Syftet med denna studie var att utveckla en separations- och analysmetod för identifiering av icke-volatila substanser från strån och rötter av de gräsliknande växterna Cyperus rotundus och Panicum repens. För analysen användes högpresterande vätskekromatografi (HPLC) och ultrahögpresterande vätskekromatografi (UHPLC) med en opolär stationär fas (RP-HPLC) med ultraviolett ljus (UV) och elektrosprej masspektrometri detektion (ESI-MS). Proverna extraherandes med hjälp av ultraljuds assisterad extraktion (UAE) och koncentrerades med solid fasextraktion. Två olika metoder användes vid extraktionen antingen sänktes gräset i 10 ml acetonitril två gånger eller så direkt i 20 ml acetonitril. På grund av svårigheten att uppnå reproducerbarhet vid extraktion med samma metod kunde ingen slutsats om någon metods fördelar dras. Under utvecklingen av HPLC metoden testades två olika elueringsmedel metanol och acetonitril i kombination med MilliQ-vatten. Den slutgiltiga metoden börjar med en mobilfas innehållande 70% metanol och 30% vatten under 5 min för att sedan öka till 100% metanol under 10 minuter innan mobilfasen hålls vid 100% metanol under 25 minuter. Med UV-detektorn kräver identifikation av de okända komponenterna krävde kända referenser. Vidare undersökningar genomfördes därför med masspektrometri. Dock så var det begränsad tid kvar för undersökningar med MS så få körningar kunde genomföras, därav kunde inte analyterna analyseras tillräckligt för att kunna identifiera komponenterna från Cyperus rotundus. Vidare analyser med MS samt även kopplad MSMS där spektrum kan jämföras med en databas för identifikation skulle underlätta identifikationen. / The aim of this study is to develop an analysis method for the separation and identification of non-volatile substances from the Cyperus rotundus and- Panicum repens roots and shoots. For the analysis reversed phase high performance liquid chromatography (HPLC) and ultra-high performance liquid chromatography (UHPLC) methods where used. First an UV detector was used in combination with the HPLC setup and in the later stages access to an electrospray ionization ion trap mass spectrometer was enabled for coupling to UHPLC. The samples were extracted with ultrasound assisted extraction (UAE) and the concentrated with solid phase extraction (SPE). Two different extraction methods were attempted one where the graminoid plants where submerged in 10 ml acetonitrile twice and the other with 20 ml directly. Due to the difficulty of reproducing result between different extraction batches no conclusion on whether either method is superior could be made. For the development of the HPLC method methanol or acetonitrile was used in combination with ultrapure (MilliQ) water. The final eluationprogram was 70% methanol and 30% MilliQ water for five minutes, increasing linearly over 10 minutes to 100% methanol and then running for 25 minutes. With the UV detector no identification was possible, so further investigations were performed with UHPLC-MS for analysis of the mass of the analytes. Due to time limitations only a few MS analyses on the Cyperus rotundus were performed, and only a few masses could be estimated. Further runs of the samples need to be performed in combination with coupled MSMS to be able to identify the analytes in the samples.

