Toxicology Letters 197 (2010) 157–162

Contents lists available at ScienceDirect

Toxicology Letters
journal homepage: www.elsevier.com/locate/toxlet

Mini review

Spice drugs as a new trend: Mode of action, identification and legislation

I. Vardakou, C. Pistos ∗ , Ch. Spiliopoulou
Department of Forensic Medicine and Toxicology, School of Medicine, National and Kapodistrian University of Athens, Athens 115 27, Greece

a r t i c l e i n f o a b s t r a c t

Article history: The present review highlights the existing monitoring and legislation status of synthetic cannabinoids in
Received 14 April 2010 “Spice” products and alert research community about the identification and risk assessment problems of
Received in revised form 28 May 2010 these compounds. Available data were collected by various literature search engines. All valuable infor-
Accepted 1 June 2010
mation about psychoactive properties, safety profile, clinical data and detection problems for synthetic
Available online 8 June 2010
cannabinoids and their use as “herbal highs” were managed to spot and summarise. “Spice” contains syn-
thetic cannabinoids that bind to cannabinnoid-like receptors and they are stronger than natural cannabis.
Keywords:
Chronic abuse of “Spice” has linked with signs of addiction syndrome and withdrawal symptoms similar
Spice
Synthetic cannabinoids
to syndromes observed in cannabis abuse. These cannabinoids can be considered as new products to be
JWH-018 added to the list of “designer drugs”. Although it remains unclear where and how the actual production
HU-210 of the herbal mixtures takes place, it is evident that producers are purposely risk the health of consumers
CP-47,497 to skim high profits. Only recently a number of countries in Europe, as well as in US and Canada banned
the use of these substances. The difficulty in identification of related compounds leads to the necessity
for the availability of reference standards in order to aid toxicological analyses.
© 2010 Elsevier Ireland Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157
2. Cannabinoids and their receptors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158
3. Common synthetic cannabinoids found in “Spice” products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158
3.1. JWH-018 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158
3.2. CP-47,497 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159
3.3. HU-210 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159
4. Tracking the World Wide Web and Patterns of use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159
5. Case reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159
6. Identification of synthetic cannabinoids in “Spice” products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160
7. Legal status . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
8. Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
9. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
Conflict of interest statement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161

1. Introduction ing Centre for Drugs and Drug Addiction (EMCDDA) and Europol,
found that a “smoking mixture” under the brand name “Spice” was
A number of classical cannabinoids, 9 -THC analogs, have been not the herbal product that it purported to be (EMCDDA, 2008).
synthesized based on its partially reduced dibenzopyran structure Although “Spice” was declared as “incense blend which releases
(Uchiyama et al., 2009). At the end of 2008, the European Monitor- a rich aroma” and “not for human consumption”, when smoked
as “bio-drug”, has been reported by some users to have effects
similar to those of cannabis (Auwärter et al., 2009). “Spice” and
∗ Corresponding author at: Laboratory of Forensic Medicine and Toxicology,
other “herbal” products are often referred to as “legal highs” or
School of Medicine, University of Athens, 75 Mikras Asias Str., Athens 115 27, Greece.
“herbal highs”, in reference to their legal status and purported
Tel.: +30 210 7462433; fax: +30 210 7716098. natural herbal make-up (McLachlan, 2009; Lindigkeit et al., 2009;
E-mail address: cpistos@med.uoa.gr (C. Pistos). Zimmermann et al., 2009).

