Feasibility of FTIR Chemical Imaging

for Forensic Analysis of Suspected

Illicit Materials on Blotter Papers:

LSD vs. 25C-NBOMe

Presented by: Sabra Botch-Jones, MS, MA, D-ABFT-FT

Boston University School of Medicine, Biomedical Forensic Sciences

Presented to: Separation Science/Spectroscopy Solutions:

Advances in Forensics & Toxicology eSeminar

Date: October 24, 2017

Overview• Background: Pharmacological

Effects/Pattern of Use

• Historical to Novel Approaches:

GC-MS and Ambient Ionization

High Resolution Time-of-Flight

Mass Spectrometry (TOFMS)

• Feasibility of FTIR Chemical

Imaging

Background September 27, 2016, US DEA made

three of these compounds (25I-, 25C-,

and 25B-NBOMe) Schedule I on the

Controlled Substances Act (CSA)*

Similar to Shulgin’s 2C-I, 2C-C, and 2C-

B, differ by the addition of a 2-

methoxybenzyl group on the nitrogen

(NBOMe)

On-line presence 2010/2011 onwards

“Legal” alternative to LSD or not?

5-HT2A

*https://www.deadiversion.usdoj.gov/fed_regs/rules/2016/fr0927_2.htm

• September of 2013, 22 year-old/white-male found during a welfare

check

• Apparent trauma to forehead; lacerations behind the right ear;

contusion on the left side of the decedents chest and punctures on

the side of the right and left sides of the neck were observed.

• The individual’s own hair was found in the mouth and teeth.

• Toxicology-Negative

• Cause of Death: Exsanguination Due to Sharp Force Injury of Ear

• Manner of Death: Undetermined

Homicide or Not?

25C-NBOMe25I-NBOMe 25B-NBOMe

Most Common Online

Most Published

25H-NBOMe

25D-NBOMe

25E-NBOMe

Mescaline-NB2OMe.HCl

(Mescaline-NBOMe.HCl)

25T7-NB2OMe.HCl

(2C-T-7-NBOMe.HCl)

25T4-NB2OMe.HCl

(2C-T-4-NBOMe.HCl)

25T2-NB2OMe.HCl

(2C-T-2-NBOMe.HCl)

25P-NB2OMe.HCl

(2C-P-NBOMe.HCl)

25N-NB2OMe.HCl

(2C-N-NBOMe.HCl)

Sublingual/Oral/Buccal Nasal Insufflation

IV, IM, Vaginal, Anal, Smoked

Routes of Administration

5-HT2A

25I-NBOMe 2C-I

10-16

Receptor

Binding Affinity

What does that mean?

Potent serotonin (5-HT2A) agonists

Clinical Toxicology/ED Data

Forensic Data:

Toxicology/Drug Chemistry

Post Mortem/DUID

Self Reported Data

Animal Studies

Pharmacological Effects

Pattern Of Use

Clinical Studies

NBOMe User Avg. Age: 21

Class Hallucinogens Avg.

Age: 26

Lawn, et. al. “The NBOMe hallucinogenic drug series: Pattern of use, characteristics of users and self-reported effects

in a large international sample”. Journal of Psychopharmacology 2014, Vol. 28(8) 780-788

Self Reported Data: The Global Drug Survey

NBOMe Hallucinogenic Drug Series

NBOMe Users by Country

USA

UK

Euro-Zone

Australia

Canada

3%

56%

21%

10%

10%

25I-NBOMe Deaths

World Health Organization,

Expert Committee on Drug Dependence 36th Meeting, June 2014

*Some Unconfirmed

‘Pleasurable High’ ‘Negative Effects While

High’

‘Strength of Effect’ ‘Risk of Harm Following

Use’

‘Comedown After Use’

‘Urge to Use More’

Subjective Effects: Self Reported Data

Global Drug Survey

Classic Hallucinogens Dissociative Drug (i.e. Ketamine)

Cost $1.37 (USD, 2013)

“Best value for money compared to other

hallucinogenic drugs”

