Importance of Safety Compliance Facilities

• They will be the first to get licensed

• The most important difference in a regulated industry

• The state called them “safety compliance” facilities for a reason. They did not call them testing facilities

• No Financial Interest in another licensee type

• Is authorized to test for caregivers

Caregivers for Life Colorado

• January 2016 Voluntary Recall of 2,300 Units of Oil

• Blamed their suppliers of trim

• Lost Reputation

• Plummet in Revenue and Profit

Kindman Cultivation Facilities (MCI, Inc.)

• Dozens of Strains recalled and held due to identification of unapproved pesticides

• Plants grown from March 2014 through March of 2016

• Passed original testing but was then investigated by the Colorado Department of Agriculture based upon a tip of the use of a banned pesticide

• Traces as low as 5 parts per billion (ppb) were identified

Colorado Pesticide Recalls• 16 Recalls in 2015

• 23 Recalls in 2016

• 9, so far, in 2017 (through 8/21/17)

• 49 % of cannabis samples tested identified residue of unapproved pesticides last year, 13% so far this year

• http://www.thecannabist.co/2015/12/04/pesticide-pot-recall-list-marijuana/44711/

Oregon Pesticide Recalls• Hundreds of samples identified pesticides last year

and failed.

• Three pesticides accounted for the more than 900 cannabis failures since October of 2016.

• spinosad

• piperonyl butoxide

• myclobutanil - Eagle20

Cannabis Testing

Identification and Quantification of components in the following categories

CannabinoidsPsychoactive and non-psychoactive compounds produced in the Cannabis plant

TerpenesAromatic constituents of Cannabis resin and essential oils

Residual SolventsResidual solvents for extracts made with hydrocarbons or organic solvents

PesticidesPesticide residues

Trace MetalsToxic heavy metals such as arsenic, cadmium, lead, and mercury

Microbiological ImpuritiesDetection and quantitation of relevant bacterial and fungal species

Water ActivityMeasure of water available to support microbial growth

Cannabis Testing – Cannabinoids

Cannabinoids - Potency testingPsychoactive and non-psychoactive compounds produced in the Cannabis plant. Cannabinoids have similar chemical structure but very different effects. Concentration is measured in percent (e.g., THC 2-20% in plant material) or in mg/g (10% = 100 mg/g).

THC - Delta-9 Tetrahydrocannabinol

THCA - Tetrahydrocannabinolic Acid

CBD - Cannabidiol

CBDA - Cannabidiolic Acid

CBN - Cannabinol

CBG - Cannabigerol

The acid forms will transforminto the neutral form when heated (e.g. THCA > THC).

Analysis performed withHigh Pressure LiquidChromatography (HPLC) orGas Chromatography (GC).GC will heat the sample.

Cannabis Testing – Solvents

Residual SolventsLeftover chemicals from the extraction process to prepare oils and waxes. Those are harmful to humans and need to be removed by evaporation.

AcetoneBenzenen-ButaneIsobutaneEthanolEthylacetateHeptaneIsopropanolMethanolPentanePropaneTrichloromethaneTrichloroethyleneToluene

Analysis performed withHead Space GasChromatography (GC).

Concentration levels need to be measuredat the level of 1 to 1000 ppm (parts per million),depending on the toxicity of the solvent.

1 ppm = 1 mg / kg(approx. a grain of sugar in a pound of salt)

Cannabis Testing – Terpenes

Terpene ProfilingTerpenes are aromatic compounds in Cannabis resin and essential oils. Terpenes give cannabis its distinctive flavor and aroma.

Alpha-Pinenepine

Beta-Caryophyllenepepper, clove

Borneolcamphor

Caryophyllene oxideeucalyptus

Cineoltea tree

Citronellolrose

Phytolgreen tea

Linaloollavender

Humulenehops

Limonenecitrus

Myrcenelemongrass

Terpinolenelilac

Analysis performed by separation with Gas Chromatography and subsequent identificationwith Mass Spectrometry(GCMS).

