Determinação de etanol em fluido oral por headspace associada à cromatografia gasosa com detector de ionização em chama

Orientadora : Professora Dra. Renata Pereira LimbergerMonografia (especialização) - Universidade Federal do Paraná, Setor de Ciências da Saúde, Curso de Especialização em Análises ClínicasInclui referênciasResumo: O etanol é o principal constituinte de bebidas alcoólicas. Quando consumido de forma abusiva é a principal causa para a ocorrência de acidentes de trânsito. Um método por cromatografia gasosa com detector de headspace de ionização em chama para determinação de etanol em fluido oral foi validado. A coleta de fluido oral foi realizada utilizando um dispositivo QuantisalTM e a separação foi realizada em uma coluna de ZB-BAC1 (30mx0,32mmx1,80µm). O método apresentou especificidade adequada, linearidade >0,997, precisão (0,86-11,91%) e exatidão (92- 107%) aceitáveis. Limite de detecção 0,0019 g/L e limite de quantificação 0,005 g/L de etanol em amostras de fluido oral

Determinação de etanol em trabalhadores de postos de gasolina

Milhões de pessoas morrem anualmente vítimas de acidentes de trânsito, sendo grande parte dos acidentes correlacionados ao ato de dirigir sob influência (DUI) de álcool ou drogas. No Brasil, os trabalhadores de postos de gasolina ficam expostos diariamente, durante a jornada de trabalho, ao etanol, presente no etanol combustível e utilizado como aditivo na gasolina. O objetivo deste trabalho foi avaliar a possível interferência do etanol presente nos combustíveis aos quais os frentistas estão expostos diariamente durante a jornada de trabalho, no teste do etilômetro e analisar amostras de fluido oral destes trabalhadores por headspace acoplado a cromatógrafo a gás com detector de massas (HS-CG/EM). Foram convidados a participar do estudo frentistas de 26 postos de gasolina. Primeiramente foi aplicado um questionário, na sequência realizado o teste do etilômetro e coletada amostra de fluido oral através do dispositivo de coleta QuantisalTM para posterior análise em laboratório. Todas as amostras de fluido oral foram analisadas segundo método validado. Em 100% das amostras de fluido oral foi observado presença de etanol e destas, em 72,83% havia presença de etanol em níveis acima do limite de quantificação do método. Com relação ao teste do etilômetro, apenas uma amostra de ar exalado (0,62%) apresentou resultado positivo. As diferenças de concentração de etanol nas diferentes matrizes biológicas são explicadas pelo fato da concentração de etanol ser dependente da concentração de água presente nas matrizes. Os resultados positivos nas amostras de FO e os resultados negativos nas amostras de ar exalado podem ser explicados pela possibilidade do método de análise em FO por HS-CG/EM ser mais sensível do que o teste do etilômetro.Millions of people die from traffic accidents, and a large proportion of accidents related to driving under the influence (DUI) of alcohol or drugs. In Brazil, gasoline station attendants are exposed daily during the workday, to ethanol in the fuel and ethanol used as an additive in gasoline. The aim of this study was to evaluate the possible interference of ethanol present in the fuels ethanol and gasoline, to which the attendants are exposed, through breathalyzer and oral fluid (OF) analysis by headspace gas chromatography mass spectrometry (HS-GC/MS). Gasoline stations attendants of 26 gasoline stations were invited to participate in the study. First a questionnaire was fulfilled, followed by the breathalyzer and the OF collection with Quantisal® device for laboratory analysis. All OF samples were analyzed by HS-GC/MS using a validated method. The presence of ethanol was found in 100% of which 72.83% had concentrations above the quantification limit of the method. Regarding the breathalyzer, only one expired air sample (0.62%) had a positive result. The differences in ethanol concentration in different biological matrices are explained by the fact that ethanol concentration is dependent on the concentration of water present in the biological matrix. The positive results in OF samples and negative results in exhaled air samples can be explained by the possibility of the OF analysis by HS-GC/MS to be more sensitive than the breathalyzer

