Industrial hemp is legally defined in the United States by the Agriculture Improvement Act of 2018 (2018 Farm Bill) as Cannabis containing <0.3% total tetrahydrocannabinol (THC). The 2018 Farm Bill does not, however, specify standard methods for sample preparation or quantification of cannabinoids (including THC) in Cannabis. Extraction efficiency of phytochemicals is well-known to depend on the solvent and extraction method used. In this project, we evaluated the effect of sample preparation extraction methods on the quantitative analysis of five cannabinoids found in industrial hemp with regulatory or commercial significance: cannabidiol (CBD), cannabidiolic acid (CBDA), delta-9-tetrahydrocannabinol (THC), tetrahydrocannabinolic acid (THCA), and cannabinol (CBN). Extraction methods evaluated include: QuEChERS, diethyl ether, ethanol, and methanol. Extracts obtained via these methods were subject to quantitative cannabinoid analysis by UPLC/PDA. Standard curves for quantification of each cannabinoid were constructed using authentic standards for quantification. The concentrations of each cannabinoid in the plant material determined via each of the extraction methods were compared using one-way ANOVA followed by Tukey's HSD (significant difference defined as p <0.05). All extraction methods evaluated returned different concentrations of total THC in the plant material. The QuEChERS extraction resulted in the highest calculated concentrations of THC, THCA and CBDA, reporting three to four times greater than obtained via other extractions evaluated. Classification of the starting plant material as hemp or marijuana depended on the extraction method used. These findings clearly and quantitatively demonstrate the need for standardization of extraction methods for hemp analysis and regulation. / Master of Science in Life Sciences / Hemp is a type of Cannabis plant that produces an insignificant amount of the psychoactive cannabinoid tetrahydrocannabinol (THC). Hemp was federally illegal in the United States until the 2018 Farm Bill differentiated hemp from Marijuana, defining hemp as containing less than 0.3% total THC. Standard methods for cannabinoid testing in hemp have not been defined. In this project, four extraction methods with potential use for sample preparation in hemp analysis were evaluated and compared. The extraction methods evaluated included: ethanol, methanol, QuEChERS, and diethyl ether. The concentrations of cannabinoids in each of the plant extracts were then determined using an appropriate analytical method and authentic standards for Cannabidiol (CBD), Cannabidiolic Acid (CBDA), THC, Tetrahydrocannabinolic Acid (THCA), and Cannabinol (CBN). Total concentrations of each cannabinoid in the plant material were then calculated using each extraction method. All extraction methods evaluated resulted in different concentrations of total THC in the plant material, with QuEChERS resulting in the highest calculated concentrations of THC, THCA, and CBDA. The identify of this plant material as hemp or marijuana depended on the extraction method used. This result is not surprising, as extraction efficiency of phytochemicals is well known to depend on the solvent and extraction method used. Nonetheless, our findings clearly demonstrate the need for standardization of extraction methods for hemp analysis and regulation.

