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Isolation of a urinary digitalis-like factor indistinguishable from digoxin
1Second Department of Internal Medicine, University of Tokyo, Japan.
Biochemical and Biophysical Research Communications
|December 31, 1990
Summary
Researchers purified a digitalis-like substance from human urine. This substance, identified as digoxin, inhibits [3H] ouabain binding to red blood cells, suggesting a potential endogenous source.
Area of Science:
- Biochemistry
- Pharmacology
- Analytical Chemistry
Background:
- Endogenous digitalis-like factors (EDLFs) are implicated in cardiovascular regulation.
- Previous studies suggested the presence of EDLFs in human biological fluids, but definitive characterization remained challenging.
Purpose of the Study:
- To purify and characterize a digitalis-like factor from human urine.
- To determine if the purified factor is structurally and functionally similar to known cardiac glycosides like digoxin.
Main Methods:
- Large-scale adsorption and multiple high-performance liquid chromatography (HPLC) steps for purification.
- Inhibition assay of [3H] ouabain binding to human erythrocytes.
- Comparative analysis using HPLC, thin-layer chromatography (TLC), fast atom bombardment mass spectrometry (FAB-MS), and proton nuclear magnetic resonance (1H NMR) spectroscopy.
Main Results:
- A digitalis-like factor was purified to apparent homogeneity from human urine.
- The purified substance exhibited digoxin-like immunoreactivity.
- Chromatographic behavior (HPLC and TLC) and spectroscopic data (FAB-MS and 1H NMR) were indistinguishable from authentic digoxin.
Conclusions:
- Human urine contains a substance that is biochemically and structurally identical to digoxin.
- This finding suggests that digoxin may be an endogenous compound or a metabolite produced in the human body.