Simultaneous determination of myocardial nucleotides, nucleosides, purine bases and creatine phosphate by ion-pair high-performance liquid chromatography. |
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Authors: | W Fürst S Hallstr?m |
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Affiliation: | Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, Vienna, Austria. |
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Abstract: | An ion-pair reversed-phase high-performance liquid chromatographic method is described for the separation and quantification of myocardial nucleotides, nucleosides, their metabolites and creatine phosphate-related compounds in a single run. Separation of a standard mixture containing 21 compounds was achieved on a 5-microns Hypersil ODS column with a 5-min isocratic elution (buffer: 0.1 M NaH2PO4, pH 5.5, containing 5.9 mM tetrabutylammonium hydrogen-sulphate) followed by a slow linear gradient to 17% acetonitrile. The method was applied to extracts of freeze-clamped rat heart tissue samples as well as to extracts of neonatal rat heart cardiomyocytes, and it provided good resolution of high-energy phosphates, including creatine phosphate, as well as of their degradation products. |
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