In the biopharmaceutical industry, identification of glycans in a glycoprotein is a regulatory requirement and is a part of the characterization of the protein. Glycans are constructed of several monosaccharides linked together. N-linked glycans, which have been studied in this project, are attached to the nitrogen atom in asparagine. A method for separating N-linked glycans by high performance anion exchange chromatography had already been developed at the department. To develop a method for identification of the N-glycans by mass spectrometry, a desalting method on porous graphitic carbon (PGC) columns was used and optimized resulting in the eluents A (0,05% TFA in ACN:water 5:95 v/v) and B (0,05% TFA in ACN:water 50:50 v/v). Also the sample introduction on the mass spectrometer was optimized and resulted in a sensitive on-line liquid chromatography mass spectrometry (LC-MS) approach which gave mass spectrometric peaks with high signal to noise ratios and with high mass accuracy. The developed procedure was then successfully used on glycans cleaved from a glycoprotein separated by high performance anion exchange chromatography with pulsed amperometric detector.

The main purpose of the project was to identify potential environmentally harmful substances which can be leached from rubber asphalt. A method for analysing asphalt was developed and three rubber asphalt materials were analysed after being cryogrinded. One of the materials was also tested in a road machine made for testing of asphalt paving. The particles created in the machine were analysed in the same manner as the cryogrinded asphalt materials. The asphalt materials were leached by water during 24 hours. The leachates were extracted with dichloromethane, dried with sodium sulphate and concentrated to a small volume. The extracts were analysed with gas chromatography-mass spectrometry (GC-MS). Due to low concentrations of substances the GC-MS was operated in SIM-mode (Selected Ion Monitoring). Thirteen substances were chosen for the analysis. The substances were aniline, benzothiazole, butyl benzyl phthalate, bisphenol-A, decanoic acid, dibutyl phthalate, diethylhexyl phthalat, phenanthrene, chrysene, naphthalene, 4-n-nonyl phenol and 4-tert-octyl phenol. Benzothiazole and 4-tert-octyl phenol were detected and quantified in the leachates. In addition to the analysis of organic substances, pH was measured too. The leachates produced in this project were also sent to an analysis company for several analyses, for example analysis of metals and sulphur. Toxicity tests were performed on the same leachates within an exam work made by Gro Runeman at Lund University. The results of the metal analysis, sulphur analysis, and toxicity tests are not covered within the scope of this report. For the results of these tests and analyses, see Gro Runeman’s report: Evaluating toxicity of asphalt leachates. / Detta examensarbete är utfört vid Statens Geotekniska Institut (SGI). Huvudsyftet med arbetet är att identifiera potentiellt miljöfarliga ämnen som kan laka ut från gummiasfalt. En metod för analys av asfalt har tagits fram och tre olika gummiasfalter har analyserats efter att ha kryomalts. En av dessa asfalter har också använts i en provvägmaskin där det damm som bildades har samlats upp och analyserats. Inom ramen för examensarbetet har också en litteraturstudie gjorts för att bland annat ta reda på vad som är gjort inom området sedan tidigare. För att ta reda på vad asfalten innehåller för föreningar gjordes först en fastfasextraktion av krossad (ej kryomald) asfalt där diklormetan användes som lösningsmedel. Från början var tanken att en GC/MS-screening skulle göras för att på så sätt få en överblick av samtliga organiska ämnen som finns i asfalten men på grund av de väldigt låga halterna av i gaskromatografen analyserbara föreningar var det nödvändigt att begränsa analysen till några få föreningar. De föreningar som analysen inriktade sig mot var anilin, bensotiazol, bensylbutylftalat, bisfenol-A, dekansyra, dibutylftalat, di(etylhexyl)ftalat, fenantren, krysen, naftalen, 4-n-nonylfenol, pentaklortiofenol och 4-tert-oktylfenol. Laktester utfördes genom att de kryomalda asfaltmaterialen lakades med vatten under 24 timmar. Efter extraktion, torkning och koncentrering analyserades lakextrakten med avseende på de föreningar som hittats vid fastfasextraktionen. I lakvattnen hittades bensotiazol samt 4-tert-oktylfenol. Dessa föreningar kvantifierades genom att en enpunktskalibrering gjordes. Utöver analyserna ovan mättes även pH på lakvattnen. Lakvattnen skickades också på metall- och svavelanalys samt turbiditets- och fenolindexmätning till ett större analysföretag. Toxicitetstester har utförts på samma lakvatten av Gro Runeman inom ramen för ett examensarbete vid Lunds Universitet. För resultaten från toxicitetstester, metallanalyser samt turbiditets- och fenolindexmätningar hänvisas till Gro Runemans rapport: Evaluating toxicity of asphalt leachates. Delar av denna rapport är skrivna i samarbete med Gro Runeman.

