Technical Note
Identification
of a New Amphetamine Type Stimulant:
3,4-Methylenedioxy-N-(2 hydroxyethyl)amphetamine (MDHOET)
Carola Koper*
Netherlands Forensic Institute
P. O. Box 24044
2490 AA The Hague, The Netherlands
[email: c.koper -at- nfi.minjus.nl]
Elisa Ali Tolppa
National Bureau of Investigation
P.O. Box 285
FIN-01301 Vantaa, Finland
Joseph S. Bozenko Jr.
U.S. Department of Justice
Drug Enforcement Administration
Special Testing and Research Laboratory
22624 Dulles Summit Court
Dulles, Virginia 20166, USA
Valérie
Dufey
Laboratoire Police Scientifique de Lyon
31 Avenue Franklin Roosevelt
69134 Ecully, France
Michael Puetz
Bundeskriminalamt
Thaerstrasse 11
65193 Wiesbaden, Germany
Céline
Weyermann
Institute de Police Scientifique
University of Lausanne, Batiment de Chimie
CH 1015 Lausanne Dorigny, Switzerland
Frantisek Zrcek
Police of Czech Republic
Institute of Criminalistics Prague
P.O. Box 62/KUP/Strojnicka 27
17089 Prague, Czech Republic
ABSTRACT: 3,4-Methylenedioxy-N-(2-hydroxyethyl)amphetamine
(MDHOET), an MDA derivative, was identified in Ecstasy-type tablets
seized in France, and subsequently in exhibits seized in Austria, The
Netherlands, Switzerland, and the United Kingdom. This unusual amphetamine
type stimulant (ATS) was submitted as an unknown to seven European
laboratories participating in a sponsored ATS profiling program. Six
of the seven laboratories successfully identified MDHOET upon initial
analysis. Analytical data from gas chromatography, infrared spectroscopy,
mass spectrometry, and nuclear magnetic resonance spectroscopy are
presented.
KEYWORDS: 3,4-Methylenedioxy-N-(2
hydroxyethyl)amphetamine, MDHOET, MDMA, Ecstasy, ATS, CHAMP, Forensic
Chemistry
Introduction
Photo 1 |
In
December 2004, one thousand white Ecstasy-type tablets with the “Euro” logo
were seized in Saint Etienne, France (see Photo 1).
These tablets were subsequently determined to actually contain a combination
of 3,4-methylenedioxymethamphetamine
(MDMA) and 3,4- methylenedioxy-N-(2-hydroxyethyl)amphetamine (MDHOET;
see Figure 1) [1,2].
Subsequent to this initial submission, six additional seizures containing
MDHOET were made in Europe (see Table 1), including
powders in the Netherlands (three separate submissions), white tablets
with a “LOVE” logo (no photo) in Austria and Switzerland,
and fragments of tablets in the United Kingdom. MDMA was present as
a co-ingredient in five of the seven cases, but when present was always
at a lower percentage versus MDHOET.
Figure
1. 3,4-Methylenedioxy-N-(2 hydroxyethyl)amphetamine
(MDHOET)
MDHOET
can be synthesized similarly to MDMA; for example, by reductive amination
of 3,4-methylenedioxyphenyl-2-propanone (piperonylmethylketone
or PMK) with ethanolamine, using a reducing agent (e.g., sodium cyanoborohydride)
or via catalytic hydrogenation (e.g., H2 over Platinum)
[1,2].
Not much is known about the physiological effects of this drug; it
is reported
to have very limited activity, presumably due to its relatively high
polarity [2]. It is unknown whether the drug was intentionally
synthesized as a non-controlled “designer drug,” or instead
was an erroneous synthesis of 3,4-methylenedioxyethylamphetamine (MDEA);
that
is, by mistakenly using ethanolamine instead of ethylamine.
As
MDHOET had not, to our knowledge, been previously encountered in
Ecstasy-type
tablets or powders, it represented an ideal test compound
for submission to the seven laboratories currently participating in
a project entitled: “Collaborative Harmonisation of Methods for
Profiling of Amphetamine Type Stimulants” (CHAMP). Exhibits of
the “Euro” tablets seized in France were used as the test
samples.
Experimental
Gas Chromatography - Mass Spectrometry (GC/MS)
An Agilent 6890 GC coupled to a 5973 Mass Selective Detector system
(MSD) was used. The column that was used was a HP Ultra-1 (length:
12 m, inner diameter: 0.22 mm, film thickness: 0.3 µm). Helium
was used as the carrier gas (1 mL/minute, split ratio 50:1). The
GC oven was programmed from 110°C (1 minute hold) to 275 °C
at a rate of 40 °C/minute. The carrier gas velocity was set at
40 cm/second (1.0 mL/minute, constant flow rate) and the inlet temperature
was
set at 275 °C. The injected volume was 1 µL. The scan range
was m/z 35 to 450. A solvent delay of 0.8 minute was applied.
