No description available.

No description available.

The National Air Emission Monitoring Study (NAEMS) project was funded by the Agricultural Air Research Council (AARC) to evaluate agricultural emissions nationwide. Utah State University (USU) is conducting a parallel study on agricultural emissions at a Cache Valley poultry facility. As part of this parallel study, samples of animal feed, eggs and animal waste were collected weekly from three manure barns (designated: manure barn, barn 4 - manure belt and barn 5 - high rise) from May 2008 to November 2009. These samples were analyzed to determine ammonia content, total Kjeldahl nitrogen content and ammonia emission. The yearly average calculated NH3 values for manure barn, barn 4 and barn 5 were determined in units of mg NH3/gmanure as: 1.1 ± 0.2, 0.6 ± 0.1 and 0.8 ± 0.1, respectively. The yearly average calculated TKN values in units of % N were determined as: 2.0% ± 0.3, 1.6% ± 0.3 and 1.9% ± 0.3 for manure barn, barn 4 and barn 5, respectively. The yearly average of NH3 emission in units of mg NH3/bird-day was determined to be 440 ± 180 mg NH3/bird-day for barn 4, and 540 ± 190 mg NH3/bird-day for barn 5. The ammonia and organic amines emissions in ambient air at a Cache valley confined poultry facility were measured by using a sulfuric acid trapping solution in an impinger train followed ion chromatography (IC) detection. The yearly average concentrations of ammonia in ambient air at the barns were calculated at 11.9 ± 2.9 ppm at the manure belt barn and 12.7 ± 3.1 ppm at the high rise barn. No organic amines were detected in the collected ambient air samples by the ion chromatography method. Because there were no amines detected by the IC method, limits of detection of organic amines in air were studied. The results showed that the organic amines in the manure must occur at a minimum concentration of 1 ppm in order to have sufficient vapor pressure so that enough is transported to the impingers for trapping and subsequently be detected by the IC.

No description available.

No description available.

<p>Cyklosporin A (CsA) är en cyklisk polypeptid med molekylvikten är 1202.6 Da. Substansen har svampursprung (<em>Tolypocladium inflatum Gams)</em> och starka immunhämmande egenskaper. CsA används därför som immunsuppressivt läkemedel för att förhindra avstötning av transplanterade organ och benmärg, samt vid behandling av graft-versus-host-disease (transplantat-mot-värd-sjukdom). CsA har ett snävt terapeutiskt fönster, vilket betyder att skillnaden mellan effektivitet och toxicitet är liten. Biverkningarna av substansen är många och en del av dem allvarliga, såsom nedsatt njurfunktion och ökad risk för utvecklande av diabetes och maligna sjukdomar som exempelvis lymfom. Den inter- och intraindividuella variabiliteten i farmakokinetik och farmakodynamik är dessutom stor. Det är därför ytterst viktigt att följa behandlingen med koncentrationsbestämningar av CsA i helblod.</p><p>Det finns ett flertal olika analysmetoder för CsA tillgängliga, såsom immunoassays, vätskekromatografi (HPLC) och vätskekromatografi-tandem-massspektrometri (LC-MS/MS). Avdelningen för klinisk kemi vid Centralsjukhuset i Karlstad har sedan många år använt en radioimmunoassay, CYCLO-Trac SP^®, från DiaSorin för att bestämma CsA i helblod. Laboratoriets önskan är att ersätta denna metod, vilken använder radioaktiva isotoper, med en snabbare och mer selektiv LC-MS/MS-metod. </p><p>I detta arbete har en LC-MS/MS-metod för analys av cyklosporin A i helblod utvecklats och validerats. Metoden har snabb provupparbetning och kromatografi och använder positiv elektrospray som joniseringsteknik. Två procedurer för proteinfällning utvärderades som provupparbetningsförfarande under metodutvecklingen och två olika internstandarder testades; CsA analogen cyklosporin D och isotopmärkt CsA (d₁₂-CsA). </p><p>Efter den fullständiga valideringen infördes metoden i rutinarbetet 2009-11-01. Resultat från både LC-MS/MS och den radioimmunologiska metoden lämnas ut parallellt under minst fem månader.</p> / <p>Cyclosporine A (CsA) is a cyclic undecapolypeptide of fungal origin (<em>Tolypocladium inflatum Gams</em>). It has a molecular weight of 1202.6 Da and is used as an immunouppressive drug to prevent rejection of transplanted organs and bone marrow, and for the treatment of graft-versus-host disease. CsA exhibits a narrow therapeutic range between efficacy and toxicity. There are many side effects exerted by the drug and some of them are serious, such as renal dysfunction and increased risk of developing diabetes and malignant diseases such as lymphoma. In addition, the inter-individual and intra-individual pharmacokinetic and pharmacodynamic variability is large. Constant monitoring of the CsA-concentration is therefore mandatory. </p><p>There are several analytical methods available for the determination of CsA, such as immunoassays, liquid chromatography (HPLC) and tandem mass spectrometry (LC-MS/MS). The department of Clinical Chemistry at the Central Hospital in Karlstad has for many years used a radioimmunoassay, the CYCLO-Trac SP^® from DiaSorin, for CsA-determinations. The laboratory wants to replace this method, which uses radioactive isotopes, with a faster and more selective LC-MS/MS method. </p><p>In this work a LC-MS/MS method, utilizing positive electrospray, with a fast sample preparation and chromatography for the determination of CsA in whole blood has been developed and validated. Two protein precipitation procedures were evaluated for sample preparation during the method development and two different internal standards were tested; the CsA analog cyclosporine D (CsD) and an isotope labelled CsA (d₁₂-CsA). The LC-MS/MS assay was fully validated and implemented in the routine work at the laboratory on November 1 2009. Results from both the CYCLO-Trac SP^® method and the LC-MS/MS assay will be reported for at least five months.</p>

