Determination of high-energy phosphate compounds and inorganic phosphate by reversed-phase high-performance liquid chromatography: evaluation of myocardial metabolic status in aerobically perfused and hypoxic mouse heart.
The present paper describes a simple HPLC method designed for measuring high-energy phosphate (HEP) compounds in a single run and inorganic phosphate (Pi) in an other short run under the same HPLC conditions. Inorganic phosphate was estimated by using thymidine phosphorylase (EC 2.4.2.4) which catalyzes a reaction involving inorganic phosphate to produce 2-deoxyribose 1-phosphate and thymine. The thymine/Pi stoichiometry was 1. The method provides a reproducible instrument for evaluating myocardial high-energy metabolism under physiological and pathological conditions.
Institute of Experimental Pathology, Faculty of Medicine, University of Ancona, Italy. patsper6@popcsi.unian.it
M
M
Tomasetti
L
L
Graciotti
G
G
Fulgenzi
A L
AL
Granata
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000860:Hypoxia / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009206:Myocardium / Q000737:chemistry* / Q000201:enzymology; D009943:Organophosphorus Compounds / Q000032:analysis*; D010710:Phosphates / Q000032:analysis*; D015203:Reproducibility of Results; D013939:Thymidine Phosphorylase / Q000378:metabolism
D009943:Organophosphorus Compounds; D010710:Phosphates; D013939:Thymidine Phosphorylase