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Barry K Logan PhD, DABFT
Director of Forensic Services
NMS Labs online Seminar
September 16, 2010
K2 and the Synthetic
Cannabinoids:
Pharmacology, Effects,
and Chemical Analysis
From the Premium Provider of Forensic Services
K2 Phenomenon
•Origins of synthetic
cannabinoid agonists.
•Contents of various.
commercial “K2” type
products.
•Mechanism of Action and
Pharmacology.
•Analytical challenges.
•Human
pharmacodynamics.
•A preview of the DUI
environment.
Marijuana
Most popular recreational drug after alcohol
and tobacco.
Main psychoactive component THC
#1 Drug in the DRE program
Some 25 million Americans have smoked
marijuana in the past year, and more than
14 million do so regularly.
Possession and use illegal under federal law,
but states have variable policies on
enforcement and prosecution.
6.8% of Friday and Saturday evening drivers
test positive for use.
Synthetic Cannabinoids
Synthetic drugs that mimic the
effects of cannabis.
Investigational use for appetite,
blood pressure, nausea, etc
First synthesized in the 1980’s
Sprayed onto dried plant leaves,
flowers and stems, and
smoked.
Legal or unregulated in most US
states.
K2 Phenomenon
Kansas,
Georgia,
Kentucky,
Alabama,
Tennessee,
Louisiana,
Missouri,
Hawaii,
Arkansas,
Illinois
Mississippi
JWH-018 JWH-073
Clemson Series - JWH
JWH-250
THC HU-210
CP 47,497
Marijuana to Spice
WIN 55,212
Cannabinoid Mechanism of Action
Cerebral Cortex
Higher Cognitive Function
Basal Ganglia
Cerebellum
Movement
Hippocampus
Learning, Memory, Stress
Hypothalamus
Appetite
Spinal Chord
Pain, Peripheral sensation
Medulla
Nausea, Vomiting
Herbals - Botanical Substrate
Glycyrrhiza glabra
(Cultivated Liquorice)
Astragalus membranaceus
(Milk Vetch)Verbascum thapsus
(Common Mullein)
(Uchiyama et al, 2010)
Synthetic Cannabinoids – Drug ID
• Agilent 5973 Mass Spectrometer
• Agilent 6890 Gas Chromatograph
• Electron Impact mode
• Full scan
• Total run time 10 minutes
• Sensitivity 20-50 ug/g
Synthetic Cannabinoids – Drug ID
JWH-018
JWH-073
JWH-200
HU-210/11
JWH-019
JWH-200
WIN 55,212
JWH-015
JWH-133
JWH-250
CP55,940
CP 47,497 (n=7)
CP 47,497 (n=8)
K2 Blends - Drug Content
(* Uchiyama et al, 2010)
JWH-018
(mg/g)
JWH-073
(mg/g)
CP47,497
(n=8) (mg/g)
JWH-250
(mg/g)
K2 Blonde 12 13 - -
K2 Standard 9 9 - -
K2 Citron 10 10 - -
K2* (Unknown) - - 6 -
K2 Summit 11 9 - -
K2 Blue 15 - - -
K2 Pink 11 - - -
K2 Latte 16 0.28 - 14
K2 Mint 19 0.30 - -
K2 Silver 8 - - 16
Spike Gold 20 11 - -
Spike Maxx 17 - - 19
Spike Diamond 17 0.07 - -
Spike Silver 9 16 - -
Space 10 - - -
Herbal blends* 2.0 – 35.9 - 1.1 – 16.9 -
Synthetic Cannabinoid Effects
T. Sobolevsky, et al., Detection of JWH-018 metabolites in smoking mixture post-administration urine,
Forensic Sci. Int. (2010), doi:10.1016/j.forsciint.2010.04.003
Synthetic Cannabinoids at NMS Labs
•Synthetic Cannabinoids in Blood - $195
•LCMSMS LOQ/LOQ - 0.1ng/mL
•JWH-018, JWH-073 (screened, confirmed, quantitated).