A Cannabis sativa L. é a droga de abuso de uso proscrito mais consumida no mundo. O consumo desta droga por condutores de veículo automotor está associado com o aumento do risco de acidentes de trânsito, com o aumento da gravidade e com o aumento das taxas de mortalidade. O objetivo deste trabalho foi propor métodos analíticos aplicáveis à rotina laboratorial forense para monitoramento do consumo de canábis por condutores. Para tanto, foi utilizada como metodologia a extração líquido-líquido seguida de análise por cromatografia em fase gasosa acoplada a detector de massas para pesquisa de canabinoides em matrizes biológicas como sangue total (ST) e fluido oral (FO). A análise de 11-nor-9-carbóxi-delta9-THC (THC-COOH) em amostras de ST por cromatografia gasosa envolve a derivatização desta molécula e, consequentemente, do THC extraído desta matriz (quando presente), uma vez que ambos estão presentes no mesmo extrato, constituindo mais uma etapa analítica e que requer maior controle das condições de reação. Por outro lado, a detecção de THC em FO pode ser realizada sem a realização desta etapa, o que constitui uma vantagem analítica. A determinação de THC-COOH e THC em ST não demonstrou repetibilidade, o que inviabilizou as análises qualitativas e quantitativas nesta matriz. A detecção de THC em FO se mostrou uma análise simples e passível de validação, porém com limite de detecção (200ng/mL) acima do recomendado pelas guias forenses internacionais (2 ng/mL). A metodologia analítica desenvolvida se mostrou compatível com aplicação na análise confirmatória em casos de intoxicação aguda pelo consumo de canábis, demonstrando a necessidade de utilização de técnicas de concentração de amostras como extração em fase sólida ou microextração em fase sólida, para obtenção de menores limites de detecção, podendo assim ser aplicado na rotina laboratorial para o monitoramento do uso frequente de canábis, e não apenas em casos de intoxicação aguda. Além disso, foi realizada uma abordagem sobre a interpretação da detecção de THC em diferentes matrizes biológicas. / Cannabis sativa L. is the illegal drug of abuse most consumed in the world. The consumption of this drug by motor vehicle drivers is associated with an increased risk of traffic accidents, increased severity and increased mortality rates. The objective of this work was to propose analytical methods applicable to forensic laboratories to verify the consumption of cannabis by drivers. Liquid-liquid extraction followed by gas chromatography (GC) coupled to mass spectrometry (MS) detector has been applied to cannabinoid analysis in biological samples such as whole blood (WB) and oral fluid (OF). The analysis of 11-nor-9-carboxy-delta9-THC (THC-COOH) in WB by GC required the derivatization of this molecule and also involved the derivatization of THC since both were extracted from the same sample. The derivatization constitutes another analytical step, which requires greater control of the reaction conditions. Fortunately, the detection of only THC in OF can be done without performing this step. The determination of THC-COOH and THC in WB did not demonstrate repeatability, which impaired the qualitative and quantitative analyzes in this matrix. The detection of THC in OF proved to be a simple analysis, that could be validated, but the limit of detection (200ng/mL) was higher than the recommended by the international forensic guides (2 ng/mL). The chromatographic method developed was compatible with the application of a confirmatory analysis in acute cannabis intoxication, demonstrating the need to use techniques of samples concentration such as SPE or SPME, in order to obtain lower limits of detection, thus being able to be applied in the laboratory routine for the monitoring of the frequent use of cannabis, and not only in cases of acute intoxication. In addition, it was made an approach of THC detection in different biological matrices.

A Cannabis sativa L. é a droga de abuso de uso proscrito mais consumida no mundo. O consumo desta droga por condutores de veículo automotor está associado com o aumento do risco de acidentes de trânsito, com o aumento da gravidade e com o aumento das taxas de mortalidade. O objetivo deste trabalho foi propor métodos analíticos aplicáveis à rotina laboratorial forense para monitoramento do consumo de canábis por condutores. Para tanto, foi utilizada como metodologia a extração líquido-líquido seguida de análise por cromatografia em fase gasosa acoplada a detector de massas para pesquisa de canabinoides em matrizes biológicas como sangue total (ST) e fluido oral (FO). A análise de 11-nor-9-carbóxi-delta9-THC (THC-COOH) em amostras de ST por cromatografia gasosa envolve