Overcoming the Challenges Associated with Pesticide Analysis of Cannabis and
Understanding the Matrix Effects and the Impact of Various Clean-Up Strategies
Toby Astill, Ph.D. - PerkinElmer Emerald Conference March 1st 2019
Agenda:
• LC/MS-MS Mass Spectrometry Options
• Pesticide Analysis in Cannabis Distillate in California
• Pesticide Analysis in Cannabis Flower in Canada
• Learning & Collaborative Opportunities
• Summary
Cannabis is a Exciting/Challenging Matrix
LC/MS-MS Mass Spectrometry Options
• ESI – Electrospray ionization
• APCI - Atmospheric pressure chemical ionization • (H2O)2H+ - transfers the charge
[1] Deibel Cannabis Labs
California
California Pesticide & Mycotoxin Analysis
10x vs. 20x
Matrix Matched Suppression
California Method Matrixed Matched
California TIC @ 0.25ppb (250ppt)
Testing Flower
• ACN Extraction
• 10 Internal Stds
• 24min Run
Atrazine-d5Carbaryl-d7Daminozide-d4Diazinon-d10Dichlorvos-d6
Dimethoate-d6Imidacloprid-d4Myclobutanil-d9Pyridaben-d13Thiamethoxam-d3
Testing Concentrate & Distillate – why?
• Concentrates may surpass flower as the #1 product in the US
Sample Prep
Distillate
Flower
ACN
ACN
The data doesn’t lie…. 100 ppb PCNB Spike
Why Suppression?
CBCTHCV
CBG/CBDCBN
Δ8 THC
CBDV
Δ9 THC
Internal Standards An internal standard is a refence standard that is added in a constant amount to samples, the blank and calibration standards in a chemical analysis.
Internal Standard Data
0
20
40
60
80
100
120
0 100 200 300 400 500 1000
% re
cove
ry
Sample wt (mg)
Myclobutanil
myclobutanil -d9 as IS no IS
0
20
40
60
80
100
120
0 100 200 300 400 500 1000
% R
ecov
ery
Sample wt (mg)
Diazinon
Diazinon-d10 as IS no IS
0
20
40
60
80
100
120
140
160
0 100 200 300 400 500 1000
% R
ecov
ery
Sample wt (mg)
Pyridaben
pyridaben-d13 as IS no IS
Internal Standard DataMyclobutanil with IS
Sample wt (mg)
Spiked Concentration (ppb)
Observed Concentration (ppb)
Percent Recovery
0 100 102 102%100 100 103 103%200 100 106 106%300 100 103 103%400 100 104 104%500 100 104 104%
1000 100 111 111%
Myclobutanil no IS
Sample wt (mg)
Spiked Concentration (ppb)
Observed Concentration (ppb)
Percent Recovery
0 100 112 112%100 100 105 105%200 100 98 98%300 100 80 80%400 100 75 75%500 100 69 69%
1000 100 53 53%
Diazinon with IS
Sample wt (mg)
Spiked Concentration (ppb)
Observed Concentration (ppb)
Percent Recovery
0 100 99 99%100 100 97 97%200 100 90 90%300 100 93 93%400 100 89 89%500 100 88 88%
1000 100 86 86%
Diazinon no IS
Sample wt (mg)
Spiked Concentration (ppb)
Observed Concentration (ppb)
Percent Recovery
0 100 102 102%100 100 93 93%200 100 91 91%300 100 79 79%400 100 67 67%500 100 64 64%
1000 100 49 49%
Pyridaben with IS
Sample wt (mg)
Spiked Concentration (ppb)
Observed Concentration (ppb)
Percent Recovery
0 100 94 94%100 100 76 76%200 100 58 58%300 100 62 62%400 100 66 66%500 100 95 95%
1000 100 152 152%
Pyridaben no IS
Sample wt (mg)
Spiked Concentration (ppb)
Observed Concentration (ppb)
Percent Recovery
0 100 93 93%100 100 37 37%200 100 23 23%300 100 18 18%400 100 14 14%500 100 14 14%
1000 100 10 10%
Not all ‘I.S.’ created equal
Ionization Mode Analyte/Quantifier Transition IS for Quantifier Spiked Concentration
(ppb)Observed Concentration for
Blank Spike (ppb)Observed Concentration for Spiked Sample (ppb)
positive Hexythiazox (353.1/228) Diazinon-D10 100 125 16
positive Hexythiazox (353.1/228) Pyridaben-D13 100 97 61
positive Spirotetramat (374.2/216) Mycycobutanil-D9 100 96 235
positive Spirotetramat (374.2/216) none 100 96 124
positive Ethoprophos (243.1/131) none 100 110 36
positive Ethoprophos (243.1/131) Mycycobutanil-D9 100 94 74
Oh Canada!
