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Clinical Chemistry 43: 715-722, 1997;
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(Clinical Chemistry. 1997;43:715-722.)
© 1997 American Association for Clinical Chemistry, Inc.

Articles

Development of enzyme immunoassay for endogenous ouabain-like compound in human plasma

Steven Harwood1, John A. Little3, Gerard Gallacher1, David Perrett2, Raymond Edwards3 and Anne Dawnay1,a

1 Department of Clinical Biochemistry and
2 Department of Medicine, Renal Research Laboratory, St. Bartholomew's Hospital Centre for Clinical Research, London ECIA 7BE, UK.

3 Netria, Department of Chemical Endocrinology, St. Bartholomew's Hospital, London EC1A 7BE, UK.
a Author for correspondence. Fax 44-171-796-4676; e-mail A.B.Dawnay{at}mds.qmw.ac.uk

Widespread evidence supports the existence of an endogenous digitalis-like compound in mammals. We report here the development of a novel enzyme immunoassay for ouabain that, in conjunction with a detailed HPLC study, identifies a ouabain-like compound (OLC) in extracted human plasma. The assay is sensitive—minimum detection limit for OLC 37 pmol/L (11 pmol/L in plasma)—and has a working range (between-assay CV <10%) of 180–10 000 pmol/L (54–3000 pmol/L in plasma). Mean recoveries of ouabain added to plasma ranged from 90% to 100%, and plasma extracts diluted in parallel to the standard curve. Plasma OLC concentrations in 10 healthy volunteers averaged 92 pmol/L (range 55–168), assuming 100% cross-reactivity of OLC in the ouabain assay. HPLC analysis with two distinct chromatographic conditions demonstrated that endogenous human plasma OLC co-eluted with authentic ouabain. The enzyme immunoassay is rapid and easy to perform and will support further investigation of the nature of this controversial endogenous steroid.


Key Words: indexing terms: chromatography, reversed-phase • antibody specificity




The following articles in journals at HighWire Press have cited this article:


Home page
 Clin. Chem.Home page
O. Vakkuri, S. S. Arnason, P. Joensuu, J. Jalonen, O. Vuolteenaho, and J. Leppaluoto
Radioiodinated Tyrosyl-Ouabain and Measurement of a Circulating Ouabain-like Compound
Clin. Chem., January 1, 2001; 47(1): 95 - 101.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
H. M. A. M. Qazzaz, M. A. El-Masri, and R. Valdes Jr.
Secretion of a Lactone-Hydrogenated Ouabain-Like Effector of Sodium, Potassium-Adenosine Triphosphatase Activity by Adrenal Cells
Endocrinology, September 1, 2000; 141(9): 3200 - 3209.
[Abstract] [Full Text] [PDF]


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