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OBJECTIVE Little is known about the metabolic characteristics of trans-resveratrol, a bioactive agent with multiple beneficial effects on human health.The aim of the present study was to develop a methodology for detection and identification of transresveratrol and its metabolites in mouse plasma.METHODS Trans-resveratrol was administrated orally at a single dose of 240 mg·kg-1 to C3H mouse, and blood was collected and centrifugated.After acidified by acetic acid, methanol extract of mouse plasma was analyzed for tran-resveratrol metabolites using high performance liquid chromatography with on line photodiode array detection and multi-stage mass spectrometric detection (LC-PDA-MSn).UV-photodiode array analysis facilitated the identification of cis-and trans-isomers of resveratrol and its metabolites.Negative ion electrospray mass spectrometric analysis provided molecular weight confirmation of metabolites and tandem multi-stage mass spectrometry allowed structural information to be obtained.Agilent eclipse plus C18 (2.1 × 100 mm, 3.5 μm) served as the stationary phase.The mobile phase consisted of A (organic) methanol and B (aqueous) 10 mmol · L-1 ammonium acetate in distilled water (pH =6.0) containing 5% 2-propanol at a flow rate of 0.2 ml· min-1.RESULTS From the mass chromatograms and the PDA-HPLC chromatogram of the mouse plasma sample, we can find six peaks of M0 ([M-H]-ion m/z 227), M1 (m/z 307), M2 (m/z 403), M3 (m/z 387), M4 (m/z 483) and M5 (m/z 579).All of them have a maximum absorption at the wavelength of near 305 nm.On the basis of authentic standard, M0 (m/z 227) was confirmed to be trans-resveratrol, and the other (M1-M5) were identified as its conjugated metabolites by the following multiple reaction monitoring transitions : 307→227 (loss of a sulfate), 403→227 (loss of a glucuronic acid), 387→227 (loss of two sulfates), 483→227 (loss of one sulfate and a glucuronic acid) and 579→227 (loss of two glucuronic acids), respectively.No cis-resveratrol or phase Ⅰ metabolites were detected in mouse plasma.CONCLUSION A rapid and simple method for the analysis of trans-resveratrol and its metabolites in mouse plasma was established.According to the results of the experiment, we successfully detected and identified the metabolites of trans-resveratrol which were trans-resveratrol monosulfate (M1), trans-resveratrol monoglucuronide (M2), trans-resveratrol disulfate (M3), trans-resveratrol sulfate-glucuronide (M4) and trans-resveratrol diglucuronide (M5) in the mouse plasma.It is indicated that the in vivo major biotransformation pathways of trans-resveratrol are glucuronide and sulfate conjugation in mouse.