A Metabolomic Approach Applied to a Liquid Chromatography Coupled to High-Resolution Tandem Mass Spectrometry Method (HPLC-ESI-HRMS/MS): Towards the Comprehensive Evaluation of the Chemical Composition of Cannabis Medicinal Extracts.

INTRODUCTION Cannabis sativa L. is a powerful medicinal plant and its use has recently increased for the treatment of several pathologies. Nonetheless, side effects, like dizziness and hallucinations, and long-term effects concerning memory and cognition, can occur. Most alarming is the lack of a standardised procedure to extract medicinal cannabis. Indeed, each galenical preparation has an unknown chemical composition in terms of cannabinoids and other active principles that depends on the extraction procedure. OBJECTIVE This study aims to highlight the main differences in the chemical composition of Bediol® extracts when the extraction is carried out with either ethyl alcohol or olive oil for various times (0, 60, 120 and 180 min for ethyl alcohol, and 0, 60, 90 and 120 min for olive oil). METHODOLOGY Cannabis medicinal extracts (CMEs) were analysed by liquid chromatography coupled to high-resolution tandem mass spectrometry (LC-MS/MS) using an untargeted metabolomics approach. The data sets were processed by unsupervised multivariate analysis. RESULTS Our results suggested that the main difference lies in the ratio of acid to decarboxylated cannabinoids, which dramatically influences the pharmacological activity of CMEs. Minor cannabinoids, alkaloids, and amino acids contributing to this difference are also discussed. The main cannabinoids were quantified in each extract applying a recently validated LC-MS and LC-UV method. CONCLUSIONS Notwithstanding the use of a standardised starting plant material, great changes are caused by different extraction procedures. The metabolomics approach is a useful tool for the evaluation of the chemical composition of cannabis extracts. Copyright © 2017 John Wiley & Sons, Ltd.

[1]  M. Cascio,et al.  Cannabidiolic acid prevents vomiting in Suncus murinus and nausea‐induced behaviour in rats by enhancing 5‐HT1A receptor activation , 2013, British journal of pharmacology.

[2]  Y. Shoyama,et al.  Tetrahydrocannabinolic acid, a genuine substance of tetrahydrocannabinol. , 1967, Chemical & pharmaceutical bulletin.

[3]  B. Colasanti.  A comparison of the ocular and central effects of delta 9-tetrahydrocannabinol and cannabigerol. , 1990, Journal of ocular pharmacology.

[4]  Natalie I. Tasman,et al.  A Cross-platform Toolkit for Mass Spectrometry and Proteomics , 2012, Nature Biotechnology.

[5]  G. Gigli,et al.  Analytical and preparative enantioseparation and main chiroptical properties of Iridium(III) bis(4,6-difluorophenylpyridinato)picolinato. , 2016, Journal of chromatography. A.

[6]  G. Moreno-Sanz.  Can You Pass the Acid Test? Critical Review and Novel Therapeutic Perspectives of Δ9-Tetrahydrocannabinolic Acid A , 2016, Cannabis and cannabinoid research.

[7]  W. Vaes,et al.  Determination of cannabinoids in cannabis products using liquid chromatography-ion trap mass spectrometry. , 2004, Journal of chromatography. A.

[8]  Arno Hazekamp,et al.  Cannabis Oil: chemical evaluation of an upcoming cannabis-based medicine , 2013 .

[9]  M. Zoli,et al.  Simultaneous measurement of adenosine, dopamine, acetylcholine and 5-hydroxytryptamine in cerebral mice microdialysis samples by LC-ESI-MS/MS. , 2012, Journal of pharmaceutical and biomedical analysis.

[10]  J. Bronstein,et al.  Systematic review: Efficacy and safety of medical marijuana in selected neurologic disorders , 2014, Neurology.

[11]  K. Franson,et al.  The Pharmacologic and Clinical Effects of Medical Cannabis , 2013, Pharmacotherapy.

[12]  B. Colasanti.  A Comparison of the Ocular and Central Effects of Δ9-Tetrahydrocannabinol and Cannabigerol , 1990 .

[13]  Gary Siuzdak,et al.  Liquid chromatography quadrupole time-of-flight mass spectrometry characterization of metabolites guided by the METLIN database , 2013, Nature Protocols.

[14]  Wieland Peschel,et al.  ¹H NMR and HPLC/DAD for Cannabis sativa L. chemotype distinction, extract profiling and specification. , 2015, Talanta.

[15]  Y. Choi,et al.  Metabolic fingerprinting of Cannabis sativa L., cannabinoids and terpenoids for chemotaxonomic and drug standardization purposes. , 2010, Phytochemistry.

[16]  Giuseppe Cannazza,et al.  Medicinal cannabis: Principal cannabinoids concentration and their stability evaluated by a high performance liquid chromatography coupled to diode array and quadrupole time of flight mass spectrometry method. , 2016, Journal of pharmaceutical and biomedical analysis.

[17]  W. Weinmann,et al.  Simultaneous quantification of delta-9-THC, THC-acid A, CBN and CBD in seized drugs using HPLC-DAD. , 2014, Forensic science international.

[18]  Bharathi Avula,et al.  Decarboxylation Study of Acidic Cannabinoids: A Novel Approach Using Ultra-High-Performance Supercritical Fluid Chromatography/Photodiode Array-Mass Spectrometry , 2016, Cannabis and cannabinoid research.

[19]  Ethan B. Russo.  Cannabinoids in the management of difficult to treat pain , 2008, Therapeutics and clinical risk management.

