A rapid and sensitive LC-MS/MS method for the determination of vanillic acid in rat plasma with application to pharmacokinetic study

In this study, a rapid and sensitive ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-ESI-MS/MS) method was established and validated for the determination of vanillic acid in rat plasma. Plasma samples were prepared by protein precipitation with acetonitrile. Chromatographic separation was performed on a ZORBAX RRHD Eclipse Plus C18 column (2.1 × 100 mm, 1.8 μm) with gradient elution at a flow rate of 0.3 mL/min, using mobile phase consisting of 0.1% formic acid (A) and acetonitrile (B). Vanillic acid and caffeic acid (internal standard, IS) were quantified by multiple reaction monitoring (MRM) in negative ion mode. The method was fully validated according to the FDA guidelines. The calibration curve was linear over the range of 2-1000 ng/mL with correlation coefficient more than 0.99. The carry-over, matrix effect, extraction recovery, dilution effect, intra and inter-day precision and the accuracy were within acceptable limits. The method was then applied to pharmacokinetic study of vanillic acid in rats. After oral administration at doses of 2, 5, 10 mg/kg, the plasma concentration reached the peak of 0.42 ± 0.09, 0.73 ± 0.21, 0.92 ± 0.28 μg/mL at the time of 0.55-0.64 h, respectively. The oral bioavailability (F) was calculated as 25.3-36.2 % in rat plasma. The result provided pre-clinical information for further application of vanillic acid.PMID:34555192 | DOI:10.1002/bmc.5248
Source: Biomedical Chromatography : BMC - Category: Biomedical Science Authors: Source Type: research