1) FTIR chemical imaging spectroscopy and DSA-TOFMS were used to analyze fortified blotter paper samples containing NBOMe designer drugs and LSD without sample preparation. 2) FTIR was able to spatially separate LSD from paper components using a library match. Preliminary results also separated NBOMe from paper but with lower correlation. 3) High spatial resolution is needed to isolate drugs from paper but limits the analysis area. Additional case samples are needed to validate the method for this matrix.

1. The Screening and Confirmation of NBOMe Designer Drugs on Blotter Paper by Ambient Ionization Mass Spectrometry and FTIR Chemical Imaging Spectroscopy
Amanda Moore1 , Sabra Botch-Jones1, David Barajas1, Tom Byron2, Ryan Smith2, Jamie Foss2,3, Jason Weisenseel2, Kevin Tyvoll2, Frank Kero2
1. Boston University School of Medicine, Biomedical Forensic Sciences, 72 E. Concord St., Boston, MA 02118
2. PerkinElmer Environmental Health , 2651 Warrenville Road, Ste. 100 , Downers Grove, IL 60515
3. Maine Health and Environmental Testing Laboratory, 221 State St., Augusta, ME 04333
Novel Aspects
No prep analysis of fortified blotter paper samples for illicit designer drugs of
abuse using FTIR chemical imaging for orthogonal analysis versus ambient
ionization mass spectrometry.
MAFS 2016 *For complete method details, contact: frank.kero@perkinelmer.com
LSD/ paper components mixture
LSD reference spectrum
NBOMe reference
Figure 12: Spatial determination of illicit substances by FTIR library matching
NBOMes
Figure 2. Structures of relevant designer drugs in this study
• NBOMe drugs (Figure 2) are also known as new
psychoactive substances (NPS).
• N-(methoxybenzyl)-phenethylamine
compounds are typically consumed on blotter
paper.
• The user experience has been described as a
more potent, but similar high when compared
to LSD.
• These substances are emerging as an issue of
public health as an increase in cause of death
case studies have found a link to NBOMes.
• Reference spectra is provided in Figure 10
LSD
Introduction
Previous work from this group: N-(methoxybenzyl)-phenethylamine
compounds (also known as “NBOMe’s”) are an emerging threat to public health
with fatality and severe injury related investigations increasing throughout the
country. The NBOMe compounds are obtained as “legal” highs through
unregulated internet purchases and are typically administered via blotter paper.
The user experience has been described as a more potent, but a similar high
when compared to lysergic acid diethylamide (LSD). Users often believe they
are ingesting LSD when in fact it was one or a combination of the NBOMe
compounds. Previous reports from this group have demonstrated the utility of
non-targeted high resolution mass spectrometry platforms for this application
paired with both ambient ionization and electrospray ionization sources since
authentic case work samples are often obtained as mixtures of true unknowns
(reported at NEAFS 2015). This previous work employed a minimal sample
preparation via a 5 min liquid extraction of the target compound(s) from the
blotter paper. This work was completed in collaboration with the State of Maine
Department of Health and Human Services (Augusta, Maine), the University of
Central Florida (Orlando, Florida ) and Boston University School of Medicine
Biomedical Forensic Sciences Program (Boston, Massachusetts).
The next step: In an extension of this work, chemical imaging using Fourier
Transform Infrared Spectroscopy (FTIR) was evaluated to provide an orthogonal
workflow solution for confirmation and to leverage the advantage of Direct
Sample Analysis-Time-of-Flight Mass Spectrometry (DSA-TOFMS, PerkinElmer,
Waltham, MA, USA) for screening (e.g.to reduce the speed of analysis, reduced
cost in consumables and waste etc.). FTIR chemical imaging method
development variables considered included variation in sampling depth and
scan modes to generate a spatial chemical map of the sample. Method:
Certified reference material of 25C-NBOME (Cerilliant, Round Rock, TX, USA)
was analyzed as “cast” films by allowing a liquid solution in methanol to
evaporate before analysis. In addition, 25C-NBOMe was spiked onto blotter
paper. Further, certified reference material of LSD (Cerilliant) was also analyzed
for comparison. A macro Geranium (Ge) Attenuated Total Reflectance (ATR)
sampling accessory was used. The scan range was 4,000-748 cm-1, spectral
resolution was 8 cm-1, scans per pixel was 4, field of view was 500 x 500 um,
special resolution 1.56 µm, measures spectra of 102,4000, and the total
number of scans were 64. By utilizing the Ge ATR, the diffraction limits were 2.5
um at 1,000 cm-1.
