Browsing by study line "Kemi"

Bioorthogonal chemistry and click chemistry have gained tremendous attention during the past few years. They do not refer to a single reaction but to a class of reactions that take inspiration from nature. Click reactions are driven by a strong thermodynamic driving force and therefore they proceed via well-controlled and consistent reaction pathways. Click reactions afford specific products in high yields with negligible by-products. Bioorthogonal chemistry builds on the boundaries set by click chemistry. Bioorthogonal reactions can occur within a living system without interacting or interfering with the natural biological processes therefore both the reactants and the products must be inert and stable under physiological conditions. Bioorthogonal reactions have allowed the real-time study of several biomolecules such as glycans, lipids, nucleic acids, and proteins within living systems without cytotoxicity. In the literature section of the thesis, the most important physical properties of both the isonitrile and chlorooxime are introduced and the most important bioorthogonal reactions for both functionalities are highlighted. The bioorthogonal isonitrile-chlorooxime ligation is discussed in more detail and an example is given of how the ligation can be used to label the cell membrane of living cells with fluorescent moieties. To install a moiety on the cell membrane it must first be modified with small non-natural chemical functionalities also called “chemical reporters.” which can be installed with metabolic oligosaccharide engineering (MOE) using monosaccharide analogues. After installation of the “chemical reporters” onto the cell surface, molecules containing the corresponding bioorthogonal counterpart can be attached to the cell membrane. Lastly, the sulfur(VI) fluoride exchange (SuFEx) click chemistry is discussed. Traditionally, SuFEx click chemistry has been used in organic chemistry to build inorganic connecting bridges between two carbon centres. More recently, SuFEx chemistry has found utility in the radiosynthesis of fluorine-18 containing [18F]sulfonyl fluorides and [18F]fluorosulfates. Fluorine-18 is one of the most commonly used and important positron emitters utilized in radiopharmaceutical chemistry and positron emission tomography (PET). The experimental section of the thesis presents the synthesis routes of the bioorthogonal reaction partners, a peracetylated isonitrilepropanoylmannosamine (Ac4ManNC) and an aryl fluorosulfate chlorooxime. The research hypothesis of the study was that the isonitrile of the Ac4ManNC could be installed onto the cell surface of living Jurkat cells (human T lymphoblast) with MOE. Afterwards, the fluorine-18 labelled aryl [18F]fluorosulfate chlorooxime could have been attached to the cell surface from the isonitrile with the bioorthogonal isonitrile-chlorooxime ligation. This cell surface labelling method could have then been used in the future to research and develop cell therapy treatments by utilizing PET imaging.

Kultakomplekseille on tyypillistä typpi- ja rikkidonoreita sisältävät ligandit. Suurin osa tunnetuista kultakomplekseista on homoleptisiä, eli niissä kulta on liittynyt vain yhdentyyppisiin ligandeihin. Heteroleptisissä komplekseissa keskusatomiin on liittynyt useammanlaisia ligandeja. Heteroleptiset kompleksit ovat kiinnostavia, sillä kompleksien rakenteen muutokset vaikuttavat mm. katalyyttien tehokkuuteen. Kahden erilaisen ligandin avulla voidaan esimerkiksi vaikuttaa keskusatomin koordinaatiokykyyn merkittävästi. Kirjallisuuskatsauksessa tarkastellaan kullan komplekseja ja erilaisten ligandien käyttäytymistä kullan kanssa. Kirjallisuudesta löytyneiden heteroleptisten kultakompleksien ominaisuuksia, synteesireittejä ja käyttökohteita on esitelty. Kirjallisuudesta on etsitty esimerkkejä heteroleptisistä kultakomplekseista sekä kultatiolaattien reaktioista. Kokeellisessa osuudessa tutkittiin kullan liuotusreaktiota sekä pyrittiin valmistamaan ja analysoimaan heteroleptistä kultatiolaattia, jollaisia ei kirjallisuuskatsauksen perusteella ole valmistettu. Kullan liuotusreaktiossa, jossa hyödynnetään kahta eri tiolaattiligandia, on havaittu muodustuvan välituotteena heteroleptinen tiolaattikompleksi, jonka tarkempi karakterisointi oli työn tavoitteena. Vaikka valitulla synteesistrategialla kompleksia ei saatu eristettyä puhtaana, sen muodostuminen varmistui mm. massaspektroskopian avulla.

