Phosphorus (P) regulates trophic status in most aquatic systems. However, only bioavailable P contributes to primary production. In most lakes and shallow seas, mineralisation of sediment P into its bioavailable form and its release to the water column is important for maintaining primary production. Sediment organic P forms a substantial proportion of this P to be mineralised and can originate from different sources on land (farmland, forests, etc.) or from primary production in the lake. These organic P forms can thus be expected to have differing composition, degradability and recyclable P content. Knowledge of the chemical structure of sediment organic P compounds is scarce, mainly due to lack of appropriate analytical techniques. The commonly used 31P-nuclear magnetic resonance (31P-NMR) technique, only identifies P binding groups, so a mass spectrometric (MS) analysis method was developed that allows individual sediment organic P compounds to be identified. EDTA as pre-extractant resulted in the highest P yield in subsequent NaOH extraction. Extracted organic P compound groups were identified using 31P-NMR. For identification of specific P compounds with MS, a sample preparation method prior to electrospray tandem mass spectrometry (ESI-MS/MS) analysis was developed. Liquid chromatography (LC) with porous graphitic carbon prior to ESI-MS/MS enhanced sensitivity and selectivity, enabling several of the ions detected to be identified as nucleotides. 31P-NMR analysis showed P monoesters to be the most stabile P compounds throughout a lake sediment profile. The developed LC-ESI-MS/MS analysis method revealed that some monoester-P (nucleotides) were labile, while other P compounds increased in concentration with Baltic Sea sediment depth and were therefore considered stabile. Differences in patterns of P compounds detected were also shown depending on catchment characteristics in relation to Baltic Sea sediment age. For cost-effective management of eutrophication, knowledge of the sources of degradable organic P forms, contributing to internal loading, is needed. This thesis showed the developed LC-ESI-MS/MS analysis method to be a powerful analytical tool for this purpose.

This is to my knowledge the first time a ten-year follow-up study of salivary cortisol concentrations measured by immunoassays in relation to posttraumatic symptoms according to the Impact of Event Scale (IES) is made. The study was performed on 78 Swedish UN soldiers after a 6-months mission in the former republic of Yugoslavia. Follow-up investigations were performed six months, twelve months and ten years after their return to Sweden. Morning and evening salivary cortisol concentrations were determined by radioimmunoassay (RIA) and enzyme-linked immunoassay (EIA) and subjective posttraumatic avoidance and intrusion symptoms were measured with the IES (see Appendix I).   This study concerns the methodological description of the EIA for determination of salivary cortisol and the comparison of the results from all three follow-up investigations. Post-traumatic stress symptoms according to IES (intrusion subscale and total score) increased significantly over ten years of time. There was an significant interrelationship between the change in both morning and evening salivary cortisol concentrations, measured with immunoassays, and changes in self-rated posttraumatic intrusive symptoms, according to IES, during ten years follow-up, after a six months mission in Bosnia in the way that salivary cortisol concentrations showed a tendency to decrease over ten years of time in subjects with a higher IES score. The rise in morning salivary cortisol, from awakening until 30 minutes later, was significantly correlated with the ratings of posttraumatic stress symptoms according to the IES ten years after the mission.

<p><b>In this thesis the fabrication and analytical evaluation of two new electrospray emitters utilized for mass spectrometry analysis is presented. The emitters are based on a new concept, where the spray orifice can be varied in size. The thesis is based on two papers.</b></p><p>All present-day nanoelectrospray emitters have fixed dimensions. The range of the applicable flow rate for such an emitter is therefore rather limited and exchange of emitters may be necessary from one experiment to another. Optimization of the signal of the analyte ions is also limited to adjustments of the applied voltage or the distance between the emitter and the mass spectrometer inlet. Furthermore, clogging can occur in emitters with fixed dimensions of narrow orifice sizes. In this thesis, electrospray emitters with a variable size of the spray orifice are proposed. An open gap between two thin substrates is filled with sample solution via a liquid bridge from a capillary. Electrospray is generated at the end point of the gap, which can be varied in width.</p><p>In Paper I, electrospray emitters fabricated in polyethylene terephthalate have been evaluated. Triangular tips are manually cut from the polymer film. The tips are mounted to form a gap between the edges of the tips. The gap wall surfaces are subjected to a hydrophilic surface treatment to increase the wetting of the gap walls.</p><p>In Paper II, silicon electrospray chips with high precision are fabricated and evaluated. A thin beam, elevated from the bulk silicon chip is fabricated by means of deep reactive ion etching. The top surfaces of the beams of two chips act as a sample conduit when mounted in the electrospray setup. An anisotropic etching step with KOH of the intersecting <100> crystal planes results in a very sharp spray point. The emitters were given a hydrophobic surface treatment except for the hydrophilic gap walls.</p><p>For both emitter designs, the gap width has been adjusted during the experiments without any interruption of the electrospray. For a continuously applied peptide mixture, a shift towards higher charge states and increased signal to noise ratios could be observed when decreasing the gap width. The limit of detection has been investigated and the silicon chips have been interfaced with capillary electrophoresis.</p>

