Abstract
Porous polymer monolithic (PPM) columns are employed to collect and concentrate neuronal release from invertebrate and vertebrate model systems, prior to their characterization with mass spectrometry. The monoliths are fabricated in fused-silica capillaries from lauryl methacrylate (LMA) and ethylene glycol dimethacrylate (EDMA). The binding capacities for fluorescein and for fluorescently labeled peptides are on the order of nanomoles per millimeter of length of monolith material for a capillary with an inner diameter of 200 microm. To evaluate this strategy for collecting peptides from physiological solutions, angiotensin I and insulin in artificial seawater are loaded onto, and then released from, the monoliths after a desalination rinse, resulting in femtomole limits of detection via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Positioned in the extracellular media near Aplysia californica bag cell neurons, upon electrical stimulation, these LMA-EDMA monoliths are also used to collect and concentrate peptide release, with egg-laying hormones and acidic peptide detected. In addition, the collection of several known peptides secreted from chemically stimulated mouse brain slices demonstrates their ability to collect releasates from a variety of neuronal tissues. When compared to collection approaches using individual beads placed on brain slices, the PPM capillaries offer greater binding capacity. Moreover, they maintain higher spatial resolution, compared to the larger-volume, solid-phase extraction collection strategies.