Ric Cuchetto · 2018-12-28 · Measure 91 2016 - Oregon 420 dispensaries 50,000 growers 36,000...

Ric Cuchetto

Objectives

Appreciate some of the obstacles states face when developing pesticide protocols for cannabis

Political

Scientific

Market-based

Basic understanding of technologies available

Physical limits

Sensitivity

Human limits

Process Feedback and Control

2/15/2018 © 2017 The Cannabis Chemist ™ 2

OMMP July 1, 2016

66,880 patients

29,833 caregivers

Oregon Medical Marijuana Program

November 1998

Not for Profit patient providers:

Caregivers

Growers


Implementation of recreational sales in OR and WA have spawned regulations that overlap or eclipse existing medical policy.

OMMP OLCC, OHA

ORELAP

Classes of pesticides Carbamate

EPA 531 HPLC derivitized

Organochlorine EPA 8081

GC-ECD

Pyrethroid EPA 8270

GC-MS

Organophosphate EPA 8140

GC-FPD

EPA Method language GCMS for semi-volatile compounds 1980’s technology


GC-FID (for compliance?)

2/15/2018

Inaccuracies multiply at lower concentrations and with RT shift

“hydrocarbon burner”;non-specific detector

Thin Layer Chromatography

(TLC)Immunoassays

(ELIZA)Total Organic Halides

(TOX)

© 2017 The Cannabis Chemist ™ 5

This slide from Jack Cochran’s excellent blog at Restek

Lower cost technologies

GC: Co-extractives of cannabis


Thanks to Rodger Voelker, PhD

MS vs MS/MSsingle quad (GC or LC) cannot detect low levels in complex matrices

triple quad (GC or LC) can


Matrix effect becomes more pronounced as foodstuff gets more complex (less water, more oily)

Cannabis and hops behave more like spinach or citrus

Accurate ID and quantitation via MS/MS

Tandem Quad aka Triple Quad –The Typical Approach for QqQ

© 2017 The Cannabis Chemist ™ 8

Chromatogram of 195 Pesticides in Organic Honey

2/15/2018

Technology shift


2013 – GC MSD $36k (refurbished) 2016 – HPLC MSTQ $300k+

new language/disciplineGC LC

new technologyMSD TQ

New cost of entry

Pesticides – new targetsAbamectin

(Avermectins B1a and B1b Insecticide/acaricide LC-FLD1; LC-MS/MS2

Bifenazate Acaricide LC1; LC-MS/MS2

Bifenthrin

(synthetic pyrethroid) Insecticide GC-ECD1; GC-MS/MS2

Cyfluthrin

(synthetic pyrethroid) Insecticide LC2 (WHO 2004); GC-MS/MS2

Daminozide(Alar) Plant growth regulator (PGR); fungicide UV Spectroscopy1; LC-MS/MS2

Etoxazole Acaricide GC-MS(/MS)1

Fenoxycarb Insecticide LC/UV1; LC-MS/MS2

Imazalil Fungicide GC-ECD1; LC-MS/MS2

Imidacloprid Insecticide LC-MS/MS2

Myclobutanil Fungicide GC-ECD; GC-NPD1; GC-MS/MS)2; LC-MS/MS2

Paclobutrazol Plant growth regulator (PGR); fungicide LC-MS/MS2

Pyrethrins Insecticide GC-ECD1

Spinosad Insecticide LC-MS/MS; immunoassay1

Spiromesifen Insecticide GC-MS1; LC-MS/MS2

Spirotetramat Insecticide LC/LC-MS/MS2

Trifloxystrobin Fungicide GC-NPD1; GC-MS/MS2; LC-MS/MS2

Carbofuran

Diazinon

Malathion

Pesticide Use Residue Analytical Methods (RAM) Environmental

Protection Agency (EPA)1 or Literature2

10© 2017 The Cannabis Chemist ™2/15/2018

Compliance testing under OMMP

Potency THC, including THC acid CBD, including CBD acid CBN

Pesticides 20+ components

Mold/Mildew

% Moisture

HPLC MS/MSLC – QQQ


LC-MS pesticidesAbamectin(Avermectins B1a and B1bBifenazateBifenthrin(synthetic pyrethroid)Cyfluthrin

