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Rodrigues, Lívia Riberti 1988. "Análise de impurezas de formas farmacêuticas sólidas por MALDI Mass Spectrometry Imaging (MALDI-MSI)." [s.n.], 2014. http://repositorio.unicamp.br/jspui/handle/REPOSIP/312437.
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Orientador: Rodrigo Ramos CatharinoTexto em português e inglês
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas
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Resumo: Atualmente, as doenças cardiovasculares constituem uma das primeiras causas de mortes no Brasil e no mundo. Neste cenário, as estatinas constituem uma notável classe de medicamentos redutores de colesterol e têm sido associadas com uma expressiva diminuição da morbidade e mortalidade cardiovascular para pacientes em prevenção primária ou secundária da doença coronariana. Elas agem inibindo competitivamente a enzima HMG-CoA redutase, através da afinidade destes fármacos pelo sítio ativo da enzima. Esta enzima é responsável por catalisar a conversão do substrato HMG-CoA em mevalonato, um dos precursores do colesterol. A crescente necessidade e busca por medicamentos cada vez mais efetivos traz a preocupação na segurança destes produtos para seus usuários. Neste sentido, o conhecimento das impurezas e produtos de degradação torna-se necessário para garantir sua qualidade. Uma técnica muito utilizada para análises de impurezas e degradantes é a espectrometria de massas, pois é uma técnica sensível e seletiva e permite elucidar as estruturas químicas presentes na formulação do medicamento. Sendo assim, amostras de Atorvastatina cálcica foram analisadas pela técnica de espectrometria de massas por imagem (MALDI-MSI), permitindo a quantificação de impurezas do medicamento através da imagem da distribuição dessa impureza no comprimido. Dessa forma, é possível minimizar o preparo de amostra e obter um melhor conhecimento da formulação
Abstract: Currently, cardiovascular diseases constitute one of the first causes of deaths in Brazil and in the world. In this scenario, the statins are a notable class of medicines and cholesterol reducers have been associated with a significant reduction in cardiovascular morbidity and mortality for patients in primary or secondary prevention of coronary heart disease. They act by inhibiting competitively the enzyme HMG-CoA reductase, through the affinity of these drugs by the active site of the enzyme. This enzyme is responsible for catalyzing the conversion of HMG-CoA to mevalonate substrate, one of the precursors of cholesterol. The growing need and search for increasingly effective drugs brings the concern on the safety of these drugs for their users. In this sense, the knowledge of the impurities and degradation products becomes necessary to ensure their quality. A widely used technique for analysis of impurities and degrading is mass spectrometry, because it is a sensitive and selective technique and allows elucidating the chemical structures of the present formulation of the medicinal product. Thus, samples of Atorvastatin calcium were analyzed by the technique of mass spectrometry imaging (MALDI-MSI), which allows the quantification of impurities from the medicine through the image of the distribution of impurity in the tablet. That way, it is possible minimize sample preparation and get a better understanding of the formulation
Mestrado
Ciencias Biomedicas
Mestra em Ciências Médicas
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Lai-Rowcroft, Lindsay Ling Gi. "Novel surfaces for MALDI-MS." Thesis, University of Manchester, 2013. https://www.research.manchester.ac.uk/portal/en/theses/novel-surfaces-for-maldims(331dd97a-881e-4ed0-908f-d5947f3ebeba).html.
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Matrix assisted laser desorption/ionisation mass spectrometry (MALDI-MS) for small molecule analysis has been plagued with inherent problems associated with matrix interference. The matrix plays an important role in MALDI-MS where it has the ability to absorb UV energy from the laser employed and transfer it to the analyte, acts as a proton donor and protecting the analytes from being obliterated. For decades, research has been performed to eradicate matrix interference by matrix avoidance, finding alternative matrices, suppression through sample preparation methods and via chemical modification.In this investigation a number of the above mentioned approaches have been undertaken. First, a mesoporous silica powder, SBA-16, functionalised with a phenyl group to absorb UV from the MALDI-MS laser gave unfruitful results due to inhomogeneous dispersion of the SBA-16 powder. Therefore the same material was prepared but as a thin film and a homogeneously coated surface was generated with the phenyl group incorporated into the silica and this was compared with a conventional matrix, 2,5-dihydroxybenzoic acid (DHB). This was by far the most sensitive method which was accurate, with little background noise and importantly for small molecule analysis clear of matrix interference. Other surface systems were also tested such as graphene on copper and silver on copper, but the functionalised SBA-16 thin film remained the best. Graphite and 2B pencil were also investigated for MALDI-MS but were compared with conventional matrices (DHB and α-cyano-4-hydroxycinnamic acid (CHCA)) in a functional genomics study. The ability of all methods to find subtle phenotypic differences in various yeast strains was assessed with the help of multivariate data analysis (MVDA). Although DHB came out best, 2B pencil produce notably good separations that correlated nicely with the different genotypes. Therefore in addition to conventional matrices, 2B pencil should be considered for functional genomic studies when MALDI-MS is used as it is such a rapid and inexpensive method. Finally, chemical modifications were performed on amino acids where picolinic acid was used to attach a chromophore to the compounds, therefore, allowing UV absorption from the laser. Upon attaching the picolinate UV absorbing group, the amino acid compounds were detected LC-MS at an increased intensity of 10 to 100-fold. Moreover, enhanced separation in LC-MS was also observed.This project has successfully investigated alternative approaches to matrix-free MALDI-MS analysis. Functionalised SBA-16 thin films were by far the best method and this novel surface for MALDI-MS has the potential to transform small molecule analysis.
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Tandina, Fatalmoudou. "Mise au point et application de technologies innovantes pour l'étude des moustiques, de leur préférence trophique et de leur microbiote." Thesis, Aix-Marseille, 2018. http://www.theses.fr/2018AIXM0277/document.
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Les moustiques sont les principaux vecteurs incriminés dans la transmission d’agents pathogènes à l’homme. L’identification précise des espèces de moustiques est importante pour distinguer les espèces vectrices des non vectrices. La détermination de l’origine du repas sanguin des moustiques vecteurs est indispensable dans la compréhension du comportement des espèces vectrices. Nous avons mise à jour la littérature actuelle sur la faune Culicidienne du Mali. Ainsi, nous avons listé 106 espèces de moustiques actuellement enregistrée au Mali dont 28 Anophelinae et 78 Culicinae. Nous avons ensuite évalué l’efficacité du MALDI-TOF MS à identifier des moustiques collectés au Mali et déterminer leur source de repas sanguin. Nous avons confirmé la robustesse du MALDI-TOF MS à identifier un grand nombre de sang d’animaux. Nous avons artificiellement gorgé des femelles de An. gambiae et An. coluzzii sur différents types de sang d’animaux. Nous avons obtenu 100% d'identification correcte du repas de sang pour les spécimens collectés 1h à 24h après le gorgement. Ensuite nous avons expérimentalement gorgés An. gambiae, An. coluzzii et Ae. albopictus sur des repas de sang successif et mixte par MALDI-TOF MS. Nos résultats révèlent que le MALDI-TOF MS est tout à fait capable d’identifier le repas mixte. Mais en ce qui concerne le repas successif seul le dernier repas de sang est identifié. Enfin nous avons utilisé la culturomique et le MALDI-TOF pour l’étude du microbiote digestif de moustiques collectés sur le terrain au Mali et à Marseille. Cette approche a révélé une grande diversité du microbiote digestif des moustiques An. gambiae, Ae. albopictus et Cx. quinquefasciatusMosquitoes are the main vectors involved in the transmission of pathogens to humans. Accurate identification of mosquito species is crucial to distinguish between vector and non-vector species. The mosquito blood meal determination is fundamental in understanding the behavior of vector species. Thus, we have listed 106 mosquito species currently recorded in Mali, including 28 Anophelinae and 78 Culicinae. Then, we evaluated the effectiveness of MALDI-TOF MS for identified mosquitoes collected in Mali and to determine their blood meal source. The results obtained show the ability of MALDI-TOF MS to identify mosquitoes collected in Mali and their source of blood meal. Subsequently, we were able to confirm the robustness of MALDI-TOF MS to identify other animal blood samples. We artificially engorged Anopheles gambiae and Anopheles coluzzii on eight animal bloods samples. We obtained 100% correct identification of the blood source for samples taken 1 to 24 hours after feeding. Then, we experimentally engorged An. gambiae, An. coluzzii and Ae. albopictus on successive and mixed blood meals using MALDI-TOF MS. The results revealed that MALDI-TOF MS is able to identify mixed blood meals. In addition we used MALDI-TOF and culturomics for the microbiota study of the mosquito collected in the field, notably in Marseille and Mali. The culturomics approach revealed a great diversity of the digestive microbiota of the An. gambiae, Ae. albopictus and Cx. quinquefasciatus mosquitoes
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Krüger, Ralf. "Untersuchungen zum Einbau von Analytionen in MALDI-Matrizes sowie zur Ionisation und Adduktbildung in der MALDI-Massenspektrometrie." [S.l. : s.n.], 2003. http://deposit.ddb.de/cgi-bin/dokserv?idn=969681682.
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Kirmess, Kristopher Michael. "Investigation of Primary Ion Formation Mechanisms in UV-MALDI-MS Using Excited State Dynamics of Common MALDI Matrices." OpenSIUC, 2015. https://opensiuc.lib.siu.edu/dissertations/1110.
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The motivation of this dissertation is to provide insight towards primary ionization mechanisms within MALDI mass spectrometry. Albeit MALDI-MS is an extensively used analytical technique, the mechanism in which primary ions are created is still under scrutiny. Two current models of primary ionization exist which claim to elucidate the ion formation mechanisms within MALDI. In this work, excited state dynamics of MALDI matrices are shown to play an important role in the ionization mechanism. Upon inspection of the thermodynamic properties of commonly used MALDI matrices, no correlation was observed when plotted against their respective analyte ion yields. However, the excited state singlet lifetimes of these matrices seem to correlate well with their respective analyte ion yields. In the broadest sense, this correlation further supports the fact that photophysical properties of the matrix should be included in current UV-MALDI models. Investigation of a claim which stated singlet energy pooling reactions were absent in the MALDI matrix 2,4,6-trihydroxyacetophenone (THAP) resulted in the discovery of a new energy pooling mechanisms. Characteristic of aromatic ketones such as THAP, intersystem crossing is an efficient process in solution, which gives way to fluorescence in the solid state. Triplet pooling mechanisms from two neighboring THAP molecules are proposed and appear to be dependent on the preparation solvent used. These triplet pooling reactions are thought to play an important role in the primary ion formation mechanism within MALDI. To further investigate the theory of triplet species playing a vital role in MALDI ionization, the internal heavy-atom effect was employed to determine the effect of the triplet species. MALDI mass spectra and excited state decays of these heavy-atom substituted matrices were collected to demonstrate the relationship between triplet species and analyte ionization efficiency. Gas-phase thermodynamics and absorption at 337 nm were also examined to determine if these properties affected the analyte ion signal observed in the MALDI mass spectrum. Using the information collected from the previous study, an advanced MALDI matrix is synthesized. Addition of covalently bound iodine to the gold standard matrix, α-cyano-hydroxycinnamic acid, should drastically improve the performance of the non-substituted matrix due to the increase in triplet species present for pooling reactions. Sample preparation methods in MALDI are examined as are the effects of crystal morphology on the overall signal observed in the mass spectrum. Exciton hopping and pooling rates are highly dependent on intermolecular interactions, so it is expected that crystal packing will affect MALDI. As noted for THAP, preparation solvent plays a significant role in not only crystal morphology, but also the excited state dynamics for all matrices studied.