Ihmistoiminnan seurauksena luonnonvesiin päätyy haitallisia metalli-ioneja, joiden aiheuttamista ongelmista kärsivät nykyään eniten köyhät ja kehittyvät maat. Metalli-ionianalytiikalla on tärkeä rooli sekä luonnonvesien tarkkailussa että tutkimisessa kuin myös teollisten prosessien laadunvalvonnassa ja kehityksessä. Induktiivisesti kytketystä plasma-massaspektrometrista (ICP-MS) on tullut yksi yleisimmistä metallianalytiikassa käytettävistä analyysilaitteista. Useiden metalli-ionien pitoisuudet vesinäytteissä ovat kuitenkin laitteen määritysrajaa matalampia, jonka vuoksi analyytit täytyy esikonsentroida mittausta varten. Lisäksi suuri liuenneen kiintoaineen määrä saattaa vahingoittaa ICP-MS -laitetta. Edellä mainitut ongelmat ovat usein ratkaistavissa kiinteäfaasiuuton avulla, josta on kehittynyt erittäin monipuolinen ja helppo esikäsittelymenetelmä erilaisten vesinäytteiden analysointiin. Tutkielman kirjallisessa osiossa perehdytään metalli-ionianalytiikan merkitykseen sekä tutkituimpien metalli-ionien ominaisuuksiin ja terveysvaikutuksiin. Osiossa esitellään yleisimpiä ja uusimpia kiinteäfaasiuuttotekniikoita sekä vertaillaan off-line- ja on-line-sovellusten erityispiirteitä. Lisäksi perehdytään tekijöihin, jotka saattavat vaikeuttaa tai haitata kiinteäfaasiuuttoprosessia ja ICP-MS -mittausta. Kirjallisen osion loppupuolella käsitellään mielenkiintoisimpia, pääsääntöisesti vuosina 2010-2022, raportoituja kiinteäfaasiuuttosorbentteja, joita on käytetty tutkimuksissa, joissa mittaus on suoritettu ICP-MS:lla. Tutkielmassa käsitellään eräiden sorbenttien ja analyysiprosessien erityispiirteitä sekä tarkastellaan tärkeimpiä analyysiparametreja. Tutkielman kokeellisessa osiossa määritettiin hopea- ja koboltti-ionipitoisuuksia ICP-MS:lla Loviisan ydinvoimalaitoksen boorihappopitoisesta primääripiirin jäähdytteestä. Ydinreaktorissa aktivoituvat hopea- ja koboltti-ionit aiheuttavat merkittävän osan voimalaitostyöntekijöiden vuotuisesta säteilyannoksesta, jonka vuoksi kyseisten ionien tarkkailulla on suora vaikutus laitosturvallisuuteen. Määritystä varten boorihappo poistettiin aluksi selektiivisellä N-metyyliglukamiinipohjaisella ioninvaihtohartsilla. Tämän jälkeen hopea- ja koboltti-ionit esikonsentroitiin iminodiasetaattipohjaisella kiinteäfaasiuuttosorbentilla. Tulosten perusteella kehitettiin esikäsittelylaittoisto, joka koostui kahdesta päällekkäin asetetusta kolonnista. Metalli-ionipitoisuudet määritettiin ICP-MS:lle kehitetyn menetelmän avulla.

