Simultaneous determination of cucurbitacin B and cucurbitacin E in rat plasma by UHPLC-MS/MS: A pharmacokinetics study after oral administration of cucurbitacin tablets

- It is the first time to simultaneous determination the pharmacokinetics of CuB and CuE in rat plasma after oral administration of cucurbitacin tablets, which have been used to treatment of chronic hepatitis.
- A simplicity and sensitivity UHPLC-MS/MS method was developed and validated.
- A better separation was obtained from the mobile phase without formic acid.
- The precision and accuracy of this method at LLOQ are within acceptable limits.

Cucurbitacin B (CuB) and cucurbitacin E (CuE) are tetracyclic triterpene compounds from Cucurbitaceae, and the main bioactive compounds of cucurbitacins tablets that used to treatment of chronic hepatitis. Pharmacological research has been very comprehensive, and there are few studies on pharmacokinetics, especially about CuE. An Ultra High Performance Liquid Chromatography-tandem Mass Spectrometry (UHPLC-MS/MS) method with high selectivity, simplicity and sensitivity has been used for quantitative analysis of Cucurbitacin B (CuB) and cucurbitacin E (CuE). Plasma samples were pretreatment by Liquid-liquid extraction (LLE) method with dichloromethane. The chromatographic separation was achieved on a C18 column (Agilent Eclipse Plus, 1.8 μm, 50 × 2.1 mm) using gradient elution with water - methanol at a flow rate of 0.3 mL/min and the column temperature was set at 30 °C. The method was validated according to FDA guidelines. Lower limit of quantification (LLOQ) was 1.60 ng/mL for CuB and 1.58 ng/mL for CuE. Correlation coefficients of CuB and CuE were more than 0.99 in rat plasma. All values of intra-day and inter-day precision (RSD%) were not exceeded 15%, the accuracy (RE%) were within −5.57 to 5.20% for CuB and −3.33 to 7.37% for CuE. The mean extraction recoveries were more than 80%. Pharmacokinetic parameters were also evaluated by UHPLC-MS/MS method. The results suggestion that this method was successfully applied to pharmacokinetic study of CuB and CuE in rat plasma after oral administration cucurbitacin tablets.