Cannabis Testing – Pesticides

Pesticide ResiduesThe screening for pesticide residue requires highly sensitive measurement equipment with very low detection limits. Special extraction methods need to be used to extract pesticides for analysis from the plant material.Abamectin Acephate Acequinocyl Acetamiprid Aldicarb Azoxystrobin Bifenazate Bifenthrin Boscalid Captan Carbaryl Carbofuran Chlorantraniliprole Chlordane Chlorfenapyr Chlorpyrifos Clofentezine Coumaphos Cyfluthrin Cypermethrin Daminozide DDVP (Dichlorvos) Diazinon Dimethoate Dimethomorph

Ethoprop(hos) Etofenprox Etoxazole Fenhexamid Fenoxycarb Fenpyroximate Fipronil Flonicamid Fludioxonil Hexythiazox Imazalil Imidacloprid Kresoxim-methyl Malathion Metalaxyl Methiocarb Methomyl Methyl parathion Mevinphos Myclobutanil Naled Oxamyl Paclobutrazol Pentachloronitrobenzene

Permethrin Phosmet Piperonyl butoxide Prallethrin Propiconazole Propoxur Pyrethrins Pyridaben Spinetoram Spinosad Spiromesifen Spirotetramat Spiroxamine Tebuconazole Thiacloprid Thiamethoxam Trifloxystrobin

Analysis performed withLiquid ChromatographyMass Spectrometry (LCMS) orGas ChromatographyMass Spectrometry (GCMS).

Concentration levels need to be measuredat the level of 10 to 1000 ppb (parts per billion).depending on the toxicity of the solvent.

1 ppb = 1 ug / kg(approx. a grain of sugar in a thousand pounds of salt)

Microbiological• Mites/Bugs

• Mold

• Bacterial -

• Escherichia Coli

• Salmonella

• Aspergillus

• Mycotoxins

• Foreign Material

Heavy Metals

• Expensive Instrument ($160,000.00)

• Some States Require it, many do not

• No Present Testing facilities have a ICP-MS

Trace Metal Testing – ICPMS

Inductively Coupled Plasma Mass Spectrometry (ICPMS) ICPMS is a highly sensitive method that can be used to analyze trace amount of metals. ICPMS can

measure concentrations as low as 1 ppt (1 part per trillion). It uses an argon plasma torch that heats

the sample to up to 10,000 degree Celsius (18,000 F).

Metals in the sample are converted

into charged ions and analyzed in a

high resolution mass spectrometer.

Portion of an ICP mass spectrum showing the

presence of mercury and lead in a sample.

(Shimadzu Cannabis Testing Solution Brochure)

The Federal Pesticide Problem

• EPA regulates and licenses all pesticides

• Products Must be Used According to Label

• Broad Label and Below Tolerance Products

Required Oregon Analytes

• Pesticides

• Water Activity and Moisture Content

• THC and CBD

• Microbiological only upon request

• Solvents (processor only)

Colorado Analytes (Retail)• Potency

• Microbiological (Bacteria and Fungus)

• Residual Solvents

• Metals

• Pesticides

• Anything injurious to human health

Colorado Potency Testing

• A single percentage concentration for each cannabinoid must represent an average

• Every individually packaged THC processed item shall have no more that 100mg THC per volume

• Potency Variance of +/- 15%

Homogeneity Testing

• Ensures that any item being tested contains equal amounts of the analyte in every location of the individual item

• Manufactured or processed edible products - spread throughout the product evenly and not concentrated in one area

Colorado Homogenous Requirements

• A 10% portion of the THC infused or manufactured product cannot contain more than 20% of the total THC within the entire product

Colorado Microbial Testing

• Shiga-toxin producing Echerichia Coli

• Salmonella

• Total Yeast & Mold

• less than 1 Colony Forming Unit (CFU)

• less than 1 CFU

• less than 10(4) CFU

Colorado Metals Testing

• Lead

• Arsenic

• Cadmium

• Mercury

1 PPM

.4 PPM

.4 PPM

.2 PPM

Colorado Residual Solvent Testing

• Butanes

• Heptanes

• Benzene

• Toluene

• Hexane

• Total Xylenes

5,000 Parts Per Million

5,000 PPM

2 PPM

890 PPM

290 PPM

2,170 PPM

Required California Analytes• Cannabinoid Potency

• Residual Solvents

• Residual Pesticides

• Microbiological

• Mycotoxins

• Water Activity/Moisture Content

• Foreign Material

• Heavy Metals

• Terpenes

California Residual Solvents Testing

1,2-Dichloroethane

Acetone

Acetonitrile

Benzene

Butane

Chloroform

Ethanol

Ethyl acetate

Ethyl ether

Ethylene oxide

Heptane

Hexane

Isopropyl alcohol

Methanol

Methylene chloride

Naphtha

Pentane

Petroleum ether

Propane

Trichloroethylene

Toluene

Total xylenes

California Homogenous Requirements

• No less than a 15% standard deviation between two different samples of the same item

3/1/16%

4%

A2LA Cannabis Program January 2014

•  Reluctant entrance   Initially we considered just medicinal  Concerns over federal banking issues &legal liability   Unsure of market size: ~20 states with various programs,

few labs

•  Research showed strong trending nationally

Laboratory Accreditation

Accreditation is a formal recognition by an authoritative third-party of a laboratory’s competence to perform specific tests.

