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 hig h-energy phosphate (HEP) compounds in a single run and inorganic phosphate (P-i) in an other short run under the same HPLC conditions. Inorganic phosp hate was estimated by using thymidine phosphorylase (EC 2.4.2.4) which cata lyzes a reaction involving inorganic phosphate to produce 2-deoxyribose 1-p hosphate and thymine. The thymine/P-i stoichiometry was 1. The method provi des a reproducible instrument fur evaluating myocardial high-energy metabol ism under physiological and pathological conditions. (C) 2001 Elsevier Scie nce B.V. All rights reserved.