Cystatin C, a 13 kDa low molecular weight protein is an inhibitor of cysteine proteases. Due to its low molecular weight and positive charge at physiological pH, it is freely filtered by the glomerulus and catabolized after reabsorption by proximal tubular cells with a low concentration (0.03-0.3 mg/L) in urine amongst healthy subjects. Urinary cystatin C is a potential biomarker detection of acute kidney injury (AKI) in the acute phase when patients are submitted to the intensive care unit. The aim in this report was to perform a full method validation of urinary analysis of cystatin C on a high throughput chemical analyzer by particle-enhanced turbidimetric immunoassay (PETIA) at the University Hospital in Uppsala, Sweden. The antigen excess, linearity, lower limit of quantification (LoQ), recovery, assay precision, stability and interference caused by haemoglobin was evaluated. No hook effect was observed, the assay was linear over the studied interval <0.001-0.950 mg/L with a regression of R2=0.9994. The LoQ was calculated to 0.020 mg/L with a coefficient of variation (CV) ≤10% which was considered acceptable. The assay had a recovery between 93-100% and the assay precision had a total CV <3.5%. Cystatin C is stable for 3 days in room temperature and 14 days in +4C. The assay did not show any major interference with haemoglobin. The urinary cystatin C showed good precision and performance characteristics by measurements using PETIA all of which is a necessary qualification for a biomarker at a 24-h running routine laboratory.

Syftet med utvecklingsarbetet var att skapa ett genusneutralt läromedeli ämnet kemi om atomen för årskurserna 4-6. Nuvarande läromedelupplevs som för svåra och irrelevanta för eleverna som i stor gradtappar intresset i elvaårsåldern. Framför allt flickor upplever att ämnetinte är riktat till dem. Därför skapade jag ett material utan traditionelltmanliga inslag, för att inkludera alla elever. Fakta förenklades ochkonkretiserades så att den blev relevant och begriplig för eleverna.Produkten syftade till att vara åldersanpassad och kreativtstimulerande, då barns kunskaper gynnas av lekfullt lärande. Materialetbestod av texter och övningar med tillhörande praktiskt material.Produkten testades av åtta flickor och sju pojkar i årskurs fem. Åsikteroch synpunkter om produkten samlades in via en enkät. Enligtenkäterna skapade mitt material nyfikenhet för ämnet, både blandflickor och pojkar. De upplevde att de tittade och läste i boken för att detvar underhållande, inte för att de skulle lära sig något som står ikursplanen. Metoden att skapa ett annorlunda läromedel är ett enkeltknep för att tillvarata det naturliga intresset eleverna har.

Haptoglobin is an acute phase protein with important biological role because of its capacity to bind to haemoglobin. Haptoglobin exists in three major genetic polymorphism types: Hp1-1, Hp2-1 and Hp2-2, the distribution of which has been associated with abdominal aortic aneurysm (AAA), an asymptomatic aortic disease common among men older than 65 years.    Five different electrophoretic methods were tested according to their ability to separate the haptoglobin phenotypes. The detection was based on a produced hemolysate of blood in which haemoglobin binds to haptoglobin thereby forming a complex that can be detected by specific haemoglobin staining using TMB-dihydrochloride and hydro peroxide as substrate resulting in an azure-green color of the bands. Samples from 15 patients who had suffered surgery for not broken AAA, that is more than5.0 cmaortic diameter, and 15 samples from matched controls were analyzed.    Among the five tested electrophoretic methods best migration and separation was seen on the pre-cast agarosgel Hydragel HR on the instrument Hydrasys. The other four methods gave less successful results. This pilot investigation showed the following distribution of the phenotypes of haptoglobin among AAA patients; 7 % Hp1-1, 40 % Hp2-1 and 53 % Hp2-2 and for the controls; 13 % Hp1-1, 33 % Hp2-1 and 53 % Hp2-2.    In conclusion, the used techniques has to be further optimized and more patients have to be included in the study before it can be ascertained if the phenotypes of Haptoglobin play any role in the progress of the AAA disease.

