Extraction and measurement of myocardial nucleotides, nucleosides, and purine bases by high-performance liquid chromatography |
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| Authors: | D F Hammer D V Unverferth R E Kelley P A Harvan R A Altschuld |
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| Affiliation: | Department of Internal Medicine, Ohio State University Medical Center, Columbus 43210. |
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| Abstract: | Nucleotides, nucleosides, and purine bases were extracted from human endomyocardial biopsies, freeze-clamped rat hearts, and porcine coronary sinus plasma. Perchloric acid extracts were neutralized with Freon-trioctylamine and analyzed at 250 nm by reverse-phase ion-pairing high-performance liquid chromatography. To achieve the sensitivity necessary for analyzing small (1-3 mg wet wt) tissue samples, a small-bore, 2.1-mm-internal-diameter, C18, 5-micron reverse-phase column and a flow rate of 0.2 ml/min were used. All of the myocardial nucleotides and AMP degradation products were resolved in a total separation time of 27 min with 30 mM KH2PO4, 7.5 mM tetrabutylammonium phosphate buffers, and binary pH and acetonitrile gradients. |
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