[20]  G. Siuzdak,et al.  XCMS Online: a web-based platform to process untargeted metabolomic data. , 2012, Analytical chemistry.

[21]  R. Jubb,et al.  PATIENTS AND METHODS , 1982 .

[22]  Benjamin Debrus,et al.  Innovative development and validation of an HPLC/DAD method for the qualitative and quantitative determination of major cannabinoids in cannabis plant material. , 2009, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[23]  Penny F Whiting,et al.  Cannabinoids for Medical Use: A Systematic Review and Meta-analysis. , 2015, JAMA.

[24]  Thomas A. Briellmann,et al.  Isolation of Δ9-THCA-A from hemp and analytical aspects concerning the determination of Δ9-THC in cannabis products , 2005 .

[25]  S. Burstein,et al.  The cannabinoid acids: nonpsychoactive derivatives with therapeutic potential. , 1999, Pharmacology & therapeutics.

[26]  J. Zhou,et al.  Trigonelline: a plant alkaloid with therapeutic potential for diabetes and central nervous system disease. , 2012, Current medicinal chemistry.

[27]  A. Hanson,et al.  Quaternary Ammonium and Tertiary Sulfonium Compounds in Higher Plants , 1993 .

[28]  M. Zoli,et al.  "Heart-cut" bidimensional achiral-chiral liquid chromatography applied to the evaluation of stereoselective metabolism, in vivo biological activity and brain response to chiral drug candidates targeting the central nervous system. , 2016, Journal of chromatography. A.

[29]  J. McPartland,et al.  Cannabis and Cannabis Extracts: Greater Than the Sum of Their Parts? , 2001 .

[30]  M. Baraldi,et al.  Quantitative analysis of acetylcholine in rat brain microdialysates by liquid chromatography coupled with electrospray ionization tandem mass spectrometry , 2010, Journal of Neuroscience Methods.

[31]  Robert Verpoorte,et al.  Metabolomic differentiation of Cannabis sativa cultivars using 1H NMR spectroscopy and principal component analysis. , 2004, Journal of natural products.

[32]  J. McPartland,et al.  Cannabis is more than simply Δ9-tetrahydrocannabinol , 2003, Psychopharmacology.

[33]  R. Dixon,et al.  Plant metabolomics: large-scale phytochemistry in the functional genomics era. , 2003, Phytochemistry.

[34]  M. Zoli,et al.  7-Chloro-5-(furan-3-yl)-3-methyl-4H-benzo[e][1,2,4]thiadiazine 1,1-Dioxide as Positive Allosteric Modulator of α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid (AMPA) Receptor. The End of the Unsaturated-Inactive Paradigm? , 2016, ACS chemical neuroscience.

[35]  I. Matias,et al.  Antitumor Activity of Plant Cannabinoids with Emphasis on the Effect of Cannabidiol on Human Breast Carcinoma , 2006, Journal of Pharmacology and Experimental Therapeutics.

[36]  M. Zoli,et al.  An improved LC-S/MS method for the quantitation of adenosine concentration in mice brain microdialysates. , 2012, Journal of pharmaceutical and biomedical analysis.

[37]  Simona Pichini,et al.  Evaluation of cannabinoids concentration and stability in standardized preparations of cannabis tea and cannabis oil by ultra-high performance liquid chromatography tandem mass spectrometry , 2017, Clinical chemistry and laboratory medicine.

[38]  K. Müller-Vahl,et al.  The therapeutic potential of cannabis and cannabinoids. , 2012, Deutsches Arzteblatt international.

[39]  David S. Wishart,et al.  MetaboAnalyst 3.0—making metabolomics more meaningful , 2015, Nucleic Acids Res..

[40]  G. Puia,et al.  Design, stereoselective synthesis, configurational stability and biological activity of 7-chloro-9-(furan-3-yl)-2,3,3a,4-tetrahydro-1H-benzo[e]pyrrolo[2,1-c][1,2,4]thiadiazine 5,5-dioxide. , 2014, Bioorganic & medicinal chemistry.

[41]  Jean-Luc Wolfender,et al.  Study of Leaf Metabolome Modifications Induced by UV-C Radiations in Representative Vitis, Cissus and Cannabis Species by LC-MS Based Metabolomics and Antioxidant Assays , 2014, Molecules.

[42]  Maximilian Peters,et al.  HU-331: a cannabinoid quinone, with uncommon cytotoxic properties and low toxicity , 2007, Expert opinion on investigational drugs.

[43]  M. Luginbühl,et al.  Isolation of Delta9-THCA-A from hemp and analytical aspects concerning the determination of Delta9-THC in cannabis products. , 2005, Forensic science international.

[44]  M. Schmid,et al.  Determination of the relative percentage distribution of THCA and Δ(9)-THC in herbal cannabis seized in Austria - Impact of different storage temperatures on stability. , 2015, Forensic science international.

[45]  R. Verpoorte,et al.  Chromatographic and Spectroscopic Data of Cannabinoids from Cannabis sativa L. , 2005 .

[46]  G. Cannazza,et al.  Development of an in vitro liquid chromatography-mass spectrometry method to evaluate stereo and chemical stability of new drug candidates employing immobilized artificial membrane column. , 2014, Journal of chromatography. A.

[47]  M. Huestis,et al.  Blood cannabinoids. I. Absorption of THC and formation of 11-OH-THC and THCCOOH during and after smoking marijuana. , 1992, Journal of analytical toxicology.