The EU-project ´SPICE-profiling´ (2015-2017) and other forensic initiatives targeted at NPS

Michael Pütz Federal Criminal Police Office (BKA), Germany Forensic Science Institute

Sarajevo, 03.07.2015 OSCE / UNODC Regional Workshop on „New Trends and Identification Techniques in Detection of NPS and Co-operation among LEA in South East Europe“

2

EU-projects Spice/Spice II plus

DPIP: Drug Prevention and Information Progr.

The EU-project (2010-2012) ‘SPICE and synthetic cannabinoids’: Spice and synthetic cannabinoids: Fast responses by means of forensic, toxicological and socio-scientific analyses with direct impact on prevention measures. Legal medicine, Universities (Toxicological and Drug Research), Police/Forensics, Drug Prevention and Monitoring Institutions Freiburg, Wien, Helsinki, Frankfurt, Wiesbaden, Warschau, Bern

Continuation (2013-2015) in the EU-project ´SPICE II plus´: Extension to substance classes of stimulant NPS (a. o. cathinone and piperazine derivatives), even stronger focus on prevention aspects and intensified co-operation with drug monitoring centres (DBDD, EMCDDA). Legal medicine, Universities (Toxicological and Drug Research), Police/Forensics, Drug Prevention and Monitoring Institutions Freiburg, Wien, Helsinki, Frankfurt, Wiesbaden, Bern, München, Lissabon

Prevention measures (drug supply/demand/harm reduction)

3

EU-projects Spice/Spice II plus

DPIP: Drug Prevention and Information Progr.

Product monitoring of ‘Spice’, ´bath salt´ and other NPS products

Identification of new compounds

Build-up of mass spectrometric libraries (EI-MS and ESI-MS/MS)

Assessment of toxicological properties and risk profiles

Development of bioanalytical methods (metabolism!)

Rapid detection, ‘profiling’ concepts, quantitation in drug material

Social research (prevalence, patterns of use)

International conference on ‘Spice’ prevention issues

‘Spice’ information and prevention website, brochure and flyers:

http://legal-high-inhaltsstoffe.de/

Results in positive serum samples (n=734) 05/2010 – 03/2012 (selection of compounds)

0%

20%

40%

60%

80%

100%

JWH-250 JWH-081, JWH-122 JWH-203, JWH-210, RCS-4 Scheduled to be controlled

Rel

ativ

e fr

eque

ncy

of o

ccur

renc

e

JWH-081 JWH-122 JWH-250 JWH-210 RCS-4 JWH-203 AM-2201 JWH-019 JWH-307 AM-1220

Method: Journal of Mass Spectrometry, Dresen et al. 2010, Kneisel et al. 2012

Prevalence of synthetic cannabinoids

67

9

7

3

12

1

Lebenszeit-Prävalenz 30-Tages-Prävalenz

Spice/ "Räuchermischungen"

2008 2009 2010 2011

Life time prevalence 30-day prevalence

‘Spice products’

2012: 7-8% of 15-18-year-old pupils in Frankfurt, Germany consumed SPICE-products

Prevalence of synthetic cannabinoids

- 2012: 7-8% of 15-18-year-old pupils in Frankfurt, Germany consumed SPICE-products

- 8 % of urine samples from ‘fitness to drive’ testing (negatively tested for THC-COOH) were positive for SC’s

- Positivity rates in forensic-psychiatric hospitals: up to > 80 % (trying to escape drug testing)

- Finland: 3 % of DRUID drivers had synthetic cannabinoids in their blood in addition to other drugs

- In some countries less problems with SC’s (why?)

77

62

61

57

3734

33

13

115

'High'

Curiosity

Legal availability

Relaxation

Variety

Non-detectability

Other drugs (temporarily) not available

(low) Price

Because friends comsume, too

Because I had problems

"wichtig/ sehr wichtig"

Goethe-Universität Frankfurt am Main Institut für Sozialpädagogik und Erwachsenenbildung

Online-survey: Motives for consumption (%)

(‚Legal highs‘)

‘Important / very important’

Brochures and Flyers directed to consumers and professional

http://legal-high-inhaltsstoffe.de/

Procedures for rapid detection, identifi-cation and quantification of seized NPS

- Ion Mobility Spectrometry (IMS)

- FT-IR and Raman spectroscopy

- Ambient MS (DESI–MS)

- Quantification (GC-FID, UHPLC-DAD, LC-MS)

► UNODC manual STNAR48 (released 06/2013, Auwärter/Pütz)

GTFCH Symposium Mosbach 10 Comparison of rapid detecting optical techniques for the identification of New Psychoactive Substances (NPS) in ‘Legal Highs’ and ‘Research Chemicals’

