Direct and simultaneous quantification of ATP, ADP and AMP by 1H and 31P Nuclear Magnetic Resonance spectroscopy

ATP , ADP and AMP are energy substances with vital biological significance. Based on the structural differences, a simple, rapid and comprehensive method has been established by 1 H and 31 P Nuclear Magnetic Resonance ( 1 H-NMR and 31 P-NMR) spectroscopies. Sodium 3-(trimethylsilyl) propionate-2,2,3,3-d 4 (TMSP) and anhydrous disodium hydrogen phosphate (Na 2 HPO 4 ) were selected as internal standards for 1 H-NMR and 31 P-NMR, respectively. Those three compounds and corresponding internal standards can be easily distinguished both by 1 H-NMR and 31 P-NMR. In addition, they all have perfect linearity in a certain range: 0.1–100   mM for 1 H-NMR and 1–75   mM for 31 P-NMR. To validate the precision of this method, mixed samples of different concentrations were measured. Recovery experiments were conducted in serum (91–113% by 1 H-NMR and 89–113% by 31 P-NMR).