We will develop efficient, extremely sensitive methods to detect and screen for fatty acid amides in biological samples. We will complement the high efficiency screening, which will have limited structural information, with newly developed standardized methods for lipid profiling. The profiles will include, for the first time, fatty acid amides in the profile strategy. Using these methods, we will screen standard neuroblastoma cell lines and neuro-endocrine tissue samples to determine the presence and distribution of basal (unstimulated) levels of primary fatty amides, such as oleamide. Fatty acid amides comprise a relatively unexplored class of compounds with neuromodulatory activity. Oleamide (cis-9-octadecenamide), the """"""""parent"""""""" molecule of the class, potentiates, quite specifically, specific 5-hydroxytryptamine receptors and gamma-aminobutyric acid receptors and inhibits gap junction communication. This class of compounds may have a very specific role in the etiology of affective disorders. Such disorders as depression, anxiety, and bipolar disorders affect one in six Americans. The availability of tools to study amides is important in developing a better picture of how these lipids modulate neural transmission and other processes. These compounds are relatively difficult to study, because they are chemically and spectroscopically almost """"""""invisible."""""""" Based on substantial previous work on the chromatography, electrophoresis, and the chemistry of derivatization of these and similar lipid compounds, we have proposed a series of derivatization methods that will allow us to use high efficiency capillary electrophoresis methods coupled with high sensitivity laser-induced fluorescence for detection of amides. The development of comprehensive lipid screening protocols using liquid chromatography will allow us to optimize the isolation of amides from other lipids in such a way as to maintain the overall high efficiency of the screening process. In addition, we will incorporate, for the first time, amides in the scheme of lipid profiling. We hope that this will encourage some progress towards high resolution, comprehensive lipid profiling to complement proteomic studies of complex metabolic pathways involving bioactive lipids
