A NONEQUILIBRIUM RADIOIMMUNOASSAY FOR ANGIOTENSIN-II

A method for the measurement of angiotensin II levels in dog plasma is described. The method is similar to previously published assays in that it couples gradient high-performance liquid chromatography (HPLC) with radioimmunoassay (RIA) and requires blood sample collection and processing to plasma in the presence of protease inhibitors. The unique feature of the present method is that it utilized a commerically available angiotensin II RIA run under nonequilibrium conditions. Performing the angiotensin Il RIA under nonequilibrium conditions increased RIA sensitivity to allow for a minimal detectable limit of 0.75 pg/mL, a limit of detection not achievable with current commercially available RIAs. This lower limit of detection will now allow for the measurement of circulating levels of angiotensin II. Quality control pools of dog plasma fortified with 4.59-50 pg/mL angiotensin II were assayed and analytical recoveries (ARs) and coefficients of variation (CV) of 72.2%-111% and 3.67%-19.0% were observed for the respective pools.