<p>In this final thesis, an existing method for analysis of chlorophenols (CP) in bottom sediments has been updated and adjusted for analysis of chlorophenols in soil. The covalent bonds between the chlorophenols and the soil matrix were broken through basic hydrolysis and the chlorophenols were then separated from the water phase through addition of sulphuric acid followed by ether extraction. The chromatography was improved through extractive acetylation of the chlorophenols.</p><p>The updated method was then applied on soil samples from a contaminated area (a former sawmill in Hyttsjö, Östergötland, Sweden).</p><p>The analyse was preformed by GC/MS with respect to 2-MonoCP, 4-MonoCP, 2,4-DiCP, 2,6-DiCP, 2,4,6-TriCP, 2,3,4,6-TetraCP and pentachlorophenol (PCP).</p><p>Contamination of chlorophenols in nature can be explained by the former use of wood preservative chemicals based on chlorophenols. In the 1960s and the 1970s these chemicals were used in Sweden, but due to their toxicity they were banned by the Swedish government in 1978.</p><p>In Hyttsjö a pentachlorophenol-based product named Santobrite was used for several years. The concentration of PCP in the soil samples from Hyttsjö varied from 0.2->1.8 ng/mg dry substance. 2,3,4,6- Tetrachlorophenol was also detected in some of the soil samples.</p>

High performance liquid chromatography (HPLC) is one of the major techniques in separat-ion sciences. Faster separation and higher efficiency are required to meet ever-growing demands. Despite numerous studies and achievements on improving mass transfer in particulate packings discontinuity seems to be the cornerstone drawback in their development. Macroporous continuous beds or monoliths are therefore a promising alternative to the particle medium. This thesis deals with preparation of new monoliths used as carrier for HPLC. Two different approaches were developed for two polymer systems. One was based on polycondensation of epoxy resins and polyamines which were components of an oil-in-water emulsion. An epoxy resin mixture was dispersed in aqueous polyamine phase with the aid of a surfactant. The other involved a traverse of a ready-made polymer solution around its upper critical solution temperature (UCST). In other words, linear polyamides, non-covalently crosslinked polymers, dissolved in a solvent at temperature higher than their UCST followed by slow cooling to below the critical temperature to precipitate the polymers. Partly re-established hydrogen bonds resulted in the formation of crystallites that interconnected into a network structure. Factors controlling morphology and porosity of final products were investigated. The study also deals with surface modifying for chromatographic applications. Functionalization pathways attempted in the thesis were quaterization of inherent amine of the epoxy-based monoliths and grafting tentacle ion groups via glycidyl methacrylate by atom transfer radical polymerization (ATRP) for ion exchange chromatography (IEC).

The foci of this dissertation are on advanced liquid chromatography (LC) separation and mass spectrometry (MS) techniques for the analysis of small bioactive molecules. In addition to discussing general aspects of such techniques the results from analyses of polyphenols (PPs), alkaloids and amino acids published in five appended studies are presented and discussed. High efficiency and well understood principles make LC the method of choice for separating analytes in many kinds of scientific investigations. Moreover, when LC is coupled to an MS instrument, analytes are separated in two stages: firstly they are separated and pre-concentrated in narrow bands using LC and then separated according to their mass-to-charge (m/z) ratios in the MS instrument. Some MS instruments can provide highly accurate molecular weight measurements and mass resolution allowing identification of unknown compounds based purely on MS data, thus making prior separation unnecessary. However, prior separation is essential for analyzing substances in most complex matrices – especially useful is the ultra-high performance LC (UHPLC). The advantages of using UHPLC rather than HPLC for the analysis of PPs in tea and wine were evaluated in one of the studies this thesis is based upon. The phenolic composition of red wine was also examined, using a novel LDI technique, following solid phase extraction (SPE). A class of small aromatic molecules (medicinally important alkaloids) also proved to be amenable to straightforward analysis, by thin layer chromatography (TLC) work-up followed by LDI-MS. Finally, a LC-MS method for monitoring neurotoxins (β-N-methyl-amino-L-alanine and 2,3-diaminobutyric acid) in complex biological matrices was developed and applied. Overall, the studies show that careful attention to the physicochemical properties of analytes can provide insights that can greatly facilitate the development of alternative methods to analyze them, e.g. by LDI. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 4: In press. Paper 5: Manuscript.</p>

