Detection of the pharmaceutical agent

1
Detection of the pharmaceutical agent Glaucine
as a recreational drug
Analytical Unit St Georges - University of
London, London, UK1 Guys and St Thomas Poisons
Unit, London, UK2
Jennifer BUTTON1, Paul I DARGAN2, Leonard
HAWKINS2, Hanna OVASKA2, Satnam LIDDER2, John
RAMSEY1, David M WOOD2, David W HOLT1
/
INTRODUCTION
EXPERIMENTAL
RESULTS
Materials Glaucine (Glauvent) was obtained in
tablet form (40mg) from Sopharma, Bulgaria.
Flurazepam, obtained from Sigma-Aldrich (Poole,
Dorset, England), was used as an internal
standard (IS). Sodium hydroxide (40 solution)
and phosphoric acid were also obtained from BDH.
HPLC grade methyl-tert-butyl-ether (MTBE) was
purchased from Rathburn Chemicals Limited
(Walkerburn, Scotland).
Glaucine is an anti-tussive agent, which is used
therapeutically in several countries in the
European Union, such as Romania and Bulgaria, as
well as neighbouring countries, including Iceland
and Russia. Therapeutic use of glaucine has been
reported to be associated with dissociative type
symptoms 1. Animal studies suggest that
glaucine has central dopaminergic effects 1,2.
Glaucine is a marketed pharmaceutical this is
the first reported case of confirmed toxicity
associated with its recreational use. Case
Study A 23 year old female presented to the
emergency department (ED) following ingestion of
two tablets of Head Candy, a 1-benzylpiperazine
(BZP) free herbal high, marketed as a legal
alternative to conventional recreational drugs of
abuse such as MDMA (ecstasy) and amfetamine,
which had been purchased from a local head
shop. She developed nausea and vomiting within
30 minutes of ingestion, followed by a period of
dissociative type symptoms, feeling detached
and in another world. On arrival in the ED she
was agitated, vomiting, tachycardic (100 bpm) and
tachyphnoeic, but with a normal blood pressure
(135/82 mmHg) and temperature (36.8?C). Her
pupils were dilated (6mm), although the remainder
of her neurological examination was normal.
Abdominal examination was unremarkable. She was
treated with intravenous fluids and anti-emetics
(prochlorperazine 12.5mg IM and cyclizine 50mg
IV) and admitted overnight for observation. She
was discharged the following day once her
symptoms had resolved.
Glaucine and flurazepam were quantified
monitoring their most abundant ion m/z 354 and
86 and their retention times were 19.09 and 17.75
minutes, respectively.
Figure 3. NIST Library Ion Spectra for Glaucine.
(a)
(b)
Figure 2. 40mg Glaucine (Glauvent) Sopharma,
Bulgaria. TICTAC Communications Ltd
Sample preparation The sample and calibrators
(250µL) were prepared using liquid-liquid
extraction. They were adjusted to an alkaline pH
with 1M sodium hydroxide (250µL), after the
addition of 100µL of the IS flurazepam (1mg/L).
The solution was extracted with 4mL MTBE.
Following centrifugation the organic layer was
transferred to 0.1M phosphoric acid (250µL) and
mixed. After phase separation by centrifugation
the organic layer was removed to waste. 1M sodium
hydroxide (100µL) and MTBE (200µL) were added to
the remaining subnatent. The samples were then
vortex mixed (30 seconds) and centrifuged. An
aliquot of the supernatant was injected onto the
GC-MS system. Gas chromatography-mass
spectrometry GC-MS analysis was performed using a
Shimadzu GC-MS-QP2010 with a Shimadzu AOC-20i
autosampler. An HP-5 MS (30m x 0.25mm, 0.5µm
5-Phenyl)-methylpolysiloxane) analytical column
(Agilent, Palo Alto, California), was employed
for separation. Helium was used as the carrier
gas at a flow rate of 1mL per minute. The
injection volume was 1.0µL and injections were
made in splitless mode. The injector was
maintained at 225C and the detector at 200C.
The initial column temperature was set at 80C
and held for 4 minutes. It was then ramped by
25C per minute up to 290C and held for 9.6
minutes, giving a total run time of 22 minutes.
Positive Electron Impact Ionisation (EI) mode was
used and data were collected using single ion
monitoring (SIM).
Figure 4. (a) Chromatogram (b) Calibration curve,
quadratic curve fit applied
CONCLUSION
Glaucine is a marketed pharmaceutical this is
the first reported case of confirmed toxicity
associated with its recreational use. In
addition this case highlights that it is being
included by manufacturers in legally sold
BZP-free alternatives to established
recreational drugs. In our view legislative
authorities should be aware of the potential
diversion of this pharmaceutical into the
recreational drug scene, and adapt the licensing
of this product.
Figure 1. London Underground Mind Music Red Eye
Frog, BZP Free, Herbal High Party Pill
containing Glaucine only. TICTAC Communications
Ltd
Samples collected at the time of admission, were
sent to the Forensic Toxicology Service, St
Georges, University of London for analysis.
Glaucine was detected in both the urine and serum
(approximately 0.7mg/L) samples. There are no
previously published therapeutic or toxic
concentrations for comparison. Extended
toxicological screening did not detect any other
drugs or ethanol, except for cyclizine that was
administered in the ED. Due to the
unavailability of a pure reference standard,
Glaucine was obtained in tablet formation for
confirmation and quantification. The measured
concentration should therefore only be considered
an approximation.
References
1 Rovinskia VI. (2006) Acute glaucine syndrome
in the physician's practice the clinical picture
and potential danger. Klin Med (Mosk)
8468-70 2 Asencio M Hurtado-Guzmán C, López
JJ, et al. (2005) Structure-affinity
relationships of halogenated predicentrine and
glaucine derivatives at D1 and D2 dopaminergic
receptors halogenation and D1 receptor
selectivity. Bioorg Med Chem 133699-3704