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Universidade Federal Do Rio Grande Do Sul Faculdade UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL FACULDADE DE AGRONOMIA PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIA DO SOLO MÉTODOS APLICADOS À RASTREABILIDADE DE Cannabis sativa L. (MACONHA) EM TERRITÓRIO BRASILEIRO Mauro Sander Fett (Tese) UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL FACULDADE DE AGRONOMIA PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIA DO SOLO MÉTODOS APLICADOS À RASTREABILIDADE DE Cannabis sativa L. (MACONHA) EM TERRITÓRIO BRASILEIRO MAURO SANDER FETT Engenheiro Agrônomo (UFRGS) Mestre em Economia Rural (UFRGS) Tese apresentada como um dos requisitos à obtenção do Grau de Doutor em Ciência do Solo Orientador: Prof. Dr. Flávio Anastácio de Oliveira Camargo Porto Alegre (RS) Outubro de 2017 CIP - Catalogação na Publicação Fett, Mauro Sander MÉTODOS APLICADOS A RASTREABILIDADE DE Cannabis sat iva L . (MACONHA ) EM TERRITÓRI O BRASILEIRO I Mauro Sander Fett. -- 2017. 97 f. Ori entador : Flávi o Anastácio de Oli veira Camargo . Tese (Do utorado) -- Universidade Federal do Rio Grande do Sul, Facul dade de Agronomia, Programa de Pós- Graduação em Ciência do Solo, Porto Alegre, BR- RS, 2017 . 1 . Rastreabilidade. 2 . Cannabi s sativa L . 3. Ci ência forense . 4 . Composição química. 5 . Marcadores genéticos . I. Camargo, Flávio Anastácio de Oliveira, orient . II . Título . Elaborada pelo Sistema de Geração Automática de Ficha Catalogrâfica da UFRGS com os dados fornecidos pelo(a) autor(a ). Dedico à minha família, especialmente meu pai e minha mãe. ii AGRADECIMENTOS Ao professor Flávio Camargo pela orientação e amizade no período do doutorado. Aos colegas de trabalho Rafael Ortiz e Eduardo Ávila pelo incentivo e contribuições no trabalho. Aos pesquisadores, amigos e colaboradores deste projeto de estudo, Gleidson Gimenes, Roberta Mariot e Leonardo Capeleto pelo apoio nos trabalhos. Aos professores Tales Tiecher e Valdir Stefenon pelo auxílio no processamento dos dados. À UFRGS e à PUC/RS pelo apoio institucional e estrutura física para realização dos estudos. À Polícia Federal e aos demais colegas de trabalho que apoiaram e colaboraram para que o trabalho pudesse ser realizado. Aos amigos, próximos ou distantes, que colaboraram em manter o andamento de vida “normal” durante este período de estudos. Aos meus pais, Pedro e Janice, à minha esposa, Andrea, e aos meus filhos, Laura e Mateus, pelo apoio e compreensão neste período de estudos e privações. iii METODOS APLICADOS À RASTREABILIDADE DE Cannabis sativa L. (MACONHA) EM TERRITÓRIO BRASILEIRO1 Autor: Mauro Sander Fett Orientador: Prof. Flávio Anastácio de Oliveira Camargo RESUMO A repressão ao tráfico de drogas tem sido um dos principais focos de ação da segurança pública nacional e internacional, devido aos graves problemas de saúde pública associados ao consumo, bem como a grande quantidade de crimes associados ou resultantes deste consumo. Os resultados do Relatório Mundial de Drogas de 2013 das Nações Unidas mostram que a demanda estimada por drogas não está reduzindo consideravelmente. Conforme este relatório, a maconha continua sendo a substância ilícita mais utilizada no mundo, embora tenha havido uma redução no crescimento de seus usuários. Uma das formas de combater o tráfico de drogas, direcionando as investigações e apontando as possíveis rotas, é a identificação de sua origem pela rastreabilidade. Isso pode ser realizado através da observação da composição química ou orgânica do material, baseando‐se no conceito de marcadores. No caso da maconha, em que o uso direto de partes da planta, os marcadores podem ser identificados na composição da própria planta, influenciados pelas características genotípicas das plantas e pelas condições ambientais a que estas estão submedidas durante o desenvolvimento. Assim, foram realizadas análises da concentração química elementar, das relações isotópicas de 15N/14N e 13C/12C e de marcadores genéticos específicos em amostras de Cannabis coletadas em diferentes pontos ou locais de cultivo no Nordeste brasileiro, cultivadas em ambiente controlado e apreendidas em ações policiais, a fim de identificar corretamente o local de cultivo (rastrear sua origem). Os resultados obtidos foram processados no método estatístico multivariado de análise discriminante múltipla. As análises de concentração química elementar e o sistema de marcadores genéticos utilizados para rastrear a origem da Cannabis mostraram capacidade de classificar corretamente as amostras avaliadas, de acordo com o seu local de cultivo ou coleta. Por outro lado, a relação isotópica não foi adequada para separar as amostras de tecido vegetal de Cannabis, de acordo com seu local de cultivo ou coleta no Nordeste brasileiro. Dessa forma, verificou-se que as análises da concentração química elementar e de genotipagem por sistema de marcadores (multiplex 13-loci STR) podem ser utilizadas como marcadores ou indicadores para rastrear a origem de amostras de Cannabis, inclusive com a exatidão necessária para uso policial e forense. _______________________________________________ 1Tese de Doutorado em Ciência do Solo. Programa de Pós-Graduação em Ciência do Solo, Faculdade de Agronomia, Universidade Federal do Rio Grande do Sul. Porto Alegre.