I den här studien har metoder för att separera immunoglobulin (IgG) utvecklats med hjälp av kapillärelektrofores (CE) och Ultra högupplösande vätskekromatografi-Mass spektrometri (UHPLC-MS). Vid analysen av peptider från IgG spjälkat av trypsin (IgG TD) detekterades 26 glykopepeptider med UHPLC-MS genom att använda olika gradienter och mobilfaser. Dessutom analyserades fyra olika koncentrations av IgG TD för att testa detektionsgränsen. Resultatet visade att glykopeptider kunde detekteras i alla fyra koncentrationer men antalet detekterade glykopeptider minskade med minskande koncentration. Den sista delen av UHPLC-MS analysen var en selektivitetsanalys där en blandning av bovint serumalbumin spjälkat av trypsin (BSA TD) och IgG TD analyserades i olika koncentrationer. Selektivitetsanalysen visade att ungefär samma antal IgG glykopeptider detekterades som vid analys av IgG TD. Vid CE-analysen användes tre olika kapillärer, varav två av dem var polyvinylalkohol-coated och olika buffertlösningar (BGE) användes. De slutgiltiga resultaten visade 5 toppar som skulle kunna representera IgG gykopeptider men eftersom ingen MS utfördes kunde inga slutsatser dras. Slutligen, användes Matrix-Assisted Laser desorption/ionisation-time-of-flight (MALDI-TOF) för att analysera proverna innan analys och för att jämföra resultaten med UHPLC-MS. / In this study, methods to separate immunoglobulin G (IgG) glycopeptides were developed using capillary electrophoresis (CE) and ultra-high-performance liquid chromatography-Mass spectrometry (UHPLC-MS). In the UHPLC-MS- analysis, 26 glycopeptides were detected during the analysis of trypsin digested IgG (IgG TD) using different gradients and mobile phases. Moreover, a limit of detection study with four different concentrations (0.01, 0.1, 0.5 and 2mg/mL) was performed. Although the number of detected glycopeptides decreased with decreasing concentration, glycopeptides were detected in the 0.01 mg/mL IgG TD sample. Lastly, a selectivity study was performed, in which two trypsin digested Bovine serum albumin (BSA TD) and IgG TD mixtures with different concentrations were analysed. The results showed that approximately the same number glycopeptides were detected as in the IgG sample suggesting that the BSA sample did not interfere with the detection of glycopeptides. Moreover, the CE-analysis included three different capillaries and several background electrolytes (BGEs). Two of the capillaries were polyvinyl alcohol (PVA) coated. The final electropherogram showed five peaks possibly representing IgG glycopeptides but no MS were performed on the CE part leading to no conclusions for the CE-part. Matrix-Assisted Laser Desorption/Ionisation- time-of flight (MALDI-TOF) was performed throughout the project to control the samples and for comparison to the UHPLC-MS analysis.

Huvudmålet i projektet var att analysera det vanligt förekommande additivet glycerolmonostearat, GMS, i plastprover. Projektet är en del av ett samarbete mellan KTH och det norska företaget Norner. Projektgruppen har jobbat för att utvärdera och optimera företagets nuvarande analysmetod samt att identifiera glycerolmonostearat i polypropylen. Olika lösningsmedel och derivatiseringsreagens testades, såväl som olika temperaturprofiler, för gaskromatografen jämfördes under projektet. Resultaten visade att glycerolmonostearat kunde derivatiseras genom silylering. Alla prover har sammanfattats i en slutgiltig metod som är lämpad för analys av polyolefiner som innehåller glycerolmonostearat. Slutmetoden saknade möjligheten för kvantitativ analys och kan därför förbättras i framtiden. / The main task was to analyze the very common additive, glycerol monostearate, GMS in plastic samples. The project was a collaboration between KTH students and the Norwegian company Norner. The goal was to evaluate and optimize the company's current method of analysis and identification of the content of GMS in polyolefins. Different solvents, derivatization agents and methods for gas chromatography were compared to find an effective process by which additives in polypropylene samples can be analyzed. The results showed that glycerol monostearate can be derivatized through a process called silylation. All different attempts have been summarized into one final method that was most suitable for the analysis of polyolefins containing glycerol monostearate. The final method was reproducible but lacked a properly determined quantification analysis. The final method can be improved further after this project. To give a few examples there are a lot more potential derivatization agents and solvents that can be substituted which requires further research.

