AN ENZYMATIC METHOD FOR THE DETERMINATION OF ATP AND GLYCEROL WITH ANAUTOMATED FIA SYSTEM

A method for the enzymatic determination of adenosine triphosphate (AT P) and glycerol has been developed, using an automated FIA system. The enzymes Glycerophosphate Oxidase (GPO) and Glycerol Kinase (GK) have been immobilized on non-porous glass beads following optimization of t he immobilization parameters. The fabricated Single Bead String Reacto rs (SBSR) were used in connection with a FIA system, following optimiz ation of its parameters. The half-life of GK SBSR, regarding enzyme ac tivity, was found to be 45 days for 90 measurements of daily use and s torage at 4 degrees C in the appropriate buffer. The linear dynamic ra nges of the working curves for the enzymatic determinations of ATP and Glycerol, were 4-70 mu mol.l(-1) and 2-160 mu mol.l(-1) respectively. Glycerol was determined in white and red wines after a 1:5,100 diluti on. Glycerol was also determined in human serum, following deproteiniz ation of the samples by gel filtration. The low limits of quantificati on (4 mu mol.l(-1) ATP and 1 mu mol.l(-1) glycerol) and accuracies bet ter than 2% for ATP and 1% for glycerol, show that the proposed method offers many significant improvements over the conventional methods.