(synthetic pyrethroid)

Daminozide (Alar)

Etoxazole

Fenoxycarb

ImazalilImidaclopridMyclobutanilPaclobutrazol PyrethrinsSpinosad Spiromesifen Spirotetramat Trifloxystrobin

CarbofuranDiazinonMalathion


Measure 91 2016 - Oregon

420 dispensaries

50,000 growers

36,000 sites

34 labs

*Source: Oregon.gov

In order to implement Measure 91, the Oregon Liquor Control Commission

is required to establish a Rules Advisory Committee (RAC). The RAC is

charged with assisting in development of the fiscal impact statement and

helping the Commission develop the set of rules necessary to implement the

provisions of Measure 91.

Due to the complexity of the issues surrounding Measure 91, the OLCC will

establish technical committees of subject matter experts who will assist the

RAC and the OLCC in fully understanding the impact that the draft rules

will have upon specific areas of the regulated marijuana market.

Two Technical Committees; Licensing, Compliance & Enforcement, and

Labs will be ongoing to educate and inform the RAC and OLCC staff on

issues and topics in the recreational market.


Unique sources of contamination

Products for ornamental use ONLY Roses have similar pests and bloom nutrients

Nutrient supplements Non labeled additives

CBD from Hemp Pesticides not allowed in US

Cross-contamination Extraction (CO2, butane)

Soils; persistent (Myclobutanil, etc.)

Applicators “Mfr opinion”


Diversion – Cole Memo

Vice; like tobacco or alcohol

Price point for analytical

© 2017 The Cannabis Chemist ™ 152/15/2018

Venn diagrams to a practical path for safe cannabis

QuEChERs method has limitations

Electrospray Ionization (ESI) has limitations

HPLC required for many analytes

GC preferred for a few analytes

Certified cannabis labs; n = 0 Aug 2016


“Action Levels”

Levels at which the labs are to be certified

FIFRA is still law:

10ppb limit


No chemicals “approved” for use on cannabis

Tandem Quad is often the best solution…

Provides quantitative screening for known compounds for which there is a standard available

…but not always

Precise quantitative screening can be limited by:• Limited standard availability or elevated costs of uncommon compounds

• Infrequent occurrence of some compounds – why calibrate?

• Multiple matrix calibrations require time and resource

• ESI Bias for HPLC, Broad coverage via GC


Electrospray bias


100%

+/- 3x

no cleanup

0

5

10

15

20

25

30

AO

AC

Qu

EC

hE

RS

/no

cle

an

up

Un

bu

ffe

red

Qu

EC

hE

RS

/no

cle

an

up

Ace

ton

itri

le (

FA

) /n

o c

lea

nu

p

No

n-v

ola

tile

ma

teri

al

(mg

/mL

)Gravimetric Evaluation

Pass throughcleanup

dispersiveSPE

cleanup

Different sample preparations

Thanks to Julie Kowalski

5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5

Acetonitrile (0.1 % formic acid) no cleanup

AOAC QuEChERSPSA 50, C18 50, GCB 7.5 dSPE

AOAC QuEChERSPSA 100, C18 100, Chlorofiltr 50 dSPE

Acetonitrile (0.1 % formic acid) EMR Lipid blend

Acetonitrile (0.1 % formic acid)/ UCT pass through 2

Volatile Analysis (GC-MS)1g/10mL

1:20 dilution

Retention

time (min)

Pesticide Recovery Check

0

10

20

30

40

50

60

70

80

90

EMR Lipid blend

UCT pass through 2

69% and below 70% to 120%

% compounds in recovery ranges

30

12

70

88

% c

om

po

un

ds

QuECHERS?


dSPE LC protocol


dSPE GC protocols


QuECHERS?


