Simultaneous Determination of Caffeic Acid, Ferulic Acid and Isoferulic Acid in Rabbit Plasma by High PerformanceLiquid Chromatography

摘要

A simple and sensitive high performance liquid chromatographic method with reverse-phase column for the simultaneous quantification of caffeic acid, ferulic acid, and isoferulic acid in rabbit plasma was developed. An ODS column (150 mm x 4.6 mm I.D., 5 mum) was used as the stationary phase and the mobile phase consisted of acetonitrile/glacial acetic acid/water (15:0.5:85, v/v, pH adjusted to 4.5). Flow-rate was 1.0 raL/min and UV absorbance was set at 321 run. One hundred microliter of plasma was used to simultaneously measure the concentrations of caffeic acid, ferulic acid, and isoferulic acid. After a direct clean-up procedure with 10% of trifluoroacetic acid, the lower limits of quantification were 0.1 mug/mL and the standard curves were found to be linear over the concentration ranges of 0.1-100 ug/mL for caffeic acid, ferulic acid, and isoferulic acid. The average recoveries for caffeic acid, ferulic acid, and isoferulic acid were 96.2%, 98.0%, and 98.3%, respectively. The results for all compounds also showed good stability after three freeze/thaw cycles and storage at ambient temperature and 4degC for 24 hr. The assay method was successfully applied to the pharmacokinetic study of caffeic acid, ferulic acid, and isoferulic acid after an intravenous administration of caffeic acid to rabbits.