Klimatförändringarnas påverkan på östra Mälaren undersöktes för att bättre kunna förstå framtida risker för dricksvattenproduktionen i Görväln. Genom att analysera tidigare forskning och vattenkvalitetsdata mellan åren 1968 och 2024, identifierades olika trender med avseende på bland annat temperatur, istäckta dagar, syrgashalt, näringsämnen, växtplankton och olika miljögifter. Utmaningarna är många där varmare klimat kan leda till bland annat tidigare skiktning, syrebrist, isfria vintrar och tillväxt av växtplankton. Även en prognosticerad befolkningsökning runt Mälaren kan bland annat leda till ökande mängd miljöföroreningar och övergödning på höst och vår. Klimatförändringarna förväntas öka yttemperaturen i Görväln men dess påverkan på temperaturen i djupet är inte lika tydlig. En starkare skiktning har observerats, vilket kan leda till låga syrgashalter vid botten, men på grund av Görvälns korta omsättningstid (0,4 år) kan det tänkas att dess påverkan på vattenkvaliteten är marginell. Växtplanktonbiomassan har varierat över tid i Görväln, med en minskning mellan 1968 och 1997 följt av en ökning, som sammanfaller med en höjd vattenmedeltemperatur. Generellt är biomassan högre i Ekoln än i Görväln och Prästfjärden, men även cyanobakteriermängden och totalfosfor är högre för Ekoln. Miljöföroreningarna arsenik, bly och benso(a)pyren visar nästan inga signifikanta trender på grund av begränsad data, men halterna borde ha minskat enligt nationella trender. Framtidens klimatförändringar har påverkat och kan komma att påverka Mälarens vattenkvalitet och därmed utmana dricksvattenreningen för befolkningen runt Mälaren.

Sökhundar är ett viktigt verktyg som används av polis och tullverksamhet, men deras detektionsmetodik är inte helt känd. En substans doftprofil är en viktig aspekt i sökhundars detektion, men de är svåra att forska på kemiskt då det finns en diskrepans mellan koncentration och bidrag till doftprofilen. Målet med den här forskningen var att öka förståelsen för sökhundars detektionsmetodik i relation till adsorberade föreningar på damm, samt hur marijuanas doftprofil skiljer sig från doftprofilen av damm som kontaminerats med marijuana. Citron och lime utnyttjades som modellsystem och bitar av deras skal analyserades även direkt. Terpener och terpenoider var de främsta komponenterna av intresse. Forskningen hade ett tvåfaldigt tillvägagångssätt och inkluderade termisk desorption gaskromatografi masspektrometri (TD-GC/MS) samt sökhundsanalys i form av urvalsövningar. Kontorsdamm och köpt standarddamm i storleken av luftburna partiklar (PM) med en aerodynamisk diameter av cirka 10 µm kontaminerades med marijuana, citron, lime och en cannabis-terpenmix i exsickatorer i 1, 3, samt 7 dagar. En sekundär kontaminering med marijuana utfördes också, där dammprovet som kontaminerats med marijuana i 1 dag användes för att vidare kontaminera nya damm- och PM-prover, och dammprov togs även i förvaringsskåp för packade illegala droger. Från TD-GC/MS analysen av laborativ kontaminering av marijuana detekterades totalt 25 föreningar av intresse i dammprovet, och 57 i PM-provet. Från TD-GC/MS analysen av citron framkom det att fler monoterpener (C10H18) än sesquiterpener (C15H24) detekterades i direktanalysen av citronskal, och det motsatta i analysen av de kontaminerade damm och PM-proverna. Utgående från detta är det troligt att doftprofilerna från ren marijuana samt marijuana kontaminerad på damm eller PM också skiljer sig åt. Sökhundarna markerade dock på båda proverna som kontaminerats med marijuana under 1 dag med stor säkerhet, och vissa markerade även på prover med kortare kontamineringstid och proverna från förvaringsskåpet. Inga eller få föreningar av intresse detekterades med TD-GC/MS i vissa av de prover som hundarna markerade på, vilket exemplifierar att hundarna har en högre sensitivitet vilket försvårar forskningen kring doftprofiler som hundarna kan detektera. Användningen av sökhundsanalys på dammprover som tagits från platser av intresse kan vara en möjlig metod för att detektera marijuana, och eventuellt även andra illegala substanser. / Detection dogs are an important tool utilized by police forces and customs, but their method of detection is not fully known. The odor profile of a substance is of major importance during dog detection, but they are difficult to research chemically due to the disconnection between concentration and contribution to the odor profile. The purpose of this research was to increase the knowledge regarding the mechanism of detection dogs in relation to adsorbed components on dust, as well as of how the odor profile of marijuana differs to that of dust contaminated with marijuana. Lemon and lime were utilized as model systems and pieces of peel were analyzed directly. Terpenes and terpenoids were the main compounds of interest. The research consisted of a dual approach, utilizing both thermal desorption gas chromatography mass spectrometry (TD-GC/MS), and detection dog analysis in the form of selection exercises. Office dust and bought particulate matter (PM)-like standard dust with an aerodynamic diameter of about 10 µm was contaminated with marijuana, lemon, lime, and a cannabis terpene mix in desiccators for 1, 3, and 7 days. A secondary marijuana contamination was also conducted, where the 1-day dust sample contaminated with marijuana was used to contaminate new dust and PM samples, and dust samples were also taken in storage units of packaged illicit drugs. From the laboratory contaminated samples with marijuana, a total of 25 compounds of interest were detected by TD-GC/MS from the dust samples, and 57 from the PM samples. From the direct TD-GC/MS analysis of lemon peel, it was evident that more monoterpenes (C10H18) than sesquiterpenes (C15H24) were detected, and the contrary was found for the lemon contaminated dust and PM samples. This entails that the odor profiles of marijuana as is and adsorbed on dust or PM is also likely to differ. Still, the detection dogs were able to detect the dust and PM samples which had been co-stored with marijuana for 1 day, and some also alerted to a few of the secondary contaminated marijuana samples as well as the samples from marijuana storages. None or only a few compounds of interest were detected by TD-GC/MS in some of the samples the detection dogs alerted to, which highlights the higher sensitivity of detection dogs, and thus the difficulty of chemically researching odor profiles the dogs can detect. All in all, detection dog analysis of dust collected at scenes of interest may be a viable method to detect marijuana, and perhaps other illicit drugs.