National Forensic Laboratory Information System (NFLIS) 2011-2015

*Drug Enforcement Administration NFLIS Special Report: 2C-Phenethylamines, Piperazines, and Tryptamines Reported in NFLIS, 2011–

2015 https://www.nflis.deadiversion.usdoj.gov/DesktopModules/ReportDownloads/Reports/NFLIS-SR-2CPEA-PiperazineTrypt.pdf

Current Analytical Procedures

GC/MS and LC/MS

• Soak blotter paper in

0.5mL of methanol

for 6hrs

– For LC: must also

dilute with 0.1% (v/v)

formic acid in water

GC/MS with derivatization, FTIR, and NMR

• Soak blotter paper in 1.0mL methanol in an ultrasonic bath for 1hr

• Extract evaporated until dried under a stream of air.

– For NMR, extract dissolved in deuterated chloroform

– Derivatization steps:

1. dissolve extract in 100µl of derivitizing agent (TFAA:chloroform, 1:1, v:v)

2. vortex

3. incubate at 70°C for 40mins

4. cool to room temp

5. evaporate to dryness with air stream at 37°C

6. reconstitute with 80µl of ethyl acetate

https://ac.els-cdn.com/S0379073816303383/1-s2.0-S0379073816303383-main.pdf?_tid=95a4c9ea-a7ad-11e7-

be0c-00000aacb361&acdnat=1506975144_0355639491df3317a673d3f5bcef9e6f

A Faster Route for Screening

• DSA-TOFMS

– Direct Sample Analysis-Time of

Flight Mass Spectrometer

• PerkinElmer Axion2TOFMS

Liquid measurement- 10µL spot

NBOMe’s Investigated

Most Common Form

• 25I (25C)

Less Common Forms

• 25D

• 25E

• 25H

Newly Investigated

Formulations

• 25T2

• 25T4

• 25T7

• 25P

• Mescaline

Materials

Results-Experiment #3

Blotter paper containing suspected LSD seized in Maine.

Spectrum Spotlight 400

Spotlight: Components for IR

Detector in LN2

Dewar

Torroidal Mirror

Cassegrains

IR beam from

spectrometer

Aperture position

Z-fold optics

Motorized

sample

stage

Blotter paper containing suspected

LSD seized in Maine.

NBOMe & LSD Treated Blotter Paper

ATR Imaging

Measurement Conditions

– Scan range: 4,000-748 cm-1

– Spectral resolution: 8 cm-1

– Scans per pixel: 4

– Field of view: 500 x 500 um

– Spatial resolution: 1.56 um

– Measures spectra: 102,400

Untreated Paper PCA

Cellulose

Polyethylene/carbonate mixture

Untreated Paper PCA

Hydrocarbon/Fluorocarbon mixture

Hydrocarbon/Fluorocarbon/Carbonate

Mixture

Overlay Comparison

LSD reference

25C-NBOMe Reference

Untreated Paper

Spectral matching of reference spectra to each measured spectrum in image

Untreated paper correlation image to LSD Untreated paper correlation image to 25C-NBOMe

LSD Treated Paper PCA

LSD/ paper components mixture

LSD reference spectrum

LSD Treated Paper Correlation Image

Comparison of Correlation Images

Untreated Paper LSD Treaded Paper

NBOMe Treated Paper PCA

Hydro / Fluorocarbon

Mixture Cellulose

Hydrocarbon/Carbonate

Mixture

NBOMe treated paper

correlation image

Some evidence of NBOMe present

Not a good agreement with “show structure” image

Correlation Image –Highest Correlation ~ 0.78

NBOMe reference

Conclusions• For the screening of NBOMe designer drug variables on blotter paper, DSA-

TOFMS and FTIR was successful at reducing analysis time from minutes to

seconds for qualitative analyte identification for selected analytes of interest.

• Application to a forensic sample was successful, further demonstrating its utility

in the forensic laboratory with these types of compounds.

