1. Detection of new psychoactive substances (NPS) use across Europe
Sara Castiglioni, Ettore Zuccato, Noelia Salgueiro-González, Emma Gracia Lor
Lisbon, 25th October LX Addiction

2. New Psycoactive Substances (NPS) are
Identification and Assessment of New Psychoactive Substances: A European Network
New Psycoactive Substances (NPS) are
substances not controlled by the international drug control conventions, which may pose public health threat
NPS have similar structure of drugs under international control such as cannabis, cocaine, heroin, MDMA (“ecstasy”), amphetamine

3. Identification and Assessment of New Psychoactive Substances: A European Network
How many?
EMCDDA
EU Early Warning System (EWS) were established to identify these substances in the market in a short time and evaluate health risks
Cooperation with UNODC, Europol, EMA,
WHO
EMCDDA–Europol 2016 Annual Report on the implementation of Council Decision 2005/387/JHA

4. Identification and Assessment of New Psychoactive Substances: A European Network
NPS classes
EMCDDA–Europol 2016 Annual Report on the implementation of Council Decision 2005/387/JHA

5. Few information available
Identification and Assessment of New Psychoactive Substances: A European Network
The route of NPS
EMCDDA– New Psycoactive Substances in Europe
High number of substances
Mostly not controlled
Appear very fast in the market
Used in specific subpopulations
Mixed in unknown cocktails
Potential high toxic effects
Very difficult to assess prevalence of use
Few information available

6. Wastewater – Based Epidemiology (WBE)
Identification and Assessment of New Psychoactive Substances: A European Network
Wastewater – Based Epidemiology (WBE)
Measurement of excretion residues (biomarker) in urban wastewater
Evaluation of consumption or exposure to substances
Almost all substances we consume
are excreted unchanged or as a mixture of metabolites in urine and/faeces
Excreted substances entry in sewer network
Enter sewage treatment plants
Sampling and analysis untreated wastewater

7. WBE Advantages and Challenges for NPS
Identification and Assessment of New Psychoactive Substances: A European Network
WBE Advantages and Challenges for NPS
Ability to provide:
Challenges:
Objective estimates
Limited use
Lots of substances
Low levels in wastewater
Near real-time estimates
Updated information
Human metabolism unknown
Difficult identify biomarkers
Qualitative and quantitative
data
Identification of NPS
Evaluation of use
WBE needs to be tested

8. Preliminary Investigations
Identification and Assessment of New Psychoactive Substances: A European Network
Preliminary Investigations
Europe
González-Mariño et al., ES&T , 2016
Australia
Chen et al., Forensic Science International, 2013

9. Assess the use of New Psycoactive Substances in Europe
Identification and Assessment of New Psychoactive Substances: A European Network
Aim of the study
Identification and Assessment of New Psycoactive Substances in Europe using WBE
Select substances (NPS) to be investigated
Develop a method to screen the presence of NPS (qualitative analyses)
Develop a method to measure NPS (quantitative analyses)
Monitor wastewater samples from different countries
Assess the use of New Psycoactive Substances in Europe
Spatial and Temporal Trends

10. NPS Selection Sources DATABASE 193 substances Criteria
Identification and Assessment of New Psychoactive Substances: A European Network
NPS Selection
Alerts Reports of new substances (December 2013-March 2016)
Sources
DATABASE
193 substances
phenethylamines,
arilcicloexilamine - ketamine analogues,
piperidine,
tryptamine,
synthetic cathinones,
synthetic cannabinoids,
synthetic opioids,
aminorex derivates,
natural psychoactive substances,
benzodiazepines
Alerts (Europe and Italy) + Italy seizure list + Notifications in Italy (intoxications)
(October 2013-March 2016)
UNODC control list
Criteria
Frequency of reporting
Most recently reported

11. Samples investigated COST SCORE Monitoring campaign 2016 (March- May)
Identification and Assessment of New Psychoactive Substances: A European Network
Samples investigated
COST SCORE Monitoring campaign 2016 (March- May)
Italy
Switzerland
Portugal
Slovakia
The Netherlands
Slovenia
Bulgaria
Romania
Poland
Bosnia and Herzegovina
Hungary
Serbia
Ireland
Raw Wastewater Samples
14 European countries
Pooled samples from the weekend

