Piperazine-like compounds: a new group of designer

drugs-of-abuse on the European market

Douwe de Boer
a,b,*
, Ingrid J. Bosman
b
, Elod Hidvegi
c
, Carmo Manzoni
a
,
Andras A. Benko
c
, Lourenco J.A.L. dos Reys
a
, Robert A.A. Maes
b
a
Instituto Nacional do Desporto, Laboratorio de Analises de Dopagem e Bioqu mica,
Av. Prof. Egas Moniz (Estadio Universitario), 1600-190 Lisbon, Portugal
b
Human Toxicology, Faculty of Pharmacy, Utrecht University, Utrecht, The Netherlands
c
National Institute for Forensic Toxicology, Budapest, Hungary
Abstract
1-Aryl-piperazine compounds are, depending on their substituents, selective for certain serotonin receptors and together
with their easy availability and their so-called legal status, this group of psychoactive compounds are potential designer drugs-
of-abuse. Internet in that respect is an important source of information and distribution facilities. Because this development
may have consequences for the interpretation of future clinical and forensic toxicological case studies, some analytical aspects
of 1-benzyl-piperazine (BZP), 1-[4-methoxyphenyl]-piperazine (pMeOPP) and 1-[3-triuoromethylphenyl]-piperazine
(TFMPP) were studied. BZP was not detected by the AxSYM
1
FPIA technology designed to determine amphetamine-like
compounds, but had showed some cross reactivity with EMIT
1
d.a.u.
1
. The cross reactivities at 300 and 12,000 ng/ml (RS)-
amphetamine equivalents were 0.4 and 1.3%, respectively. Although BZP was not identied directly by the REMEDi
TM
HS
Drug Proling System, it can be detected by this HPLC/UV scanning system. Using GC/NPD without derivatisation, BZP,
pMeOPP and TFMPP can be analysed for and applying GC/MS without or with acetylation or triuoroacetylation, these
compounds can be identied unambiguously. The usefulness of GC/NPD and GC/MS in this respect was demonstrated by the
quantitative and qualitative analysis of the content of a capsule with the synthetic stimulant A2, which proved to contain
86.4 mg of BZP. # 2001 Elsevier Science Ireland Ltd. All rights reserved.
Keywords: 1-Benzyl-piperazine; 1-[4-Methoxyphenyl]-piperazine; 1-[3-Triuoromethylphenyl]-piperazine
1. Introduction
Several examples of compounds that contain a piperazine
moiety in their molecule bind to serotonin receptors. Appar-
ently the structures of piperazine-like compounds have good
interaction possibilities with these receptors, which can be
understood if orientations of for example 1-aryl-piperazines
are compared with that of serotonin [1]. Although as a group
the piperazine-like compounds cannot be considered as
selective compounds for serotonin receptors, they may be
made more site selective with the appropriate substituents
[2]. Together with their easy availability and their so-called
legal status, this group of psychoactive compounds are
therefore potential designer drugs-of-abuse.
An illustration of this phenomenon are the 1-aryl-piper-
azines. An example is 1-benzyl-piperazine (BZP, Fig. 1A),
which roughly mimics the psychoactive effects of (S)-
amphetamine, although at a higher dosage [3,4]. Other
examples of this group of psychoactive compounds are 1-
[3-chlorophenyl]-piperazine (mCPP, Fig. 1B) [5], 1-[4-
methoxyphenyl]-piperazine (pMeOPP, Fig. 1C) [6] and 1-
[3-triuoromethylphenyl]-piperazine (TFMPP, Fig. 1D) [7].
All of the examples mentioned are promoted as drugs-of-
abuse on the Internet. BZP is since January 2000 commer-
cially available on a European Internet website as a so-called
synthetic stimulant. It is sold in capsules under the name of
A2 in a dosage of 125 mg of BZP dihydrochloride. The other
examples mCPP, pMeOPP and TFMPP are only sold as a
free base or as a salt in semi-bulk quantities and not yet in
pharmaceutical formulations. Although only partly sup-
ported by literature, TFMPP has the reputation to mimic
Forensic Science International 121 (2001) 4756
*
Corresponding author.
0379-0738/01/$ see front matter # 2001 Elsevier Science Ireland Ltd. All rights reserved.