CANNABIS SATIVA: A SYSTEMATIC REVIEW OF PLANT ANALYSIS

Background: Cannabis has been the most widely used illicit drug worldwide throughout many years. Reports from different countries indicate that the potency of cannabis preparation has been increasing, as well as the ratio of tetrahydrocannabinol/cannabidiol has been changing. The high consumption along with the changing chemical profile of the drug has led increasingly to the interest in researching the cannabis plant. Methods: This article reviews available literature on the analytical methods currently used for the detection and quantification of cannabinoids in cannabis plant. The papers were screened by two researchers independently and following a pre-specified protocol. Results and Discussion: The systematic review of the literature allowed to include 42 citations on cannabis plant analysis. Conclusions: The analytical methods for cannabis material published in the included articles of this systematic review showed a lack of relevant information of the development of methods on GC and LC analysis and the limits of detection and quantification of mass detectors

Multianalytical Method Validation for Qualitative and Quantitative Analysis of Solvents of Abuse in Oral Fluid by HS-GC/MS

The use of oral fluid as a biological matrix to monitor the use of drugs of abuse is a global trend because it presents several advantages and good correlation to the blood level. Thus, the present work aimed to develop and validate an analytical method for quantification and detection of solvents used as inhalants of abuse in oral fluid (OF), using Quantisal™ as collector device by headspace and gas chromatography coupled with a mass detector (HS-GC/MS). Chromatographic separation was performed with a ZB-BAC1 column and the total time of analysis was 11.8 min. The method showed good linearity (correlation coefficient higher than 0.99 for all solvents). The limits of detection ranged from 0.05 to 5 mg/L, while the lower limits of quantification ranged from 2.5 to 12.5 mg/L. Accuracy, precision, matrix effect, and residual effect presented satisfactory results, meeting the criteria accepted for the validation of bioanalytical methods. The method showed good selectivity considering that, for solvents coeluting at the same retention time, resolution was performed by the mass detector. The method developed proved to be adequate when applied in OF samples from users of drugs and may be used to monitor the abuse of inhalants in routine forensic analyses

Caracterização química da planta Cannabis sativa L. a partir de sementes apreendidas pela Polícia Federal no Estado do Rio Grande do Sul