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Collazo, Verena. "Reverse Sanger-Sequenzierung mittels MALDI-TOF-Massenspektrometrie." [S.l.] : [s.n.], 2001. http://deposit.ddb.de/cgi-bin/dokserv?idn=964226871.
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Enebro, Jonas. "Characterization of carboxymethyl cellulose by MALDI-TOFMS /." Stockholm : [Fiber och polymerteknologi, Kungliga Tekniska högskolan], 2007. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-4376.
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Leander, Ellinor. "Artidentifiering av mögelsvamp med MALDI-TOF MS." Thesis, Linnéuniversitetet, Institutionen för kemi och biomedicin (KOB), 2018. http://urn.kb.se/resolve?urn=urn:nbn:se:lnu:diva-80166.
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Snabb och korrekt artidentifiering är avgörande för effektiv behandling av svampinfektioner, särskilt bland immunsupprimerade patienter. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) används rutinmässigt på kliniska laboratorier för identifiering av karaktäristiska proteinmönster hos bakterier och jästsvampar genom tolkning av proteinspektra i en masspektradatabas för korrekt artidentifiering. Mögelsvamparnas hårda cellvägg och heterogena växtsätt med varierande proteinuttryck beroende på mognadsstadie, försvårar identifiering med MALDI-TOF MS. Metodens tänkbara fördelar mot traditionella metoden mikroskopering är förkortade svarstider, säkrare artidentifiering av fler arter och mindre beroende av subjektiv morfologisk bedömning. Studiens syfte var att undersöka om MALDI-TOF MS kunde anpassas och användas för identifieringen av mögelsvamp i klinisk rutindiagnostik. Fyra referensstammar (Aspergillus niger, A. fumigatus, A.terreus, A.flavus) och ett kliniskt isolat (A.terreus) undersöktes. Preparationsmetoderna (I) fullständig myrsyraextraktion, (II) direktapplicering och (III) suspension i destillerat vatten användes för analys av sporer och frontmycel hos yngre och äldre mögelkulturer. Två olika masspektradatabaser för artidentifiering jämfördes; rutindatabasen BDAL och den specialiserade mögeldatabasen Filamentous Fungi Library. Även plocktekniken av mögelmaterial inför analys med MALDI-TOF MS utvärderades. Vid vissa tillfällen förbättrades artidentifieringen efter extraktion av mögelkulturerna, medan i andra fall var direktapplicering fullt tillräcklig. Mögelmaterial med mycket sporer tenderade ge något fler artidentifieringar i BDAL oavsett kulturernas ålder. Filamentous Fungi Library tenderade i vissa fall ge bättre resultat jämfört med BDAL för yngre kulturer. Fler studier krävs för att utvärdera och optimera MALDI-TOF MS som metod för artidentifiering av mögelsvamp.Rapid and accurate species identification is crucial for successful treatment of fungal infections, especially among immunosuppressed patients. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is used routinely at clinical laboratories to identify characteristic protein patterns of bacteria and yeast by the interpretation of protein spectra in a database for accurate species identification. The hard cell wall of the mold and the heterogeneous growth with varying protein expression due to maturation, complicates identification with MALDI-TOF MS. The potential benefits of this method compared to microscopy as traditional method are shortened turn-around times, safer species identification of more species that is independent on subjective morphological assessment. The purpose of the study was to investigate whether MALDI-TOF MS could be adapted and used for the identification of molds in clinical routine diagnostics. Four reference strains (Aspergillus niger, A.fumigatus, A.terreus, A.flavus) and a clinical isolate (A.terreus) were examined. The preparation methods (I) complete formic acid extraction, (II) direct application and (III) suspension in distilled water were used for analysis of spores and frontmycelium from younger and older mold cultures. Two different masspektradatabases for species identification were compared; routine database BDAL and the specialized mold database, Filamentous Fungi Library. Also the collecting technique of mold prior to analysis with MALDI-TOF MS was evaluated. Sometimes, the species identification improved after extraction of mold cultures, while in other cases direct application was sufficient. Cultures with a lot of spores tended to give slightly more species identifications in BDAL regardless of the age of cultures. Filamentous Fungi Library, in some cases, tended to improve the performance compared to BDAL for younger cultures. More studies are required to evaluate and optimize MALDI-TOF MS as a method of mold identification.
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Stauber, Jonathan. "Imagerie MALDI : nouveaux développements et applications cliniques." Lille 1, 2007. https://pepite-depot.univ-lille.fr/LIBRE/Th_Num/2007/50376-2007-379.pdf.
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Les avancées de la biologie moléculaire se sont également réalisées avec l’évolution des techniques d’imagerie dans les domaines de la génomique. La transcriptomique et plus récemment de la protéomique grâce ù l'essor d'un outil essentiel, la spectrométrie de masse. Cette technique a trouvé sa place pour générer des profils protéiques caractéristiques de l'état physiologique de la cellule, de fluides complexes tels que le sang ou les urines. Elle apparaît aujourd'hui comme un outil indissociable de la recherche en biologie et en médecine. Les nouveaux développements tendent à conduire la spectrométrie de masse vers l’imagerie moléculaire pour l'identification de pathologies, pour la distribution de médicament au sein d’un animal. Ou pour une utilisation en diagnostique et en pronostique. Cette technologie récente, demande qu'à être développée, améliorée, standardisée. C’est dans ce cadre que ma thèse intitulée « Imagerie MALDI nouveaux développements et applications cliniques » a été orientée. Les différents résultats ont permis de développer I’imagerie spécifique moléculaire MALDI, I’imagerie moléculaire MALDI de tissus fixés et paraffinés avec des applications à la fois dans le cadre de la maladie de Parkinson et le cancer de l’ovaire. L’évolution en filagramme de cette technologie d'imagerie moléculaire semble devoir se réaliser de paire avec les techniques d’imagerie non invasive pour devenir une nouvelle technologie en cliniqueThe recent innovations in molecular biology were realized with the evolution of the imaging techniques in the field of Genomics, Transcriptomics, and recently in Proteomics with an essential tool, the mass spectrometry. This imaging technique create characteristic protein profiles of the cellular states, and appears today as an undissociable tool for research in biology and medicine. The last developments look to emerge the mass spectrometry to a molecular imaging to identify pathologies, to observe the drugs distributions in tissues, or the diseases diagnosis or prognosis. This unique and recent technology should be developed, improved, and standardized. It's in this point of view the my PhD training named MALDI imaging new developments and clinical applications was defined. The different results obtained during my PhD were permits to create a concept of Specific Imaging Mass Spectrometry, to develop Molecular MALDI imaging of frozen and FFPE tissues with many applications in the research of specific biomarkers in Parkinson disease and ovarian cancer. The evolution of this unique molecular imaging technique should be in the next years a complementary method of others in vivo imaging technique
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Jacksén, Johan. "Improved techniques for CE-MALDI-MS off-line coupling and MALDI-MS analysis of primarily hydrophobic proteins and peptides." Licentiate thesis, KTH, Chemistry, 2007. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-4599.
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Due to the hydrophobic nature of integral membrane proteins (IMP) they give rise to several difficulties concerning handling and analysis, which is not the case for the most water soluble proteins. New analysis methods are needed, where the insolubility problems of the hydrophobic proteins due to aggregation and adhesion are tackled. Those problems also affect digestion performance and equipment compatibility for the analysis.
Protocols for analysis and separation specified for IMP are presented in Paper I and III.
The instrumentation used in this work was capillary electrophoresis (CE) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Both instruments are suitable for peptide/proteins analysis.
In Paper I, protocols for a CE separation of bacteriorhodopsin (BR) peptides as model IMP peptides are established. Also, a partially automated manufacturing procedure of a concentration MALDI-target is presented, suitable for fractions from CE. The MS analysis detected 9 out of 10 cyanogen bromide (CNBr) digested BR peptides. A novel technique for the off-line integration of CE to MALDI-MS using a closed-open-closed system is presented in Paper II, where the open part is a microcanal functioning as a MALDI target window. Investigation of the microcanal electro-osmotic flow (EOF) properties and band broadening characteristics was performed. A protein separation was obtained and detected with MALDI-MS analysis in the microcanal. Different protein digestion methods were evaluated using BR in Paper III through MALDI-MS. Several digestion methods as well as MS media were investigated alongside different MALDI matrices. For example, matrices as the hydrophobic 2,6-dihydroxyacetophenone (DHAP) and 2-Hydroxy-3-methoxybenzoic acid (2H3MBA) or 2-Hydroxy-5-methoxybenzoic acid (2H5MBA) mixed with DHB, appeared to be promising matrices for analysis of BR.
Med anledning av integrala membranproteiners (IMP) hydrofoba egenskaper uppstår flera svårigheter vid hantering och analys av IMP, vilket inte är fallet för vattenlösliga proteiner. Nya analysmetoder krävs, som löser löslighetsproblemen för de hydrofoba proteinerna som tex flockning och adsorbtion. Dessa problem påverkar även klyvningsgrad och kompatibilitet med analysutrustningen.
I Artikel I och Artikel III presenteras protokoll för analys och separation specifikt för IMP. Instrumenteringen som har använts i detta arbete är kapillärelektrofores (CE) och matris-assisterad laserdesorptions-joniserings-masspektrometri (MALDI-MS). Båda instrumenten är lämpade för peptid/protein analyser.
I Artikel I, presenteras protokoll för en CE separation av peptider från bacteriorhodopsin (BR), som användes som modellpeptider för IMP. En delvis automatiserat tillverkningsprocedur för en koncentrerande MALDI-platta, som är anpassad för CE fraktionerna beskrivs också. MS-analysen detekterade 9 av 10 BR-peptider från cyanobromid-klyvning (CNBr). En ny teknik för off line-integrering av CE till MALDI-MS genom ett slutet-öppet-slutet system presenteras i Artikel II, där den öppna delen är en mikrokanal som fungerar som detektionsfönster i MALDI. Undersökning av mikrokanalens egenskaper som tex det elektroosmotiska flödet (EOF) och bandbreddningen utvärderades. En proteinseparation genomfördes och detekterades med MALDI–MS i mikrokanalen. Olika proteinklyvningsmetoder för BR undersöktes i Artikel III med MALDI-MS. Flera proteinklyvningsmetoder samt MS-medier utvärderades tillsammans med olika MALDI-matriser. Den hydrofoba matrisen 2,6-dihydroxyacetophenone (DHAP) och 2-Hydroxy-3-methoxybenzoic acid (2H3MBA) eller 2-Hydroxy-5-methoxybenzoic acid (2H5MBA) blandade med DHB, visade sig exempelvis vara lovande matriser för BR-analyser.
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Neubert, Hendrik. "An investigation into the enhancement of MALDI-MS by modification of surface chemistries (MALDI, matix-assisted laser desorption/ionisation)." Thesis, King's College London (University of London), 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.396347.