Canada’s ‘96’
……10ppb
Source Optimization
Abamectin is Sensitive to Heat in Ion Source
0.00E+005.00E+041.00E+051.50E+052.00E+052.50E+053.00E+053.50E+054.00E+054.50E+055.00E+05
50 100 150 200 250
Abam
ectin
Sig
nal
Source Temperature/oC
100ppb Abamectin MRM Quantifier
S/N=67
The sensitivity was improved for abamectin by a factor of 10X(LOQ=25 ppb)
Stability Data for Dimethoate in Cannabis Extract
0.E+005.E+041.E+052.E+052.E+053.E+053.E+054.E+054.E+055.E+055.E+05
0 50 100 150 200 250 300 350 400 450 500
100
ppb
Dim
etho
ate
Peak
Are
a Ra
tio
to In
tern
al st
anda
rd in
Can
nabi
s Ex
trac
t
Run Number
RSD = 5.74 %
0.00
0.05
0.10
0.15
0.20
0.25
0.30
0 50 100 150 200 250 300 350 400 450 500
100
ppb
Dim
etho
ate
Peak
Are
a Ra
tio
to In
tern
al st
anda
rd in
Can
nabi
s Ex
trac
t
Run Number
RSD =1.9 %
No Internal Standard
Internal Standard
Finished Flower MethodS. No. Anayte Dried Marijuana Cannabis oil Qsight LOQ Ratio of Action Limits to Qsight LOQ
1 Abamectin 0.5 ppm 0.25 ppm 0.025 ppm 202 Acephate 0.02 ppm 0.05 ppm 0.010 ppm 23 Acetamiprid 0.1 ppm * 0.005 ppm 204 Acequinocyl * * 0.025 ppm NA5 Aldicarb 1 ppm 0.5 ppm 0.005 ppm 2006 Allethrin 0.2 ppm 0.1 ppm 0.050 ppm 47 Azadirachtin 1 ppm 0.5 ppm 0.05 ppm 208 Azoxystrobin 0.02 ppm 0.01 ppm 0.001 ppm 209 Benzovindiflupyr 0.02 ppm 0.01 ppm 0.003 ppm 6.67
10 Bifenzate 0.02 ppm 0.01 ppm 0.002 ppm 1011 Beifenthrin * 0.5 ppm 0.010 ppm NA12 Boscalid 0.02 ppm 0.01 ppm 0.005 ppm 413 Buprofenzin 0.02 ppm 0.01 ppm 0.002 ppm 1014 Carbaryl 0.05 ppm 0.025 ppm 0.004 ppm 12.515 Carbofuran 0.02 ppm 0.01 ppm 0.001 ppm 2016 Chlorantraniliprole * * 0.003 ppm NA17 Chlorphenapyr * 1.5 ppm 0.050 ppm NA18 Chlorpyrifos * 0.5 ppm 0.005 ppm NA19 Clofentezine 0.02 ppm 0.01 ppm 0.005 ppm 520 Clothiandin 0.05 ppm 0.025 ppm 0.003 ppm 16.6621 Coumaphos 0.02 ppm 0.01 ppm 0.002 ppm 1022 Cyantranilipole 0.02 ppm 0.01 ppm 0.002 ppm 1023 Cyfluthrin * 3.75 ppm 0.25 ppm NA24 Cypermethrin * 3.75 ppm 0.1 ppm NA25 Cyprodonil * 0.01 ppm 0.005 ppm NA26 Daminozide * * 0.015 ppm NA27 Deltamethrin * 12.5 ppm 0.010 ppm NA28 Diazinon * 0.5 ppm 0.003 ppm NA29 Dichlorvos 0.1 ppm 0.05 ppm 0.01 ppm 1030 Dimethoate 0.02 ppm 0.01 ppm 0.002 ppm 1031 Dinotefuran 0.1 ppm 0.05 ppm 0.005 ppm 2032 Dodemorph * * 0.005 ppm NA33 Endosulfan-Alpha * 2.5 ppm 0.05 ppm NA
Finished Flower MethodS. No. Anayte Dried Marijuana Cannabis oil Qsight LOQ Ratio of Action Limits to Qsight LOQ
