Application of chromatography to the analysis of phosphate compounds in small biopsy tissue samples in different...

Biochemical analysis of tissue biopsy samples for evaluation of the phosphate compounds of metabolism has been limited to a large tissue sample size, and thus, repeated biopsies on the same animal or patient are too difficult to obtain. We report here the use of the Bessman analyzer: anion exchange chromatography followed by automatic phosphorus analysis on small tissue samples. The method described here enables the repetitive measurement of high-energy phosphate compounds (ATP, ADP, AMP, creatine phosphate (CP], inorganic phosphorus (Pi), sugar phosphorus (glucose 6-phosphate and fructose 6-phosphate), and inosine monophosphate (IMP), an indicator of adequate biopsy processing and sample preparation. The data also emphasize the importance of adequate oxygenation of the experimental animal or patient. This method is easy to apply in almost any clinically oriented research laboratory for the study of needle biopsies from human and animal tissues and permits a more convenient and complete investigation of the high-energy phosphate compounds of intermediary metabolism than do the methods of firefly luminescence or the multiple, NAD-linked enzymatic systems required for the necessary sensitivity.