Potentiometric assay for acid and alkaline phosphatase
| Autor: | Dominika Ogończyk, Robert Koncki, Stanisław Głąb |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Chromatography
biology Potentiometric titration Phosphatase Inorganic chemistry Acid phosphatase Biochemistry Analytical Chemistry Ion selective electrode Monofluorophosphate chemistry.chemical_compound Hydrolysis chemistry biology.protein Environmental Chemistry Alkaline phosphatase Fluoride Spectroscopy |
| Zdroj: | Analytica Chimica Acta. 538:257-261 |
| ISSN: | 0003-2670 |
| Popis: | Simple potentiometric kinetic assay for evaluation of acid and alkaline phosphatase activity has been developed. Enzymatically catalyzed hydrolysis of monofluorophosphate, the simplest inorganic compound containing P F bond, has been investigated as the basis of the assays. Fluoride ions formed in the course of the hydrolysis of this specific substrate have been detected using conventional fluoride ion-selective electrode based on membrane made of lanthanum fluoride. The key analytical parameters necessary for sensitive and selective detection of both enzymes have been assessed. Maximal sensitivity of the assays was observed at monofluorophosphate concentration near 10 −3 M. Maximal sensitivity of acid phosphatase assay was found at pH 6.0, but pH of 4.8 is recommended to eliminate effects from alkaline phosphatase. Optimal pH for alkaline phosphatase assay is 9.0. The utility of the developed substrate-sensor system for determination of acid and alkaline phosphatase activity in human serum has been demonstrated. |
| Databáze: | OpenAIRE |
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