Technology shift


2016 – LC TQ $300k (refurbished) 2018 – LC TQ $600k+GC TQ $250k

new language/disciplineQuEChERs targeted cleanup

high dilution

improved technologyTQ TQ

(LC-ESI GC-EI)(ESI APCI)

New cost of entry

Importance of Food SafetyMarket Landscape and Dynamics

Brand ProtectionGlobalizationPopulation growth

Pesticides analysis represents $400M of the total Food Safety market with a project growth of 6%. Cannabis growth is well into double digits.

To meet expected demand in 2030, Food production needs to increase by 70%

Nationalization of cannabis supply chain means producers must accommodate many differing state regulations

ISO 17025 supporting cGMP

Negative Press can have lasting effect on Brand success in nascent markets

Prop 65 levies

Interstate cosmetics


Compliance


Process Control + Feedback!

Extraction Process Butane

CO2

Ethanol

Fluorocarbon

Quality Control Dosing

Cannabinoids

Flavor/Fragrance


Identification/QuantitationCombined Workflow by TOF

TOF is a technique which can combine benefits of quantitation with qualitative screening.

A full spectrum, accurate mass approach allows:

• Reliable identification of compounds, even without a standard

• Possibility to look for new compounds

• Retrospective data analysis


Enhanced Selectivity using Accurate Mass

EIC: 333 (± 0.5 amu) S/N (peak to peak): 2.5

EIC: 362 (± 0.5 amu) S/N (peak to peak): 8.9

EIC: 333.9826 (± 20 ppm) S/N (peak to peak): 21.8

EIC: 362.0139 (± 20 ppm) S/N (peak to peak): 23.5

TIC

Narrow mass extraction window improves s/n ratio and selectivity

Coumaphos 5 ng/mL spiked in blended food matrices extract


Pesticide library

▪ Exact mass spectra for more than 850 pesticides and environmental contaminants

▪ Retention time locked screening methods

▪ Retention times help verify hits as another measure of confirmation in addition to exact mass spectra

PCDL = Personal Compound Database and Library© 2017 The Cannabis Chemist ™ 342/15/2018

Technology shift?


2016 – LC TQ $300k (refurbished) 2018 – GC-TOF $600k+

new language/disciplineQQQ TOF

new technologyLC LC + GC

New cost of entry

New capabilitiesDiscovery/Isolations

MetabolomicsHuman -0mics

Conclusions Data tsunami

Broad based extraction and analysis

200 analytes @ 100ppb -> 700 analytes @ 10ppb

Targeted extractions and analyses

Cleanups

Platforms

Correct tools

Electronic filtering of data (workflow)

200 compounds x 50 samples/day x 3 ions = 30,000 data points

Retention time and ion ratios

Lower Limits of Quantitation (LOQ) approaching 10 ppb


Future Scanning analyses for Tentatively Identified Compounds

Spectroscopy or Chromatography? Accurate Mass

LCxLC, GCxGC

Labelled internal standards for critical/problem analytes Class based or time based?

Bifenthrin

Imidacloprid

Accreditation; not just licensing Proficiency Test samples (in cannabis)

Primacy Labs/Methods AOAC


Acknowledgements

Rodger Voelker

Rick Jordan/Steve Thun

Julie Kowalski

courageous public servants LC QQQ posse

2/15/2018 38© 2017 The Cannabis Chemist ™

Questions?

39© 2017 The Cannabis Chemist ™

(503) 403-8579

[email protected]/Facebook – The Science of Flower

2/15/2018

Ric Cuchetto · 2018-12-28 · Measure 91 2016 - Oregon 420 dispensaries 50,000 growers 36,000...

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Transcript of Ric Cuchetto · 2018-12-28 · Measure 91 2016 - Oregon 420 dispensaries 50,000 growers 36,000...