0378-4274/$ – see front matter © 2010 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.toxlet.2010.06.002
158 I. Vardakou et al. / Toxicology Letters 197 (2010) 157–162
A variety of “Spice” products e.g.: Spice Silver, Spice Gold and
Spice Diamond, etc. (Auwärter et al., 2009), are mixtures (blend)
reported to contain the following plant/herbal ingredients: Indian
Warrior, Lion’s Tail, Baybean, Blue Lotus, Vanilla, Honey, etc. Based
on their chemical compositions, it can be assumed that, at least two
of these ingredients, “Indian Warrior” and “Lion’s Tail”, may have
some psychoactive effect. On the packaging of the “Spice” prod-
ucts, there is a lack of information about the complete chemical
composition and little is known about the toxicology of the plant
materials purportedly contained in the “Spice” products. Thus, no
definite answers can be provided at present, with regard to the
potential health risks related to the possible psychoactive effects,
but also in general for these products (EMCDDA, 2008).
In December 2008, chemical analysis revealed that the
psychoactive effects of the herbal mixtures were due to
added synthetic cannabinoids. Synthetic cannabinoids that
have been reported in “Spice” and “Spice”-like samples in
Europe are JWH-018, JWH-073, JWH-398, JWH-250; HU-210;
CP-47,497 and its homologues; and oleamide. The above
Cannabinoids1 /Cannabinoids2 receptor (CB1 /CB2 ) agonists are
lipid-soluble, non-polar, with typically 20–26 carbon atoms, which
are fairly volatile. Although, their detailed pharmacology has not
been investigated, it’s a fact that many of them are much more
potent than 9 -THC, therefore the usual doses may be less than
1 mg (EMCDDA, 2009a,b).
There is no evidence that JWH, CP and/or HU compounds are
present in all “Spice” products or even batches of the same prod-
uct. Different amounts or combinations of these substances seem to
have been used in different “Spice” products to produce cannabis-
like effects. It is possible that substances from these or other
chemical groups with a cannabinoid agonist or other pharmaco-
logical activity could be added to any herbal mixture (Griffiths et
al., 2010).
Up to now, the mixtures are sold in headshops and via Internet
in many countries. In several Internet blogs, drug users reported
cannabis-like effects after smoking “Spice” mixture. These products
are sold without age restriction of the clients and for that enjoy
great popularity particularly among younger people (Auwärter et
al., 2009).
This review aims to summarise any available information about
psychoactive properties, safety profile, clinical data, detection
problems and legislation for synthetic cannabinoids and their use
as “herbal highs”.
2. Cannabinoids and their receptors
Cannabinoids are a structurally diverse family of compounds
with a large number of biological targets. Based on their origins,
cannabinoids, can be classified into three groups: phytocannabi-
noids, endocannabinoids, and synthetic cannabinoids (Sun and
Bennett, 2007).
Phytocannabinoids are only known to be present in significant
quantity in the cannabis plant (Grotenhermen, 2004). The smoke
from burning cannabis contains various chemicals (Goodman and
Gilman’s, 2001). The most prevalent natural cannabinoids that
have been isolated are Tetrahydrocannabinol (9 -THC), cannabid-
iol (CBD) and cannabinol (CBN) (Grotenhermen, 2004; Pertwee,
2006).
Endocannabinoids are produced within the body and serve as
intercellular “lipid messengers”. It is believed that are synthesized
rather “on-demand” than made and stored for later use.
Synthetic cannabinoid receptor agonists, are a large family of
chemically unrelated structures, which act as 9 -THC but are more
effective (Geller, 2007). Like 9 -THC, synthetic cannabinoids bind
to the same cannabinoid receptors in the brain. Although, they were
developed as potential pharmaceutical agents, it proved difficult
to separate the desired properties from unwanted psychoactive
effects (EAMCDDA, 2009).
The effects of all three groups of cannabinoids are already known
to be generated through a specific interaction with cell membranes.
In 1980s cannabinoid receptors were identified and denoted by
the abbreviation CB and numbered in the order of their discovery
by a subscript (CB1 , CB2 ). The distinction between them is based
on differences in their predicted amino acid sequence, signalling
mechanisms, and tissue distribution (Howlett et al., 2002).
