Abstract
Ursolic acid (UA) is a naturally occurring pentacyclic triterpenoid that exhibits extensive pre-systemic metabolism from in vitro studies. However, there are no available authentic metabolite standards or validated analytical methods to quantitate UA metabolites. We have identified ursolic acid sulfate (UAS) as one of the major metabolites. We were able to identify and characterize its structure via comparison to the chemically synthesized UAS. A cyano (CN, 150 × 4.6 mm, 5 µm) column along with a gradient elution of acetonitrile and 0.08% (v/v) acetic acid, pH 3.0 were employed for chromatographic separation. Negative single ion recording mode (SIR) with electron-spray ionization (ESI) source at mass-to-charge ratios of 455.3 and 535.3 were monitored for UA and UAS, respectively. UAS linearity range was 0.010–2.500 µM. The absolute values of intra-day and inter-day precision (CV, %) and accuracy (DFN, %) were all below 15%. Thus, the analytical method has been validated in the human subcellular fractions to facilitate in vitro/ in vivo DMPK and future clinical disposition studies on UA.
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