CASP Meeting Book AOAC Annual Meeting 2023

AOAC INTERNATIONAL Cannabis Analytical Science Program AOAC INTERNATIONAL 2023 Annual Meeting Tuesday, August 29, 2023 1:00 – 3:00pm

Meeting Agenda I. WELCOME, INTRODUCTIONS AND ANNOUNCEMENTS (1:00PM – 1:05PM) Allison Baker (A OAC INTERNATIONAL ) will provide an overview of recent CASP highlights and a summation of recent Advisory Panel meetings. II. AOAC LABORATORY PROFICIENCY TESTING PROGRAM (1:05PM– 1:30PM) Shane Flynn ( Sr. Director, Laboratory Proficiency Testing Program, AOAC INTERNATIONAL ) will give an overview and update on the Cannabis and Hemp Proficiency Testing Program • UPDATES ON FIRST CANNABIS TESTING ROUND Danielle Lenoir ( Signature Science ) will provide updates on the first cannabis testing round results. • PROFICIENCY TESTING DEVELOPMENT – MICROBIOLOGICAL IMPURITIES Danielle Lenoir ( Signature Science ) will provide an overview of the microbiological impurities PT program currently in development. III. GUIDANCE FOR DETERMINING MOISTURE CONTENT (1:30PM – 1:50PM) Conner Jeffries (ADDIUM, Inc.) and Jini Glaros (ModernCanna) will present information on moisture content determinations in cannabis and provide a historical precedent for using dry weight basis. IV. THE IMPORTANCE OF SAMPLING (1:50PM – 2:15PM) Polona K. Carson, Ph.D., MBA, (Neptune and Company, Inc.) will present on best practices for sampling of heterogeneous materials to the cannabis community.

V. PESTICIDES IN CANNABIS MATRICES (2:15PM – 2:55PM)

• COMMNUITY METHOD DEVELOPMENT ‘THINK TANK’

Julie Kowalski (Kowalski Science Support) will provide an overview of AOAC SMPR 2018.011, Identification and Quantitation of Selected Pesticide Residues in Dried Cannabis Materials, review challenges of the SMPR, and discuss the benefits of a community method development initiative . • RECOMMENDED PESTICIDE LOQs FROM ACIL

Joe Konschnik (Plus One Consulting LLC) will highlight ACIL’s work and basis for the recommended LOQs for pesticides in the GUIDE TO HARMONIZING CANNABIS LABORATORY QUALITYAND TESTING PRACTICES Guidance Document.

VI. WRAP UP AND ADJOURN (2:55PM – 3:00PM)

This agenda can change without notice.

Allison Baker, AOAC INTERNATIONAL Allison works as a Coordinator of Standards and Official Methods, suppor�ng standards development ac�vi�es at AOAC INTERNATIONAL. She is the manager for the Cannabis Analy�cal Science Program (CASP), the Analy�cal Interna�onal Methods and Standards (AIMS) Program, and coordinates addi�onal working group ini�a�ves. Prior to joining AOAC, Allison worked at the Frederick Na�onal Laboratory for Cancer Research as a research associate for the Center for Molecular Microscopy, and at Leidos, serving as a scien�fic manager for the

Department of Defense’s Congressionally Directed Medical Research Programs. Allison graduated from Mount Saint Mary’s University in 2017 with a B.S in health sciences and again in 2019 with a M.Sc. in biotechnology and business management.

Shane Flynn, AOAC INTERNATIONAL Shane Flynn is the Senior Director of the AOAC Laboratory Proficiency Tes�ng (PT) Program. Shane helped start the first accredited Proficiency Tes�ng program for food labs in the United States 24 years ago at AOAC and has been developing new programs to address the needs of the analy�cal laboratory community since. He is currently working on Cannabis and Hemp PT programs. In addi�on to his Proficiency Tes�ng role, he also is the staff lead for the Analy�cal Laboratory Accredita�on Criteria Commitee (ALACC) and the Technical Division of Laboratory Management (TDLM). Danielle Lenoir, Signature Science Danielle Lenoir is a Senior Quality Assurance Chemist and Program Manager for the Hemp and Cannabis proficiency tes�ng program at Signature Science, in conjunc�on with AOAC INTERNATIONAL. Her focus is on developing and tailoring tools to help laboratories establish and maintain regulatory compliance, as well as developing solu�ons for complicated GC obstacles. She has over 10 years of experience in quality assurance, gas chromatography, and method development working with oil & gas, environmental, and government laboratories.

Joe Konschnik, Plus On e Consul�ng, LLC.

