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K2 and the Synthetic Cannabinoids: Pharmacology, Effects, and Chemical Analysis Barry K Logan PhD, DABFT Director of Forensic Services NMS Labs online Seminar September 16, 2010

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.

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

Clemson Series - JWH

JWH-018

JWH-073

JWH-250

Marijuana to Spice

THC

CP 47,497

HU-210

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

Verbascum thapsus (Common Mullein)

(Uchiyama et al, 2010)

Astragalus membranaceus (Milk Vetch)

Glycyrrhiza glabra (Cultivated Liquorice)

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 K2 Blonde K2 Standard K2 Citron K2* (Unknown) K2 Summit K2 Blue K2 Pink K2 Latte K2 Mint K2 Silver Spike Gold Spike Maxx Spike Diamond Spike Silver Space

JWH-018 (mg/g) 12 9 10 11 15 11 16 19 8 20 17 17 9 10

JWH-073 (mg/g) 13 9 10 9 0.28 0.30 11 0.07 16 -

Herbal blends*

2.0 – 35.9

-

(* Uchiyama et al, 2010)

CP47,497 JWH-250 (n=8) (mg/g) (mg/g) 6 14 16 19 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 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

Yes

Yes

Yes

No

No

Vertical Nystagmus

Present *

Present*

Present

No

Lack of Convergence

Present

Present

Present

Pupil Size

Normal

Normal

Reaction to Light

Slow

Pulse Rate Blood Pressure

Horizontal nystagmus

Body Temperature

Dissociative CNS Narcotic Drugs Stimulants Hallucinogens analgesics

Cannabis

K2

No

No

No

No

No

No

No

No

No

No

Present

Present

Normal

Dilated

Dilated

Constricted

Dilated*

Normal

Slow

Normal

Slow

Normal

Little to none

Normal

Normal

Down

Up

Up

Up

Up

Down

Up

Up

Down

Up/Down

Up

Up

Up

Down

Up

Up

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 BM 2 hr Post dose blood

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

XIC of +MRM (34 pairs): 358.2/155.0 amu from Sample 3 (BM 2 hr Blood #...

Max. 2.6e5 cps.

4.0e6

In t e n s ity , c p s

3.5e6

2 HR POST DOSE BLODD

3.0e6

IS

2.5e6

HO JWH-073

2.0e6 1.5e6

JWH-018

1.0e6

JWH-073 HO JWH-018

5.0e5

6.74 0.0

1

Sample Name: BM 2 hr Blood #2 not hydrolyzed Sample Number: N/A Batch Name: K2 080610.dab

2

3

4

Sample Comment: Collected by: NMS815\labman Sample ID: 3

5 Time, min

6.17 6

7.37

7 Project: Production Method Name: N/A Results Name: N/A

7.86 8

9

Missouri K2 Administration Study 155

6

150 JWH-018

145

Concentration (ng/mL)

JWH-073 Systolic BP

4

140 135

3

130 125

2

120 1

115 0

0:00

1:12

2:24

DT (hours)

110 3:36

Systolic Blood Pressure (mm Hg)

5

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 -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 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

XIC of +MRM (35 pairs): 358.2/155.0 amu from Sample 2 (Hydrolyzed 19-2... 4.2e6 4.0e6

Max. 4.2e6 cps.

6.80

JWH-018/073 Human Urinary Metabolites

In t e n s ity , c p s

3.5e6 3.0e6

HO 2.5e6 2.0e6 1.5e6 1.0e6

2HO HO-N-dealkylation

5.0e5

Sample Name: Hydrolyzed 19-20-21 mix 1 Sample Number: N/A Batch Name: K2 090310.dab

2

HO

IS

6.00

3.47 0.0

7.33

COOH 3HO 4 Sample Comment: Collected by: NMS815\labman Sample ID: 3

6 Time, min

8

10 Project: Production Method Name: N/A Results Name: N/A

12

Missouri K2 Administration Study Subject BM - Urine Time

JWH-018

Mono-OH

Di-OH

Tri-OH

Glucuronides

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

√

√

√

√

Pre-dose

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