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

K2 Phenomenon

K2 Phenomenon

K2 Phenomenon

K2 Phenomenon

K2 Phenomenon

K2 Phenomenon

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.

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.

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 ActionCerebral Cortex

Higher Cognitive FunctionBasal GangliaCerebellum

MovementHippocampus

Learning, Memory, StressHypothalamus

AppetiteSpinal Chord

Pain, Peripheral sensationMedulla

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-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)

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 -

Synthetic Cannabinoid Effects

Synthetic Cannabinoid Effects

Synthetic Cannabinoid Metabolism

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 Cannabinoid Disposition

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 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

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 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

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

7.376.747.866.17

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

Missouri K2 Administration Study

Subject BM

Smoked “K2 Citron”

10mg/g JWH-018/073

0.3g in water pipe

3 inhalations over 30 minutes

Synthetic Cannabinoid Disposition

Synthetic Cannabinoid Disposition

* *

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

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

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 √ √ √ √

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

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