Whey protein isolate (WPI) contains at least 90% protein and should ideally possess a bland flavor without typical dairy flavors including sweet aromatic and cooked/milky notes. However, its flavor may be highly variable due to factors including original whey source, processing and storage conditions. Novel technologies removing nonpolar compounds responsible for off-flavors and off-flavor formations are desirable. The major objective of this research was to evaluate impacts of supercritical carbon dioxide (scCO2) extraction, a known green process, on volatile profiles of WPI. A prior sub-objective was to establish an analytical technique for characterization of volatiles. Specifically, adsorption conditions in a well-established head-space solid-phase microextraction (SPME) method were used for quick and reliable assays of volatiles in WPI, using a divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS) fiber. The adsorption of volatiles on the SPME fiber was studied at 21, 40 or 50 °C, each with durations of 5, 15 and 20 min, and analyzed by GC/MS. Based on the number of GC/MS peaks and the corresponding peak areas, adsorption conditions of 50 °C for 20 min were selected for subsequent studies. In the second sub-objective, GC/MS profiles of WPI were characterized after scCO2 extraction using a continuous stream of CO2 at 50 g/min, controlled at various combinations of temperature (30-65°C), pressure (7.0-30.0 MPa), and duration (10-90 min). Extractions with a higher temperature and a higher pressure for a longer time were generally more effective in removing volatiles, and most peaks on the chromatogram of the unprocessed WPI sample disappeared or were reduced very significantly after all studied extraction conditions, even at subcritical conditions of 7.0 MPa and 30 °C for 1 hour. Our findings demonstrated that supercritical or subcritical CO2 may provide a green approach to reduce volatiles in whey protein preparations for novel food applications.

Aldehydes in lung cancer cell culture have been investigated using gas chromatography/mass spectrometry and solid-phase microextraction with on-fiber derivatization. In this study, the poly(dimethylsiloxane/divinylbenzene (PDMS/DVB) fiber was used and o-2,3,4,5,6-(pentafluorobenzyl) hydroxylamine hydrochloride (PFBHA) was first loaded on the fiber. Aldehydes in the headspace of lung cancer cell culture were extracted by solid-phase microextraction (SPME) fiber and subsequently derivatized by PFBHA on the fiber. Finally, the aldehyde oximes formed on the fiber were analyzed by gas chromatography/mass spectrometry (GC/MS). Using this method, acetaldehyde decrease was found in both non-small lung cancer cell cultures studied compared to the medium control study. The results of spiking the cell culture with acetaldehyde solution showed that 5 million SK-MES-1 cell lines could consume up to 4.5 uM acetaldehyde in 15-ml medium, and 5 million NCI-H522 cell lines could consume 5.9 uM acetaldehyde in 15-ml medium. The decrease of acetaldehyde may contribute to the metabolism of lung cancer cells. It was proved that GC/MS and SPME with on-fiber derivatization is a simple, rapid, sensitive and solvent-free method for the detection of aldehydes in lung cancer cell culture.

The development of a synthetic method of radiolabelled methylphosphono-fluoridates on a milligram scale is presented. The aim of this method is, besides affording high yield, to choose reaction pathways and reagents so that handling and transfer of labelled toxic substances is minimised, thereby reducing the risk of exposure as much as possible. The only substituent that is stable enough to be labelled is the methyl group, directly bonded to phosphorus. A drawback when labelling the methyl group is that it requires the label to be introduced early in the synthesis since the carbon-phosphorus bond of the methyl substituent usually has to be synthesized a few steps ahead of the final product. Two new classes of reagents for halogenation of phosphorus oxyacids have been developed. Firstly, four different analogues of α-chloroenamines and α-fluoroenamines were evaluated. Secondly, cyanuric fluoride was assessed in solution, but more importantly, as a resin-bound reagent. The reagents are evaluated for halogenation of phosphinic, phosphonic and phosphoric acids. Cyanuric fluoride is also successfully loaded on a polystyrene resin and used as a solid-phase reagent. The reagents produce high yields and low levels of impurities on a milligram scale. Furthermore, a new method for the preparation of mono-alkylated phosphonic acids on a small scale has been developed. The new method utilises the crystal water bound to certain salts to liberate limited amounts of water in a controlled manner. Phosphonic dichlorides are in this way reacted with water to form anhydrides. The anhydride is then cleaved with an appropriate alcohol to produce mono-alkylated phosphonic acids. / Rikard Norlin = Rikard Wärme

This thesis investigates three different surface modifications, and the route to design and synthesize them. The thesis is therefore divided into three sub- projects. (i.) Design and synthesis of a peptide which secondary structure could be controlled by a negatively charged surface. (ii.) Design and synthesis of a cyclic peptide, that would self-organize prior to surface interaction, using the type I anti-freeze protein of a winter flounder as template. (iii.) The use of solid-phase synthesis to make the synthesis of SAM-molecules easier.

