This report demonstrates a UV-embossed polymeric chip for protein separation and identification by Capillary Isoelectric Focusing (CIEF) and Matrix Assisted Laser Desportion/Ionization Mass Spectrometry (MALDI-MS). The polymeric chip has been fabricated by UV-embossing technique with high throughput; the issues in the fabrication have been addressed. In order to achieve high sensitivity of mass detection, five different types of UV curable polymer have been used as sample support to perform protein ionization in Mass Spectrometry (MS); the best results is compared to PMMA, which was the commonly used plastic chip for biomolecular separation. Experimental results show that signal from polyester is 12 times better than that of PMMA in terms of detection sensitivity. Finally, polyester chip is utilized to carry out CIEF to separate proteins, followed by MS identification. / Singapore-MIT Alliance (SMA)

In the first part of this work, magnetized nanoporous silica based microparticles (MMPs) are used for horseradish peroxidase (HRP) immobilization and applied in amperometric peroxidase-based biosensors. A homemade magnetized carbon paste electrode permits the MMPs attraction close to the electrode surface. The resulting original biosensor is applied to the investigation of enzymatic oxidation of model drug compounds namely, clozapine (CLZ) and acetaminophen (APAP) by HRP in the presence of hydrogen peroxide. The biosensor operates at a low applied potential and the signal corresponds to the electro-reduction of electroactive species enzymatically generated. The biosensor allows performing the quantitation of the two drug compounds in the micromolar concentration range. It allows also the study of thiol compounds based on the inhibition of the biosensor response. Interestingly, distinct inhibition results are observed for HRP entrapped in the silica microparticles compared to the soluble HRP. We expect that this type of biosensors holds high promise in quantitative analysis and in biotransformation studies of drug compounds. In the second part of this thesis work, HRP immobilized magnetic nanoparticles are injected on-line and magnetically retained, as a microreactor, in the capillary of a CE setup. The purpose of such a configuration is to develop an analytical tool for studying “in vitro” drug biotransformation. The advantages expected are (i) minimum sample (drug compound) and biocomponent (enzyme) consumption, (ii) high analysis throughput, (iii) selectivity and sensitivity. In order to illustrate the potential of such an instrumental configuration, it has been applied to study acetaminophen as model drug compound. The mechanistic information obtained by the HRP/H2O2 system is in agreement with literature data on acetaminophen metabolization. Horseradish peroxidase kinetic studies are realized by this setup and the apparent Michaelis constant is determined. Capillary electrophoresis permitted the identification of APAP off-line biotransformed products such as N-acetyl-p-benzoquinone imine (NAPQI), the APAP dimer and APAP polymers as inferred from literature data. The formation of the APAP dimer was further confirmed by electrospray ionization mass spectrometry.

