Analytical instrumentation for quantitative in vivo stable isotope metabolic studies has included gas chromatography-mass spectrometry (GC-MS) and gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). Limitations of existing MS techniques include excessive parent ion fragmentation, time-consuming sample preparation, and complex instrument operating parameters. In this thesis, procedures for set up and implementation of four GC-MS techniques are described. The first three replicate existing GC-MS single quadrupole methods on an ion trap, and are electron ionization (EI), external methane positive chemical ionization (PCI), and methane negative chemical ionization (NCI). The fourth method is a novel GC-MS mass-selected ion trap internal isobutane positive chemical ionization technique. Four groups of rats were administered isotopic linoleic acid, and liver tissue was collected for labelled linoleic and n-6 polyunsaturated fatty acids (PUFA) metabolites analyses. Qualitative utility of EI was confirmed, and its quantitative limitations exposed. Labelled 18:2n-6 and n-6 PUFA metabolites were detected using external methane PCI, though limited due to significant fragmentation. Methane NCI also detected labelled 18:2n-6, as well as minimizing fragmentation. However, time-consuming sample preparation and non-linear responses were major limitations. Internal isobutane PCI was useful in detecting isotopic 18:2n-6 and n-6 PUFA metabolites. Fragmentation was reduced compared to EI and external methane PCI. Limitations include fragmentation of HUFAs such as EPA and DHA. The novel internal isobutane PCI is more sensitive than external methane PCI and NCI, produces highly linear responses, is simpler and less expensive to operate than C-IRMS, offers reliable instrument operation, and sample preparation time is minimal. Regular quantitative analyses of HUFAs such as EPA and DHA may require further refinements such as using lower energy reagents than isobutane, including acetonitrile and ammonia.

Needle trap device (NTD) is a technique that is useful for a wide variety of applications involving the sample preparation of compounds with a wide range of chemico-physico properties, and varying volatilities. A newly designed NTD that improves the performance relative to previous NTD designs is simple to produce is developed. The NTD utilizes a side-hole needle with a modified tip to improve the sealing between the NTD and narrow neck liner of the GC injector, thereby increasing the desorption efficiency. The slurry packing method was applied, evaluated, and NTDs prepared by this method were compared to NTDs prepared using the vacuum aspiration method. NTD geometries including blunt tip with a side-hole needle, tapered tip with side-hole needle, dome tapered tip with side-hole, sliding tip with side-hole and blunt tip with no side-hole needle (expanded desorptive flow) were prepared and evaluated. Sampling performance and desorption efficiency were investigated using automated headspace extraction of benzene, toluene, ethylbenzene, p¬-xylene (BTEX), anthracene and pyrene. The tapered tip and sliding tip NTDs were found to have increased desorption efficiency. SPME and NTDs are valuable sample preparation tools for on-site analysis. Combining both extraction techniques allows for the differentiation of free and particle-bound compounds in a sample matrix. Portable GC/MS instrumentation can achieve fast separation, identification, and quantitation of samples prepared by the above techniques on-site without the need for transport to the laboratory. This minimizes the effects of volatiles lost and sample degradation during storage time. Here, SPME and tapered tip NTDs combined with portable GC/MS are used to investigate free and total emissions of BTEX and select PAHs from gasoline and diesel exhaust. Using the above optimized technologies, cigarette smoke in a smoking area where people were actively smoking and inside a smoker’s car were also investigated. Target contaminants were found in the investigated matrices at ng/mL levels.

New energy resources that can replace petroleum diesel are needed and biodiesel may be an excellent substitute. During storage and use, biodiesel can oxidize and form peroxides, leading to degradation of the fuel. It is therefore important to determine the peroxide content of the fuel. In this thesis the triphenylphosphine method, which involves GC-MS analysis has been compared to the standard iodometric method which involves titration with sodium thiosulfate. Working standards were made to see how well the iodometric method worked and also in order to make a calibration curve in GC-MS. Aged samples were then analyzed. The results showed that both methods gave good linearity and repeatability with limits of detection (LOD) and quantification (LOQ) of 0,06 mM and 0 17-0,65 mM respectively. However, the iodometric method seemed more circumstantial since it needed a lot of solvents and different reagents. The triphenylphosphine method was on contrary very simple to use and did not require too much preparation work. The latter also showed better reproducibility. In conclusions, both methods can be used in evaluating the peroxide content of biodiesel, but the triphenylphosphine method has the advantages of higher simplicity and better reproducibility.