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PENG, LIJUAN. "MATRIX-ASSISTED LASER DESORPTION/IONIZATION (MALDI) TARGET MODIFICATION FOR ENHANCED PROTEOMICS ANALYSIS AND PLASMA POLYMER CHARACTERIZATION BY MALDI MASS SPECTROMETRY." OpenSIUC, 2010. https://opensiuc.lib.siu.edu/dissertations/207.
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The work described in this dissertation is divided into three sections. In the first section three surface modifications are used to produce MALDI targets having reduced surface-protein binding affinity with a goal of increasing peptide/protein MALDI ion signals and lowering the limits of detection (LODs) for proteins and peptides. The second section discusses a bioselective MALDI target, produced via radio frequency (rf) plasma deposited ethylenediamine (EDA), for on-target separation of complex protein mixtures. The third section develops a new approach for characterization of rf plasma-deposited bulk polymers by using MALDI MS. Previous studies in our group have shown that the analyte signal in a MALDI MS experiment is strongly influenced by the binding interactions between the target surface and the analyte. Specifically, the analyte signal increases with decreasing surface-analyte binding affinity, which has been attributed to more unbound analyte being available for incorporation within the MALDI matrix. In the presented studies MALDI targets are modified with polyethylene glycol (PEG)-like structures via chemical grafting of PEG onto polyurethane (PU) film and rf plasma polymerization of ethylene oxide vinyl ether (EO2) and tetraglyme. It is shown that there are enhancements in the protein MALDI ion signals on these modified targets and that the LOD for target proteins is decreased by a factor of 2-10 in comparison with the conventional stainless steel MALDI target. On-probe affinity capture (OPAC) MALDI MS, developed in our group, has shown that functional group modified MALDI targets can be used to rapidly and selectively isolate target analytes from complex samples. For applications involving analysis of complex peptide/protein mixtures, fractionation of the mixture on the basis of component pI can reduce MALDI ion suppression effects leading to efficient ionization of larger numbers of mixture components. In the present studies a MALDI target is modified by rf plasma deposition of polymerized EDA to yield an OPAC target suitable for capture of proteins with low pI (expected to be negatively charged at neutral pH). In subsequent MALDI MS analyses of both control and biological mixtures after fractionation on the OPAC target it is observed that a significant number of additional peptide/protein ion signals are detected. The results of these studies, along with studies of the effects of the density of the primary amine functionality on the bio-selective MALDI ion signals, are presented. The complex nature of the polymer films resulting from plasma polymerization makes it very difficult to characterize their molecular structures. The presented study is the first to use MALDI MS for characterization of rf plasma-deposited bulk polymers and for investigation of the rf plasma polymerization process. It is shown that the mass spectra of the soluble fraction of allyl alcohol, EO2 and ethylene glycol butyl vinyl ether -plasma polymers contain clear polymer series. Furthermore, it is found that the peaks of the EO2-plasma polymer series shift to higher molecular weight distribution with decreasing plasma duty cycle. In contrast to predictions based on conventional radical polymerization, the mass spectra of all three plasma polymers exhibit the same repeat unit of 44 Da, for which the most likely structure would be -(CH2CH2O)-.
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Dallacker-Losensky, Kevin. "Identifizierung von obligaten Anaerobiern der Bacteroides fragilis Gruppe einschließlich Metronidazol-resistenter und Enterotoxin-positiver Stämme mittels MALDI-TOF MS." Doctoral thesis, Universitätsbibliothek Leipzig, 2016. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-206555.
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Die klassische Identifizierung von obligat anaeroben Bakterien ist mit einem hohen Labor- und Zeitaufwand verbunden. Um festzustellen, ob die Identifizierung mittels Matrix-unterstützter Laser-Desorption/Ionisation und Massenspektrometrie mit Flugzeitanalysator (Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry; MALDI-TOF MS) ein Verfahren ist, um obligate Anaerobier eindeutig zu identifizieren, wurde mit der vorliegenden Arbeit die Identifizierung von unterschiedlichen Spezies der B. fragilis Gruppe mittels MALDI-TOF MS untersucht.
Hierfür wurden 105 obligate Anaerobier der B. fragilis Gruppe aus der Stammsammlung des Institutes für Medizinische Mikrobiologie und Infektionsepidemiologie der Universität Leipzig untersucht. Es fanden sich für die untersuchten Erreger Spektren mit sehr guter Auflösung. Eine Identifizierung und Differenzierung war eindeutig möglich. Unter Verwendung dieser Daten wurde eine Referenzdatenbank erstellt. Die erhaltenen Ergebnisse wurden mittels einer verblindeten Studie überprüft, wobei 52 von 53 (98,1%) der untersuchten Stämme eindeutig identifiziert werden konnten. Dies schließt ebenfalls die Identifizierung und Differenzierung von 15 Metronidazol-sensiblen/ Enterotoxin-negativen, 8 Metronidazol-resistenten/ Enterotoxin-negativen und 8 Metronidazol-sensiblen/ Enterotoxin-positiven B. fragilis Stämmen ein.
Die Identifizierung mittels MALDI-TOF MS ist somit eine zuverlässige Methode zur Identifizierung von obligaten Anaerobiern der B. fragilis Gruppe. Weiterhin finden sich Hinweise, dass ein Nachweis von Resistenz-, Virulenz- und Pathogenitätsfaktoren mittels MALDI-TOF MS bei diesen Erregern möglich ist.
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Zechmann, Carsten. "Spektroskopische Untersuchung der Lumineszenzerscheinungen beim UV-MALDI-Desorptionsprozess." [S.l. : s.n.], 2001. http://e-diss.uni-kiel.de/diss/d440.pdf.
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Bouschen, Werner. "Ortsaufgelöste MALDI-Massenspektrometrie an biologischen und synthetischen Oberflächen." [S.l. : s.n.], 2004. http://deposit.ddb.de/cgi-bin/dokserv?idn=971766886.
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Kempka, Martin. "Improved mass accuracy in MALDI-TOF-MS analysis." Licentiate thesis, Stockholm : Division of Analytical Chemistry, Royal Institute of Technology, 2005. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-313.
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Kriegsmann, Mark. "MALDI MS Imaging zur Untersuchung von synovialem Gewebe." Doctoral thesis, Universitätsbibliothek Leipzig, 2013. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-118897.
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Enthaler, Bernd [Verfasser]. "MALDI imaging in ex-vivo skin / Bernd Enthaler." München : Verlag Dr. Hut, 2014. http://d-nb.info/1049363086/34.
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Halgunset, Anders. "Typing av Legionella pneumophila med MALDI-TOF MS." Thesis, Norges teknisk-naturvitenskapelige universitet, Institutt for bioteknologi, 2012. http://urn.kb.se/resolve?urn=urn:nbn:no:ntnu:diva-24697.
Full textAbstract:
Legionella pneumophila er fakultativ intracellulære bakterier som lever naturlig i akvatiske habitater. De kan replikere i makrofager og protozoer, bli overført til mennesker gjennom areosoler og føre til smitte. Ved eventuelle utbrudd er det viktig å kunne identifisere kilden, noe som i dag bli gjort ved å typebestemme L. pneumophila med sekvensbasert typing (SBT). Samtidig har matrix-assisted laser desorption/ionization ? time-of-flight (MALDI?TOF) mass spectrometry (MS) vist seg å kunne brukes til rask og effektiv identifisering av en rekke mikroorganismer på artsnivå, deriblant Legionella. Hos enkelte andre mikroorganismer har MALDI?TOF MS også vist å kunne skille mellom underarter. MALDI?TOF MS ble i denne oppgaven benyttet for å undersøke om det var mulig å skille L. pneumophila fra hverandre på stammenivå ved hjelp av MALDI Biotyper 3.0 programvare. Standardprotokollen anbefalt av Bruker Daltonics for proteinekstraksjon fra mikroorganismer ble tilpasset/optimalisert for bruk på L. pneumophila, slik at reproduserbare massespektre ble oppnådd. Evalueringen viste at proteinekstraksjon fra ca 2 µl biologisk materiale med 10 µl acetonnitrill og 10 µl maursyre, ga best scoreverdi opp mot Bruker Daltonics referansebibliotek. Den optimale temperaturen og varigheten for dyrkning ble vist å være henholdsvis 37 ˚C og 48 ± 2 timer. Det ble vist at lagring av skåler/kolonier med L. pneumophila ved 37 ˚C førte til forandringer i massespektrene, mens lagring ved 20 ˚C ikke medførte slike forandringer og at stabile massespektre ble oppnådd etter seks dagers lagring.Det ble bygget opp et referansebibliotek i MALDI Biotyper 3.0 bestående av referansespekter/referansetopplister (MSP) fra til sammen 48 Legionella spp. hvorav 41 var L. pneumophila -isolater. Med referansebiblioteket ble det vist at MSP (dannet ved standard innstillinger) for mange L. pneumophila var for like hverandre til å gi identifisering mot egne MSP. En klyngeanalyse ble brukt i sammenligning av L. pneumophila mellom SBT og MSP fra referansebiblioteket dannet med MALDI?TOF MS. Sammenligningen viste at diversiteten i proteinsammensetningen hos L. pneumophila, representert som MSP, ikke ga samme oppløselighet som gjennom sekvensvariasjonene fra genene i SBT -analysen.
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Trim, Paul James. "MALDI-MS imaging for direct drug distribution analysis." Thesis, Sheffield Hallam University, 2009. http://shura.shu.ac.uk/20455/.
Full textAbstract:
MALDI Imaging has gained huge interest in the past few years with an ever increasing population of specialists choosing to investigate samples using MALDI imaging, including growing interest and financial backing from pharma and contract research organisations. Presented within this thesis is the development and application of MALDI imaging techniques for a variety of analytical problems. The use of various software packages have been employed in the interpretation of the data acquired from MALDI experiments including, the use of statistical analysis for the identification of ion of interest from 6 distinct brain regions and also for the identification of ions of interest associated with small molecule tumour markers. The advantages of MALDI-IMS-MSI as a further separation stage within MALDI-MSI have been shown. Demonstrated is a method for MALDI-IMS-MS imaging of endogenous lipids in healthy tissue and tumours, also demonstrated is the application of MALDI-IMS-MS to xenobiotic distribution studies, it has been clearly shown that ion mobility separation within MALDI-MSI experiments can improve the analysis of xenobiotics by removing any interfering ions. With instrumentation development for MALDI a high repetition rate Nd:YVO4 laser has been assessed as a possible method for decreasing acquisition time.
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Sun, Zhen. "Studies of Atmospheric Pressure Visible-Wavelength MALDI-MS." University of Toledo / OhioLINK, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=toledo1333747638.
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Eastwood, Stephanie. "Development of Amine Containing Polymer Modified MALDI Targets for Complex Peptide and Protein Mixture Fractionation Prior to MALDI Mass Spectrometry Analysis." OpenSIUC, 2014. https://opensiuc.lib.siu.edu/dissertations/948.