Nukleiinihapot ovat luonnollisia yksi- tai kaksisäikeisiä polymeerejä, jotka koostuvat deoksiribo- tai ribonukleosideistä linkitettynä toisiinsa fosfodiesterisidoksella. Kun tällaisia ketjuja valmistetaan kemiallisin menetelmin fosfaattiryhmä olisi aktivoitava tietyllä tavalla ja funktionaaliset ryhmät, jotka eivät osallistu reaktioon, olisi suojattava väliaikaisesti tai pysyvästi. Kiinnostus nukleiinihappokemiaan johtuu synteettisten oligomeerien ja niiden analogien kasvavasta tarpeesta välttämättöminä tutkimusvälineinä molekyylibiologiassa ja lääketieteessä. Tutkielman kirjallisessa osassa esitettiin erilaisia menetelmiä lyhyiden oligonukleotidien kemialliselle synteesille. Fosfodiesterisidosten muodostuminen tapahtuu yleensä joko fosfotriesterin tai fosfiitti-triesterin välituotteiden avulla. P(III)-välituotteiden suuremman reaktiivisuuden vuoksi fosfiitti-triesterimenetelmä ja erityisesti fosforamidiittimenetelmä ovat herättäneet huomiota. Oligonukleotidisynteesin lähtöaineiksi on ehdotettu useita erilaisia nukleosidi-fosforamidiitteja, kun on etsitty tasapainoa stabiilisuuden ja reaktiivisuuden välillä. Tämän vuoksi H-fosfonaattimenetelmää, jossa yhdistetään sekä fosfotriesteri- että fosfiitti-triesterimenetelmien edut ja lisäksi fosfodiesterimenetelmän edut (esim. fosforikeskuksen suojaavan ryhmän puute), voidaan käyttää vaihtoehtona fosforamidiittimenetelmälle erityisesti RNA:n sekä hapoille labiilien oligonukleotidianalogien synteesissä. Kaikilla menetelmillä on kuitenkin hyvät puolet sekä huonot puolet, joten ei olisi vielä olemassa yleisesti sovellettavaa ja tehokasta synteesimenetelmää, vaan ne sopivat eri tapauksiin. Esimerkiksi suuren mittakaavan synteesin tapauksessa tulee ottaa huomioon reaktioaika, reagenssi, puhdistusmenetelmä ja muut resurssit. Lisäksi lähestymistavat ovat yleensä joko erittäin työläitä etenkin lopputuotteen puhdistuksessa tai monivaiheisia sekvenssejä, joiden kokonaistuotto on alhainen. Kokeellisessa osassa valmistettiin kolmenlaisina Brønsted happokatalyytteinä pentakarboksisyklopentadieenit (PCCP). Tutkittiin mahdollisuutta käyttää PCCP-johdonnaisia regioselektiiviseen nukleosidin 5’-hydroksyyliryhmän suojaamiseen asetaaliryhmällä. Menetelmällä onnistuttiin valmistamaan tyydyttävä määrä 5’-O-asetaalisuojattua tymidiiniä. Menetelmä vaikuttaa lupaavalta pienellä jatkokehityksellä.

Piperine represents the major plant alkaloid encountered in various Piperaceae species and has received in recent years considerable attention because of its broad range of favorable biological and pharmacological activities, including antioxidant, immunostimulant, bioavailability-enhancing and anti-carcinogenic properties. The literature part of this thesis gives a selective overview of advanced methods for the quantitative analysis of piperine in plant-base materials, and various approaches employed for instrumental analysis, including spectroscopic, chromatographic, and electrochemical techniques. An effort was made to evaluate the potential of the reported methods based on the analytical figures of merit, such as total sample throughput capacity, analytical range, precision, accuracy, limit of detection and limit of quantification. The objective of the experimental part of the thesis focused on the development of a convenient, robust, simple, efficient and reliable method to quantify piperine in pepper fruits. The analytical method established in this thesis involves liberation of piperine by continuous liquid extraction of ground pepper fruits with methanol, and cleanup of the crude extracts with reversed phase solid phase extraction. Analyte quantitation was accomplished using gradient reversed phase High Performance Liquid Chromatography with mass spectrometric detection, using Electrospray Ionization-Ion Trap Mass Spectrometry. To enable reliable internal standardization, deuterium labelled piperine surrogate (piperine-D10) was synthesized from piperine in three steps in a reasonable overall yield (65 %) and standard-level purity (99.7 %). It may be worth mentioning that the commercial market value of the amount of piperine-D10 synthesized in-house exceeds 167,400 euros. One of the major challenges encountered during the development and optimization of the analytical method was the extreme photosensitivity of piperine and piperine-D10, both suffering in solution extensive photoisomerization upon exposure to ambient light within matter of minutes. This issue was addressed by carrying out all tasks associated with synthesis, sample preparation and analytical measurements under dark conditions. For the preparation of calibrators, a fully automated procedure was developed, being controlled by custom-written injector programs and executed in the light-protected sample compartment of a conventional autosampler module. In terms of merits, the developed analytical method offers good sample throughput capacity (run time 20 min, retention time 8.2 min), excellent selectivity and high sensitivity (Limit of Detection= 0.012 ppm, Limit of Quantification= 0.2 ppm). The method is applicable over a linear range of 0.4 to 20 ng of injected mass (r2= 0.999). The stability of standards and fully processed samples was found to be excellent, with less than 5% of variations in concentrations occurring after a 3-week (calibrators) or 4-month (samples) storage at 4 °C and 23 °C respectively, under dark conditions. Intra-day repeatability were better than 2.95 %. Preliminary validation data also suggest satisfactory inter-operator reproducibility. To test the applicability of the developed LC-MS method, it was employed to quantify piperine in a set of 15 pepper fruit samples, including black, white, red and green varieties of round and long peppers, purchased from local markets and retailers. The piperine contents obtained were in the range of 17.28 to 56.25 mg/g (piperine/minced sample) and generally in good agreement with the values reported in the scientific literature. It is justified to assume that the developed analytical method may directly be applicable to the quantitation of related pepper alkaloids in herbal commodities, and after some modifications in the sample preparation strategy, also for the monitoring of piperine in biological fluids, such as serum and urine.

