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 simpl e HPLC procedure is described for rat tibia; the response to hemorrhagic sh ock with and without blood retransfusion is also described. With respect to the measurement of nucleotides, a number of validation criteria are met. I n the anesthetized intact rat (Normal) there was a declining gradient of th e three nucleotides, expressed as nmol per dry matter, from proximal over m iddle to distal diaphysis, with the mean ratio ATP/ADP (0.21, 0.20, 0.20) a nd 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 decre ase versus Normal of ATP, ATP/ADP and energy charge. Blood retransfusion af ter 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 describe d 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 ene rgy charge may be characteristic for long bones, pointing towards different energy metabolism; and (iii) bone is a "shock organ", reflecting blood hyp operfusion, O-2 deficiency and decreased ATP in this situation. (C) 2001 El sevier Science B.V. All rights reserved.