A Cannabis sativa L. (canábis) contém centenas de substâncias químicas em diferentes classes, contudo, a classe dos canabinoides é encontrada unicamente nesta espécie. Popularmente conhecida como maconha, a canábis é a droga ilícita mais consumida no mundo, sendo o Δ9-tetrahidrocanabinol (Δ9-THC), o principal canabinoide de interesse toxicológico, por ser o responsável pela maioria dos efeitos psicotomiméticos ocasionados e estar associado ao uso abusivo da planta. De outro modo, há alguns anos, vem crescendo significativamente o interesse em pesquisas científicas com a canábis, devido aos importantes efeitos terapêuticos que alguns canabinoides têm apresentado. O canabidiol (CBD), por exemplo, é um canabinoide que além de modular os efeitos eufóricos do Δ9-THC, tem apresentado importantes atividades farmacológicas. Embora haja um crescente aumento nas toneladas de apreensões de canábis realizadas no Brasil, não existem dados que caracterizem o perfil químico e a potência da droga que é utilizada ilegalmente no país. Outro aspecto singular com relação às apreensões de canábis refere-se à nova forma de tráfico internacional de drogas, que vem crescendo significativamente no país. Trata-se da remessa de sementes de canábis em pequenas quantidades através de empresas de transporte. Desta forma, este estudo foi realizado a partir de sementes de canábis apreendidas pela Superintendência de Polícia Federal no Rio Grande do Sul. As sementes foram germinadas e as plântulas foram cultivadas, secas e analizadas, sendo todos os passos em condições controladas. As metodologias analíticas utilizadas foram: espectroscopia no infravermelho próximo (NIRS), espectrometria de massas por ressonância ciclotrônica de íons com transformada de Fourier (FT-ICR MS) e cromatografia à gás com detector de massas (CG/EM). O elevado número de compostos presente nas plantas torna os dados obtidos muito complexos, onde pequenas diferenças entre amostras podem ser negligenciadas. Por isso, foram utilizadas as seguintes ferramentas quimiométricas: HCA, PCA, PLS-DA e SVM-DA. As diferentes metodologias de análise utilizadas neste trabalho, permitiram a análise de um total de 73 amostras de canábis que forneceram dados sob químicos, bem como aspectos distintos e relevantes em toxicologia. As informações obtidas por NIRS associadas às ferramentas quimiométricas HCA e PCA possibilitaram agrupar 29 amostras de canábis em diferentes períodos de crescimento; e as ferramentas quimiométricas PLS-DA e SVM-DA permitiram classificar 29 amostras de canábis em diferentes períodos de crescimento com 100% de acerto. O altíssimo poder de resolução da FT-ICR MS possibilitou a identificação 123 compostos canabinoides e metabólitos a partir das análises realizadas em 73 amostras de canábis, o que resultou na compilação de um abrangente perfil químico de canabinoides, até então não obtido. A eficiência de separação da CG acoplada à identificação dos compostos utilizando EM tornou possível a diferenciação de 73 amostras de canábis em tipo fibra ou tipo droga, além da caracterização do perfil químico de canabinoides e terpenoides presentes nas amostras, obtido pela análise em CG/EM. Desta forma, os dados gerados com o desenvolvimento desta tese possibilitaram a identificação e o conhecimento do perfil químico de amostras de canábis apreendidas no Brasil, apresentando informações até então desconhecidas pela polícia brasileira, bem como pelos profissionais da saúde, comunidade científica, órgãos governamentais e políticos do país e também para a população em geral.Cannabis sativa L. (cannabis) contains hundreds of different classes of chemicals, however, the class of cannabinoids is found only in this plant. Popularly known marijuana, cannabis is the most used illicit drug worldwide, and the Δ9-tetrahydrocannabinol (Δ9-THC), the main cannabinoid regarding toxicological interest, being responsible by the majority of psychotomimetic effects and it is associated with the abuse of the plant. Otherwise, for a few years has grown significantly the interest and scientific research with cannabis due to significant therapeutic effects that certain cannabinoids have shown. The cannabidiol (CBD), for example, is a cannabinoid which in addition to modulate the euphoric effects of Δ9-THC, it has shown significant pharmacological activity. Although there is a growing increase in tons of cannabis seizures conducted in Brazil, there are no data which characterize the chemical profile and potency of the drug that is illegally used in the country. Another unique aspect regarding cannabis seizures refers to the new form of international drug trafficking that has grown significantly in the country. This is the shipment of cannabis seeds in small quantities by transportation companies. Thus, this study was based on samples from cannabis seeds seized by the Superintendence of the Federal Police in Rio Grande do Sul. The seeds were germinated and the seedlings were grown, dried and analyzed, all of the steps under controlled conditions. The analytical methods used were: near infrared spectroscopy (NIRS), Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) and gas chromatography coupled with mass spectrometry (GC/MS). The high number of compounds presente in plants renders them very complex data set, where small differences between samples may be wasted. Therefore, the following chemometric tools were used: HCA, PCA, PLS-DA and DA-SVM. The different methods of analysis used in this study allowed the analysis of a total of 73 samples of cannabis that provided data on chemical and distinct and relevant aspects in toxicology. Information obtained by NIRS chemometric tools associated with HCA and PCA group allowed 29 samples of cannabis in different periods of growth; and chemometric tools PLS-DA and SVM-DA can be classified 29 samples of cannabis in different periods of growth with 100% accuracy. The very high resolution power of FT-ICR MS possible to identify 123 compounds cannabinoids and metabolites from the analyzes carried out on 73 samples of cannabis, which resulted in the compilation of a comprehensive chemical profile of cannabinoids, has not been surpassed. The GC separation efficiency coupled with the identification of compounds using MS made it possible to differentiate 73 cannabis samples in type or fiber type drug, besides the characterization of the chemical profile of cannabinoids and terpenoids present in the samples obtained through the analysis on GC/MS. Thus, the data generated with the development of this thesis allowed the identification and knowledge of the chemical profile of cannabis samples seized in Brazil, presenting information previously unknown to Brazilian police as well as by health professionals, the scientific community, government agencies and politicians of the country and also to the general population

Caracterização química da planta Cannabis sativa L. a partir de sementes apreendidas pela Polícia Federal no Estado do Rio Grande do Sul

