Abstract
Methods for quantitative extraction and enrichment of digoxin-like immunoreactive substances (DLIS) from human cord blood plasma (CBP) using organic solvents combined with a solid state absorbent are presented. Sephadex LH-20 column chromatography is more suitable than sephadex G-25 for DLIS separation and purification from CBP extract. The elution pattern of sephadex LH-20 chromatography of deproteinized and desalted CBP shows two distinct DLIS peaks. The first peak coelutes with the steroids aldosterone, cortisol, progesterone, 17-OH-progesterone, testosterone and estradiol, while the second DLIS peak coelutes with dehydroepiandrosterone sulfate (DHEA-S). The Na+,K(+)-ATPase inhibitory activity peak partially overlaps with the first DLIS peak. Coeluted steroids could account only partially for digoxin-like immunoreactivity in the first DLIS peak; however, they did not contribute to the Na+,K(+)-ATPase inhibitory activity, whereas the second DLIS peak could be ascribed to endogenous DHEA-S.
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