COMMUNICATION www.rsc.org/methods | Analytical Methods

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Spice up your life: screening the illegal components of ‘Spice’ herbal products

Claire E. J. Emanuel, Bill Ellison and Craig E. Banks*

Received 29th March 2010, Accepted 14th April 2010

First published as an Advance Article on the web 4th May 2010

DOI: 10.1039/c0ay00200c

‘Spice’ is a herbal blend which was freely available in the UK until

December 2009 after which it was classified as a Class B drug. The

Spice product range includes ‘Gold’ and ‘Diamond’ and a very

limited number of reports have identified that these contain non-

traditional cannabinoids. We have determined for the first time the

components of Spice ‘Gold Spirit’ using Gas Chromatography-

Mass Spectroscopy and also explored a potential screening

approach using Solid Probe Mass Spectroscopy which requires no

liquid–liquid extraction. This methodology has the potential to

facilitate the accelerated screening of illegal components in ‘Spice’

and other related herbal-type products.

Introduction

Spice is a herbal blend sold predominantly in the European

community and is well known by users to have effects similar to that

of Cannabis.1 Spice was declared as a ‘bio-drug’ since the listed

ingredients indicate bioactive herbs which contain alkaloids and

generally produce cannabis-like effects.1–3 Accordingly the popularity

of these products dramatically increased and was given the tag line of

‘legal drugs’.

In 2008 THC Pharma reported the active ingredients of Spice to be

‘JWH-018’ (see Scheme 1) which is a cannabinoid agonist from the

aminoalkylindole family which has been shown to have a binding

affinity for CB1 receptors at low nanomolar levels (�9 nM). Note

that this is the same cannabinoid receptor that is linked to behaviours

affected by tetrahydrocannabinol. Very recently reports by Atwood

et al. have demonstrated that JWH-018 is a potent and effective CB1

receptor agonist that activates multiple CB1 receptor signalling

pathways.4 Thus it is highly likely that the psycho-activity of ‘Spice’ is

due to the JWH-018 agonist.4

Following the report by THC Pharma, Auwarter et al.1 and

Uchiyama et al.5 identified and characterized the CP 47,497-C8 (see

Scheme 1) homolog and its isomer, a synthetic by-product, in Spice

Silver, Gold and Diamond as well as ‘Yucatan Fire’ and ‘Sence’.

Note that it has been reported that the analgesic potency of CP

47,497-C8 is 5 to 10 times higher compared with tetrahydrocannab-

inol.6 Lindigkeit and co-workers2 have analysed Spice Gold in

Germany with mass spectrometry and found that the samples con-

tained CP 47,497-C8 and JWH-018 but after the German Health

Authorities prohibited the active components of Spice on January

22nd 2009, JWH-018 was found to be absent in Spice products.2

Faculty of Science and Engineering, School of Biology, Chemistry andHealth Science, Division of Chemistry and Materials, ManchesterMetropolitan University, Chester Street, Manchester, M1 5GD, Lancs,UK. E-mail: c.banks@mmu.ac.uk; Fax: +44 (0)1612476831; Tel: +44(0)1612471196

614 | Anal. Methods, 2010, 2, 614–616

However, a new analogue JWH-073 was found.2 In the UK, Spice

was legal until December 2009 where the synthetic cannabinoids,

which are sprayed onto herbal smoking products, are now classified

as a Class B drug.7,8 Given that the manufacturers can readily change

the components of Spice, a methodology that can readily identify the

presence of prohibited compounds in these complex mixtures would

be highly welcome.

In this communication we explore for the first time the components

of the Spice product ‘Gold Spirit’ via Gas Chromatography-Mass

Spectrometry and explore the possibility of using Solid Probe Mass

Spectrometry which precludes the need for liquid–liquid extraction.

Experimental section

The Gas Chromatography-Mass Spectroscopy was a Hewlett

Packard 5890 Series 2 Gas Chromatogram. This machine is equipped

with a 30 m� 0.25 mm film¼ 0.25 mm Restek column (5% diphenyl–

95% dimethyl polysiloxane) with helium as the carrier gas (flow rate

1 ml min�1). Temperature program: 50 �C (2 min) to 12 �C min�1 to

290 �C (6 min). The GC was coupled directly to a Hewlett Packard

5972 series mass spectrometer with an electron ionization mode at

70 eV. The acquisition rate was 1.7 scans per second with a mass

range of 50–500 amu, using MSDChemstation software.