PII: S0 3 7 9 - 0 7 3 8 ( 0 1 ) 0 0 4 5 2 - 2
the psychoactive effects of 3,4-methylenedioxymethamphe-
tamine (MDMA) [7].
As this development may have consequences for the
interpretation of future clinical and forensic toxicological
case studies, the respective toxicologists should be aware of
the possibility to encounter cases with the abuse of BZP,
mCPP, pMeOPP and TFMPP. Therefore, in this study certain
analytical strategies to determine their abuse will be eval-
uated and the corresponding analytical data will be pre-
sented. Attention will be mainly focussed on BZP as well as
on the capsules with the synthetic stimulant A2. Also, some
analytical aspects of pMeOPP and TFMPP will be presented.
mCPP was not investigated in this context.
2. Materials studied, methods, techniques
2.1. Materials and stock solutions
The piperazine-like compounds analysed were from
Lancaster (Lancaster Synthesis Ltd., Strasbourg, France);
1-benzyl-piperazine dihydrochloride 98%, 1-[2-methylphe-
nyl]-piperazine (oMePP) dihydrochloride 98%, 1-[4-
methylphenyl]-piperazine (pMePP) dihydrochloride 97%,
1-[2-methoxyphenyl]-piperazine (oMeOPP) hydrochloride
98%, 1-[3-methoxyphenyl]-piperazine (mMeOPP) dihy-
drochloride 97%, 1-[4-methoxyphenyl]-piperazine dihy-
drochloride, 1-[3-triuoromethylphenyl]-piperazine 98%.
Stock solutions were prepared at concentrations of 1 mg/
ml in demineralised water, except for BZP and TFMPP,
which were dissolved in methanol. All other reagents and
chemical used were of analytical grade. The capsules with
the synthetic stimulant A2 were ordered through Internet at
the website of Smart Drugstore International, a so-called
supplier of lifestyle products on Internet.
2.2. Isolation of compound of interest from capsules
The content of the capsule as far as possible was dissolved
in 100 ml of demineralised water (stock solution). Two
hundred and forty microliter of a diluted stock solution
(diluted 1:4) were added to 5 ml of demineralised water.
The aqueous phase was made basic with 0.5 ml of 5 MKOH.
After adding 3 g of Na
2
SO
4
, a liquid/liquid extraction was
performed with 2 ml of tert-butyl methyl ether in order to
isolate the compound of interest.
2.3. Immunoassays
Two different immunoassays were evaluated. All assays
were performed according to the manufacturers' recommen-
dations. The enzyme-multiple immunoassay technique
(EMIT
1
) was performed with the d.a.u.
1
Amphetamines
using the ETS
1
Plus System analyser from Syva
1
(Dade
Behring Inc., Deereld, IL, USA). The uorescence polar-
isation immunoassay (FPIA) was performed with the
AxSYM Amphetamine/Methamphetamine II assay using
the AxSYM
1
system robotic analyser from Abbott Diag-
nostics System (Abbott Laboratories, Abbott Park, IL,
USA).
2.4. HPLC/UV analysis
HPLC/UV analysis was performed with an automated
analytical procedure using the REMEDi
TM
HS Drug Prol-
ing System from Bio-Rad Clinical Systems Division (Bio-
Rad Laboratories Inc., Hercules, CA, USA). The procedure
was performed according to the manufacturer's recommen-
dations using a urine sample spiked at a concentration of
10 mg/ml.
2.5. GC/NPD analysis
GC/NPD analysis was performed with a HP GC 5890
series II (Hewlett Packard, Palo Alto, CA, USA) equipped
with HP-5MS column (length 14 m, inner diameter 0.25 mm
and lm thickness 0.25 mm) and a nitrogen phosphorous
detector (NPD). The temperature during an analysis run was
maintained at 1008C for 1 min, programmed up to 3008C at
158/min and maintained there for 5 min. The temperature of
the injection port was 2508C. Helium was used as a carrier
gas at a ow rate of 1 ml/min. Ephedrine was used as an
internal standard, except for the quantitative GC/NPD ana-
lysis of the capsule with the synthetic stimulant A2 for which
pMePP was used as internal standard.
2.6. Derivatisations for GC/MS analysis
Acetylation: the acetylation was performed according
to Peger et al. [8].
Triuoroacetylation: the triuoroacetylation was per-
formed by adding 50 ml of triuoroacetic anhydride and
50 ml of ethyl acetate to a residue containing the compound
of interest. The mixture was incubated for 30 min at 658C.