This thesis discusses the field testing of a newly-developed product, a geosynthetic capillary break (GCB). The GCB was developed for use in engineered soil covers when a cover incorporating a capillary break effect would be desirable, but the coarse-grained material (gravel or sand) is unavailable or uneconomical. Engineered soil covers aim to reduce the amount of acid generated from sulphide bearing waste by limiting the ingress of water and/or oxygen. The GCB is a geosynthetic system that is composed of a finely ground rock flour sandwiched between two nonwoven geotextiles and manufactured as a composite layer by needle punching in a process similar to the used for GCL (geosynthetic clay liner). The goal of the GCB is to recreate the capillary break that is achieved with soil layers using a geosynthetic product that is only a few centimetres thick and that can be rolled up and for transportation, The GCB concept has been demonstrated in a previous study (Park, 2005) based on laboratory column studies and computer modelling. The goal of this project was to determine the effectiveness of the GCB when applied at field scale. Four 25 square test plots were constructed at the tailings management area (TMA) of the HudBay Minerals Inc.(HudBay) mine site located near Flin Flon, MB. One plot contained 1 m of cover soil over top of the GCB (Plot A), one contained only 1 m of cover soil (Plot B), one contained 0.3 m of cover soil over top the GCB (Plot C), and one consisted of a conventional capillary break system with 1 m of cover soil over lying 0.2 m of sand. All of the plots, along with a control plot with no cover, were instrumented with water content sensors and gas sampling ports to monitor the movement of water and oxygen through the various covers. Matric suction sensors were also installed in Plots A and B to measure the water suction within the covers. A meteorological station was installed to gather climatic data which was used to develop a water balance for each of the plots. The plots were constructed and instrumented in the fall of 2005. Data was collected and analyzed until spring of 2007. Data from the water content sensors show that the GCB was effective in increasing the water content in the soil portion of the cover system. The suction sensors show that the suction across the GCB drops significantly (40 kPa versus less than 1 kPa) as compared to plots which contain no GCB. Data from the gas concentration sensors show that the plots containing capillary breaks reduce the oxygen flux into the tailings. The plots containing the GCB (Plots A and C) resulted in the lowest flux rates, followed by the sand capillary break (Plot D )and no capillary break (Plot B), respectively. This reduction in oxygen flux will reduce the amount of acid generated from waste, as oxygen is required for the creation of acid mine drainage. Overall the study demonstrated that at field scale that the GCB is effective in limiting the ingress of water and oxygen into the tailings under the observed conditions and the manufactured GCB is comparable to the performance of the previous hand constructed column tests.

One of the major issues in the successful decommissioning of any waste disposal system is to mitigate the spread of contaminants into the surrounding environment. In many instances this is achieved by reducing amounts of net percolation and/or oxygen diffusion into the underlying waste. An engineered cover system incorporating a capillary break is a common solution to this problem. However, traditional soil capillary breaks can often be impractical for large facilities where desirable construction materials are not readily available. The primary objective of this research is to show the initial steps in the development of a new type of geosynthetic product, namely a geosynthetic capillary break (GCB). This new product, composed of a nonwoven geotextile coupled with a fine-grained rock flour, will function similar to, and has the possibility of replacing traditional, soil capillary breaks in many applications. The specific objectives of this research are to: i) determine the pertinent material parameters of the materials used to evaluate the GCB; ii) examine one-dimensional column testing of a typical engineered soil cover system incorporating the GCB; and iii) model the cover systems to better understand current performance and predict long-term performance of the GCB. The GCB was evaluated based on the objectives outlined above. The material characterization consisted of the selection of suitable materials for the GCB, as well as the determination of their unsaturated properties. The results indicate that a geotextile-rock flour combination will develop a capillary break within an engineered cover. The one-dimensional column tests evaluated four cover systems. Soil thicknesses of 30 and 60 cm were utilized, with one column of each cover thickness incorporating the GCB. The columns were tested under both high evaporative fluxes and high infiltration rates over the course of 111 days. The measured results show that there is less moisture movement in columns that incorporate the GCB. A coupled soil-atmospheric finite element model was then used to develop a predictive model for the cover systems. The model was calibrated to the measured results from the column testing to ensure consistency. The parameters obtained from this model were used to evaluate an engineered cover system incorporating the GCB for a minesite in Flin Flon, MB. The results from the predictive modeling show that moisture infiltration is reduced approximately 80% when comparing columns with the same cover thickness. Oxygen diffusion is also reduced by 20 to 25% with the inclusion of the GCB.

