Reversed-phase ion-paired HPLC of purine nucleotides from skeletal muscle, heart and brain of the goldfish, Carassius auratus L.--I. Development of the analytical method.

1. A reversed-phase ion-paired liquid chromatographic (HPLC) method was developed to measure AMP, ADP, ATP, IMP, NAD+ and NADP+ levels in white muscle, heart and brain of anoxic goldfish. 2. Mobile phase parameters of the HPLC method (concentrations of buffer, organic modifier and counter-ion, and pH) were varied to establish the optimal conditions for separation of the compounds of interest. 3. The analytical method was evaluated by calculating some relevant chromatographic parameters (reproducibility and linearity). 4. The HPLC method showed sufficient selectivity, high sensitivity and reproducibility, and excellent linearity.

Reversed-phase ion-paired HPLC of purine nucleotides from skeletal muscle, heart and brain of the goldfish, Carassius auratus L.--II. Influence of environmental anoxia on metabolite levels.

1. The type and the amounts of some (di)nucleotides in white skeletal muscle, heart and brain of anoxic goldfish were established with a reversed-phase ion-paired HPLC method. 2. Significant changes in the levels of these metabolites, as compared to controls, were observed. The most consistent change is the increase of IMP and of IMP-load (IMP/ATP + ADP + AMP) in the three tissues during anoxia. 3. Adenylate energy charges are maintained at a high level in the anoxic white muscle and the anoxic heart. Comparison of these results with those from conventional enzymatic methods for the quantification of (di)nucleotides showed, except for IMP, no significant differences.