In the work underlying this thesis solid-phase microextraction (SPME) was evaluated as a passive sampling technique for organophosphate triesters in indoor air. These compounds are used on a large scale as flame-retarding and plastizicing additives in a variety of materials and products, and have proven to be common pollutants in indoor air. The main objective of this work was to develop an accurate method for measuring the volatile fraction. Such a method can be used in combination with active sampling to obtain information regarding the vapour/particulate distribution in different indoor environments. SPME was investigated under both equilibrium and non-equilibrium conditions and parameters associated with these different conditions were estimated. In <b>Paper I</b>, time-weighted average (TWA) SPME under dynamic conditions was investigated in order to obtain a fast air sampling method for organophosphate triesters. Among the investigated SPME coatings, the absorptive PDMS polymer had the highest affinity for the organophosphate triesters and was consequently used in all further work. Since the sampling rate is dependent on the agitation conditions, the linear airflow rates had to be carefully considered. Sampling periods as short as 1 hour were shown to be sufficient for measurements in the ng-μg m-3 range when using a PDMS 100-μm fibre and a linear flow rate above 7 cm s-1 over the fibre. SPME under equilibrium conditions is rather time-consuming, even under dynamic conditions, for slowly partitioning compounds such as organophosphate triesters. Nevertheless, this method has some significant advantages. For instance, the limit of detection is much lower compared to 1 h TWA sampling. Furthermore, the sampling time can be ignored as long as equilibrium has been attained. In <b>Paper II</b>, SPME under equilibrium conditions was investigated and evaluated for organophosphate triester vapours. Since temperature and humidity are closely associated with the distribution constant a simple study of the effect of these parameters was performed. The obtained distribution constants were used to determine the air levels in a common indoor environment. SPME and parallel active sampling on filters yielded similar results, indicating that the detected compounds were almost entirely associated with the vapour phase To apply dynamic SPME method in the field a sampler device, which enables controlled linear airflow rates to be applied, was constructed and evaluated (<b>Paper III</b>). This device was developed for application of SPME and active sampling in parallel. A GC/PICI-MS/MS method was developed and used in combination with active sampling of organophosphate triesters in indoor air (<b>Paper IV</b>). The combination of MS/MS and the soft ionization achieved with methanol as reagent gas yielded high selectivity and detection limits comparable to those provided by GC with nitrogen-phosphorus detection (NPD). The method limit of detection, when sampling 1.5 m3 of air, was in the range 0.1-1.4 ng m-3. In <b>Paper V</b>, the developed MS method was used in combination with SPME for indoor air measurements. The levels detected in the investigated indoor environments range from a few ng to μg m-3. Tris(2-chloropropyl) phosphate was detected at a concentration as high as 7 μg m-3 in a newly rebuilt lecture room.

This thesis describes the use of nuclear magnetic resonance (NMR) spectrometry as an analytical tool. The theory of NMR spectroscopy in general and quantitative NMR spectrometry (qNMR) in particular is described and the instrumental properties and parameter setups for qNMR measurements are discussed. Examples of qNMR are presented by impurity determination of pharmaceutical compounds and analysis of urine samples from rats fed with either water or a drug (metabolic profiling). The instrumental parameter setup of qNMR and traditional data pre-treatments are examined. Spectral smoothing by convolution with a triangular function, which is an unusual application in this context, was shown to be successful regarding the sensitivity and robustness of the method in paper II. In addition, papers III and IV comprise the field of peak alignment, especially designed for 1H-NMR spectra of urine samples. This is an important preprocessing tool when multivariate analysis is to be applied. A novel peak alignment method was developed and compared to the traditional bucketing approach and a conceptually different alignment method. Univariate, multivariate, linear and nonlinear data analyses were applied to qNMR data. In papers I–II, calibration models were created to examine the potential of qNMR for these applications. The data analysis in papers III–VI was mainly explorative. The potential of data fusion and data correlation was examined in order to increase the possibilities of analysing the highly complex samples from metabolic profiling (papers V–VI). Data from LC/MS analysis of the same samples were used with the 1H-NMR data in different ways. Correlation analyses between the 1H-NMR data and the drug metabolites identified from the LC/MS data were also performed. In this process, data fusion proved to be a valuable tool.

