O consumo dos diferentes tipos de drogas está associado a problemas sociais, econômicos e de saúde pública, em todas as regiões no mundo. Dentre os problemas de saúde pública, podemos destacar a alta prevalência de comorbidade entre o uso de drogas e os transtornos mentais. A Cannabis, a cocaína e o crack são as drogas ilícitas mais consumidas no Brasil. A utilização do cabelo como matriz biológica para determinação destas drogas permite avaliar o uso crônicos pelos indivíduos, uma vez que o cabelo é uma matriz estável, de fácil manipulação e a janela de detecção depende apenas do comprimento do cabelo. Entretanto, a análise em cabelo ainda representa um desafio analítico. Foram desenvolvidos dois métodos para a detecção das diferentes drogas no cabelo, com diferentes enfoques analíticos. O primeiro método (desenvolvido durante o estágio de doutorado sanduíche na Itália) identificou e quantificou cocaína e metabólitos usando a técnica de column switching e detecção por LC-MS/MS. O segundo método foi desenvolvido para determinação de canabinóides nas amostras de cabelo utilizando GC-MS. O diferencial deste método foi a utilização de um novo dispositivo de extração em fase sólida (as ponteiras DPX) para concentração e purificação do extrato, utilizando menor quantidade de solventes. A determinação dos canabinóides e cocaínicos foi realizada nas amostras de cabelo da população atendida no CAPS - AD de Ribeirão Preto, São Paulo. Além da coleta da amostra de cabelo, o sujeito foi submetido a uma entrevista, onde os seguintes instrumentos de avaliação foram aplicadas: Questionário sobre a saúde do Paciente 9, Inventário de fobia social, Self Report Questionnaire, Questionário de Ansiedade de Beck, Inventário de Depressão de Beck e Questionário sobre o uso da Cannabis, bem como um questionário elaborado pelo pesquisador para coleta de dados sociodemográficos, consumo de substâncias e dados sobre a amostra de cabelo, como comprimento, cor, tintura ou coloração. As amostras de cabelo foram analisadas e a média das concentrações de cada droga encontrada no cabelo foi correlacionada com os indicadores clínicos de transtorno mentais, obtidos através dos instrumentos de avaliação psiquiátrica. A maior prevalência de indicadores clínicos positivos para transtornos psiquiátricos entre a população estudada foi de transtornos mentais comuns, entre eles a ansiedade e depressão. A comparação da média de concentração de Cannabis, cocaína e crack no cabelo com os indicadores clínicos positivos para os transtornos não apresentou resultados estatisticamente significantes. Entretanto, podemos inferir que os sujeitos que apresentaram maior concentração média de Cannabis e cocaína no cabelo possuíam mais indicadores clínicos positivos para sintomas mentais comuns e depressão maior enquanto que os sujeitos usuários de crack possuíam mais indicadores clínicos positivos para sintomas ansiedade. Apesar de algumas limitações, podemos concluir que o estudo possibilitou estimar a prevalência da morbidade entre abuso de drogas ilícitas e transtornos psiquiátricos na população atendida no Centro de Atenção Psicossocial Álcool e drogas de Ribeirão Preto. / Different kinds of drug use have been associated to social, economic and health public problems worldwide. Among the high prevalent public health problems is the comorbidity between drug abuse and psychiatric disorders. Cannabis, cocaine and crack are the most consumed illicit drugs in Brazil. Hair use as the biological matrix for the determination of these drugs allows to evaluate chronic use, once hair is a stable matrix, easy to manipulate and the window detection only depends on the size of the hair. However, hair analysis still represents an analytical challenge. It was developed two methods for the detection of the drugs in hair, with different analytical approach. The first method (developed during doctoral stage in Italy) had identified and quantified cocaine and metabolites using column switching technique and LCMS/ MS detection. The second method was developed for determination of cannabinoids in hair samples using GC-MS. Decontamination procedure was the same cited above. The differential of this method was the use of a new device in solid phase extraction (DPX tips) for the extracts concentration and purification, using less solvents volumes. Cannabinoids and cocaine analysis were performed in hair samples from people who were enrolled in the CAPS - AD of