34 Endosulfan-beta * 2.5 ppm 0.05 ppm NA35 Endosulfan-sulfate * 2.5 ppm 0.04 ppm NA36 Ethoprophos 0.02 ppm 0.01 ppm 0.002 ppm 1037 Etofenprox * * 0.004 ppm NA38 Etoxazole 0.02 ppm 0.01 ppm 0.01 ppm 2039 Etridiazole * 0.15 ppm 0.033 ppm NA40 Fenoxycarb 0.02 ppm 0.01 ppm 0.002 ppm 1041 Fenpyroximate 0.02 ppm 0.01 ppm 0.002 ppm 1042 Fensulfothion 0.02 ppm 0.01 ppm 0.002 ppm 1043 Fenthion * 0.01 ppm 0.003 ppm NA44 Fenvalerate * 3.75 ppm 0.5 ppm NA45 Fipronil 0.06 ppm 0.01 ppm 0.010 ppm 646 Flonicamid 0.05 ppm 0.025 ppm 0.005 ppm 1047 Fludioxinil 0.02 ppm 0.01 ppm 0.01 ppm 248 Fluopyram 0.02 ppm 0.01 ppm 0.004 ppm 549 Hexythiazox * * 0.002 ppm NA50 Imazalil 0.02 ppm 0.01 ppm 0.005 ppm 451 Imidacloprid 0.02 ppm 0.01 ppm 0.002 ppm 1052 Iprodione 1 ppm 1.5 ppm 0.050 ppm 2053 Kinoprene * 1.25 ppm 0.05 ppm NA54 Kresoxim-methyl * 0.15 ppm 0.010 ppm NA55 Malathion 0.02 ppm 0.01 ppm 0.005 ppm 456 Metalaxyl 0.02 ppm 0.01 ppm 0.002 ppm 1057 Methiocarb 0.02 ppm 0.01 ppm 0.002 ppm 1058 Methomyl 0.05 ppm 0.025 ppm 0.005 ppm 1059 Methoprene * 5 ppm 0.05 ppm NA60 Methyl Parathion * * 0.040 ppm NA61 Mevinphos 0.05 ppm 0.025 ppm 0.003 ppm 16.6662 MGK-264 * * 0.010 ppm *63 Mycyclobutanil 0.02 ppm 0.01 ppm 0.005 ppm 464 Naled * * 0.005 ppm NA65 Novaluron 0.05 ppm 0.025 ppm 0.003 ppm 16.6666 Oxamyl 3 ppm 1.5 ppm 0.005 ppm 600
Finished Flower MethodS. No. Anayte Dried Marijuana Cannabis oil Qsight LOQ Ratio of Action Limits to Qsight LOQ
67 Pacalobutrazol 0.02 ppm 0.01 ppm 0.005 ppm 468 Permethrin * 2.5 ppm 0.010 ppm NA69 Phenothrin 0.05 ppm 0.25 ppm 0.010 ppm 570 Phosmet * * 0.005 ppm NA71 Piperonyl Butoxide * 1.25 ppm 0.002 ppm NA72 Pirimicarb 0.02 ppm 0.01 ppm 0.001 ppm 2073 Prallethrin * * 0.005 ppm NA74 Propiconazole * * 0.010 ppm NA75 Propoxur 0.02 ppm 0.01 ppm 0.002 ppm 1076 Pyraclostrobin 0.02 ppm 0.01 ppm 0.004 ppm 577 Pyrethrins 0.05 ppm 0.025 ppm 0.025 ppm 278 Pyridaben 0.05 ppm 0.025 ppm 0.002 ppm 2579 Quintozene * 0.5 ppm 0.01 ppm NA80 Resmethrin 0.1 ppm 0.05 ppm 0.010 ppm 1081 Spinetoram * 0.01 ppm 0.01 ppm NA82 Spinosad * 0.01 ppm 0.01 ppm NA83 Spirodiclofen 0.02 ppm 0.01 ppm 0.01 ppm 284 Spiromesifen * 0.5 ppm 0.002 ppm NA85 Spitotetramat 0.02 ppm 0.01 ppm 0.002 ppm 1086 Spiroxamine * * 0.002 ppm NA87 Tebuconazole * * 0.010 ppm NA88 Tebufenozide 0.02 ppm 0.01 ppm 0.005 ppm 489 Teflubenzuron 0.05 ppm 0.025 ppm 0.005 ppm 1090 Tetrachlorvinphos 0.02 ppm 0.01 ppm 0.001 ppm 2091 Tetramethrin 0.1 ppm 0.05 ppm 0.010 ppm 1092 Thiacloprid 0.02 ppm 0.01 ppm 0.003 ppm 6.6693 Thiamethoxam 0.02 ppm 0.01 ppm 0.003 ppm 6.6694 Thiophanate-methyl 0.05 0.025 0.002 ppm 2595 Trifloxystrobin 0.02 ppm 0.01 ppm 0.001 ppm 20
Education & Engagement
Thanks to all those involved…
Summary• LC-MS/MS can be utilized for testing all pesticides in cannabis
• Cannabis Flower and Distillate must be studied independently
• Internal Standards are effective but selective
• ESI and APCI can quantitate both GC and LC amenable pesticides
• Robust and Stable APCI/ESI method with minimal prep and cleanup required
• One Instrument Solution for Pesticides Analysis in cannabis
• Recovery in the range of 70 -120 %
