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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
chemistry
High-performance liquid chromatographic enantioselective assay for the measurement of ketoprofen glucuronidation by liver microsomes
Journal of Chromatography B: Biomedical Sciences and Applications, Volume 654, No. 1, Year 1994
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Description
A stereoselective high-performance liquid chromatographic (HPLC) method was developed to study the in vitro glucuronidation of ketoprofen enantiomers by liver microsomes. The HPLC system consisted of a Superspher 100 RP 18 end-capped column eluted with a mixture of acetonitrile and 10 mM tetrabutylammonium bromide in 1 mM potassium phosphate adjusted to pH 4.3 (30:70, v/v). Ultraviolet detection was performed at a wavelength of 254 nm. The capacity factors of S-ketoprofen glucuronide, R-ketoprofen glucuronide and R,S-ketoprofen were 12.8, 14.5 and 18.1, respectively. Sample pretreatment consisted of protein precipitation in microsomal incubation suspensions and further purification on a Sep Pak C18 cartridge before injection onto the HPLC system. Quantitation was performed with standard glucuronides biosynthetized with immobilized microsomes and purified by semi-preparative HPLC. The linearity of the method between 1.25 and 25.0 μg ml-1 (coefficient of correlation greater than 0.999), the repeatability (coefficient of variation = 1.2%; n = 5), and recovery (within 85%) were tested. The limit of detection was 10 ng for each glucuronide injected. The in vitro glucuronidation of R- and S-ketoprofen was measured in liver microsomes from man and from various animal species (dog, rat, rabbit). For both enantiomers, dog presented the highest specific activity. In contrast, the lowest activity was found in rabbit. On the other hand, the formation ratio of the S- and R-glucuronides of ketoprofen was close to 1 in man, rat and rabbit, but was 4.5 in dog, thus indicating that the reaction was stereoselective in this species. © 1994.
Authors & Co-Authors
Chakir, Saïd
Morocco, Meknes
Faculté Des Sciences de Meknès
Maurice, Marie Hélène
France, Nancy
Laboratoire Cnrs Centre du Médicament, Nancy
Magdalou, Jacques
France, Nancy
Laboratoire Cnrs Centre du Médicament, Nancy
Leroy, Pierre
France, Nancy
Laboratoire Cnrs Centre du Médicament, Nancy
Dubois, Nathalie
France, Paris
Cnrs Centre National de la Recherche Scientifique
Lapicque, Françloise
France, Paris
Cnrs Centre National de la Recherche Scientifique
Zaïd, A.
Morocco, Meknes
Faculté Des Sciences de Meknès
Nicolas, Alain
France, Nancy
Laboratoire Cnrs Centre du Médicament, Nancy
Statistics
Citations: 17
Authors: 8
Affiliations: 3
Identifiers
Doi:
10.1016/0378-4347(93)E0450-5
ISSN:
03784347