Rapid on-line microdialysis hyphenated technique for the dynamic monitoring of extracellular pyruvate, lactic acid and ascorbic acid during cerebral ischemia.

Rapid on-line microdialysis coupled with liquid chromatography was developed for the continuous monitoring of brain neurochemicals during cerebral ischemia. Isocratic separation of these analytes was achieved within 3 min, hence, over 80 analyses could be performed in a 4-h experiment. The dead volume of the microdialysis system was estimated to be less than 10 microl. The detection limits of the present assay, at a signal-to-noise ratio of five, were 2.0, 0.2 and 0.5 microM, for lactic acid, pyruvate and ascorbic acid, respectively. To validate this assay, a transient ischemia was produced by occlusion of two common carotid arteries for 10 min in an anesthetized gerbil. A microdialysis probe was inserted into the striatum of the gerbil to simultaneously monitor pyruvate, lactic acid and ascorbic acid during cerebral ischemia. Significant and dynamic changes in these analytes could be resolved in 3-min intervals. This rapid assay can be used as a tool to study dynamic changes in neurochemicals of the brain, such as during cerebral ischemia.