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

• The simultaneous detection of ATP, ADP and AMP was established by qNMR.
• This method was simple, rapid, sensitive, little sample preparation and green.
• It showed excellent linearity in the range of 0.1–100 mM and 1–75 mM, respectively.

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 1H and 31P Nuclear Magnetic Resonance (1H-NMR and 31P-NMR) spectroscopies. Sodium 3-(trimethylsilyl) propionate-2,2,3,3-d4 (TMSP) and anhydrous disodium hydrogen phosphate (Na2HPO4) were selected as internal standards for 1H-NMR and 31P-NMR, respectively. Those three compounds and corresponding internal standards can be easily distinguished both by 1H-NMR and 31P-NMR. In addition, they all have perfect linearity in a certain range: 0.1–100 mM for 1H-NMR and 1–75 mM for 31P-NMR. To validate the precision of this method, mixed samples of different concentrations were measured. Recovery experiments were conducted in serum (91–113% by 1H-NMR and 89–113% by 31P-NMR).

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