Topical inquiries for the Swedish Defence Research Agency (FOI) include analysis of explosive substances in different sample types. Research into explosives in complex matrixes can provide an analytical support function for forensic investigation i.e. tools for areas such as finding bomb factories, identification and risk analysis of home-made explosives (HME) and improvised explosive devices (IED) as well as preventive measures against maliciously intended use of explosives. Additionally, the research may lay the groundwork for indications of health- and environmental hazards. Utilizing state-of-the-art equipment and years of extensive expertise, FOI is able to carry out these types of research tasks to provide security and sustainability for society. The aim of this thesis project is to establish and validate developed methods for collecting, extracting, separating, and detecting trace amounts of explosives in various bodies of water using a solid-phase extraction (SPE) robot and a high-resolution (HR) mass spectrometer (MS) connected to an ultra-high-performance liquid chromatograph (UHPLC). Particular areas of interest include locations in the Stockholm archipelago where experimental detonations of explosives have taken place. Overall, UHPLC-HRMS analysis provides a powerful tool for analyzing explosives in complex matrixes with unambiguous and reliable measurement data. The compounds of investigation were hexogen (RDX), octogen (HMX), pentyl (PETN), and trotyl (TNT). To summarize, during the course of the thesis, trace amounts of some explosives were detected and quantified in various bodies of water. Furthermore, the applied method for the project was successful in qualitatively and quantitatively analyze the compounds of interest with limit of detection ranging between 0.33–11 μg/L (ppb) in various water sources.

<p>Disease-specific compounds (biomarkers) are analyzed in clinical laboratories to assist with diagnosing diseases. This thesis describes development and validation of liquid chromatography tandem mass spectrometry (LC-MS/MS) based tests for diagnosing a diverse group of endocrine and metabolic diseases. The analytical methods used on-line and off-line sample extraction and analytical derivatization as means of enhancing the analytical sensitivity, specificity and clinical utility. All developed methods were extensively validated and reference intervals for the biomarker concentrations were established in blood samples of healthy adults and children. Advantages of the LC-MS/MS as an analytical technique include possibility of simultaneous measurement of multiple analytes and ability of confirming their identity. In this thesis we proposed and evaluated approaches for the assessment of the specificity of analysis in the methods that use tandem mass spectrometry detection. To enhance throughput of the LC-MS/MS tests for the biomarkers that have endogenous or exogenous isomers an approach was developed for quantitation of isomers from unresolved chromatographic peaks. Using methods developed in this thesis we performed a study of the steroidogenesis in ovarian follicles of healthy women and women with polycystic ovary syndrome (PCOS). Obtained data on the steroid concentrations and associations between the steroid metabolites in the pathway would be helpful for better understanding of the ovarian pathophysiology. Potential biomarkers of PCOS were identified in the thesis; further studies will be necessary to confirm their clinical utility.</p>