A Cannabis sativa L. (canábis) contém centenas de substâncias químicas em diferentes classes, contudo, a classe dos canabinoides é encontrada unicamente nesta espécie. Popularmente conhecida como maconha, a canábis é a droga ilícita mais consumida no mundo, sendo o Δ9-tetrahidrocanabinol (Δ9-THC), o principal canabinoide de interesse toxicológico, por ser o responsável pela maioria dos efeitos psicotomiméticos ocasionados e estar associado ao uso abusivo da planta. De outro modo, há alguns anos, vem crescendo significativamente o interesse em pesquisas científicas com a canábis, devido aos importantes efeitos terapêuticos que alguns canabinoides têm apresentado. O canabidiol (CBD), por exemplo, é um canabinoide que além de modular os efeitos eufóricos do Δ9-THC, tem apresentado importantes atividades farmacológicas. Embora haja um crescente aumento nas toneladas de apreensões de canábis realizadas no Brasil, não existem dados que caracterizem o perfil químico e a potência da droga que é utilizada ilegalmente no país. Outro aspecto singular com relação às apreensões de canábis refere-se à nova forma de tráfico internacional de drogas, que vem crescendo significativamente no país. Trata-se da remessa de sementes de canábis em pequenas quantidades através de empresas de transporte. Desta forma, este estudo foi realizado a partir de sementes de canábis apreendidas pela Superintendência de Polícia Federal no Rio Grande do Sul. As sementes foram germinadas e as plântulas foram cultivadas, secas e analizadas, sendo todos os passos em condições controladas. As metodologias analíticas utilizadas foram: espectroscopia no infravermelho próximo (NIRS), espectrometria de massas por ressonância ciclotrônica de íons com transformada de Fourier (FT-ICR MS) e cromatografia à gás com detector de massas (CG/EM). O elevado número de compostos presente nas plantas torna os dados obtidos muito complexos, onde pequenas diferenças entre amostras podem ser negligenciadas. Por isso, foram utilizadas as seguintes ferramentas quimiométricas: HCA, PCA, PLS-DA e SVM-DA. As diferentes metodologias de análise utilizadas neste trabalho, permitiram a análise de um total de 73 amostras de canábis que forneceram dados sob químicos, bem como aspectos distintos e relevantes em toxicologia. As informações obtidas por NIRS associadas às ferramentas quimiométricas HCA e PCA possibilitaram agrupar 29 amostras de canábis em diferentes períodos de crescimento; e as ferramentas quimiométricas PLS-DA e SVM-DA permitiram classificar 29 amostras de canábis em diferentes períodos de crescimento com 100% de acerto. O altíssimo poder de resolução da FT-ICR MS possibilitou a identificação 123 compostos canabinoides e metabólitos a partir das análises realizadas em 73 amostras de canábis, o que resultou na compilação de um abrangente perfil químico de canabinoides, até então não obtido. A eficiência de separação da CG acoplada à identificação dos compostos utilizando EM tornou possível a diferenciação de 73 amostras de canábis em tipo fibra ou tipo droga, além da caracterização do perfil químico de canabinoides e terpenoides presentes nas amostras, obtido pela análise em CG/EM. Desta forma, os dados gerados com o desenvolvimento desta tese possibilitaram a identificação e o conhecimento do perfil químico de amostras de canábis apreendidas no Brasil, apresentando informações até então desconhecidas pela polícia brasileira, bem como pelos profissionais da saúde, comunidade científica, órgãos governamentais e políticos do país e também para a população em geral.Cannabis sativa L. (cannabis) contains hundreds of different classes of chemicals, however, the class of cannabinoids is found only in this plant. Popularly known marijuana, cannabis is the most used illicit drug worldwide, and the Δ9-tetrahydrocannabinol (Δ9-THC), the main cannabinoid regarding toxicological interest, being responsible by the majority of psychotomimetic effects and it is associated with the abuse of the plant. Otherwise, for a few years has grown significantly the interest and scientific research with cannabis due to significant therapeutic effects that certain cannabinoids have shown. The cannabidiol (CBD), for example, is a cannabinoid which in addition to modulate the euphoric effects of Δ9-THC, it has shown significant pharmacological activity. Although there is a growing increase in tons of cannabis seizures conducted in Brazil, there are no data which characterize the chemical profile and potency of the drug that is illegally used in the country. Another unique aspect regarding cannabis seizures refers to the new form of international drug trafficking that has grown significantly in the country. This is the shipment of cannabis seeds in small quantities by transportation companies. Thus, this study was based on samples from cannabis seeds seized by the Superintendence of the Federal Police in Rio Grande do Sul. The seeds were germinated and the seedlings were grown, dried and analyzed, all of the steps under controlled conditions. The analytical methods used were: near infrared spectroscopy (NIRS), Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) and gas chromatography coupled with mass spectrometry (GC/MS). The high number of compounds presente in plants renders them very complex data set, where small differences between samples may be wasted. Therefore, the following chemometric tools were used: HCA, PCA, PLS-DA and DA-SVM. The different methods of analysis used in this study allowed the analysis of a total of 73 samples of cannabis that provided data on chemical and distinct and relevant aspects in toxicology. Information obtained by NIRS chemometric tools associated with HCA and PCA group allowed 29 samples of cannabis in different periods of growth; and chemometric tools PLS-DA and SVM-DA can be classified 29 samples of cannabis in different periods of growth with 100% accuracy. The very high resolution power of FT-ICR MS possible to identify 123 compounds cannabinoids and metabolites from the analyzes carried out on 73 samples of cannabis, which resulted in the compilation of a comprehensive chemical profile of cannabinoids, has not been surpassed. The GC separation efficiency coupled with the identification of compounds using MS made it possible to differentiate 73 cannabis samples in type or fiber type drug, besides the characterization of the chemical profile of cannabinoids and terpenoids present in the samples obtained through the analysis on GC/MS. Thus, the data generated with the development of this thesis allowed the identification and knowledge of the chemical profile of cannabis samples seized in Brazil, presenting information previously unknown to Brazilian police as well as by health professionals, the scientific community, government agencies and politicians of the country and also to the general population