Excess of reagent and organic solvent was removed under a
mild stream of nitrogen and the dry residue was re-dissolved
in ethyl acetate for GC/MS analysis.
Fig. 1. The chemical structures of (A) 1-benzyl-piperazine; (B) 1-
[3-chlorophenyl]-piperazine; (C) 1-[4-methoxyphenyl]-piperazine;
(D) 1-[3-triuoromethylphenyl]-piperazine.
48 D. de Boer et al. / Forensic Science International 121 (2001) 4756
2.7. GC/MS analysis
GC/MS analysis was performed was performed with a HP
GC 5890 series II (Hewlett Packard, Palo Alto, CA, USA)
equipped with HP-5MS column (length 19 m, inner diameter
0.25 mm and lm thickness 0.25 mm) and coupled to a HP
5971Amass selective detector (MSD) (Hewlett Packard). The
temperature during an analysis run was maintained at 908C
for 1 min, programmed up to 3008C at 158/min and main-
tained there for 5 min. The temperatures of the injection port
and transfer line were 270 and 3008C, respectively. Helium
was used as a carrier gas at a ow rate of 1 ml/min. The HP
5971A MSD operates only in the Electron Ionisation mode.
Mass spectra were recorded at the standard 70 eVin the range
of m/z 40400. Ephedrine was used as an internal standard.
3. Results and discussion
3.1. Analytical strategies
The analytical strategies in clinical and forensic toxico-
logical are principally designed to start with relatively
simple and quick screening procedures, in which immu-
noassays and chromatographic methods play an important
role. If required, a screening procedure is followed by a
conrmation method, which in many cases involves mass
spectrometry. In order to evaluate a broad range of clinical
and forensic toxicological techniques in this study, some
representative methodologies were selected such as immu-
noassay, HPLC/UV, GC/NPD and GC/MS.
3.2. Immunoassay
Because BZP acts apparently on the same receptors as
(S)-amphetamine, it was evaluated in some immunological
assays [1]. Although the binding to the same receptors does
not guarantee that the compounds of interest also interact
with the same antibodies, BZP was investigated in two
amphetamine-like immunoassays. This way antibodies with
specicities for some different epitopes of amphetamine-like
compounds were evaluated, which were relevant for the
practical work in clinical and forensic toxicology. It was
observed that at a concentration level of 100,000 ng/ml
spiked in a blank urine sample, BZP was not detected by
the AxSYM
1
FPIA technology. EMIT
1
d.a.u.
1
Ampheta-
mines however did detect BZP (Fig. 2). The cross reactivities
at 300 and 12,000 ng/ml (RS)-amphetamine equivalents
were 0.4 and 1.3%, respectively.
3.3. HPLC/UV analysis
To have an impression if HPLC/UV analysis could be
suitable to detect the abuse of piperazine-like compounds the
REMEDi
TM
HS Drug Proling System (REMEDi
TM
sys-
tem) was selected for evaluation. This system does not only
cover HPLC/UVanalysis, but also includes a way of sample
preparation. The patented approach removes several
unwanted endogenous compounds, while allowing analysis
of a wide variety of toxicologically relevant drugs. The
REMEDi
TM
system is known to be very specic, although
its sensitivity is not always considered to be adequate [9,10].
BZP was not part of the standard library of the REME-
Di
TM
system and therefore it was not identied, when a
spiked urine sample was analysed by this system. However,
the report of the REMEDi
TM
system (Table 1) indicated that
for a peak observed in the chromatogram some candidates
drugs deserved at least careful consideration, namely the
cyclic derivative of dinorpropoxyphene, 4-hydroxyphency-
clidine, alphaprodine and phencyclidine itself. The criteria
used by the REMEDi
TM
system for comparison data of the
Fig. 2. EMIT
1
d.a.u.
1
Amphetamines response rate for different concentrations of 1-benzyl-piperazine in urine.
D. de Boer et al. / Forensic Science International 121 (2001) 4756 49
unknown peak with an internal dedicated database are
retention times relative to two different internal standards
(N-ethylnordiazepam and chlorpheniramine), primary and
secondary UV absorption peaks and wavelength ratios.
