Rapid chromatographic purification of urinary steroid glucuronates for determination of aldosterone secretion rate

Abstract

Aldosterone secretion rate (ASR) is utilized as a rapid screening procedure to detect subtle forms of hypertension. A rapid and robust chromatographic method has been developed, based on easily prepared, rigid matrices, which permit the flow of urine by suction through sequential purification columns. Both majors metabolites, aldosterone glucuronate and tetrahydroaldosterone glucuronate, are isolated in ca. 75% yield. They are hydrolyzed and quantitated by high-performance liquid chromatography—radioimmunoassay (HPLC—RIA) or, for bulk preparations, further purified by preparative chromatofocusing. These two polar conjugates are isolated in nearly pure form by HPLC on a C 2 column with a two-step gradient. An ASR determination can be completed in one to two working days; preparative-scale work takes somewhat longer.