Abstract
We have combined morphological, molecular, and chemical techniques in order to identify the plant and chemical composition of some last-generation smart drugs, present on the market under the following names: Jungle Mistic Incense, B-52, Blendz, and Kratom 10x. Micromorphological analyses of botanical fragments allowed identification of epidermal cells, stomata, trichomes, starch, crystals, and pollen. DNA barcoding was carried out by the plastidial gene rbcL and the spacer trnH-psbA as universal markers. The combination of morphological and molecular data revealed a mixture of plants from different families, including aromatic species, viz., Lamiaceae and Turneraceae. GC-MS and LC-MS analyses on ethanol or methanol extracts showed the presence of synthetic cannabinoids, including JWH-250 in Jungle, JWH-122 in B-52, and JWH-073 and JWH-018 in Blendz. In Kratom 10x, only the indole alkaloid mitragynine was detected. All the identified synthetic cannabinoids, apart from mitragynine, are under the restriction of law in Italy (TU 309/90). Synthetic cannabinoid crystals were also identified by scanning electron microscopy and energy dispersive X-ray spectroscopy, which also detected other foreign organic chemicals, probably preservatives or antimycotics. In Kratom only leaf fragments from Mitragyna speciosa, containing the alkaloid mitragynine, were found. In the remaining products, aromatic plant species have mainly the role of hiding synthetic cannabinoids, thus acting as a “green shuttle” rather than as real drugs. Such a multidisciplinary approach is proposed as a method for the identification of herbal blends of uncertain composition, which are widely marketed in “headshops” and on the Internet, and represent a serious hazard to public health.
Similar content being viewed by others
References
Lindigkeit R, Boehme A, Eiserloh I, Luebbecke M, Wiggermann M, Ernst L, Beuerle T (2009) Spice: a never ending story? Forensic Sci Int 191:58–63
Schmidt MM, Sharma A, Schifano F, Feinmann C (2011) “Legal highs” on the net—evaluation of UK-based websites, products and product information. Forensic Sci Int 206:92–97
Atwood BK, Lee D, Straiker A, Widlanski TS, Mackie K (2011) CP47,497-C8 and JWH073, commonly found in ‘Spice’ herbal blends, are potent and efficacious CB(1) cannabinoid receptor agonists. Eur J Pharmacol 659:139–45
Dresen S, Ferreirós N, Pütz M, Westphal F, Zimmermann R, Auwärter V (2010) Monitoring of herbal mixtures potentially containing synthetic cannabinoids as psychoactive compounds. J Mass Spectrom 45:1186–94
Auwärter V, Dresen S, Weinmann W, Müller M, Pütz M, Ferreirós N (2009) ‘Spice’ and other herbal blends: harmless incense or cannabinoid designer drugs? J Mass Spectrom 44:832–837
Vardakou I, Pistos C, Spiliopoulou C (2010) Spice drugs as a new trend: mode of action, identification and legislation. Toxicol Lett 197:157–162
www.barcoding.si.edu. Accessed 22 Jun 2012
Hebert PDN, Cywinska A, Ball SL, deWaard JR (2003) Biological identifications through DNA barcodes. Proc R Soc Lond B 270:313–321
Casiraghi M, Labra M, Ferri E, Galimberti A, De Mattia F (2010) DNA barcoding: a six-question tour to improve users' awareness about the method. Brief Bioinform 11:440–453
Fay MF, Bayer C, Alverson WS, de Bruijn AY, Chase MW (1998) Plastid rbcL sequence data indicate a close affinity between Diegodendron and Bixa. Taxon 47:43–50
Newmaster SG, Ragupathy S (2009) Testing plant barcoding in a sister species complex of pantropical Acacia (Mimosoideae, Fabaceae). Mol Ecol Resour 9:172–180
Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948. doi:10.1093/bioinformatics/btm404
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410
BOLD Identification System (IDS) http://www.boldsystems.org/index.php/IDS_OpenIdEngine. Accessed 12 Oct 2012
Jackson BP, Snowdon DW (1990) Atlas of microscopy of medicinal plants, culinary herbs and spices. CRC, Boca Raton
Rahfeld B (2011) Mikroskopischer farbatlas pflanzlicher drogen. Spektrum Akademischer Verlag, Plankstadt
Tschirch A, Oesterle O (1900) Anatomischer atlas der pharmakognosie und nahrungsmittelkunde. Leipzig Chr. Herm. Tauchnitz, Bern, Schweiz
Schneider A (1902) Powdered vegetable drugs. Calumet, Pittsburgh
Kumar S, Taneja R, Sharma A (2006) Pharmacognostic standardization of Turnera aphrodisiaca Ward. J Med Food 9:254–260
Buchner R, Weber M (2000 onwards) Descriptions, illustrations, identifications, and information retrieval. PalDat-a palinologicaldatabase: http://www.paldat.org/. Accessed 12 Oct 2012
Erdtman G (1952) Pollen morphology and plant taxonomy I. Almqvist & Wiksell, Stockholm
Faegri K, Iversen J (1975) Textbook of pollen analysis. Hafner, New York
Punt W et al. (1976–2003) The Northwest European pollen flora (NEPF). Vols 1–8. Elsevier, Amsterdam
Lersten NR, Horner HT (2011) Unique calcium oxalate “duplex” and “concretion” idioblasts in leaves of tribe Naucleeae (Rubiaceae). Am J Bot 98:1–11
Jensen WA (1962) Botanical histochemistry. Freeman, San Francisco