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第23回名古屋大学年代測定総合研究センターシンポジウム平成22(2010)年度報告

Chlorine gas bleaching was a common method used in pulp industries. As a consequence, significant amounts of chlorine were discharged into surrounding aquatic ecosystems, affecting the biota. Chlorinated organic pollutants are formed when chlorine react with organic material. Octadecanoic acid (stearic acid) is one of the most common saturated fatty acids in aquatic biota. In a naturally occurring process two and four chlorine atoms, respectively, are added over the unsaturated bonds, forming 9,10-dichloro octadecanoic acid and 9,10,12,13-tetrachloro octadecanoic acid. These are the chlorinated fatty acids (ClFA) under investigation in this Bachelor’s Thesis. The methodological framework for measuring ClFA is investigated in this essay. The scope is to evaluate the method of isolating and quantifying the compounds as described in Åkesson-Nilsson’s (2004) dissertation. The method includes: extraction of the lipid, transesterification (where the fatty acids, including the ClFAs, are separated from the lipids and transformed into their respective methyl esters through two methods, acidic catalysis with BF3 or H2SO4), separation (by solid phase extraction) and determination of ClFA concentration with a halogen specific detector (GC-XSD/MS). Furthermore, the scope is to investigate collected fish samples (from Norrsundet) with the abovementioned method. By making a dilution series with known concentrations it was possible to establish calibration curves, to give in an indication of the effectiveness of the method. BF3 is in need of updating due to being experienced as slower and less stable than the H2SO4-method. However, it was concluded that the H2SO4-method was more effective on the standard samples and that the BF3-method was more effective on the fish lipid samples. In one of the lipid samples (lavaret transesterified with BF3) a detectable concentration of 9,10,12,13-tetrachloro octadecanoic acid was discovered. Therefore, we question SEPAs decision to cancel investigations in Norrsundet. Our results could indicate that ClFAs are still an issue that could affect the ecosystem’s biota.

En metod som kan användas för att extrahera opolära narkotika- och läkemedelssubstanser ur matriser med ett högt fettinnehåll har tagits fram. Metoden är baserad på ett extraktionsprotokoll från Livsmedelsverket (SLV K1-f4-m018.3) som används för analys av bekämpningsmedel i animaliska livsmedel. Metoden har sedan optimerats för forensiska applikationer genom att ändra olika variabler. Två av variablerna som testades var olika typer av extraktionsmedel och mängden tillsatt C18 (oktadekylsilyl)-sorbent. Experimenten visade att acetonitril som extraktionsmedel och en ökad tillsats av C18 eliminerade den största andelen av fettmolekylerna i matrisen utan att kompromissa med utbytet av den undersökta analyten. Analyterna som undersöktes var 5F-AKB48, tetrahydrocannabinol (THC) och alprazolam och den fettinnehållande matrisen utgjordes av smör. Metoden lyckades minska fetthalten i smörproverna från ca 80- till 3-5 % fett samtidigt som ett utbyte av analyterna erhölls på 70-90 %. Analyten med högst utbyte efter extraktionen var 5F- AKB48 med ett utbyte på cirka 90 %. Utbytet för alprazolam och THC blev 72- respektive 75 %. Vid extraktion med etylacetat, som var det extraktionsmedel som användes i den ursprungliga metoden (SLV K1-f4-m018.3), erhölls inga minskningar av fetthalten överhuvudtaget utan hamnade runt 80 % i samtliga analyser. När metodens kapacitet testades med ökade fetthalter visade det sig att fetthalten efter extraktionen blev lägre i de prover som hade en högre fetthalt från början. Utbytet av analyten (5F-AKB48 i detta fall) påverkades däremot inte nämnvärt av de ökade fetthalterna utan höll sig stabilt runt 85 %.

