Analysis of ATP
A pivotal role of energy metabolism has been suggested in various neurological disorders including Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, tardive dyskinesia and epilepsy. Although sophisticated noninvasive procedures are used to determine energy status in human brain, biochemical assay of adenosine-5-triphosphate (ATP) continues to be the method of choice in experimental animal studies.
Figure. Effect of dilution of HClO4 extract on sensitivity of the test. The extract was neutralized with KOH before dilution.
The results of our study showed that the sensitivity of luciferase bioluminescence (FLB) test can be enhanced several folds by using ultraturax homogenizer, perchloric acid extraction, neutralization of acid extract followed by an optimum dilution prior to assay reaction . The luminescence signal remained stable for longer time when the test was performed in perchloric acid whereas a rapid decline in the signal intensity was observed when sterile water or Tris-EDTA were used as extractants .
Figure. Effect of different extractants on the stability of luminescence signal.
Khan HA. Bioluminometric assay of ATP in mouse brain: determinant factors for enhanced test sensitivity. J Biosci 2003; 28: 379-382.