Spectroscopic imaging is a vital tool for studying heterogeneous samples such as bacteria and tissue. Its ability to acquire spatially resolved information allows for identification and classification of the various constituents within a sample. Spectroscopic imagers quickly acquire thousands to tens of thousands of spectra per measurement. These data are often arranged in the form of a 3-dimensional (3D) data cube which contains two spatial dimensions and one spectral dimension. This large amount of data is beneficial for gaining a thorough understanding about the distributions of chemical information. If too little information is measured, important chemical behavior may be overlooked. Statistical analysis algorithms (chemometrics) are required to determine the relevant spectroscopic information within a data cube. Applying chemometrics to such large volumes of data presents computational difficulties regarding computer memory and processing speed. To overcome these burdens, wavelet transform compression is applied prior to chemometric evaluation to accelerate computations and reduce data storage requirements. To optimize compression by enhancing acceleration and reducing approximation errors, different wavelets, or „hybrid wavelets‟, can be applied to the different dimensions of a 3D data set. Determining which combination of wavelets will yield the most compression and best data representation is difficult since many possibilities exist. A compression method is presented that automatically determines the optimum wavelet combinations for a given data set. Principal component analysis (PCA) is used to demonstrate the capabilities of this new procedure, but the compression routine is advantageous for many chemometric techniques. Although linear algorithms like PCA work well in many situations, they are not well-adapted for explaining nonlinear relationships. Kernel principal component analysis (KPCA) has recently been developed to overcome the limitations of linear algorithms. However, when applied to spectroscopic imaging, KPCA calculations require multiple gigabytes of RAM just for holding the data. Therefore, routine use of the algorithm is often prohibited on personal computers. To circumvent such situations, a wavelet compression algorithm is presented that avoids ever having to hold all data in memory at any point during the calculations. The goal is to enable the application of KPCA to large imaging data sets of heterogeneous samples.