A new method for analysis of acidic herbicides, mostly phenoxy acids and their esters, in cereals with liquid chromatography-tandem quadrupole mass spectrometry (LS-MS/MS) has been developed. Samples were hydrolyzed with sodium hydroxide in order to release covalently bound compounds followed by neutralization and finally extraction with acidified ethyl acetate. The extraction efficiency for both ester formulations and acids were studied. Acceptable results (70-120 %) were obtained for 2,4-D, dichlorprop, MCPA and mecoprop for both esters and acids. However, low recoveries were observed for ester formulations of dicamba, fluroxypyr, fluazifop and haloxyfop, possibly due to the complex structure of the compounds in combination with the matrix and/or incomplete hydrolysis step. The limit of quantification (LOQ) for targeted pesticides was 0.01 mg/kg. The method has been tested in the EU Proficiency Test for cereals with good results.

Correlation between production method and mineral-, cadmium- and protein content were sought for Swedish winter wheat. The wheat was grown according to conventional, organic or biodynamic principles. The minerals; iron, zinc, magnesium, copper and the heavy metal cadmium were analyzed with atomic absorption spectroscopy. Protein was analyzed with elemental combustion analysis. All together, 17 samples were analyzed; nine from the still ongoing Bollerup field trial in Skåne and eight from four different farm pairs in the midst of Sweden. The Bollerup field trials showed that organic wheat had a higher concentration of iron than conventional. Both organic and biodynamic wheat had a higher concentration of zinc than conventional. Magnesium concentration was higher in biodynamic wheat than in conventional grown wheat. Differences between production methods were in the range of 7.5 and 17%. No unmistakable connections were found between any of the parameters and production method for the farm pairs. However, a tendency for more minerals in organic/biodynamic wheat was seen for all minerals but iron. Due to unclean equipment, analyzing of cadmium contents could not be completed.

There is convincing research which shows that the enzyme alkaline phosphatase (ALP) has a central role in the mineralization of bone, more precisely that its catalytic activity is needed in the process. ALP is found on the surface of matrix vesicles where the mineral is formed. One theory about the function of the enzyme is that it binds to fibrous collagen in the bone and thereby incorporating the mineral into the bone. The purpose of this study is to establish whether ALP binds to collagen. If this is the case, more elaborate studies around this will be performed. The strength of the binding between collagen and the different types of ALP will be evaluated, as well as on which part of the collagen the binding occurs. The binding is going to be studied by constructing a method for the ÄKTApurifier system.   Initially, the pureness of the different type of collagens was determined by using SDS-PAGE and the activity of the different types of ALP was established. These were also compared with a native PAGE. In SDS-PAGE, bovine type I collagen showed markings for a triple helix, a double helix and two single strains, α1 and α2. Bovine type II collagen showed markings for a double helix and α1-strains. Human type I collagen showed markings for a triple helix, two double helixes, two α-strains and contaminations. Trials with collagen in Native PAGE did not provide any results. However, the trials with ALP revealed that the different types of ALP had different charge.   Thereafter, blotting was performed. The results showed that all the different types of ALP, besides from E. coli, binds to bovine collagen type I and II and human collagen type I, however within various periods of time. In the trials with collagen coated plates the acquired results showed that some of the different types of ALP bind to collagen. ALP from liver binds the strongest to both collagen type I from rat and type IV from mouse. Intestinal ALP also binds to both types of collagen but not nearly as strong as liver ALP. Serum from rats did bind to collagen type I from rat but not to collagen type IV from mouse. ALP from kidney and human serum did not bind to either types of collagen. The trials concerning the ÄKTApurifier system were executed with ALP from liver alone because it had been proven to bind to bovine type I collagen through the previous methods. The results confirmed that ALP from liver binds to this type of collagen.   The conclusions from this study are that ALP does indeed bind to collagen and does so to the triple helix and double helix form as well as the single strains of collagen. In other words the part of the structure in collagen that ALP binds to must exist in all three stages of collagen formation. Furthermore, it seems like some of the different types of ALP has a higher affinity for binding to collagen, as the time for binding to collagen varies for the different types of ALP. The results differed between methods concerning different types of ALP. Although, the method we consider to give the best result was blotting. However, the method using ÄKTApurifier can be complementary but needs further development.

