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 1

FSF Emerging Forensic Scientist Award Oral Presentation 2

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. 3

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 4

Smart Drugs • Pharmacokinetics – Oral administration – Rapid absorption – Half life: 1-6 hours • Pharmacodynamics?? http://www.smarternootropics.com/2014/06/the-best- nootropic-dont-believe-the-hype/ – Increases adrenergic, dopaminergic, and histaminergic activity in the brain – Inhibit the release of γ -aminobutyric acid (GABA) 5

Abuse Potential • Enhances – Focus – Memory – Vigilance – Attention http://adhdrevamp.com/tag/smart-drugs/ • Increases drive and motivation • Does not cause restlessness and involuntary motor movements 6

Adverse Effects • Aggression • Depression • Insomnia • Anxiety • Headaches • Fainting http://www.standard.co.uk/lifestyle/esmagazine/a-quarter-of-my-friends-use-themthe- oxbridge-students-addicted-to-brainboosting-smart-drugs-8901879.html 7

Culture and Availability • Media Influence – Lucy (2014) – Limitless (2011) • Online Markets – nootropics.co.uk – nootropicssupply.com – smartpowder.com 8

Culture and Availability 9

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 10

Drugs of Interest Piracetam Pramiracetam Aniracetam Ciproxifan Noopept 11

Drugs of Interest Modafinil Adrafinil 12

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 13

Gas Chromatography/Mass Spectrometry (GC/MS) • Agilent 6890/5973 GC/MS system 14

Liquid Chromatography/Mass Spectrometry (LC/MS) • Agilent 1100 Series LC/MSD 15

LC Parameters Parameter Condition Column C18; 3.5 µm; 4.6 mm x 100 mm Solvent A: 10 mM Ammonium Acetate, pH 4 Mobile Phase 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 16

LC/MS Detection Retention Molecular Ionization Fragmentor Compound Time (min.) Ion Mode 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 17

10,000 ng/mL Piracetam Chromatogram of Neat Mix Pramiracetam Ciproxifan Aniracetam Noopept Adrafinil Modafinil Methadone D3 18

Concentration/Response Evaluation 0.250 R 2 Compound Ciproxifan 0.9973 0.200 Adrafinil 0.9909 Modafinil 0.9833 Area Ratio Aniracetam 0.9851 0.150 Ciproxifan Piracetam 0.9908 Adrafinil Modafinil 0.100 Aniracetam Piracetam 0.050 0.000 0 5 10 15 20 Concentration (ng/µL) 19

Concentration/Response Evaluation 1.600 R 2 Compound Noopept 0.9993 1.400 Pramiracetam 0.9977 1.200 1.000 Area Ratio 0.800 Noopept Pramiracetam 0.600 0.400 0.200 0.000 0 5 10 15 20 Concentration (ng/µL) 20

Supported Liquid Extraction (SLE) 1. Prepare columns 4 – Diatomaceous earth 3 1 – 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 21

5000 ng/mL Matrix Chromatogram of Mix Extracted from Piracetam Pramiracetam Human Blood Ciproxifan Aniracetam Noopept Adrafinil Modafinil 22 Methadone D3

5000 ng/mL Matrix Chromatogram of Mix Extracted from Piracetam Pramiracetam Human Urine Ciproxifan Aniracetam Noopept Adrafinil Modafinil Methadone D3 23

Limit of Detection – Blood • Racetams Typical Daily Plasma Literature Limit of Nootropic Dose Concentrations 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 *prodrug for piracetam 24

Limit of Detection – Blood • Modafinil class Typical Daily Serum Literature Limit of Nootropic Dose Concentrations Detection (LOD) Adrafinil* 900 mg 5 mg/L 0.2 mg/L 13 – 18 mg/L Modafinil 200 mg 0.7 mg/L Urine: 3.6 – 9.9 mg/L *prodrug for modafinil 25

Limit of Detection – Blood • Others Typical Daily Literature Limit of Nootropic Dose Concentrations Detection (LOD) Noopept - - 0.004 mg/L Ciproxifan - - 0.007 mg/L 26

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 28

Future Work • Validation of LC/MS method • Quantitate smart drugs in biological samples and pill/powder products

Acknowledgements • Arcadia University • Center for Forensic Science Research & Education • Forensic Sciences – Mandi Mohr, MSFS Mentoring Institute – Melissa Friscia, MSFS – Rini Gupta – Warren Korn, BS – Iryna Kurochka – Francis Diamond, BS – Dymere Taylor • Forensic Sciences – Fredric Rieders Family Renaissance Foundation Foundation 30

FSF Emerging Forensic Scientist Award Oral Presentation 31

QUESTIONS? Mollie Mares molliemares@gmail.com 32