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Adenosine triphosphate (ATP) is the universal energy currency in biological systems. ATP is synthesized from adenosine mono and di-phosphate substrates. The relative ratio of ATP, ADP and AMP is an important indicator of metabolic standing: the energy charge is considered a quantitative measure of energy status, and serves as a barometer of sorts, as aberrant change in EC is associated with onset of numerous pathological states. Measurement of adenosine nucleotides, therefore, represents an important and insightful diagnostic.

In the present study, an ion pair reversed phase HPLC method was developed to separate ATP, ADP and AMP from like-phosphorylated guanosine metabolic products (e.g., GTP, GDP, GMP) enabling quantification of these nucleotides from tissue extracts.