As Principal Consultant for Plus One Consul�ng LLC Joe applies his 40 years of analy�cal chemistry knowledge and experience to support his clients in the analy�cal tes�ng industry. Prior to re�ring from RESTEK in January, 2023 Joe was RESTEK’s Business Development Manager for the global Food & Agriculture market where he iden�fied market needs, new technologies, new product opportuni�es and coordinated technical content development for new workflow solu�ons. Prior to RESTEK Joe spent 19 years working for some of the na�on’s largest environmental tes�ng laboratory companies as a bench chemist and in

technical and laboratory opera�ons senior management roles. Joe has been an invited speaker on sampling and analy�cal chemistry topics globally. He serves as a volunteer in mul�ple AOAC working groups and communi�es and as a member of the TDRM Execu�ve Commitee. Joe has also previously served in volunteer leadership roles for the Independent Laboratories Ins�tute (ILI), the American Council of Independent Labs (ACIL)’s Food Sciences Sec�on and The North American Chemical Residue Workshop (NACRW). Direct correspondence at joe@plusone.llc, or joe.konschnik@restek.com.

Julie Kowalski, JA Kowalski Science Support LLC Julie Kowalski is a technical consultant serving primarily the cannabis, hemp and food tes�ng markets. She partners with clients offering technical exper�se and advise taking advantage of her over 20 years of laboratory experience. She specializes in developing fit -for- purpose technical programs and customized training for laboratory staff. Julie has a passion for analy�cal chemistry, training and the cannabis tes�ng industry, star�ng work on cannabis tes�ng in 2011. Julie is a sought - a�er speaker and trainer focusing on educa�onal presenta�ons on various aspects of analy�cal chemistry.

Julie earned her graduate degree in Analy�cal Chemistry from Pennsylvania State University. Her professional experience includes troubleshoo�ng, method development and valida�on for GC, GC - MS, LC, and LC - MS/MS with exper�se in pes�cide residue analysis and chromatography method development. She worked at a technology provider for over ten years developing methods and products including designing reference materials for specific applica�ons. Recently Julie served as Scien�fic Director and Chief Scien�fic Officer for a cannabis tes�ng lab. She has previously served as the President and Program chair of the North American Chemical Residue Workshop, served on AOAC Expert Review Panels, the Cannabis Scien�fic Task Force for Washington State and is currently co - chairing the AOAC CASP Chemistry Working Group. She is a member of the CASP PT and Training working groups. Julie is also a member of the AOAC CASP Proficiency Tes�ng Task Force. In addi�on to technical skills, Julie is an a uditor for an accredita�on body working primarily with laboratories seeking accredita�on to ISO 17025.

Conner Jeffries, ADDIUM Food R&D Lab Conner Jeffries is a scien�st in the ADDIUM Food R&D Lab. He earned his B.S. in chemistry from the University of Washington in 2008, and completed his M.S. in chemistry from the University of Idaho in 2013 researching synthe�c and materials chemistry. He pursued doctoral research in chemistry at the University of Idaho un�l 2017, at which point he le� academia for industry. His work in the Food R&D Lab helps ADDIUM understand the interac�ons between water molecules, food, and the AQUALAB instruments we develop. Jini Glaros, Modern Canna Labs Jini Glaros is the Chief Scientific Officer (CSO) at Modern Canna Labs in Lakeland, FL. She received her bachelor’s degree with honors in Biochemistry from the University of West Florida. In May of 2022, Jini earned a master’s degree in Medical Cannabis Science and Therapeutics from the University of Maryland, Baltimore. Before entering the cannabis industry, Jini gained valuable research experience at several well -

respected institutes, including Emory University, the National Institute of Aging, and the University of Florida. She became the laboratory director of Modern Canna in January of 2018 and was promoted to CSO in August of 2022. Jini is extremely involved in the cannabis industry and currently serves on the Scientific Advisory Board for Analytical Cannabis and CloudLIMS, is a member of the CASP Advisory Panel and the Proficiency Testing Advisory Task Force with AOAC International, is a member of the Cannabis Working Group (D37) with ASTM International, and was a member of the CannMed 2023 Advisory Board.

Polona Carson, Neptune and Company, Inc. Polona Carson, PhD, MBA has more than 30 years of experience working on multidisciplinary teams in the area of quality assurance, environmental science, analytical chemistry, materials science, and management. She started her career at Slovenian Forestry Institute where she managed the Laboratory for Forest Ecology and studied sampling of plant materials and soil. Her dissertation was on optimization of soil sampling designs in a long - term forest monitoring

program. She is currently working as a consultant and project manager at Neptune and Company, Inc., where she is utilizing her strong analytical skills, statistical knowledge, and her previous analytical laboratory management experience to provide support to laboratories and reference materials producers. She is an active member of ASTM Committee E11 Quality and Statistics.

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 | New Orleans, LA •

AOAC INTERNATIONAL CANNABIS ANALYTICAL SCIENCE PROGRAM

(CASP) MEETING Tuesday, August 29, 2023 1:00PM – 3:00PM CDT

Please scan to sign into the meeting and gain access to meeting materials!