Full textAbstract:
AN ABSTRACT OF THE DISSERATION OF STEPHANIE EASTWOOD, for the Doctor of Philosophy degree in CHEMISTRY, presented on the DATE OF DEFENSE, at Southern Illinois University Carbondale. TITLE: DEVELOPMENT OF AMINE CONTAINING POLYMERIC MODIFIED MALDI TARGET SUBSTRATES FOR COMPLEX MIXTURE FRACTIONATION OF PEPTIDE AND PROTEIN MIXTURES PRIOR TO MALDI MASS SPECTROMETRY ANALYSIS MAJOR PROFESSOR: Dr. Gary Kinsel The focus of this dissertation is the synthesis, development and application of the amine containing modified targets for the fractionation of complex mixtures of peptides and digested proteins prior to MALDI-MS analysis. Even though MALDI-MS is increasingly used in proteomic applications and analysis, this method suffers from loss of performance in the analysis of mixtures, due to the ion suppression effect. In this effect, basic peptides ionize more readily than acidic peptides and there by suppress the ionization and detection of less basic peptides resulting in the loss in the detection of valuable sequence information and posttranslational modifications. In the approach taken, a modified target containing amine functionality was utilized as a mixture fractionation substrate. The substrates were chosen for the adsorption of targeted analytes. Incorporating this amine function into 3D brushes offer the ability to absorb higher quantities of peptides, allowing for a more thorough analysis and more sequence coverage. In this work fractionation occurs as the result of electrostatic attraction or repulsion between the positively charged polymer surface and negatively charged analytes in solution. Fractionation studies utilizing this amine chemistry for separation were done first with binary peptide mixtures. Basic peptides are observed to repel from the cationic polymer surfaces, allowing the suppressed acidic peptides to separate from the mixture by binding to the amine substrates. The acidic peptides can be eluted and observed. Following successful demonstrations, more complex mixtures of peptides derived from enzymatic tryptic digestion of proteins were studied. Following fractionation, MALDI mass spectra of both the washed and eluted fractions are obtained and ions signals are analyzed and associated with predicted peptide sequences. Complete analysis of the ion signals using Prowl and PeptideMap leads to a percent protein sequence coverage. Comparison of this number with the percent sequence obtained from unfractionated (conventional) digested peptide mixture mass spectra reveals that fractionation does in fact significantly increase and also increases discovery of post translational modifications (PTM). Fractionation studies were studied by varying the pH of tryptically digest proteins. Sequence coverages were found to increase upon fractionation at all pHs studied. In order to increase the discovery of PTMs, the cationic polymer brush was doped with copper ions to selectively capture phosphopeptides. Negatively charged phosphopeptides were shown to be selectively captured by the copper ions complexes within the polymer brush substrate, and subsequently released upon addition of acid.
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Seyer, Alexandre. "Imagerie par spectrométrie de masse : développements méthodologiques et applications biologiques." Thesis, Evry-Val d'Essonne, 2010. http://www.theses.fr/2010EVRY0028/document.
Full textAbstract:
Mon travail de thèse a consisté à poursuivre le développement de l’Imagerie par Spectrométrie de Masse (IMS), à la fois sur le plan méthodologique mais aussi à travers des applications biologiques.Dans une première partie est exposé le développement d’une méthode de préparation des échantillons de très petite taille pour l’imagerie chimique, et plus particulièrement pour l’imagerie TOF-SIMS. Cette méthode a pu être validée en étudiant des molécules de la famille des flavonoïdes dans différents types de graines d’Arabidopsis thaliana ne mesurant que 400 nm de diamètre. La seconde partie, dédiée aux applications biologiques, est divisée en deux sections. La première section regroupe deux sujets où il était question de détecter et de localiser, à l’aide de l’imagerie TOF-SIMS, la molécule active d’une crème anti-acné dans des coupes de peaux de cadavre humain, ainsi qu’un retardateur de flamme bromé, le décabromodiphényl éther, dans ses tissus cibles chez le rat. Dans la seconde section, nous avons étudié, en imagerie TOF-SIMS et MALDI-TOF, l’absorption des lipides durant la digestion et enfin, avec l’aide d’outils d’analyse statistiques, nous avons comparé les profils lipidiques d’échantillons sains et atteins de la mucoviscidose chez un animal modèle de la maladie.À travers ces différents projets, nous avons pu conclure que les imageries par spectrométrie de masse TOF-SIMS et MALDI-TOF sont deux techniques complémentaires et qui, combinées à l’analyse statistique, peuvent être de puissants outilsMy PhD’s work consisted in continuing the development of Mass Spectrometry Imaging (MSI) methods, in terms of methodology improvements but also through biological applications.The first part concerned the development of a novel sample preparation method dedicated to very small objects for chemical imaging, particularly for TOF-SIMS imaging. This method has been validated by studying different types of flavonoids in from seeds of Arabidopsis thaliana, with a size of 400 nm only. The second part, dedicated to biological applications, is divided into two sections. The first section includes two projects where the goals was to detect and locate, using TOF-SIMS imaging, the active molecule of an anti-acne cream in human skin sections, and a brominated flame retardant, the decabromodiphenyl ether, in target tissues in rats. In the second section, we have studied by MALDI-TOF and TOF-SIMS imaging the lipid absorption during the digestion, and finally, with the help of statistical analysis tools, we compared lipid profiles of healthy samples versus those from cystic fibrosis samples in a model animal of the disease.Through these projects, we have concluded that MALDI-TOF and TOF-SIMS imaging are two complementary techniques, and, when they are combined with statistical analysis, they can be powerful tools
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Sandonato, Beatriz Brabetz [UNESP]. "Peptídeos cíclicos hepatotóxicos: MALDI-TOF como uma ferramenta analítica." Universidade Estadual Paulista (UNESP), 2014. http://hdl.handle.net/11449/123921.
Full textAbstract:
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000832493.pdf: 1769807 bytes, checksum: e6c5bb7d9bcec2a173427a67060a8dce (MD5)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
As cianobactérias são bactérias fotossintetizantes que podem ser encontradas nos mais variados ambientes. Algumas espécies de cianobactérias produzem toxinas denominadas cianotoxinas. As cianotoxinas denominadas microcistinas (MCs) são heptapeptídeos cíclicos hepatotóxicos produzidas durante florações de cianobactérias. Sua detecção e quantificação em mananciais são de grande importância devido aos danos a saúde humana que essas microcistinas podem causar. Atualmente, a técnica MALDI-TOF-MS tem se mostrado uma eficiente ferramenta para a análise de microcistinas, e recentes estudos tem reportado seu uso na quantificação destas. Diante do exposto, o objetivo desta dissertação foi a avaliação de diferentes matrizes e diferentes métodos de preparo de amostra para MALDI-MS visando a detecção e quantificação das microcistinas. Com a finalidade de alcançar o melhor método de quantificação das microcistinas, nove matrizes para MALDI, onze métodos de preparo de amostra mais um método que foi adaptado por nosso grupo de pesquisa, o método Vacuum drying adaptado, e dois padrões comerciais de microcistinas, MC-LR e MC-RR, foram utilizados. A avaliação dos métodos de preparo de amostra foi realizada utilizando como matriz o ácido α-ciano-4-hidroxicinâmico (HCCA) e o peptídeo angiotensina I como padrão interno. Os métodos foram avaliados com relação aos seus coeficientes de variação (CV), suas cristalizações e espectros obtidos, utilizando a MC-RR como amostra. O método Vacuum drying adaptado apresentou os melhores resultados e sua curva analítica foi construída utilizando ambas as variantes de microcistinas. O limite de detecção (MLD) e o limite de quantificação (LDQ) também foram calculados. Cada curva da variante de microcistina apresentou excelente linearidade e os valores de r variaram entre 0,98-99, demonstrando que o método é adequado para a quantificação destas. Após as análises dos...
Cyanobacteria are photosynthetic bacteria that can be found in diverse environments. Some species of cyanobacteria produce toxins denominated cyanotoxins. The cyanotoxins called microcystins (MCs) are hepatotoxic cyclic heptapeptides produced during cyanobacterial blooms. Its detection and quantification in springs are of great importance due to damage to human health that these microcystins can cause. Currently, MALDI-TOF-MS technique has been shown to be an efficient tool for the analysis of microcystins, and recent studies have reported its use in the quantification of these. On the exposed, the aim of this thesis was to evaluate the different matrices and different methods of sample preparation for MALDI-MS aimed at the detection and quantification of microcystins. With the aim of achieving the best method of quantification of microcystins nine matrices for MALDI eleven methods of sample preparation over a method that was adapted by our research group, the adapted vacuum drying method, and two commercial standards of microcystins, MC-LR, and MC-RR, are used. The evaluation methods of sample preparation was performed using as matrix the α- cyano-4-hydroxycinnamic acid (CHCA) and the angiotensin I as internal standard. The methods were evaluated with respect to their coefficients of variation (CV), their crystallization and spectra obtained using the MC-RR as a sample. The adapted vacuum drying method showed the best results and their analytical curve was constructed using both variants of microcystins. The limit of detection (MDL) and the limit of quantification (LOQ) were also calculated. Each curve variant of microcystin showed excellent linearity and r values ranged from 0.98 to 99, showing that the method is suitable for quantifying these. After analysis of the methods of sample preparation, the nine matrices were evaluated with respect to CV, crystallization and spectra, using the MC-RR as a sample. The best results were obtained...
FAPESP: 2012/03663-8
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uk, siricordcc@yahoo co, and Cornelia Charito Siricord. "Detection of Phytophthora species by MALDI-TOF mass spectrometry." Murdoch University, 2005. http://wwwlib.murdoch.edu.au/adt/browse/view/adt-MU20070717.125452.
Full textAbstract:
Phytophthora diseases have caused worldwide economic, social and environmental
impacts for decades. Once their presence is confirmed, they are difficult to eradicate. To
reduce and manage the damage inflicted by the pathogen, fast and reliable disease
management protocols are required. Tests that enable the rapid and reliable
identification of the pathogen assist greatly in disease management.
Phytophthora species are traditionally not only detected by baiting but also by plating of
symptomatic tissue on selective media. Species can be identified by the characteristics
of the mycelium growing out of the bait. However, the method is low throughput,
labour intensive, and prone to false negatives. An alternative approach would be to
detect the pathogen by the presence of its DNA. This involves amplification of the
pathogen DNA using Polymerase Chain Reaction (PCR) and detection of the
amplification product. Detection is usually by agarose gel electrophoresis. However,
this is also a labour intensive process involving pouring, loading, running, and staining
of the gels. The aim of this thesis is to explore the use of Matrix Assisted Laser
Desorption/ Ionisation Time-of-Flight (MALDI-TOF) mass spectrometry for detection
of PCR products. This procedure enables the analysis of large numbers of samples
within a very short time-frame as the average time for analysis of each sample is in the
order of milliseconds.
The assay involves annealing an extension (genotyping) primer to the PCR product and
its extension by a single nucleotide. The nature of the nucleotide added differentiates
species as does the site to which the primer anneals. Multiple extension (genotyping)
primers can be used together in a single reaction for detection of multiple species. In
this project four genotyping primers (GPs) were designed from the ITS regions of
Phytophthora palmivora, Phytophthora cinnamomi, Phytophthora citricola, and
Phytophthora cambivora.
The extension primers were tested for their specificity on the DNA of the target species.
The four primers designed were specific for their intended targets except for GPpalm3
which in addition to being extended by ddT when tested with DNA from P. palmivora,
was also extended by ddC when tested with DNA from other species of Phytophthora
or Pythium.
These primers were also tested for their ability to detect multiple Phytophthora species
in a single reaction (multiplexing). Mixtures of primers were added to mixed DNA
templates and the primer extension reaction carried out. The primers were designed so
that their masses were sufficiently different for them to be identified from a mixture.
