k2 and the synthetic cannabinoids: pharmacology, effects and chemical analysis
DESCRIPTION
Presented September 16, 2010 by Dr. Barry K. Logan, PhD, DABFT, NMS Labs National Director of Forensic Services NMS Labs has responded to the recent explosive growth in popularity of synthetic cannabinoid agonists in so-called “herbal blends” by developing new tests for the active chemicals in botanical material, and most recently for the parent drugs and their metabolites in blood and urine. This presentation describes the history and origin of the chemicals of concern, the composition of the various commercial products containing them, their known pharmacology, and the documented effects on drivers, and human test subjects. We also review the adverse effects that have resulted in hospitalization, and even allegedly in deaths. This presentation describes the challenges around providing a chemical test for these new drugs, information on their stability in biological fluids, and the validation of quantitative methods for their determination.TRANSCRIPT
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Barry K Logan PhD, DABFTDirector of Forensic Services
NMS Labs online SeminarSeptember 16, 2010
K2 and the Synthetic Cannabinoids: Pharmacology,
Effects, and Chemical Analysis
From the Premium Provider of Forensic Services
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K2 Phenomenon
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K2 Phenomenon
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K2 Phenomenon
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K2 Phenomenon
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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.
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MarijuanaMost 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.
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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.
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K2 Phenomenon
Kansas, Georgia,
Kentucky, Alabama,
Tennessee, Louisiana,Missouri, Hawaii,
Arkansas,Illinois
Mississippi
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JWH-018 JWH-073
Clemson Series - JWH
JWH-250
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THC HU-210
CP 47,497
Marijuana to Spice
WIN 55,212
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Cannabinoid Mechanism of ActionCerebral Cortex
Higher Cognitive FunctionBasal GangliaCerebellum
MovementHippocampus
Learning, Memory, StressHypothalamus
AppetiteSpinal Chord
Pain, Peripheral sensationMedulla
Nausea, Vomiting
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Herbals - Botanical Substrate
Glycyrrhiza glabra(Cultivated Liquorice)
Astragalus membranaceus(Milk Vetch)
Verbascum thapsus(Common Mullein)
(Uchiyama et al, 2010)
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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
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Synthetic Cannabinoids – Drug ID
JWH-018 JWH-073 JWH-200HU-210/11JWH-019JWH-200
WIN 55,212 JWH-015JWH-133 JWH-250 CP55,940
CP 47,497 (n=7) CP 47,497 (n=8)
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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 - 14K2 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 -
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Synthetic Cannabinoid Effects
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Synthetic Cannabinoid Effects
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Synthetic Cannabinoid Metabolism
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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
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Synthetic Cannabinoid Disposition
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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)
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NMS Labs Special ProjectsMatrix Analytes
BloodSerumOral 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 – glucuronidatedRing dihydroxylated metabolites – glucuronidatedDesmethyl ring hydroxylated metabolites - glucuronidated
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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.
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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
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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
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K2 use and the DRE Matrix
Drug Symptom Matrix
CNS
Depressant InhalantsDissociative
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
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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
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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
7.376.747.866.17
BM 2 hr Post dose blood
2 HR POST DOSE BLODD
IS
JWH-018
JWH-073
HO JWH-073
HO JWH-018
Printing Date: Wednesday, September 15, 2010 Page 1 of 1 Acq. File: ED01_080610K2_AX.wiff
Sample Name: BM 2 hr Blood #2 not hydrolyzed Sample Comment: Project: Production
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Missouri K2 Administration Study
Subject BM
Smoked “K2 Citron”
10mg/g JWH-018/073
0.3g in water pipe
3 inhalations over 30 minutes
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Synthetic Cannabinoid Disposition
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Synthetic Cannabinoid Disposition
* *
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NMS Labs Urine JWH-018 and -073L/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
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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
4.0e64.2e6
Inte
ns
ity, c
ps
6.80
7.33
6.003.47
JWH-018/073 Human Urinary Metabolites
IS
HO
HOCOOH
HO-N-dealkylation2HO
3HO
Printing Date: Wednesday, September 15, 2010 Page 1 of 1 Acq. File: ED01_090310K2_AX.wiff
Sample Name: Hydrolyzed 19-20-21 mix 1 Sample Comment: Project: Production
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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-
073Mono-OH Di-OH Tri-OH Glucuronide
s
Pre-dose
X X X X X
1:15 X √ √ X √
2:07 X √ √ √ √
2:40 X √ √ √ √
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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
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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
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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
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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
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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.
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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