Simplified Determination of Blood Adenosine Triphosphate Using the Firefly System

Abstract

Abstract A simplified method is described for the determination of red cell ATP using the firefly lantern extract method. Variables investigated include the effect of the time of reading, dilution of firefly extract and the effective range of the method. Excellent recoveries were obtained. Optimal extraction of ATP from red cells was achieved with a hypotonic buffer at pH 9.2. The method could be used with acid-citrate-dextrose, heparin or EDTA as an anticoagulant. The method was found to be highly specific when the nucleotides found in normal human blood were investigated; only adenosine diphosphate and guanosine triphosphate gave slight readings, neither of which would significantly affect ATP determinations of human blood. Normal human values were found to be 5.45 µmoles of ATP/Gm. of hemoglobin or 1.83 µmoles/ml. red cells in heparinized blood samples. This method is believed to be more rapid, more reproducible and more accurate than any previously described method of ATP determination.