Rapid Detection Techniques Application fields & Challenges

Police Street- and traffic controls

Seizures and big events

Customs Suspicious material/samples

Forensics Clandestine drug labs

Explosives/Disarming

Short analysis time

Simple & safe handling

Portability

Reliable detection/analysis of substances/mixtures

GTFCH Symposium Mosbach 11 Comparison of rapid detecting optical techniques for the identification of New Psychoactive Substances (NPS) in ‘Legal Highs’ and ‘Research Chemicals’

Presumptive Drug Tests (Color reaction)

IMS (Ionmobility-spectrometry)

Ambient MS-Techniques (DESI-MS, DART)

Spectroscopic Techniques (FT-Infrared[IR] & Raman) Specific/Structural substance

information (fingerprint region)

Non-invasive

Quantification possible

Less sample preparation

Detection Limit ~ 5 – 10 %

Library dependent

Rapid Detection Techniques

GTFCH Symposium Mosbach 12 Comparison of rapid detecting optical techniques for the identification of New Psychoactive Substances (NPS) in ‘Legal Highs’ and ‘Research Chemicals’

Infrared vs. Raman Spectroscopy Pros & Cons

Source: HazMat ID Product Sheet, Smiths Detection (left) Progeny Product Sheet, Rigaku (right)

GTFCH Symposium Mosbach 13 Comparison of rapid detecting optical techniques for the identification of New Psychoactive Substances (NPS) in ‘Legal Highs’ and ‘Research Chemicals’

Infrared vs. Raman Spectroscopy Pros & Cons

FT IR Raman Basis Absorption of light by

functional groups/bonds Inelastic scattering of light by

molecules

Signal intensity High Low

Preheating necessary

Yes No

Spectrum Substraction

Yes No

Analysis through packaging

No Yes

Disturbed by Water Fluorescence (@785 nm)

Raman 785 nm vs. 1064 nm Synthetic Cannabinoids

GTFCH Symposium Mosbach 14 Comparison of rapid detecting optical techniques for the identification of New Psychoactive Substances (NPS) in ‘Legal Highs’ and ‘Research Chemicals’

0

500

1000

1500

2000

2500

3000

400 600 800 1000 1200 1400 1600

Inte

nsit

y

Ramanshift [cm-1]

XLR-11

1064 nm 785 nm

Raman Handheld-Systems (785 nm/1064 nm): Metrohm

N

O

F

CH3 CH3

CH3CH3

0

1000

2000

3000

4000

5000

6000

7000

8000

9000

400 600 800 1000 1200 1400 1600

Inte

nsit

y

Ramanshift [cm-1]

STS-135

1064 nm 785 nm

NH

N

O

F

GTFCH Symposium Mosbach 15 Comparison of rapid detecting optical techniques for the identification of New Psychoactive Substances (NPS) in ‘Legal Highs’ and ‘Research Chemicals’

0

500

1000

1500

2000

2500

400 600 800 1000 1200 1400 1600

Inte

nsit

y

Ramanshift [cm-1]

1064 nm

STS-135

0

200

400

600

800

1000

1200

400 600 800 1000 1200 1400 1600

Inte

nsit

y

Ramanshift [cm-1]

785 nm

XLR-11

Raman 785 nm vs. 1064 nm Synthetic Cannabinoids

Raman Handheld-Systems (785 nm/1064 nm): Metrohm

Enough Information for Identification

Not enough Information for Identification

GTFCH Symposium Mosbach 16 Comparison of rapid detecting optical techniques for the identification of New Psychoactive Substances (NPS) in ‘Legal Highs’ and ‘Research Chemicals’

0.01

0.02

0.03

0.04

0.05

0.06

0.07

0.08

0.09

1150 1250 1350 1450 1550 1650 1750Wavenumber [cm-1]

Herbal Blend 'R&B'

R&B Acetone Extract R&B untreated

Library Hits: R&B Untreated THC R&B Acetone extract JWH-210

Picture:http://i1.cpcache.com/product_zoom/1534831882/samsung_galaxy_s5_case.jpg?height=250&width=250&padToSquare=true

Herbal Blend + 50 µL acetone

Extraction

Transfer on ATR spot Result

Measure after drying

FT-IR Synthetic Cannabinoids – Herbal Blends

Pre-treatment: Acetone Extraction

FT-IR Mobile-System ‚HazMat ID®‘: Smiths Detection

GTFCH Symposium Mosbach 17 Comparison of rapid detecting optical techniques for the identification of New Psychoactive Substances (NPS) in ‘Legal Highs’ and ‘Research Chemicals’

-0.05

0

0.05

0.1

0.15

0.2

0.25

650 1150 1650 2150 2650 3150 3650Wavenumber cm-1

Bath Salt 'Mojo'

Mojo Mojo Residual

-0.04-0.02

00.020.040.060.080.1

0.120.140.160.18

650 850 1050 1250 1450 1650Wavenumber cm-1

Bath Salt 'Mojo' – Residual

MDPV Mojo Residual

FT-IR Synthetic Cathinones – Bath Salts

1. Library Hit: Lidocaine 2. Library Hit: MDPV

Spectrum Substraction

FT-IR Mobile-System ‚HazMat ID®‘: Smiths Detection

Conclusion

FT-IR Research Chemicals

Herbal Blends (pretreatment)

Bath Salts (concentration dep.)