Support for current scope in CASP is made possible by the following:

PIONEER Deibel BioScience PARTNER Modern Canna

Eurofins Scientific

MilliporeSigma AFFILIATE SPEX an Antylia Scientific Company Association of Food and Drug Officials

Medicinal Genomics

ADDIUM PSI Labs Restek

HORIBA Scientific

Hygiena

R-Biopharm Rhone GOVERNMENT MD Department of Agriculture

NV Cannabis Compliance Board IL Department of Agriculture

CO Department of Public Health & Environment

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

Today’s Agenda  CASP and PT Updates  Cannabis Dry Weight  Sampling of Heterogeneous Materials  Method Development “Think Tank” – Pesticides  Pesticide LOQs in Cannabis

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA CASP Updates and Progress • Webinars, Speaking Opportunities, and Collaborations • CANNRA Standards Webinar with AOAC, USP, and ASTM • CANNRA External Stakeholder Panel (June 2023) • AOAC CASP/CANNRA Method Validation Training Course (August 2023) • AOAC Cannabis Analytical Science Program Session at Cannabis Science Conference Fall • General Working Group Updates • CASP Microbial Contaminants WG • Work on SMPR for Detection and Enumeration of Listeria monocytogenes in Cannabis Infused Edibles • Comment reconciliation for Micro Validation Guidelines • CASP Chemistry Working Group • CALL FOR METHODS: Beverages in Cannabis – Coming Soon! • Method Development “Think Tank” – Pesticides

CASP Deliverables

Completed Deliverables

AOAC CASP Priority Subtopics

Performance Tested Methods

Official Methods of Analysis

Proficiency Testing

CASP was formed to provide consensus-driven standards and methods to promote accuracy in label potency claims and to address public safety issues Standard Method Performance Requirements Detailed description of requirements and parameters for candidate methods CASP Consensus-Based Methods Aim to be accurate and repeatable

SMPRs

Aspergillus E. Coli (STEC) Salmonella













Microbiology







Listeria



Yeast and Mold









Pesticides





Residual Solvents

Chemical Contaminants



Mycotoxins Heavy Metals











Cannabinoids

Hemp (Low THC) Cannabis > 0.3% THC













Terpenes



AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

Applicable Matrices Covered in CASP-Deliverables

Applicable Matrices Covered in AOAC Completed Deliverables Microbiology Chemical Contaminants Cannabinoids/Terpenes

Cannabis and Cannabis Product

Cannabis Dried Plant Material

Concentrates

Dried Cannabis and Hemp Flower Cannabis Concentrates

Extracts

THC-Infused Chocolate

Cannabis and Cannabis Product

Dried Plant Material Edible Chocolates Infused Beverages Cannabis Flower

Infused Beverages

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

AOAC Official Methods of Analysis (OMA)

Applicable SMPR(s)

Analyte(s)

OMA Number

Method Name

Matrix(es)

997.02 3M™ Petrifilm™ Yeast and Mold Count (YM) Plates Foods and dried cannabis flower

Yeast and Mold

2021.009

Enumeration of Yeast and Mold in Foods, Dried Cannabis Flower, and Selected Surfaces

2014.05

Foods and dried cannabis flower

2017.001 2017.002 2017.001 2017.002 2022.001

2018.10

Cannabinoids in Dried Flowers and Oil

Dried cannabis flowers and oils

Cannabinoids

Dried cannabis and hemp materials, concentrates, and oils

2018.11 Quantitation of Cannabinoids in Cannabis Dried Plant Materials, Concentrates, and Oils

Dried plant materials, concentrates, oils, extracts, tinctures of cannabis, and cannabis-related products.

Heavy metals

2021.03 Heavy Metals in a Variety of Cannabis and Cannabis-Derived Products

2020.001

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

CASP Guidance Documents:

Laboratory Guidance - Drying Field-Fresh Hemp Plant Samples in Preparation for Determination of Total THC on a Dry-Weight Basis Provides general guidance for laboratories to aid in the development of SOPs appropriate for processing hemp plant materials received fresh from the field.

Cannabis Appendix for the AOAC Accreditation Guidelines for Laboratories (ALACC) An Aid to Interpretation of ISO/IEC 17025:2017 created by the Analytical Laboratory Accreditation Criteria Committee (ALACC) to provide a section-by-section interpretation of requirements. (Revision coming soon!)

Guidelines for Validation of Microbiological Methods for Cannabis and Cannabis Products Provides comprehensive AOAC technical guidelines for conducting microbiological validation studies for analysis methods of cannabis and

cannabis products (Not yet finalized)

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

Get Involved in CASP

Link to Interest Form

Link to WG Sign Up

Learn more about the CASP Advisory Panel

Join a CASP Working Group!

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

Shane Flynn, Sr. Director, AOAC PT Program

Shane Flynn is the Senior Director of the AOAC Laboratory Proficiency Testing (PT) Program. Shane helped start the first accredited Proficiency Testing program for food labs in the United States 24 years ago at AOAC and has been developing new programs to address the needs of the analytical laboratory community since. He is currently working on Cannabis and Hemp PT programs. In addition to his Proficiency Testing role, he also is the staff lead for the Analytical Laboratory Accreditation Criteria Committee (ALACC) and the Technical Division of Laboratory Management (TDLM).

