Environmental monitoring is becoming increasingly important, primarily in urban areas due to the concentrated levels of human activities. The air sampling device presented is a novel method to sample air which harnesses the power of paper spray ionization paired with the intrinsic advantages of mass spectrometry such as high sensitivity, high selectivity, high throughput, and the ability to monitor multiple compounds at once.

Made available in DSpace on 2018-08-01T21:58:48Z (GMT). No. of bitstreams: 1 tese_10611_Flavia Tosato.pdf: 3350639 bytes, checksum: 70973978751713caf9a8020e18b13280 (MD5) Previous issue date: 2016-12-09 / Análises rápidas, baratas e eficientes são necessárias para as rotinas laboratoriais, principalmente quando aplicada em química forense. As utilizações de fontes ambientes permitem que as análises sejam diretas e rápidas. Assim, são opções inovadoras para melhorar a sensibilidade e seletividade. Neste trabalho, foi desenvolvida uma metodologia utilizando a espectrometria de massas ambiente por paper spray ionization mass spectrometry (PS-MS) para diferentes matrizes de dados: (i) identificar e quantificar cocaína e seus adulterantes a partir da cromatografia em camada delgada acoplada ao PS-MS; (ii) realizar a discriminação ou classificação de dez marcas brasileiras de cervejas em pilsen e lager; e (iii) identificar a falsificação controlada de Uisque com cachaça visando quantificar o volume de cachaça adicionado na mistura. Na análise de cocaína e adulterantes, os padrões foram identificados na CCD e analisados por PS-MS, e uma curva de calibração foi otimizada. A cerveja foi previamente fracionada usando a metodologia de extração em fase sólida (SPE) com água/metanol e os quatro extratos foram caracterizados no modo positivo de ionização, PS(+)MS. Para ambos os casos, foram aplicados de 20 µL do analito sobre a superfície do papel do PS-MS. Na quantificação de cachaça em misturas de whisk-cachaça. Os resultados indicam que, utilização da CCD acoplado ao PS-MS possibilita a identificação de cocaína e seus adulterantes uma ótima linearidade paras as curvas foi encontrada (cocaína (R2 = 0,9989), benzocaína, (R2 = 0,9934), a cafeína (R2 = 0,9988), a fenacetina (R2 = 0,9966), e a lidocaína (R2 = 0,9987)) e LOD inferior ao da técnica de CCD. Para as amostras de cerveja os sinais de açúcares se sobressaem sobre os demais grupos, e após a extração com água/metanol foi possível distinguir os espectros das diferentes marcas de cervejas e classifica-las em pilsen e lager pela técnica de PCA. Utilizando o PS(±)-MS e PLS com seleção de variáveis foi possível estimar quantitativamente o teor de cachaça misturado ao uísque. Com isso, utilização de fontes ambientes, como PS mostra-se eficaz para identificação de diversas matrizes, facilitando as análises de rotina.

Made available in DSpace on 2018-08-01T21:58:59Z (GMT). No. of bitstreams: 1 tese_11518_Tese_eloilson_Domingos_VERSÃO IMPRESSÃO.pdf: 3568693 bytes, checksum: cd4564d6013c45b3ddc32e2ccd0a7957 (MD5) Previous issue date: 2017-11-30 / Com o advento de uma nova família de técnicas de ionização, a espectrometria de massas (MS) ambiente foi introduzida como uma maneira simples de gerar íons na MS. Entre elas, a técnica de ionização por paper spray (PS) demonstrou-se versátil para resolver inúmeros problemas. Neste trabalho, a técnica de PS-MS no modo de ionização positiva (PS(+)-MS) foi aplicada em três subáreas da química forense: drogas de abuso e documentoscopia e ervas fitoterápicas. Na primeira, a técnica PS-(+)-MS determinou o perfil químico de drogas sintéticas tais como selos contendo 25I-NBOMe (2-(4-iodo-2,5-dimetoxifenil)-n-[(2-Metoxifenil) metil] etamina), extratos e folhas de canabinóides naturais (&#916;9-THC) e canabinóides sintéticos (JWH-073; 5F-AKB48; JWH-122; JWH-250; e