Sample preparation is often a bottleneck in systems for chemical analysis. The aim of this work was to investigate and develop new techniques to address some of the shortcomings of current sample preparation methods. The goal has been to provide full automation, on-line coupling to detection systems, short sample preparation times and high-throughput. In this work a new technique for sample preparation that can be connected on-line to liquid chromatography (LC) and gas chromatography (GC) has been developed. Microextraction in packed syringe (MEPS) is a new solid-phase extraction (SPE) technique that is miniaturized and can be fully automated. In MEPS approximately 1 mg of sorbent material is inserted into a gas tight syringe (100-250 μL) as a plug. Sample preparation takes place on the packed bed. Evaluation of the technique was done by the determination of local anaesthetics in human plasma samples using MEPS on-line with LC and tandem mass spectrometry (MS-MS). MEPS connected to an autosampler was fully automated and clean-up of the samples took about one minute. In addition, in the case of plasma samples the same plug of sorbent could be used for about 100 extractions before it was discarded. A further aim of this work was to increase sample preparation throughput. To do that disposable pipette tips were packed with a plug of porous polymer monoliths as sample adsorbent and were then used in connection with 96-well plates and LC-MS-MS. The evaluation of the methods was done by the analysis of local anaesthetics lidocaine and ropivacaine, and anti-cancer drug roscovitine in plasma samples. When roscovitine and lidocaine in human plasma and water samples were used as model substances, a 96-plate was handled in about two minutes. Further, disposable pipette tips may be produced at low cost and because they are used only once, carry-over is eliminated.

Semiochemicals are chemicals used to communicate. Animals tend to use these e.g. to locate food sources or to find a suitable mate. In this study, the sex pheromone of the Indian meal moth, Plodia interpunctella, was analysed. Since this is an economically important species, it is mass-reared in labs and science centers worldwide for experimental purposes. A culture of these moths was brought into the lab at Lund University for studies and has after that served as a model species demonstrating up-wind pheromone-mediated male flight in different courses held by the university. As years went by, the culture got less successful in up-wind flights, most probably because of inbreeding and bottleneck effects, and therefore, a new culture was taken in. This study focuses on using various experiments to see if there was a behavioral and/or physiological difference between the two cultures. Results show a significant difference in behavioral traits (frequency of calling behavior in females and in male up-wind flights) but not in physiological traits (female pheromone production or male antennal response). This study discusses some effects of mass-reared lab cultures.

The hip-flask defence is a common claim in drunkdrinking cases. In Sweden and Norway two different models are used to determinethese cases. In Sweden one blood and two urine samples taken 60 minutes apartare used for analysis. In Norway two blood samples taken 30 minutes apart areused. Sweden focuses on the rise or fall of alcohol concentration in urine(UAC), and the ratio between UAC and blood alcohol concentrations (BAC). Norwayfocuses on the rise or fall of the alcohol metabolite ethylglucuronide (EtG) and the ratio between BAC and EtG. The aim of this study wasto test the models for multiple intakes and with different alcoholic beverages.Thirtyfive participants ingested two doses, first0.51 g/kg of beer and later either 0.25, 0.51 or 0.85 g/kg of beer, wine orvodka. Blood and urine samples were obtained before and after alcoholingestion. Alcohol was measured by GC-HS, and the alcohol metabolite byUPLC-MS/MS. The results showed that there are kineticdifferences between single and repeated intakes, that there are no significantdifferences in kinetics from different alcoholic beverages and thatthe Norwegian model appears to be the stronger one in hip-flask determination.

As of today, synthetic cannabinoids are one of the biggest groups of new psychoactive substances.These substances can be used as substitutes for the psychoactive drug cannabis, avoiding the legalrestrictions on cannabis. Furthermore, a variety of synthetic cannabinoids are synthesized with eithersignificant or very minor structural differences, making the detection of said novel drugs hard to keepup with and is therefore of great importance to have standards which help in the identification of theintake of the parent synthetic cannabinoid. In this project, several metabolites ofsynthetic cannabinoids with indole/indazole cores with differentside chains was synthesized. The general strategy used in this project was to N-alkylate the desiredcore followed by amide coupling with L-tert-leucine methyl ester or L-Valine methyl esterhydrochloride which resulted in 8 potential synthetic cannabinoid metabolites.