Biospecific thermal analyzer coupled with a flowthrough immobilized enzyme column

A thermal analyzer consisting of a pair of glass thermistors was coupled with a flowthrough immobilized enzyme reactor for the determination of ATP concentration. Hexokinase was covalently bound to poly(chloromethyl-styrene-divinyl benzene) beads and packed in the reactor. The thermal analyzer was designed so as to respond to heat generation in the reactor. A maximum temperature change was exhibited within 1.5 min after the injection of a sample solution containing ATP together with glucose. Thermal decay occurred within 1 min. The thermal analyzer responded reproducibly to ATP at a constant concentration. Assays could be repeated at 3-min intervals. The effects of sample volume and flow rate on the response were examined with regard to an empirical equation. Concentration of ATP was determined with the thermal analyzer in the range of 0-10 mM. The thermal analyzer may be a simple device for the rapid determination of ATP concentration.