Martin PS, Drew CM (1969) Scanning electron photomicrographs of southwestern pollen grains. J Ariz-Nev Acad Sci 5:147–176
Martin PS, Drew CM (1970) Additional scanning electron micrographs of southwestern pollen grains. J Ariz-Nev Acad Sci 6:140–161
Pathan AK, Bond J, Gaskin RE (2008) Sample preparation for scanning electron microscopy of plant surfaces—horses for courses. Micron 39:1049–1061
Solomon AM, King JE, Martin PS, Thomas J (1973) Further scanning electron photomicrographs of southwestern pollen grains. J Ariz-Nev Acad Sci 8:135–157
Zafar M, Khan MA, Ahmad M, Sultana S, Qureshi R, Tareen RB (2010) Authentication of misidentified crude herbal drugs marketed in Pakistan. Journal of Medicinal Plants Research 4:1584–1593 Available online at http://www.academicjournals.org/JMPR ISSN 1996–0875
Uchiyama N, Kawamura M, Kikura-Hanajiri R, Goda Y (2011) Identification and quantitation of two cannabimimetic phenylacetylindoles JWH-251 and JWH-250, and four cannabimimetic naphthoylindoles JWH-081, JWH-015, JWH-200, and JWH-073 as designer drugs in illegal products. Forensic Toxicol 29:25–37
Department for Antidrug Policies (DPA) National early warning system (N.E.W.S.). www.allertadroga.it/sapd/login/. Accessed 20 Dec 2012
Scientific Working Group for the Analysis of Seized Drugs. http://www.swgdrug.org/ms.htm. Accessed 20 Dec 2012
Walker MD, Ahmad SJ (1970) The Mitragyna species of Asia. XVII. The anatomy of the leaves of Mitragyna javanica var. microphylla. Planta Medica 18:55–65
Razafimandimbison SG, Bremer B (2002) Phylogeny and classification of Naucleeae s.l. (Rubiaceae) inferred from molecular (ITS, rbcL, and trnT-F) and morphological data. Am J Bot 89:1027–1041
Kennedy PD, Collin W (2010) Automated ion trap screening method for the detection of synthetic cannabinoids in commercial herbal incense products. Synthetic cannabinoid screening. https://www.caymanchem.com/app/screen/Static/restrict/public/path/sampleApplications,CaymanChemicalSyntheticCannabinoidScreening.pdf/filename/CaymanChemicalSyntheticCannabinoidScreening.pdf. Accessed 22 June 2012
Decreto del Presidente della Repubblica 9 ottobre 1990, n. 309 GU n. 62 del 15-3-2006 - Suppl Ordinario n.62
United Nations Office on Drugs and Crime (UNODC) (2009) Recommended methods for the identification and analysis of cannabis and cannabis products. United Nations New York. http://www.unodc.org/documents/scientific/ST-NAR-40-Ebook.pdf. Accessed 26 Jan 2012
Uchiyama N, Kikura-Hanajiri R, Kawahara N, Goda Y (2009) Identification of a cannabimimetic indole as a designer drug in a herbal product. Forensic Toxicol 27:61–66
Dargan PI, Hudson S, Ramsey J, Wood DM (2011) The impact of changes in UK classification of the synthetic cannabinoid receptor agonists in ‘Spice’. Int J Drug Policy 22:274–277
Gottardo R, Chiarini A, Dal Prà I, Seri C, Rimondo C, Serpelloni G, Armato U, Tagliaro F (2012) Direct screening of herbal blends for new synthetic cannabinoids by MALDI-TOF MS. J Mass Spectrom 47:141–146
Uchiyama N, Kikura-Hanajiri R, Ogata J, Goda Y (2010) Chemical analysis of synthetic cannabinoids as designer drugs in herbal products. Forensic Sci Int 198:31–38
Kumar S, Taneja R, Sharma A (2005) The genus Turnera: a review update. Pharm Biol 43:383–391
Ministero della Salute. D.L. 16/6/2010 http://www.politicheantidroga.it/media/336190/d.m._16_giugno_2010.pdf. Accessed 20 Jun 2011
Ministero della Salute. D.L. 11/5/2011 http://www.politicheantidroga.it/media/446920/d.m%2011%20maggio%2011.pdf. Accessed 8 Jul 2011
Schultes RE, Hofmann A, Rätsch C (2001) Plants of the gods. Healing Arts Press, Rochester, Vermont
De Mattia F, Bruni I, Galimberti A, Cattaneo F, Casiraghi M, Labra M (2011) A comparative study of different DNA barcoding markers for the identification of some members of Lamiaceae. Food Rev Int 44:693–702
De Mattia F, Gentili R, Bruni I, Galimberti A, Sgorbati S, Casiraghi M, Labra M (2012) A multi-marker DNA barcoding approach to save time and resources in vegetation surveys. Bot J Linn Soc 169:518–529
Serrano R, da Silva G, Silva O (2010) Application of light and scanning electron microscopy in the identification of herbal medicines. In Méndez-Vilas A, Díaz J (eds) Microscopy: science, technology, applications and education. ©FORMATEX 2010, Badajoz, Spain, pp.182–190
Acknowledgments
We are very grateful to Laura Negretti (DISTAV Università di Genova) for the technical assistance in SEM–EDS analyses, and to Neil Campbell (Università di Milano-Bicocca) for language revision. We also thank Riccardo Albericci, Curator of the Botanical Garden Clelia Durazzo Grimaldi of Genoa, Italy, and the firm A. Minardi & Figli s.r.l. (Bagnocavallo, Ravenna, Italy) for providing plant samples and herbal blends used for comparison. This research was partially supported by Fondazione Carige, Genoa, Italy (nr. 2013.0132-2).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Cornara, L., Borghesi, B., Canali, C. et al. Smart drugs: green shuttle or real drug?. Int J Legal Med 127, 1109–1123 (2013). https://doi.org/10.1007/s00414-013-0893-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00414-013-0893-9