Simultaneous determination of myocardial nucleotides, nucleosides, purine bases and creatine phosphate by ion-pair high-performance liquid chromatography.
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.
Fürst
W
W
Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, Vienna, Austria.
Hallström
S
S
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D009206:Myocardium / Q000737:chemistry* / Q000166:cytology; D010725:Phosphocreatine / Q000032:analysis*; D011684:Purine Nucleosides / Q000032:analysis*; D011685:Purine Nucleotides / Q000032:analysis*; D011687:Purines / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D014020:Tissue Extracts / Q000737:chemistry
D011684:Purine Nucleosides; D011685:Purine Nucleotides; D011687:Purines; D014020:Tissue Extracts; D010725:Phosphocreatine