Six replicates were analysed for each reaction. In general only about 1-3 of the six
replicates gave a positive reaction. This indicates that there may be some interference
between primers, or that the presence of all four nucleotides interfered with the primer
extension reaction. Increasing either the amount of enzyme, the amount of nucleotides
or both did not improve the results.
The sensitivity of detection was tested by the addition of different amounts of mycelium
to soil. The detection sensitivity depended on the primer pair used for PCR
amplification. The ITS1/2 primer pair was more sensitive than the ITS1/4 pair. The
limit of detection was 1 ìg mycelium g soil-1. However using nested PCR, levels of
sensitivity comparable to those obtained using the ITS1/2 primer pair could be
achieved. Primers to other regions of the genome such as the beta cinnamomin elicitin
gene gave very low levels of sensitivity compared to the ITS primers.
In comparison with DNA detection we found that the limit of detection using baiting
was 4 ìg mycelium g soil-1. Results below this limit were unreliable. The method
suffered from the additional disadvantage that it took a long time in comparison to DNA
detection.
DNA detection methods do not distinguish between living and dead organisms in the
soil. However it can be hypothesised that DNA is unlikely to persist for any significant
length of time in soil. To test this, we added plasmid DNA to soil and tested the
persistence of this DNA using a variety of methods such as precipitation of labelled
DNA, southern blotting and PCR amplification. It was found that in general, in soils
from different ecosystems, the bulk of the DNA was undetectable after 24 hours. The
rate of DNA breakdown differed with the soil type. In some soils, the added DNA was
not detected even after 2 hours, whereas in others it could be observed after 10 hours.
The detection depended on the method. Southern blotting showed that although DNA
could be observed at 10 hours, by 24 hours it was completely degraded. In contrast a
PCR product could be obtained from the soil extracts up to 24 hours. In a separate
experiment, plasmid DNA was detectable over a 24 hour incubation period in 5 soil
samples from 5 different sites. The results suggest that DNA is degraded rapidly in soil
and is unlikely to persist longer than 24 hours.
The results in this thesis demonstrate that MALDI-TOF MS is a suitable alternative to
agarose gel electrophoresis for analysis of PCR products. The technique is rapid,
differentiates species from mixtures, is high-throughput and amenable to automation.
Implementation will require further research to automate the primer extension assay to
reduce the sensitivity to impurities in the DNA and to design parameters for sampling
asymptomatic material.
26
Bertilsson, Sarah. "Glycanmapping of glycoproteins with UPLC-FLR-MALDI/TOF-MS." Thesis, Uppsala universitet, Analytisk kemi, 2014. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-228040.
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Wyatt, Mark Francis. "Analysis of acrylic polymers by MALDI-TOF mass spectrometry." Thesis, Durham University, 2001. http://etheses.dur.ac.uk/3962/.
Full textAbstract:
Poly(methyl methacrylate) (PMMA) homopolymers synthesised using 'classical' anionic methods and subsequently studied by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF-MS) are discussed. Specifically, the attempts at different end-group functionalisation reactions, their varying degrees of success, and the characterisation of these functionalized polymers via MALDI are reported. Extra peaks were observed in the spectra of samples containing a tertiary amine end-group. A mechanism for the in situ elimination of H(_2)(g) involving these end-groups, which would fit the observations, is proposed. Two alternative, 'non-classical' routes to the desired materials were investigated, as difficulties in successfully performing capping reactions to give end functionalised PMMA were noted. The first method was a variation of standard anionic polymerisation that involved the use of lithium silanolates, which could be performed at a higher temperature than normal. The second was a controlled free-radical technique known as Reversible Addition-Fragmentation Chain Transfer (RAFT). A lack of control of the polymerisation to the desired degree was observed with the former method. A well-defined RAFT sample was observed to undergo in situ eliminadon also, for which a mechanism involving the dithioester end-group is proposed, and which is supported by MALDI-collision induced dissociation (CID) evidence. The synthesis of block copolymers of various compositions of MMA with r-butyl methacrylate (t-BMA) and hexyl methacrylate (HMA), along with their homopolymers, and their subsequent characterisation is reported. PHMA was analysed easily, in contrast to Pt-BMA. Only copolymers with a high PMMA content were analysed successfully and this has been rationalised in terms of the factors that affect cationisation. The characterisation of equimolar blends of various end-functionalised PMMA samples is reported also. Samples that favour the binding of a metal ion over protonation appear to have a higher ion yield. Once more, these observations are rationalised in terms of the factors that affect cationisation.
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Sandonato, Beatriz Brabetz. "Peptídeos cíclicos hepatotóxicos : MALDI-TOF como uma ferramenta analítica /." Rio Claro, 2014. http://hdl.handle.net/11449/123921.
Full textAbstract:
Orientador: Humberto Márcio Santos MilagreBanca: Paulo José Samenho Moran
Banca: José Augusto Rosário Rodrigues
Resumo: As cianobactérias são bactérias fotossintetizantes que podem ser encontradas nos mais variados ambientes. Algumas espécies de cianobactérias produzem toxinas denominadas cianotoxinas. As cianotoxinas denominadas microcistinas (MCs) são heptapeptídeos cíclicos hepatotóxicos produzidas durante florações de cianobactérias. Sua detecção e quantificação em mananciais são de grande importância devido aos danos a saúde humana que essas microcistinas podem causar. Atualmente, a técnica MALDI-TOF-MS tem se mostrado uma eficiente ferramenta para a análise de microcistinas, e recentes estudos tem reportado seu uso na quantificação destas. Diante do exposto, o objetivo desta dissertação foi a avaliação de diferentes matrizes e diferentes métodos de preparo de amostra para MALDI-MS visando a detecção e quantificação das microcistinas. Com a finalidade de alcançar o melhor método de quantificação das microcistinas, nove matrizes para MALDI, onze métodos de preparo de amostra mais um método que foi adaptado por nosso grupo de pesquisa, o método Vacuum drying adaptado, e dois padrões comerciais de microcistinas, MC-LR e MC-RR, foram utilizados. A avaliação dos métodos de preparo de amostra foi realizada utilizando como matriz o ácido α-ciano-4-hidroxicinâmico (HCCA) e o peptídeo angiotensina I como padrão interno. Os métodos foram avaliados com relação aos seus coeficientes de variação (CV), suas cristalizações e espectros obtidos, utilizando a MC-RR como amostra. O método Vacuum drying adaptado apresentou os melhores resultados e sua curva analítica foi construída utilizando ambas as variantes de microcistinas. O limite de detecção (MLD) e o limite de quantificação (LDQ) também foram calculados. Cada curva da variante de microcistina apresentou excelente linearidade e os valores de r variaram entre 0,98-99, demonstrando que o método é adequado para a quantificação destas. Após as análises dos...
Abstract: Cyanobacteria are photosynthetic bacteria that can be found in diverse environments. Some species of cyanobacteria produce toxins denominated cyanotoxins. The cyanotoxins called microcystins (MCs) are hepatotoxic cyclic heptapeptides produced during cyanobacterial blooms. Its detection and quantification in springs are of great importance due to damage to human health that these microcystins can cause. Currently, MALDI-TOF-MS technique has been shown to be an efficient tool for the analysis of microcystins, and recent studies have reported its use in the quantification of these. On the exposed, the aim of this thesis was to evaluate the different matrices and different methods of sample preparation for MALDI-MS aimed at the detection and quantification of microcystins. With the aim of achieving the best method of quantification of microcystins nine matrices for MALDI eleven methods of sample preparation over a method that was adapted by our research group, the adapted vacuum drying method, and two commercial standards of microcystins, MC-LR, and MC-RR, are used. The evaluation methods of sample preparation was performed using as matrix the α- cyano-4-hydroxycinnamic acid (CHCA) and the angiotensin I as internal standard. The methods were evaluated with respect to their coefficients of variation (CV), their crystallization and spectra obtained using the MC-RR as a sample. The adapted vacuum drying method showed the best results and their analytical curve was constructed using both variants of microcystins. The limit of detection (MDL) and the limit of quantification (LOQ) were also calculated. Each curve variant of microcystin showed excellent linearity and r values ranged from 0.98 to 99, showing that the method is suitable for quantifying these. After analysis of the methods of sample preparation, the nine matrices were evaluated with respect to CV, crystallization and spectra, using the MC-RR as a sample. The best results were obtained...
Mestre
29
Dempwolf, Wibke. "MALDI-TOF in der kontrollierten radikalischen Polymerisation und Präpolymeranalyse." Clausthal-Zellerfeld Papierflieger, 2007. http://d-nb.info/990376834/04.
Full text
30
Schürch, Stefan. "Analytik biologischer und synthetischer Polymere mittels MALDI-TOF Massenspektrometrie /." [S.l.] : [s.n.], 1995. http://www.ub.unibe.ch/content/bibliotheken_sammlungen/sondersammlungen/dissen_bestellformular/index_ger.html.
Full text
31
Siricord, Cornelia Charito. "Detection of Phytophthora species by MALDI-TOF mass spectrometry." Thesis, Siricord, Cornelia Charito (2005) Detection of Phytophthora species by MALDI-TOF mass spectrometry. PhD thesis, Murdoch University, 2005. https://researchrepository.murdoch.edu.au/id/eprint/314/.
Full textAbstract:
Phytophthora diseases have caused worldwide economic, social and environmental impacts for decades. Once their presence is confirmed, they are difficult to eradicate. To reduce and manage the damage inflicted by the pathogen, fast and reliable disease management protocols are required. Tests that enable the rapid and reliable identification of the pathogen assist greatly in disease management.
Phytophthora species are traditionally not only detected by baiting but also by plating of symptomatic tissue on selective media. Species can be identified by the characteristics of the mycelium growing out of the bait. However, the method is low throughput, labour intensive, and prone to false negatives. An alternative approach would be to detect the pathogen by the presence of its DNA. This involves amplification of the pathogen DNA using Polymerase Chain Reaction (PCR) and detection of the amplification product. Detection is usually by agarose gel electrophoresis. However, this is also a labour intensive process involving pouring, loading, running, and staining of the gels. The aim of this thesis is to explore the use of Matrix Assisted Laser Desorption/ Ionisation Time-of-Flight (MALDI-TOF) mass spectrometry for detection of PCR products. This procedure enables the analysis of large numbers of samples within a very short time-frame as the average time for analysis of each sample is in the order of milliseconds.
The assay involves annealing an extension (genotyping) primer to the PCR product and its extension by a single nucleotide. The nature of the nucleotide added differentiates species as does the site to which the primer anneals. Multiple extension (genotyping) primers can be used together in a single reaction for detection of multiple species. In this project four genotyping primers (GPs) were designed from the ITS regions of Phytophthora palmivora, Phytophthora cinnamomi, Phytophthora citricola, and Phytophthora cambivora.
The extension primers were tested for their specificity on the DNA of the target species. The four primers designed were specific for their intended targets except for GPpalm3 which in addition to being extended by ddT when tested with DNA from P. palmivora, was also extended by ddC when tested with DNA from other species of Phytophthora or Pythium.