ISO 17025 is an international standard defining the general requirements for the competence of testing and calibration laboratories.

The requirements are based on best-practices and the general scientific consensus. They are written in a general way so that they apply no matter what quantity is being measured.

Accreditation doesn’t guarantee reliable results but ensures that the lab has the competence and the processes in place that are needed to achieve reliable results.

INTERNATIONAL STANDARD

ISO/IEC17025Second edition

2005-05-15

General requirements for the competence of testing and calibration laboratories

Exigences générales concernant la compétence des laboratoires d'étalonnages et d'essais

It is better to have no tests than wrong or misleading results!

Laboratory Accreditation – ISO 17025

Elements of Technical Requirements

Education, training, and skills of personnel

Environmental conditions in the lab

Procedures for testing, including sampling, handling, transport, and preparation of items

Validation of methods

Estimation of Uncertainty of Measurement

Measurement Traceability

Control of Equipment

Quality Control Procedures

Reporting of Results

Management Requirements

Management requirements are similar to other Quality Management Systems (ISO 9001):Control of documents and recordsCustomer servicesCorrective Action processInternal auditsManagement reviews

INTERNATIONAL STANDARD

ISO/IEC17025Second edition

2005-05-15

General requirements for the competence of testing and calibration laboratories

Exigences générales concernant la compétence des laboratoires d'étalonnages et d'essais

No ISO 17025 Accreditation Required

• Oregon (Requires accreditation through the Oregon Environmental Accreditation Program (ORELAP) which utilizes the TNI Laboratory Standards developed by the National Environmental Laboratory Accreditation Conference

• CA developed their own standards for testing laboratories

3/1/16%

6%

Variety of Quality Management Standards

•  Washington: WSLCB GLP •  Oregon: ORELAP •  Colorado: No mention/ Dept. of Revenue, MJ Enforcement Division •  Alaska: No mention •  New York: DEA certified and NYS ELAP / Connecticut: CS registered •  Illinois: accredited by a private laboratory accrediting organization •  California: ISO 17025 as prerequisite to state recognition •  Maine: (in legislature) (ISO 17025/ A2LA) •  Maryland: ISO 17025 as prerequisite to state recognition •  Minnesota: encourages ISO 17025 / A2LA •  New Hampshire: ISO 17025 or CLIA as prerequisite to state recognition

Without industry consensus on QMS: Is this a Reliable ISO 17025 certification?

Method Validation

Method validation is the process of establishing the performance characteristics and limitations of a method.

It is an important element of quality control and to achieve measurement reliability.

Elements of Method Validation:Limit of DetectionLimit of QuantizationMeasurement UncertaintyInterference StudiesCarryover (cross-sample contamination)Calibration Model (linear range)

Validation and Peer Review of U.S. Environmental Protection Agency

Chemical Methods of Analysis

Prepared for:

The EPA Forum on Environmental Measurements (FEM)

Prepared by:

The FEM Method Validation Team

FEM Document Number 2005-01

October 14, 2005

REVISION: February 3, 2016

The Bottom Line

Scientifically Reliable Results

ValidationMethod Validation

(LOD, LOQ,Uncertainty,...)

TraceabilityTraceability to (inter)national

Standard

TransparencyTransparency of the Measurement

Process

Sampling• If a Representative Sample is not obtained, the

reliability and accuracy of the testing does not matter

• In California, Massachusetts, and Oregon the laboratory selects and transfers the sample

• Maryland requires an independent sampling company

• In Colorado the grower collects the samples (check)

Sampling

Challenge: Inhomogenous SamplesCannabinoids are highly lipophilic and disolve in oils or fat rather than in water.

Cannabinoid concentrations might significantly vary within samples that consist of watery and oily phases.

A Laboratory sample plan needs to address this situation and provide clarity to the customer.

Oil and Water don’t mix!

Gummi Bears are made froma water-based mixture of sugar, starch, flavor, color, and gelatin.

What is there is a failing test

• Product Recall

• Product Destruction

• Product Return for Remediation

• Litigation