The present thesis proposes new methods for obtaining objective and accurate diagnoses in modern healthcare. Non-invasive techniques have been used to examine or diagnose three different medical conditions, namely neuropathy among diabetics, radiotherapy induced erythema (skin redness) among breast cancer patients and diagnoses of cutaneous malignant melanoma. The techniques used were Near-InfraRed spectroscopy (NIR), Multi Frequency Bio Impedance Analysis of whole body (MFBIA-body), Laser Doppler Imaging (LDI) and Digital Colour Photography (DCP). The neuropathy for diabetics was studied in papers I and II. The first study was performed on diabetics and control subjects of both genders. A separation was seen between males and females and therefore the data had to be divided in order to obtain good models. NIR spectroscopy was shown to be a viable technique for measuring neuropathy once the division according to gender was made. The second study on diabetics, where MFBIA-body was added to the analysis, was performed on males exclusively. Principal component analysis showed that healthy reference subjects tend to separate from diabetics. Also, diabetics with severe neuropathy separate from persons less affected. The preliminary study presented in paper III was performed on breast cancer patients in order to investigate if NIR, LDI and DCP were able to detect radiotherapy induced erythema. The promising results in the preliminary study motivated a new and larger study. This study, presented in papers IV and V, intended to investigate the measurement techniques further but also to examine the effect that two different skin lotions, Essex and Aloe vera have on the development of erythema. The Wilcoxon signed rank sum test showed that DCP and NIR could detect erythema, which is developed during one week of radiation treatment. LDI was able to detect erythema developed during two weeks of treatment. None of the techniques could detect any differences between the two lotions regarding the development of erythema. The use of NIR to diagnose cutaneous malignant melanoma is presented as unpublished results in this thesis. This study gave promising but inconclusive results. NIR could be of interest for future development of instrumentation for diagnosis of skin cancer.

Petroleum powered vehicles emit volatile organic compounds (VOCs) through combustion that contributes to the pollution of the environment. A technique in the 1970s was developed to decrease these emissions, especially for nitrogen oxides (NOx) and sulphuric oxides (SOx) which is called exhaust gas recirculation (EGR). The technique works by recirculating a portion of the combusted gas back into the engine, this limits the NOx and SOx emissions because of lower temperatures and less available oxygen. The problems that follow these effects is the formation and condensation of acids that corrode the material of the EGR system, which are created by many different reactions. It is of importance to understand how the compounds in the EGR system behaves through analysis of authentic and simulated condensates, which is why a quantitative method for these compounds are of interest. The aim of the project was to develop a simple quantitative analysis method for formic acid, acetic acid, and lactic acid in aqueous solution, which was done at Gränges Sweden AB. The technique used for detection and quantification was gas chromatography (GC) coupled to a flame ionization detector (FID) and a water compatible polyethylene glycol (PEG) column. Fractional factorial design (FFD) was used for determination of adequate operating parameters of the GC method and the sample preparation. Sample preparation only required filtration and pH adjustment prior to direct aqueous injection (DAI) to the chromatographic instrument. Detection of the analytes was very difficult because of non-compatibility with the FID, and quantification of asymmetric peak shapes made this problem worse, omitting lactic acid from further analysis. Limit of detection (LOD) and limit of quantification (LOQ) was 490 and 1640 ppm for formic acid and 120 and 400 ppm for acetic acid, with an injection volume of 0.3 μL and split ratio 10:1. Limits were too high for every EGR sample leaving no peaks detected for the sample preparation used. Further development should be done with complementary techniques and sample reprocessing in order to quantify the compounds.

