Determination of aldosterone secretion rate utilizing mixed mode and high performance liquid chromatography

O. Bryan Holland, Martin Risk, Helle Brown, Claude Benedict, G. Steven Hammond

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

A simpler method for determining aldosterone secretion rate (ASR) has several applications. High performance liquid chromatography (HPLC) has several advantages over traditional Chromatographie methods for purification to constant specific activity of aldosterone liberated from its 18-glucuronide by acid hydrolysis. We found it necessary to introduce several modifications to remove urochromes before HPLC. Two methods for determining ASR were developed. With Method A a more traditional initial procedure was followed, and Sephadex LH-20 chromatography allowed removal of considerable urochromes before HPLC. However, aldosterone recovery was improved with Method B, which employed several bonded phase silica derivatives (Sepralytes) and a PBE 94 column to remove urochromes before HPLC. With this procedure the Sephadex LH-20 chromatography was not required. Aldosterone purification to constant specific activity was achieved by HPLC on a diol column with a normal phase system, and quantification was performed by RIA. ASR determinations were equivalent with both methods. This methodology should be applicable to other steroid secretory rate determinations and to applications involving purification of steroid conjugates.

Original languageEnglish (US)
Pages (from-to)567-573
Number of pages7
JournalJournal of Steroid Biochemistry
Volume25
Issue number4
DOIs
StatePublished - 1986

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High performance liquid chromatography
Aldosterone
High Pressure Liquid Chromatography
Purification
Chromatography
Steroids
Glucuronides
Secretory Rate
Silicon Dioxide
Hydrolysis
Derivatives
Recovery
Acids

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology

Cite this

Determination of aldosterone secretion rate utilizing mixed mode and high performance liquid chromatography. / Bryan Holland, O.; Risk, Martin; Brown, Helle; Benedict, Claude; Hammond, G. Steven.

In: Journal of Steroid Biochemistry, Vol. 25, No. 4, 1986, p. 567-573.

Research output: Contribution to journalArticle

Bryan Holland, O. ; Risk, Martin ; Brown, Helle ; Benedict, Claude ; Hammond, G. Steven. / Determination of aldosterone secretion rate utilizing mixed mode and high performance liquid chromatography. In: Journal of Steroid Biochemistry. 1986 ; Vol. 25, No. 4. pp. 567-573.
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