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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??
– 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/
5
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/
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
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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
Noopept Ciproxifan
11
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
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
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 P
iraceta
m
Pra
mir
aceta
m
An
iraceta
m Mo
dafi
nil
Ad
rafi
nil
Cip
roxif
an
No
op
ep
t
Meth
ad
on
e D
3
10,000 ng/mL
18
Concentration/Response Evaluation
0.000
0.050
0.100
0.150
0.200
0.250
0 5 10 15 20
Are
a R
atio
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
19
0.000
0.200
0.400
0.600
0.800
1.000
1.200
1.400
1.600
0 5 10 15 20
Are
a R
atio
Concentration (ng/µL)
Noopept
Pramiracetam
Concentration/Response Evaluation
Compound R2
Noopept 0.9993
Pramiracetam 0.9977
20
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
Pir
aceta
m
Pra
mir
aceta
m
An
iraceta
m M
od
afi
nil
Ad
rafi
nil
Cip
roxif
an
No
op
ep
t Meth
ad
on
e D
3
5000 ng/mL
Matr
ix
22
Chromatogram of Mix Extracted from Human Urine
Pir
aceta
m
Pra
mir
aceta
m
An
iraceta
m M
od
afi
nil
Ad
rafi
nil
Cip
roxif
an
No
op
ep
t
Meth
ad
on
e D
3 5000 ng/mL
Ma
trix
23
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
24
• 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
25
• 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
28
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
30
FSF Emerging Forensic Scientist Award
Oral Presentation
31
32
Mollie Mares molliemares@gmail.com
QUESTIONS?
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