PubMed HealthSearch

PubMed · 7353068

Enzyme electrode for phenol.

Abstract

An enzyme electrode is described for quantitative determination of phenol at micromolar concentrations. Immobilized phenol hydroxylase is attached to the surface of a Clark oxygen electrode. The maximum rate of oxygen consumption is linearly dependent on phenol concentration over the 0.5-50 microM range. The electrode can be used for at least 150 assays without an activity loss. Readout is very rapid--within 30 sec of sample addition. The electrode response is independent of pH between pH 6.5 and 9.5. The response increases linearly with temperature in the interval 10-40 degrees C. It is necessary to incubate the enzyme electrode in a buffer containing NADPH for a few minutes before the addition of sample. This is to make the electrode response independent of the diffusion rate of this cosubstrate. This and other diffusional effects on the performance of the phenol electrode are discussed.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

K G Kjellén, H Y Neujahr. 1980. Enzyme electrode for phenol.. https://doi.org/10.1002/bit.260220205

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

Electroporation-induced damage in mammalian cell DNA.

Electroporation induced damage in the DNA of HL60 cells has been investigated by alkaline elution techniques. DNA damage is minimised by reducing the total charge applied (i.e., voltage x capacitance). Reduction of either of these electrical parameters, however, compromises the induced permeability of the cells to small molecules. The data presented concerning the effects of voltage and capacitance on DNA damage and the permeability of cells can be used to specify optimum conditions for electroporation in which DNA damage is minimised. The duration for which the current is applied can be seen to have a significant effect on the level of DNA damage. A modest temperature rise may occur when an electric charge is passed through electroporation buffer, but this event alone does not induce DNA damage in cells. The effect of voltage upon the permeability of HL60 cells to fluorescent-labelled molecules of varying molecular weight is reported.

Buffers

Measurement of intravesicular volumes by salt entrapment.

Internal volume is a very sensitive parameter of vesicle morphology. Measurement of captured volumes by solute entrapment is legitimate for most types of vesicles (Perkin, W.R. et al. (1993) Chem. Phys. Lipids 64, 197-217). In this study chloride was selected as the most convenient marker ion because the ubiquity of Cl- in physiological buffers eliminates prelabeling with exogenous markers and because minute concentrations of trapped chloride are well detectable in the presence of large extravesicular nitrate concentrations. Perfect exchange of external chloride for nitrate was shown to be accomplished by gel filtration, dialysis, or sucrose gradient flotation-but only after significant technical improvements and/or elimination of experimental pitfalls. Reliability was cross-checked by simultaneous entrapment of Cl- and K+. Diafiltration and ion exchange chromatography appeared inapplicable for exchange of extravesicular salt. When a representative variety of vesicle preparations was analyzed for internal volume (as well as for external surface and size) unexpected features of vesicle morphology were discovered. This emphasizes the genuine role of macroscopic vesicle characterization in complementing information from electron microscopy.

Buffers

The interaction of nitroaromatic drugs with aminothiols.

The effect of cysteamine and glutathione addition on the redox behaviour of metronidazole, chloramphenicol, M&B 4998, nitrofurazone, and nifuroxime has been studied by electrochemical techniques. The presence of thiol influences the redox behaviour of the nitro compound in a number of ways. In aqueous media, the single-step nitro/hydroxylamine reduction shows a decrease in current and a shift to more positive potentials, which is assigned to the thiol acting as the reducing agent, but only after the formation of the nitro radical anion. In addition, the reversible RNO/RNHOH couple is greatly diminished or removed. In a dimethylformamide/H2O solvent, the nitro radical anion can be selectively generated. The effect of thiol addition on the stability of the radical anion is strongly dependent on the drug, the identity of the thiol, and the concentration of the supporting electrolyte. The presence of thiol can result in an increase or a decrease in the lifetime of the radical with no apparent correlation with the redox couple of the nitro compound, or can act as an oxidizing agent and regenerate the original nitro compound. These disparate routes by which thiol can modify the redox characteristics of nitro compounds suggest that the traditional role of thiol as a radical scavenger needs to be extended.

Buffers