Legal Highs / Novel Psychoactive SubstancesAnalytical Strategy, HS 2019 Legal Highs Structure and Action –Legal Situation –Detection and Analytics 08.10.2018 1 Legal Highs / Novel

||Analytical Strategy, HS 2019

Legal Highs

Structure and Action – Legal Situation – Detection and Analytics

08.10.2018 1

Legal Highs / Novel Psychoactive Substances

Cedric Wüthrich, Konrad Münch 08.10.2018Cedric Wüthrich, Konrad Münch 1


Legal Highs

From Pharmaceutical Research to Legal Highs

https://www.mcmp.purdue.edu/faculty/nicholsd; Nature 469 (2011), 7

David E. Nichols

▪ Professor Emeritus of Pharmacology, Purdue University

▪ Research on Neurotransmitters (e.g. Dopamine, Serotonin), their

receptors and treatment of Parkinson’s disease

▪ Development of new antipsychotic drugs

▪ Used as reference for the development of legal highs

▪ “I was stunned I had published information that ultimately led to

human death”

08.10.2018Cedric Wüthrich, Konrad Münch 2


Legal Highs

▪ “A substance with stimulant or mood-altering properties whose sale or use is not

banned by current legislation regarding the misuse of drugs.”

Legal Highs (Oxford English Dictionary)

https://www.lexico.com/en/definition/legal_high



Legal Highs

…“Bath salts”, “Pond cleaners”, “Glass cleaners”, “Plant Food”…

Available as…



Legal Highs

…“Research chemicals”, but also “Nasal Decongestant”, plants…

Available as…



Legal Highs

▪ GABA immitates

Legal Highs – Examples (1)

Open Access Emergency Medicine (2018), 10, 9–23, Clinical Toxicology (2012), 50



Legal Highs

▪ Phenylethylamine and derivatives





Legal Highs

▪ Tryptamines and derivatives





Legal Highs

▪ Ketamines

▪ Cocaine Derivatives



▪ Methylphenidate derivatives

▪ Synthetic Cannabinoids

▪ …



Legal Highs

Can be

▪ Novel psychoactive compounds, “Designer Drugs”

▪ Over-the-counter medications (e.g. nasal decongestant)

▪ Household culinary spice and plant products (e.g. nutmeg, fly agaric mushroom)

▪ Hallucinogenic, Euphoria, Dissociative Behaviour

▪ Toxidromes like tachycardia, mydriasis, urinary retention, dry mucosal

membranes, hypoactive bowel sound, hypothermia, organ complications

▪ Often similarities with neurotransmitters

▪ Small structural changes to circumvent drug legislation

Legal Highs


Open Access Emergency Medicine (2018), 10, 9–23, Clinical Toxicology (2012), 50, The American Journal of Drug and Alcohol Abuse (2012), 38, 176–180


Legal Highs

Legal Situation in Switzerland: Betäubungsmittelgesetz, BetmG

https://www.admin.ch/opc/de/classified-compilation/20101220/index.html, https://www.admin.ch/opc/de/classified-compilation/19981989/index.html, https://www.bag.admin.ch/bag/de/home/gesetze-und-

bewilligungen/gesetzgebung/gesetzgebung-mensch-gesundheit/gesetzgebung-betaeubungsmittel.html

▪ Principle: “Prevention, Therapy, Damage

Reduction, Repression”

▪ List of illegal drugs (synthetic or natural) and

controlled substances, including

▪ Auxiliary Chemicals for production (Solvents, Acids, …)

▪ Precursors, Stereoisomers, Salts, Esters, Substitutes

(presumed similar effects)


Legal Highs

▪ Drug regulation is individual in every country

▪ Fast adoption of novel substances in member state legislation

European Monitoring Centre for Drugs and Drug Addiction

▪ Research, Data Acquisition

▪ Early warning system

▪ Fast response to novel substances

Legal Situation in EU


http://www.emcdda.europa.eu, http://www.europarl.europa.eu/news/de/press-room/20171020IPR86547/drogen-schnelles-eu-weites-verbot-von-neuen-gefahrlichen-substanzen


Legal Highs

▪ “At 1:00 a.m. a 23 year old male patient is admitted to the emergency

department of the University Hospital in Basel. He was very agitated, had a

combative violent behaviour and hallucination. He has been found in his

home standing on his bed complaining of mice everywhere. He was given the

antidote for opiate intoxications without any effect. The classical drugs of

abuse screening tests in the laboratory were negative. The physicians then

suspected the intake of legal highs or research chemicals and sent a urine

and a blood sample to the laboratory.”