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

Proficiency Testing

Shane Flynn Senior Director

AOAC INTERNATIONAL Midyear Meeting March 13 –17, 2023 | Gaithersburg, MD

Cannabis/Hemp PT

• Selecting a PT Program • What sets AOAC PT apart • Q4 Pilot Rounds • TDLM

AOAC INTERNATIONAL Midyear Meeting March 13 –17, 2023 | Gaithersburg, MD

Selecting a PT Program

• Fit for Purpose

• Matrix

o Like routine samples

– Spiked by participating lab – Lyophilized pellets for micro

• Risk

• Statistics

• Overall Impact on Quality System

o Determination of assigned value – Consensus – Reference lab values – Spiked value of test material lab – others

AOAC INTERNATIONAL Midyear Meeting March 13 –17, 2023 | Gaithersburg, MD

What Sets AOAC PT Apart

• In addition to Hemp, AOAC can ship >0.3% delta-9-THC Cannabis flower, edibles, oils. • AOAC samples arrive homogeneous and ready to analyze, no spiking required • Use of reference labs for statistics (most competitors use consensus) • Scientific Association with many SMEs • Developed through CASP based on feedback from over 500 stakeholders • Less expensive than top competitors

AOAC INTERNATIONAL Midyear Meeting March 13 –17, 2023 | Gaithersburg, MD

Q4 New Program Pilots

• Micro Contaminants – Aspergillus

• Edibles (<0.3% THC)

• Adding Residual Solvents • >0.3% THC in 2024

– E. coli (STEC) – Salmonella – Listeria

• Oils (<0.3% THC)

– TAC – Y&M – CPS – Coliform – BTGN • PT Advisory Task Force

• Adding Residual Solvents • >0.3% THC in 2024

• Foreign Matter

AOAC INTERNATIONAL Midyear Meeting March 13 –17, 2023 | Gaithersburg, MD

Test Material Provider

AOAC INTERNATIONAL Midyear Meeting March 13 –17, 2023 | Gaithersburg, MD

Danielle Lenoir, Signature Science

Danielle Lenoir is a Senior Quality Assurance Chemist and Program Manager for the Hemp and Cannabis proficiency testing program at Signature Science, in conjunction with AOAC INTERNATIONAL. Her focus is on developing and tailoring tools to help laboratories establish and maintain regulatory compliance, as well as developing solutions for complicated GC obstacles. She has over 10 years of experience in quality assurance, gas chromatography, and method development working with oil & gas, environmental, and government laboratories.

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

AOAC INTERNATIONAL Cannabis/Hemp Proficiency Test Program

Danielle M. Lenoir, MS, PMP, CQA Signature Science, LLC Past, Present, and Future

Program Overview  Created based off industry feedback (CASP)

 Designed & Implemented by AOAC and SigSci

 ISO 17043 Accreditation (scope addition under review)

 First program to offer a true cannabis matrix (>0.3%)

Main Objective: Deliver PTs that provide meaningful performance information to laboratories, regulators, and other stakeholders

Page 19

Historical PT Challenges in Hemp/Cannabis Testing  Not representative  Lack of a true matrix  Homogeneity and stability  Heterogeneous samples can affect results evaluation  Not ready to analyze  Requires spiking  Unrealistic contaminant application

Page 20

Past: PT Rounds Thus Far… Pilot Round Hemp Flower May 2022 Cannabinoids Terpenes Heavy Metals

 Participants can select sample type of interest (naturally occurring vs contaminants)  Labs have ~3 weeks to analyze and report  Labs may use any method  Validated in-house, AOAC, etc.  Results evaluated statistically using reference lab and SigSci homogeneity/stability data

Moisture Pesticides

Live Round Hemp Flower

March 2023

Cannabinoids Terpenes Heavy Metals Moisture Water Activity Pesticides Mycotoxins Cannabinoids Terpenes Heavy Metals Moisture Water Activity

Pilot Round Cannabis Flower

June 2023

Page 21

Selected Results

Page 22

Terpene Results

Evaluated 9 terpenes Reference labs vs. Participants  Differing degrees of variability (may be due to methodology differences?)

α - Pinene

α - Bisabolol

α - Humulene

β - Myrcene

β - Pinene

Limonene Linalool

Terpineol

Guaiol

Page 23

Pesticide Results

Azoxystrobin

Carbofuran

Dimethoate

Malathion

Metalaxyl

Thiamethoxam

Page 24

Moisture Results

Page 25

Moisture Results

P total THC(dry) = P total THC(wet) /[1-(P moisture/100)]

Page 26

Moisture Results

P total THC(dry) = P total THC(wet) /[1-(P moisture/100)]

Page 27

Page 28

Present  Current sample design available with additional analytes covered  Currently working to increase accumulated heavy metals present in matrix material

 Different routes of contamination and cultivation conditions

Page 29

Reports  Updated

reports based on feedback from regulators and participants

 “Acceptable”

determination

Page 30

Feedback from Participants

“When analysts saw that the samples were PTs, some of them struggled to understand the fact that they could process the samples without any modifications.”

“Participating in the AOAC Cannabis/Hemp PT Program was a refreshing experience.”

“Real PT materials and samples are a game changer for the cannabis industry.”

“Thank you for making this process so simple.”