According to certain algorithms possible candidates are
qualied by calculating the so-called ``similarity factor''
(SF value). For BZP the relevant REMEDi
TM
system results
are summarised in Table 1. The peak of BZP eluted between
both internal standards and had an absorption maximum in
its UV spectrum at 211 nm. This maximum is of course not
characteristic. Although not identied by the system, the
REMEDi
TM
system at least detected BZP.
3.4. GC/NPD analysis
The qualitative GC/NPD analysis of BZP showed a
prominent peak with a relative retention time of 1.23 (Table
2). Also the other compounds of interest, e.g. pMeOPP and
TFMPP, gave adequate peaks with good response factors as
can be expected from molecules with two nitrogens.
Although the peak shapes under the investigated conditions
were asymmetric and showed some tailing, the GC/NPD
analysis can be considered as an adequate screening tech-
nique.
The main identication criterion in GC/NPD analysis is
the relative retention time. If more than one compound has
the same relative retention time, a conrmation procedure
should be available for unambiguous identication. In prin-
cipal, the identication will be no problem using GC/MS,
unless isomers of the compounds exist, which have the same
or a similar mass spectrum. Therefore, it would be con-
venient to know if for example certain isomers of the
compounds of interest have same or similar relative retention
times. In this context, some isomers of BZP, the methyl-
phenylpiperazines (MePPs), and of pMeOPP were investi-
gated. Indeed, the GC/NPD analysis of oMePP showed a
similar relative retention time, namely 1.28 (Table 2). The
same conclusion could be drawn in respect to pMeOPP and
its isomer mMeOPP. Thus, when applying the GC/NPD
analysis as a screening technique, a GC/MS procedure must
be available for the conrmation to distinguish different
isomers.
3.5. GC/MS analysis
The results of the non-derivatised 1-aryl-piperazines are
summarised in Table 2. The mass spectra were relatively
simple. Besides the molecular ion the spectra are dominated
by several fragments still containing the aryl ring combined
with fragmentations at the piperazine moiety. In respect to
selectivity, i.e. possibility to distinguish isomers, the GC/MS
analysis of the non-derivatised 1-aryl-piperazines proved to
be selective for BZP and its isomers studied. The mass
spectrum of BZP was signicantly different from its isomers
in terms of relative abundancies of m/z values. For pMeOPP
and its isomer mMeOPP this difference was less signicant
and only distinction could be made based on the small
difference in relative retention times. Taking into account
also the asymmetric peak shapes and the tailing, the GC/MS
analysis of the non-derivatised 1-aryl-piperazines was there-
fore not the identication procedure of rst choice.
Acetylation changed the fragmentation pattern compared
to non-derivatisation (Table 3). The acetyl group seems to
stabilise the piperazine ring, resulting in more stable ions
and in apparently more characteristic mass spectra. In terms
of MS, there was however no improvement in respect to the
possibility to distinguish the compounds of interest and their
isomers. Under the conditions studied the peak shapes in
the GC chromatograms and the resolution improved sub-
stantially. Therefore, the selectivity in terms of GC was
better. The end result was that the GC/MS analysis of the
N-acetyl derivatives of 1-aryl-piperazines was more than
satisfactory.
The N-triuoroacetyl derivatives of the investigated 1-
aryl-piperazines (Table 4) showed to be even more stable
than the N-acetyl derivatives. This was reected by the
appearance of fragments with the intact piperazine ring.
For all of the compounds studied, except BZP, the molecular
ion was the base peak in the mass spectrum. Despite several
attempts it was impossible to obtain a common N-triuor-
oacetyl derivative of mMeOPP. Under the conditions studied
the derivatisation of mMeOPP apparently resulted in two
different bis-N-triuoroacetyl derivatives instead of one
mono-N-triuoroacetyl derivative. This problem was how-
ever not investigated in detail. In general, the selectivity was
Table 1
Results presented by the REMEDi
TM
system after the analysis of 1-benzyl-piperazine
Identity Retention time (min) L-max
a
(nm) Notes Candidates SF
b
value
Peak no. 5 2.78 233 IS1 N-ethylnordiazepam Not relevant
Peak no. 6 7.74 211 S1
c
Cyclic dinorpropoxyphene 0.245
4-Hydroxyphencyclidine 0.037
Alphaprodine 0.018
Phencyclidine 0.101
Peak no. 7 9.62 229 IS2 Chlorphenramine Not relevant
a
Wavelength of absorption maximum in UV spectrum.
b
SF: similarity factor.
c
Peak has ``one or more strong candidates'' that deserves careful consideration.