× Liquids

× Packed Samples

GTFCH Symposium Mosbach 18 Comparison of rapid detecting optical techniques for the identification of New Psychoactive Substances (NPS) in ‘Legal Highs’ and ‘Research Chemicals’

Raman @1064 nm Research Chemicals

Bath Salts (concentration dep.)

Liquids (concentration dep.)

Packed Samples

× Herbal Blends

Combination of FT-IR and Raman (@1064 nm) spectroscopy can deliver reliable results with structural information in short time without lab environment

User must be trained Limits of both techniques must be known

15.04.2011 19 Identification of synthetic cannabimimetic substances in herbal mixtures

Rapid analysis of NPS by IMS

Ion Mobility Spectrometer DE-tector, Bruker Daltonik

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

10

20

30

40

50

Inte

nsitä

t

Driftzeit in ms

Coca

ine

Drift time window synthetic cannabinoids

Calib

rant

Drift time window classic drugs

20 SPICE II

Rapid detection of cathinone derivatives in bath salts by IMS

3D-plasmagram of bath salt sample ‚Mojo‘ identifes MDPV and lidocaine

“SPICE-Profiling: Assessment of the origin, supply chain and market development of NPS by forensic analysis of the compo-sition and of the chemical, metabolic and isotopic signatures”

ISEC 2013, priority EPSD, JUST/2013/ISEC/DRUGS/AG/ISEC/4000006421 Co-ordinator: BKA, Forensic Science Institute, Wiesbaden Funded Institute of Legal Medicine, Freiburg, Germany partners: INPS, French Police Forensic Science Institute, Lyon Associated Finnish National Customs Laboratory, Helsinki partners: LKA RP LKA SH LKA BY LKA HE Duration: 01.04.2015 – 31.03.2017 Budget: ~700 TEUR (90 % funded)

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EU-project „SPICE-profiling“

ISEC: Fight Against and Prevention of Crime

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„SPICE-profiling“: core activities

Workstream 1: Product monitoring of products containing NPS by qualitative and quantitative analysis of products composition (internet test purchases of products and pure substances, police and customs seizures).

Workstream 2: Method development for clinical and forensic analysis of human biosamples and application to forensic case work and clinical emergency cases.

Workstream 3: - Assessment of suitable synthesis impurities in unadulterated NPS samples by chemical analysis via GC-MS and UHPLC-MS and conducting controlled syntheses of synthetic cannabinoids. - Development of flexible impurity profiling procedures for synthetic cannabinoids and synthetic cathinones via GC-MS, creation of a representative database and multivariate analysis of impurity profiling and isotopic profiling data.

Workstream 4: Development, validation and harmonization of extraction techniques and of methods to determine isotope ratios using EA-IRMS and GC-IRMS-MS of synthetic cannabimimetic substances and application to establish links between different seizures and to link seizures to clandestine producers.

„Research Chemicals“ – where do they come from?

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Clandestine production of NPS in China

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Pictures taken from a YouTube video, showing the production of JWHs in a clandestine lab

titaniumwf.com Manufacture JWH. Sale of technology of synthesis JWH, MDPV 2011 [cited 2014 08 June]; Available from: www.youtube.com/watch?v=eho28ALAD-c

Scheme for profiling analysis

GC/MS

Combined impurities

Isolation

Flash column chromatography

Pure substance

Reference material

HS-GC, ICP-MS

Controlled synthesis

Identification, chrom. Profiling

EA-IRMS, GC-IRMS

Multivariate data analysis

Sample links, information on origin

Flash chromatography for isolation of main active NPS substances and combined fractions of impurities

Sepracore flash chromatography Full automated fraction control via UV/vis detector Two pumps for up to 50 bar Ready-made column cartridges and self-packed glass columns

Example: XLR-11 synthesis impurities

NH

O

NH

O

NUR-144

1. EtMgBrTHF2. ZnCl2Et2O

O

3. CH2Cl2

Cl

1. NaHDMF

2. DMFBr

O

NH

m/z 242C16H20NO+

O

N

F

O

+ H+

m/z 454C29H41FNO2

+

+ H+

N

F

F

+ H+

m/z 294C18H26F2N+

O

N

O

+ H+

m/z 366C24H32NO2

+

Published synthesis of UR-144 Frost, J.M., et al., Indol-3-ylcycloalkyl Ketones: Effects of N1 Substituted Indole Side Chain Variations on CB2 Cannabinoid Receptor Activity. Journal of Medicinal Chemistry, 2009. 53(1): p. 295-315.

