UPLC-MS/MS method for determination of avicularin in rat plasma and its application to a pharmacokinetic study

J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Aug 15:965:107-11. doi: 10.1016/j.jchromb.2014.06.015. Epub 2014 Jun 20.

Abstract

A rapid, sensitive and selective ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was developed and validated for the determination and pharmacokinetic investigation of avicularin in rat plasma. Sample preparation was accomplished through a simple one-step deproteinization procedure with 0.2 mL of acetonitrile-methanol (9:1, v/v) to a 0.1 mL plasma sample. Plasma samples were separated by UPLC on an Acquity UPLC BEH C18 column using a mobile phase consisting of acetonitrile-0.1% formic acid in water with gradient elution. The total run time was 1.60 min and the elution of avicularin was at 1.20 min. The detection was performed on a triple quadrupole tandem mass spectrometer in the multiple reaction-monitoring (MRM) mode using the respective transitions m/z 434.1→301.3 for avicularin and m/z 237.2→194.3 for carbamazepine (IS), respectively. The calibration curve was linear over the range of 10-3000 ng/mL with a lower limit of quantitation (LLOQ) of 10 ng/mL. Mean recovery of avicularin in plasma was in the range of 84.2-89.5%. Intra-day and inter-day precision were both <12%. This method was successfully applied in pharmacokinetic study after intravenous administration of 5.0mg/kg avicularin in rats.

Keywords: Avicularin; Pharmacokinetics; Rat plasma; UPLC–MS/MS.

MeSH terms

  • Animals
  • Chromatography, High Pressure Liquid / methods*
  • Drug Stability
  • Flavonoids / blood*
  • Flavonoids / chemistry
  • Flavonoids / pharmacokinetics
  • Limit of Detection
  • Liquid-Liquid Extraction
  • Male
  • Rats
  • Rats, Sprague-Dawley
  • Reproducibility of Results
  • Tandem Mass Spectrometry / methods*

Substances

  • Flavonoids
  • avicularin