The development of a simplified assay for detection of congeners of the microcystin (MC) hepatotoxin is described that combines the extreme sensitivity of surface-enhanced laser desorption/ionization time-of-flight MS (SELDI TOF-MS) with the superior selectivity of immunoaffinity interactions. Using methods similar to those of conventional immunoassays, MC standards were captured and enriched on immunoreactive ProteinChips coated with an MC-antibody and analyzed by TOF-MS. Unlike with conventional immunoassays, individual congeners were resolved from mixed pools. Assay conditions were optimized for the quantification of MC from untreated raw pond water at concentrations as low as 0.025 microg L(-1), well below the public health relevant guideline of 1 microg L(-1).