(82p.) Outubro, 2017. iv APPLIED METHODS FOR THE TRACEABILITY OF Cannabis sativa L. (MARIJUANA) IN BRAZILIAN TERRITORY1 Author: Mauro Sander Fett Adviser: Prof. Flávio Anastácio de Oliveira Camargo ABSTRACT The repression of drug trafficking has been one of the main action focus of national and international public security, due to the serious public health problems associated to the drug consumption, as well as the large amount of crimes associated or resulting from drug consumption. The results of World Report on Drugs of 2013 by the United Nations show that the estimated demand for drugs is not reducing considerably. According to this report, marijuana continues to be the most used illicit substance in the world, although there had been a reduction in the increase of its users. One of the ways to combat drug trafficking, leading investigations and pointing out possible routes, is the identification of its origin through traceability. That can be accomplished through the observation of the chemical or organic material composition, based on markers concept. In the case of marijuana, in which direct use of parts of the plant, the markers can be identified in the composition of the plant itself, influenced by the plant genotypic characteristics and the environmental conditions to which they are sub- measured during development. Thus, analysis of elemental chemical concentration, stable isotopes analysis (15N/14N and 13C/12C) and specific genetic markers were performed on Cannabis samples collected in different cultivation sites in Brazilian Northeast, samples from plants grown in a controlled environment and police seized samples, in order to identify correctly the cultivation place (trace its origin). The achieved results were processed in the multiple discriminant analysis (MDA). The analysis of elemental chemical concentration and the genetic markers system used to trace the origin of Cannabis showed the ability to classify correctly the evaluated samples, according to their cultivation or collection site. On the other hand, the stable isotopes analysis was not suitable for sorting out the samples of Cannabis plant tissue, in accordance with their cultivation place in Brazilian Northeast. Therefore, it has verified that analysis of elemental chemical concentration and genotyping by markers system (multiplex 13 – loci STR) may be used as markers or indicators to trace out the origin of Cannabis samples, even with the required accuracy for police and forensic use. ______________________________ 1Doctoral thesis in Soil Science – Programa de Pós-Graduação em Ciência do Solo, Faculdade de Agronomia, Universidade Federal do Rio Grande do Sul. Porto Alegre (82p) October, 2017. v SUMÁRIO Pagina 1 INTRODUÇÃO GERAL .................................................................................................... 1 2 REVISÃO BIBLIOGRÁFICA ............................................................................................ 6 2.1 Ciências forenses e criminalística ...................................................................... 6 2.2 Cannabis sativa L. (Maconha) .............................................................................. 9 2.3 Rastreabilidade ....................................................................................................... 16 2.4 Metodologias utilizadas para a rastreabilidade ..............................................19 3 IDENTIFICAÇÃO DA ORIGEM DE Cannabis sativa L. (MACONHA) PELA CONCENTRAÇÃO QUÍMICA ELEMENTAR.................................................................. 24 3.1. Introdução .................................................................................................................... 24 3.2. Material e métodos ..................................................................................................... 25 3.3. Resultados e discussão ........................................................................................... 30 3.2.1. Amostras do Nordeste brasileiro........................................................................ 30 3.3.2. Amostras do Nordeste versus Amostras cultivadas em vaso ................... 43 3.4. Conclusões .................................................................................................................. 46 4 CAPITULO 3. USO DAS RELAÇÕES ISOTÓPICAS DE 15N/14N E 13C/12C PARA RASTREAR O LOCAL DE CULTIVO DE PLANTAS DE MACONHA
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