Measurements of urinary kinins by HPLC-radioimmunoassay.
In this paper we describe a method for the individual determination of urinary kinins. Extraction from the urine is performed on an Amberlite CG-50 column and kinins are eluted with formic acid. The samples are further purified and kinins are separated by reversed phase HPLC. Bradykinin and lysylbradykinin are quantified by a sensitive radioimmunoassay capable of detecting 0.1 fmol of either peptide. Procedural losses are monitored by measuring the recovery of [3H]bradykinin and [3H]lysylbradykinin. Simple methods for labeling of bradykinin and lysylbradykinin with tritium are also presented. Recoveries of [3H]bradykinin and [3H]lysylbradykinin from biological material ranged between 77 and 91%. The combination of HPLC with radioimmunoassay makes it possible to determine kinin concentrations of biological samples with a higher sensitivity and greater specificity than previous methods.
D000818:Animals; D001920:Bradykinin / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D007609:Kallidin / Q000652:urine; D007610:Kallikreins / Q000378:metabolism; D007668:Kidney / Q000378:metabolism; D007700:Kinetics; D007705:Kinins / Q000302:isolation & purification / Q000378:metabolism / Q000652:urine*; D011863:Radioimmunoassay / Q000379:methods; D051381:Rats; D012015:Reference Standards; D013045:Species Specificity; D014316:Tritium
D007705:Kinins; D014316:Tritium; D007609:Kallidin; D007610:Kallikreins; D001920:Bradykinin