Development of an Analytical Method for Nootropic Smart Drugs in - - PowerPoint PPT Presentation

Development of an Analytical Method for Nootropic ‘Smart’ Drugs in Biological Fluids

Mollie Mares, BS*; Karen Scott, PhD; Donna Papsun, MS; Barry Logan, PhD

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FSF Emerging Forensic Scientist Award Oral Presentation

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Disclaimer

• This presentation describes applications

developed on Agilent 6890/5973 GC/MS system and Agilent 1100 Series LC/MSD.

• Agilent Technologies provided no financial

support for this project.

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Smart Drugs

• Nootropics
• Stimulants but alleged to boost brain function

and cognition

• Uses:

– Alzheimer’s disease – Parkinson’s disease – ADHD – “Academic doping”

http://uanews.org/story/good-vibrations-mediating- mood-through-brain-ultrasound

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Smart Drugs

• Pharmacokinetics

– Oral administration – Rapid absorption – Half life: 1-6 hours

• Pharmacodynamics??

– Increases adrenergic, dopaminergic, and histaminergic activity in the brain – Inhibit the release of γ-aminobutyric acid (GABA)

http://www.smarternootropics.com/2014/06/the-best- nootropic-dont-believe-the-hype/

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Abuse Potential

• Enhances

– Focus – Memory – Vigilance – Attention

• Increases drive and motivation
• Does not cause restlessness and involuntary

motor movements

http://adhdrevamp.com/tag/smart-drugs/

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Adverse Effects

• Aggression
• Depression
• Insomnia
• Anxiety
• Headaches
• Fainting

http://www.standard.co.uk/lifestyle/esmagazine/a-quarter-of-my-friends-use-themthe-

• xbridge-students-addicted-to-brainboosting-smart-drugs-8901879.html

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Culture and Availability

• Media Influence

– Lucy (2014) – Limitless (2011)

• Online Markets

– nootropics.co.uk – nootropicssupply.com – smartpowder.com

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Culture and Availability

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Significance

• Smart drugs are sold online and in illicit supply

chains

• Most have not been approved or scheduled in

the US

• Concern about the adverse effects in relation

to public health and safety

• Not routinely tested in forensic casework

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Drugs of Interest

Piracetam Pramiracetam Aniracetam Noopept Ciproxifan

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Drugs of Interest

Modafinil Adrafinil

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Research Objectives

• Development of a single analytical method for

the detection and quantification of smart drugs in blood, urine, and pill/powder products

• Validation of the assay for the analysis of

unknown samples

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Gas Chromatography/Mass Spectrometry (GC/MS)

• Agilent 6890/5973 GC/MS system

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Liquid Chromatography/Mass Spectrometry (LC/MS)

• Agilent 1100 Series LC/MSD

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LC Parameters

Parameter Condition

Column C18; 3.5 µm; 4.6 mm x 100 mm Mobile Phase Solvent A: 10 mM Ammonium Acetate, pH 4 Solvent B: 50:50 Acetonitrile:Isopropanol Gradient 90% A, 10% B to 10% A, 90% B Run time 15 minutes Flow Rate 0.6 mL/min Injection Volume 10 µL

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LC/MS Detection

Compound Retention Time (min.) Molecular Ion Ionization Mode Fragmentor Voltage

Piracetam 2.86 143 Positive 70 Pramiracetam 6.48 270 Positive 70 Aniracetam 10.92 220 Positive 100 Noopept 11.53 319 Positive 100 Ciproxifan 10.67 271 Positive 130 Modafinil 12.09 272 Negative 80 Adrafinil 11.96 288 Negative 90 Methadone-D3 12.80 313 Positive 80

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Chromatogram of Neat Mix

Piracetam Pramiracetam Aniracetam Modafinil Adrafinil Ciproxifan Noopept Methadone D3 10,000 ng/mL

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Concentration/Response Evaluation

0.000 0.050 0.100 0.150 0.200 0.250 5 10 15 20

Area Ratio Concentration (ng/µL)

Ciproxifan Adrafinil Modafinil Aniracetam Piracetam

Compound R2

Ciproxifan 0.9973 Adrafinil 0.9909 Modafinil 0.9833 Aniracetam 0.9851 Piracetam 0.9908

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0.000 0.200 0.400 0.600 0.800 1.000 1.200 1.400 1.600 5 10 15 20

Area Ratio Concentration (ng/µL)

Noopept Pramiracetam

Concentration/Response Evaluation

Compound R2

Noopept 0.9993 Pramiracetam 0.9977

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Supported Liquid Extraction (SLE)

• 1. Prepare columns

– Diatomaceous earth – Glass wool

• 2. Prepare sample (200 uL)

– 1:1 matrix:pH9 phosphate buffer

• 3. Absorption of sample
• 4. Solvent elution

– 75:25 DCM:IPA

• 5. Dry down and reconstitute in mobile phase

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1 3 4

Chromatogram of Mix Extracted from Human Blood

Piracetam Pramiracetam Aniracetam Modafinil Adrafinil Ciproxifan Noopept Methadone D3 5000 ng/mL Matrix

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Chromatogram of Mix Extracted from Human Urine

Piracetam Pramiracetam Aniracetam Modafinil Adrafinil Ciproxifan Noopept Methadone D3 5000 ng/mL Matrix

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Limit of Detection – Blood

Nootropic Typical Daily Dose Plasma Literature Concentrations Limit of Detection (LOD)

Pramiracetam* 1000 – 1500 mg

• 0.005 mg/L

Piracetam 1500 – 6500 mg 27 – 264 mg/L 0.7 mg/L Aniracetam 400 – 1200 mg 0.0087 – 0.014 mg/L 0.04 mg/L

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• Racetams

*prodrug for piracetam

Limit of Detection – Blood

Nootropic Typical Daily Dose Serum Literature Concentrations Limit of Detection (LOD)

Adrafinil* 900 mg 5 mg/L 0.2 mg/L Modafinil 200 mg 13 – 18 mg/L Urine: 3.6 – 9.9 mg/L 0.7 mg/L

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• Modafinil class

*prodrug for modafinil

Limit of Detection – Blood

Nootropic Typical Daily Dose Literature Concentrations Limit of Detection (LOD)

Noopept

• 0.004 mg/L

Ciproxifan

• 0.007 mg/L

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• Others

Powders and Capsules

Nootropic Type Recommended Dosage Confirmed

Piracetam Powder 1500 – 6500 mg Yes Pramiracetam Capsule 1200 mg Yes Aniracetam Powder 1500 mg Yes Adrafinil Powder 900 mg Yes

Conclusions

• Developed a single analytical method for the

identification of smart drugs in blood and urine using LC/MS and SLE

– Percent recoveries around 50-70%

• Detected smart drugs in pill and powder

products

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Future Work

• Validation of LC/MS method
• Quantitate smart drugs in biological samples

and pill/powder products

Acknowledgements

• Arcadia University
• Forensic Sciences

Mentoring Institute

– Rini Gupta – Iryna Kurochka – Dymere Taylor – Fredric Rieders Family Renaissance Foundation

• Center for Forensic Science

Research & Education

– Mandi Mohr, MSFS – Melissa Friscia, MSFS – Warren Korn, BS – Francis Diamond, BS

• Forensic Sciences

Foundation

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FSF Emerging Forensic Scientist Award Oral Presentation

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Mollie Mares molliemares@gmail.com

QUESTIONS?