Simultaneous determination of 3,4-dihydroxyphenylacetic acid and homovanillic acid using high performance liquid chromatography-fluorescence detection and application to rat kidney microdialysate.

We established a sensitive and simultaneous determination method of 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) using HPLC-fluorescence detection. This method adopted the column-switching system, which included an on-line extraction of carboxylic acids by a strong anion-exchange column followed by separation on an ODS column, coulometric oxidation, fluorogenic reaction with ethylenediamine, and fluorescence detection. The detection limits were 50 and 100 fmol/injection for DOPAC and HVA, respectively (a signal-to-noise ratio of 3). The method was applicable to 50 microl of rat kidney microdialysate with a sufficient accuracy and precision. The concentrations of DOPAC and HVA in rat kidney microdialysate were 131+/-29 and 404+/-44 nM, respectively (n=5). This is the first report of DOPAC and HVA quantified in rat kidney microdialysate.