated to indicate ‐ Minimum # of strains (50) needed  to test for inclusivity;           Final test concentration of inclusivity organisms has been changed to 10‐ 100x the LOD of the candidate method.  Requirement for exclusivity cultures to be grown in non‐selective media  (and not following candidate method enrichment process) has been  added.   A footnote has been included to reference that positive exclusivity  strains may be recultured following the candidate method enrichment  procedure and analyzed a second time. Both sets of data will be  reported. All changes align the SMPR with Appendix J Use of confidence intervals for inclusivity and exclusivity testing is not  established in international validation guidelines.   ATCC strains have been kept in the table, but a footnote indicating that  the strains are recommended due to previous misidentification as one of  the four target strains.  Alternative strains for the same genus/species  may be substituted

Table 6 Inclusivity does not align with  Appendix J.

Table 6 Inclusivity:  Use of POD and 95% CI  to determine acceptance criteria

Table 8:  Remove ATCC designations

Motion: Move to accept the Standard Method Performance Requirements ® (SMPR ® ) for Detection of Aspergillus in Cannabis and Cannabis Products as presented.

Discussion?

DRAFT AOAC SMPR 2019.XXX; Version 5;  August 22, 2019  1 2 Method Name:    3

Standard Method Performance Requirements for Detection of Aspergillus  in 

4 5 6

Cannabis and Cannabis Products 

Intended Use :   7 8 1. Purpose:    AOAC  Standard Method Performance Requirements SM  (SMPRs) describe the minimum  9 recommended performance characteristics to be used during the evaluation of a method. The  10 evaluation may be an on‐site verification, a single‐laboratory validation, or a multi‐site collaborative  11 study. SMPRs are written and adopted by AOAC stakeholder panels composed of representatives from  12 industry, regulatory organizations, contract laboratories, test kit manufacturers, and academic  13 institutions. AOAC SMPRs are used by AOAC expert review panels in their evaluation of validation  14 study data for method being considered for Performance Tested Methods SM or AOAC Official Methods  15 of Analysis SM  and can be used as acceptance criteria for verification at user laboratories. [Refer to  16 Appendix F: Guidelines for Standard Method Performance Requirements , Official Methods of Analysis  17 of AOAC INTERNATIONAL  (2019) 21 st  Ed., AOAC INTERNATIONAL, Rockville, MD, USA.]  Consensus‐based Reference method.   Candidate methods used to detect  Aspergillus  ( Aspergillus niger, Aspergillus fumigatus, Aspergillus  flavus, and Aspergillus terreus)  in cannabis (plants/flowers) and/or cannabis products (concentrates,  infused edibles and infused non‐edibles). Candidate methods may be validated for specific matrices,  categories or broader claims.  See Table 3 for matrix/category claim acceptance criteria.  2. Applicability :  

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 47 48

3. Analytical Technique :

Any analytical technique that meets the method performance requirements is acceptable. 

4. Definitions :

Candidate Method . — The method submitted for validation [Appendix J:  AOAC INTERNATIONAL  Methods Committee Guidelines for Validation of Microbiological Methods for Food and Environmental  Surfaces ,  Official Methods of Analysis of AOAC INTERNATIONAL , (2019) 21 st  Ed., AOAC

INTERNATIONAL, Rockville, MD, USA] 

Candidate Method Presumptive Result . —Preliminary result for a test portion produced by following 

a candidate method’s instructions for use.

Candidate Method Confirmed Result . —Final result obtained for a test portion after cultural 

confirmation of a candidate method. 

Cannabis .—genus of flowering plants within the Cannabinaceae family that commonly contain 9‐ tetrahydrocannabinol (THC), cannabidiol (CBD), and other cannabinoids and terpenes. Cannabis 

includes, but is not limited to, high‐THC and high‐CBD cultivars. 

Cannabis Concentrates . —Extracts (primarily composed of cannabinoids and/or terpenes)  manufactured through the extraction and concentration of compounds derived from the cannabis  plant or flower.  Final products can be many forms including oils, wax, or hash (Category II). 

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

Cannabis Infused Edibles .—Food and drinks containing extracts of cannabis and/or cannabis 

materials (Category III). 

Cannabis Infused Non‐Edibles .—Products containing extracts of cannabis and/or cannabis materials  intended to be applied to the human body or any part thereof. Final products can be many forms 

including creams, ointments, cosmetics and therapeutic pads (Category IV).  

Cannabis Plant and Flower . —General terms for the structural and flowering unadulterated parts of 

the cannabis plant (Category I).  

Cannabis Products .—Products (Edible, and non‐edible) extracted or infused with compounds derived 

from the cannabis plant including but not limited to CBD and THC. 

Probability of detection (POD) .—The portion of positive analytical outcomes for a qualitative method  for a given matrix at a given analyte level or concentration. This difference in POD values between 

presumptive and confirmed results is termed dPOD CP . 