12. Pooled wastewater sample – Solid Phase Extraction
Identification and Assessment of New Psychoactive Substances: A European Network
Samples analyses
Pooled wastewater sample – Solid Phase Extraction
100 mL (pH 7)
100 mL
(pH 2)
Oasis HLB
Oasis MCX
200 µL (10% MeOH)
200 µL (10% MeOH)
The second aliquot was extracted with a Oasis MCX cartridge, which is more selective for basic compounds.
In both cases the eluates were reconstituted in 200 uL, it means that samples were preconcentrated 500 times.
Finally, the extracts were injected in the Orbitrap Qexactive we have.
Each vial was injected 2 times, the first time for a screening and the second one for the confirmation of the suspected compounds.
Orbitrap Q-Exactive
Qualitative Screening
No standards available
API 5500 QqQ
Quantitative Analysis
Standards available

13. Quantitative Analysis - Results
Identification and Assessment of New Psychoactive Substances: A European Network
Quantitative Analysis - Results
Synthetic Cathinones
Mephedrone
Methcathinone
N,N-Dimethylcathinone
Ethcathinone
Methedrone
4-FMC
3,4-DMMC
4-MEC
Buphedrone
Pentedrone
Methylone
Ethylone
Butylone
Pentylone
1-Naphyrone
Naphyrone
MDPV
α -PVP
4-Cl- α -PPP
19 substances searched
9 substances found in wastewater
Phenethylamine
PMA
PMMA
25-B-NBoMe
25-C-NBoMe
25-I-NBoMe
Ketamine analogues
Methoxethamine
Tryptamine
5-MeO-MIPT
7 substances searched
1 substance found in wastewater
Limits of quantification
ng/L

14. Quantitative Analysis - Results
Identification and Assessment of New Psychoactive Substances: A European Network
Quantitative Analysis - Results
Not found in Bosnia Herzegovina and Ucraine

15. Quantitative Analysis - Results
Identification and Assessment of New Psychoactive Substances: A European Network
Quantitative Analysis - Results

16. Qualitative Analysis – Results
Identification and Assessment of New Psychoactive Substances: A European Network
Qualitative Analysis – Results
(193 substances investigated)
Synthetic cathinones
3,4 DmeO-alpha-PVP
Poland, Romania
Tryptamine
α-methyltryptamine
The Netherlands
Aminorex derivate
N-methyaminorex
Slovenia
4,4 DMAR
Ucraine
Phenethylamine
NPDPA
Serbia, Romania
2-methoxyamphetamine
Bosnia Herzegovina
25iP-NBoME
Romania
Synthetic cannabinoids
MDMB-CHMICA
Poland
5-fluoropenthyl-3-pyridinoylindole
Switzerland, Slovakia
Piperidine
2-MeO-diphenidine
The UK
Criteria for Identification:
Accurate Mass
Mass Error < 5 ppm
Presence of Fragments
Standards needed
for confirmation

17. Conclusions WBE was applied successfully to monitor the use of NPS
Identification and Assessment of New Psychoactive Substances: A European Network
Conclusions
WBE was applied successfully to monitor the use of NPS
A qualitative screening was performed for about 200 substances
A quantitative analysis was done for > 25 substances
The levels of NPS were lower than those of the “classical” drugs – lower use ?
Differences among countries were assessed
WBE is a good complementary tool to evaluate the use of NPS

18. Paul Griffith, Liesbeth Vandam, Ana Gallegos
Identification and Assessment of New Psychoactive Substances: A European Network
Acknowledgements
Project Partners
1. Mario Negri Institute (coordinator)
2. University Jaume I
3. Faculdade de Farmacia Universidade de Lisboa
4. Instituto Nacional de Medicina Legal
e Ciencias Forenses
5. Institute of Health Sciences “Egas Monitz”
6. (Norwegian Institute of Water Research)
EMCDDA
Paul Griffith, Liesbeth Vandam, Ana Gallegos
Project Collaborators (who kindly provided samples)
Igor Bodik (Slovakia); Ester Heath (Slovenia); Katarzyna Styszko (Poland); Erik Emke (The Netherlands);
Barbara Kasprzyk-Hordern (UK); Lisa Jones (Ireland); Mariya Skobley and Tanya Bodnarchuk (Ucraine);
Maja Turk Sekulic and Jelena Radonic (Serbia); Andjelka Petkovic (Serbia); Simone Milanolo (Bosnia Herzegovina);
Simion Beldean (Romania); Jelyaz Rangelov and Teodora Todorova (Bulgary); Francesca Pizza (Italy); Francesco Poretti (Switzerland); Miguel Santos (Portugal).
Thanks for your attention!