a derivatização desta molécula e, consequentemente, do THC extraído desta matriz (quando presente), uma vez que ambos estão presentes no mesmo extrato, constituindo mais uma etapa analítica e que requer maior controle das condições de reação. Por outro lado, a detecção de THC em FO pode ser realizada sem a realização desta etapa, o que constitui uma vantagem analítica. A determinação de THC-COOH e THC em ST não demonstrou repetibilidade, o que inviabilizou as análises qualitativas e quantitativas nesta matriz. A detecção de THC em FO se mostrou uma análise simples e passível de validação, porém com limite de detecção (200ng/mL) acima do recomendado pelas guias forenses internacionais (2 ng/mL). A metodologia analítica desenvolvida se mostrou compatível com aplicação na análise confirmatória em casos de intoxicação aguda pelo consumo de canábis, demonstrando a necessidade de utilização de técnicas de concentração de amostras como extração em fase sólida ou microextração em fase sólida, para obtenção de menores limites de detecção, podendo assim ser aplicado na rotina laboratorial para o monitoramento do uso frequente de canábis, e não apenas em casos de intoxicação aguda. Além disso, foi realizada uma abordagem sobre a interpretação da detecção de THC em diferentes matrizes biológicas. / Cannabis sativa L. is the illegal drug of abuse most consumed in the world. The consumption of this drug by motor vehicle drivers is associated with an increased risk of traffic accidents, increased severity and increased mortality rates. The objective of this work was to propose analytical methods applicable to forensic laboratories to verify the consumption of cannabis by drivers. Liquid-liquid extraction followed by gas chromatography (GC) coupled to mass spectrometry (MS) detector has been applied to cannabinoid analysis in biological samples such as whole blood (WB) and oral fluid (OF). The analysis of 11-nor-9-carboxy-delta9-THC (THC-COOH) in WB by GC required the derivatization of this molecule and also involved the derivatization of THC since both were extracted from the same sample. The derivatization constitutes another analytical step, which requires greater control of the reaction conditions. Fortunately, the detection of only THC in OF can be done without performing this step. The determination of THC-COOH and THC in WB did not demonstrate repeatability, which impaired the qualitative and quantitative analyzes in this matrix. The detection of THC in OF proved to be a simple analysis, that could be validated, but the limit of detection (200ng/mL) was higher than the recommended by the international forensic guides (2 ng/mL). The chromatographic method developed was compatible with the application of a confirmatory analysis in acute cannabis intoxication, demonstrating the need to use techniques of samples concentration such as SPE or SPME, in order to obtain lower limits of detection, thus being able to be applied in the laboratory routine for the monitoring of the frequent use of cannabis, and not only in cases of acute intoxication. In addition, it was made an approach of THC detection in different biological matrices.

A Cannabis sativa L. é a droga de abuso de uso proscrito mais consumida no mundo. O consumo desta droga por condutores de veículo automotor está associado com o aumento do risco de acidentes de trânsito, com o aumento da gravidade e com o aumento das taxas de mortalidade. O objetivo deste trabalho foi propor métodos analíticos aplicáveis à rotina laboratorial forense para monitoramento do consumo de canábis por condutores. Para tanto, foi utilizada como metodologia a extração líquido-líquido seguida de análise por cromatografia em fase gasosa acoplada a detector de massas para pesquisa de canabinoides em matrizes biológicas como sangue total (ST) e fluido oral (FO). A análise de 11-nor-9-carbóxi-delta9-THC (THC-COOH) em amostras de ST por cromatografia gasosa envolve a derivatização desta molécula e, consequentemente, do THC extraído desta matriz (quando presente), uma vez que ambos estão presentes no mesmo extrato, constituindo mais uma etapa analítica e que requer maior controle das condições de reação. Por outro lado, a detecção de THC em FO pode ser realizada sem a realização desta etapa, o que constitui uma vantagem analítica. A determinação de THC-COOH e THC em ST não demonstrou repetibilidade, o que inviabilizou as análises qualitativas e quantitativas nesta matriz. A detecção de THC em FO se mostrou uma análise simples e passível de validação, porém com limite de detecção (200ng/mL) acima do recomendado pelas guias forenses internacionais (2 ng/mL). A metodologia analítica desenvolvida se mostrou compatível com aplicação na