Determination of inorganic phosphate by flow injection method with immobilized enzymes and chemiluminescence detection.
A flow injection method with chemiluminescence detection has been developed for the enzymatic determination of inorganic phosphate. Purine nucleoside phosphorylase, xanthine oxidase, and urate oxidase were immobilized on controlled-pore glass beads. Hydrogen peroxide released by the enzymatic reactions of phosphate and inosine in carrier buffer was detected by the luminol-microperoxidase system in a flow cell. The calibration graph was linear over the range of 5 to 250 pmol, and reproducibility was 1.75% at 10 pmol. The detection limit was 500 fmol of phosphate in 50 microliters of sample injected. The phosphate content in deoxyribonucleic acid was measured by this method.
Kawasaki
H
H
Department of Clinical Chemistry, School of Pharmaceutical Sciences, Toho University, Chiba, Japan.
Sato
K
K
Ogawa
J
J
Hasegawa
Y
Y
Yuki
H
H
D000818:Animals; D001671:Biochemistry / Q000379:methods; D002138:Calibration; D002417:Cattle; D004247:DNA / Q000032:analysis; D004800:Enzymes, Immobilized / Q000378:metabolism*; D008163:Luminescent Measurements; D008165:Luminol / Q000378:metabolism; D010544:Peroxidases / Q000378:metabolism; D010710:Phosphates / Q000032:analysis*; D011683:Purine-Nucleoside Phosphorylase / Q000378:metabolism; D015203:Reproducibility of Results; D014503:Urate Oxidase / Q000378:metabolism; D014969:Xanthine Oxidase / Q000378:metabolism
D004800:Enzymes, Immobilized; D010710:Phosphates; D008165:Luminol; D004247:DNA; D010544:Peroxidases; C021799:microperoxidase; D014969:Xanthine Oxidase; D014503:Urate Oxidase; D011683:Purine-Nucleoside Phosphorylase