Nitrogen is known to be a limiting factor in polluted environments, however many studies overlook the potential role for nitrogen to significantly influence the removal rate and efficiency with which microorganisms can degrade aromatic hydrocarbons. In this study, inoculated and uninoculated aerated soil microcosms containing different s-triazines were examined for their ability to efficiently and rapidly treat contaminated soils containing naphthalene, nitrobenzene, and toluene (NNT), via a microbial consortium consisting of Pseudomonas, Rhodococcus, and Aeromonas. After an experimental period of 14 days, greater than 90% degradation of NNT supplemented with different s-triazines, at concentrations of 1000-3000 ppm was observed. A difference in the degradation of NNT was seen in inoculated box reactors supplemented with cyanuric acid, melamine, and atrazine in comparison to uninoculated box reactors. Combined usage of 16s rDNA and 16s rRNA analysis was then applied to study the bacterial communities, and determine the abundance and survival of inoculated strains within box reactors contaminated with NNT. The bacterial diversity within clone libraries obtained illustrated a dominance of proteobacteria and gram positive bacteria. Analysis from clone libraries also showed that inoculated strains did survive within each condition, but were not the most predominant members present in the communities. This research shows that significant removal of NNT can be achieved in two weeks with the supplementation of one of the s-triazines. However, differences in degradation and the microbial populations present within contaminated communities will be seen depending on which nitrogen sources are used and whether or not environments are bioaugmented or not.

Various needle trap devices (NTDs) with different designs for different applications have been developed during the past decade. A theoretical model on the fundamentals of the NTD was recently proposed, which employed the theory of frontal (gas-solid) chromatography to describe the sampling process, where a gaseous sample was continuously introduced into the sorbent bed. In this investigation, different types of sorbent particles with different dimensions were packed into the needle as adsorbents. The effect of particle dimension, which would affect the packing density and consequently the capacity, the extraction efficiency, and desorption efficiency of the NTD were experimentally investigated and the proposed theory was validated. The results demonstrated that NTDs packed with small particles possess higher extraction capacity and efficiency but much higher resistance to flow as well. The higher resistance did not necessarily result in poor desorption efficiency, because desorption efficiency was affected by both the sorbent bed structure and the desorption gas flow. The relationships observed among those physical parameters provide valuable guidance on how to design an NTD with high performance potential for future applications. For particulate sampling, it was found that NTDs packed with different particles presented high collection efficiency of the particulates being investigated, and the collection efficiency was dominated by the pore size and distribution of the sorbent bed packed inside the needle. Collection efficiency also increased with increase in solidity of the sorbent bed; the increase in humidity of the aerosol sample; and the decrease of sampling rate. The results also provide valuable guidance on the optimisation of needle trap for particulate collection.

Biocides are widely used as a preservative or as an antiseptic agent in consumer care products such as toothpaste, mouthwash, and soaps, as well as in household cleaners and even in textiles due to their high antimicrobial effectiveness. The usage of this compounds results in discharge to wastewater treatment plants and so into surface waters. Their existence in the environment is of importance due to their negative effects on aquatic environment microorganisms and human health in terms of occurrence in surface waters and their fate in wastewater treatment plants. In this scope, this study focuses on occurrence and fate of selected biocides, namely triclosan (TCS) and chlorhexidine (CHD), in wastewater treatment plants and in surface waters. It was aimed to determine the biocides levels in surface water and wastewater in Turkey. For the wastewater treatment plant (WWTP) studies, several WWTPs with different process configurations, namely, Tatlar WWTP, METU WWTP, Kayseri WWTP and Antalya WWTPs were selected. Composite wastewater samples were taken from various points along the WWTPs on a seasonally basis for one year period. For the surface water part, samples were taken monthly from three different sources with different pollution levels, namely, Kesikk&ouml / pr&uuml / Reservoir, &Ccedil / amlidere Reservoir and Eymir Lake for one year period. All water samples were analyzed for their biocide level using liquid chromatography, following solid phase cartridge extraction. As a result of analyses, TCS concentration in surface water samples was detected as in the range of 0.65-11.15 ng/L, 0.86-48.96 ng/L and 0.86-757.7 ng/L for clean, moderately polluted and polluted water sources respectively. The recovery of solid phase extraction analyses for TCS was achieved as %92. CHD concentration was determined as in the range of &lt / 1.33-5.31 ng/L for surface water samples and the recovery of extraction were calculated as %96 for CHD. The concentration of TCS in wastewater samples was measured as in the range of 1.77-94.47 ng/L and 1.40-15.09 ng/L for influent and effluent samples respectively. These ranges became 1.39-10.45 ng/L and &lt / 1.32-2.44 ng/L for CHD. The highest concentrations of biocides were observed in sludge samples with concentrations of 1117-3687 &mu / g/kg and 510-2742 &mu / g/kg for TCS and CHD. Biocide removal efficiency of primary and biological treatment together was reported as % 67.5&plusmn / 8.2 in January 2012 Tatlar WWTP analyses.

