Flow injection procedures with immobilized enzyme mini-columns are described for the determination of glycerol-3-phosphate, and glycerophosphorylcholine with chemiluminescent detection. The hydrogen peroxide produced on-line is coupled with a luminol (5-amino-2,3,-dihydro-1,4-phthalazinedione) perox
Determination of inorganic phosphate by flow injection method with immobilized enzymes and chemiluminescence detection
โ Scribed by Hideki Kawasaki; Katsumi Sato; Jyunko Ogawa; Yukio Hasegawa; Hidetaka Yuki
- Publisher
- Elsevier Science
- Year
- 1989
- Tongue
- English
- Weight
- 572 KB
- Volume
- 182
- Category
- Article
- ISSN
- 0003-2697
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โฆ Synopsis
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.
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