High yield-purification of a urinary Na+-pump inhibitor |
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| Authors: | J F Cloix M Crabos I W Wainer U Ruegg M Seiler P Meyer |
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| Institution: | 1. Department of Pharmacology (INSERM U7 CNRS LA 318) Necker Hospital 161 rue de Sevres 75015 Paris, France;2. Food and Drug Administration 200 C St, SW, Washington, DC, USA;3. Preclinical Research, Sandoz, Inc-4002 Basle, Switzerland;1. College of Information and Communication, National University of Defense Technology, Wuhan 430000, China;2. School of Electronic Science and Engineering, University of Electronic Science and Technology of China, Chengdu 610054, China |
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| Abstract: | A Na+-pump inhibitor was purified from 140 liters of human urine to an apparent homogeneity. Tracing of the inhibitor during the different steps of purification was achieved by simultaneous determination of its capacity to inhibit the activity of Na+,K+-ATPase and ouabain binding, and to cross-react with antidigoxin antibodies. The final purification achieved a 400,000 fold. The purification steps included flash chromatography, anionic exchange chromatography, and reversed-phase HPLC on RP18, diphenyl and phenyl packings. NMR studies indicated that the final product was a non-peptidic, possibly steroidal compound. Its molecular weight as determined by mass spectrometry was 431. |
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