The temperature of the MS transfer line was 300 °C.
Liquid Chromatography Mass Spectrometry (LC MS/MS)
An Agilent 1100 Series HPLC system with autosampler and an Agilent
1100 series LC/MSD Trap ion trap MS was used, with Agilent LC/MSD
Trap software version 5.2 (Bremen, Germany). The column was a Phenomenex
Luna C18 (3 mm x 150 mm, 3 mm). The eluent A was 0.01 M ammonium
acetate with 0.1 % formic acid and eluent B was acetonitrile with
5 % 0.01 M ammonium acetate and 0.1 % formic acid, in gradient run.
The gradient used was 20-100 % eluant B over 0-10 minutes and 100
% eluant B from 10-25 minutes. The flow rate was 0.3 mL/minute, and
the column temperature was 30 °C. The injection volume was 10 µL.
The
electrospray ionisation ESI technique was used, in the positive ion
mode. Operating parameters of the ESI ion source were as follows:
Drying gas temperature was 350 °C, drying gas flow 9.0 L/minute,
nebuliser gas pressure 40 psi, end plate voltage -3500 V, and end plate
offset
-500 V. Ion trap parameters were as follows: Accumulation time was
43 ms and averages 5, rolling averaging off, and ion charge control
on.
The fragmentation amplitude was increased from 30 % to 200 % from the
set value of 1.0 V. AutoMS(n) mode was used. The scan range was m/z 50
- 500.
Fourier Transform Infrared Spectroscopy (FT IR)
A Thermo Nicolet Nexus 670 was used. Scans were recorded from 4000
cm -1 to 400 cm -1; an average of 32 scans was taken. Spectra were
obtained using an attenuated total reflectance (ATR) attachment (data
not corrected).
Nuclear Magnetic Resonance Spectroscopy
Analyses (see Figures 6 and 7) were performed on a Varian Mercury 400
MHz NMR using a Varian Nalorac 5 mm indirect detection, pulse field
gradient (PFG), variable temperature probe, with PulseTuneTM. The
sample was prepared at 26.8 mg/mL in deuterium oxide (D2O) containing
TSP (3-(trimethylsilyl)propionic 2,2,3,3-d4 acid, sodium salt) as
the 0 ppm reference, and maleic acid (5 mg/mL) as the internal standard
for quantitation (maleic acid exhibits a singlet at 6.4 ppm). The
proton spectrum of the standard used to determine the purity of the
synthesized reference compound was obtained with 8 scans using a
45 second delay, 90° pulse, 5 second acquisition time, and oversampling
of 4. Confirmation of the structure of the synthesized reference
compound was performed using proton, carbon, COSY, HSQC, and HMBC
NMR spectra, and Advanced Chemistry Developments Structure Elucidator
software program (ACD/Labs, Toronto, Canada).
Synthesis of Reference Material
MDHOET was synthesized by reductive amination of 3,4-methylenedioxyphenyl-2-propanone
(PMK, 4.52 grams) with ethanolamine hydrochloride (25 grams) in
isopropanol (IPA) and sodium cyanoborohydride (NaCNBH3,
1.1 grams) as the reducing agent [1]. PMK was added
to ethanolamine hydrochloride in IPA and vigorously stirred. NaCNBH3 was
added and the mixture was stirred, adjusting the pH (as needed) with
HCl in
IPA at room temperature. After 3 days, 300 mL of water was added,
and the mixture was made strongly acidic with 37 % HCl. The resulting
solution was extracted 3 times with 100 mL CH2Cl2,
which was discarded. The remaining aqueous phase was then made basic
with 25 % NaOH, then
extracted 3 times with 100 mL CH2Cl2. The resulting
organic extracts were combined and dried over anhydrous Na2SO4.
The solvent was removed
under vacuum, yielding a clear, slightly viscous oil (2.82 grams).
This oil was dissolved in IPA containing HCl and diluted with diethyl
ether, precipitating 3.85 grams (68 %) MDHOET HCl as an off-white
powder.
Results and Discussion
Laboratory Results
Subexhibits of the “Euro” tablets seized in Saint Etienne,
France were distributed to the participating laboratories as “blind” samples
(that is, with no indication that they were a test, and no instructions
on recommended methods for analysis). This resulted in the use of a
variety of techniques, of which GC/FID, GC/MS, FT-IR were the most
frequently employed (see Table 2). The composition
of the tablets (approximately 2 % MDMA (as base), caffeine, and approximately
7 % MDHOET (as base)
was an unusual complicating factor. Six of the seven partners properly
identified MDHOET; the lone exception missed its presence during initial
screening due to its co-elution with caffeine during the GC/FID analysis
used in their laboratory. To confirm the identification, one laboratory
synthesized a reference standard, a second laboratory used NMR spectroscopy
(1H-, 13C- , COSY and HETCOR), and a third laboratory
did both. In addition to MDMA and MDHOET, caffeine, sorbitol and cellulose
were identified
by some of the laboratories (using GC, 1H-NMR, and/or FT-IR).
