Barry K Logan PhD, DABFT
Director of Forensic Services
IACP DRE Conference 2010
Pittsburgh PA
New Highs:
Salvia and K2:
Solutions for the DRE
Program
From the Premium Provider of Forensic Services
NMS Labs DRE Offering
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NMS Labs DRE Test Offering
• Blood/Urine
• Scope: Cannabinoids, Cocaine metabolite,
Amphetamines, MDMA, PCP, Barbiturates,
Benzodiazepines, Opiates, Methadone,
Oxycodone, Propoxyphene.
• Confirmations included in price.
• Proactive contact on negative cases.
For informational purposes. NMS Labs, When you need to know, the associated logo and all associated NMS Labs marks are the trademarks of NMS Labs.
© Copyright 2009 NMS Labs. All Rights Reserved. 08/09
Synthetic Cannabinoids
Synthetic drugs that mimic the
effects of cannabis.
Investigational use for appetite,
blood pressure, nausea, etc
First synthesized in the 1970’s
Sprayed onto dried plant leaves,
flowers and stems, and
smoked.
Legal or unregulated in most US
states.
THCHU-210
CP 47,497
Marijuana to Spice
Cannabicyclohexanol
JWH-018 JWH-073
Clemson Series - JWH
JWH-250
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
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 -
Missouri K2 Administration Study
•IRB Human subjects approval obtained from University of
Central Missouri.
•Six subjects smoked K2 Summit, Citron, Standard
•Each contained JWH-018 and 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
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
Synthetic Cannabinoids - Biologicals
• Agilent 6230 TOF Mass Spectrometer
• Agilent 1200 binary UPLC
• ESI source with Nitrogen Jet
• High resolution mode.
• Total run time 10 minutes
• Sensitivity 0.1ng/mL
• Confirmed by LCMS/MS
Analyte Accurate
Mass
JWH-018 341.45272
JWH-073 327.42584
JWH-250 335.44600
Target analytes
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
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
• Positive for JWH-018 by directed analysis, LC-TOF
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
• Salvia; Sal; Magic Mint, Mystic Sage
• Ska Pastora; Sally-D; Diviners Sage; Shepherdess's Herb; ska
Maria Pastora; hojas de Maria; yerba de Maria;
• “Salvia divinorum is a sprawling perennial herb found in the Sierra
Mazatec region of Mexico. Its leaves contain the extremely potent
Salvinorin A. It has a history of buccal use as a divinatory
psychedelic, and has been widely available since the mid 1990s
primarily as a smoked herb. Its effects are considered unpleasant by
many people.”
http://www.erowid.org/plants/salvia/salvia.shtml
Salvia Divinorum
Salvia Divinorum - Use
• Currently sold in head shops, tobacco shops, and online.
• SAMHSA estimates 750K – 1M last year use (2007).
• “Buy Salvia” Google search generates 285,000 hits.
• Available as plants, crushed leaves, extract
• Extracts contain Salvinorin A and B
(mg/g) Sal A Sal B
Leaves 3.2 - 5.0 0.10 – 0.17
Extract 4.1 – 38.9 0.26 – 2.42
Tsujikawa et al, 2008
Salvia Divinorum - Effects
• Following smoking, effects are intense but short lived.
• Psychedelic–like changes in visual perception.
• Mood and somatic changes
• Modified perception of external reality.
• Decreased ability to interact with self or surroundings
Gonzalez, et al, 2006
Salvia– Adverse Effects
Vohra et al, 2009
Clinical Feature Frequency
Altered Mental state 7
Hallucinations 3
Excessive laughter 1
Dizziness 1
Mydriasis 2
Flushing 3
Diaphoresis 1
Tachycardia 2
Palpitations 1
Hypoglycemia 1
•18 Isolated
Salvia
Exposures
Salvia Divinorum - Pharmacodynamics
• C11 Salvinorin A administered to baboons (IV).
• Extremely rapid uptake
• Peak activity achieved in 40 seconds
• Represents 3.3% of dose
• PET kinetics match onset and dissipation in humans
Hooker et al, 2008
Salvia Divinorum - Metabolism
Tsujikawa et al, 2009
NMS Labs - Salvinorin Analysis
• Positive ion electrospray liquid chromatography
tandem mass spectrometry (LC-MS/MS).
• Blood, urine, plasma
• Single step L/L extraction
Analyte MRM MRM RT (min)
Sal A 433/373 433/295 6.67
Sal A-d3 436/376 N/A 6.67
Sal B 389/313 389/295 5.88
Test requirements available at www.nmslabs.com
NMS Labs – Salvinorin B Analysis
• Sal B
• LOQ
– 1ng/mL
• LOD
– ~0.25ng/mL
Salvia Divinorum –Human Study
• Two subjects with prior experience with hallucinogens, smoked 75mg of salvia leaves, for three minutes, ad lib.
• Oral fluid collected at 1 hour post smoking.
• Urine collected at 0-1.5 hrs, and 1.5-9 hrs.
• Sweat patches applied and removed 2 hrs after smoking.
• “Unfortunately… blood collection was not possible… both volunteers experienced intense hallucinations that started about 30 seconds after inhaling peaking at 3-5 minutes, and lasting about 15-20 minutes. During that period blood drawing was considered unsafe, and at the end of the experience the subjects refused blood drawing.”
Picini et al, 2005
Salvinorin A and B – Stability
Days 1 2 9 14 28
Blood NaF 25oC N/Y N/Y N/Y N N
Blood NaF 4oC N/Y N/Y N/Y N N
Blood NaF -20oC Y Y Y Y Y
Plasma NaF 25oC Y Y Y Y N
Plasma NaF 4oC Y Y Y Y Y
Plasma NaF -20oC Y Y Y Y Y
Urine* 25oC Y Y N/Y N N
Urine* 4oC Y Y Y Y Y
Urine* -20oC Y Y Y Y Y
* unpreserved
Acknowledgements
• Analysis
• NMS Labs
• Allan Xu
• Matt McMullin
• Lindsay Reinhold
• Penn. State Police
• Cpl Brian Torkar
• Agilent
• Join Forensic Toxicology on www.LinkedIn.com
• Dosing Study
• UCMO
• Bob Welsh
• Tracey Durbin
• DRE’s and
subjects
• WSLH
• Amy Miles
NMS Labs Salvia and K2 test codes
• Toxicology
• 8070B/S/P/U - Drug Impaired Driving/DRE Toxicology
Panel
• 4150B/S/P/U - Salvinorin A & B
• 7744B/S/P - Special Request for Synthetic Cannabinoids
• Drug Identification
• 7201LI/SL - Controlled Substances/Pharmaceuticals
• 7472SL - Synthetic Cannabinoids – solid
• 7461LI/SL - Salvinorin A identification – liquid/solid