análise confirmatória em casos de intoxicação aguda pelo consumo de canábis, demonstrando a necessidade de utilização de técnicas de concentração de amostras como extração em fase sólida ou microextração em fase sólida, para obtenção de menores limites de detecção, podendo assim ser aplicado na rotina laboratorial para o monitoramento do uso frequente de canábis, e não apenas em casos de intoxicação aguda. Além disso, foi realizada uma abordagem sobre a interpretação da detecção de THC em diferentes matrizes biológicas. / Cannabis sativa L. is the illegal drug of abuse most consumed in the world. The consumption of this drug by motor vehicle drivers is associated with an increased risk of traffic accidents, increased severity and increased mortality rates. The objective of this work was to propose analytical methods applicable to forensic laboratories to verify the consumption of cannabis by drivers. Liquid-liquid extraction followed by gas chromatography (GC) coupled to mass spectrometry (MS) detector has been applied to cannabinoid analysis in biological samples such as whole blood (WB) and oral fluid (OF). The analysis of 11-nor-9-carboxy-delta9-THC (THC-COOH) in WB by GC required the derivatization of this molecule and also involved the derivatization of THC since both were extracted from the same sample. The derivatization constitutes another analytical step, which requires greater control of the reaction conditions. Fortunately, the detection of only THC in OF can be done without performing this step. The determination of THC-COOH and THC in WB did not demonstrate repeatability, which impaired the qualitative and quantitative analyzes in this matrix. The detection of THC in OF proved to be a simple analysis, that could be validated, but the limit of detection (200ng/mL) was higher than the recommended by the international forensic guides (2 ng/mL). The chromatographic method developed was compatible with the application of a confirmatory analysis in acute cannabis intoxication, demonstrating the need to use techniques of samples concentration such as SPE or SPME, in order to obtain lower limits of detection, thus being able to be applied in the laboratory routine for the monitoring of the frequent use of cannabis, and not only in cases of acute intoxication. In addition, it was made an approach of THC detection in different biological matrices.

Recent evidence suggests that sustained consumption of a high-fat diet is associated with reduced CB1 receptor expression in some brain areas. Many of the neuromodulatory functions of endogenous cannabinoids are mediated by the CB1 receptor. The CB1 receptor also mediates the behavioral and physiological effects of delta-9-tetrahydrocannabinol (delta-9-THC), the primary psychoactive constituent of marijuana. While high-fat diets are associated with region-specific changes in CB1 receptor expression, it is not clear whether such changes are behaviorally relevant. To that end, separate groups of male and female rats were placed on either a high-fat diet or a standard diet. Cannabinoid function was determined in a triad of measures (e.g., hypothermia, gross locomotion, time on bar apparatus) at postnatal day 30 (PD30), PD44, PD68 and PD114. These age points respectively correspond to rodent models of early adolescence, late adolescence, early adulthood and full maturity in humans. Male rats were also tested at PD37 and PD61. Subsequently, the antinociceptive properties of delta-9-THC and the effect of delta-9-THC on food intake were also measured. After 38 days, female rats maintained on a high-fat diet were significantly less sensitive to the psychomotor effects of delta-9-THC than were the female rats maintained on the control diet. These diet-related differences persisted into full maturity. Female rats maintained on a high-fat diet were also less sensitive to changes in food intake caused by delta-9-THC than were female rats maintained on the control diet. In contrast, the hypothermic effects of delta-9-THC were not differentially affected by the type of diet consumed. Likewise, female rats maintained on a high-fat diet exhibited tail-flick latencies that were indistinguishable from those of female rats maintained on the control diet. With two minor exceptions, and in sharp contrast to female rats, sensitivity to the pharmacological effects of delta-9-THC was not differentially affected by the type of diet in male rats. In short, female rats maintained on a high-fat diet appeared to be cross-tolerant to the psychomotor and hyperphagic effects of delta-9-THC while male rats maintained on a high-fat diet exhibited responses to delta-9-THC that were virtually indistinguishable from control animals.

