title: Determination of inorganic phosphate by flow injection method with immobilized enzymes and chemiluminescence detection.
abstract: 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.
authors.affiliation: Department of Clinical Chemistry, School of Pharmaceutical Sciences, Toho University, Chiba, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Kawasaki
authors.forename: K
authors.initials: K
authors.lastname: Sato
authors.forename: J
authors.initials: J
authors.lastname: Ogawa
authors.forename: Y
authors.initials: Y
authors.lastname: Hasegawa
authors.forename: H
authors.initials: H
authors.lastname: Yuki
mesh_terms: 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
chemical_list: D004800:Enzymes, Immobilized; D010710:Phosphates; D008165:Luminol; D004247:DNA; D010544:Peroxidases; C021799:microperoxidase; D014969:Xanthine Oxidase; D014503:Urate Oxidase; D011683:Purine-Nucleoside Phosphorylase