Until now, most metabolomics protocols have been optimized towards high sample throughput and high metabolite coverage, parameters considered to be highly important for identifying influenced biological pathways and to generate as many potential biomarkers as possible. From an analytical point of view this can be troubling, as neither sample throughput nor the number of signals relates to actual quality of the detected signals/metabolites. However, a method’s selectivity for a specific signal/metabolite is often closely associated to the quality of that signal, yet this is a parameter often neglected in metabolomics. This thesis demonstrates the importance of considering selectivity when developing NMR and LC-MS metabolomics methods, and introduces a novel approach for measuring chromatographic and signal selectivity in LC-MS metabolomics. Selectivity for various sample preparations and HILIC stationary phases was compared. The choice of sample preparation affected the selectivity in both NMR and LC-MS. For the stationary phases, selectivity differences related primarily to retention differences of unwanted matrix components, e.g. inorganic salts or glycerophospholipids. Metabolites co-eluting with these matrix components often showed an incorrect quantitative signal, due to an influenced ionization efficiency and/or adduct formation. A novel approach for measuring selectivity in LC-MS metabolomics has been introduced. By dividing the intensity of each feature (a unique mass at a specific retention time) with the total intensity of the co-eluting features, a ratio representing the combined chromatographic (amount of co-elution) and signal (e.g. in-source fragmentation) selectivity is acquired. The calculated co-feature ratios have successfully been used to compare the selectivity of sample preparations and HILIC stationary phases. In conclusion, standard approaches in metabolomics research might be unwise, as each metabolomics investigation is often unique.  The methods used should be adapted for the research question at hand, primarily based on any key metabolites, as well as the type of sample to be analyzed. Increased selectivity, through proper choice of analytical methods, may reduce the risks of matrix-associated effects and thereby reduce the false positive and false negative discovery rate of any metabolomics investigation.

The revision of the CEN/TR 15522-2:2012 methodology contains new biomarkers to facilitate forensic fingerprinting of the variety of oil types that can be a part of different crimes and the purpose of this project is to validate the biomarkers of the new methodology. Biomarkers were validated by examining corresponding diagnostic ratios compatibility with the internationally used sampling cloth, discriminating power, correlation and simulated weathering sensibility through GC-SIM-MS analysis followed by statistical evaluation with t-tests, diagnostic power, Pearson correlation matrices and MS-PW plots respectively. Results based on most of the diagnostic ratios showed good compatibility with the internationally used sampling cloth, expected patterns of biodegradation and photo-oxidation except for observed photo-oxidation of hydro PAHs and that normative ratios and informative ratios with high diagnostic powers, but with strong correlations for some of the tested ratios, could be identified in diesel oils. Due to delimitations however such as the limited number of oils with similar origins that were analyzed the results should be regarded as guidelines that can be expanded.

The effects of microplastics on zooplankton are an increasing environmental concern. Both primary microplastics that are produced as constituents of cosmetic products, as well as secondary microplastics that are formed by degradation of larger plastic debris, are ubiquitous in aquatic environments. Today, primary microplastics are being phased out and replaced by plant-derived bioplastics. Whether these new materials have similar effects as oil-based microplastics on animals is currently unknown. Here, we compare the effects of secondary microplastic exposure to Daphnia magna, using polylactic acid (PLA) as a representative for bioplastics and polystyrene (PS) for oil-based plastics. To increase the ecological relevance of our tests, we also provided treatments where the particles were coated with bovine serum albumin (BSA) as a means to simulate the coating of biofilms which readily form on particles under natural conditions. Furthermore, to be able to differentiate the effects of general particles from those specific to microplastics, kaolin clay was used as a control treatments, as well as one treatment containing only algae. The objectives were to test the influence of particles on feeding rates, reproduction and growth. PS caused a higher mortality, decreased feeding rate and reproductive output, while PLA and kaolin did not produce any negative effects. BSA did not have a significant effect on reproduction or growth. However, a decrease in reproduction was observed in the plastic treatments. Degradation of PS into styrene monomers is suggested as a possible explanation for the observed toxicity and effects on life history parameters.