1,4-Benzoquinone-based electrophoretic assay for glucose oxidase.
The communication demonstrates feasibility of an enzyme microassay for glucose oxidase with 1,4-benzoquinone as an acceptor of electrons. The protocol uses the plug-plug mode of electrophoretically mediated microanalysis, with nanolitre injected volumes of enzyme and reactant solutions. The reactant and product, 1,4-benzoquinone and hydroquinone, are separated during the assay by differential binding to sulfated-beta-cyclodextrin used as additive to the phosphate buffer (pH 7) and monitored at selected wavelengths in their UV spectra. The assay covers glucose oxidase concentration from 0.01 to 0.1mgml(-1). Due to the strong UV absorbance of the both reactant and product, there is no need for use of a second enzyme (peroxidase) in the present assay.
Urban
Pawel L
PL
Department of Chemistry, University of York, Heslington, York, YO10 5DD, UK.
Goodall
David M
DM
Bergström
Edmund T
ET
Bruce
Neil C
NC
D016227:Benzoquinones / Q000737:chemistry / Q000378:metabolism*; D019411:Clinical Laboratory Techniques; D019075:Electrophoresis, Capillary / Q000379:methods*; D005947:Glucose / Q000737:chemistry; D005949:Glucose Oxidase / Q000737:chemistry / Q000378:metabolism*; D006873:Hydroquinones / Q000737:chemistry / Q000378:metabolism; D007202:Indicators and Reagents; D007700:Kinetics; D008832:Microchemistry; D013057:Spectrum Analysis / Q000379:methods*
D016227:Benzoquinones; D006873:Hydroquinones; D007202:Indicators and Reagents; C004532:quinone; D005949:Glucose Oxidase; D005947:Glucose; C031927:hydroquinone