La tesis doctoral está dividida en 3 partes:La primera presenta la aplicación del grupo TCP a la síntesis de PNAs en fase sólida. La segunda está dedicada a la síntesis de un nuevo tipo de nucleoaminácido para la síntesis de nuevos nucleopéptidos. La tercera está centrada en los estudios iniciales de síntesis de derivados de Pantocina B, un compuesto con propiedades antimicrobianas. / The doctoral thesis is divided in 3 parts: The first presents the application of the TCP group to the synthesis of PNAs in solid phase. The second is dedicated to the synthesis of a new type of nucleoaminácid for the synthesis of new nucleopéptids. The third is centered in the initial studies of synthesis of derivatese of Pantocina B, a componend with antimicrobial properties.

A cold fiber solid phase microextraction device was designed and constructed based on the use of a thermoelectric cooler (TEC). A three-stage thermoelectric cooler was used for cooling a copper rod coated with a polydimethylsiloxane (PDMS) hollow fiber, which served as the SPME fiber. The copper rod was mounted on a commercial SPME plunger and exposed to the cold surface of the TEC, which was enclosed in a small aluminum box. A heat sink and a fan dissipated the generated heat at the hot side of the TEC. By applying an appropriate DC voltage to the TEC, the upper part of the copper rod, which was in contact to the cold side of the TEC, was cooled and the hollow fiber reached a lower temperature through heat transfer. A thermocouple was embedded in the cold side of the TEC for indirect measurement of the fiber temperature. A portable cold fiber SPME device was made by using a car battery as the power supply. The cold fiber SPME device with thermoelectric cooling was applied in quantitative analysis of off-flavors in rice. Hexanal, nonanal, and undecanal were chosen as three test analytes in rice. These analytes were identified according to their retention times and analyzed with a GC/FID instrument. Headspace extraction conditions (i.e. extraction temperature and extraction time) were optimized. Standard addition calibration graphs were obtained at the optimized conditions and the concentrations of the three analytes were calculated. The developed method was compared to a conventional solvent extraction method. The applicability of the portable cold fiber SPME with TEC for field sampling was tested. The effect of cooling on extraction recovery and the reproducibility of extraction were examined for extractions from an n-alkane flow through system. It was found that the extraction recoveries were significantly higher when the fiber was cooled. To further investigate the effect of cooling on the sensitivity of SPME in field sampling, the portable cold fiber SPME was used for extraction of volatile components from living wisteria flowers. Both the number of identified compounds and the related peak areas increased for extractions with cold PDMS fiber relative to without cooling and commercial PDMS and PA fibers. The portable cold fiber SPME device was also used for field sampling of volatile components of living lily-of-the-valley flowers and the extracted compounds were analyzed with GC/MS. The desorption kinetics of hydrophobic organic compounds (HOCs) from environmental solid matrices was investigated using cold fiber SPME with CO2 cooling. Polycyclic aromatic hydrocarbons (PAHs) and selected volatile organic compounds (i.e. toluene, ethylbenzene, o-xylene) were used as test analytes. Sand, silica gel, and clay were used as laboratory model solid matrices and were contaminated by the test analytes. Certified sediments were used as naturally contaminated samples. In this approach, the organic compounds, released from contaminated solid samples at different elevated temperatures, were exhaustively extracted with cold fiber SPME over different extraction times. The extraction data were used to obtain desorption and Arrhenius plots. The rate constants of desorption and activation energies of desorption were measured for each contaminant using these plots. The results were comparable to those reported in the literature.

The aim of this study was to develop methods for sample preparation for 8-OHdG in blood plasma samples with different solid phase extraction techniques using HPLC with an elec- trochemical detector. The solid phase extraction cartridges used were Chromabond® C18, Oasis® MAX, and three types of SupelMIPTM cartridges for chloramphenicol, riboflavin, and nitroimidazoles. The SupelMIPTM cartridges are based on molecularly imprinted polymers- technique. The separation of 8-OHdG in samples extracted from blood plasma was carried out with a Thermo Quest Hypersil Division ODS column (250 mm × 4 mm, 3μm I.D.) and methanol:buffer (10:90, v/v) as mobile phase. Recovery and selectivity was studied for the different solid phase extraction methods. The highest recovery was obtained using the Chromabond C18 cartridge with a recovery of 92%, and CV coefficient 9.5% (n = 4). 8-OHdG could not be extracted on MIP-cartridges for chloramphenicol or riboflavin, but was retained on MIP columns for nitroimidazoles, and the highest recovery was 49%.