Raman spectroscopy is normally a non-destructive, highly selective technique that has become an ubiquitous tool for analytical chemists. One of the primary limitations of Raman spectroscopy, however, is the relatively low cross-section of the technique. With signal enhancements relative to normal Raman scattering as high as 1011, the ultra-trace detection of adsorbates down to the single molecule level has been achieved with SERS. Despite the dramatic improvement in the sensitivity and the high selectivity afforded by the SERS method, the acceptance of SERS as a general analytical tool has been hindered by a lack of stability and reproducibility in the substrates. This lack of stability has been particularly troublesome because unstable substrates exhibit reduced shelf lives as well as a reduced ability to monitor processes that occur under non-ideal conditions such as high temperature or harsh chemical environments. In this thesis, two different works are reported that address the two major hurdles facing the SERS field in the development of a stable and reproducible SERS substrate. First, the development of a SERS-active substrate that exhibits improved temporal and thermal stability and is capable of in-situ high temperature measurement of analytes adsorbed on the surface is presented. The substrates are prepared by depositing an ultra-thin layer of alumina by Atomic Layer Deposition (ALD) onto silver island films grown by thermal evaporation. We demonstrate the application of alumina-coated substrates to the measurement of the dehydration of trace amounts of calcium nitrate tetrahydrate as a function of temperature. As a development of the above mentioned work, the combination of a silver/gold layered architecture obtained by thermal evaporation with an ultra-thin alumina overlayer to generate a re-usable SERS substrate that is simple, relatively inexpensive and stable is reported. The relative thicknesses of the silver and gold and the alumina overlayer was optimized to deliver the maximum SERS enhancement and optimal stability when the substrate was subjected to high temperature. Utilizing the method of thermal desorption of the analyte, the substrate surface is regenerated and able to be reused multiple times with little reduction in SERS activity. Second, in the development of a reproducible SERS substrate, the application of monodisperse silver nanocube colloidal substrate in microfluidic SERS is demonstrated. In static SERS experiments, one often has to search for “hot spots”, which are positions of a drastically increased SERS signal compared to the rest of the probe volume, in an inhomogeneous solution. To overcome this problem and prevent the decomposition and or fragmentation of SERS substrate and analyte, respectively, the implementation of flow cell is a promising way. At the beginning analyte, colloidal solution and aggregation agent were brought into a mixing chamber, where they were thoroughly mixed before being directed to a sample cell for detection. With this method, a relatively high amount of sample volume is necessary. In addition to the advantages listed above, efforts have been made to reduce the required amount of the sample solution by the design of low-cost poly (dimethylsiloxane) chips via soft lithography technique. The sample solution is passively pumped through the microfluidic channel, where an optical detection window is implemented for acquisition of a SERS spectrum.