Since 1960s, analogues of 9 -THC were developed and include
HU-210 (Mechoulam et al., 1988; Glass and Northup, 1999; Ottani
and Giuliani, 2001; Jiang et al., 2005; Pertwee, 2005), Nabilone,
Dronabinol which is currently under international control (WHO
ECDD, 2006) and many others. HU-210 is reported to have 100
times the potency of 9 -THC.
Ten years later, Pfizer developed the cyclohexylphenol (CP)
series. Examples include CP-59,540, CP-47,497 and their n-alkyl
homologues (Ottani and Giuliani, 2001; Pertwee, 2005; Compton
et al., 1992, 1993).
In 1994, J.W. Huffman et al. created a large series of naph-
thoylindoles, naphthylmethylindoles, naphthoylpyrroles, naph-
thylmethylindenes and phenylacetylindoles (i.e. benzoylindoles)
(known as aminoalkylindoles or JWH compounds—after the name
of their inventor) (Huffman et al., 1994). Examples of naph-
thoylindoles include JWH-015, its n-pentyl homologue JWH-018
(Uchiyama et al., 2009; Chin et al., 1999), JWH-073 (an alkyl homo-
logue of JWH-018) (Aung et al., 2000), and JWH-398. An example
of phenylacetylindole is JWH-250, identified in “Spice” products in
Germany (EMCDDA, 2009a,b).
Both JWH-018 and CP-47,497-C8 produce cannabis-like effects.
Due to their high pharmacological potency in vitro, it is likely that
are active in relatively low doses. The duration of effects in humans,
compared to 9 -THC, seems to be shorter for JWH-018 (1–2 h) and
considerably longer for CP-47,497-C8 (5–6 h) (EMCDDA, 2009a,b).
3. Common synthetic cannabinoids found in “Spice”
products
3.1. JWH-018
JWH-018 is the first synthetic cannabinoid ever reported
through the Early Warning System (EWS) during 2008 (EMCDDA,
2008). JWH-018 is prominent component of several herbal prepara-
tions and is an effective CB1 receptor agonist that activates multiple
signalling pathways. Thus, it is likely that the subjective effects of
“Spice” are due to activation of cannabinoid CB1 receptors by JWH-
018, added to this herbal preparation (Mechoulam et al., 1988).
Despite the absence of clinical data in humans, some infor-
mation regarding the effects of some “Spice” products can be
provided through the Internet. Representative recorded data are
presented in an Internet chat room conversation, among “Spice”
users (http://www.bluelight.ru).
According to a user, 4–6 mg of JWH-073 was inhaled. The symp-
toms were “pressure behind the eyes and in the front of person’s
brain (similar experience and duration with cannabis), fuzziness
and mood improvement”. After smoking repetitively for a day, the
person had a cumulative effect much like smoking cannabis. By the
end of the day the person “was feeling tired, stoned and fuzzy”. The
person found the chemical as interesting as good cannabis.
Another user melted and smoked up 4 mg of JWH-018. The per-
son felt “as with cannabis and JWH-073, but with a slightly different
feeling in the brain”. The person experienced “waves of unusual
body energy”. The effects of JWH-018 were “a little more distinct
compared to cannabis”.
 I. Vardakou et al. / Toxicology Letters 197 (2010) 157–162
In another conversation, the user smoked 10 mg of JWH-018 in
the first hit, causing him/her anxiety. Nevertheless the user admit-
ted that he/she had experienced the highest drug “fix”.
A different user referred to a couple of small “hits” of JWH-018,
characterizing them as “strong stuff”.
Although toxicity studies are not conducted by an independent
research laboratory, it is reported that JWH-018, consider non-
toxic. The main observations by a Rat Repeat Dose Study were that
JWH-018 caused a severe lethargic, unresponsive catatonic state
at all doses tested from 0.1 to 10 mg/kg in rats. At the upper drug
concentration of 10 mg/kg there was decreased breathing and a
male rat died. It seemed the death was related to the catatonia and
decreased breathing rather than organ toxicity. A rat pharmacoki-
netic study was performed by bolus IV injection at 5 mg/kg and
blood was collected at various time points for 24 h post-dose. JWH-
018 showed a bi-phasic distribution suggesting both distribution
and elimination phases. The clearance was consistent of hepatic
blood flow rates in a rat of (55 ml/min/kg). The volume of distri-
bution suggests that the drug is well distributed. The half life is
∼2 h. No accumulation was observed (http://www.mindfully.org
(JWH-018 Tox Results Summary)).