50 D. de Boer et al. / Forensic Science International 121 (2001) 4756
Table 2
GC/NPD and GC/MS results of non-derivatised 1-aryl-piperazine-like compounds as well as proposed structures of characteristic ions
a
Code of
compound
b
RRT
c
NPD
RRT
d
MS
MW m/z values of characteristic ions (normalised on base peak)
M

a b1 b2 b3 c d e
Compounds of interest
BZP 1.23 1.27 176 176 (18) 134 (67) 120 (7) 119 (2) 118 (5) 91 (100) 77 (1) 56 (20)
pMeOPP 1.73 1.88 192 192 (35) 150 (100) 136 (5) 135 (13) 134 (3) 107 (1) 56 (8)
TFMPP 1.31 1.37 230 230 (22) 188 (100) 174 (3) 173 (6) 172 (10) 145 (11) 56 (9)
Isomers of BZP
oMePP 1.28 1.32 176 176 (26) 134 (100) 120 (5) 119 (5) 118 (20) 91 (10) 56 (8)
pMePP 1.47 1.56 176 176 (29) 134 (100) 120 (5) 119 (12) 118 (9) 91 (14) 56 (8)
Isomers of pMeOPP
oMeOPP 1.51 1.61 192 192 (41) 150 (100) 136 (5) 135 (20) 134 (14) 107 (2) 56 (10)
mMeOPP 1.77 1.92 192 192 (30) 150 (100) 136 (5) 135 (9) 134 (3) 107 (3) 56 (8)
a
For the fragments of BZP n 1 and R
1
R
2
R
3
H and for those of the other 1-aryl-piperazines n 0 and the substituents R
1
, R
2
and R
3
depend on the specic structure (Fig. 1).
b
BZP: 1-benzyl-piperazine; TFMPP: 1-[3-triuoromethylphenyl]-piperazine; oMePP: 1-[2-methylphenyl]-piperazine; pMeOPP: 1-[4-methoxyphenyl]-piperazine; mMeOPP: 1-[3-methoxyphenyl]-piperazine.
c
RRT NPD: relative retention time with GC/NPD analysis (relative to ephedrine; absolute retention time of this internal standard was 4.308 min 0.004 (n 7)).
d
RRT MS: relative retention time with GC/MS analysis (relative to ephedrine; absolute retention time of this internal standard was 3.43 min 0.02 (n 7)).
Table 3
GC/MS results of N-acetyl-1-aryl-piperazine-like compounds as well as proposed structures of characteristic ions
a
Code of compound
b
RRT
c
MW m/z values of characteristic ions (normalised on base peak)
M

f g a1 a2 a3 b1 b2 b3 c d e
Compounds of interest
N-acetyl-BZP 1.12 218 218 (6) 175 (6) 146 (43) 134 (27) 133 (4) 132 (22) 120 (1) 119 (1) 118 (3) 91 (100) 77 (1) 56 (8)
N-acetyl-pMeOPP 1.36 234 234 (85) 191 (15) 162 (100) 150 (8) 149 (26) 148 (2) 136 (28) 135 (28) 134 (24) 107 (3) 56 (29)
N-acetyl-TFMPP 1.13 272 272 (30) 229 (11) 200 (100) 188 (40) 187 (13) 186 (2) 174 (8) 173 (18) 172 (28) 145 (19) 56 (29)
Isomers of BZP
N-acetyl-oMePP 1.13 218 218 (25) 175 (9) 146 (100) 134 (11) 133 (27) 132 (13) 120 (10) 119 (11) 118 (44) 91 (14) 56 (18)
N-acetyl-pMePP 1.22 218 218 (49) 175 (13) 146 (100) 134 (10) 133 (27) 132 (4) 120 (19) 119 (26) 118 (25) 91 (23) 56 (24)
Isomers of pMeOPP
N-acetyl-oMeOPP 1.25 234 234 (52) 191 (10) 162 (100) 150 (9) 149 (27) 148 (4) 136 (11) 135 (12) 134 (30) 107 (3) 56 (25)
N-acetyl-mMeOPP 1.38 234 234 (49) 191 (12) 162 (100) 150 (13) 149 (28) 148 (3) 136 (17) 135 (20) 134 (12) 107 (6) 56 (24)
a
For the fragments of BZP n 1 and R
1
R
2
R
3
H and for those of the other 1-aryl-piperazines n 0 and the substituents R
1
, R
2
and R
3
depend on the specic structure (Fig. 1); for the proposed structures of ion b1, b2, b3, c, d and e see Table 2.