JWH-210). Em todos os casos, as moléculas foram facilmente detectadas na sua forma protonada ([M+H]+) com resolução, m/&#916;m50% = 67,000 e uma precisão de massa menor que <2 ppm. O desempenho da técnica PS(+)-MS foi comparado com o de outras fontes de ionização: leaf Spray (LS-MS) e eletrospray (ESI-MS). Além disso, um método foi desenvolvido para quantificar uma série de drogas ilícitas (de 1 a 110 &#120583;g L-1, com linearidade de R2> 0,99). Em documentoscopia, a técnica PS(+)-MS foi capaz de obter com sucesso o perfil químico de diferentes canetas vendidas no comercio. A intensidade relativa (RII372) do corante azul de metileno foi monitorada para discriminar os cruzamentos de traços de canetas e, até a data, os documentos questionados. O perfil químico das notas brasileiras de segunda geração de R$ 100,00 (reais) também foi explorado. A técnica PS(+)-MS foi capaz de classificar as notas em autêntico ou falsificadas. Constatando que amostras falsas possuem um perfil químico similar às notas caseiras da impressora Deskjet®. Finalmente, aplicamos PS(+)-MS para verificar a autenticidade de ervas usadas para perda de peso. Quinze diferentes medicamentos à base destas ervas foram analisados por PS(+)-MS. Encontrando diferentes tipos de compostos: sibutramina (m/z 280), cafeína (m/z 195), glicose (m/z 203 [M+Na], m/z 219 [M+K] e sacarose (m/z 381 [M+K]). Todas essas substâncias não estavam listadas em seus rótulos. Em geral, a técnica PS(+)-MS mostrou ser uma excelente ferramenta analítica em química forense. O segundo estudo abordado foi o controle de qualidade de combustíveis com aplicação do micro-espectrômetro portátil na região do infravermelho próximo (MICRONIR). Neste trabalho, verificou-se a potencialidade do MICRONIR no controle da qualidade do combustível. Assim, modelos quimiométricos (usando o PLS - regressão por mínimos quadrados parciais) foram desenvolvidos para quantificar biodiesel em Diesel (LD=0,5 wt%; LQ=1,8 wt%, e RMSEP=1.8 wt%); enxofre em Diesel (LD=2,4 mg L-1; LQ=8,0 mg L-1, e RMSEP=13,2 mg L-1); gasolina, etanol e metanol em gasolina tipo C (LDgas=0,55 wt%; LQgas=1,84 wt%, e RMSEPgas=0,81 wt%; LDet=0,75 wt%; LQgas=2,50 wt%, e RMSEPeth=3,81 wt%; LDmet = 0,85 wt%; LQmet = 2,84 wt%, e RMSEPmet = 1,80 wt%); e água, metanol e etanol em etanol combustível hidratado (LDH2O = 0,04 wt%; LQH2O=1,29 wt%, e RMSEPH2O=1,05 wt%; LDmet= 0,52 wt%; LQmet = 1,73 wt%, e RMSEPmet =2,78wt %; LDet=1,22 wt%; LQet=4,07 wt%, e RMSEPet= 4,41 wt%). O método proposto apresentou alta precisão e exatidão em todos os casos, e os resultados mostraram que a técnica MICRONIR teve excelente desempenho no controle da qualidade do combustível.

Indiana University-Purdue University Indianapolis (IUPUI) / Paper spray mass spectrometry is an alternative technique for toxicological screening that is able to quickly and adequately screen for compounds encountered in postmortem investigations with little sample handling and no sample preparation. For analysis of dried blood spots using a triple quadrupole mass spectrometer, detection criteria were defined to align with relevant regulatory guidelines while considering how fragment ion selection, method sensitivity, and fragment ion ratio tolerances are best utilized in paper spray mass spectrometry. For analysis, drugs and drug metabolites relevant to postmortem investigations were spiked into drug-free blood, and by monitoring two fragment ion channels in selected reaction monitoring mode, as well as the ratio between the two fragment ions, a method was developed capable of detecting over 120 drug and drug metabolites at concentrations relevant to postmortem drug screening. Total analysis time for the developed method is less than 8 minutes, and less than 50µL of sample and 5mL of solvent are consumed during analysis.