Utilizing the spectral library and chemical imaging, LSD was identified and able
to be separated from the blotter paper components which were identified as
polyethylene/carbonate, cellulose, hydrocarbon/fluorocarbon/carbonate
mixtures. The correlation for the NBOMe was determined to be ~0.78 and the
same components were identified from the blotter paper. There are indications
that this technique 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, however it results in a small field of view or
area measured limiting the ability to examine the whole matrix. If drug is not
well dispersed within paper matrix, technique may produce a negative result do
to limitation of area measured. It is anticipated that this workflow would be of
broad-based interest to the forensic community on the topic of emerging drugs
of abuse, designer and synthetic drugs. More data on case samples is needed to
confirm conclusions on the utility of FTIR imaging for this matrix.
Figure 1: Chemical Structure of LSD
• Lysergic acid diethylamide = LSD (Figure 1)
• Well reported hallucinogenic substance
• Culture of abuse emerged in the 1950’s
• Clear or white odorless material
• Sublingual dosage on blotter paper
• Reference spectra is provided in Figure 10.
An overview of the DSA source and instrument housing is
provided in Figures 5 and 5.
Method details
• Probe Temp: 300°C
• Nebulizer N2 Gas flow: 80psi
• Aux Gas: 4 L/min
• Dry Gas Heater 25°C
• Corona current: 6 µAmp
• Scan rate: 10 scans / second
• Cap entrance: -800 V
• Endplate: -200 V
• Capillary Exit Voltage: 120 V
• Note: DSA uses APCI tune mix for lock mass ions for daily
calibration of the TOFMS
Figure 4. DSA source schematic
Figure 5. DSA-TOFMS (left) DSA (top right) DSA ion source with mesh target screen (bottom right)
Blotter paper samples
Figure 4. Samples as received
• Blotter paper samples were fortified at Boston University (Figure 3) to support
method development.
• This concentration was selected to be comparable with current dosages relevant for
consumption.2
• Solution standards obtained from Cayman Chemical (Ann Arbor, Michigan) were
included in the shipment for method development (Figure 4)
Figure 3. a) blotter paper b) fortified at Boston University
Untreated paper NBOMe treaded paper LSD treaded paper
Figure 11 Chemical imaging of fortified blotter paper with hallucinogenic drugs of abuse
Analysis of an Authentic Forensic Case Sample: Suspected LSD
[C18H22ClNO3 +H]+
[C8H9O]+
[C7H7]+
Figure 8. Fragmentation pathway to m/z 121 and m/z 91 ions
Figure 7. (Top) Singe dosage unit analyzed under initial
instrument conditions with a capillary exit setting of 100V (10
μL sample volume). Mass Accuracy = 1.2 ppm. (Bottom) Single
dosage unit analyzed under adjusted instrument conditions
with a capillary exit setting of 120V showing CID fragmentation
(10 μL sample volume). Mass accuracy = 0.60 ppm.
Figure 6. Suspected LSD blotter
paper seized in Maine.
Under the hood
• Does not require N2 purge
• MIR/NIR
• 1.56µm spatial resolution
• Mode switching
• Hot and cold stage options
• Software automated atmospheric
compensation
Figure 9: FTIR Chemical Imaging instrumentation
PerkinElmer
Spotlight 400
A)
B)
Real case sample: A perforated blotter paper was seized in Bangor, ME
and submitted to the Maine Health and Environmental Testing Laboratory
as suspected LSD for analysis by DSA-TOFMS.
• Sample prep – 5 min liquid extraction with DCM
• Sample volume for analysis - 10 µL
• The analysis time per replicate was 15 sec
DSA-TOFMS identified the sample as 25C-NBOMe. Results were confirmed by
GC-MS….. This method has a relatively long analysis time by comparison.
Q: Can no prep FTIR imaging do better? A: Possibly (more data needed to confirm)
Figure 10: FTIR reference spectra for the
Selected analytes
Please note: This DSA-TOFMS case study has recently been published in Forensic Science International 267 (2016) 89–95
Stand alone FTIR reference spectra
LSD
25C-NBOMe