These primers were also tested for their ability to detect multiple Phytophthora species in a single reaction (multiplexing). Mixtures of primers were added to mixed DNA templates and the primer extension reaction carried out. The primers were designed so that their masses were sufficiently different for them to be identified from a mixture. Six replicates were analysed for each reaction. In general only about 1-3 of the six replicates gave a positive reaction. This indicates that there may be some interference between primers, or that the presence of all four nucleotides interfered with the primer extension reaction. Increasing either the amount of enzyme, the amount of nucleotides or both did not improve the results.
The sensitivity of detection was tested by the addition of different amounts of mycelium to soil. The detection sensitivity depended on the primer pair used for PCR amplification. The ITS1/2 primer pair was more sensitive than the ITS1/4 pair. The limit of detection was 1 mcg mycelium g soil-1. However using nested PCR, levels of sensitivity comparable to those obtained using the ITS1/2 primer pair could be achieved. Primers to other regions of the genome such as the beta cinnamomin elicitin gene gave very low levels of sensitivity compared to the ITS primers.
In comparison with DNA detection we found that the limit of detection using baiting was 4 mcg mycelium g soil-1. Results below this limit were unreliable. The method suffered from the additional disadvantage that it took a long time in comparison to DNA detection.
DNA detection methods do not distinguish between living and dead organisms in the soil. However it can be hypothesised that DNA is unlikely to persist for any significant length of time in soil. To test this, we added plasmid DNA to soil and tested the persistence of this DNA using a variety of methods such as precipitation of labelled DNA, southern blotting and PCR amplification. It was found that in general, in soils from different ecosystems, the bulk of the DNA was undetectable after 24 hours. The rate of DNA breakdown differed with the soil type. In some soils, the added DNA was not detected even after 2 hours, whereas in others it could be observed after 10 hours. The detection depended on the method. Southern blotting showed that although DNA could be observed at 10 hours, by 24 hours it was completely degraded. In contrast a PCR product could be obtained from the soil extracts up to 24 hours. In a separate experiment, plasmid DNA was detectable over a 24 hour incubation period in 5 soil samples from 5 different sites. The results suggest that DNA is degraded rapidly in soil and is unlikely to persist longer than 24 hours.
The results in this thesis demonstrate that MALDI-TOF MS is a suitable alternative to agarose gel electrophoresis for analysis of PCR products. The technique is rapid, differentiates species from mixtures, is high-throughput and amenable to automation. Implementation will require further research to automate the primer extension assay to reduce the sensitivity to impurities in the DNA and to design parameters for sampling asymptomatic material.
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Siricord, Cornelia Charito. "Detection of Phytophthora species by MALDI-TOF mass spectrometry." Siricord, Cornelia Charito (2005) Detection of Phytophthora species by MALDI-TOF mass spectrometry. PhD thesis, Murdoch University, 2005. http://researchrepository.murdoch.edu.au/314/.
Full textAbstract:
Phytophthora diseases have caused worldwide economic, social and environmental impacts for decades. Once their presence is confirmed, they are difficult to eradicate. To reduce and manage the damage inflicted by the pathogen, fast and reliable disease management protocols are required. Tests that enable the rapid and reliable identification of the pathogen assist greatly in disease management.
Phytophthora species are traditionally not only detected by baiting but also by plating of symptomatic tissue on selective media. Species can be identified by the characteristics of the mycelium growing out of the bait. However, the method is low throughput, labour intensive, and prone to false negatives. An alternative approach would be to detect the pathogen by the presence of its DNA. This involves amplification of the pathogen DNA using Polymerase Chain Reaction (PCR) and detection of the amplification product. Detection is usually by agarose gel electrophoresis. However, this is also a labour intensive process involving pouring, loading, running, and staining of the gels. The aim of this thesis is to explore the use of Matrix Assisted Laser Desorption/ Ionisation Time-of-Flight (MALDI-TOF) mass spectrometry for detection of PCR products. This procedure enables the analysis of large numbers of samples within a very short time-frame as the average time for analysis of each sample is in the order of milliseconds.
The assay involves annealing an extension (genotyping) primer to the PCR product and its extension by a single nucleotide. The nature of the nucleotide added differentiates species as does the site to which the primer anneals. Multiple extension (genotyping) primers can be used together in a single reaction for detection of multiple species. In this project four genotyping primers (GPs) were designed from the ITS regions of Phytophthora palmivora, Phytophthora cinnamomi, Phytophthora citricola, and Phytophthora cambivora.
The extension primers were tested for their specificity on the DNA of the target species. The four primers designed were specific for their intended targets except for GPpalm3 which in addition to being extended by ddT when tested with DNA from P. palmivora, was also extended by ddC when tested with DNA from other species of Phytophthora or Pythium.
These primers were also tested for their ability to detect multiple Phytophthora species in a single reaction (multiplexing). Mixtures of primers were added to mixed DNA templates and the primer extension reaction carried out. The primers were designed so that their masses were sufficiently different for them to be identified from a mixture. Six replicates were analysed for each reaction. In general only about 1-3 of the six replicates gave a positive reaction. This indicates that there may be some interference between primers, or that the presence of all four nucleotides interfered with the primer extension reaction. Increasing either the amount of enzyme, the amount of nucleotides or both did not improve the results.
The sensitivity of detection was tested by the addition of different amounts of mycelium to soil. The detection sensitivity depended on the primer pair used for PCR amplification. The ITS1/2 primer pair was more sensitive than the ITS1/4 pair. The limit of detection was 1 mcg mycelium g soil-1. However using nested PCR, levels of sensitivity comparable to those obtained using the ITS1/2 primer pair could be achieved. Primers to other regions of the genome such as the beta cinnamomin elicitin gene gave very low levels of sensitivity compared to the ITS primers.
In comparison with DNA detection we found that the limit of detection using baiting was 4 mcg mycelium g soil-1. Results below this limit were unreliable. The method suffered from the additional disadvantage that it took a long time in comparison to DNA detection.
DNA detection methods do not distinguish between living and dead organisms in the soil. However it can be hypothesised that DNA is unlikely to persist for any significant length of time in soil. To test this, we added plasmid DNA to soil and tested the persistence of this DNA using a variety of methods such as precipitation of labelled DNA, southern blotting and PCR amplification. It was found that in general, in soils from different ecosystems, the bulk of the DNA was undetectable after 24 hours. The rate of DNA breakdown differed with the soil type. In some soils, the added DNA was not detected even after 2 hours, whereas in others it could be observed after 10 hours. The detection depended on the method. Southern blotting showed that although DNA could be observed at 10 hours, by 24 hours it was completely degraded. In contrast a PCR product could be obtained from the soil extracts up to 24 hours. In a separate experiment, plasmid DNA was detectable over a 24 hour incubation period in 5 soil samples from 5 different sites. The results suggest that DNA is degraded rapidly in soil and is unlikely to persist longer than 24 hours.
The results in this thesis demonstrate that MALDI-TOF MS is a suitable alternative to agarose gel electrophoresis for analysis of PCR products. The technique is rapid, differentiates species from mixtures, is high-throughput and amenable to automation. Implementation will require further research to automate the primer extension assay to reduce the sensitivity to impurities in the DNA and to design parameters for sampling asymptomatic material.
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Borsani, M. "BIOINFORMATICS APPROACHES TO MALDI-TOF MASS SPECTROMETRY DATA ANALYSIS." Doctoral thesis, Università degli Studi di Milano, 2013. http://hdl.handle.net/2434/221050.
Full textAbstract:
Despite the increasing performance of Mass spectrometry (MS) and others analytical tools, only few biomarkers have been validated and proved to be robust and clinically relevant; indeed a large numbers of proteomic biomarkers have been described, but they are not yet clinical implemented [1]. MALDI-TOF MS seems one of the more powerful tool for biomarkers discovery [2, 3], and shows interesting clinical properties, for instance the possibility to directly search in peripheral fuids for proteins related to an altered physiological state: samples (urine, plasma, serum, etc.) can be collected easily and cheaply by non-invasive, or very low-invasive, methods [4]. The combination of some biomarkers is actually considered more informative than a single biomarker [5, 6], and the improvement in the bioinformatics analysis of MS data could probably help this investigation, decreasing costs and time necessary for each discovery [7].
It is possible to approach the problems related to the analysis of (MALDI-TOF) MS data in two ways, either trying to increase the number of available samples or by reducing the complexity of the problem [8]: in the first case, we developed an approach to compare small datasets from different sources (i.e. hospitals), based on mutual information and mass spectra alignment, that showed significant performance increase compare to the competing ones tested.
In the latter case, we developed novel methods and approaches to compare MALDI-TOF MS profiles of normal and Renal Cell Carcinoma (RCC) patients, with the goal of isolating the more interesting subset of small proteins and peptides from the whole analysed peptidome. MS-based profiling is in fact able to detect differently expressed proteins or peptides during physiological and pathological processes. Every MALDI-TOF MS spectrum, that reports the relative abundance of sample analytes, could be considered as a snapshot of samples peptidome in a definite mass range. The relationship between mass/charge ratio, or m/z, and concentration of detected peptides can be represented by networks. Tumor case and control subjects show different peptidome profiles, due to differences in biomolecular and/or biochemical features of cancer cells: they will show some changes in the networks that describe them. We use graphs to create networks representation of data and to evaluate networks properties. We explore the networks properties comparing cases versus controls datasets, and subdividing cases in the different histological subtypes of RCC, clear cell RCC (ccRCC) and not-ccRCC, using different methods both for networks creation and analysis, and for results evaluation. We identify, for each datasets (controls, ccRCC and not-ccRCC) some interesting mass ranges within which we believe biomarkers signals should be searched.
In conclusion, we have developed a set of methods which we believe improve the current computational approaches for the analysis of mass spectrometry data. These results have been published or presented at workshops and conferences.
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Olafsson, Jonas. "Antimicrobial Susceptibility Testing Directly from Urine Samples : a Comparison between Standardised and Direct Disk Diffusion Testing together with Direct Species Identification using Matrix Assisted Laser Desorption/Ionisation Time of Flight." Thesis, Linnéuniversitetet, Institutionen för naturvetenskap, NV, 2012. http://urn.kb.se/resolve?urn=urn:nbn:se:lnu:diva-27645.
Full textAbstract:
Urinary tract infection (UTI) is a very common infection in humans and a majority is caused by Escherichia coli. UTI are commonly treated empirically. However, empiric treatment has become more problematic due to increased antibiotic resistance to commonly used antibiotic agents. It is therefore desirable with short turnover times for antimicrobial susceptibility testing and species identification to improve antibiotic treatment at an early stage. Matrix Assisted Laser Desorption/Ionisation Time of Flight (MALDI-TOF) can provide species identification faster than former routine methods. This study compared direct and standard susceptibility testing using disk diffusion on Enterobacteriaceae (EB) from urine samples. The possibility to standardise the inoculum for direct susceptibility testing via a pellet obtained by a series of centrifugations was also evaluated, as well as direct species identification with MALDI-TOF from the pellet. Results from direct susceptibility testing from urine samples with EB, performed either directly from the urine or with a standardised inoculum, correlated well to those obtained with standardised susceptibility testing using EUCAST disk diffusion methodology with few errors, of which most were associated with Proteus mirabilis. The concept of standardising the inoculum for direct susceptibility testing to 0.5 McFarland was labour intensive and did not improve the results further. However, direct species identification from the urine pellet using MALDI-TOF showed good correlation to routine identification. Of 238 samples, an EB was correctly identified in 148 samples using MALDI-TOF.
