The Pharmacokinetics Of Resveratrol In Rats And Its Effect On CYP1A1Enzyme Activity

Objective:â‘ To establish a liquid chromatography tandem mass spectrometry(LC-MS/MS) method for the determination of resveratrol in rat plasma.â‘¡Thedeveloped method was applied in the pharmacokinetic studies of resveratrol in rats.â‘¢To evaluate the effects of resveratrol on CYP1A1enzyme activity in rat lung andliver using substrate probe. Methods: Chosing naringenin as the internal standard, theanalysis was performed on a Hypersil GOLD C18column (150mm×2.1mm,3μm)with mobile phase of methanol/water (70:30). Isocratic chromatographic separationwas run at0.3ml/min and the column temperature was30℃. The ESI source wasoperated in negative ion scanning. In the mode of multi-reaction monitoring (MRM),ions of m/z227.1â†'143.1for resveratrol and m/z271.1â†'151.1for internal standardwere detected selectively. After the plasma sample extracted with ether, the upperorganic phase was evaporated to dryness in a stream of N2. The residue was thendissolved with the mobile phase and the supernatant was injected for analysisâ‘¡Resveratrol was administered to rats by gavage at the dose of100mg/kg, whileblood samples were collected from the orbital venous plexus of rats just before theadministration of the drug and0.083,0.167,0.25,0.5,0.75,1,1.5,2,3,4,6,8,12hthereafter. The separated plasma samples were analyzed with the established methodand the DAS software was used to calculate corresponding pharmacokineticparameters.â‘¢Preparing rat lung and liver microsomes by calcium precipitationmethod and resorufin ethyl ether was applied as the CYP1A1specificity substrate. The effects of resveratrol at different concentration levels on CYP1A1enzymaticactivity in microsomes were determined for the IC50values. Results:â‘ The assayhad a good linearity over the range of2~5000μg/L with the lower limit ofquantification (LLOQ) at2μg/L. The typical standard curve wasy=0.00362x+0.01986, R2=0.9996. The intra-and inter-day precision, accuracy, matrixeffects were in the range of allowable deviation. The stability proved to be good andrecoveries were higher than80%.â‘¡Resveratrol showed rapid absorption in rats, Tmaxwas0.236±0.034h. The value of Cmaxreached1064.996±178.272μg/L and thehalf-life was2.839±1.827h.â‘¢The IC50values of resveratrol for CYP1A1in rat lungand liver microsomes were10.10μmol/L and28.26μmol/L, respectively. Conclusion:â‘ The established LC-MS/MS method has the advantages of high specificity,sensitivity and simple sample preparation, which satisfies the pharmacokinetic studyof resveratrol in rats.â‘¡Resveratrol shows rapid absorption, low plasma concentrationin rats and has a short half-life.â‘¢Resveratrol evidently inhibits CYP1A1activity inrat lung and liver, which is associated with the mechanisms of its anti-cancer effect.