b
BZP: 1-benzyl-piperazine; TFMPP: 1-[3-triuoromethylphenyl]-piperazine; oMePP: 1-[2-methylphenyl]-piperazine; pMeOPP: 1-[4-methoxyphenyl]-piperazine; mMeOPP: 1-[3-methoxyphenyl]-piperazine.
c
RRT: relative retention time (relative to N,O-acetyl-ephedrine; absolute retention time of this internal standard was 6.91 min 0.01 (n 7)).
Table 4
GC/MS results of N-triuoroacetyl-1-aryl-piperazine-like compounds as well as proposed structures of characteristic ions
a
Code of compound
b
RRT
c
MW m/z values of characteristic ions (normalised on base peak)
M

[M-CH
3
]

h i f g a1 a2 a3 b1 b2 b3 c d e
Compounds of interest
N-trifluoroacetyl-BZP 1.64 272 272 (36) 195 (16) 181 (35) 175 (14) 146 (13) 134 (2) 133 (2) 132 (9) 119 (31) 118 (20) 91 (100) 77 (1) 56 (12)
N-trifluoroacetyl-oMeOPP 2.07 288 288 (100) 273 (35) 191 (22) 162 (11) 150 (3) 149 (3) 148 (2) 136 (7) 135 (22) 134 (10) 107 (2) 56 (19)
N-trifluoroacetyl-TFMPP 1.66 326 326 (100) 229 (56) 200 (85) 188 (9) 187 (25) 186 (3) 174 (11) 175 (59) 176 (55) 145 (43) 56 (53)
Isomers of BZP
N-trifluoroacetyl-oMePP 1.63 272 272 (100) 257 (1) 175 (47) 146 (46) 134 (9) 133 (10) 132 (29) 120 (12) 119 (22) 118 (61) 91 (22) 77 (5) 56 (28)
N-trifluoroacetyl-pMePP 1.80 272 272 (100) 257 (1) 175 (41) 146 (28) 134 (5) 133 (8) 132 (4) 120 (8) 119 (31) 118 (20) 91 (22) 77 (3) 56 (22)
Isomers of pMeOPP
N-trifluoroacetyl-oMeOPP 1.86 288 288 (100) 273 (5) 191 (32) 162 (25) 150 (3) 149 (4) 148 (4) 136 (5) 135 (13) 134 (17) 107 (3) 56 (22)
N-trifluoroacetyl-mMeOPP Uncharacterised
derivatives
a
For the proposed structures of ion f, g, a1, a2, a3, b1, b2, b3, c, d and e see Tables 2 and 3.
b
BZP: 1-benzyl-piperazine; TFMPP: 1-[3-triuoromethylphenyl]-piperazine; oMePP: 1-[2-methylphenyl]-piperazine; pMeOPP: 1-[4-methoxyphenyl]-piperazine; mMeOPP: 1-[3-methoxyphenyl]-piperazine.
c
RRT: relative retention time (relative to N,O-triuoroacetyl-ephedrine; absolute retention time of this internal standard was 3.77 min 0.01 (n 7)).
as good as with the N-acetyl derivatives with the remark that
the absolute retention times were shorter.
3.6. Capsule with the synthetic stimulant A2
Qualitative analysis of the contents of a capsule with the
synthetic stimulant A2 by GC/MS analysis after triuoroa-
cetylation rapidly identied BZP as the main compound
(Fig. 3). The quantitative GC/NPD analysis without deriva-
tisation indicated that a capsule contained 86.4 mg of BZP.
This amount was in agreement with the declared amount of
125 mg of BZP dihydrochloride, corresponding to 88 mg of
the free base. The capsules were sold in pairs and assuming
that BZP mimics amphetamine at a higher dosage [3,4],
probably a pair of capsule represents one dosage.
3.7. Metabolic interpretation pitfalls
Based on this study, it can be assumed that in principal
using the appropriate toxicological methodologies the pre-
sence of 1-aryl-piperazines can be screened for and identi-
ed unambiguously. The aspect of sensitivity was not
studied here, because at this moment it is not known
which concentration levels can be expected after the admin-
istration of, for example, a capsule with the synthetic
stimulant A2.
