Purpose: Polydatin is a natural product possessing various pharmacological effects. A few in vitro and in vivo studies have been conducted to determine polydatin’s efficacies. However, the lack of sensitive and specific analytical method for polydatin hampers the investigation of this promising compound. The purpose of this study is to develop a sensitive LC-MS method to simultaneously quantify polydatin and its metabolite resveratrol and to determine PK parameters and drug distribution of polydatin and resveratrol with oral administration of the titled compound.
Methods: A Shimadzu Ultra performance liquid chromatography (UPLC) system coupled to an AB Sciex QTrap 4000 mass spectrometer was used for the analysis. Separation was achieved using an Aquity UPLC BEH C 18 column (2.1 x 50 mm) with acetonitrile and water with 0.1% formic acid as the mobile phases. Analysis was performed under negative ionization electrospray mass spectrometer via the multiple reaction monitoring.Extraction recovery, matrix effect, precision, and stability tests were performed at low, medium and high concentration levels(20 ng/mL, 400 ng/mL, and 800 ng/mL respectively). Additionally, S9 fraction metabolic assays were preformed to confirm microbiota ability to hydrolyze the glucuronide moiety of polydatin to release resveratrol. Pharmacokinetic studies were performed to determine the PK parameters and drug distribution with the administration of polydatin.
Results: The developed method was linear in the range of 10 – 1000 ng/mL for both resveratrol and polydatinwith correlation coefficient values >0.99. The method has been shown to be reproducible, with intra- and inter-day accuracy and precision ±10.4% of nominal values, for both analytes. Theintra- and inter-day accuracy and precision of polydatin at low, medium, and high concentrations were 100.73 ± 7.86%, 97.95 ± 5.77%, and 102.03 ± 3.53%, respectively. The values for resveratrolwere101.11 ± 8.08%, 99.49 ± 9.14%, and 103.55 ± 5.11%, respectively. The average extraction recovery rates and matrix effect obtained by measuring triplicates of QC samples at low, medium and high concentration levels of polydatin and resveratrol in humanplasma are shown in the tables. The analyte mixture in plasma was found to be stable under bench-top, freeze-thaw, and storage (– 4° C) conditions. The metabolic studies showed thatpolydatin can be hydrolyzed by fecal S9 fractions prepared from gut microflora and PK studies showed that both polydatin and resveratrol were exposed in the plasma and variable tissues.
Conclusion: This novel UPLC-MS-MS method can comprehensively evaluate and quantify the levels of both polydatin and its major metabolite resveratrol in biological samples. These findings suggested that attentions should be paid regarding to polydatin’s in vivoefficacy because both polydatin and resveratrol were present with polydatin administration.
Christabel Ebuzoeme– Graduate Student, Texas Southern University, Houston, Texas
Yang Wang– Postdoctoral Fellow, University of Houston, Houston, Texas
Jing Ma– Texas Southern University, Houston, Texas
Dong Liang– PROFESSOR AND CHAIR, Texas Southern University, Houston, Texas
Song Gao– Assistant Professor, Texas Southern University, Houston, Texas