A new spectrofluorimetric method for renal renin activity: a comparative study with other methods

Abstract

A new spectrofluorimetric method for protein estimation has been developed and applied to study the renin-angiotensin system.The fluorometric reagent, 1-4-diamino-2-03-dichloro-anthraquinone, is introduced in this field and used for the first time.Renal semipurified rat renin is incubated with a synthetic substrate (N-acetyl-tetra-decapeptide) at 0 degrees C and 37 degrees C, at optimal pH, during 3 hours.The incubated mixture is studied by bioassay (BA), radioimmunoassay (RIA) and spectrofluorimetric analysis (SFA), and values obtained with these methods are compared.Blank samples for spectrofluorimetric analysis were prepared by substituting the incubated mixture with the unincubated components of the reaction.Its fluorescence values were subtracted from those of the incubated mixture.Precision and sensitivity for RIA and SFA were similar in both cases, but different in the case of BA.Renal renin activity (RRA) values for RIA (7.15 x 10-(3) nM/ml angio.I/3 h) and SFA (7.08 x 10-(3) nM/ml angio.I/h) were statistically equal (t = 1.05; p less than 0.05), while RRA values for BA (6.23 x 10-(2) nM/ml angio.I/3 h), were higher and significantly different from the statistical point of view.The graphical representation of RRA values for RIA versus SFA values gives an objective expression where RRA(RIA) = 0.78 RRA(SFA) + 0.02.