Ribeirão Preto, São Paulo. Besides hair collection, the individual was submitted to an interview, where it was applied the following evaluation scales: Patient health questionnaire - 9, Social phobia inventory, Self report questionnaire, Beck anxiety inventory, Beck depression inventory and Cannabis research questionnaire as well as a questionnaire made by the author to collect sociodemographic data, substance consume and hair data. Hair samples were analyzed and the concentrations were correlated with positive clinical factors of mental disorders obtained through evaluation scales. The highest prevalence of positive clinical indicators for psychiatric disorders among the population studied was of common mental disorders, including anxiety and depression. The comparison of the average concentration of Cannabis, cocaine and crack in the hair with the positive clinical indicators for the disorders did not show statistically significant results. However, we can infer that the subjects who had higher average concentration of Cannabis and cocaine in hair had showed positive indicators for common mental symptoms and major depression as well as the subjects who had higher average concentration of crack in hair had showed positive indicators for anxiety. Despite of some limitations, we can conclude that the study had allowed estimating the prevalence of morbidity among illicit drugs abuse and psychiatric disorders in the population enrolled in the psychosocial care center in Ribeirão Preto.

Indiana University-Purdue University Indianapolis (IUPUI) / Analyses of tobacco products and their usage are well-researched and have implications in analytical chemistry, forensic science, toxicology, and medicine. As such, analytical methods must be developed to extract compounds of interest from tobacco products and biological specimens in order to determine tobacco exposure. In 2009, R.J. Reynolds Tobacco Co. released a line of dissolvable tobacco products that are marketed as a smoking alternative. The dissolvables were extracted and prepared by ultrasonic extractions, derivatization, and headspace solid phase microextraction (SPME) with analysis by gas chromatography-mass spectrometry (GC-MS). The results show that the compounds present are nicotine, flavoring compounds, humectants and binders. Humectant concentrations vary among different tobacco types depending on the intended use. Humectants were quantified in various tobacco types by GC and “splitting” the column flow between a flame ionization detector (FID) and an MS using a microfluidic splitter in order to gain advantage from the MS’s selectivity. The results demonstrated excellent correlation between FID and MS and show that MS provides a higher level of selectivity and ensures peak purity. Chemometrics was also used to distinguish products by tobacco type. Hair is a common type of evidence in forensic investigations, and it is often subjected to mitochondrial DNA (mtDNA) analysis. Preliminary data was gathered on potential “lifestyle” markers for smoking status as well as any indications of subject age, gender, or race by investigating the organic “waste” produced during a mtDNA extraction procedure. The normally discarded organic fractions were analyzed by GC-MS and various lipids and fatty acids were detected. At this point, a total vaporization-SPME (TV-SPME) method was theorized, developed, and optimized for the specific determination of nicotine and its metabolite, cotinine. The theory of TV-SPME is to completely vaporize an organic extract which will eliminate the partitioning between the sample and the headspace, thereby simplifying the thermodynamic equilibrium. Parameters such as sample volume, incubation temperature, and extraction time were optimized to achieve the maximum analyte signal. Response surface methodology (RSM) is a statistical model that is very useful in predicting and determining optimum values for variables to ensure the ideal response. RSM was used to optimize the technique of TV-SPME for the analysis of nicotine and cotinine. Lastly, quantitation of nicotine and cotinine in human hair typically requires large sample sizes and extensive extraction procedures. Hence, a method using small sample sizes and a simple alkaline digestion followed by TV-SPME-GC-MS has been developed. Hair samples were collected from anonymous volunteers and nicotine and cotinine were identified and quantitated in the hair of tobacco users.