An intrinsic mathematical model for simulation of fixed bed chromatography was demonstrated and compared to more simplified models. The former model was shown to describe variations in the physical, kinetic, and operating parameters better than the latter ones. This resulted in a more reliable prediction of the chromatography process as well as a better understanding of the underlying mechanisms responsible for the separation. A procedure based on frontal liquid chromatography and a detailed mathematical model was developed to determine effective diffusion coefficients of proteins in chromatographic gels. The procedure was applied to lysozyme, bovine serum albumin, and immunoglobulin γ in Sepharose™ CL-4B. The effective diffusion coefficients were comparable to those determined by other methods. Molecularly imprinted polymers (MIPs) are traditionally prepared as irregular particles by grinding monoliths. In this thesis, a suspension polymerization providing spherical MIP beads is presented. Droplets of pre-polymerization solution were formed in mineral oil with no need of stabilizers by vigorous stirring. The droplets were transformed into solid spherical beads by free-radical polymerization. The method is fast and the performance of the beads comparable to that of irregular particles. Optimizing a MIP formulation requires a large number of experiments since the possible combinations of the components are huge. To facilitate the optimization, chemometrics was applied. The amounts of monomer, cross-linker, and porogen were chosen as the factors in the model. Multivariate data analysis indicated the influence of the factors on the binding and an optimized MIP composition was identified. The combined use of the suspension polymerization method to produce spherical beads with the application of chemometrics was shown in this thesis to drastically reduce the number of experiments and the time needed to design and optimize a new MIP.

The magnitude of an electric field possible to apply in a capillary separation system is limited, because a high electric field causes a too high current through the capillary. Application of the electric field in-line will give an increased conductivity in the column, further increasing the risk of too high currents. The conductivity changes were found to result from an overall increase in ionic strength within the electric field. The increase in ionic strength is caused by the increase in mobile phase ions with electrophoretic velocity against the flow, together with OH- or H3O+ ions (depending on polarity) formed at the inlet electrode. Further it was found that the use of a pressurized reservoir or splitting of the flow at the inlet electrode could significantly limit the conductivity changes and thereby the maximum applicable electric field strengths could be increased.

Computer simulations can be useful aids for both developing new analytical methods and enhancing the performance of existing techniques. This thesis is based on studies in which computer simulations were key elements in the development of several new tools for use in gas chromatography and dielectrophoresis. In gas chromatography, gaseous analytes are separated by exploiting differences in their partitioning between different phases, and after their partitioning parameters have been determined the separations can be computationally predicted, and optimized, for a wide range of operating conditions. Similarly, in dielectrophoresis, particles with differing polarizability or size can be separated, and since particle trajectories within a separation device can be predicted using computations, the suitability of new designs, applications of forces and combinations of operational parameters can be assessed without necessarily making or empirically testing all of the variants. Using two existing numerical methods combined with semi-empirical determinations of retention behavior, temperature-programmed gas chromatograms were predicted with less than one percent deviations from experimental data, and a new method for improving the capacity of a gas-trapping device was predicted and experimentally verified. In addition, two new concepts with potential capacity to enhance dielectrophoretic separations were developed and tested in simulations. The first provides a promising way to improve the trapping of bacteria in media with elevated conductivity by using super-positioned electric fields, and the second a way to increase selectivity in the separation of bio-particles by using multiple dielectrophoretic cycles. The studies also introduced a more accurate method for determining the conductivity of suspensions of bacteria, and a new computational method for determining the dielectrophoretic behavior of particles in concentrated suspensions. The scientific studies are summarized and discussed in the main text of this thesis, and presented in detail in seven appended papers.

