ULTRA-SENSITIVE DETECTION METHODOLOGIES FOR SUBSTANCE P

本研究提案的最终目标是开发一种测量高离子生物液体中低皮摩尔（pM）浓度的P物质（SP）及其代谢物的方法。该提案的第一个目标是开发一种预浓缩和富集方法，以检测低至亚皮摩尔水平的SP及其代谢物。这将通过评价几种不同类型的固相树脂和膜在保留生物基质中低浓度SP方面的有效性来实现。本研究的第二个目标是使用这种预浓缩/反相结合的方法与毛细管电泳（CE）与激光诱导荧光（LIF）检测操作与柱前或柱后衍生。一旦这些目标得到满足，就有可能使用脑的微透析取样来研究SP穿过血脑屏障的运输和代谢。随后的工作将比较可实现的检测限由发达的CE/LIF方法可获得的毛细管液相色谱法（CLC）与质谱（MS）检测。未来的研究将致力于建设一个包容性的微加工系统，包含预浓缩/浓缩装置，分离毛细管，和检测系统。

The ultimate goal of this research proposal is to develop a methodology for the measurement of low picomolar (pM) concentrations of substance P (SP) and its metabolites in highly ionic biological fluids. The first goal of this proposal is to develop a preconcentration and desalting method to permit the detection of low-to-subpicomolar levels of SP and its metabolites. This will be accomplished by evaluating several different types of solid phase resins and membranes for their effectiveness in retaining low concentrations of SP from biological matrixes. The second goal of this research is to use this preconcentration/desalting methodology in conjugation with capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection operating with either pre or postcolumn derivatization. Once these goals have been met, it will be possible to investigate the transport and metabolism of SP across the Blood Brain Barrier using microdialysis sampling of the brain. Later work will compare the detection limits achievable by the developed CE/LIF methodology to those obtainable by capillary liquid chromatography (CLC) with mass spectrometric (MS) detection. Future studies will work toward the construction of an inclusive microfabricated system that contains the preconcentration/desalting device, separation capillary, and detection system.