Analysis and distribution of esculetin in plasma and tissues of rats after oral administration

Abstract

In this study, we developed a method to quantify esculetin (6,7-dihydroxycoumarin) in plasma and tissues using HPLC coupled with ultraviolet detection and measured the level of esculetin in rat plasma after oral administration. The calibration curve for esculetin was linear in the range of 4.8 ng/mL to 476.2 ng/mL, with a correlation coefficient (r(2)) of 0.996, a limit of detection value of 33.2 ng/mL, and a limit of quantification value of 100.6 ng/mL. Recovery rates for the 95.2 ng/mL and 190.5 ng/mL samples were 95.2% and 100.3%, within-runs and 104.8% and 101.0% between-runs, respectively. The relative standard deviation was less than 7% for both runs. In the pharmacokinetic analysis, the peak plasma esculetin level was reached 5 min after administration (Cmax=173.3 ng/mL; T1/2=45 min; AUC0 ~180 min=5,167.5 ng · min/mL). At 180 min post-administration (i.e., after euthanasia), esculetin was only detectable in the liver (30.87±11.33 ng/g) and the kidney (20.29±7.02 ng/g).

Keywords: HPLC; esculetin; pharmacokinetics; plasma.

Fig. 1

Structure of esculetin.

Fig. 2

Typical MS spectra and HPLC-UV chromatogram (A) and (+) ESI-MS/MS fragmentation ions (collision energy=18 volts) (B) of esculetin.

Fig. 3

The concentration vs. time curve of esculetin in plasma (n=9).