Linseed oil has been and is used for vast number of applications, such as in food and paint industry, and wood preservation. It is a good environmental choice, as it originates from renewable sources. Linseed oil is mainly a mixture of triglyceride of fatty acids, both saturated and mono- or polyunsaturated, which allows the oil to oxidize. The oxidation occurs via an auto-oxidation mechanism with the carbon-carbon double bonds and oxygen from the air, reacts to form a polymer. Herein, four different linseed oils (three commercial ones and one industrially available) were analyzed to obtain a better understanding of why different oils provide different protection of wooden materials. This was done by a study of the unoxidized oil, followed by an oxidation time-resolved study of oxidized oil films. The analysis was done by nuclear magnetic resonance, gas chromatography - mass spectrometry and/or inductively coupled plasma atomic emission spectroscopy. This study provided the fatty acid profile of the oils, which were similar for all oils. The unoxidized oils contained some metals ions which probably originate from additives. Aluminum, cobalt, iron, manganese, and zinc was detected in some of the oils at concentrations up to 135 mg/L, but only manganese was detected in all oils and its concentration was much higher than all other metals together. The time-resolved oxidation study had some problems with the solubility of the formed polymers. Several solvents were examined, such as dimethylsulfoxide, alkaline alcohol solutions and toluene, before chloroform-d was chosen as solvent. Though, chloroform-d was not a perfect solvent; it was capable to solve a fraction of the sample, but the fraction decreased with oxidation time. After fifteen days of oxidation, only a few percent of the sample could be dissolved, but for short oxidation times (<48 h) the majority of the samples were dissolved. The oils were analyzed after thirteen different oxidation times. Some structural changes appeared, for example loss of unsaturated protons and some oxidation products arose, such as peroxides and aldyhydes. The diffusion coefficient decreased over the first 3-4 days of oxidation, as expected when the polymerization progressed. After a week of oxidation, the diffusion coefficient increased again, this could possibly be explained by the solubility problem for the large polymer formed. Contrary, at shorter oxidation times this method probably could still be used, as the majority of the sample was dissolved. However, the solubility problem made it impossible to conclude anything about the oxidation rate at longer oxidation times and thus prevented any ranking of the oils. / Linolja har använts och används för många olika tillämpningar, till exempel i mat- och färgindustrin, samt för att bevara trä. Det är klimatmässigt ett bra val, då det kommer från en förnyelsebar källa. Linolja innehåller huvudsakligen en blandning av triglycerider av fettsyror, som både kan vara mättade, enkelomättade eller fleromättade, detta gör att linolja kan oxidera och torka. Oxidationen sker via en auto-oxidation mekanism, med kol-kol dubbelbindningarna och syre från luften som producerar till en polymer. I detta projekt undersöktes fyra olika linoljor (tre kommersiella och en industriell), för att ge en bättre förståelse till varför olika oljor ger olika bra skydd för trämaterial. Detta gjordes genom att undersöka de icke-oxiderade oljorna och sedan göra en tidsstudie på oxiderade oljefilmer. Alla dessa prover analyserades med nuclear magnetic resonance, gas chromatography - mass spectrometry and/or inductively coupled plasma atomic emission spectroscopy. Studien gav resultat på sammansättningen av fettsyror i oljorna, vilken var liknande för alla oljorna. De icke-oxiderade oljorna innehöll ett par metaller, som förmodligen kommer från additiv. Aluminium, kobolt, järn, mangan och zink hittades i några av oljorna i koncentrationer upp till 135 mg/L, men bara mangan var detekterad i alla oljorna och dess koncentration var högre än alla andra metaller tillsammans. Tidsstudien hade problem med lösligheten av proverna. Flertalet lösningsmedel undersöktes, exempelvis dimetylsulfoxid, alkaliska alkohollösningar och toluen, innan kloroform-d valdes som lösningsmedel. Däremot var kloroform-d inte ett perfekt lösningsmedel, den hade förmågan att lösa en del av proverna, men den delen minskade med oxidationstid. Efter femton dagar kunde den bara lösa ett par procent, men efter kortare oxideringstider (<48 h) gick majoriteten av proven att lösa. Oljorna analyserades vid tretton olika oxideringstider. Några strukturella förändringar uppmättes, till exempel minskade mängden dubbelbindningar, och ett par biprodukter från oxideringen detekterades, så som peroxider och aldehyder. Diffusionskonstanterna för oljorna minskade under de första 3-4 dagarna av oxidering, precis som förväntat under polymeriseringen. Efter en vecka av oxidering ökade diffusionskonstanterna igen, det kan förmodligen förklaras av löslighetsproblemen, då diffusionskonstanten är beroende av koncentrationen. Å andra sidan, vid kortare oxideringstider kan denna metod fortfarande användas, eftersom vid denna tidpunkt löste sig fortfarande majoriteten av provet. Dock, löslighetsproblemet gjorde det omöjligt att dra slutsatser kring längre oxideringstider och därmed förhindrades rangordning av oljorna.