The literature part of the thesis focused on reviewing published LC–MS/MS methods for cardiovascular drugs in biological matrices. Emphasis was given to the used sample preparation procedures and usability in different types of analyses, and their effects to the performances of the whole methods. Published methods for cardiovascular drugs in biological matrixes from the last decade were reviewed and presented. Insight into the challenges biological matrixes present was given, as well as a short overview of cardiovascular drugs. Plasma and urine were the most common sample matrixes, but some methods have also been published for serum and whole blood, and one even for fat tissue. Development and basic principles of liquid chromatography and mass spectrometry were presented, focusing on high and ultra-high performance liquid chromatography (HPLC and UHPLC) and triple quadrupole tandem mass spectrometry (QqQ–MS), as these seemed to be prevalent industry standard. Of sample preparation techniques, protein precipitation, direct dilution, liquid–liquid extraction (LLE), solid-phase extraction (SPE) and turbulent flow chromatography (TFC) were covered, with additions of some miniaturized versions. Some newer miniaturized online techniques have successfully been utilized for cardiovascular drug analysis, and their popularity will likely increase as the hardware in laboratories gets more and more modern. Advances in sample preparation techniques were relatively mild during the decade covered in this review, despite the significant proportion cardiovascular drugs hold in used medications. Newly published methods were either quantitative for a handful of analytes, or qualitative screenings for several analytes, some of which are cardiovascular drugs. Plasma and urine samples were used almost exclusively. In addition to the literary review, an experimental study was done at THL, where a LC–MS/MS method was successfully developed and validated during 2018–2019. The method has been used since 2019 as the primary qualitative method in cause–of–death investigations for 55 cardiovascular drugs analysed from post-mortem whole blood samples. In light of the done review, this method is exceptionally comprehensive regarding included analytes, and could potentially be at least partially automated with suitable instruments

The Venusian atmosphere has everything to be an exciting natural sulfur laboratory. In addition to relatively high concentrations of sulfur dioxide, suitable conditions in the atmosphere make both thermo- and photochemical reactions possible, allowing for complex chemical reactions and the formation of new sulfur containing compounds. These compounds could explain or contribute to the enigmatic 320-400 nm absorption feature in the atmosphere. One of the proposed absorbers is polysulfur compounds. While some experimentally obtained UV-VIS spectra have been published, studying the different polysulfur species individually is extremely difficult due to the reactive nature of sulfur. In this thesis UV-VIS spectra for polysulfur species S2 to S8 were simulated using the nuclear ensemble approach to determine if they fit the absorption profile. In total, 38 polysulfur species were considered. All were optimized at the wB97X-D/aug-cc-pV(T+d)Z level of theory, with the S2, S3, and S4 structures also being optimized at the CCSD(T)/aug-cc-pV(T+d)Z level of theory. For 13 structures UV-VIS spectra were simulated using a nuclear ensemble of 2000 geometries, with vertical excitations calculated at the EOM-CCSD/def2-TZVPD or the wB97X-D/def2-TZVPD levels of theory. The simulated UV-VIS spectra for the smaller species were in quite good agreement with experimental ones. Two different molecules were identified with substantial absorption cross sections in the range of the unknown absorber: The open chain isomer of S3 (3.78×10^-17 cm^2 at 370 nm), and the trigonal isomer of S4 (4.76×10^-17 cm^2 at 360 nm). However, the mixing ratios of these species in the Venusian atmosphere are also needed to make a more conclusive statement. Other polysulfur compounds have insignificant absorption cross sections in the 320-400 nm range and can therefore be excluded. The calculated absorption cross sections can be used to calculate photolysis rates, which can be straight away added to atmospheric models of Venus. In addition, this work will help future space missions to Venus, for example by focusing their search for the unknown absorber.