Case



Legal Highs

▪ Symptomatic supportive care

▪ e.g. schizophrenia, paranoia, aggression, hypothermia, …

▪ Intravenous Fluids

▪ Drug Screening:

▪ Interferences with assays for other drugs (false-positive?)

▪ Search for unknown compound?

▪ Degradation?

▪ Probably mixture of different drugs?

Treatment


The American Journal of Drug and Alcohol Abuse (2012), 38, 176–180


Legal Highs

Requirements for the Analysis in the Problem Set

• Use of the required techniques can be performed with little

training

• Little manual work, high degree of automatization

• Little to no sample treatment

• Sophisticated instrumentation

• Instruments need to produce fast qualitative results

• Data treatment and interpretation should be performed mostly

automatically

Data

OperatorInstrument

0 1



Legal Highs

Preparing Blood and Urine Samples (1)

Urine and blood mostly contain water



Legal Highs

Preparing Blood and Urine Samples (2)

Centrifuge for removing large particles

• Precipitate proteins with acetone• Solid-phase extraction• Liquid-liquid extraction

• Solid-phase extraction• Liquid-liquid extraction

S. F. Lo, V. Young, W. J. Rhead, in Clin. Apllications Mass Spectrom. (Eds.: U. Garg, C.A. Hammett-Stabler), Humana Press, Totowa, NJ, 2010, pp. 433–443.T. M. Annesley, Clin. Chem. 2003, 49, 1041–1044.S. Tiziani, A.-H. Emwas, A. Lodi, C. Ludwig, C. M. Bunce, M. R. Viant, U. L. Günther, Anal. Biochem. 2008, 377, 16–23.

Many possibilities for extracting the aqueous layer, some methods utilize buffers



Legal Highs

Gas and Liquid Chromatography

C. F. Poole, Gas Chromatography, Elsevier, 2012.S. Fanali, P. R. Haddad, C. F. Poole, M.-L. Riekkola, Liquid Chromatography, Elsevier, 2017.

Some GC-Detectors:TCD, FID, ECD, NPD, AED, IRD, MSD

Some LC-MS-Detectors:UV/Vis, Conductivity, Refractive Index, MS



Legal Highs

Mass Spectrometer / HR-MS

J. H. Gross, Mass Spectrometry, Springer International Publishing, 2017.

𝑅 =

𝑚𝑧

Δ(𝑚𝑧 )

High resolution approximately achieved at R > 5000



Legal Highs

Ion Source / Electron Ionization




Legal Highs

MSn

Selection of precursor ion, collision, analysis of the collision products




Legal Highs

Mass Analyzer / LTQ-Orbitrap


Linear Ion Trap:Can store ions over time, select ions, can accumulate ions

C-Trap:Quadrupole, injects the ions into the orbitrap

Orbitrap:Electrostatic spindle with gradient, oscillations of ions are treated with Fourier Transformation



Legal Highs

Mass Analyzer / FT-ICR-MS

Ions in the magnetic field cause a local magnetic field in the detectors, signal is Fourier transformed




Legal Highs

Data Analysis (1)

D. Pasin, A. Cawley, S. Bidny, S. Fu, Anal. Bioanal. Chem. 2017, 409, 5821–5836.



Legal Highs

Data Analysis (2)

Top-Down

Check the TIC-chromatogram

Get molecular formula

Check database

See if MS/MS-fragments match

product

Bottom-Up

Check extracted-ion chromatogram

for known ions

Put together ions belonging to the

same peak

Generate molecular formula

D. Pasin, A. Cawley, S. Bidny, S. Fu, Anal. Bioanal. Chem. 2017, 409, 5821–5836.



Legal Highs

Conclusion


▪ Legal Highs comprise a group of psychoactive substances that are not

controlled by current legislation

▪ Treatment: Supportive Symptomatic Care

▪ Legislation was adapted to NPS (Novel Psychoactive Substances)

▪ Analytics: Sophisticated automated instrumentation, Orbitrap?

▪ Data: Target screening faster, less trained operator


Legal Highs08.10.2018Cedric Wüthrich, Konrad Münch 27

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Legal Highs / Novel Psychoactive SubstancesAnalytical Strategy, HS 2019 Legal Highs Structure and Action –Legal Situation –Detection and Analytics 08.10.2018 1 Legal Highs / Novel