SELECTIVE DETERMINATION OF TRIPHOSPHONUCLEOTIDE BY A FLOW-INJECTION CHEMILUMINESCENCE METHOD

Highly selective determination of triphosphonucleotide such as adenosine triphosphate (ATP) was established by a flow injection method with chemiluminescent detection of 1,10-phenanthroline. The high selectivity is obtained by loading triphosphonuleotide such as ATP chemically to copper(II)-modified clinoptirolite (zeolite). The analyte is indirectly determined by a CL detection of copper(II) eluted from Cu-Zeo through a material exchange between Cu-Zeo and analyte. The system is comprised of three flow lines, and one of them has the material exchange column (4.5 mm i.d., 15 cm long) packed with the Cu-Zeo of 2.4 g (particle size: 0.074-0.125 mm; loading of 2 X 10(-6) mol ATP). The limits of determination by 100 mul injection method were 50 fmol to 2.0 pmol for ATP, 60 fmol to 1.0 pmol for uridine triphosphate, 50 fmol to 3.0 pmol for guanosine triphosphate and 80 fmol to 1.0 pmol for cytidine triphosphate. The relative standard deviation was 2.1 % in 5 repeated runs of 1 X 10(-9) M ATP. The selectivity was high and the CL intensity of 1 X 10(-9) M ATP alone was not interfered by coexistence of 1 X 10(-4) M other nucleotides and their derivatives. The present method was also successfully applied to determine number of bacteria in normal human urine.