A Finnigan trace mass spectrometer coupled with a trace gas

chromatogram 2000 series was also used which is equipped with a

J & W Scientific 30 m � 25 mm film ¼ 0.25 mm analytical column

and helium gas carrier (flow rate 1.0 ml min�1). Temperature

program: 50 �C (2 min) to 12 �C min�1 to 285 �C (15 min). Mass spec.

parameters electron ionization mode at 70 eV using Excalibur soft-

ware. The emission current was 300 micro-amps, detector voltage

500 �C, the source temperature 280 �C. The acquisition rate was

Scheme 1 Structures of compounds commonly found in Spice herbal

products. (1) CP 47,497-C8; (2) CP 47,497-C8 isomer and (3) JWH-018.

This journal is ª The Royal Society of Chemistry 2010

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2.5 scans per second, mass range 45–460 amu. The Solid Probe Mass

Spectrometer uses a DIP (direct insertion probe) equipped with a vial

containing a small amount of sample was inserted into the mass

spectrometer. Analysis settings: detector voltage 500.0 V, source

temperature 250 �C the interface temperature 200 �C, emission

current 300 mV. The temperature was set at 30 �C for 1 min followed

by a temperature ramp of 800 �C min�1 taking the temperature to 400�C. The temperature was then held at 750 �C for 2 min. The acqui-

sition type: full scan at 1.8 scans per second scan range 50–640 amu.

Mass spectra were obtained and compared with recent literature

reports.1,2

Spice ‘Gold’, ‘Diamond’ and ‘Gold Spirit’ were purchased from

the Internet in 2009. The ingredients listed by the supplier for Spice

Gold are: bay bean, blue lotus, Lion’s Tail, Indian Warrior, Dwarf

skullcap, Maconha brava, Pink Lotus, Marshmallow, Red Clover,

Rose, Siberian motherwort, Vanilla and honey. Ingredients listed for

Spice Gold Spirit are: Leonurus, Cardiaca, Pedicularis, Canadensis,

Scutellaria, Latero flora, Althaea officinalis, Rosa damascena, Vanilla

planifolia. Finally, the ingredients listed for Spice Diamond are: Bay

bean, Blue Lotus, Lion’s Tail, Indian Warrior, Dwarf skullcap,

Maconha brava, Pink Lotus, Marshmallow, Red Clover, Rose,

Siberian motherwort, vanilla, honey, aroma.

A liquid–liquid extraction of the chosen Spice product was

undertaken with ethanol with the sample placed into a laboratory

ultrasonic bath for 15 minutes. The sample was then filtered using

a Buchner filter. The solution was diluted further with the addition of

ethanol.

Results and discussion

The GC-MS chromatograms of the three Spice products are depicted

in Fig. 1. The spectra obtained from Spice Gold reveal the presence of

ethyl vanillin, a-tocopherol and g-tocopherol. Also found present

Fig. 1 GC-MS chromatograms for various Spice products.

