Type

Journal Article

Authors

R O'Kennedy
A J Killard
D P Bogan

Subjects

Microbiology

Topics
umbelliferones 7 hydroxycoumarin 7 hydroxycoumarin glucuronide cattle calibration curves glucuronosyltransferase chemistry animals metabolism production linear detection deoxycholic acid electrophoresis capillary standards glucuronic acid uridine diphosphate glucuronic acid

The use of capillary electrophoresis for studying the in vitro glucuronidation of 7-hydroxycoumarin. (1995)

Abstract An assay utilizing CE, has been developed for studying the in vitro metabolism of 7-hydroxycoumarin to 7-hydroxycoumarin-glucuronide. A reaction mixture containing a crude preparation of bovine uridine diphosphate (UDP) glucuronyl transferase (UDPGT) and the substrates uridine diphosphate glucuronic acid (UDPGA) and 7-hydroxycoumarin was prepared. Aliquots were removed from the reaction mixture at specific time intervals and analyzed on a 57-cm untreated silica capillary for the presence of 7-hydroxycoumarin-glucuronide. Samples were electrophoresed in a 100 mM phosphate/11 mM deoxycholic acid (sodium salt)/acetonitrile electrolyte buffer, pH 7.0, at 30 kV, with detection at 320 nm. The method allowed the determination of 7-hydroxycoumarin-glucuronide without sample cleanup, with a limit of detection of 2 micrograms/mL, and a linear detection range of 0 microgram/mL to 100 micrograms/mL. The 7-hydroxycoumarin-glucuronide, concentrations were calculated from calibration curves of standards prepared in enzyme solution. The initial rate of production of 7-hydroxycoumarin-glucuronide, in the first 70 min, was 3.1 +/- 0.13 nmol/mL/min/mg protein. The method was fast and reliable with percentage relative standard deviations for concentration of 7-hydroxycoumarin-glucuronide, over time, of less than 10%.
Collections Ireland -> Dublin City University -> Publication Type = Article
Ireland -> Dublin City University -> Subject = Physical Sciences: Chemistry
Ireland -> Dublin City University -> DCU Faculties and Centres = DCU Faculties and Schools
Ireland -> Dublin City University -> Status = Published
Ireland -> Dublin City University -> DCU Faculties and Centres = DCU Faculties and Schools: Faculty of Science and Health
Ireland -> Dublin City University -> PubMed
Ireland -> Dublin City University -> Subject = Physical Sciences
Ireland -> Dublin City University -> DCU Faculties and Centres = Research Initiatives and Centres: National Centre for Sensor Research (NCSR)
Ireland -> Dublin City University -> DCU Faculties and Centres = DCU Faculties and Schools: Faculty of Science and Health: School of Biotechnology
Ireland -> Dublin City University -> DCU Faculties and Centres = Research Initiatives and Centres

Full list of authors on original publication

R O'Kennedy, A J Killard, D P Bogan

Experts in our system

1
R O'Kennedy
Dublin City University
Total Publications: 197
 
2
Anthony J. Killard
Dublin City University
Total Publications: 73