•JWH-018, JWH-073, JWH-019, JWH-250 (screened,
confirmed)
•Synthetic Cannabinoids in Urine - $60
•LCMSMS, LOD - 0.1ng/mL
•JWH-018 mono-hydroxy and di-hydroxy metabolites
(screened, confirmed)
•JWH-018 tri-hydroxy metabolites (monitored)
NMS Labs Special Projects
Matrix Analytes
Blood
Serum
Oral Fluid
(Refrigerate)
Parent Compounds –
JWH-018, JWH-073, JWH-200, HU-210, HU-211, JWH-019,
JWH-200 WIN 55,212, JWH-015, JWH-133, JWH-250,
CP55,940, CP 47,497 (n=7) CP 47,497 (n=8) …
Urine
(Freeze)Ring monohydroxylated metabolites – glucuronidated
Ring dihydroxylated metabolites – glucuronidated
Desmethyl ring hydroxylated metabolites - glucuronidated
Missouri K2 Administration Study
•IRB Human subjects and ethical approval obtained by
University of Central Missouri.
•Six subjects smoked K2 Summit, Citron, or Standard
•Each contained JWH-018, JWH-073, or CP47,497
•Subjects performed SFST’s, cognitive tests and DRE Exam.
•Blood, urine and oral fluid collected.
Missouri K2 Administration Study
•Onset of effects in about 2-3 minutes
•Dry mouth
•Light headedness
•Blurred vision
•Agitation, Motor restlessness
•Time dilation
•DRE Exam
•Increased pulse and blood pressure
•Lack of convergence
•No HGN, or VGN
•Pupils normal, muscle tone normal
•SFST’s
•3-4 inches of sway, leg body tremors
•Loss of balance
•Loss of motor coordination
Missouri K2 Administration Study
•Reported effects:
•Tachycardia (increased heart rate, 5-30mm/Hg)
•Dry Mouth
•Felt Impaired, subjective thought disruption
•Changes in perception
•Impaired sense of time
•Mild anxiety, paranoia
•Sedation
•Post-intoxication exhaustion
K2 use and the DRE Matrix
Drug Symptom Matrix
CNS Depressant Inhalants
Dissociative Drugs
CNS Stimulants Hallucinogens
Narcotic analgesics Cannabis K2
Horizontal nystagmus Yes Yes Yes No No No No No
Vertical Nystagmus Present * Present* Present No No No No No
Lack of Convergence Present Present Present No No No Present Present
Pupil Size Normal Normal Normal Dilated Dilated Constricted Dilated* Normal
Reaction to Light Slow Slow Normal Slow Normal Little to none Normal Normal
Pulse Rate Down Up Up Up Up Down Up Up
Blood Pressure Down Up/Down Up Up Up Down Up Up
Body Temperature Normal Up/Down/Normal up Up Up Down Normal Normal
* High doses
NMS Labs Blood JWH-018 and -073
L/L extraction
JWH-073 d-9 Istd
Quantitative LOD/LOQ 0.1ng/mL
Waters TQD Acquity
Gradient 4 minute run
Formic acid vs ACN
2 Transitions per analyte
NMS Labs Blood JWH-018 and -073
XIC of +MRM (34 pairs): 358.2/155.0 amu from Sample 3 (BM 2 hr Blood #... Max. 2.6e5 cps.
1 2 3 4 5 6 7 8 9Time, min
0.0
5.0e5
1.0e6
1.5e6
2.0e6
2.5e6
3.0e6
3.5e6
4.0e6
Inte
ns
ity
, c
ps
7.376.74
7.866.17
BM 2 hr Post dose blood
2 HR POST DOSE BLODD
IS
JWH-018
JWH-073
HO JWH-073
HO JWH-018
Workstation: NMS815
Printing Time: 4:53:36 PM
Printing Date: Wednesday, September 15, 2010
Electronic Signature: no
Analyst Version: 1.4.2
Page 1 of 1
Acq. Time: 10:26
Acq. Date: Friday, August 06, 2010
Acq. File: ED01_080610K2_AX.wiff
Sample Name: BM 2 hr Blood #2 not hydrolyzed
Sample Number: N/A
Batch Name: K2 080610.dab
Sample Comment:
Collected by: NMS815\labman
Sample ID: 3
Project: Production
Method Name: N/A
Results Name: N/A
Missouri K2 Administration Study
Subject BM
Smoked “K2 Citron”
10mg/g JWH-018/073
0.3g in water pipe
3 inhalations over 30 minutes
110
115
120
125
130
135
140
145
150
155
0
1
2
3
4
5
6
0:00 1:12 2:24 3:36
Systo
lic B
loo
d P
ressu
re (
mm
Hg
)
Co
nc
en
tra
tio
n (
ng
/mL
)
DT (hours)
JWH-018
JWH-073
Systolic BP
NMS Labs Urine JWH-018 and -073
L/L extraction
Qualitative
JWH-073 d-9 Istd
Method 1: hydrolyzed – Waters TQD
Mono and Di hydroxy metabolites
2 transitions per analyte
Method 2: unhydrolyzed – ABI 5000
Monohydroxy glucuronide – daughter – grandaughter
2 transitions per analyte and precursor
NMS Labs Urine SC metabolites
XIC of +MRM (35 pairs): 358.2/155.0 amu from Sample 2 (Hydrolyzed 19-2... Max. 4.2e6 cps.