En metod som kan användas för att extrahera opolära narkotika- och läkemedelssubstanser ur matriser med ett högt fettinnehåll har tagits fram. Metoden är baserad på ett extraktionsprotokoll från Livsmedelsverket (SLV K1-f4-m018.3) som används för analys av bekämpningsmedel i animaliska livsmedel. Metoden har sedan optimerats för forensiska applikationer genom att ändra olika variabler. Två av variablerna som testades var olika typer av extraktionsmedel och mängden tillsatt C18 (oktadekylsilyl)-sorbent. Experimenten visade att acetonitril som extraktionsmedel och en ökad tillsats av C18 eliminerade den största andelen av fettmolekylerna i matrisen utan att kompromissa med utbytet av den undersökta analyten. Analyterna som undersöktes var 5F-AKB48, tetrahydrocannabinol (THC) och alprazolam och den fettinnehållande matrisen utgjordes av smör. Metoden lyckades minska fetthalten i smörproverna från ca 80- till 3-5 % fett samtidigt som ett utbyte av analyterna erhölls på 70-90 %. Analyten med högst utbyte efter extraktionen var 5F- AKB48 med ett utbyte på cirka 90 %. Utbytet för alprazolam och THC blev 72- respektive 75 %. Vid extraktion med etylacetat, som var det extraktionsmedel som användes i den ursprungliga metoden (SLV K1-f4-m018.3), erhölls inga minskningar av fetthalten överhuvudtaget utan hamnade runt 80 % i samtliga analyser. När metodens kapacitet testades med ökade fetthalter visade det sig att fetthalten efter extraktionen blev lägre i de prover som hade en högre fetthalt från början. Utbytet av analyten (5F-AKB48 i detta fall) påverkades däremot inte nämnvärt av de ökade fetthalterna utan höll sig stabilt runt 85 %.

Cannabinoids suppress Th1 immunity in a variety of models including infection with the intracellular pathogen Legionella pneumophila (Lp). To examine the cellular mechanism of this effect, mouse bone marrow-derived dendritic cells (DCs) were studied following infection and drug treatment. DCs produced high levels of IL-12p40 following Lp infection. THC suppressed this cytokine response in a concentration-dependent manner and the endocannabinoids 2-arachidonoyolglycerol and virodhamine less potently suppressed cytokine production. DCs expressed mRNA for cannabinoid receptor 1 (CB1), CB2, and transient receptor potential vanilloid type 1 (TRPV1); furthermore, inhibition of Gi signaling by adding pertussis toxin completely attenuated the suppression induced by low concentrations of THC but not at high concentrations. In addition, the THC suppression was partially attenuated in DC cultures from CB1 and CB2 knockout mice and in cultures from normal mice co-treated with THC and cannabinoid receptor antagonists. Cytokine suppression was not attenuated by pretreatment with the TRPV1 antagonist capsazepine, suggesting that Gi signaling and cannabinoid receptors, but not TRPV1, are involved in THC-induced suppression of DC potential to polarize the development of naïve T cells to be Th1 cells. Besides IL-12, THC suppressed other DC polarizing characteristics such as the expression of MHC class II and co-stimulatory molecules CD86 and CD40, as well as the Notch ligand Delta 4. However, THC treatment did not affect other DC functions such as intracellular killing of Lp and Lp-induced apoptosis. Testing the capacity of THC to suppress DC polarizing function with T cells showed that DCs infected in vitro with Lp were able to immunize mice when injected prior to a lethal Lp infection; however, the immunization potential along with Th1 cytokine production was attenuated by THC treatment of the cells at the time of in vitro infection. In addition, THC-treated and Lp-infected DCs poorly stimulated primed splenic CD4 T cells in culture to produce IFN-gamma (IFN-y); however, this stimulating deficiency was reversed by adding recombinant IL-12p40 protein to the cultures. In conclusion, the data suggest that THC inhibits Th1 polarization by targeting essential DC functions such as IL-12p40 secretion and the maturation and expression of co-stimulatory and polarizing molecules.

