Adenosine nucleotides in rat bone measured by ion-pair reversed-phase high-performance liquid chromatography: effect of hemorrhagic shock, with and without retransfusion of blood

The measurement of bone adenosine nucleotides (ATP, ADP, AMP) using a simple HPLC procedure is described for rat tibia; the response to hemorrhagic shock with and without blood retransfusion is also described. With respect to the measurement of nucleotides, a number of validation criteria are met. In the anesthetized intact rat (Normal) there was a declining gradient of the three nucleotides, expressed as nmol per g dry matter, from proximal over middle to distal diaphysis, with the mean ratio ATP/ADP (0.21, 0.20, 0.20) and the mean energy charge (0.34, 0.31, 0.30) being low. Irrespective of the anatomic site, hemorrhagic shock of 30-min duration evoked a further decrease versus Normal of ATP, ATP/ADP and energy charge. Blood retransfusion after shock kept nucleotides and other variables in the proximal and distal, but not the middle, diaphysis within normal limits. It was concluded that: (i) bone nucleotides are reliably measurable by HPLC, allowing the described method to be recommended for wider use in bone research and related areas; (ii) in contrast to more parenchymatous tissues, low ATP, ATP/ADP and energy charge may be characteristic for long bones, pointing towards different energy metabolism; and (iii) bone is a “shock organ”, reflecting blood hypoperfusion, O2 deficiency and decreased ATP in this situation.