High-performance liquid chromatographic quantification of 4-methylumbelliferyl-beta-D-glucuronide as a probe for human beta-glucuronidase activity in tissue homogenates.
An internally standardized HPLC method to determine the concentration of 4-methylumbelliferone liberated from 4-methylumbelliferyl-beta-D-glucuronide by human beta-glucuronidase was developed. The assay allows the precise and rapid measurement of specific enzyme activity in human tissue homogenates. Without prior extraction the incubation mixture can be separated using a C8 column followed by fluorescence detection. The assay showed good accuracy and precision with a detection limit of 20 nM and a limit of quantification of 167 nM. The suitability of the method was shown in enzyme kinetic experiments with human liver homogenates.
Sperker
B
B
Dr. Margarete Fischer-Bosch Institute of Clinical Pharmacology, Stuttgart, Germany.
Schick
M
M
Kroemer
H K
HK
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005966:Glucuronidase / Q000032:analysis*; D006801:Humans; D006923:Hymecromone / Q000031:analogs & derivatives* / Q000378:metabolism; D008099:Liver / Q000201:enzymology
D006923:Hymecromone; C034888:4-methylumbelliferyl glucuronide; D005966:Glucuronidase