1. Identification and Assessment of New Psychoactive Substances: A European Network
Identification and assessment of new psychoactive substances: a European network - NPS-Euronet (Home/2014/JDRU/AG/DRUG/7086)
Félix Hernández
Research Institute for Pesticides and Water (IUPA)
Laboratory of Pesticide Residue Analysis (LARP)
Universitat Jaume I, Castellón, Spain
Lisbon, 28th September 2016

2. Urine samples Provided by Origin Location Sample # INMLFC
Identification and Assessment of New Psychoactive Substances: A European Network
Urine samples
Provided by
Origin
Location
Sample #
INMLFC
(Mario Joao Dias)
Emergency department of central hospitals
Lisbon, Porto and Coimbra (Portugal)
Total 90
(5 i – 85 p)
TICTAC
(Trevor Shine)
Creamfields Festival 2015*
Daresbury (England)
Total 31
(31 p)
Juvenile detention center
Picassent (Valencia, Spain)
Total 227 i
Castellón de la Plana (Spain)
Total 23 i
Burjassot (Valencia, Spain)
Total 99 i
Valencia (Spain)
Total 108 i
Total 26 i
1st Batch processed
(19 samples:
5 i – 14 p)
* Confidential information
i = individual sample
p = pooled sample
Lisbon, 28th September 2016

3. (β-glucuronidase from E.Coli)
Identification and Assessment of New Psychoactive Substances: A European Network
Urine samples
Hydrolysis
Without Hydrolysis
1,25 mL urine sample
1 mL urine sample
Deglucuronidation
(β-glucuronidase from E.Coli)
pH 7, 55ºC for 1 hour
Centrifuge
+ 1ml Milli-Q H2O
(Dilution (2x))
Centrifuge
20uL extract
20uL
LC-QTOF MS
LC-QTOF MS
Lisbon, 28th September 2016

4. Wastewater samples Provided by Origin Location Sample # FFUL Mutela
Identification and Assessment of New Psychoactive Substances: A European Network
Wastewater samples
Provided by
Origin
Location
Sample #
FFUL
(Alvaro Lopes)
Mutela
Almada (Portugal)
1 p 3 days
Alcântara
Lisbon (Portugal)
USC
(José Benito Quintana)
Santiago de Compostela
Santiago (Spain)
CSIC
(Cristina Postigo)
El Prat
Barcelona (Spain)
2 i
UJI
Castellón
Castellón (Spain)
Molina de Segura
Murcia (Spain) UA
(Alexander van Nuijs)
Tomorrowland Festival 2015
Belgium *
i = individual sample
p = pooled sample
* To be submitted by Alexander van Nuijs, not sure how many samples
Lisbon, 28th September 2016

5. Wastewater samples Screening Target(1) LC-QTOF MS LC-MS/MS Compounds
Identification and Assessment of New Psychoactive Substances: A European Network
Wastewater samples
Screening
Target(1)
Compounds
N-ethylcathinone
MDPV
Methylone
Butylone
Methedrone
Mephedrone
Naphydrone
25-C-NBOMe
25-I-NBOMe
25-B-NBOMe
25 mL filtered sample
+75mL MilliQ H2O (dil x4)
100 mL centrifuged sample (pH 2)
+ Internal standard
Oasis MCX
(cationic exchange cartridge)
Elution: 5ml MeOH (2% NH3)
Oasis HLB
(generic cartridge)
Elution: 5ml MeOH
LC-QTOF MS
LC-MS/MS
20uL extract
3uL extract(2)
Based on Bade et al, Chemosphere (submitted)
Extract will also be injected for screening by LC-QTOF MS
Lisbon, 28th September 2016

6. Urine samples, Norwegian festivals
Identification and Assessment of New Psychoactive Substances: A European Network
Urine samples, Norwegian festivals
Origin
Music Style
Location
Date
Sample #
Øya Festival* **
Alternative, Electronica, hip hop, indie and rock
Oslo
11-13 August 2016
3 Pooled urine
Pstereo Festival*
Electronica, pop and rock
Trondheim
19 August 2016
1 Pooled urine
Findings Festival*
Electro house and dance pop
20 August 2016
* Confidential information
** Collected by Francesco (MN). These samples will be analyzed also by MN.
Lisbon, 28th September 2016

7. Urine samples, Norwegian festivals
Photos + fechas + muestras + festival (tipo) + sample prep in Norway (SPE) is similar to wastewater (previous slide) only sample volume was modified)
Identification and Assessment of New Psychoactive Substances: A European Network
Urine samples, Norwegian festivals
Pooled urine samples
Treated as urine
Hydrolysis
Dilution
Treated as WW
SPE Oasis HLB
SPE Oasis MCX
Lisbon, 28th September 2016

8. Priority list of NPS Information included:
Identification and Assessment of New Psychoactive Substances: A European Network
Priority list of NPS
Compiled by Mario Negri and University Jaume I
(To be updated)
Information included:
Compound name
IUPAC name
Group (cathinone, cannabinoid, etc)
Communication
Date
CAS number
Formula
Place
Present in the first list (Nov 2013)
Additional information
Under control (Included in List DPR 309/90)
LC Fragments
GC Fragments
Instrument
References
Info metabolism
References metabolites
Standard available (Mario Negri /IUPA)
Lisbon, 28th September 2016

