A flow injection analysis system involving immobilized NADH oxidase in column form for clinical analysis
| Autor: | Masayuki Totani, Masayoshi Tabata, Masaki Ikemoto, Takashi Murachi |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
Detection limit
Flow injection analysis Chromatography Immobilized enzyme Bioengineering Dehydrogenase General Medicine Enzymes Immobilized NAD Applied Microbiology and Biotechnology law.invention Luminol Potassium ferricyanide chemistry.chemical_compound chemistry Multienzyme Complexes law Humans Colorimetry Magnesium NADH NADPH Oxidoreductases Hydrogen peroxide Biotechnology Chemiluminescence |
| Zdroj: | Journal of Biotechnology. 14:33-41 |
| ISSN: | 0168-1656 |
| DOI: | 10.1016/0168-1656(90)90015-4 |
| Popis: | A highly sensitive FIA system for chemiluminometric determination of reduced coenzyme, NADH, was developed, using immobilized NADH oxidase from Brevibacterium ammoniagenes . The enzyme catalyzed the oxidation of NADH generating hydrogen peroxide which emitted chemiluminescence when mixed with luminol and potassium ferricyanide. The immobilized enzyme reactor was a mini-column, measuring 1 or 2 mm in inner diameter and 20 mm in length, and the sample volume was only 1 μl per assay, with a feeding speed of one sample per min and a lowest detection limit of 10 pmol NADH. A FIA system was also developed for the determination of magnesium in human serum, using an enzyme column reactor with simultaneously coimmobilized hexokinase, D-glucose-6-phosphate dehydrogenase, and NADH oxidase. The performance of the system was as satisfactory as a routine colorimetric assay, but with much higher sensitivity. |
| Databáze: | OpenAIRE |
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