Determination of l-glutamate and l-glutamine by flow-injection analysis and chemiluminescence detection: comparison of an enzyme column and enzyme membrate sensor
| Autor: | Frank Preuschoff, Uwe Spohn, Maria-Regina Kula, Gert Blankenstein, Karl-Heinz Mohr |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Flow injection analysis
Chromatography biology Immobilized enzyme Chemistry Glutaminase Biochemistry Horseradish peroxidase Analytical Chemistry law.invention chemistry.chemical_compound law biology.protein Environmental Chemistry L-glutamate oxidase Hydrogen peroxide Spectroscopy Chemiluminescence Peroxidase |
| Zdroj: | Analytica Chimica Acta. 271:231-237 |
| ISSN: | 0003-2670 |
| DOI: | 10.1016/0003-2670(93)80050-u |
| Popis: | Glutamate and glutamine were determined by liminol chemiluminescence with flow-injection analysis (FIA) based on immobilized l -glutamate oxidase and glutaminase coupled with peroxidase. The laboratory-made flow-through cell of the detector has a measured volume of only 15 μl. The hydrogen peroxide produced in the first reaction is detected by luminol chemiluminescence catalysed by peroxidase. A membrane sensor and enzyme reactor based on immobilized hydrogen peroxidase are used for the determination of hydrogen peroxide. It was observed that Arthromyces ramosus peroxidase produced a 100 times stronger luminescence signal than horseradish peroxidase. By immobilization of the microbial peroxidase on a membrane inside the flow cell, simplification could be achieved with regard to apparatus, reagents and operation. The sensitivity of detection was considerably improved. In addition, the concept of a hydrogen peroxide biosensor was realized. The membrane sensor shows a detection limit of 1 × 10 −7 M for l -glutamate and 1 × 10 −6 −2.5 × 10 −3 M for l -glutamine. The calibration graphs were approximately linear in the range of 1 × 10 −7 −6 × 10 −5 for M l -glutamate and 1 × 10 −3 M for l -glutamine. The membrane sensor was stable over a period of 10 weeks ( > 1000 analyses). |
| Databáze: | OpenAIRE |
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