High density electroencephalographic (EEG) measures were used to assess the effects of acute delta 9-tetrahyrdrocannabidol (THC) administration on extinction of conditioned fear responses. Fear conditioning was initiated using a differential classical conditioning paradigm that paired an aversive unconditioned stimulus (shock) with a signaling stimulus (CS+), whereas another stimulus served as a safety signal (CS-). Evoked potentials, induced event-related spectral perturbations (ERSP), and associated intertrial coherence (ITC) measures were used to quantify the acquisition and extinction of conditioned fear responses. Participants (N = 10 males) exhibited conditioning to the CS+ across fear acquisition training, as reflected by greater late positive (posterior sites) and late negative (anterior sites) potential amplitude to the CS+ relative to the CS-. Acute administration of THC facilitated extinction of the conditioned response to the CS+ relative to placebo, as reflected by greater LPP and LNP amplitude to the CS+ relative to the CS- in the placebo, but not THC condition. ERSP analyses suggest the lack of difference between CS+ and CS- ERP amplitude may be partially explained by a shifting of attention from external stimuli to internal processing in the THC condition. However, relative to placebo, THC administration also increased the amplitude of some measures of the conditioned response (LNP) to the CS-, suggesting a generalization of fear or lack of discrimination in this condition.

Submitted by Maykon Nascimento (maykon.albani@hotmail.com) on 2014-12-19T17:57:04Z No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertacao Identificação Química em Nível Molecular de Amostras de de Maconha por ESI-FT-ICR MS.pdf: 5116408 bytes, checksum: bd1abd84e579ae77841f93ccc152cb2f (MD5) / Approved for entry into archive by Elizabete Silva (elizabete.silva@ufes.br) on 2014-12-19T18:13:41Z (GMT) No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertacao Identificação Química em Nível Molecular de Amostras de de Maconha por ESI-FT-ICR MS.pdf: 5116408 bytes, checksum: bd1abd84e579ae77841f93ccc152cb2f (MD5) / Made available in DSpace on 2014-12-19T18:13:41Z (GMT). No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertacao Identificação Química em Nível Molecular de Amostras de de Maconha por ESI-FT-ICR MS.pdf: 5116408 bytes, checksum: bd1abd84e579ae77841f93ccc152cb2f (MD5) Previous issue date: 2014 / mais consumida no país, e proscrita pela Lei n° 11.343 de 23 de agosto de 2006 (chamada de “nova lei de droga”), onde todos os isômeros, sais, éteres e ésteres do ∆9-Tetrahidrocannabinol (THC), princípio ativo, foram proscritos. O método utilizado pela Polícia Civil do Estado do Espírito Santo para a identificação de cannabinóides é o teste colorimétrico, por meio de solução básica de Salt Fast Blue B, o qual apresenta resultados falsos negativos e falsos positivos. A técnica de espectrometria de massas de altíssima resolução e exatidão de massas (ESI(-)FTICR MS), permite detectar os principais cannabinóides na forma de molécula desprotonada, íon [M-H]-. Alguns íons que podem ser identificados são: [CBN - H]- de m/z 309 (CBN = cannabinol); [THC - H]- de m/z 313 (THC = tetrahidrocannabinol) e [CBD - H]- de m/z 313; [CBC - H]- de m/z 327 (CBC = cannabicromeno); [CBEA - H]- de m/z 345 (CBEA = ácido cannabielsóico); [CBNA - H]- de m/z 353 (CBNA = ácido cannabinólico); [THCA - H]- de m/z 357 (THCA = ácido tetrahidrocannabinólico); [8α, 11-Bis-hydroxy-∆9-THC-A - H]- de m/z 389); [∆9-THCA +C2H2O - H]- de m/z 357; e dímeros com m/z de 637, 653, 673, 681, 685 e 717. Foram encontrados adulterantes identificados como [M + N + H]+ : 491; [2M + N + H]+ : 819 e [3M + N + H]+ : 1147, onde M = OTHC (328Da C21H28O3) e N = Nicotina (162Da C10H14N2), além de lidocaína e cocaína. Ainda foram identificados alguns noncannabinóides como Cannflavino A e B e ácidos graxos como palmítico, oleico, linolênico e gama-linolênico nos extratos de sementes de Cannabis. Este estudo tem o objetivo de identificar o perfil químico de