Die Systematische Toxikologische Analyse (STA) stellt auf Grund der großen Vielfalt und der ständigen Zunahme an toxikologisch relevanten Substanzen eine der größten Herausforderungen in der chemischen Analyse dar. In der vorliegenden Arbeit wurde daher die Eignung der Flüssigchromatographie in Kombination mit der Hybrid-Quadrupol-Flugzeitmassenspektrometrie (LC-QTOF-MS) für diesen Zweck untersucht. Dazu wurden eine Datenbank mit über 7360 und eine CID-Spektrenbibliothek mit mehr als 2720 toxikologisch relevanten Substanzen erstellt und geeignete Probenvorbereitungsmethoden entwickelt. Die Erprobung der Methoden erfolgte an dotierten Blut- und Haarproben. Hierbei zeigte sich, dass die Analyse im Auto-MS/MS-Modus (Messzyklen von MS- und MS/MS-Spektren) eine Identifizierung basischer Substanzen mittels CID-Spektren zwischen 0,5 und 2 ng/ml im Blut ermöglichte. Die Nachweisgrenzen der für 24 Wirkstoffe validierten Methode in Haaren lagen bei 3 bis 15 pg/mg. Die Eignung der LC-QTOF-MS zur STA von Haarproben wurde an 30 Drogentodesfällen und 60 Todesfällen mit bekannter chronischer Medikamenteneinnahme zu Lebzeiten sowie an 77 Blutproben nachgewiesen. Für die Suche nach Metaboliten wurde ein Metaboliten-Tool entwickelt. In der praktischen Anwendung auf Datenfiles von Blut- und Haarproben erwies sich das Tool als wertvolles Hilfsmittel zur Identifizierung unbekannter Peaks und zur Bestätigung von Suchergebnissen in der Datenbank. Zur automatischen Konzentrationsabschätzung identifizierter Substanzen wurde ein Tool „Estimate Concentration“ geschaffen. Die Überprüfung des Verfahrens an realen Blut- und Haarproben durch Vergleich mit HPLC-DAD- und GC-MS-Ergebnissen wies eine gute Übereinstimmung der Konzentrationen auf. Insgesamt zeigten die Untersuchungen, dass die LC-QTOF-MS zurzeit die am besten geeignete Methode für die STA darstellt. Auch bei einem erst später aufkommenden Verdacht kann eine gezielte Suche in dem bereits gemessenen Datenfile durchgeführt werden. / Due to the large variety and the steady increase of toxicologically relevant substances, systematic toxicological analysis (STA) is one of the most difficult tasks in analytical chemistry and, therefore, a steady topic of research and methodical improvement. For this reason, the suitability of liquid chromatography in combination with hybrid quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) for STA was investigated. For this purpose, a database of more than 7360 and a CID spectra library of more than 2720 toxicologically relevant substances and suitable methods for sample preparation were developed. The application was evaluated at spiked blood and hair samples. It was found that the analysis in Auto-MS/MS mode (alternating measurement cycles of MS and MS/MS spectra) allowed substance identification in blood using CID spectra between 0.5 and 2 ng/ml for basic substances. The detection limits of the validated method in hair ranged from 3 to 15 pg/mg for 24 drugs. The suitability of LC-QTOF-MS for STA was tested for hair samples from 30 drug-related death cases and from 60 death cases with known chronic medication as well as for 77 blood samples. For the search of metabolites, a metabolite tool was developed. In the practical application to data files from blood and hair samples, the tool proved to be very helpful for identification of unknown peaks and for confirmation of results obtained only from the database without CID spectra. A tool "Estimate Concentration" was created for automatic estimation of concentrations of identified substances. The application to real blood and hair samples and the comparison of the concentrations with results from HPLC-DAD and GC-MS showed good agreement. Overall, these investigations showed that LC-QTOF-MS is currently the most favorable method for STA. Because of the comprehensive registration of all substances in a sample, the data files can be checked for the presence of certain poisons even later without new measurements.