The research described in this dissertation is divided into two sections. The first section focuses on mass spectrometry-based bottom-up proteomics application to identify fungal protein biomarkers of invasive aspergillosis infection. The second part focuses on instrument development to improve current ionization and dissociation technologies for characterizing topology and substructure of protein complexes. Part I of this dissertation describes the identification and validation of protein biomarkers for Invasive Aspergillosis (IA), a fatal pulmonary infection. Aspergillus fumigatus, the organism responsible for this disease, is an opportunistic fungus. Immunocompromised individuals can suffer from IA due to impaired immune response. The current diagnostic tools are time-consuming and have variable sensitivity and specificity. Hence, treatments for IA are often administered too late. The goal of this research is to use mass spectrometry to identify and validate novel fungal protein biomarkers for IA. To tackle this challenge, several systems were studied. Commercial Aspergillus antigen was used for method development, and to serve as standards for spiking and comparison. Mouse models of different disease manifestations were used in the initial study to compare proteomic differences in carefully controlled disease states. Although it was not successful in providing candidate biomarkers, the mouse samples provided host response protein data. Human patient samples yielded the most promising results. Several Aspergillus proteins have been identified and validated from patient bronchoalveolar lavage fluid, and could have the potential to be later used on a diagnostic platform. Part II describes two instrument development projects: incorporation of a surface-induced dissociation device into a commercial ion mobility time-of-flight mass spectrometer, and the development of a paper spray ionization source. Protein complexes are often studied using collision-induced dissociation (CID), which does not provide enough substructure information. Surface-induced dissociation (SID) allows access to higher energy fragmentation pathways, which generates more useful substructure information. Its potential is demonstrated with three systems here-- one metal cluster and two protein complexes. All systems show that SID can provide more useful structural information than CID under similar conditions. The development of a paper spray (PS) source for protein complex ionization provides another way to study protein complexes. Chapter 9 shows that this ionization method can also be applied to protein complexes. Under the same conditions as its nanospray counterpart, similar mass spectra can be obtained using PS. This exciting result is the first demonstrations that PS can be used for protein complexes while maintaining each protein complex's native structure and conformation.

Indiana University-Purdue University Indianapolis (IUPUI) / In this work we developed paper spray mass spectrometry methods to obtain lower detection limits for pharmaceuticals and drugs of abuse. The second chapter investigates blood fractionation membranes for their ability to obtain lysis free plasma from whole blood without changing the drug concentration relative to centrifugation. We presented a device capable of obtaining and analyzing plasma samples from whole blood and obtaining quantitative results similar to traditional methods. In the third chapter the properties of the paper substrate are investigated systematically for their impacts on ionization efficiency and recovery in combination with the solvent choice. The fourth and fifth chapters detail a simple method for lowering detection limits using a method called paper strip extraction. In this method biofluids are wicked through either sesame seed oil or solid phase extraction powder on a paper strip to concentrate and preserve (in the case of THC) analytes out of biofluids. The use of 3D printing for rapid prototyping and how it potentially impacts paper spray MS sensitivity is outlined in the final chapter.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2015-11-19T11:42:47Z No. of bitstreams: 2 Dissertação - Thays Colletes de Carvalho - 2015.pdf: 2499571 bytes, checksum: fe876dabf52c3d28675aeb6e1dae2d21 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2015-11-19T11:44:31Z (GMT) No. of bitstreams: 2 Dissertação - Thays Colletes de Carvalho - 2015.pdf: 2499571 bytes, checksum: fe876dabf52c3d28675aeb6e1dae2d21 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2015-11-19T11:44:31Z (GMT). No. of bitstreams: 2 Dissertação - Thays Colletes de Carvalho - 2015.pdf: 2499571 bytes, checksum: fe876dabf52c3d28675aeb6e1dae2d21 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-03-06 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / 2G Ethanol production is a laborious process that may result in formation of glucose -fermenting yeasts, which reduces the process yield. The search for new rapid and sensitive methods are required to show the condition with the formation of free sugars and absence of fermentation inhibitors (Hidroximetilfurfural and furfural). The present work proposes utilizing the Paper Spray (PS) Ionization source as a method of detecting and quantifying fermentation yeasts inhibitors and sugars generated in 2G ethanol-acquiring processes. The method was optimized and was obtained as calibration curves for the glucose obtained by enzymatic hydrolysis process. These conditions were used for analysis actual sample of sugarcane bagasse liquors. The best analysis conditions by PS were: distance between 1 and 5 mm, dry spot application mode, Whatman Grade 1 paper, 60º angle, modified paper with paraffin barrier, 10 μL volume and sample application concentration of 0,1mg/mL. The detection of the expected product in the pretreatment and hydrolysis liquors of sugarcane bagasse was performed as expected. The paper with paraffin channel proved to be more effective for the quantification of glucose with improved linearity (0.9952) and limits of detection (2.77 mmol/L) and quantification (9.27 mmol/L) compared to without changing paper and paper cur ends. Applying the PS prototype in determining sugars and inhibitors in sugarcane bagasse liquors has proven to be efficient and quick, opening up a perspective of utilizing this method on monitoring Ethanol 2G production processes. / A produção de etanol 2G é um processo laborioso, que pode resultar na formação de inibidores de leveduras fermentadoras de glicose, o que diminui os rendimentos do processo. A busca de novas metodologias rápidas e sensíveis são necessárias para evidenciar a condição com a formação de açúcares livres e ausência de inibidores da fermentação (Hidroximetilfurfural e furfural). Este trabalho apresenta a proposta de utilizar a fonte de ionização Paper Spray Ionization (PS) como método de detecção e quantificação de inibidores e açúcares gerados nos processos de obtenção de etanol 2G. O método foi otimizado e obteve-se curvas de calibração para a glicose obtida pelo processo de hidrolise enzimática. Essas condições foram usadas para análise de amostras reais de licores de bagaço de cana-de-açúcar. As melhores condições de análise por PS foram: Distância entre 1 e 5 mm, modo de aplicação por aplicação a seco, papel Whatman Grade 1, ângulo de 60°, papel modificado com barreira de parafina, volume de 10 μL e concentração de amostra de 0,1mg/mL. A detecção dos produtos esperados nos licores de pré-tratamento e hidrólise do bagaço de cana-de-açúcar foi realizada como esperado. O papel com canal de parafina mostrou-se ser mais eficaz para a quantificação da glicose com melhor linearidade (0,9952) e limites de detecção (2,77 mmol/L) e quantificação (9,27 mmol/L) em relação ao papel sem modificação e o papel arredondado. A aplicação do PS na determinação de açúcares e inibidores em licores de bagaço de cana-deaçúcar mostrou-se eficiente e rápido abrindo a perspectiva de utilização deste método no acompanhamento de processos de produção de etanol 2G.