Page 31

Coming Soon…  Expanded chemistry matrices  Edibles  Oils Pilot: Late 2023 Live Round: 2024

Edibles

Cannabinoids Heavy Metals Water Activity Pesticides Mycotoxins Cannabinoids Terpenes Heavy Metals Water Activity Pesticides Mycotoxins

Oils

 Additional methods  Residual solvents  Foreign material (separate sample)

In Progress

Page 32

 Microbiological Impurities PT Scheme  2023/2024 Pilot  Includes comprehensive qualitative and quantitative panels  2-week analysis period Also in Progress…

Page 33

Example Microbial PT Schemes

Example Quantitative Duplicate Sample Inoculation Scheme Method Sample 5 Sample 6 Sample 7

Sample 8

TAC-Total Aerobic Count

-

-





CPS-Coagulase Pos Staph (S. aureus) TYM Total Yeast & Mold BTGN BileTolerant Gram Negative (e.g. Enterobacteriaeae)

- -

- -

Example Qualitative Sample Inoculation Scheme Sample 1 Sample 2 Sample 3 Sample 4









Salmonella E. coli STEC L. mono A. niger

- -

- -





 Qualitative

Total Coliform Count





 Salmonella  E. coli (STEC)  Aspergillus  Listeria

 Quantitative

 Total Aerobic Count (TAC)  Coagulase Pos Staph (S.aureus) (CPS)  Total Yeast & Mold (TYM)  BileTolerant Gram Negative (BTGN)  Total Coliform Count

Page 34

Danielle M. Lenoir, MS, PMP, CQA Hemp/Cannabis PT Program Manager Thank You!

Signature Science, LLC dlenoir@signaturescience.com (512) 533-2063 CHPT@signaturescience.com

AOAC INTERNATIONAL Cannabis_PT@AOAC.org

Page 35

Conner Jeffries, ADDIUM Food R&D Lab

Conner Jeffries is a scientist in the ADDIUM Food R&D Lab. He earned his B.S. in chemistry from the University of Washington in 2008, and completed his M.S. in chemistry from the University of Idaho in 2013 researching synthetic and materials chemistry. He pursued doctoral research in chemistry at the University of Idaho until 2017, at which point he left academia for industry. His work in the Food R&D Lab helps ADDIUM understand the interactions between water molecules, food, and the AQUALAB instruments we develop.

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

Jini Glaros, Modern Canna Labs

Jini Glaros is the Chief Scientific Officer (CSO) at Modern Canna Labs in Lakeland, FL. She received her bachelor’s degree with honors in Biochemistry from the University of West Florida. In May of 2022, Jini earned a master’s degree in Medical Cannabis Science and Therapeutics from the University of Maryland, Baltimore. Before entering the cannabis industry, Jini gained valuable research experience at several well-respected institutes, including Emory University, the National Institute of Aging, and the University of Florida. She became the laboratory director of Modern Canna in January of 2018 and was promoted to CSO in August of 2022. Jini is extremely involved in the cannabis industry and currently serves on the Scientific Advisory Board for Analytical Cannabis and CloudLIMS, is a member of the CASP Advisory Panel and the Proficiency Testing Advisory Task Force with AOAC International, is a member of the Cannabis Working Group (D37) with ASTM International, and was a member of the CannMed 2023 Advisory Board.

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

29 08 2023

Cannabis Dry Weight Moisture Content & Potency

Conner Jeffries, M.S. R&D Food Lab, Addium Jini Glaros, M.S. CSO, Modern Canna Labs

29 08 2023 Cannabis Dry Weight - Moisture Content & Beyond Presentation Goals

40

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

● Provide background and guidance on the implementation of dry weight in cannabis analysis, its effect on potency, and role in lab shopping . ● Help CASP take a stance on dry weight. ● Discuss the need for a best practice document, and potentially convene a working group in order to produce said document.

Presen tation Goals

41

29 08 2023 Cannabis Dry Weight - Moisture Content & Beyond What is dry weight and how do we determine it?

42

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

● Hemp and cannabis are legally defined in the 2018 Farm Bill by their dry weight THC content (0.3%). ○ This definition goes back to the 1970s, but that’s another story… ● 2021 Final Rule: “Dry weight is a basis for expressing the percentage of a chemical in a substance after removing the moisture from the substance.” 1 ● No definition in Farm Bill for moisture . OED: Water or other liquid diffused through a solid. 1) https://www.federalregister.gov/documents/2021/01/19/2021-00967/establishment-of-a-domestic-hemp-production-program

Context

43

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

● Technical and literature definitions of moisture indicate that moisture content is water content (FDA, USFS, NOAA, IFT, EPA, etc). 2 ● Wet weight - water content = dry weight ○ Wet basis: wet weight is 100% ○ Dry basis: dry weight is 100%

Dry Weight

2)https://www.fda.gov/inspections-compliance-enforcement-and-criminal-investigations/inspection-technical-guides/water-activity-aw-foods ; https://www.fs.usda.gov/research/treesearch/37428; https://www.weather.gov/lmk/humidity ; https://www.ift.org/news-and-publications/food-technology- magazine/issues/2009/november/columns/laboratory ; Park, Y.W. (1996) Determination of moisture and ash contents of foods in Handbook of Food Analysis, Leo, M.L.N. (Ed), Marcel Dekker, New York. pp. 59–92 ; Makower, B. (1950) Determination of water in some dehydrated foods in Analytical Methods in the Food Industry, Advances in Chemistry Series 3, American Chemical Society, Washington

44

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

Dry Weight 2 g glucose + 2 g fructose in 100 g of solution 2/100 = 2% glucose on a wet basis 2/4 = 50% glucose on a dry basis.

