Abstract
A sensitive and efficient liquid chromatography-mass spectrometry (LC-MS) method was developed and validated for the determination of p-coumaric acid (CA) in rat plasma. After addition of the internal standard (IS) hydrochlorothiazide and acidification with 2 M hydrochloric acid, plasma samples were extracted by ethyl acetate and separated on a Kromasil C18 column (200 mm × 4.6 mm, 5 μm) using a mobile phase composed of methanol-0.5‰ acetic acid (60:40, v/v) within a runtime of 6.0 min. Analysis was performed in selected ion monitoring (SIM) mode with a negative electrospray ionization (ESI) interface. The target ions were m/z 163.15 for CA and m/z 295.95 for IS. The linear range was 0.01-15 μg·mL(-1), and the lower limit of quantification (LLOQ) was 0.01 μg·mL(-1). The intraday and interday precision (RSD %) were lower than 10% and accuracy (RE%) ranged from 97.1 to 103.2%. The validated method was successfully applied to the comparative pharmacokinetic study of CA in rat plasma after oral administration of CA and freeze-dried red wine, respectively. It was found that both AUC and T1/2 of CA in freeze-dried red wine were increased significantly (p < 0.05) compared with that in monomer. In addition, a double-peak profile could be observed from the concentration-time curve after oral administration of freeze-dried red wine.
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