Today, a majority of the world’s energy need is supplied through sources that are finite and, at the current usage rates, will be consumed shortly. The high energy demand and pollution problems caused by the widespread use of fossil fuels make it increasingly necessary to develop renewable energy sources of limitless duration with smaller environmental impact than the traditional energy sources. Three fuels – rapeseed methyl ester (RME), Fischer-Tropsch (FT) diesel and FT jet fuel – derived from biomass, coal or gas were evaluated in this project. The fuel properties evaluated are in most cases listed in standards, often with recommendations, developed for biodiesel, petroleum diesel and jet fuel. Biodiesel is monoalkyl esters, e.g. RME, produced by transesterification of triglycerides in vegetable oil and an alcohol to esters and glycerin. This produce a fuel that is suitable as a direct substitution for petroleum diesel. Biodiesel may be used in pure form or in a blend with petrodiesel. Oxidative degradation and weak low temperature performance of biodiesel are properties of concern when substituting petrodiesel with biodiesel, as was shown in this project. The experiments show that oxidative stability can be improved with a synthetic antioxidant, e.g. butylated hydroxytoluene (BHT). The FT process converts syngas (a mixture of hydrogen and carbon monoxide) to a range of hydrocarbons. Syngas can be generated from a variety of carbon sources, e.g. coal, natural gas and biomass. The high-temperature (300-350 °C) FT process with iron-based catalysts is used for the production of gasoline and linear low molecular mass olefins (alkenes). The lowtemperature (200-240 °C) FT process with either iron or cobalt catalysts is used for the production of high molecular mass linear waxes. By applying various downstream processes, fuels suitable for substitution of petrodiesel and conventional jet fuel can be obtained. The FT fuels have lower densities than the conventional fuels. However, conclusions from this project are that most of the properties of FT fuels are better, or equal, than conventional petroleum fuels.