Tuberculous Meningitis: a case report of a late diagnosis

Introduction: Tuberculous meningitis (TM) is the most severe form of tuberculosis, it has high morbidity and mortality. Once it is diagnosed early, treatment response is excellent. Objective: To report and discuss a case of delayed diagnosis of TM. Methodology: It was selected a case from University Hospital of Federal University of Santa Catarina, the analysis of medical records was performed and discussed based on the scientific literature. Results: It was reported the case of a patient that was diagnosed with leukocytoclasticvasculitis (LV) for six months and was admitted presenting granulomatous lung injury, weight loss, fever and dry cough for eigth months. The delay in the diagnosis of tuberculosis (TB) allowed an evolution to TM and the late onset of treatment impaired the patient’s prognosis. It was discussed the vasculitis as a first symptom of tuberculosis, which can occur by deposition of immune complexesformed by antibodies against antigens of bacilli in the vascular wall. Conclusions: Previous diagnosis of LV deviated the focus of the case for their cause, and suggested other pathologic conditions, which contributed to delay in the diagnosis of TB. Despite the late diagnosis and treatment, the therapeutic approaches used for vasculitis, TB and TM followed the protocols described in the literature

Biotransformation of ephedrine by whole cells of Cunnighamella elegans

Ephedrine biotransformation reactions were performed using strain Cunninghamella elegans in order to obtain products similar to human metabolism products. The choice of strains used was based on previous literature review which identified the species Cunninghamella elegans as promising due to their phenotypic characteristics, such as expressing Oxidoreductases enzymes that are necessary for carrying out the oxidation and reduction reactions in the side chain of the molecule ephedrine, in addition to expressing monooxygenases enzymes cytochrome P450 superfamily, which in humans were found in the lipid bilayer of hepatocytes and were responsible for the hydroxylation process of substances. Therefore, it was performed experiments with strains of Cunninghamella elegans acquired by the mycology collection of the Federal University of Pernambuco, originating from different ecosystems. From screening step, it selected URM 4428 strain to present better profile biocatalytic to ephedrine. The results demonstrate the gradual formation of bioproducts in reversible character, and the reduction of bioproducts accompanied by the recovery of the substrate. To our knowledge, there are not previous scientific literature focusing the biotransformation of ephedrine by fungi, which demonstrates the uniqueness specific to our research and presents C. elegans , as a promising auxiliary tool in the exploration and understanding of the metabolism of amphetamine- type drugs in humans