9. Priority list of NPS (MN + UJI)
Identification and Assessment of New Psychoactive Substances: A European Network
Priority list of NPS (MN + UJI)
Group
Number #
Phenethylamine 37
Arilcicloexilamine - ketamine anal. 3
Piperidine 6
Tryptamine 7
Cathinone 53
Synthetic cannabinoid 71
Synthetic oppioid 9
Aminorex derivate
Natural psychoactive substance 4
Benzodiazepine
Total # compounds
197
Compounds:
Lisbon, 28th September 2016

10. UNIFI – software package of Waters
Identification and Assessment of New Psychoactive Substances: A European Network
UNIFI – software package of Waters
Good mass error
Low RT deviation
5 frag. found by software
Bijlsma, L. et al. J. Mass Spectrom. 2011, 46,
Identification
False positive
m/z
Lisbon, 28th September 2016

11. Lisbon Urines, Positives, Ketamine
Identification and Assessment of New Psychoactive Substances: A European Network
Lisbon Urines, Positives, Ketamine
Also major metabolites found (Tentatively identified):
Norketamine
Dehydronorketamine
Hydroxynorketamine
m/z
C7H6Cl (-0.7 mDa)
m/z
C11H12Cl (0.4 mDa)
m/z
C12H12ClO (0.3mDa)
Lisbon, 28th September 2016

12. Lisbon Urines, Tentative Identification, Mexedrone
Identification and Assessment of New Psychoactive Substances: A European Network
Lisbon Urines, Tentative Identification, Mexedrone
m/z
C10H13N (0.01 mDa)
m/z
C12H16NO (-0.2 mDa)
Lisbon, 28th September 2016

13. Tentatively Identified Other compounds detected:
Identification and Assessment of New Psychoactive Substances: A European Network
Results NPS, Lisbon Urines (Samples 1-19)
Identified
Tentatively Identified
Other compounds detected:
Ketamine
Mexedrone
MDMA
Norketamine
Cocaine
Dehydronorketamine
Benzoylecgonine
Hydroxynorketamine
11-nor-9-carboxy-Δ9-THC (THC-COOH)
Procaine
Desmethylprodine
2-MeO-Ketamine
O-Desmethyltramadol
JWH-398
MDCPM
5-Me-MDA
2C-YN (phenethylamine analog)
Lisbon, 28th September 2016

14. Further actions Confirm tentative identifications
Identification and Assessment of New Psychoactive Substances: A European Network
Further actions
Confirm tentative identifications
Harmonize criteria for tentative identifications
Urgent need of reference standards acquisition
Complete analysis of urine and wastewater samples
Update the priority list of NPS with compounds identified in the samples
Lisbon, 28th September 2016

15. Output – Conferences Wastewater based epidemiology: recent advances
Identification and Assessment of New Psychoactive Substances: A European Network
Output – Conferences
Wastewater based epidemiology: recent advances
Lubertus Bijlsma, Pim de Voogt, Sara Castiglioni, Adrian Covaci, Erik Emke, Félix Hernández, Barbara Kasprzyk-Hordern, Christoph Ort, Malcolm Reid, Kevin V. Thomas, Alexander L.N. van Nuijs
Oral presentation
Lisbon, 28th September 2016

16. Potential of HRMS (LC-QTOF MS) in wastewater drug testing
Identification and Assessment of New Psychoactive Substances: A European Network
Output – Conferences
Potential of HRMS (LC-QTOF MS) in wastewater drug testing
Lubertus Bijlsma
Oral presentation
Garching / Munich, November 2016
Lisbon, 28th September 2016

17. Science of the Total Environment 565 (2016) 984–993
Identification and Assessment of New Psychoactive Substances: A European Network
Output – scientific peer-reviewed articles
Science of the Total Environment 565 (2016) 984–993
Lubertus Bijlsma acknowledges NPS-Euronet for his post-doctoral fellowship
Lisbon, 28th September 2016

18. Output – scientific peer-reviewed articles
Identification and Assessment of New Psychoactive Substances: A European Network
Output – scientific peer-reviewed articles
Ultra-high performance liquid chromatography-tandem mass spectrometry determination of synthetic cathinones and phenethylamines in influent wastewater
Richard Bade, Lubertus Bijlsma, Juan V. Sancho, J. A. Baz-Lomba, Sara Castiglioni, Erika Castrignanò, Ana Causanilles, Emma Gracia-Lor, Barbara Kasprzyk-Hordern, Juliet Kinyua, Ann-Kathrin McCall, Alexander L.N. van Nuijs, Christoph Ort, Benedek G. Plósz, Pedram Ramin, Nikolaos I. Rousis, Yeonsuk Ryu, Kevin V. Thomas, Pim de Voogt, Ettore Zuccato, Felix Hernandez
Submitted Chemosphere +
Lubertus Bijlsma acknowledges NPS-Euronet for his post-doctoral fellowship
Lisbon, 28th September 2016

19. Identification and Assessment of New Psychoactive Substances: A European Network
Identification and assessment of new psychoactive substances: a European network - NPS-Euronet (Home/2014/JDRU/AG/DRUG/7086)
Félix Hernández
Research Institute for Pesticides and Water (IUPA)
Laboratory of Pesticide Residue Analysis (LARP)
Universitat Jaume I, Castellón, Spain
Lisbon, 28th September 2016