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Clinical Chemistry 21: 1245-1252, 1975;
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Clinical Chemistry, Vol 21, 1245-1252, Copyright © 1975 by the American Association for Clinical Chemistry

An Analytical System for Rapid Separation of Tissue Nucleotides at Low Pressures on Conventional Anion Exchangers

Joseph X. Khym 1

1 Biology Division, Oak Ridge National Laboratory, Oak Ridge, Tenn. 37830.

An analytical anion-exchange procedure has been developed for the rapid separation of acid-soluble nucleotides (the so-called "free" or tissue nucleotides). It permits assay at low pressures (275-415 kPa; 40-60 psi) in less than 1 h on 10-cm columns of Aminex resins (conventional styrene-type anion exchangers) with alkaline citrate solutions as the eluent. Separation variables have been investigated, to determine optimum conditions for the routine analysis of samples containing tissue nucleotides. Also described here is a simple solvent-extraction procedure for removing HClO4 or CCl3CO2H quantitatively from cell extracts that contain acid-soluble nucleotides: they are removed from aqueous acid solutions with a water-insoluble amine dissolved in a water-immiscible solvent.


Key Words: quantitative separation of acid-soluble nucleotides • rapid chromatographic separation of tissue nucleotides at low pressures • analysis of "free" nucleotides • removal of trichloroacetic or perchloric acids by simple extraction

Submitted on March 20, 1975
Accepted on April 8, 1975




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