• There are indications that ATR imaging may be used to identify the presences of

LSD and NBOMe on a paper matrix

• High spatial resolution is needed to isolate the chemical entity from the paper

constituents

• The use of high spatial resolution results in a small field of view or area

measured which may limit the ability to examine the whole matrix

• A challenge with low dose drug on blotter paper is if drug is not well dispersed

within paper matrix, technique may produce a negative result do to limitation of

area measured

• No detection levels have been determined for this technique and would need to

be conducted with further research.

Acknowledgements• Frank Kero-PerkinElmer

• Jamie Foss-PerkinElmer

• Thomas Byron-PerkinElmer

• Michelle Laussen-PerkinElmer

• Douglas Townsend-PerkinElmer

• Jill A. Koepke-PerkinElmer

• Maine Health and Environmental Testing Laboratory

• David Barajas-Boston University School of Medicine

• Boston University School of Medicine, Biomedical Forensic

Sciences

References1. Lawn, et. al. “The NBOMe hallucinogenic drug series: Pattern of use, characteristics of users and self-reported effects in a large

international sample”. Journal of Psychopharmacology 2014, Vol. 28(8) 780-788

2. Expert Committee on Drug Dependence. “25I‐NBOMe Critical Review Report Agenda Item 4.19” World Health Organization,

Expert Committee on Drug Dependence 36th Meeting June 2014 http://www.who.int/medicines/areas/quality_safety/4_19_review.pdf

3. Walterscheid, et. al. “Pathological Findings in 2 Cases of Fatal 25I-NBOMe Toxicity” Am J Forensic Med Pathol. Vol. 35 . No. 1 ,

March 2014

4. Johnson, RD, et. al. “An Evaluation of 25B-, 25C-, 25D-, 25H-, 25I-, and 25T2-NBOMe via LC/MS/MS: Method Validation and Analyte

Stability”. Robert D. Johnson; Sabra R. Botch-Jones, Tiffany Flowers, Connie Lewis. Journal of Analytical Toxicology 2014 (In press

5. Poklis, J. et. al. “Postmortem detection of 25I-NBOMe [2-(4-iodo-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl)methyl]ethanamine] in

fluids and tissues determined by high performace liquid chromatography with tandem mass spectrometry from a traumatic death”

Forensic Science International 234 (2014) e14-e20

6. Halberstadt, A., et. al. “Effects of the hallucinogen 2,5-dimethoxy-4-iodophenethylamine (2C-1) and superpotent N-benzyl derivatives

on the head twich response”. Neuropharmacology 77 (2014) 200-207

7. Bersani, F.S. et. al. “ 25C-NBOMe: Preliminary Data on Pharmacology Psychoactive Effects, and Toxicity of a New Potent and

Dangerous Hallucinogenic Drug”. BioMed Research International. Vol 2014, Article ID 734749

8. Zuba, et. al. 25C-NBOMe – New potent hallucinogenic substance identified on the drug market”. Forensic Science International 227

(2013) 7-1

9. Poklis. J., et. al. “Determination of 4-bromo-2, 5-dimethoxy-N-[2-methoxyphenyl methyl]-benzeneethanamine (25B-NBOMe) in serum

and urine by high performance Liquid chromatography with tandem mass spectrometry in a case of severe intoxication”. Drug Test Anal.

Jul 2014; 6(0): 764-769

10. Stellpflug, et. al. “2-(4-Iodo-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl)methyl]Ethanamine (25I-NBOME): Clinical Case with

Unique Confirmatory Testing”. J. Med. Toxicol. (2014) 10:45-50

11. Forrester, M. “NBOMe Designer Drug Exposure Reported to Texas Poison Centers”. Journal of Addictive Diseases. 12 Aug 2014 (In

Press)

Questions?

Sabra Botch-Jones M.S., M.A., D-ABFT-FT

Department of Anatomy & Neurobiology

Boston University School of Medicine

Biomedical Forensic Sciences

72 E. Concord St., L805C

Boston, MA 02118

617-638-1969 (Office)

sabraj@bu.edu

www.bumc.bu.edu/gms/biomedforensic/