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Lourenço, Tarcísio Torre. "Estudo proteômico do desenvolvimento folicular de vacas zebuinas não gestantes." Botucatu, 2016. http://hdl.handle.net/11449/146671.
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Orientador: Fabiana Ferreira de SouzaResumo: O ciclo estral da vaca é composto por 2-3 ondas de crescimento folicular, no qual vários folículos são recrutados e iniciam um novo crescimento. Durante o período denominado desvio folicular, um folículo se torna dominante e os outros entram em atresia. Este processo envolve um mecanismo ainda não completamente compreendido, incluindo proteínas específico, como já estabelecido pela expressão gênica. O objetivo do presente estudo foi caracterizar as proteínas do fluído folicular a fim de identificar macromoléculas relacionadas ao desenvolvimento dos folículos de vacas zebuínas nã-gestantes. Foram colhidos os ovários de 25 vacas mestiças não-gestantes em um abatedouro. A presença do corpo lúteo foi anotada para cada ovário. O líquido folicular foi colhido utilizando-se a imersão do ovário em meio líquido e ultrassonografia. De acordo com a mensuração do diâmetro folicular, foram formados 3grupos, folículos pequenos (≤6,5mm, n=25), médios (>6,5mm a ≤9mm, n=9) e grandes (>9,0mm, n=11). Após 2 centrifugações (600xg/10 minutos e 15.000xg/30 minutos, 4ºC) o sobrenadante foi separado e utilizado para determinação da concentração de proteína total (método de Bradford). A eletroforese foi conduzida sob condições desnaturantes e redutoras, em gel de separação de poliacrilamidaà 12%. A concentração de progesterona e estradiol do líquido folicular foi determinada a fim de identificar os folículos saudáveis. As proteínas diferenciais identificadas pela eletroforese foram... (Resumo completo, clicar acesso eletrônico abaixo)
Mestre
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Glückmann, Matthias. "Untersuchungen zur Ablation und Ionenbildung bei matrixunterstützter Laserdesorption-Ionisation (MALDI)." [S.l. : s.n.], 2001. http://deposit.ddb.de/cgi-bin/dokserv?idn=963583107.
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Priyasantha, Kandalama KD. "DEVELOPMENT OF A NOVEL MATRIX ASSISTED LASER DESORPTION / IONIZATION (MALDI) BASED PEPTIDE QUANTITATION APPROACH." OpenSIUC, 2015. https://opensiuc.lib.siu.edu/dissertations/989.
Full textAbstract:
Matrix Assisted Laser Desorption / Ionization (MALDI) Mass Spectrometry (MS) has emerged as an important tool in the field of proteomics mainly because it is simple, quick and efficient. The identification and quantitation of biomarkers, protein targets for drugs, and metabolites are some of the important fields in proteomics research. Although MALDI MS is an important tool in proteomics research there are drawbacks of the technique that need further development in order for the approach to be used in clinical laboratories. One major limitation of MALDI MS is the generally poor reproducibility of ion signal intensities, which negatively impacts the quantitation of peptides and protein by MALDI MS. A considerable amount of research has been performed in an effort to improve the ion signal reproducibility in MALDI MS. However, many of the approaches developed have introduced specific drawbacks with respect to the traditional dried-droplet sample preparation technique, negating many of the advantages of the MALDI MS approach. This project has focused on the development of a novel approach to quantify peptides by MALDI MS while preserving traditional known advantages of the technique. The studies performed show that an approach in which the ion signal base widths are manipulated to match that of a reference ion signal, through adjustments in desorption laser intensity, leads to much higher reproducibility in the integrated ion signal intensities. A standard curve acquired using the constant ion signal base width approach showed lower average RSDs (< 10.00% vs.> 39.00%) and improved R2 values (> 0.9600 vs. < 0.809) as compared to the conventional constant desorption laser intensity approach. Subsequent work also revealed that the peptide hydrophobic / hydrophilic properties influenced the applicability of the quantitation approach to mixtures of peptides. Specifically, the data revealed that peptides with differing hydrophobic / hydrophilic properties appear to co-crystallize with the MALDI matrix differently leading to an inability to use a hydrophobic peptide signal to quantitate a hydrophilic peptide, and vice versa. This latter conclusion was further supported in similar studies performed on the mixture of peptides resulting from tryptic digestion of the protein bovine serum albumin.
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Crank, Jeffrey Aaron. "Ionic liquids MALDI-MS matrices and gas chromatography stationary phases /." [Ames, Iowa : Iowa State University], 2009. http://gateway.proquest.com/openurl?url_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&res_dat=xri:pqdiss&rft_dat=xri:pqdiss:3379180.
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Przybilla, Laurence. "Methodische Entwicklung der MALDI-TOF-Massenspektrometrie für Grenzbereiche der Polymeranalytik." [S.l. : s.n.], 2000. http://ArchiMeD.uni-mainz.de/pub/2000/0091/diss.pdf.
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Trimpin, Sarah. "Methodische Entwicklung der MALDI-TOF-Massenspektrometrie für Grenzbereiche der Makromolekülanalytik." [S.l. : s.n.], 2002. http://deposit.ddb.de/cgi-bin/dokserv?idn=971568650.
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Hellwig, Nils [Verfasser]. "Das Laserablationsverhalten von ionischen Flüssigkeiten verschiedener MALDI-Matrices / Nils Hellwig." Kiel : Universitätsbibliothek Kiel, 2012. http://d-nb.info/1025465121/34.
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Allwood, Daniel Anthony. "Characterisation and ionisation modelling of matrices in MALDI mass spectrometry." Thesis, University of Hull, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.301477.
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Schumacher, Joshua. "Carbon Nanotube Enhanced MALDI MS: Increasing Sensitivity Through Sample Concentration." Scholar Commons, 2010. http://scholarcommons.usf.edu/etd/3595.
Full textAbstract:
Matrix-assisted laser desorption/ionization (MALDI) is a technique used in mass spectrometry for the ionization of biomolecules. A matrix solution is mixed with the analyte molecules to be investigated, and then spotted onto a specialized MALDI plate. The solvents evaporate leaving only the re-crystallized matrix with analyte dispersed throughout the crystals. Sample ionization is accomplished with a laser in the MALDI instrument. The spot diameter of the target is usually several orders of magnitude larger than the diameter of the laser, making it necessary to perform multiple laser investigations to accurately evaluate the analyte in the target spot.
Experiments were performed to utilize patterned areas of carbon nanotubes to provide sites for preferential crystallization of the liquid matrix/analyte solution, which led to lateral concentration for non-aqueous based matrices and produced a final dried matrix/analyte spot that was approximately the diameter of the laser spot at the point of investigation. This work shows the results of using aligned carbon nanotubes as the substrate for the matrix/analyte deposition and demonstrates an increase in signal to noise ratio and an improved detection capability of low analyte concentrations compared to the standard MALDI preparation technique.
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Worster, Belinda Mary. "MALDI-TOF-MS and neuropeptide signalling in the nervous system." Thesis, University of Sussex, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.360682.
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Steven, Rory Thomas. "Investigations in MALDI-MSI using a high repetition rate laser." Thesis, University of Birmingham, 2014. http://etheses.bham.ac.uk//id/eprint/5392/.
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Within these studies the properties of a high repetition rate (≤ 25,000 Hz) Nd:YVO4 laser were investigated with a view to improving the throughput and quality of information obtained from MALDI MS data collected in continuous raster sampling mode. Initially, the nature of the laser beam profile was investigated and a novel fluorometric method developed for imaging these profiles. Analysis of thin film samples of α-cyano-4-hydroxycinnamic acid (CHCA) and lipid standard phophatidylcholine (PC) 34:1 were carried out. Under most conditions a lower repetition rate and slow stage raster speed were found to be optimal. However, in tissue based investigations repetition rates of 5-10 kHz and faster raster speeds were found to increase the detected ion intensity. Subsequent to this, the use of para-nitroaniline (PNA) as an effective matrix for high-repetition rate laser MALDI MSI applications was investigated and compared to CHCA. PNA was found to provide high quality MSI data, comparable to or better than that obtained with CHCA. The utility of CHCA and PNA were then investigated for the repeat analysis of single tissue sections with a view to increasing the amount of information obtained. Up to five analyses of the same tissue section were demonstrated. Repeat analysis was then applied to the acquisition of both lipid and protein data from a single tissue section using multiple matrices and tissue washing.
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Guan, Bing. "Characterization of Oligosaccharides and Nanoparticles by MALDI-TOF Mass Spectrometry." ScholarWorks@UNO, 2007. http://scholarworks.uno.edu/td/585.
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The possibilities of differentiating linkage positions and anomeric configurations of small oligosaccharides by negative ion mode MALDI using anion attachment followed by PSD are investigated. By careful initial adjustment of the focusing mirror ratios allowing acquisition of the peaks of interest within the same PSD segment, it is possible to obtain highly reproducible relative ion abundances. Discrimination of different linkage types is achieved by analysis of structurally-informative diagnostic peaks offered by PSD spectra of chloride adducts of oligosaccharides, whereas the relative peak intensities of selected diagnostic fragment pairs make differentiation of anomeric configuration possible. F- and Ac- cannot form anionic adducts with the oligosaccharides in significant yields. However, Br-, I- and NO3- anionic adducts consistently appear in higher abundances relative to [M - H]-, just like Cl-. Mildly acidic saccharides form both deprotonated molecules and anionic adducts, making it possible to simultaneously detect neutral and acidic oligosaccharides via anion attachment. PSD of [oligosaccharide + Cl]- yields structurally-informative fragment ions that retain the charge on the sugar molecule rather than solely forming Cl-, whereas PSD of Br-, I- and NO3- adducts of oligosaccharides yield the respective anions as the main product ions without offering structural information concerning the sugar. PSD of the chloride adduct of saccharides containing 1-2 linkages also yields chlorine-containing fragment ions. MALDI-TOF-MS and LDI-TOF-MS are shown to be useful for characterization of ultra-small titania nanoparticles. Peak maxima in MALDI-TOF mass spectra are found to correlate with nanoparticle size. The size distributions of TiO2 nanoparticles, obtained from MALDI- and LDI-TOF-MS are in good agreement with parallel TEM observations. PSD analysis of inorganic x nanomaterials is performed and valuable information about the structure of analytes has been obtained. A group of inorganic nitrate and perchlorate salts of forensic and health interest are investigated by LDI- and MALDI-TOF MS. In each case, a series of characteristic cluster ions are predominant in the negative-ion mode. The number and identity of metal atoms and anions in the recorded cluster ions can be positively identified by their m/z values, distinctive isotopic patterns and characteristic PSD fragmentation patterns.
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Börjesson, Erik. "Undersökning av jonvätskematrisers detektion på olika typer av MALDI plattor." Thesis, KTH, Skolan för kemivetenskap (CHE), 2016. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-195847.