This journal is ª The Royal Society of Chemistry 2010

was CP 47,497-C8 which has been reported to be a cannabinoid

receptor agonist.1 These constituents of Spice Gold compare well

with previous literature reports.1,2 In the case of Spice Diamond,

caffeine, a-tocopherol and g-tocopherol and palmitic acid are found

to be present with the latter being saturated fatty acids commonly

found in plants and given the herbal origin of Spice products its

presence is expected. Interestingly CP 47,497-C8 is found to be

present which has been observed before in Spice Silver, Gold and

Diamond1 with its analgesic potency reported to be 5 to 10 times

higher compared with tetrahydrocannabinol.6 We also find that

JWH-018 (341, 324, 284, 214 m/z) is present in Spice Diamond which

has only been reported once before by Auwarter et al.1 Recently

Atwood et al.4 have proven that JWH-018 is a potent and effective

CB1 receptor agonist that activates multiple CB1 receptor signalling

pathways.4

Next we turn to exploring the components of Spice Gold Spirit

which is a new addition to the Spice herbal range and until now, its

components have never been quantified. Fig. 1 depicts a typical

chromatogram where a-tocopherol and JWH-018 are found to be

present. Clearly this new range of Spice has the highly active

cannabinoid mimic which has attracted attention from consumers as

a legal high. A recent study by Lindigkeit et al.2 has demonstrated

that Spice Gold purchased before the prohibition by German

Authorities contained CP 47,497-C8 and JWH-018 but samples

analysed after the prohibition were found not to contain the highly

active JWH-018 compound. Our work has shown that a simple

liquid–liquid extraction followed by GC-MS can quantify, without

doubt, the presence of recently illegal compounds. Given that the

manufacturers of Spice herbal blends are continuously changing the

compounds,2 there is a need for the quantification of these banned

substances, for example, in a Forensic situation. Consequently we

turn to exploring the concept of using Solid Probe Mass Spectrom-

etry for quantifying the components of Spice herbal products and

associated herbal type products.

Fig. 2A displays a typical spectra obtained using the Solid Probe

MS where it is evident that the fragmentation pattern of a-tocopherol

(430, 205, 165 m/z) and CP 47,497-C8 (332, 314, 233, 215 m/z) is

Fig. 2 Mass spectra obtained using the Solid Probe MS before (A) and

after (B) subtraction for Spice Gold.

Anal. Methods, 2010, 2, 614–616 | 615

Fig. 3 Mass spectra obtained using the Solid Probe MS before (A) and

after (B) subtraction for Spice Gold Spirit.

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present. The Solid Probe MS also functions separately as a stand

alone GC-MS and using the chromatograph obtained in Fig. 1 the

mass spectra resulting from a-tocopherol were determined and sub-

tracted from that of the spectra in Fig. 2A. The resulting spectrum is

shown in Fig. 2B which allows the fragmentation pattern of CP

47,497-C8 to be clearly visible and quantifiable. Note that it is entirely

reasonable to subtract the spectra of the usually obtained a-tocoph-

erol which is almost entirely found in Spice herbal products. Last we

616 | Anal. Methods, 2010, 2, 614–616

consider the case of Spice Gold Sprit. Fig. 3 depicts the mass spectra

before and after subtraction from a-tocopherol with the latter

allowing the quantification of JWH-018 (341, 324, 284, 214), the

potent and effective CB1 receptor agonist that is currently banned in

a plethora of countries.

Conclusions

We have quantified via GC-MS for the first time the components of

the herbal product Spice ‘Gold Spirit’ which is found to contain the

recently banned compounds JWH-018 and CP 47,497-C8. Addi-

tionally we have shown that Solid Probe Mass Spectrometry can be

used precluding the need for liquid–liquid extraction allowing the fast

screening of Spice herbal products. This screening methodology is

useful for the rapid analysis of any possible prohibited substances

which should be followed up fully with quantification via GC-MS.

References

1 V. Auwarter, S. Dresen, W. Weinmann, M. Muller, M. Putz andN. Ferreiros, J. Mass Spectrom., 2009, 44, 832.

2 R. Lindigkeit, A. Boehme, I. Eiserloh, M. Luebbecke andM. Wiggermann, Forensic Sci. Int., 2009, 191, 58.

3 M. A. Neukamm, T. E. Muerdter, C. Knabbe, H.-D. Wehner andF. Wehner, Blutalkohol, 2009, 46, 373.

4 B. K. Atwood, J. Huffman, A. Straiker and K. Mackie, Br. J.Pharmacol., 2010, DOI: 10.1111/j.1476-5381.2009.00582.x.

5 N. Uchiyama, R. Kikura-Hanajiri, N. Kawahara, Y. Haishima andY. Goda, Chem. Pharm. Bull., 2009, 57, 439.

6 B. K. Koe, G. M. Milne, A. Weissman, M. R. Johnson andL. S. Melvin, Eur. J. Pharmacol., 1985, 109, 201.

7 http://drugs.homeoffice.gov.uk/publication-search/acmd/acmd-report-agonists.html.

8 http://news.bbc.co.uk/1/hi/uk/8427439.stm.

This journal is ª The Royal Society of Chemistry 2010