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Quantification of retinoyl-beta-glucuronides in rat urine by reversed-phase high-performance liquid chromatography
S Li1, A B Barua, C A Huselton
1Department of Drug Metabolism and Pharmacokinetics, Hoffmann-LaRoche, Inc., Nutley, NJ 07110, USA.
Abstract:
A method is presented for the quantitation of the glucuronide conjugates of 4-oxo-all-trans-, 4-oxo-13-cis-, 13-cis-, 9-cis- and all-trans-retinoic acids in rat urine utilizing solid-phase extraction and gradient reversed-phase HPLC. The range of the R.S.D. (relative standard deviation) for both the inter- and intra-assay precision was 1.45-11.60%. The recovery of all retinoyl-beta-glucuronides from rat urine ranged between 89 and 99%. The limit of detection was 0.01 microgram/ml using 5 ml of rat urine. This method was applied to quantitate the amount of retinoyl-beta-glucuronides produced in urine after the single and multiple oral administration of 13-cis-, 9-cis- and all-trans-retinoic acids to rats.