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

Abstract 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).