Surface charging behavior of commercial pure calcite and calcite samples from the Bukov underground research facility was studied by measuring the zeta potential of the samples after equilibration with CO2 gas and determining their cation exchange capacity. For zeta potential measurements, different calcite samples with grain sizes under 25 μm were submerged into five different electrolyte solutions: 0.01 M NaCl, 0.001 M CaCl2, 0.005 M NaCl + 0.0025 M MgCl2, 0.0025 M NaCl + 0.00375 M MgCl2, and a synthetic groundwater simulant (SGW2). A pH range of 6.5 – 11 was established and maintained across sample series for the duration of the experiments, and the samples were equilibrated with CO2 gas by regular bubbling of a 250 ppm CO2/N2 gas mixture into the solutions in 1-hour sessions. The length of the equilibration process varied from three days to one month, and after its conclusion the zeta potential of each sample was measured. The goal of the experiments was to evaluate the feasibility of equilibrating calcite samples with CO2 when studying their zeta potential. Additional goals included comparing different types of calcite in different solutions, as well as determining an optimal duration for equilibration. The results were compared with an earlier study where calcite zeta potential was determined under total CO2 exclusion. For the determination of cation exchange capacity, two cobalt hexamine trichloride solutions were used, one saturated with pure calcite and one saturated with Bukov calcite. Samples of both types of calcite were shaken for 1 hour in their respective solutions, and the ionic composition of the supernatant from each sample was determined using microwave plasma atomic emission spectrometry. Cation exchange capacity was calculated from the measurement results. According to the results obtained, long-term exposure and equilibration with CO2 may have a positive effect on the internal stability of calcite samples and lead to more reliable zeta potential measurements. Samples showed minimal fluctuation in the concentration of Ca in bulk solution after one week of equilibration, and no significant rise or decline in stability was noted to occur if equilibration was continued beyond one week. Compared to CO2 exclusion, equilibration produced more consistent trends in zeta potential values, though the results were heavily dependent on the electrolyte solution used in the experiment. Cation exchange capacity experiments also produced much larger values than the previous exclusion study despite using the same types of sample, hinting at a significant difference in the grain sizes used.

The opening segments give a summary of the history of catalysis in general and of frustrated Lewis pairs in particular, where both intra- and intermolecular types are discussed. In addition, the essences of the mechanisms of action of frustrated Lewis pairs are discussed, covering electric field, electron transfer and radical-type mechanisms. There is also a discussion on the activity of frustrated Lewis pairs towards dehydrogenation reactions, in which lies the main scope of this thesis. The background and intended parameters of the experimental aspects of this work are initially defined in the thesis scope, along with the practical considerations concerning the reagents, equipment, and special conditions for synthetic procedures also detailed here. The intention was to attempt to elucidate the extent of the impact of different factors, in this case solvent, Lewis acidity, Lewis basicity, and irradiation by blue LED, on dehydrogenation of various N-substituted pyrrolidine substrates. The subsequent section describes the synthetic procedures used for attempted syntheses of the amino-borane ligands. Not all syntheses of the desired ligands were successful, but there was enough success and enough pre-generated material to proceed to the next stage. Due to use of blue LED causing various potential radical side reactions, it was thought appropriate to describe the behaviour of both the sample and substrate blanks under irradiation, as well as the intended reactions during the screening process. Screening provided preliminary data on reaction determining factors, such as solvent, Lewis acidity of the catalyst, and substrate structure. Despite the relative difficulty and time-consuming nature of the catalyst syntheses, some interesting new modes of reactivity appear to be accessible, which may be worth investigating more in the future.

The study consists of two parts: examining the accumulation of americium and plutonium to human ribs, and the accumulation to murine osteoblasts. In the cellular studies stable europium was used as an actinide analogue. After removing the organic material from the bones, americium and plutonium were separated from the dissolved bones by using extraction chromatographic methods. DOWEX, U/TEVA, and TRU resins were used to separate plutonium and americium from impurities. The sample activities were measured using alpha spectrometry. On average, 23.03 mBq/kg of plutonium-238, 12.58 mBq/kg of plutonium-239,240 and 9.81 mBq/kg of americium-241 were measured in the bone samples. The activity concentrations are calculated using the wet weight of the bone. Murine preosteobasts (MC3T3-E1) were mineralized for 12 days. The mineral was subsequently collected for SEM/EDS analysis. Non-mineralized cell fractions were collected for a fractioning study, where the cell internalized europium, membrane bound europium and extracellular europium fractions were collected and measured with MP-AES and ICP-MS. The used europium concentrations were below minimum detectable activity of the MP-AES, but measuring the samples on the ICP-MS showed that the percentage of the internalized europium increases at higher europium concentrations in cell culture media. At 0.2 μM europium concentration, on average, 28.6% of the europium was inside the cells and 71.4% in the media, while at 2.0 μM europium concentration, 30.8% of the europium was inside the cells and 68.5% in the media.