Lactate is an important biomarker in sports and the lactate threshold concept is one of the best indicators of endurance performance in an athlete. However, to quantify the lactate threshold, an invasive method to take a blood sample must be used. Limitations for this method include stopping the active exercise for blood sampling and no real-time feedback. Instead, a novel non-invasive wearable biosensor can be used to measure the lactate concentration in sweat. The sensor generates an electrical current depending on the lactate concentration in the sweat, and therefore must an electronic device called a potentiostat be used to condition the current. However, available potentiostats are not suitable for use in sports where form factor, battery life and wireless connectivity is important. This thesis aims to solve this by developing a wearable device and software which can be used to measure the lactate concentration in sweat in real-time during exercise. The development process consisted of the determination of specifications, prototype development and thorough laboratory and on-body testing. Finally, a novel wearable device and software capable of real-time lactate measurements in sweat during exercise were presented. The device, called LactateStat, was 58mm ∗ 55mm ∗ 13mm, the current consumption was only 7.8mA, the current measurement resolution was 0.5 nA, the limit of detection was 0.45 nA and the current measurement range was around 750 μA. LactateStat is one of the first of its kind and provides a base for future development as the hardware, firmware and software resources are provided as open-source. / Laktat är en viktig biomarkör inom idrott och konceptet laktattröskel är en av de bästa indikatorerna för uthållighet hos en idrottsutövare. Men för att mäta laktattröskeln krävs en invasiv metod som går ut på att man tar ett blodprov. Begränsningarna med denna metod är bland annat att idrottsutövningen måste stoppas för att ta ett blodprov och att man inte får realtidsåterkoppling. I stället kan en liten och icke-invasiv bärbar biosensor användas för att mäta laktatkoncentrationen i svett. Sensorn genererar en elektrisk ström beroende på laktatkoncentrationen i svetten och en enhet som kallas potentiostat måste därför användas för att mäta denna ström. Tillgängliga potentiostater är dock inte lämpliga för användning inom sport där formfaktor, batteritid och trådlös kommunikation är viktigt. Denna avhandling har som syfte att lösa detta genom att utveckla en bärbar enhet och mjukvara som kan användas för att mäta laktatkoncentrationen i svett i realtid under träning. Utvecklingsprocessen bestod av bestämning av specifikationer, framtagning av en prototyp och noggrann laboratorie- och kroppstestning. Som resultat presenterades en banbrytande bärbar enhet och mjukvara som kan mäta laktat i realtid i svett under träning. Enheten, som kallades LactateStat, var 58mm * 55mm * 13mm, den aktiva strömförbrukningen var 7.8mA, den bästa mätupplösningen för ström var 0.5 nA, detektionsgränsen var 0.45 nA och det maximala mätområdet för ström var ca. 750 μA. LactateStat är en av de första i sitt slag och ger en bas för framtida utvecklingsarbete eftersom hårdvaru-, programvaru- och mjukvaruresurserna tillhandahålls som öppen källkod.

The lithium-ion battery (LIB) recycling industry is currently under development to improve the yields for critical metals. However, the organic components of LIBs must also be handled, which may result in harmful chemical emissions as by-products. Of particular concern are highly persistent and mobile per- and polyfluoroalkyl substances (PFAS) that could be released during LIB recycling since some of these compounds have been linked with adverse health effects. In this work, an extensive literature review was conducted to determine the presence of fluorinated materials in state-of-the-art LIBs and the recycling conditions which could lead to the release of problematic PFAS. This information was used to develop a new analytical approach to capture the broadest range of organic and inorganic fluorine species in samples taken in different stages of the recycling process. This new method is based on a sequential extraction procedure using different solvents, followed by combustion ion chromatography (CIC) to quantify the potential emission of fluorine-containing chemicals of different polarities. The results show that organofluorine compounds are formed during recycling, particularly for the cathodes, indicating that PFAS might be present. For other samples, such as the NiMnCo salt product of recycling, only low fluorine levels were detected which implies almost complete removal. Future work should further outline the emission paths of these processes. This study highlights the necessity to further investigate the emissions related to fluorinated materials during LIB recycling and indicates that post-treatments or changes in conditions might be necessary to avoid the formation and emission of PFAS.