Mass Spectrometry
All laboratories taking part in the round robin used mass spectrometry
for the elucidation of the structure of the unknown analyte. Apart
from GC/MS, both liquid chromatography - mass spectrometry (LC/MS)
and capillary electrophoresis - mass spectrometry (CE/MS) were used.
When LC/MS, CE/MS or ion trap GC/MS was used, the M+H ion was present
at m/z 224 (Figure 2). However, if quadrupole GC/MS was used, derivatization
(for example, silylation or acetylation) was necessary to determine
the molecular ion.
Upon
GC/MS analysis of a non derivatized sample, both MDMA (Retention
index (RI) 1515) and MDHOET (RI 1865) were detected [3].
The mass spectrum of MDHOET is displayed in Figure
3. No molecular
ion could be identified.
Besides the base peak at m/z 88 (CH3CH=+NHCH2CH2OH
), other characteristic fragment ions are visible at m/z 44, m/z 70
(-18 (H20) from m/z 88),
m/z 135 (-88 (CH3CH=+NHCH2CH2OH))
and m/z 163 (-60 (H2NCH2CH2OH)). The two fragments at m/z 70
and m/z 88 both suggest the presence of
an OH functionality (that is, loss of H20).
Acetylation
of MDHOET gave two derivatives, in agreement with the presence of
two active hydrogens (NH and OH). The major compound was
the di-acetylated derivative (Figure 4, molecular
ion at m/z 307),
while the minor compound was the mono-acetylated compound (molecular
ion at m/z 281; spectrum not shown). Comparison of the mass
spectrum of MDHOET with that of its amphetamine analogue, N-(2-hydroxyethyl)amphetamine
as described by Cry et al. and Carpenter et al. [4,5],
further corroborates this interpretation.
Fourier Transform Infrared Spectrometry
The FTIR-ATR spectrum of the synthesized MDHOET HCl is shown in Figure
5. The principal wavebands are at 1031 and 1249 cm-1, with additional
characteristic bands at 3369, 2949 and 1578 cm -1.
NMR Spectroscopy
The 1H and 13C NMR spectra of the synthesized
MDHOET in D2O
are shown in Figures 6 and 7. The two active hydrogens
are not visible due
to deuterium exchange with the solvent (when dissolved in CDCl3,
both resonances are visible as broad singlets between 9.2-9.3 ppm).
The 1H-NMR spectrum of the original sample dissolved
in D2O is shown in Figure 8. In addition to MDHOET,
MDMA, sorbitol, and caffeine
are observed. The assignment of both the 1H and 13C
resonances of MDHOET are given in Table 3.
Conclusions
Although
MDHOET is quite unlikely to ever become a significant drug of abuse,
future
encounters are probable, both in Europe and elsewhere.
The analytical data presented in this article should enable facile
analyses of this unusual “designer drug,” in accordance
with SWGDRUG protocols [6].
Acknowledgements
The
Sixth Framework Program of the European Commission is gratefully
acknowledged for
funding this project. The authors would like to thank
the following persons for their contributions: Laura Aalberg (Vantaa,
Finland), Fabrice Besacier (Ecully, France), Jiri Bolehovsky (Prague,
Czech Republic), Rainer Dahlenburg (Wiesbaden, Germany), Susanne Dieckmann
(Wiesbaden, Germany), Laurence Dujourdy (Ecully, France), Pierre Esseiva
(Lausanne-Dorigny, Switzerland), Céline Delaporte (Lausanne-Dorigny,
Switzerland), Henk Huizer (The Hague, The Netherlands), Libuse Kawulokova
(Prague, Czech Republic), Eric Lock (The Hague, The Netherlands), Anneke
Poortman (The Hague, The Netherlands), Milan Prazak (Prague, Czech
Republic), Erkki Sippola (Vantaa, Finland), and Pavel Tomicek (Prague,
Czech Republic) [See address header for respective addresses.]
References
1.
Braun U, Shulgin AT, Braun G. Research on the central activity and
analgesia of N-substituted analogs of the amphetamine derivative
3,4-methylenedioxyphenylisopropylamine. Arzneimittelforschung 1980;30(5):825-30.
2. Shulgin A, Shulgin A. PIHKAL. Transform Press, Berkeley, California,
1991, p. 731.
3.