XLR-11 impurities tentatively identified in extract of Manga XXL via UHPLC-MS n

Impurities hint to comparable synthesis route for XLR-11 starting from indole (high amounts of impurities)

Practical application of chromatographic impurity profiling in clandestine labs

distillation residue from white cannister (6,6 kg, 10 % amph.)

sample from distillation flask (res.,10 % amph.)

strong link (both samples ori- ginating from one production batch)

22.03.2011

29 Das Bundeskriminalamt - Zentralstelle der deutschen Kriminalpolizei

Basics of stable isotope ratio analysis

Isotope effects

1H1HO evaporates easier than 2H1HO

⇒ water steam is depleted in heavy hydrogen isotopes compared to lake water

precipitation is enriched in heavy hydrogen isotopes compared to water

steam

Example: water cycle

H2O (l) H2O (g)

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EA-IRMS batch-to-batch comparison of 5F-PB-22 extracted from Spice-products

7.00

7.20

7.40

7.60

7.80

8.00

8.20

8.40

8.60

8.80

9.00

-26.10 -25.90 -25.70 -25.50 -25.30 -25.10 -24.90 -24.70 -24.50

δ15 N

AIR[

‰]

δ13CV-PDB[‰]

Made bin Belgium

Jamaican Gold Extreme

Bonsai Winter Boost

Love

R&B

Bubblegum

Yama

Blaze

Manga Extreme

Bonsai Citrus

Bonsai Summer Boost

Saarbrücken

Bonzai Summer Boost (2)

Jamaican Gold Supreme

VIP

Recent activity: MDMB-CHMICA

Large seizure of 40 Kg pure MDMB-CHMICA by LUX customs (December 2014)

Ca. 50 Mio. single doses

From Shanghai, Destination: Madrid

40 bags à 1 Kg

Samples of all 40 bags kindly provided to BKA (one bigger sample to distribute to LKÄ/BWZ)

First information: Batch production size!

Other ongoing EU initiatives related to NPS

EU-project RESPONSE (ISEC, 2015-2017) CLEN network (EU forensic customs labs, JRC, EMCDDA, DG TAXUD)

COLLECT, ANALYSE, ORGANIZE, EVALUATE, SHARE – A RESPONSE TO CHALLENGES IN FORENSIC DRUGS ANALYSES

project objectives and activities

Sonja Klemenc (RESPONSE project manager) (sonja.klemenc@policija.si and ssklemenc@gmail.com )

MNZ GPU, National Forensic Laboratory, Vodovodna 95, 1000 Ljubljana, Slovenia

ENFSI- DWG meeting, 5th-8th May, Dublin 2015

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PARTNERS

ENFSI- DWG meeting, 5th-8th May, Dublin 2015 39

Project was developed and will be implemented in close cooperation with ENFSI WORKING GROUP DRUGS Grant agreement number: JUST/2013/ISEC/DRUGS/AG/6413

Duration: from 5th January 2015 until 4th January 2017 Total project budget : 748.212,91 € (EU contribution: 90%, R Slovenia 10%)

Coordinator: Ministry of the Interior of the Republic Of Slovenia, Police – NATIONAL FORENSIC LABORATORY, Slovenia

Partners (co-beneficiaries): Ministry of the Interior, National Forensic Institute (INPS), France Hungarian Institute for Forensic Sciences (HIFS), Hungary National Institute of Criminalistics and Criminology (INCC), Belgium Institute for Forensic Medicine, Aarhus (UNI Aarhus), Denmark University of Copenhagen, Faculty of Health (UNI CPHG), Department of Forensic Medicine, Denmark Ministry of the Interior of the Republic of Croatia, Forensic Science Centre (MUP RH), Croatia University of Ljubljana, Faculty of Chemistry and Chemical Technology (FKKT), Slovenia

Associate partners: European Monitoring Centre for Drugs and Drug Addiction (EMCDDA), Portugal Forensic Science Lab, Dublin (FSL), R Ireland Forensic Science and Toxicology Lab, State General Laboratory, Ministry of Health (FSTL), Cyprus Judiciary Police, Scientific Police Laboratory (SPL), Portugal National Investigation Service Norway (KRIPOS), Norway National Institute for Public Health (NIJZ), Slovenia University of Ljubljana, Faculty of Medicine, Department of Toxicology (UNI LJ FM DT), Slovenia Association DrogArt (DrogArt) NGO, Slovenia

Project impact

ENFSI- DWG meeting, 5th-8th May, Dublin 2015 40

Forensic information is the key factor for the: Strategic Operational Evidential Judicial purposes

And influences the: Drug supply reduction Planning Risk assessment activities Coordination of activities at all EU levels and wider

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Thanks to the co-workers and for your attention!