The hydraulic-mechanical behaviour of swelling clay is examined in this thesis. The study includes laboratory testing and numerical modeling which considers the influence of boundary conditions on the hydraulic-mechanical behaviour of a compacted unsaturated swelling clay soil. The laboratory testing component of this research consists of three (3) series of tests using a newly modified triaxial apparatus on which mechanical and hydraulic boundary conditions are altered during liquid infiltration. Mechanical boundary conditions range from constant volume to constant mean stress and also include constant stiffness which is a spring type boundary consisting of both volume expansion and mean stress increase. Hydraulic boundary conditions include drained and undrained flow into triaxial specimens. The numerical modeling component of this research includes the creation of a new capillary tube model for swelling clay materials and incorporates dynamic changes to the cross-sectional area for flow. Laboratory results are modeled using the capillary tube model, an empirical hydraulic model, D’Arcy’s Law, and in an elastic-plastic context for unsaturated soil. Results of the laboratory and numerical modeling components show that boundary conditions dominate the hydraulic-mechanical behaviour of unsaturated swelling clay soil during liquid infiltration. In particular, a mechanism is shown to explain how hydraulic conductivity of a swelling soil can decrease with increasing water content at constant void ratio. Finally hydraulic and mechanical behaviour cannot be considered separately in swelling materials due to the intimate relationship in their response. / October 2006

Groundwater flow in the capillary fringe is rarely measured during hydrogeological studies because of the difficulties associated with investigating this region. Previous research using a point velocity probe (PVP) to investigate groundwater velocity below the water table suggested that the PVP may also be capable of measuring groundwater velocity within the capillary fringe. The earlier PVP was redesigned for this study to allow for groundwater velocity data to be collected remotely. Using this system, groundwater velocity in the capillary fringe was investigated under field and laboratory conditions. Field experiments to investigate horizontal flow in the capillary fringe were conducted either by collecting vertical velocity profiles across the water table, or by holding the probe stationary and allowing seasonal recharge to move the capillary fringe and water table past the probe. Laboratory experiments were conducted in a controlled flow tank that simulated regions of an aquifer up to 85 cm above the water table. The redesigned PVP performed well as a remote system and provided velocity measurements up to 12 cm above the water table under field conditions. These values were consistent with those measured below the water table. In the laboratory, under conditions of drainage, groundwater velocity measurements in the capillary fringe consistent with values below the water table were measured up to 44 cm above the water table. The ability to measure horizontal flow of groundwater in the capillary fringe may open up new avenues for research in the study of contaminant transport in phreatic aquifers.

Groundwater flow in the capillary fringe is rarely measured during hydrogeological studies because of the difficulties associated with investigating this region. Previous research using a point velocity probe (PVP) to investigate groundwater velocity below the water table suggested that the PVP may also be capable of measuring groundwater velocity within the capillary fringe. The earlier PVP was redesigned for this study to allow for groundwater velocity data to be collected remotely. Using this system, groundwater velocity in the capillary fringe was investigated under field and laboratory conditions. Field experiments to investigate horizontal flow in the capillary fringe were conducted either by collecting vertical velocity profiles across the water table, or by holding the probe stationary and allowing seasonal recharge to move the capillary fringe and water table past the probe. Laboratory experiments were conducted in a controlled flow tank that simulated regions of an aquifer up to 85 cm above the water table. The redesigned PVP performed well as a remote system and provided velocity measurements up to 12 cm above the water table under field conditions. These values were consistent with those measured below the water table. In the laboratory, under conditions of drainage, groundwater velocity measurements in the capillary fringe consistent with values below the water table were measured up to 44 cm above the water table. The ability to measure horizontal flow of groundwater in the capillary fringe may open up new avenues for research in the study of contaminant transport in phreatic aquifers.

