Capillary Electrophoresis An investigation of the theory and operation, and its application to environmental and DNA analyses. Kristina Schmit Douglas Jones Jordan Simon Fromholz John Jr. Yoshihara Topics of Discussion What is Capillary Electrophoresis? Theoretical Background Capillary Electrophoresis System Applications and Contributions Environmental Applications Applications to DNA Analysis Conclusion What is Capillary Electrophoresis? Analytical separation technique The “movement or migration of ions… under the influence of an electric field.” Regularly applied to inorganic, organic, environmental, pharmaceutical, and biological analyses Theoretical Background Separating trace amounts of ionic species can be a highly difficult and tedious process. The separation process depends on the ions having different migration speeds. The velocity of ion migration [m/s], is defined by:: ν = μeE Theoretical Background (cont.) The mobility indicates how fast the given ion or solute moves through a medium. q e r Electrical forces move the ion forward, and frictional forces oppose the motion. [1] The Development of CE Old methods involved using paper or gels to transport the charged species to the cathode or anode. Long analysis coupled with tedious lab time made process almost impractical. The addition of a fused silica capillary in the 1980’s created a more efficient, precise method of separation and identification. The CE System The capillary serves two functions High voltage is passed through the tube charging the molecules. The tube also allows for the flow of ions from one node to the other. Ions flow by means of an induced current on the walls of the capillary, creating an ‘electron driven pump’. Ion Flow and Analysis Applying a voltage causes the flow of metal ions on the wall of the capillary creating an electroendosmotic flow (EOF). The induced flow transports the separated ions past an ultra violet absorbance that calculates their physical properties. Capillary Electrophoresis Applications & Contributions Environmental Applications CE is an important tool for detecting and separating a wide variety of compounds. CE is used in environmental monitoring. Example: Herbicides and pesticides runoff polluting streams & groundwater. Environmental Applications CE is used in determining amounts of Inorganic and organic species in Environmental water and sediments. Example: Toxic substances methyl mercury in drinking water. CE is used in separating ions. e.g. lead contamination in ground soil. Environmental Impact Cost Efficiencies CE uses less organic solvent; less product waste = less harm to environment. Shorter analysis time; reduces Cost = more efficient. Easier, cleaner preparation. = more convenient. Applications to DNA Analysis Mutation detection Provide for future advances in diagnosis of human genetic diseases. CE is capable of analyzing both known and unknown mutations. Quicker separation technique while not sacrificing effectiveness or range of applicability. Applications to DNA Analysis (cont.) Human Genome Project Lions share of separation and analysis done by CE. Provides for rapid DNA analysis and high throughput. Capillary Array Electrophoresis What This Means Separation technique capable of separating trace amounts of ionic species. Is a cleaner, cheaper, faster and more environmentally favorable method of detecting and separating environmental contaminants. Contributions to biological sciences have provided foundation for profound future advances. Future developments will broaden range of applicability