Kovats E. Gas chromatographische charakterisierung organischer Verbindungen
Teil 1: Retentionsindices aliphatischer halogenide, alkohole,
aldehyde, und ketone. Helvetica Chimica Acta 1958;41:1915-1932.
4.
Cry TD, Dawson BA, By AW, Neville GA, Shurvell HF. Structural elucidation
of unusual police exhibits. II. Identification and spectral characterization
of N-(2-hydroxyethyl)amphetamine hydrochloride. Journal of Forensic
Sciences 1996;41(4):608-611.
5.
Carpenter J, Hugerl J, Weaver K. The identification of N-(2-hydroxyethyl)amphetamine.
Journal of the Canadian Society of Forensic Science 1993;26(4):143-146.
6. Scientific Working Group for the Analysis of Seized Drugs (SWGDRUG)
Recommendations. www.swgdrug.org/approved.htm
Table
1. Seizures
of MDHOET in Europe During the Time Frame December 2004-March 2006.
| Country |
Date |
Logo |
Wt., Diam., Width |
Contents |
Amount |
| France |
12/04 |
Euro |
282 mg, 9.1 mm, 3.6 mm |
MDHOET, MDMA, caffeine |
1000 tablets |
| Netherlands |
02/05 |
- |
- |
MDHOET, MDMA |
10 g powder |
| 06/05 |
- |
- |
MDHOET, MDMA |
20 g powder |
| 03/06 |
- |
- |
MDHOET, MDMA |
1.8 g powder |
| Austria |
05/05 |
Love |
190 mg, 6.7 mm, 4.1 mm |
MDHOET |
50 tablets |
| Switzerland |
05/05 |
Love |
278 mg, 7.1 mm, 4.5 mm |
MDHOET, MDMA |
Not Reported |
| U.K. |
04/05 |
Prob.Unmarked |
- |
MDHOET |
Tablet Fragments |
Table
2. Overview
of the Techniques Used to Identify MDHOET.
LAB |
GC/FID |
HPLC |
GC/MS |
GC/MS |
LC MS/MS |
DLC |
CE MS/MS |
FTIR |
NMR |
Synthesis |
Deriv. |
1 |
X |
|
Quad |
X |
X |
|
|
X |
|
|
2 |
X |
|
Quad |
X |
|
X |
|
X |
|
X |
3 |
|
|
Quad |
X |
|
|
|
|
|
|
4 |
X |
X |
Quad |
X |
|
|
|
|
|
|
5 |
X |
|
Quad/Ion Trap |
|
|
|
|
X |
|
|
6 |
|
|
Ion Trap |
|
|
|
X |
X |
X |
|
7 |
X |
X |
Quad |
X |
|
|
|
X |
X |
X |
Table
3. Assignment of 1H and 13C Resonances of MDHOET
| Position |
Proton (ppm, Peak Muliplicity, Coupling Constant) |
Carbon (ppm) |
| CH3 |
1.28 (d, J = 6.6 Hz) |
18.0 |
| CH3-CH |
3.6 (ddq, J = 5.8, 8.7, 6.6 (x3)) |
58.4 |
| CH3-CH-CH2 |
3.07 (dd, J = 13.8, 5.8 Hz), 2.80 (dd, J = 13.8, 8.7 Hz) |
41.2 |
| N-CH2-CH2-OH |
3.28 (td, J = 13.3, 5.3 Hz), 3.21 (td, J = 13.1, 5.2 Hz) |
49.1 |
| N-CH2-CH2-OH |
3.85 (t, J = 5.2 Hz) |
59.7 |
| O-CH2-O |
5.96 (s) |
104.1 |
| phenyl #1 |
n/a |
132.5 |
| phenyl #2 |
6.85 (d, J = 1.47 Hz) |
112.6 |
| phenyl #3 |
n/a |
150.4 |
| phenyl #4 |
n/a |
149.2 |
| phenyl #5 |
6.88 (d, J = 7.92 Hz) |
111.6 |
| phenyl #6 |
6.80 (dd, J = 7.92, 1.47 Hz) |
125.7 |
| NH, OH |
Exchanged with Solvent |
N/A |
Figure
2. LC-MS/MS Mass Spectrum of MDHOET HCl.
Figure 3. Mass Spectrum of MDHOET HCl.
Figure
4. Mass Spectrum of the Double Acetylated Derivative
of MDHOET HCl.
Figure
5. Uncorrected FTIR-ATR Spectrum of Synthesized
Reference MDHOET HCl.
Figure
6. Proton NMR Spectrum of Synthesized Reference MDHOET HCl in D2O.
Figure
7. Carbon NMR Spectrum of Synthesized Reference
MDHOET HCl in D2O.
Figure
8. Proton NMR Spectrum of one of the French “Euro” Tablet
in D2O
(Containing MDHOET HCl, MDMA HCl, Caffeine, and Sorbitol).