One of the major issues in the successful decommissioning of any waste disposal system is to mitigate the spread of contaminants into the surrounding environment. In many instances this is achieved by reducing amounts of net percolation and/or oxygen diffusion into the underlying waste. An engineered cover system incorporating a capillary break is a common solution to this problem. However, traditional soil capillary breaks can often be impractical for large facilities where desirable construction materials are not readily available. The primary objective of this research is to show the initial steps in the development of a new type of geosynthetic product, namely a geosynthetic capillary break (GCB). This new product, composed of a nonwoven geotextile coupled with a fine-grained rock flour, will function similar to, and has the possibility of replacing traditional, soil capillary breaks in many applications. The specific objectives of this research are to: i) determine the pertinent material parameters of the materials used to evaluate the GCB; ii) examine one-dimensional column testing of a typical engineered soil cover system incorporating the GCB; and iii) model the cover systems to better understand current performance and predict long-term performance of the GCB. The GCB was evaluated based on the objectives outlined above. The material characterization consisted of the selection of suitable materials for the GCB, as well as the determination of their unsaturated properties. The results indicate that a geotextile-rock flour combination will develop a capillary break within an engineered cover. The one-dimensional column tests evaluated four cover systems. Soil thicknesses of 30 and 60 cm were utilized, with one column of each cover thickness incorporating the GCB. The columns were tested under both high evaporative fluxes and high infiltration rates over the course of 111 days. The measured results show that there is less moisture movement in columns that incorporate the GCB. A coupled soil-atmospheric finite element model was then used to develop a predictive model for the cover systems. The model was calibrated to the measured results from the column testing to ensure consistency. The parameters obtained from this model were used to evaluate an engineered cover system incorporating the GCB for a minesite in Flin Flon, MB. The results from the predictive modeling show that moisture infiltration is reduced approximately 80% when comparing columns with the same cover thickness. Oxygen diffusion is also reduced by 20 to 25% with the inclusion of the GCB.

This thesis discusses the field testing of a newly-developed product, a geosynthetic capillary break (GCB). The GCB was developed for use in engineered soil covers when a cover incorporating a capillary break effect would be desirable, but the coarse-grained material (gravel or sand) is unavailable or uneconomical. Engineered soil covers aim to reduce the amount of acid generated from sulphide bearing waste by limiting the ingress of water and/or oxygen. The GCB is a geosynthetic system that is composed of a finely ground rock flour sandwiched between two nonwoven geotextiles and manufactured as a composite layer by needle punching in a process similar to the used for GCL (geosynthetic clay liner). The goal of the GCB is to recreate the capillary break that is achieved with soil layers using a geosynthetic product that is only a few centimetres thick and that can be rolled up and for transportation, The GCB concept has been demonstrated in a previous study (Park, 2005) based on laboratory column studies and computer modelling. The goal of this project was to determine the effectiveness of the GCB when applied at field scale. Four 25 square test plots were constructed at the tailings management area (TMA) of the HudBay Minerals Inc.(HudBay) mine site located near Flin Flon, MB. One plot contained 1 m of cover soil over top of the GCB (Plot A), one contained only 1 m of cover soil (Plot B), one contained 0.3 m of cover soil over top the GCB (Plot C), and one consisted of a conventional capillary break system with 1 m of cover soil over lying 0.2 m of sand. All of the plots, along with a control plot with no cover, were instrumented with water content sensors and gas sampling ports to monitor the movement of water and oxygen through the various covers. Matric suction sensors were also installed in Plots A and B to measure the water suction within the covers. A meteorological station was installed to gather climatic data which was used to develop a water balance for each of the plots. The plots were constructed and instrumented in the fall of 2005. Data was collected and analyzed until spring of 2007. Data from the water content sensors show that the GCB was effective in increasing the water content in the soil portion of the cover system. The suction sensors show that the suction across the GCB drops significantly (40 kPa versus less than 1 kPa) as compared to plots which contain no GCB. Data from the gas concentration sensors show that the plots containing capillary breaks reduce the oxygen flux into the tailings. The plots containing the GCB (Plots A and C) resulted in the lowest flux rates, followed by the sand capillary break (Plot D )and no capillary break (Plot B), respectively. This reduction in oxygen flux will reduce the amount of acid generated from waste, as oxygen is required for the creation of acid mine drainage. Overall the study demonstrated that at field scale that the GCB is effective in limiting the ingress of water and oxygen into the tailings under the observed conditions and the manufactured GCB is comparable to the performance of the previous hand constructed column tests.

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