<p>QC 20110615</p>

The emergence of β-lactamase-mediated antibiotic resistance is one of the biggest threats in modern times. Combined with the discovery void of new forms of antibiotics, this sets the course toward a future where the efficacy of present-day health care will be jeopardized. To hinder the spread of β-lactamase-mediated antibiotic resistance, the development of the drug class β-lactamase inhibitors has been prioritized. The foremost candidate for development of this drug class, that has wide-spectrum inhibition of β-lactamases, is the clinically available avibactam of the diazabicyclooctane-scaffold (i.e., DBO-scaffold). However, the clinical applications of this inhibitor have been limited against one of the more rapidly spreading β-lactamases; OXA-48. In order to bolster the drug development of β-lactamase inhibitors of the DBO-scaffold, with good inhibitory activity toward OXA-48, DBO-ligands with different structure elements were analyzed for stability of the Michaelis Complex in the OXA-48 binding site using molecular dynamic simulations. The results indicate that elongation of the chain to the anionic group of the ligand combined with the addition of a methyl group to the DBO-ring was stabilizing for the productive position between the backbone hydrogens of Y211 and S70. The binding affinity was also estimated using the Linear Interaction Energy method, and an offset parameter of γ ≈ -19 kcal/mol was found and could represent the entropic differences of a flexible ligand-protein system. The results of this study may also indicate that the ligand stability of the Michaelis Complex is of minor consequence to the inhibition mechanism as a whole compared to the reaction rate.

Described in many studies as dangerous for the environment and potentially carcinogenic for humans, Chlorinated Paraffins (CPs) are easily widespread due to their substantial production and use in different products. Previous studies reported the presence of CPs in different plastic polymers. However, the impact of recycled content in plastic materials on the CPs levels hasn’t particularly been considered. Recycling plastics is becoming essential but the accumulative potential of pollutants, such as CPs, need investigations. The presence of CPs in both virgin and mixed recycled and virgin plastics was studied. Plastic pellets and plastic pieces from products made of three polymer types:Poly(methyl 2-methylpropenoate) (PMMA), Thermoplastic Rubber (TPR) and Thermoplastic Polyurethane (TPU) were analysed. A solid-liquid extraction assisted by ultrasonication was performed, followed by cleanup using silica. CPs in plastic extracts were analysed by Gas Chromatography Orbitrap High Resolution Mass Spectroscopy (GC-Orbitrap-HRMS). Because of the high volatility characteristic of long chain Chlorinated Paraffins (LCCPs), only short chain and medium chain Chlorinated Paraffins (SCCPs and MCCPs ) have been studied in this project. Interesting variations in the presence of SCCPs and MCCPs have been observed from one polymer type to another. In the results, we show that MCCPs were less frequently detected compared to SCCPs. A higher detection frequency of CPs was observed for samples containing recycled plastics. The results indicate that CPs are present in plastic polymers (TPU, PMMA and TPR) and that the content of recycled material has a direct impact on the levels of SCCPs and MCCPs.