45

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

● Dry weight and dry basis are often used in cases where water is added or removed naturally, or is a common processing step. ○ Milk powder ○

Dry Weight

Grain ○ Wood ○ Soil ● These matrices share: ○ Thermal stability ○

Low concentrations of other volatile compounds

46

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

47 ● Use dry weight basis when water content is in flux . Example: A substance dries out over time. Using wet basis , the concentration of an analyte will increase over time. This can be problematic with upper thresholds (i.e. hemp becomes cannabis if THC > 0.3% ). Dry weight basis makes a lot of sense for dried cannabis flower, right?

Dry Weight

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

● To measure moisture content, we have to separate water from the matrix . ○ Vaporization (heat) ● Loss on drying ● Desiccation ○ Extraction (solvent) ● Distillation ● Titration

Well…

48

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

Well…

Loss on drying ● This works when we can safely assume mass lost on heating is only water . ● Cannabis flower has low thermal stability and a high concentration of other volatile compounds .

49

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

At 103 to 105 ºC , cannabinoids are decarboxylated. Jini Glaros, Modern Canna Labs.

50

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

As-is sample

Vacuum desiccation dried sample

Moisture balance dried sample

Terpenes are lost during high temperature loss on drying. Jini Glaros, Modern Canna Labs.

51

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

VOC peak

H 2 O peak

Combustion H 2 O

VOC & moisture comparison of 80 ºC & 105 ºC heating of cannabis samples. Jeff Gast, LECO, 2020.

52

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

Solvent Methods

Well…

…have their own issues

● Karl Fischer Titration ○ High cost ○

Wet chemistry pipeline ○ Plant material requires oven attachment ○ High temperatures can cause condensation and decomposition reactions. 3 ● Distillation - AOAC Method 945.21 (1945) ○ Requires a large sample size (~10 g) ○ Long process 3) Kestens, V., Conneely, P., Bernreuther, A. (2008) Food Chem. 106 , 1454-1459. doi:10.1016/j.foodchem.2007.01.079

53

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

Karl Fischer oven determination of water content in tobacco samples. V-KFT is reference extraction method. 3) Aydin, N., Chardonnens, F., Rotach, M. (2012) Contrib. To Tobacco Research. 25 , 350-359. doi: 10.2478/cttr-2013-0914

54

29 08 2023 Cannabis Dry Weight - Moisture Content & Beyond What is the current analytical landscape?

55

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

● AOAC pilot PT data (2022) ● NIST data (2022) ● Internal data (2022)

Best Moisture Content Method?

No standard method

56

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

57

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

NIST Cannabis Laboratory Quality Assurance Program: Exercise 2 Moisture Final Report. Walter Wilson, et al, November 2022.

58

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

Addium internal cannabis moisture content method comparison, 2022.

60

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

61

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

Effect of Moisture Content on Potency

Three different methods give 5%, 7%, and 15% moisture content. How does this change %THC?

20% as-is THC content

5% = 95% dry weight 7% = 93% dry weight 15% = 85% dry weight

20% 0.93

20%

20% 0.95

= 21.05%

= 21.51%

= 23.53%

0.85 16.2% relative percent difference between as-is and 85% dry weight.

62

29 08 2023 Cannabis Dry Weight - Moisture Content & Beyond Benefits, risks, & alternatives to using dry weight

63

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

Benefits , risks, & alternati ves to using

Benefits ● Water content is relevant to producers who want to maximize product yield and maximize extraction efficiency . ● Accuracy of both dosage and contaminants relies on dry weight. ● No concentration creep as volatiles are lost - i.e. when using wet-basis in hemp.

64

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

Benefits , risks, & alternati ves to using

Risks ● High heat methods can cause chemistry to happen, which inflates the moisture content. ● Without standard methods, laboratories are free to use methods that give a wide range of THC values.

65

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

Benefits, risks, & alternatives to using dry weight

Alternatives ● Use as-is weight for THC concentration (potency) ○ Removes conflicting interest for labs - no longer able to inflate market numbers.

66

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

Water Activity

67

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

● Moisture content is inherently a “path based” measure: work has to be done to separate water. ● Water activity ( a w ) is state based, and independent of mass. ○ a w is important for cannabis quality, but a w alone is not enough to know concentration data . ● A moisture sorption isotherm, describes the relationship between a w and water content ○ Requires a known moisture content to pin the data. ○ Much like spectroscopic methods.

