High-performance liquid chromatographic analysis of a new neuroprotective agent for ischemia-reperfusion damage, KR-31378

A high-performance liquid chromatographic method was developed for the dete rmination of a neuroprotective agent for ischemia-reperfusion damage, KR-31 378, in human plasma and urine and in rat tissue homogenates. The method in volved deproteinization of the the biological samples with 0.5 volumes of s aturated Ba(OH)(2), 0.5 volumes of 0.04 M ZnSO4 and 1 volume of acetonitril e. A 80-mul aliqout of the supernatant was injected onto a reversed-phase C 18 column. The mobile phase, 50 mM triethylamine acetate : acetonitrile : t etrahydrofuran (65:30:5, v/v/v), was run at a flow rate of 1.0 ml/min. The column effluent was mornitored by a ultraviolet detector set at 310 nm. The retention time of KR-31378 was approximately 6.5 min. The detection limits of KR-31378 in human plasma and urine and rat tissue homogenates were 0.2, 0.5 and 0.5 mug/ml, respectively. The coefficients of variation (within-da y and between-day) were below 13.6% for human plasma and urine and rat homo genates. No interferences from endogenous substances were found.