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Abstract
In order to evaluate the bioavailability and study the pharmacokinetics of triptolide, an HPLC method was developed for the quantitative determination of this diterpenoid in plasma. The procedure for the plasma assay employed liquid - liquid extraction with chloroform followed by high speed centrifugation. The UV absorbance of the effluent was monitored at 218 nm. An internal standard (acetophenone) was used to calibrate injection and instrument reaction errors. The proposed methodology is sensitive, rapid, and reproducible. The limit of quantitation is 0.005 mg/L in plasma (0.05 mg/L in final solution) and a linear range of determination is observed over the concentration of 0.05 mg/L to 30 mg/L. The inter- and intraday coefficients of variation for the assay of triptolide in plasma were < 16.82 % at low concentration (0.005–0.076 mg/L) and < 8.05% at high concentration (0.152–5.000 mg/L). Recovery of triptolide in plasma is greater than 96.72%.
Triptolide was stable in plasma during 30 days of storage at −80°C, whereas degradation products appeared within 4 hours when it was dissolved in methanol at room temperature. The method was employed to determine the pharmacokinetics of triptolide in rat plasma.
After oral administration of a single dose of 840 μg/kg, triptolide was found to reach a peak concentration (Cmax) of 0.210 μg/mL in 19.5min. (Tmax). The AUC was 157.28 μg and the elimination half time was 50.60 min.