<p>Pappersbruken har två pappersmaskiner, PM4 som tillverkar mjukpapper från returfiber och PM13 som tillverkar papper till transformatorer och kablar från oblekt barrsulfatmassa. Överskottsvattnet från PM4, PM13 och returfiberanläggningen samt dagvattnet leds vidare till den externa avloppsreningen. Då någon utav pappersbruken har driftsstop erhålls ojämna utfall främst för pH och COD.</p><p>Syftet med projektet är att kartlägga pH och COD förändringen i de enskilda delstegen i processerna PM 4, PM 13 samt den externa avloppsreningen.</p><p>Det genomfördes intensiv provtagning från tolv provtagningspunkter i trettiotvå dagar. Prover analyserades samma dag med avseende på pH, konduktivitet, alkalinitet, COD, suspenderade ämnen och glödrester. Dessutom utfördes fällningsförsök på inkommande processavloppsvatten med styrd dosering av järnsulfat, järnklorid och aluminiumsulfat. Det genomfördes även metodvalidering, kalciumbestämning, samt försök med sammanblandat avloppsvattenprov.</p><p>Resultat från mätningarna visar att processlinje PM 4 har generellt sätt högre pH-, konduktivitet-, alkalinitet - och COD värden och högre halt glödrester, men lägre halt suspenderade ämnen än PM 13. Det konstaterades att den största förändringen i de två processerna sker strax innan och efter pappersmaskinen. Efter pappersmaskinen, i det vattnet som leds vidare till den externa avloppsreningen, erhålls 70% COD reduktion och en pH ökning på 0,75 pH ekvivalenter för PM 13, respektive 30% COD reduktion och en pH sänkning  på 1,27 pH ekvivalenter för PM 4. Totalt sätt, tyder resultat på att den största pH förändringen sker vid avloppsreningen, med 1,74 pH ekvivalenter. Från fällningsförsöken konstateras det att bästa pH- och lägsta COD- värden erhålls för fällningsmedlen, järnklorid och järnsulfat vid lägsta doseringsmängder, d.v.s. 97 g/m³ respektive 101 g/m³, utan pH justering. Resultat från jämförelse mellan processavloppsvatten och sammanblandat avloppsvatten, där dagvattenpåverkan utesluts tyder på 0,01-0,86 högre pH-värden i dem sammanblandade avloppsvattenprover. Alltså finns det en antydan att dagvatten bidrar till pH sänkning i ingående processavloppsvatten. För att åtgärda problemen borde järnklorid utprövas i lägre doseringsmängder i kombination med minskad svavelsyretillsats.<strong> </strong>Eventuellt kan järnklorid bytas ut mot järnsulfat. Installera mätinstrument för mätning av pH och COD direkt på det ingående- och utgående vattenflödet i den externa avloppsreningen för att möjliggöra exakt dosering av fällningskemikalierna utifrån dessa värden.</p><p>Dagvattnet borde avskiljas från avloppsreningen och istället avledas till det kommunala avloppsnätet.</p> / <p>The paper mills have two paper machines, the PM 4, producing tissue from the RCF and the PM 13, producing paper for transformers and cables from the unbleached softwood craft pulp. Excess water from PM4, PM13 and RCF factory, together with the storm water, leads on to the external wastewater treatment. When any of the paper mills has downtime, unequal outcomes are obtained, primarily with respect to pH and COD.</p><p>The aim of the project is to identify the individual steps in the processes PM 4, PM 13 and wastewater treatment re to the pH and COD changes.</p><p>Intensive sampling was carried out from twelve sampling points during thirty-two days. These samples were analyzed on the same day re to the pH, conductivity, alkalinity, COD, suspended solids, and inorganic residue. In addition, precipitation tests were carried out on the incoming wastewater with controlled doses of the ferrous sulphate, ferric chloride and aluminium sulphate. In addition methodological validation, calcium determination, and experiments with the mixed wastewater samples were carried out.</p><p>The results show that the process line PM 4 has generally higher pH, conductivity, alkalinity, COD and concentration inorganic residues, but has a lower content of suspended solids than PM 13. But the biggest difference in the two processes takes place just before and after the paper machine. After the paper machine, the water that goes on to the external wastewater treatment, where a 70% COD reduction and a pH increase of 0,75 pH equivalents for PM 13, and 30% COD reduction and a pH decrease of 1,27 pH equivalents for the PM 4 are obtained. The results suggest that the greatest pH change occurs in wastewater treatment, with 22.06%. From precipitation experiments it was found that the best pH values and lowest COD values where obtained for precipitation chemicals, ferric chloride and ferrous sulphate at the lowest dose levels, i.e. 97 g/m³ and 101 g/m³, without pH adjustment. Results from the correlation between wastewater- and mixed wastewater samples, which excludes impact of storm water, indicates that pH values are 0.01 to 0.86 pH equivalents higher in those mixed wastewater samples. Thus, there is a hint that the storm water contributes to pH reduction of the input wastewater.</p><p>To address these concerns ferric chloride should be tested at lower dose levels combined with reduced sulphuric acid addition. Eventually ferric chloride may be replaced by the iron sulphate. In addition, there should be measuring devices for pH and COD installed on the process wastewater and outgoing water, in the external wastewater treatment in order to allow for accurate dosing of chemicals precipitate from these values. Storm water should be separated from wastewater treatment and instead be diverted directly to the municipal sewage system.</p>