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 H-1 and P-31 Nuclear Magnetic Resonance (H-1-NMR and P-31-NMR) spectroscopies. Sodium 3-(trimethylsilyl) propionate-2,2,3,3-d(4) (TMSP) and anhydrous disodium hydrogen phosphate (Na2HPO4) were selected as internal standards for H-1-NMR and P-31-NMR, respectively. Those three compounds and corresponding internal standards can be easily distinguished both by H-1-NMR and P-31-NMR. In addition, they all have perfect linearity in a certain range: 0.1-100 mM for H-1-NMR and 1-75 mM for P-31-NMR. To validate the precision of this method, mixed samples of different concentrations were measured. Recovery experiments were conducted in serum (91-113% by H-1-NMR and 89-113% by P-31-NMR). (C) 2015 Elsevier B.V. All rights reserved.