Extraction and measurement of myocardial nucleotides, nucleosides, and purine bases by high-performance liquid chromatography.
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.
Hammer
D F
DF
Department of Internal Medicine, Ohio State University Medical Center, Columbus 43210.
Unverferth
D V
DV
Kelley
R E
RE
Harvan
P A
PA
Altschuld
R A
RA
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009206:Myocardium / Q000032:analysis*; D009705:Nucleosides / Q000302:isolation & purification*; D009711:Nucleotides / Q000302:isolation & purification*; D011687:Purines / Q000302:isolation & purification*; D051381:Rats; D013552:Swine
D009705:Nucleosides; D009711:Nucleotides; D011687:Purines