3.2. CP-47,497
In the early 1980s, a group at Pfizer explored the development
of analgesics using the potent synthetic cannabinoid, (−)-9-nor-
9␤-hydroxyhexahydrocannabinol (HHC), as a template. This led
to the development of the CP-47,497 and CP-55,940 which were
found to be more potent than 9 -THC in vivo. However, none of
the CP-47,497 or CP-55,940 analogs have high affinity for the CB1
receptor or better than modest affinity for the CB2 receptor (Ottani
and Giuliani, 2001).
CP-47,497 is comparable or more potent than 9 -THC in anal-
gesic, motor depressant anticonvulsant, and hypothermic effects in
mice, rats, and dogs (Jiang et al., 2005). It also elicits vocalization
in palpated rats and ataxia in dogs. In drug discrimination studies
in rats, the stimulus properties of 9 -THC (3.2 mg/kg i.p.) are gen-
eralized to CP-47,497, with an absolute threshold dose 3–14 times
lower than the threshold dose of 9 -THC itself, depending on route.
Furthermore, rats were unable to discriminate between the stimu-
lus properties of equated i.p. doses of 9 -THC and CP-47,497 after
prolonged training. Despite its potent behavioral effects, CP-47,497,
like 9 -THC, does not resemble standard antipsychotic, antide-
pressant, antianxiety or hypnotic drugs in simple drug interaction
tests (Pertwee, 2005).
3.3. HU-210
HU-210, a synthetic agonist analog of 9 -THC, is a potent CB1
and CB2 receptor agonist. It binds to neuroblastoma cell mem-
brane CB1 receptors with about the same affinity as CP-55,940.
In mouse behavior models (hypothermia, analgesia, hypoactivity,
catalepsy), after s.c. and i.v. administration, the ED50 is 5–20 ␮g/kg
(http://www.caymanchem.com (HU-210: Product information)).
In 2007, Robinson et al. assessed the effects of this synthetic
cannabinoid on memory and hippocampal activity. The derived
data provide evidence that the underlying mechanism for the spa-
tial memory deficits induced by HU-210 in rats is the accompanied
abnormality in hippocampal cell firing. Single unit recordings from
principal neurons in hippocampal CA3 and CA1 confirmed HU-
210-induced attenuation of the overall firing activity lowering both
the number of complex spikes fired and the occurrence of bursts
(Robinson et al., 2007).
In 2008, Nabemoto proposed a mechanism for the HU-210
induced release of arachidonic acid. In this study, treatment with
HU-210 with greater concentrations than 3 ␮M caused the release
159
of arachidonic acid, an increase in [Ca2+ ] and translocation of
cPLA2 ␣ in PC12 cells in a dose-dependent manner probably via
cannabinoid CB1 receptor and/or CB1 -like receptor in neuronal
(Nabemoto et al., 2008).
In 2009, Dalton et al. reported a large dose-dependent CB1 recep-
tor adaptation after sub-chronic and chronic exposure to the potent
synthetic cannabinoid HU-210 that appears to be region specific
with development of tolerance to drug induced body weight loss.
The molecular mechanisms underlying tolerance to HU-210 seem
to be similar to those reported for 9 -THC (Dalton et al., 2009).