Water Activity

68

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

Water Activit y

69

Cannabis Dry Weight - Moisture Content & Beyond Closing Thoughts & Discussion

29 08 2023

70

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

● What methods would we suggest? ● Does the USDA need to define moisture ? Is moisture specific to water? ● USDA 2021 Final Rule on domestic hemp : “A comment noted the importance of moisture content consistency in compliance sampling ... They also noted the need for best practices to be identified for drying sample material.”

Closin g Thoug hts

72

AQUALAB

Cannabis Dry Weight - Moisture Content & Beyond

29 08 2023

● It is imperative that regulators and laboratories adopt a standardized method to determine dry weight in cannabis flower. ● In hemp, does it make sense to remove all volatiles to determine dry weight? ● Remove lab shopping incentives for dry weight in hemp & cannabis. ● Should the USDA better define dry weight in the 2023 Farm Bill?

Discussion

73

DD MM YYYY

Q&A

Thank You!

Polona Carson, Neptune and Company, Inc.

Polona Carson, PhD, MBA has more than 30 years of experience working on multidisciplinary teams in the area of quality assurance, environmental science, analytical chemistry, materials science, and management. She started her career at Slovenian Forestry Institute where she managed the Laboratory for Forest Ecology and studied sampling of plant materials and soil. Her dissertation was on optimization of soil sampling designs in a long-term forest monitoring program. She is currently working as a consultant and project manager at Neptune and Company, Inc., where she is utilizing her strong analytical skills, statistical knowledge, and her previous analytical laboratory management experience to provide support to laboratories and reference materials producers. She is an active member of ASTM Committee E11 Quality and Statistics.

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

Sampling of Heterogenous Materials Polona Carson, MBA, PhD pcarson@neptuneinc.org

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

76

Why is sampling important?

Samples taken represent the product sampled All data used for characterizing the product are based on testing

Product release decision Information on the label Testing results are only as good as a sample tested

Sampling and decisions

Decision unit

Bulk materia l

Physical sample

Sampling

Inference

Lab sample

Sample

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

78

Sampling steps

Single sample of multiple increments for compositing

Sampling purpose

Primary sample

Crushing and/or splitting, drying

Physical preparation (sieving, grinding, splitting, homogenization,….)

Laboratory sample

Test sample

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

79

Heterogeneity

Materials consist of different elements: Compositional heterogeneity Distributional heterogeneity

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

80

Sampling heterogenous materials

Need to capture all compositional and all distributional elements of the material Controlling mass and number of increments taken Homogenization

© 2013 P&J Carson Consulting, LLC

81

Sampling plan

Sampling purpose Identify material to be sampled Identify measured properties Range

Resolution Sensitivity Variability Sample size and design When, how much, how? Equipment Preservation, packaging, transportation Documentation Roles and responsibilities Chain of custody! Safety Quality

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

82

Sample quality

Depends on: Previous knowledge of the material

Planning effort Sampler’s skill

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

83

Sampling purpose and measurement

What is the reason to take a sample? Characterizing material

Mean property value Mean and variability Comparison with specification Comparison of batches Identification of “hot spots” Presence / absence

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

84

Sampling purpose and material characterization

How much is in the plant? How much is in each fraction of the plant? How much is in the entire harvest? Is there a difference between greenhouses?

Is there a difference between the levels in the greenhouse? Is there a difference between spring and winter harvest?

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

85

Growing, harvesting, drying – where do we start?

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

86

Sample mass

Depends on:

Quantity of bulk material Heterogeneity Composition Variability Sample mass for analysis

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

87

How many increments?

More smaller increments is better than fewer larger Minimum number of increments to collect

= 10, 3 =

– min number of increments to sample - total number of increments in bulk - mass of bulk material - mass of increment

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

88

Sampling designs

Types

Designs

Probability based Objective ` Judgmental Combined

Simple random Systematic Stratified Composite

Controlling probability is the best super-power. And I think there’s a big chance that you’ll agree

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

89

Simple Random Sampling

Simple random sampling Use random number generator to define selected units

NEPTUNE AND COMPANY, INC improving the quality of decision making

90

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

Random selection of units:

20, 14, 28, 17, 30, 26, 8, 16, 3, 9,

Sampling Strategies

Systematic sampling

Set sampling interval n syst Randomly select starting point n 1

91

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

Interval:

every 5 th unit

Starting point:

,

Sampling Strategies

Stratified random sampling Use random number generator to define selected units

NEPTUNE AND COMPANY, INC improving the quality of decision making

92

1 2 3 4 5 6 7

9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

8

Random selection of units within a segment:

10, 1, 7

19, 12, 14

30, 24, 27

Sampling Strategies

Composite sampling Use random number generator to define selected units

NEPTUNE AND COMPANY, INC improving the quality of decision making

93

1 2 3 4 5 6 7

9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

8

Random selection of units within a segment:

10, 1, 7

19, 12, 14

30, 24, 27

Sampling from drying racks

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

94

Sampling from large containers and bags

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

95

Sampling from trays

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

96

Sampling products

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

97

7 Sampling Errors per Gy Theory

1. Fundamental error 2. Grouping and segregation 3. Large-scale heterogeneity 4. Periodic heterogeneity 5. Increment delineation 6. Increment extraction 7. Bad preparation