Exclusivity .—Study involving pure nontarget strains, which are potentially cross‐reactive, that shall  be not detected or enumerated by the candidate method.   See   Table 8 for a list of recommended  nontarget strains. [Appendix J: AOAC INTERNATIONAL Methods Committee Guidelines for Validation  of Microbiological Methods for Food and Environmental Surfaces ,  Official Methods of Analysis of  AOAC INTERNATIONAL , (2019) 21 st  Ed., AOAC INTERNATIONAL, Rockville, MD, USA]  Fractional positive .—Validation criterion that is satisfied when an unknown sample yields both  positive and negative responses within a set of replicate analyses. The proportion of positive  responses should fall within 25 and 75% and should ideally approximate 50% of the total number of  replicates in the set. A set of replicate analyses are those replicates analyzed by one method. Only  Inclusivity .—Study involving pure target strains that shall be detected or enumerated by the  candidate method.   See  Table 7 for a list of recommended target strains. [Appendix J:  AOAC  INTERNATIONAL Methods Committee Guidelines for Validation of Microbiological Methods for Food  and Environmental Surfaces , Official Methods of Analysis of AOAC INTERNATIONAL , (2019) 21 st  Ed.,  Laboratory probability of detection (LPOD) .—The POD value obtained from combining all valid  collaborator data sets for a method for a given matrix at a given analyte level or concentration.  [Appendix H:  Probability of Detection (POD) as a Statistical Model for the Validation of Qualitative  Methods ,  Official Methods of Analysis of AOAC INTERNATIONAL , (2019) 21 st  Ed., AOAC one set of replicates per matrix is required to satisfy this criterion.  AOAC INTERNATIONAL, Rockville, MD, USA] 

INTERNATIONAL, Rockville, MD, USA] 

LCL .—Lower confidence limit. 

Aspergillus .—Filamentous, cosmopolitan and ubiquitous fungus found in nature producing colonies  typically of 1‐9 cm in size (select species produce 0.5‐1 cm colonies). Colonies are powdery in texture  and color varies based on species.  Reverse color is typically uncolored to pale yellow. Growth is typical 

96 97 98 99

at 20‐30 o C. Aspergillus fumigatus  is thermotolerant and can grow at a temperature range of 20 to 50  °C. For all species, hyphae are septate and hyaline. The conidiophores originate from the basal foot  cell located on the supporting hyphae and terminate in a vesicle at the apex. Vesicle is the typical  formation for the genus  Aspergillus . The morphology and color of the conidiophore vary from one  species to another. Covering the surface of the vesicle entirely (“radiate” head) or partially only at the  upper surface (“columnar” head) are the flask‐shaped phialides which are either uniseriate and  attached to the vesicle directly or are biseriate and attached to the vesicle via a supporting cell,  metula. Over the phialides are the round conidia (2‐5 µm in diameter) forming radial chains. Other  microscopic structures include sclerotia, cleistothecia, aleuriconidia, and Hulle cells are of key  importance in identification of some Aspergillus  species. Cleistothecium is a round, closed structure  enclosing the asci which carry the ascospores. The asci are spread to the surrounding when the  cleistothecium bursts. Cleistothecium is produced during the sexual reproduction stage of some  Aspergillus species. Aleuriconidium is a type of conidium produced by lysis of the cell that supports it.  The base is usually truncate and carries remnants of the lysed supporting cell. These remnants form  annular frills at its base. Hulle cell is a large sterile cell bearing a small lumen. Similar to cleistothecium,  it is associated with the sexual stage of some  Aspergillus  species. See Table 9 & 10 for more 

100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141

macroscopic and microscopic information on Aspergillus  species. 

Chen, S.C.A., Meyer, W., Sorrell, T.C. , Halliday, C. L. (2019) in Manual of Clinical Microbiology ,  12th Ed, Landry, M.L., McAdam, A.J., Patel, R., & Richter, S.S. (Eds)ASM Press, Washington, D. C., , 

pp. 2103‐2131. 

Anaissie, E.J., McGinnis, M.R., Pfaller, M.A. (2009) in Clinical Mycology , Ed 2, Churchill 

Livingstone, New York, NY, pp 1‐687. 

Walsh, T.J., Hayden, R.T., Larone, D.H. (2018) in  Larones Medically Important Fungi: A Guide to 

Identification , 6 th  Ed, ASM Press, Washington, D.C., pp. 1‐500 

Test portion . —The test portion is the sample size used in most validation studies. For cannabis  flower/plant and cannabis infused non‐edible products a 10 g test portion is used.  For cannabis  concentrates, a 5 g test portion is used. For cannabis infused edibles, a 25 g test portion is used.  A  larger test portion can be used in validation studies when appropriate. See Table 2 for minimum test 

portion requirements. 

United States Pharmacopeia. Microbiological Examination of Nonsterile Products: Microbial 

Enumeration Tests (61), USP 40. United States Pharmacopeia. 

United States Pharmacopeia. Microbiological Examination of Nonsterile Products: Tests for 

Specified Microorganisms (62), USP 40. United States Pharmacopeia. 

Feng, P.,  Weagant, S.D.,  Grant, M.A.,  Burkhardt, W. (2017) Bacteriological Analytical Manual: 

Chapter 4 Enumeration of Escherichia coli and the Coliform 

Bacteria https://www.fda.gov/Food/FoodScienceResearch/LaboratoryMethods/ucm064948.htm Andrews, W. H.,  Wang, H., Jacobson, A.,  Hammack, T. (2018) Bacteriological Analytical Manual  

Chapter 5: Salmonella

https://www.fda.gov/Food/FoodScienceResearch/LaboratoryMethods/ucm070149.htm

UCL . —Upper confidence limit. 

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