4. Tracking the World Wide Web and Patterns of use
Tracking the World Wide Web (www), it is clear that “Spice”
products are generally used by teenagers and young people, who
are interested in using psychotropic drugs. There are reports that
“users recommend using pipes” and that “the fumes of the burn-
ing mixture could be inhaled”. “Spice” products can be smoked,
sometimes together with cannabis or consumed orally as an infu-
sion (EMCDDA). Those smoking blends are so widespread that the
results were even found in a smoking blend comparison, for the
best cannabinoid smoking blends of 2009. 15.81% of 1822 partic-
ipants voted that “Spice diamond” is the best product among 41
different smoking blends. Another comparison was found about
the best synthetic cannabinoid. JWH-018 was voted by 31 partici-
pants as the best among 10 other synthetic cannabinoids. Recorded
data have been also found which make clear that advertisements
excite the users who care about legal drugs e.g. “I keep seeing these
chemical names popping up in thread titles and I am very intrigued.
Are these chemicals easily to obtain and legal?”. Users are inter-
ested to learn more details about “Spice” products e.g.1: “so what
exactly is this stuff? Anyone got any good sources of info on the
chems?” e.g. 2: “Apart from knowing that CP-47,497 is a cannabi-
noid I have no other info on this product. please enlighten me.”
(http://www.bluelight.ru).
As it has been reported in Germany, “Spice” products have been
consumed by cannabis users as a substitute, to enable them to pass
drug-screening tests. Especially young people interested in using
legal biogenic drugs are using “Spice” while sensation-seekers and
experimental drug users are attracted to “Spice” by media coverage.
“Spice” has been also reported in Sweden as a problem in prisons
and the probation service.
In 2009, new evidence about the dangers of “Spice” drugs at
the first International Psychonaut Web mapping has been released.
In this conference, two academics described the pharmacological
aspects of novel drugs of abuse and provided an overview of “Spice”
drugs as a result of a two-year European Commission-funded study,
to implement a regular monitoring of the www. According to the
preliminary results of this study, the drug is accessible to children
and young people, as there are no or very limited controls on any
of the websites selling the drug (http://www.p2002.sgul.ac.uk).
5. Case reports
A limited number of clinical reports clearly demonstrate signs
of addiction and withdrawal symptoms that were linked to the
chronic abuse of “Spice”, similar to syndromes observed with
cannabis abuse.
Case 1: Zimmermann et al. (2008), presented a case of a 20-
year-old patient who reported that for the last 8 months he had
been inhaling the smoke of “Spice Gold” initially 1 g daily.
The patient described that “Spice Gold” acted similarly to
cannabis but with “Spice Gold” experienced strong relaxation and
sedation. The main difference between “Spice” and cannabis was
that “Spice” produced less euphoria.
160 I. Vardakou et al. / Toxicology Letters 197 (2010) 157–162
Due to the tolerance, he had rapidly increased the dose to
a final dose of 3 g daily. Owing to the use of the substance, he
had often recently been listless and had difficulties in “thinking
clearly”. During a phase of abstinence he had developed symp-
toms of “profuse sweating” during the day and especially in the
night, as well as “internal unrest, tremor, palpitation, insomnia,
headache, diarrhea, nausea, and vomiting”. Additionally, he had
suddenly felt depressed and desperate. Symptoms had lasted for
2 days and had only abruptly disappeared after taking the drug
again. The patient requested medical treatment for “Spice Gold”
dependance.
Case 2: In 2010, Müller et al. presented the case of a 25-year-
old man who had smoked “Spice” on three different occasions (3 g
each).
Immediately after “Spice” use the patient had experienced
“imperative voices as well as recurrent paranoid hallucinations”.
Of interest, in both cases, urine screening revealed nega-
tive results. Additionally, both patients had history of psychotic
episodes at early age and positive family history of psy-
chosis.
It has been reported that patients with positive family his-
tory and thus a presumed genetic vulnerability are at higher risk
for developing drug associated psychosis (Fergusson et al., 2006;
Morgan and Curran, 2008).
Other cases: Some emergency cases were further reported in
several countries.