This thesis focuses on the use of molecularly imprinted polymers as selective sorbents for solid-phase extraction (MISPE). The MISPE methods developed were mainly intended for use with biological samples, such as human urine and blood plasma. These body fluids are complex samples, which often need an effective clean-up step before analysis to reduce the levels of possible interfering substances from the matrix, especially if the analytes are present in trace amounts. Solid-phase extraction (SPE) is a well-established and routinely used method for clean-up and preconcentration of samples from diverse matrices. However, conventional SPE sorbents often lack selectivity, leading to co-extraction of interferences, which negatively affects the following detection method. One of the advantages of MISPE is the built-in selectivity for a target analyte, or class of structurally related analytes, enabling the efficient clean-up that is often required for biological samples. The built-in selectivity of MISPE originates from the preparation of a highly crosslinked copolymer network in the presence of an imprint molecule, i.e. the template. Subsequent removal of this template molecule leads to the creation of defined recognition sites, complementary to the shape and functionality of the template. In this work, molecularly imprinted polymers were synthesized for the first time for several types of target analytes, including diphosphate esters (Papers I-III) and a protein adduct (Paper IV) and evaluated as sorbents for solid-phase extraction. A MISPE method for extracting local anaesthetic drugs from human plasma was also evaluated (Paper V). The development of appropriate methods for using the prepared polymers to extract target analytes directly from body fluids, and the elucidation of factors that influence their performance, were major foci of all the work underlying this thesis. These are not straightforward tasks, since the recognition mechanism of the material is often based on polar interactions, which are not favoured in aqueous environments. In such cases, non-selective adsorption of the analyte(s) to the polymer surface often occurs. In order to use the MIP sorbent most effectively it is important to suppress this non-selective adsorption, without disrupting the selective adsorption of the target analyte(s) to the imprints. Generally in these studies, this strong analyte-polymer surface interaction could be repressed, and selective adsorption enhanced, by carefully optimising the conditions for washing the sorbent, in terms of organic solvent volumes, solvent polarity and the addition of an ionic modifier. The sample matrix, mainly urine, was found to strongly decrease the capacity of the MIP. Hence, this effect was further investigated. It was found that the presence of NaCl in the sample negatively affected the recovery and repeatability of the method. Furthermore, these parameters could be improved by adjusting the sample pH. It was important to control the pH of the sample, in order both to achieve selective extraction and to increase the extraction recoveries. The selectivity of MISPE for the extraction of diphosphate esters from human urine was demonstrated by comparing its performance with that of a conventional SPE sorbent, a mixed-mode-anion exchanger (MAX). Due to its efficient clean-up, MISPE generated extracts that yielded less complex ion chromatograms in subsequent LC/ESI-MS analysis than extracts from the MAX cartridge. Due to its efficient clean-up, MISPE generated extracts that yielded less complex ion chromatograms in subsequent LC/ESI-MS analysis than extracts from the MAX cartridge. Signal suppression from the interfering co-eluting compounds was detected when the MAX extracts were analysed, which was not the case for the MISPE extracts. These findings show the importance of efficient and selective sample preparation, even if a selective detector is used. Development of LC/ESI-MS methods was also an extensive component of this work (Papers I-IV). Different chromatographic conditions have been evaluated for the optimal separation and detection of the investigated compounds. Use of ion-pairing agents and suitable HPLC columns (Hypercarb and C18 Aquasil) for the acidic, polar analytes, was found to give better retention and separation than use of conventional reversed-phase columns. To improve the selectivity and detectability further, selected ion monitoring (SIM) and selected reaction monitoring (SRM) acquisition modes were used for quantification of the investigated compounds. In summary, the aim of this work was to contribute to the knowledge of the recognition mechanisms of molecularly imprinted polymers in aqueous matrices, which is important for extending the use of MISPE for several types of bioanalytical applications.

Protein microarray technology has an enormous potential for in vitro diagnostics (IVD)1. Miniaturized and parallelized immunoassays are powerful tools to measure dozens of parameters from minute amounts of sample, whilst only requiring small amounts of reagent. Protein microarrays have become well-established research tools in basic and applied research and the first diagnostic products are already released on the market. However, in order for protein microarrays to become broadly accepted tools in IVD, a number of criteria have to be fulfilled concerning robustness and automation. Robustness and automation are key demands to improve assay performance and reliability of multiplexed assays, and to minimize the time of analysis. These key demands are addressed in this thesis and novel methods and techniques concerning assay automation, array fabrication as well as performance and detection strategies related to protein microarrays are presented and discussed. In the first paper an automated assay format, based on planar protein microarrays is described and evaluated by the detection of several auto-antibodies from human serum and by quantification of matrix metalloproteases present in plasma. Diffusion-rate limited solid phase reactions were enhanced by microagitation, using the surface acoustic wave technology, resulting in a slightly increased signal-to-noise ratio. In the second paper of the thesis, a novel multiplexed immunoassay system was developed by combining a direct immunoassay with a competitive system. This set-up allows quantification of analytes present in widely varying concentrations within a single multiplex assay. In the third paper, a new concept for sample deposition is introduced, addressing contemporary problems of contact or non-contact microarrayers in protein microarray fabrication. In the fourth paper, a magnetic bead-based detection method for protein microarrays is described as a cost-effective alternative approach to the commonly used fluorescence-based confocal scanning systems. The magnetic bead-based detection could easily be performed by using an ordinary flatbed scanner. In addition, applying magnetic force to the magnetic bead-based detection approach enables to run the detection step more rapidly. Finally, in paper five, a microfluidic bead-based immunoassay for multiplexed detection of receptor tyrosine kinases in breast cancer tissue is presented. Since the assay is performed inside a capillary, the amounts of sample and reagent material could be reduced by a factor of 30 or more when compared with the current standard protein microarray assay. / QC 20101112