Anabolic androgenic steroids are testosterone and its derivates. Testosterone is the most important naturally existing sex hormone for men and is used for its anabolic effects providing increased muscle mass. Testosterone is taken orally or by intramuscular injection in its ester form and are available illegally in different forms of esters. Anabolic androgenic steroids are today analyzed only in urine. To differentiate between the human natural testosterone and exogenous supply the quote natural testosterone and epitestosterone is used. Detection of testosterone esters in serum is an unmistakable proof of exogenous supply of testosterone. The aim of this thesis was to find a method for determining testosterone esters in serum and to study an extraction method possible for quantification of testosterone esters in serum. The technique used to separate and identify the Testosterone esters was Liquid Chromatography Tandem Mass Spectrometry Electro Spray Ionisation. Parameters for chromatography and mass detection were optimized for nine testosterone esters and evaluated according to selectivity, resolution and intensity. A method that could be used for determination of testosterone esters in serum was found. The MS-method was set and at least three possible transitions for each testosterone ester were found. The best choice of column proved to be the C18 column where all the esters were separated and seven of them were base-line separated. The C18 column along with methanol and ammonium acetate buffer, 5 mM, pH 5 showed the highest sensitivity for Multiple Reaction Monitoring-detection. A gradient profile for a total runtime of 5.6 minutes was established. Two alternative extraction procedures were tested, with tert-butylmethylether or diethyl ether/ethyl acetate and both seemed to work satisfactory. Analysis of serum proved to work well and no severe interference occurred. Results from the linearity tests indicate that future quantification method in serum will be possible.

No description available.

The objective of this master thesis was to develop an analytical method for the quantification of the cancer drug Imatinib and its main metabolite CGP74588 in plasma. Imatinib is used in the treatment of chronic myeloid leukemia and gastrointestinal stroma tumors. A quantitative analytical method was developed where reversed-phase columns with different stationary phases were studied and the sensitivity was tested with both UV detectors and a mass spectrometric detection. Since the substances were measured in plasma a solid-phase extraction was developed to purify the samples before analysis. The column chosen for the separation was the Max-RP C12 column (100 x 3 mm, 4 μm particle size) manufactured by Phenomenex with a gradient mobile phase with 1% formic acid in methanol and water. The gradient was as follows; 0 min 15:85, 7 min 60:40, 9 min 60:40 with a total runtime of 13.5 min. The internal standard chosen was Opipramol. Mass spectrometric detection using a sonic spray ionization interface in positive mode proved to be about as sensitive as UV detection at 261 nm. The generated (M+H+)+ ions were isolated and fragmented with the use of three mass spectrometric methods; one for Imatinib (transition 494 —› 394), one for CGP74588 (transition 480 —› 394) and one for Opipramol (transition 364 —› 171). For the purification of the plasma samples an Oasis HLB solid-phase extraction cartridge was selected and the recoveries were close to 100%. The developed method was partially validated and showed coefficients of variation (CV) for intra-and inter-day precision between 0.4 and 5.4% with UV detection. The validation results for the mass spectrometer were inconclusive.

A study was performed to evaluate whether established methods of analysis of protein content in barley (Kjeldahl, Dumas, or NIT (short for Near Infrared Transmittance)) gives different results for wet and dried barley. This was carried out because there are concerns regarding the well-worn NIT prediction model giving different results for these conditions and that such an error causes significant price fluctuations on the market. By performing analyses of samples, both before and after drying, of the three different barley varieties Tipple, Prestige, and Quench, with all the techniques, data was obtained that could be analyzed statistically. The study showed that the NIT prediction model gives results for wet barley that is about 0.29 percentage points higher compared to dried barley. This difference was also statistically significant when a t-test was performed. The same difference was substantially smaller for Kjeldahl and Dumas and furthermore not statistically significant. When the difference between the different barley varieties is studied a deviation of 0.34 percentage points was obtained for Tipple, 0.18 percentage points for Prestige, and 0.30 percentage points for Quench. NIT, thereby, gave results with greatest deviation when wet Tipple was analyzed. When t-tests were performed for each barley variety, it was found that results for all varieties differ significantly between wet and dry barley. A study of the difference between the crude protein content determination by Kjeldahl and Dumas was also performed by calculating the mean differences between the various methods of determination. The average difference in crude protein determination was calculated to 0.07 percentage points.