A complication of the interpretation of analytical results is
that metabolic processes of 4-substituted 1-aryl-piperazines
also mayresult inthe presence of 1-aryl-piperazines (Table 5).
In general, it can be stated that such 1-aryl-piperazines are
detectable in human urine as minor metabolites, accounting
Fig. 3. Mass spectrum of the N-triuoroacetyl derivative of the main compound of the capsule with the synthetic stimulant A2 (upper
spectrum) vs. the N-triuoroacetyl derivative of 1-benzyl-piperazine (lower spectrum).
54 D. de Boer et al. / Forensic Science International 121 (2001) 4756
for only <10% of the administered dosage of the parent
drugs [8].
Because it may be assumed that the 4-substituted 1-aryl-
piperazines are excreted partly either as the parent com-
pounds or as some characteristic metabolites [1115], their
possible presence should be checked if one of the 1-aryl-
piperazine-like metabolites has been identied. Some of the
commercially available databases already have some infor-
mation implemented, at least of the more known 4-substi-
tuted 1-aryl-piperazines (Table 5). Only if the presence of
the parent compounds or of the characteristic metabolites
of the relevant 4-substituted 1-aryl-piperazines can be
excluded, the presence of 1-aryl-piperazines might be attrib-
uted to the use of 1-aryl-piperazines. It should be mentioned
that still some information is lacking about the metabolism
and pharmacokinetics of the drugs mentioned in Table 5.
4. Conclusions
BZP is not detected by the AxSYM
1
FPIA technology
designed to determine amphetamine-like compounds, but is
detected to some extent by EMIT
1
d.a.u.
1
.
Although BZP is not identied directly by the REME-
Di
TM
HS Drug Proling System, it can be detected by this
HPLC/UV scanning system.
In respect to GC/NPD and GC/MS, BZP, pMeOPP and
TFMPP can be identied without signicant problems.
Being aware of isomers and knowing that the compounds
of interest can be distinguished, as far as studied, from their
isomers by their mass spectra and/or retention times, the
analytical pitfalls are well covered.
With the interpretation of analytical results after the
identication of 1-aryl-piperazines in general, it should be
taken into account that the metabolism of certain 4-sub-
stituted 1-aryl-piperazine-like drugs may result in the nd-
ing of their respective 1-aryl-piperazines. The presence of
other metabolites of these 4-substituted 1-aryl-piperazine-
like drugs or of the parent drug should at least be investigated
and if necessary excluded.
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McKenney, 5-HT
1
and 5-HT
2
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Table 5
Some 4-substituted 1-aryl-piperazine-like drugs and their 1-aryl-piperazine-like metabolites
General name
of drug
Pharmacological
classification
1-Aryl-piperazine-like
metabolite
a
Literature
reference
Database information
REMEDi
TM
PMW
c
Piberaline
b
Antidepressant BZP [11,12] No No
Enziprazole
b
Antidepressant mCPP [13] No No
Etoperidone Antidepressant mCPP [14] No No
Mepiprazole Tranquilizer mCPP [13] No No
Nefazodone Antidepressant mCPP [15] Yes No
Trazodone Antidepressant mCPP [16] Yes Yes
Enciprazione
b
Anxiolytic oMeOPP [13] No No
Milipertine Antipsychotic oMeOPP [13] No No
MJ-7378
b
Antipsychotic oMeOPP [13] No No
Urapidil Antihypertensive oMeOPP [13] No No
Antrafenine Analgesic TFMPP [13] No No
a
BZP: 1-benzyl-piperazine; mCPP: 1-[3-chlorophenyl]-piperazine; oMeOPP: 1-[2-methoxyphenyl]-piperazine; TFMPP: 1-[3-triuor-
omethylphenyl]-piperazine.
b
Experimental drug.
c
PegerMaurerWeber mass spectral and GC data of drugs, poisons, pesticides, pollutants and their metabolites [8].
D. de Boer et al. / Forensic Science International 121 (2001) 4756 55
Data of Drugs, Poisons, Pesticides, Pollutants and Their
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chromatography assay with ultraviolet detection for the
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5-(1-hydroxyethyl)etoperidone and 1-(3-chlorophenyl)piper-
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56 D. de Boer et al. / Forensic Science International 121 (2001) 4756