In Germany, effects on the cardiovascular and nervous system
like tachycardia and, in some cases, short-term loss of conscious-
ness, have been reported after “Spice” products use. A case of a
53-year-old woman has been also reported in Italy. Patient smoked
“Spice” and 1 h after use she was treated in an emergency room in
an excited state and in psychomotor agitation.
During 2008, the Swedish Poison Information Centre had 40
inquiries confirmed for “Spice” adverse effects.
In all of the above mentioned cases, the presence of the synthetic
cannabinoids was not confirmed by toxicological analysis.
Based on a few accounts from counselling centres in Austria,
the subjective effects of “Spice” were reported as highly variable,
ranging from mild to strong and it was reported that they could be
similar to cannabis or, alternatively, completely different. Finally,
similar reports from users in Romania suggested that “Spice” tasted
like herbal cannabis and the effects could be similar or even “better”
(EMCDDA Poison Control Centre, 2009).
Self-experiment: In 2008, Auwärter et al. decided to do a con-
trolled self-experiment with one type of herbal product in order to
prove pharmacological activity and to gain drug positive blood and
urine samples. One cigarette containing 0.3 g of “Spice Diamond”
was smoked and several blood and urine samples were collected.
Approximately 10 min post-use the first noticeable “cannabis-like”
effects occurred in the form of considerably reddened conjunctivae,
significant increase of pulse rate, xerostomia and a deterioration of
mood and perception. The subjects had the impression of being
moderately impaired. The effects continued for about 6 h under
slow attenuation. The whole next day, some minor after-effects
were still noticeable (2009).
6. Identification of synthetic cannabinoids in “Spice”
products
According to the EMCDDA Report on “Spice” extensive toxico-
logical investigations were undertaken in Germany and Austria
for the identification of “Spice” products ingredients. Chemical
analyses have revealed that the psychoactive effects of the herbal
mixture were due to added synthetic cannabinoids. Several ana-
lytical laboratories in Europe (Germany, Austria, Finland, France,
Hungary, Poland, Slovakia, Slovenia and the United Kingdom) have
confirmed the presence of synthetic cannabinoids in “Spice” prod-
ucts.
In 2008 a pharmaceutical company, THC-Pharm (Frankfurt, Ger-
many), had found JWH-018 to be present in some samples of “Spice”
products.
As it has already mentioned, through a self-report study and
chemical analysis of plant extracts and plasma samples after
consumption, a team of German scientists clearly identified the
presence of added synthetic compounds. Using an extract of “Spice
Diamond”, by combining the mass spectral information and the
results of nuclear magnetic resonance spectroscopy (NMR), the C8
homologue of the synthetic cannabinoid CP-47,497 was found to
be the main active component (Auwärter et al., 2009).
Furthermore, JWH-073 was identified in “Scope” product as well
as in other products in Germany, in Denmark, in the Netherlands
and in Finland (EMCDDA, 2009a,b).
In June 2009, HU-210 was identified for the first time in the
United Kingdom (EMCDDA, 2009a,b).
Recently, two new synthetic cannabinoids, the napthoylindole
JWH-398 and the phenylacetylindole JWH-250, have been iden-
tified in “Spice” products in the United Kingdom and Germany,
respectively (EMCDDA, 2009a,b).
In addition, Lindigkeit et al. (2009) managed to isolate and
quantify the psychoactive compounds added to herbal blends. The
amount of added cannabimimetics varied over one order of mag-
nitude and ranged from 2.3 mg/g up to 22.9 mg/g. The range is in
accordance to the estimated 1% of CP-47,497-C8 reported for “Spice
Diamond”.
One batch of samples, which was acquired before German pro-
hibition, found to contain CP-47,497-C8 (together with its isomeric
by-product) or JWH-018. Another batch of samples, which was
acquired after German prohibition, showed a more complex pat-
tern. While CP-47,497-C8 can be still found on the market, it was
reported the first appearance of JWH-073 (C4-homologue) as alter-
native for JWH-018.
