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Clinical Chemistry 25: 235-241, 1979;
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Clinical Chemistry, Vol 25, 235-241, Copyright © 1979 by American Association for Clinical Chemistry

Reversed-phase liquid chromatographic separation of 3',5'-cyclic ribonucleotides

AM Krstulovic, RA Hartwick and PR Brown

A rapid, reversed-phase "high-performance" liquid-chromatographic separation of the five naturally occurring cyclic ribonucleotides is described. The separation, optimized for the measurement of these compounds in biological samples, is short (25 min), sensitive (50-100 pmol), and requires no sample pre-concentration steps. We also report an alternative isocratic elution mode, optimized for a rapid and selective analysis for adenosine 3',5'-cyclic phosphate. The identity of chromatographic peaks in biological extracts is confirmed by several methods: retention times, co-chromatography with the reference compounds, absorbance ratios, enzymatic peak-shift with cyclic nucleotide phosphodiesterase, and stopped-flow ultraviolet-scanning techniques.


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


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H. Youn, J. Koh, and G. P. Roberts
Two-State Allosteric Modeling Suggests Protein Equilibrium as an Integral Component for Cyclic AMP (cAMP) Specificity in the cAMP Receptor Protein of Escherichia coli
J. Bacteriol., July 1, 2008; 190(13): 4532 - 4540.
[Abstract] [Full Text] [PDF]


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Copyright © 1979 by the American Association for Clinical Chemistry.