Simultaneous quantification of trans-resveratrol and its sulfate and glucuronide metabolites in rat tissues by stable isotope-dilution UPLC–MS/MS analysis

• A rapid, sensitive, selective UPLC–MS/MS with isotope-dilution had been developed.
• The isotope-dilution UPLC–MS/MS analysis is accurate and reliable at the nM range.
• The distribution of Resv and its metabolites in rats was studied.
• Concentrations of Resv and its metabolites were the highest in kidney.
• Heart tissue contained the most abundant unmodified Resv at 0.5 h post IV.

A rapid, sensitive, and selective ultra performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) in the multiple reaction monitoring (MRM) mode has been developed and validated to investigate the distribution of trans-Resveratrol (Resv) and its metabolites in rats following intravenous (IV) administration at 20 mg/kg body weight (BW). Resv and Resv metabolites were analyzed in the negative electrospray ionization mode and eluted with retention times of about 0.69–2.22 min with a runtime of 7 min. Stable deuterium-labeled Resv and its metabolites were used as the internal standards to correct for matrix effects and to allow for accurate quantification of Resv and its metabolites in a complex biological system. The method was validated with respect to linearity, within- and between-day precision, limit of quantification, recovery, and accuracy for all analytes. Upon examination at 0.5, 1, 2, and 4 h post-administration, concentrations of Resv and its metabolites were the highest in the kidney, followed by plasma; specifically, the glucuronidated forms were the most abundant. In the liver and the brain, the predominant forms were the sulfated derivatives. In contrast, heart tissue contained the highest concentration of unmodified Resv at 0.5 h post IV administration. The combined use of UPLC–MS/MS and isotope-dilution analysis, proved to be accurate and reliable in identifying and quantifying Resv and its various metabolites in biological samples at the nanomolar range. This technology is potentially applicable for other pharmacokinetic studies.

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