This research proves that just 4 weeks after the prohibition
a second generation products were flooding the market. The
speed of introduction of new products and the use of JWH-073
as a substitute for JWH-018 not only show that the producers
are well aware of the legal frameworks, but that they already
have an array of replacement products on hand (Lindigkeit et al.,
2009).
In 2010 Uchiyama et al. analysed herbal products which were
obtained from the Japanese market. Forty-six herbal products cur-
rently being sold in Japan for their expected cannabis-like effects
were purchased via the Internet. These products appeared in pri-
marily two forms i.e., as bits of dried leaves or as cigarettes.
GC-MS and LC–MS analyses indicated that most of the products
contained cannabicyclohexanol and JWH-018. Cannabicyclohex-
anol is a cannabimimetic agent that produces more potent effects
than those of typical cannabinoids. Cis- and trans-diastereomer of
cannabicyclohexanol was also detected. Research revealed also that
these “Spice” products contained CP-47,497 and JWH-073.
Only recently, Teske et al. (in press) presented a validated
method for the detection and quantification of naphthalen-1-yl-(1-
pentylindol-3-yl)methanone (JWH-018), by means of LC–MS/MS
in serum while Sobolevsky et al. (in press), managed to identify
two main monohydroxylated metabolites of JWH-018 in forensic
urine samples (n = 3). As reported, the behavioural effects were
similar to those typical of the administration of the marijuana
products, such as the reddening of eyes, tachycardia, anxiety,
paranoia and hallucinations accompanied by a short-term mem-
ory defects and the impaired sense of time. Using gas and liquid
chromatography combined with tandem mass spectrometry the
authors identified the main metabolites but not the parent com-
pound.
 I. Vardakou et al. / Toxicology Letters 197 (2010) 157–162
7. Legal status
Products containing HU-210 and similar cannabinoids are
controlled in a number of countries, including Austria (3rd
March 2009), Germany (22nd January 2009), the Nether-
lands, Switzerland, Canada (Intelligence alert, 2009) and UK
(23rd December 2009) (http://drugs.homeoffice.gov.uk/news-
events/latest-news/control-of-legal-highs-dec09.html).
Furthermore, responding to potential health concerns, France
(24th February 2009), Hungary (9th March 2009), Poland (8th May
2009), Luxembourg (4th May 2009), Lithuania (27th May 2009),
Estonia (24th July 2009) and Sweden (15th September 2009) have
recently taken legal actions to ban or otherwise control “Spice”
products and related compounds (EMCDDA, 2009a,b). The Advisory
Council on the Misuse of Drugs (ACMD) advised the United King-
dom government to classify and control “spice” products (ACMD,
2009). Following the prohibition of JWH-018 in several countries
during 2009, the smoking mixtures containing this compound were
legal in Russia until the middle of January 2010 (Sobolevsky et al.,
in press).
None of the listed ingredients of “Spice” are controlled in
most of the United States. Kansas is the only state that has
banned spice (March 2010) while several other states are consid-
ering some sort of regulatory legislation (http://news.hjnews.com/
news/article 76514f64-3aaf-11df-afe1-01cc4c002e0.html). On the
other hand U.S. Military Rules prohibit “Spice”. Thus U.S. Army
has instituted a policy banning “Spice” (http://www.army.mil
(Official homepage of the United States Army)). U.S. Air Force
has prohibited “Spice” (http://www.af.mil (The official website
of the U. S. Air Force)). U.S. Marines have banned “Spice” and
the component chemicals entirely (http://www.marines.mil (Offi-
cial website of the United States Marine Corps)), while “Spice”
products are considered completely prohibited in the U.S. Navy
(http://www.navy.mil (Official website of the United States Navy)).
8. Discussion
“Spice” products contain synthetic cannabinoids that bind to
cannabinnoid-like receptors and they are stronger than natural
cannabis. These cannabinoids can be considered as new “designer
drugs”.
