SCREENING OF NEW PSYCHOACTIVE SUBSTANCES

IN URINE SAMPLES

BY HIGH RESOLUTION MASS SPECTROMETRY

Noelia Salgueiro-González, Sara Castiglioni, Emma Gracia-Lor, Nikolaos I. Rousis, Lubertus Bijlsma, Alberto Celma, Félix Hernández, Ettore Zuccato

INDEX

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

ANALYSIS OF URINE SAMPLES

CONCLUSIONS

Acknowledgements

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

INTRODUCTION

ANALYSIS OF URINE SAMPLES

• Alcohol• Drugs of abuse• Hallucinogens• Antidepressant drugs• Nicotine• Caffeine

Regulation and/oroutlawing

Psychoactive Substances

Natural or synthetic substances capable to affectmental proccesses when they are administrated

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

ANALYSIS OF URINE SAMPLES

CONCLUSIONS

Acknowledgements

INTRODUCTION

New Psychoactive Substances

Derived from controlleddrugs molecules

Similar psychoactiveproperties of drugs of abuse

High number NPS availableNot regulated

High potential toxicity for human health!

Use of NPS increased in lastyears

2C-B (Phenethylamine)

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

INTRODUCTION

More tan 450 NPS150 NPS between 2014-2015

ANALYSIS OF URINE SAMPLES

Groups of NPS

Synthetic cannabinoids

Synthetic cathinones

Phenethylamines

Synthetic opioids

Tryptamines

Piperidines

Aminorex derivatives

Natural NPS

Benzodiazepines

Ketamine analogues

Synthethic Cathinones

Synthetic cannabinoids

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

INTRODUCTION

ANALYSIS OF URINE SAMPLES

Investigations based on NPS

Epidemiologicalsurveys

Analysis of urinesamples

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

INTRODUCTION

ANALYSIS OF URINE SAMPLES

Investigations based on NPS

Quantitativeanalysis

Qualitativeanalysis

Number of NPS analyzed limited

Not all standards are available

Some metabolites unkwon

Screening of high number of compounds/metabolites

Less analysis time

High Resolution mass spectrometry

Promising tool for NPS identification

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

OBJECTIVES

Evaluation of the consumption of NPS indifferent European cities

Screening of the 190 selected NPS in pooledurine samples collected during music festivals

Development of instrumental methodology forthe screening of NPS by liquid chromatographyhigh resolution mass spectrometry (LC-HR-MS)

ANALYSIS OF URINE SAMPLES

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

ANALYTICAL METHODOLOGY

5 mL of pooled urinesample

Solid Phase extraction (SPE)HLB cartridges. Elution with 3 mL

MeOH

Determination LC-HR-MSMobile phase: Water (0.1%

formic acid) + ACNColumna: X Bridge® C18,

2.1x100 mm, 3.5 µm

ANALYSIS OF URINE SAMPLES

Steps of the analytical method

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

ANALYTICAL METHODOLOGY

Determination LC-HR-MS

Step 1: ScreeningAdquisition mode: Full MS

Step 2: Suspected screeningAdquisition mode : Full MS/ddMSn

Step 3: Identification of suspectedAdquisition mode : Full MS/DIAOrbitrap QExactive®

Confirmation of suspected:Information found in literatureAnalytical Standards available

ANALYSIS OF URINE SAMPLES

Software Xcalibur®

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

ANALYSIS OF URINE SAMPLES

ANALYSIS OF URINE SAMPLES

Pooled Urine samples from European festivals

Three pooled urine samples

European festivals

Collected from portable toilets

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

ANALYTICAL METHODOLOGY

ANALYSIS OF URINE SAMPLES

Synthetic cannabinoids

Synthetic cathinones

Phenethylamines

Synthetic opioids

Tryptamines

Piperidines

Aminorex derivatives

Natural NPS

Benzodiazepines

Ketamine analogues

61

53

39

9

7

7

3

4

4

3

Screening of NPS

190 NPS

Frequency of detection reported by the European Early Warning System

Software Trace Finder®

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

ANALYSIS OF URINE SAMPLES

ANALYSIS OF URINE SAMPLES

Retention time

Fragments

Exact mass

Pooled urine sample. Identificationand confirmation of 4-Cl-alpha-PPP

238.0994 (Exact mass)

4-Cl-alpha-PPP

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

ANALYSIS OF URINE SAMPLES

ANALYSIS OF URINE SAMPLES

Standard 4-Cl-alpha-PPP

Exact massFragments

Retention time

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

ANALYSIS OF URINE SAMPLES

ANALYSIS OF URINE SAMPLES

Fragments m/zStandard

(Ms)

Fragments m/zsample(Mm)

Error mass(Mm-Ms)/Ms

Confirmation

238.0994 238.0989 -2.0999

167.0259 167.0259 0

139.0311 139.0308 -2.1578

98.0970 - -

Confirmation of the presence of 4-Cl-alpha-PPP in one urine sample!!

• Retention time agreement. Error>2%

• Exact mass. Error mass < 5 ppm

• 2 fragments identified (Error mass <5 ppm)

Complex matrix

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

CONCLUSIONS

The investigation of selected NPS in urine samples wascarried out to evaluate the consumption of thesesubstances in European festivals

An analytical method based on high resolution massspectrometry was proposed, being an interesting tool forthe screening of NPS in urine samples

Among the 190 substances investigated, only the syntheticcathinone 4-Cl-α-PPP was identified and confirmed in theanalyzed samples by acquiring the corresponding analyticalstandard.

As fas as we know, this is the first time 4-Cl-alpha-PPP wasidentified in urine sample from music festivals. However,further research is needed to know its concentration.

ANALYSIS OF URINE SAMPLES

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

Acknowledgements

Postdoctoral fellowshipN.Salgueiro-González

Postdoctoral fellowshipHolder center

Research Project:HOME/2014/JDRUG/AG/DRUG/7085

ANALYSIS OF URINE SAMPLES

Conference grantN.Salgueiro-González

Other Research projects:GRC2013-047

CTM2013-48194-C3-2-R

INTRODUCTION

OBJECTIVES

ANALYTICAL METHODOLOGY

CONCLUSIONS

Acknowledgements

Acknowledgements

THANK YOU FOR

YOUR ATTENTION!!!

ANALYSIS OF URINE SAMPLES

ANY QUESTIONS?

Noelia Salgueiro GonzálezPostdoctoral researcher

E-mail: noelia.salgueiro@marionegri.it

http://www.npseuronet.eu/

For more information