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.
Van der Boon
J
J
State University of Leiden, Department of Animal Physiology, Gorlaeus Laboratories, The Netherlands.
de Jong
R L
RL
Van den Thillart
G E
GE
Addink
A D
AD
D000818:Animals; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006054:Goldfish; D006863:Hydrogen-Ion Concentration; D009132:Muscles / Q000737:chemistry* / Q000378:metabolism; D009206:Myocardium / Q000737:chemistry* / Q000378:metabolism; D009928:Organ Specificity; D011685:Purine Nucleotides / Q000032:analysis* / Q000378:metabolism; D012044:Regression Analysis
D011685:Purine Nucleotides