Dammar-20(22)E,24-Diene-3β,6α,12β-Triol (YNPT2), as one of the main pharmacological and active components of Panax ginseng, promotes ubiquitination and degradation of
hypoxia inducible factor Ia through
proteasome, which reduces the content of
hypoxia inducible factor Ia in
tumor cells. Therefore, it is widely used in
tumor inhibition. A sensitive and specific bioanalytical method of liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the quantification of YNPT2 rat plasma has been developed.
Buspirone was used as the internal standard (IS). A 50 μl aliquot of rat plasma sample was deproteinized by 150 μl
methanol-
acetonitrile (1:1,v:v), vortex-mixed for 1 min and centrifuged at 15,000 r/min for 10 min at 4 °C. Then, 120 μl of supernatant was pipetted out into the autosampler vials and analyzed by LC-MS/MS with 10 μl injection volume. Chromatographic separation was performed on an Agilent ZORBAX XDB-C18 column (2.1 × 50 mm, 3.5 µm) with mobile phases consisting of water containing 5 mM
ammonium acetate (mobile phase A) and
acetonitrile (mobile phase B) at a flow rate of 0.6 ml/min over a total run time of 4.0 min. YNPT2 and
buspirone (IS) were detected and quantified using positive electrospray ionization in multiple reaction monitoring (MRM) mode with transitions of m/z 441.4 → 109.1 for YNPT2 and m/z 386.3 → 122.1 for IS. The linear range was 5-2000 ng/ml with the square regression coefficient (r2) of 0.9972, and the lower limit of quantification (LLOQ) was 5 ng/ml. The intra-day and inter-day precision deviations of YNPT2 ranged from 3.8 to 6.9% and 3.5-5.8%, and accuracy error ranged from -7.4-5.9% and -9.2-11.9%. The average extraction recovery of YNPT2 in rat plasma was between the range of 98.5%-102.7%. This method was successfully applied to study the pharmacokinetics of YNPT2 in rats after intragastric administration at a single dose of 10.0 mg/kg and after
intravenous injection at a single dose of 2.0 mg/kg.