This project was aimed to study the degradation of polystyrene-butadiene-styrene block copolymer (SBS). SBS is used to modify bitumen, the binder in asphalt. From earlier studies it is known that SBS degrades in bitumen, but the degradation products have not been identified. To be able to determine the long term effects of SBS in the environment, degradation products need to be identified. Polystyrene forms rigid blocks with polybutadiene as a rubbery matrix between the ridgid blocks. When blended in bitumen, the polystyrene blocks are crosslinked to form a three dimensional network. SBS is an elastomer and has the ability to regain its shape after being a subject of mechanical force. According to the pre references the polymers had to be degraded in similar ways as they would in bitumen. Since bitumen is a very complex matrix with high boiling point and viscosity, there would be difficulties separating polymers from the bitumen. We made an assumption that it would be plausible for the degradation products formed in bitumen to form even with no bitumen present. Four different SBS polymers were used. One of the polymers was branched. Another had a high content of 1-2 polybutadiene. The third was a diblock copolymer (SB) with low styrene content compared to the others. The last was a linear SBS. The polymers were degraded in pieces of apparatus used for study ageing characteristics of bitumen as well as they were aged by refluxing in hexane. The degradation products were extracted by Solid Phase Extraction (SPE) and identified by GC-MS. Due to instrumental limits there were not many degradation products identified. The main products detected included saturated hydrocarbons in the range of 16-31 carbon atoms as well as squalene. / I det här projektet studerades nedbrytning av styren-butadien-styren block kopolymerer (SBS). SBS används för att modifiera bitumen vars användningsområde är som bindemedel i asfalt. Tidigare studier har visat att SBS bryts ned i bitumen, men nedbrytningsprodukterna har inte identfierats. För att kunna avgöra vad SBS har för långsiktiga effekter på miljön måste nedbrytnignsprodukterna identifieras. I SBS bildar polystyren styva block medan polybutadien fungerar som flexibla bryggor mellan polystyrenblocken. När SBS blandas med bitumen korslänkas polystyrenblocken så att ett tredimensionellt nätverk bildas. Eftersom SBS är en elastomer har den förmågan att återfå sin ursprungliga form efter att ha blivit utsatt för mekanisk stress. Enligt de givna förutsättningarna för studien skulle polymererna brytas ned på samma sätt som om de skulle ha varit lösta i bitumen. Eftersom bitumen är en komplex matris med hög kokpunkt och viskositet skulle det varit svårt att separera polymererna och deras nedbrytningsprodukter från bitumenet. Vi gjorde ett antagande om att det förmodligen bildas samma nedbrytningsprodukter som om bitumen varit närvarande även om nedbrytningen sker utan bitumen närvarande. Fyra olika SBS polymerer användes. En av polymererna var grenad. En annan hade en stor andel 1,2-polybutadien. Den tredje var en diblock kopolymer (SB) med lågt styren innehåll jämfört med de andra. Den sista polymeren var linjär. Polymererna åldrades i apparatur framtagen för att studera åldring av bitumen. Polymererna åldrades även genom att återloppskokas i hexan. Nedbrytningsprodukterna extraherades via fastfasextraktion (SPE) och identifierades med GC-MS. Genom instrumentella begränsningar kunde bara ett fåtal nedbrytningsprodukter identifieras. Bland de produkter som identifierades fanns alkaner och alkener med mellan 16 och 31 kolatomer, men även skvalen identifierades som nedbrytningsprodukt. Lågmolekylära ämnen kan ha ventilerats bort för att vi inte hade tillgång till apparatur för adsorbtion av flyktiga ämnen för injektion i GC-MS.