2 4 6 8 10 12Time, min
0.0
5.0e5
1.0e6
1.5e6
2.0e6
2.5e6
3.0e6
3.5e6
4.0e6
4.2e6
Inte
ns
ity
, c
ps
6.80
7.33
6.003.47
JWH-018/073 Human Urinary Metabolites
IS
HO
HOCOOH
HO-N-dealkylation
2HO
3HO
Workstation: NMS815
Printing Time: 2:44:33 PM
Printing Date: Wednesday, September 15, 2010
Electronic Signature: no
Analyst Version: 1.4.2
Page 1 of 1
Acq. Time: 10:07
Acq. Date: Friday, September 03, 2010
Acq. File: ED01_090310K2_AX.wiff
Sample Name: Hydrolyzed 19-20-21 mix 1
Sample Number: N/A
Batch Name: K2 090310.dab
Sample Comment:
Collected by: NMS815\labman
Sample ID: 3
Project: Production
Method Name: N/A
Results Name: N/A
Missouri K2 Administration Study
Subject BM - Urine
Time JWH-018 Mono-OH Di-OH Tri-OH Glucuronides
Pre-dose X X X X X
1:15 +/- √ √ X √
2:07 X √ √ √ √
2:40 X √ √ √ √
Time JWH-073 Mono-OH Di-OH Tri-OH Glucuronides
Pre-dose X X X X X
1:15 X √ √ X √
2:07 X √ √ √ √
2:40 X √ √ √ √
York County PA, DRE Case
• Vehicle crashed into snowbank
• Subject appeared drunk or high, difficulty
speaking, incoherent, mumbling, slow slurred
speech, nonsense, giggling.
• Had smoked “Space” less than 2 hours before.
• “I’m not a DUI. The stuff I smoked is legal. You
can’t arrest me”
• Courtesy Cpl Brian Torkar, PSP
York County PA, DRE Case
• Pulse 96-88, BP 150/80
• Eyes watery, red, bloodshot, pupils dilated
• Eyelids droopy, HGN present, no VGN
• Lack of convergence
• SFST’s
• 4 inches of sway on Romberg
• Lost balance, used arms on WAT
• Swayed, used arms on OLS
• Poor finger to nose performance
• Courtesy Cpl Brian Torkar, PSP
York County PA, DRE Case
• Drug Identification
• “Space” contained JWH-018, traces of JWH-200
• Toxicology
• Blood collected approximately 4 hours after use.
• Positive for diphenydramine <50ng/mL
• 0.2 ng/mL for JWH-018 by directed analysis, LC-TOF
Death Investigation Case
• 19 yo male out for a night of partying with friends.
• Smoking marijuana and K2.
• Becomes disoriented.
• Returned home late in evening.
• Found dead in bed the following morning.
• No anatomic cause of death.
• Toxicology (cardiac)
• Delta-9-carboxy THC 13ng/mL
• JWH-018 0.71ng/mL
• Cause of Death – currently undetermined
Conclusions
• Synthetic cannabinoids have great toxicological
significance.
• The range of synthetics is likely to be a moving
target.
• Effect profile very similar to marijuana
• Should be considered in “marijuana” cases where
no THC or metabolites present.
• Blood analysis should target parent drugs <1ng/mL
• Urine analysis must look for multiple points of
identification, as standards are unavailable.
Acknowledgments
• Analysis
• NMS Labs
• Allan Xu
• Matt McMullin
• Lindsay Reinhold
• Donna Warrington
• Webinar
• Amanda Panepinto
• Julie Ruth
• Sherri Kacinko
• Dosing Study
• Bob Welsh
• Tracey Durbin
• DRE’s and subjects
• WSLH
• Amy Miles
www.NMSLabs.com
1.866.522.2216