Long-term dynamics of lake-water total organic carbon (LW-TOC) concentrations in freshwater lakes provide an important perspective on the recent increases in LW-TOC observed in many of these systems and may assist with the identification of natural and anthropogenic drivers of change. This study examines how LW-TOC in Dragsjön, a lake situated in an area with a long history of anthropogenic land use, has changed in response to natural and anthropogenic perturbations throughout the Holocene. To provide a better understanding of the processes involved, a multi-proxy study was conducted and included multi-element geochemistry (17 major and trace elements), biogenic silica, organic matter (OM) content and composition, and the faecal biomarker “coprostanol”. The direct biomarker for anthropogenic presence, “coprostanol”, and a detailed characterisation of OM composition are for the first time applied for tracing changes in LW-TOC. Natural processes contributed to stable LW-TOC concentrations in Dragsjön for most part of the Holocene. Humans were present in the catchment from AD 100 as indicated by coprostanol, but did not begin to affect LW-TOC until c. AD 1500. In the last 500 years LW-TOC steadily declined from 17 to 10 mg L-1 in response to anthropogenic alterations to the terrestrial biomass balance. The increase in LW-TOC during the last 70 years likely represents a recovery from anthropogenic disturbance rather than a baseline shift in response to any of the number of proposed recent stressors. The faecal biomarker coprostanol and OM composition provided information essential for identifying and characterising the effects of anthropogenic disturbance.

Plastic debris are accumulating in our oceans and are degraded into smaller pieces which eventually becomes small enough to be available to lower thropic level organisms. Microplastics, commonly defined as plastic particles <5 mm, are globally distributed and found at remote locations far away from industrialized and populated areas. The effects of macro sized plastics is well understood whilst the effects of microplastics is hard to predict. It is known that microplastics act as transfer vectors for a wide range of toxic chemicals into organisms, and it is also known that the particle itself can cause toxic responses such as increased immune response and endocrine disruption. Researchers utilize virgin plastic pellets in order to determine the toxicological effect of the plastic particle itself, but resent research suggest that these virgin plastics may release chemicals that contribute to the toxic response and thus complicates the interpretation of the results. In present study, five different virgin plastics were allowed to leach in artificial seawater under conditions that mimic those used in particle toxicity studies. Plastics included were polyethylene, polypropylene, polyvinyl chloride, high-density polyethylene and polystyrene. Leachable monomers and oligomers were found in three of the five plastics tested: polyvinyl chloride, high-density polyethylene and polystyrene. Leached compounds from polyvinyl chloride were not identified due to time limitations. Aliphatic hydrocarbons in the size C14-C22 were leached out from high-density polyethylene in the concentration range 0.47 × 10-3 – 1.13 × 10-3 μg ml-1 within 24 hours. Polystyrene was found to leach styrene monomer which reached a concentration of 0.17 μg ml-1 within 24 hours.

Five sediment samples were collected in Bua on the Swedish west coast, near two industries, a paper mill, and a nuclear power plant. The two industries use water in their processes and have long been associated with releases of different substances, such as PCBs, and other chlorinated compounds. The environmental impact by the two industries is believed to be significant. The aim of the project was to examine the sediments close to both the water intake and water output to determine if these industrial activities have in any way changed the composition of the sediments. The sediments were extracted by Soxhlet extraction, followed by a deactivated silica and an acidic silica clean-up and then analysed by using a gas chromatograph coupled to a mass spectrometer, (GC-MS) with electron ionization, EI+, mode used in full scan mode. Each mass spectra were analysed by comparing them to the NIST database from 1998. The results were inconclusive since the peaks were not properly resolved, causing a poor correlation to the NIST database. One batch was specifically analysed for polychlorinated biphenyls (PCB) by using an atmospheric pressure gas chromatograph (APGC) coupled to a mass spectrometer (MS). The PCB analysis provided accurate results, except for the Ringhals intake where the MS became saturated due to the high levels. The river Viskan also showed high levels of PCB. The congener pattern from PCBs found near Ringhals intake resembled an Aroclor pattern from Aroclor 1248. Since the Aroclor pattern is only seen in Ringhals intake, the source is most likely from the small harbour and not from either of the industries.