Submitted by Franciele Moreira (francielemoreyra@gmail.com) on 2018-04-04T11:53:17Z No. of bitstreams: 2 Dissertação - Marcella Ferreira Rodrigues - 2018.pdf: 1869503 bytes, checksum: 8b0fc83d8e1366a8a5952aa48164bfd3 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-04-04T13:25:36Z (GMT) No. of bitstreams: 2 Dissertação - Marcella Ferreira Rodrigues - 2018.pdf: 1869503 bytes, checksum: 8b0fc83d8e1366a8a5952aa48164bfd3 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-04-04T13:25:36Z (GMT). No. of bitstreams: 2 Dissertação - Marcella Ferreira Rodrigues - 2018.pdf: 1869503 bytes, checksum: 8b0fc83d8e1366a8a5952aa48164bfd3 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-02-28 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Water is an essential natural resource for life that is compromised by the release of emerging contaminants, including synthetic hormones. These compounds can be released directly through industrial effluents and indirectly through domestic sewage. Monitoring emergent contaminants (ECs) in the aquatic environment has become a priority as they pose risks to human health and the environment because of their ability to deregulate the endocrine system. Thus, verification analyzes of the emerging contaminants produced by the pharmaceutical industries are required. The most common analytical methods for analysis of this class of compounds involve the use of liquid or gas chromatography coupled to tandem mass spectrometry (MS / MS) with procedures for the preparation of long samples and long chromatographic runs. In this study, a methodology was developed for the analysis of emerging contaminants in raw and treated industrial wastewater from a local industry's hormone production line by an environmental ionization technique known as Paper Spray Ionization (PSI). Here, we have proposed an analytical approach based on the use of waxed papers. Analyzes of the synthetic hormones levonorgestrel and algestone acetophenide occurred by adding the sample to a triangular paper that was moistened with methanol solution (0.1% formic acid). The paper was fixed at the input of the mass spectrometer through a metal clip which provided a voltage to the paper. The analytical curves presented a linear correlation coefficient above 0.99. For the hormone levonorgestrel the intra-day and inter-day precision ranged from 0.5 to 5.3%, accuracy ranged from -7.0 to 4.2% while recovery ranged from 82.2 to 101.3%. For the hormone algestone acetophenide the intra-day and inter-day precision ranged from 0.9 to 10.7%, accuracy ranged from -7.7 to 8.9% while recovery ranged from 88.4 to 101.6%. The levonorgestrel hormone was also analyzed with the classic LC-MS method, showing consistent results between the two approaches, thus confirming the reliability of the proposed method. / A água é um recurso natural essencial para vida que ultimamente está comprometido pela descarga de contaminantes emergentes, dentre os quais, hormônios sintéticos. Esses compostos podem ser lançados diretamente através de efluentes industriais e indiretamente através de esgoto doméstico. O monitoramento de contaminantes emergentes (ECs) no ambiente aquático tem se tornado uma prioridade, visto que eles apresentam riscos para a saúde humana e para meio ambiente devido a sua capacidade de desregular o sistema endócrino. Dessa forma, é necessário análises de verificação dos contaminantes emergentes produzidos pelas indústrias farmacêuticas. Os métodos analíticos mais comuns para análises dessa classe de compostos envolvem o uso de cromatografia líquida ou gasosa, acoplado a espectrometria de massas tandem (MS/MS) com procedimentos de preparos de amostras demorados e longas corridas cromatográficas. Neste trabalho desenvolveu-se uma metodologia para análise de contaminantes emergentes em água residual industrial bruta e tratada de uma linha de produção de hormônios de uma indústria local, por uma técnica de ionização ambiente conhecida como Paper Spray Ionization (PSI), cuja principal vantagem é o preparo mínimo de amostra. Neste trabalho, a técnica de PSI com papel contendo barreiras de parafinas foi empregada para análise de hormônios em águas residuárias. As análises dos hormônios sintéticos levonorgestrel e algestona acetofenida ocorreram através da adição da amostra sobre um papel triangular que foi umedecido com solução de metanol (0,1% de ácido fórmico). O papel foi fixado na entrada do espectrômetro de massas através de um clipe metálico que forneceu uma voltagem ao papel. As curvas analíticas apresentaram coeficiente de correlação linear acima de 0,99. Para o hormônio levonorgestrel a precisão intra-dias e inter-dias variou de 0,5 a 5,3%, a exatidão variou de -7,0 a 4,2% enquanto a recuperação variou de 82,2 a 101,3%. Para o hormônio algestona acetofenida a precisão intra-dias e inter-dias variou de 0,9 a 10,7%, a exatidão variou de -7,7 a 8,9% enquanto a recuperação variou de 88,4 a 101,6%. O hormônio levonorgestrel também foi analisado com o método clássico LC-MS, mostrando resultados consistentes entre as duas abordagens, confirmando assim a confiabilidade do método proposto.