amostras de maconha, apreendidas pela Polícia Civil do Estado do Espírito Santo, por ESI(±)-FT-ICR MS. / The Cannabis sativa L. is well known in Brazil as "maconha". This is the most consumed drug in this country, proscribed by the Law number 11.343 of 23rd August 2006(called "new drug law) , where all isomers, salts, ethers and esters of ∆9Tetrahidrocannabinol (THC), active principle, were proscribed. The method used by the Civil Police of Espírito Santo state to identify the cannabinoids is the test called "colorimetric. It is used by a basic solution of Salt Fast Blue B, which presents results false negatives and false positive. The technic of mass spectrometry of high solution and mass accuracies, ESI(-)FT-ICR MS, allows to detect the main cannabinoids in the form of molecules deprotonated , ions [M-H]. Some ions that can be identified are: [CBN-H]- of m/z 309 (CBN = cannabinol); [THC - H]- of m/z 313 (THC = tetrahidrocannabinol) and [CBD - H]- of m/z 313; [CBC –H]- of 327 (CBC = cannabicromeno); [CBEA - H]- of m/z 345 (CBEA = acid cannabielsóico); [CBNA - H] of m/z 353 (CBEA = acid cannabinólico); [THCA-H] de m/z 357 (THCA = acid tetrahidrocannabinólico); [8α,11-Bis-hydroxy-∆9-THC-A - H]- de m/z 389); [∆9-THCA +C2H2O - H]- of m/z 357; and dimers with m/z 637, 653, 673, 681, 685 and 717. Identified as contaminants found: [M + H]+: 491; [2M + N + H]+: 819; [3M + N + H]+: 1147, where M = OTHC (328Da C21H28O3) and N is nicotine (C10H14N2 162Da) beyond lidocaine and cocaine. Still some noncannabinóides were identified as: Cannflavino A and B and fatty acids such as palmitic, oleic, linoleic and gammalinolenic acid in the extracts of cannabis seeds. This study has the purpose to identify the chemical profile from samples of cannabis seized by the Civil Police from Espirito Santo state, by ESI(±)-FT-ICR MS.

The frontostriatal dopamine system modulates brain function and is affected by both genetic and environmental factors. Dysfunction of this system is associated with many pathological states, including schizophrenia. The enzyme catechol-O- methyltransferase (COMT) metabolises dopamine and its gene contains a polymorphism (Val¹⁵⁸Met) that affects enzyme activity. Delta-9- tetrahydrocannabinol (THC), the main psychoactive component of cannabis, has been suggested to interact with this polymorphism to increase the risk for psychosis and cognitive impairments. Dopaminergic mechanisms are a plausible candidate for mediating this interaction. I used microdialysis coupled with high performance liquid chromatography (HPLC) to examine the effects of THC on extracellular dopamine and its metabolites in the nucleus accumbens, dorsal striatum and medial prefrontal cortex (mPFC) in freely moving mice. Following acute COMT inhibition with tolcapone, THC increased extracellular dopamine levels in the nucleus accumbens in tolcapone-, but not in vehicle-, treated mice. The introduction of the low activity Met allele into the COMT gene produced a highly specific, novel mouse model of the Val158Met polymorphism. In contrast to the effects of acute COMT inhibition, the Met allele protected against THC-induced changes in accumbal dopamine. No interactive neurochemical effects were observed in the dorsal striatum (pharmacological and genetic study) or in a preliminary study of the mPFC (genetic study only). On a progressive ratio task measuring motivational salience, the direction of the interactive effect between COMT genotype and THC differed between 2 independent cohorts and provided tentative leads that stress/arousal-dependent effects on COMT may have a confounding effect. My data provide evidence that COMT activity modulates the effect of THC on accumbal dopamine function, and suggest the mechanism through which this interaction is mediated differs between acute and lifelong reduction in COMT activity. Through the interactive effect on the dopaminergic system, the data provide a potential mechanism for the reported interaction between COMT and cannabis/THC in determining psychosis risk and cognitive impairments.