Development and validation of a UPLC-MS/MS method for the determination of cucurbitacin B in rat plasma and application to a pharmacokinetic study

Biomed Chromatogr. 2016 Apr;30(4):503-7. doi: 10.1002/bmc.3571. Epub 2015 Sep 2.

Abstract

Cucurbitacin B (CuB), one of the most abundant forms of cucurbitacins, is a promising natural anticancer drug candidate. Although the anticancer activity of CuB has been well demonstrated, information regarding the pharmacokinetics is limited. A rapid, selective and sensitive UPLC-MS/MS for CuB was developed and validated using hemslecin A (HeA) as internal standard (IS). Plasma samples were pre-treated by liquid-liquid extraction with dichloromethane. Separation was achieved on a reversed-phase C18 column (50 × 4.6 mm, 5 µm) at 35°C using isocratic elution with water-methanol (25:75, v/v) at a flow rate of 0.3 mL/min. The analytes were monitored by a triple quadrupole tandem mass spectrometer with positive electrospray ionization mode. The calibration curve was linear (r > 0.995) in a concentration range of 0.3-100 ng/mL with a limit of quantification of 0.3 ng/mL. Intra- and inter-day accuracy and precision were validated by percentage relative error and relative standard deviation, respectively, which were both lower than the limit of 15%. This assay was successfully applied to a pharmacokinetic study of CuB in Wistar rats.

Keywords: UPLC-MS/MS; cucurbitacin B; pharmacokinetic study.

Publication types

  • Validation Study

MeSH terms

  • Animals
  • Anti-Inflammatory Agents / blood*
  • Anti-Inflammatory Agents / isolation & purification
  • Chromatography, High Pressure Liquid / economics
  • Chromatography, High Pressure Liquid / methods*
  • Cucurbitaceae / chemistry
  • Limit of Detection
  • Liquid-Liquid Extraction / economics
  • Liquid-Liquid Extraction / methods
  • Male
  • Rats, Wistar
  • Reproducibility of Results
  • Spectrometry, Mass, Electrospray Ionization / methods
  • Tandem Mass Spectrometry / economics
  • Tandem Mass Spectrometry / methods*
  • Triterpenes / blood*
  • Triterpenes / isolation & purification

Substances

  • Anti-Inflammatory Agents
  • Triterpenes
  • cucurbitacin B