It remains unclear where and how the actual production of the
herbal mixtures, synthetic cannabinoids and their addition to the
herbal mixture takes place. It can be assumed that the producers
buy cannabinoids from laboratories in Asia (e.g. China) or other
countries that offer cheap raw materials, dissolve the compounds in
a solvent, spray the solution on the plant and evaporate the solvent
before packaging the herbal mixtures. It is evident that the produc-
ers are purposely risk the health of consumers to skim high profits.
At a price of 20–30 D (∼$25–40) for a 3-g packet, the incense is sig-
nificantly more expensive than marijuana (5–7 D /g). Nevertheless,
its popularity has spread via Internet. It is possible that dealers have
a base worldwide and from there, they send “Spice” from one coun-
try to the other. As a result it is very difficult for national authorities
to trace back the whole thing (Piggee, 2009).
Apart from the lack of information regarding the origin of these
compounds, the main problem focuses on the ineffective identifi-
cation procedures. Even though the identification of few synthetic
cannabinoids in a number of samples was fulfilled, the detection
of the whole range of all related substances is not yet feasible.
The laboratories cannot identify the unknown compounds quickly
because currently these compounds are not included in any mass
or UV spectra library. However, reference substances for quanti-
tative and qualitative investigations are rare; in Germany at the
time of the current experiment only JWH-018 was commercially
161
available (Teske et al., in press). Furthermore, almost daily new
products which declared as herbal incense, with different names
and different packaging are appeared, while some products do not
contain any pharmacologically active compounds misguiding by
that way the researchers. Finally, the extracts of the mixtures ren-
dered very complex matrices which make their analysis extremely
difficult and high amounts of non-psychoactive compounds, such
as vitamin E, “mask” the active components (EMCDDA, 2009a,b).
Of interest, genetic analysis results of some herbal products did
not match with the plant series named in the labels, providing the
potentiality that the plant materials are included mainly as diluents
for the synthetic compounds (Uchiyama et al., 2010).
It seems that the producers are moving on to the next prod-
uct, always one step ahead of the law contributing to the already
existing problem for the identification of these compounds.
Since the synthetic cannabinoids of “Spice” products have not
tested in humans, the health risk of the inhaled smoke is unknown.
In the case of JWH-018, it can be presumed that, according to
its structure, there is a potential carcinogenic effect. In addition,
accidental overdosing may lead to a risk of severe psychiatric com-
plications and life-threatening conditions in the case of full CB
receptor agonist (versus THC which acts as partial agonist). Finally,
tolerance to these products may be developed relatively rapid
which might be associated with dependence.
Thus, considering the limited information about the origin of
these substances in combination with the absence of reference
standards and the potential health risks, the alertness of research
community is of great importance in order to track and prevent the
spread of “Spice” products.
9. Conclusion
This article focuses at all available data regarding the mode
of action, the existing identification problems and the current
worldwide legislation status of synthetic cannabinoids in “Spice”
products.
Even though “Spice” products are commercialized only as
incense for their rich aroma, the users utilize them exactly like
cannabis by inhaling the smoke. “Spice” products are designed for
youngsters that are willing to try cannabis but are afraid of the judi-
cial consequences and/or the reputation of the narcotics scene. The
trap is that these products might be the first step towards drugs of
abuse.
Although clinical reports clearly demonstrate signs of depen-
dence and withdrawal symptoms that were linked to the chronic
abuse of “Spice” and were similar to syndromes observed in
cannabis abuse, these products are still advertised in a very sys-
tematic way to manipulate young people and their large demand
for “legal-drugs”.
Research community must remain vigilant, as various new
“Spice” products continuously appear. This, together with the vari-
ety and number of synthetic cannabinoids that could be potentially
added to herbal products, continues to pose challenges for identi-
fication, monitoring and risk assessment of this new trend.
Conflict of interest statement
The authors declare that there are no conflicts of interest.
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