<p>Suboptimal dosing of anti-microbial agents increases the likelihood of therapeutic failure and resistance. Dosing optimization, while an attractive approach to combat these issues, is difficult to implement due to the different pharmacokinetics of each individual. These limitations highlight the inadequacies of a “standardized” dosing strategy. Therapeutic drug monitoring (TDM) provides a tailored treatment for individuals while avoiding adverse side effects from compounds with a narrow therapeutic window where elevated concentrations of a drug cause organ toxicity. This strategy involves accurately measuring the concentration of the analyte and interpreting the results based on pharmacokinetic parameters. Clinicians then draw conclusions regarding dose adjustment for their patient. However, TDM is expensive and difficult to perform because measurements occur in biofluids. Rapid and robust methods are necessary to quantify antimicrobial agents at the institutional level to guide patient care toward improved outcomes in serious infection. Paper spray ionization (PS), an emerging ambient ionization technique for clinical settings, demonstrations a wide versatility both in analyte variety and applications. This technique offers a rapid, accurate method to analyze these compounds with low rates of false positives even when multiplexing.</p><p><br></p><p>The work herein explains the method development of assays for TDM of various antimicrobial agents. Chapters two and three describe ways to improve the quantitative capability of paper spray through substrate pre-treatment, modification, and manipulation of key factors. Chapter four describes real-world applications for paper spray utility in clinical settings with the cross-validation of antifungal agents against a “gold standard” method. The final chapter, while not clinical based, describes the method development process for a LC-MS/MS assay to detect urobilinoids in fly guts.</p>

Indiana University-Purdue University Indianapolis (IUPUI) / Suboptimal dosing of anti-microbial agents increases the likelihood of therapeutic failure and resistance. Dosing optimization, while an attractive approach to combat these issues, is difficult to implement due to the different pharmacokinetics of each individual. These limitations highlight the inadequacies of a “standardized” dosing strategy. Therapeutic drug monitoring (TDM) provides a tailored treatment for individuals while avoiding adverse side effects from compounds with a narrow therapeutic window where elevated concentrations of a drug cause organ toxicity. This strategy involves accurately measuring the concentration of the analyte and interpreting the results based on pharmacokinetic parameters. Clinicians then draw conclusions regarding dose adjustment for their patient. However, TDM is expensive and difficult to perform because measurements occur in biofluids. Rapid and robust methods are necessary to quantify antimicrobial agents at the institutional level to guide patient care toward improved outcomes in serious infection. Paper spray ionization (PS), an emerging ambient ionization technique for clinical settings, demonstrations a wide versatility both in analyte variety and applications. This technique offers a rapid, accurate method to analyze these compounds with low rates of false positives even when multiplexing. The work herein explains the method development of assays for TDM of various antimicrobial agents. Chapters two and three describe ways to improve the quantitative capability of paper spray through substrate pre-treatment, modification, and manipulation of key factors. Chapter four describes real-world applications for paper spray utility in clinical settings with the cross-validation of antifungal agents against a “gold standard” method. The final chapter, while not clinical based, describes the method development process for a LC-MS/MS assay to detect urobilinoids in fly guts.