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Examensarbetet gjordes för att undersöka om det istället för 2,5-dihydroxidbensoesyra fanns en jonvätskematris som hade en lika bra eller bättre detektionsgrad. Examensarbetet undersökte också om det var någon detektions skillnad på stål- och koncentrationsplatta när analysmetoden matris-assisterande laser desorption-jonisation-masspektrometri (MALDI-MS) analysmetod används. Jonvätskematris är en matris som binder till en svag bas och är till för att skydda peptiderna från addukter. Baserna som undersöktes var etylamin, dietylamin, diisopropylamin och N,N-etyl-diisopropylamin. Examensarbetet jämförde matrisen med jonvätskematriserna som alla var upplösta i etanol, vatten eller en vätskeblandning (TA 30) (0,1 % trifluorättiksyra i vatten och acetonitril i 7:3 volym). Det utfördes tester genom att matrisen och jonvätskematriserna blandades med en peptidblandning innehållande åtta stycken peptider. Blandningen deponerades på en stål- och en koncentrationsplatta. Efter att blandningen hade torkat på plattorna och bilde till kristaller eller en vätskefilm placerades plattorna i MALDI-apparaturen. Testerna utvisade att N,N-etyl diisopropylamin blandningen i etanol och vätskeblandningen var bättre än 2,5-dihydroxybenzene på att detektera antalet peptider på koncentrationsplattan. Det visades att etylamin och dietylamin hade samma detektiongrad som 2,5-dihydroxidbensoesyra i etanol och vatten vid detektion av antal peptider på koncentrationsplattan. En detektionsskillnad fanns mellan stål- och koncentrationsplatta. Stålplattan var bättre på att detektera tyr-bradykinin. Vid vidare studier på området skulle testerna utföras på stålplatta för att detektera tyr-bradykinin och resterande peptider på koncentrationsplatta. Vid vidare studier skulle TA 30 och etanol som lösningsmedel användas och matriserna ska upplösas i mol-koncentration istället för vikt-koncentration, för att få en mer jämförbar detektion.This degree project was made to examine if there was a better ionic liquid matrix instead of the matrix 2,5-dihydroxybenzene. The aim of the project was also to investigate if there were any detection differences between anchorchip and groundsteel plate when testing matrix assisted laser desorption ionization masspectrometry (MALDI-MS). An ionic liquid matrix is a matrix that binds to a base used to protect the peptides from adducts. In the degree project 2,5-dihydroxybenzene was compared with the ionic liquid matrices which is 2,5-dihydroxybenzene combined with a base. The bases were ethylamine, diethylamine, diisopropylamine and N,N- ethyldiisopropylamine. All matrices were dissolved in ethanol, water or a liquid mix (0,1 % Trifluoroacetic acid in water and acetonitril in 7:3 volume). The tests were performed by mixing the matrix and the ionic liquid matrices with a peptide mix which contained 8 types of peptides. The matrix/ionic liquid matrices and peptide mix was deposited on a ground steel and anchorchip MALDI plate. After the mixture had evaporated and transformed to crystals/film, the plates were put in MADLI equipment. The results showed that N,N- ethyldiisopropylamine in ethanol and liquid mix had better detection than 2,5-dihydroxybenzene. Ethylamine and diethylamine had the same detection as 2,5-dihydroxybenzene in ethanol on anchorchip plate. There were detection differences between the anchorchip and ground steel plate. In favor of ground steel plate when detecting tyr-bradykinin, but for detecting the rest of the peptides anchorchip is favorable. If further studies would be performed in this area it would be done in the liquid mix and ethanol as solvent. For more comparable detection results the matrices concentration should be in mol/L instead of g/L.
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Bronzel, Júnior João Luiz [UNESP]. "Matrizes iônicas: detecção e quantificação de cianotoxinas por maldi-ms." Universidade Estadual Paulista (UNESP), 2015. http://hdl.handle.net/11449/143001.
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000854199.pdf: 5245942 bytes, checksum: 78603c065ca6ba2cb04fe0531fde9643 (MD5)A técnica de ionização MALDI (Matrix-Assisted Laser Desorption/Ionization) foi uma das responsáveis por revolucionar a Espectrometria de Massas, tornando possível sua aplicação a moléculas termolábeis e de alta massa molecular, como as proteínas. Apesar de seu grande desenvolvimento, esta técnica ainda apresenta algumas limitações como a seleção da matriz utilizada e o método de preparo de amostras. Estes fatores são determinantes na reprodutibilidade da análise e na faixa de aplicação de m/z. Neste trabalho, inicialmente analisou-se diferentes linhagens de cianobactérias por MALDI-TOF-MS, com a finalidade de se buscar por novos analitos para os quais fosse possível o monitoramento utilizando-se as matrizes iônicas. Posteriormente avaliou-se as características destas matrizes, a metodologia de preparo de amostras, a composição do analito e a intensidade do laser da fonte de MALDI, com o objetivo de selecionar as melhores metodologias de análise. Após esta etapa, as matrizes iônicas e metodologias selecionadas foram aplicadas na detecção e quantificação de cianotoxinas e na análise de fármacos por MALDI-MS. Neste estudo obteve-se como resultados: a diferenciação de linhagens de cianobactérias através dos fingerprints obtidos por MALDI-TOF-MS que também permitiram a detecção de importantes metabólitos; a detecção da homoanatoxina-a, uma cianotoxina de baixa massa molecular, diretamente de células de cianobactérias; o desenvolvimento de um método quantitativo de análise de microcistinas com ótima precisão; e o desenvolvimento de uma metodologia inédita para a detecção dos componentes ativos de medicamentos, ampliando, portanto, o leque de aplicações da técnica de MALDI.
The Matrix-Assisted Laser Desorption/Ionization technique was one of the ionization sources responsible for revolutionizing Mass Spectrometry, making possible its application to labile and high molecular weight molecules, such as proteins. Despite its great development, this technique still has some limitations like the selection of the matrix and the sample preparation method. These factors determine the reproducibility of analysis and the m/z application range. Initially, in this work, we analyzed different cyanobacteria strains by MALDI-TOF-MS, in order to check for new analytes to be monitored using ionic matrices. Subsequently, the characteristics of these matrices, the sample preparation method, the composition of the analyte and laser intensity of the MALDI source were evaluated aiming to select the best methodologies of analysis. After this step, the selected ionic matrices and methodologies have been applied in the detection and quantification of cianotoxins and analysis of pharmaceutical drugs by MALDI-MS. In this study we obtained as results: the differentiation of strains of cyanobacteria through the fingerprints obtained by MALDI-TOF-MS which allowed the detection of important metabolites; the detection of homoanatoxin-a, a low molecular weight cyanotoxin, directly from cyanobacteria cells; the development of a quantitative method for the analysis of microcystins with great precision; and the development of a new methodology for the detection of active compounds of medicines, increasing, therefore, the application range of MALDI technique.
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Bronzel, Júnior João Luiz. "Matrizes iônicas : detecção e quantificação de cianotoxinas por maldi-ms /." Araraquara, 2015. http://hdl.handle.net/11449/143001.
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Orientador: Humberto Márcio Santos MilagreBanca: Eduardo Maffud Cilli
Banca: José Augusto Rosário Rodrigues
Resumo: A técnica de ionização MALDI (Matrix-Assisted Laser Desorption/Ionization) foi uma das responsáveis por revolucionar a Espectrometria de Massas, tornando possível sua aplicação a moléculas termolábeis e de alta massa molecular, como as proteínas. Apesar de seu grande desenvolvimento, esta técnica ainda apresenta algumas limitações como a seleção da matriz utilizada e o método de preparo de amostras. Estes fatores são determinantes na reprodutibilidade da análise e na faixa de aplicação de m/z. Neste trabalho, inicialmente analisou-se diferentes linhagens de cianobactérias por MALDI-TOF-MS, com a finalidade de se buscar por novos analitos para os quais fosse possível o monitoramento utilizando-se as matrizes iônicas. Posteriormente avaliou-se as características destas matrizes, a metodologia de preparo de amostras, a composição do analito e a intensidade do laser da fonte de MALDI, com o objetivo de selecionar as melhores metodologias de análise. Após esta etapa, as matrizes iônicas e metodologias selecionadas foram aplicadas na detecção e quantificação de cianotoxinas e na análise de fármacos por MALDI-MS. Neste estudo obteve-se como resultados: a diferenciação de linhagens de cianobactérias através dos fingerprints obtidos por MALDI-TOF-MS que também permitiram a detecção de importantes metabólitos; a detecção da homoanatoxina-a, uma cianotoxina de baixa massa molecular, diretamente de células de cianobactérias; o desenvolvimento de um método quantitativo de análise de microcistinas com ótima precisão; e o desenvolvimento de uma metodologia inédita para a detecção dos componentes ativos de medicamentos, ampliando, portanto, o leque de aplicações da técnica de MALDI.
Abstract: The Matrix-Assisted Laser Desorption/Ionization technique was one of the ionization sources responsible for revolutionizing Mass Spectrometry, making possible its application to labile and high molecular weight molecules, such as proteins. Despite its great development, this technique still has some limitations like the selection of the matrix and the sample preparation method. These factors determine the reproducibility of analysis and the m/z application range. Initially, in this work, we analyzed different cyanobacteria strains by MALDI-TOF-MS, in order to check for new analytes to be monitored using ionic matrices. Subsequently, the characteristics of these matrices, the sample preparation method, the composition of the analyte and laser intensity of the MALDI source were evaluated aiming to select the best methodologies of analysis. After this step, the selected ionic matrices and methodologies have been applied in the detection and quantification of cianotoxins and analysis of pharmaceutical drugs by MALDI-MS. In this study we obtained as results: the differentiation of strains of cyanobacteria through the fingerprints obtained by MALDI-TOF-MS which allowed the detection of important metabolites; the detection of homoanatoxin-a, a low molecular weight cyanotoxin, directly from cyanobacteria cells; the development of a quantitative method for the analysis of microcystins with great precision; and the development of a new methodology for the detection of active compounds of medicines, increasing, therefore, the application range of MALDI technique.
Mestre
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Earnshaw, Caroline Jane. "Sample preparation methodologies for MALDI-MS imaging and related topics." Thesis, Sheffield Hallam University, 2009. http://shura.shu.ac.uk/19590/.
Full textAbstract:
The diverse applications of MALDI MSI are explored in this thesis with an emphasis on the sample preparation procedure and method development for small molecule analysis for a range of samples. The two main themes that have been focussed on are the pharmaceutical and metabolomic applications of this state of the art technique. MALDI MSI has been evaluated as a technique for the detection and imaging of antiasthmatic compounds in lung tissue. Four compounds were assessed initially with conventional MALDI MS experiments, followed by both direct and indirect tissue imaging experiments. Pharmaceutical tablet formulations have also been assessed using MALDI MSI to map the active component throughout the excipients contained within the tablet providing information that is critical to the manufacturing process such as the homogeneity of the active pharmaceutical ingredient (API) throughout the tablet. MALDI MSI has been applied to the relatively new addition to the 'omics sciences, metabolomics. A non-targeted metabolomics approach has been used to study both plant and animal tissue in an attempt to gain a greater understanding of the complex biological processes that occur within both types of tissue. Wheat grain was used as the model system to conduct the experiments and evaluate the application of both UV MALDI MS and IR LDI MS for plant metabolomics. These techniques provided complementary information to published literature, however the novel aspect of this study was the incorporation of imaging experiments for UV MALDI MS; this allowed the metabolites to be visualised in the wheat grain section. MALDI MSI was also used to explore the differences between mice with chronic relapsing experimental autoimmune encephalomyelitis; the animal model of multiple sclerosis alongside healthy controls. Spinal cord samples were analysed and the main difference was tentatively attributed to choline levels.