Diabetic ketoacidosis (DKA) is a life-threatening condition that can appear in patients with diabetes. High ketones in the blood lead to acidity of the blood. For DKA diagnosis and management, ketones such as hydroxybutyrate (HB) can be used to quantify the severity of the disease. The fabrication of electrochemical biosensors for the detection of HB is attractive since their capability to deliver fast response, high sensitivity, good selectivity and potential for miniaturisation. In this thesis, an integrated electrode system was prepared for the detection of HB. Laser-induced graphene (LIG) with a 3D porous structure was used as the flexible platform. Poly (toluidine blue O) (PTB) was electro-deposited on LIG (PTB/LIG) under the optimised conduction (pH of 9.7 and from 0.4 to an upper cyclic potential of 0.8 V). The single PTB/LIG working electrode demonstrated excellent performance towards the detection of NADH with a linear range of 6.7 M to 3 mM using chronoamperometry, high sensitivity of detecting NADH and excellent anti-fouling ability (94 % response current retained after 1500 s). Further integration of the 3-electrode system realised the static amperometric detection of NADH over the range of 78 M to 10 mM. Based on the excellent performance of PTB/LIG to NADH sensing, hydroxybutyrate dehydrogenase was immobilised via encapsulation with chitosan and polyvinyl butyral (PVB) which was used for HB biosensing over the linear range of 0.5 M to 1 mM with NAD+ dissolved in solution. In addition, the co-immobilisation of NAD+ and HBD on PTB/LIG was conducted by optimisation of enzyme and NAD+ amount per electrode, which shows excellent reproducibility and satisfactory HB biosensing performance. Further experiments to improve the long-term stability of the enzyme electrode is expected in the future. The proposed integrated electrode system also possesses the potential to extend to a multichannel sensor array for the detection of multiple biomarkers (e.g. pH and glucose) for diagnosis and management of DKA.

Protein analysis is important to understanding biological systems, but sample diversity necessitates a multitude of analysis techniques and methods. Challenges that are addressed include analysis of low abundance samples, fractionation to reduce sample complexity, and automation to reduce time and cost. Matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) is an important technique for protein characterization. In Paper I, the sensitivity of MALDI-MS was enhanced through the fabrication of a hydrophobic coating for the MALDI target plate, yielding analyte concentration. The plate outperformed a commercial concentration plate. Capillary electrophoresis (CE) separation offers low sample consumption and high efficiency, and in Paper II, offline CE-MALDI-MS fractionation was employed. A robot system for automation was constructed and used in analysis of spermatophore proteins from the butterfly Pieris napi. The robot was also used in automated on-target trypsin digestion under a lid of liquid fluorocarbons, a simpler and cheaper alternative to controlled humidity chambers. An indication of indigenous proteolysis of the sample was seen. Electrospray ionization (ESI) is the other technique for protein analysis in MS. In Paper III, the biomarker protein osteopontin (OPN) was analyzed by ESI-MS in order to find suitable conditions for its detection. A preliminary optimization of solvents and ionization conditions was done, and tandem MS (MSn) performed to increase the reliability of identification. / Proteinanalys är viktigt för att förstå biologiska system, men mångfalden av prov kräver en mängd olika analystekniker och metoder. Utmaningar som tas upp inkluderar analys av små provmängder, fraktionering för att minska provkomplexiteten, och automatisering för att minska tidsåtgång och kostnad. Matris-assisterad laserjoniserings-masspektrometri (MALDI-MS) är en viktig teknik för proteinkarakterisering. I Artikel I förbättrades känsligheten i MALDI-MS genom tillverkning av en hydrofob beläggning på MALDI-provplattan, vilket gav en koncentration av provet. Provplattan gav bättre resultat än en kommersiell koncentrationsprovplatta. Kapillärelektroforesseparation (CE) har låg provåtgång och hög separationseffektivitet och i Artikel II användes offline CE-MALDI-MS-fraktionering. Ett robotsystem för automatisering konstruerades och användes för analys av spermatoforproteiner från fjärilen Pieris napi. Roboten användes även i automatiserad trypsinklyvning under en yta av en flytande fluorkolförening, ett billigare alternativ tilli nkubationskammare med kontrollerad luftfuktighet. En indikation på naturlig enzymatisk proteinklyvning i provet hittades. Elektrospray jonisering (ESI) är den andra tekniken för proteinanalys i MS. I Artikel III analyserades biomarkören osteopontin (OPN) med ESI-MS för att hitta lämpliga förhållanden för dess detektion. En preliminär optimering av lösningsmedel och jonisationsförhållanden gjordes, och tandem-MS (MSn) utfördes för att öka identifikationens tillförlitlighet. / <p>Full text will not be uploaded due to unpublished results. QC 20181121</p>