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

98

Quality Control

Duplicate samples Blind duplicate samples Split samples Equipment blank and equipment rinsate Archive samples

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

99

Q & A

Polona Carson, MBA, PhD pcarson@neptuneinc.org

NEPTUNE AND COMPANY, INC improving the quality of environmental decision making

100

Julie Kowalski, JA Kowalski Science Support LLC

Julie Kowalski is a technical consultant serving primarily the cannabis, hemp and food testing markets. She partners with clients offering technical expertise and advise taking advantage of her over 20 years of laboratory experience. She specializes in developing fit-for-purpose technical programs and customized training for laboratory staff. Julie has a passion for analytical chemistry, training and the cannabis testing industry, starting work on cannabis testing in 2011. Julie is a sought-after speaker and trainer focusing on educational presentations on various aspects of analytical chemistry.

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

Review of AOAC SMPR ® 2018.011 and A Possible Path Forward through Community Method Development ‘Think Tank’

Julie Kowalski, PhD Co-chair Cannabis Chemistry Working Group

August 29, 2023 AOAC INTERNATIONAL Annual Meeting

AOAC SMPR ® 2018.011 Standard Method Performance Requirements (SMPRs ® ) for Identification and Quantitation of Selected Pesticide Residues in Dried Cannabis Materials

Identification and Quantitation of Selected Pesticide Residues in Dried Cannabis Materials

Identification and Quantitation of Selected Pesticide Residues in Dried Cannabis Materials

Identification and Quantitation of Selected Pesticide Residues in Dried Cannabis Materials

Identification and Quantitation of Selected Pesticide Residues in Dried Cannabis Materials

Identification and Quantitation of Selected Pesticide Residues in Dried Cannabis Materials

Identification and Quantitation of Selected Pesticide Residues in Dried Cannabis Materials

Identification and Quantitation of Selected Pesticide Residues in Dried Cannabis Materials

104 compounds

104 compounds

0.005-1 ppm (mg/kg)

104 compounds

104 compounds

Challenges:

• Comprehensive compound list • Different regulatory limits • Changing/new regulations • Reporting rules

Challenges:

• Comprehensive compound list • Different regulatory limits • Changing/new regulations • Reporting rules

6 years later…

Pesticide Method “Think Tank” • Community-based, step-by-step method development • Share expertise, experience & best practices to draft a method • Evaluation of proposed method by volunteer labs

• provide feedback to the group • technical guidance documents?? • Validation of the optimized method • Submit method for ERP evaluation

We need your feedback & participation

Julie Kowalski julie@kowalskiscience.com

Joe Konschnik, Plus One Consulting LLC Representing Restek Corporation

As Principal Consultant for Plus One Consulting LLC Joe applies his 40 years of analytical chemistry knowledge and experience to support his clients in the analytical testing industry. Prior to retiring from RESTEK in January, 2023 Joe was RESTEK’s Business Development Manager for the global Food & Agriculture market where he identified market needs, new technologies, new product opportunities and coordinated technical content development for new workflow solutions. Prior to RESTEK Joe spent 19 years working for some of the nation’s largest environmental testing laboratory companies as a bench chemist and in technical and laboratory operations senior management roles. Joe has been an invited speaker on sampling and analytical chemistry topics globally. He serves as a volunteer in multiple AOAC working groups and communities and as a member of the TDRM Executive Committee. Joe has also previously served in volunteer leadership roles for the Independent Laboratories Institute (ILI), the American Council of Independent Labs (ACIL)’s Food Sciences Section and The North American Chemical Residue Workshop (NACRW). Direct correspondence at joe@plusone.llc, or joe.konschnik@restek.com.

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

Pesticide SMPR LOQs Proposal for Cannabis Joe Konschnik, Plus One Consulting LLC Representing Restek Corporation

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

Pesticide SMPR LOQs Proposal https://www.acil.org/

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

Pesticide SMPR LOQs Proposal www.acil.org/page/Cannabis_Working_Group

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

Pesticide SMPR LOQs Proposal 49 Page document Written by

• Analytical Chemists • Multiple Independent Cannabis Testing Labs, Consultants & Suppliers • Reviewed by Assessors, AOAC’s ALACC, State Government Regulators, Consultants and ACIL CWG members

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

Pesticide SMPR LOQs Proposal First 17 pages cover these topics: • Introduction • Scope • Definitions/Terminology • Laboratory QMS o Org & Scope of Work o Personnel Requirements o Facility and Security

o Service Requests, IFBs, Contracts o Method Verification/Validation o Sample Management/Receipt o Technical Records o Uncertainty o QC and Data Analysis o Reporting Results o Etc.

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA

Pesticide SMPR LOQs Proposal Last 32 Pages: Appendices A - H • Cannabinoid Analysis • Microbial Analysis

o Moisture Content Analysis o Residual Solvents Analysis o Terpene Analysis o Water Activity Analysis

• Heavy Metals Analysis • Pesticides & Mycotoxin Analysis

AOAC INTERNATIONAL 137 th Annual Meeting & Exposition August 25 – 30, 2023 • New Orleans, LA