Enzymatic fluorometric methods for the microdetermination of hexose phosphates in muscle.

Our current investigations on the chemical processes associated with muscular activity (1) require more sensitive methods for the determination of various metabolic intermediates than are currently available, and the present work is a contribution to the development of such procedures. These aim at the determination of hexose monophosphates on the one hand, fructose diphosphate and triose phosphates on the other. The methodology consists of a modification of the procedures of Slater (2), by the use of fluorometry for the determination of the oxidation of reduced diphosphopyridine nucleotide. To accomplish this with the required reliability, a number of experimental details had to be critically examined.’ As will be recalled, the Slater methods rest upon the following reactions : Enzyme Preparation A: