A validated LC–MS/MS method for the simultaneous determination of 20‐( S )‐protopanaxatriol and its two active metabolites in rat plasma: Application to a pharmacokinetics study

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EarlyView Article

  • Published: Oct 27, 2017
  • Author: Xin Qiu, Jiali Yang, Wei Wang, Qiuhong Wu, Hong Shao
  • Journal: Journal of Separation Science


We present a validated liquid chromatography with tandem mass spectrometry method for simultaneous determination of 20‐(S)‐protopanaxatriol and its two oxidative stereoisomeric metabolites (20S,24S)‐epoxy‐dammarane‐3,6,12,25‐tetraol (M1) and (20S,24R)‐epoxy‐dammarane‐3,6,12,25‐tetraol (M2) in rat plasma. 20‐(S)‐Protopanaxatriol, M1, and M2 were extracted with methanol and separated on an ACQUITY HSS T3 column. The mass spectrometry detection was accomplished in selected reaction monitoring mode with precursor‐to‐product ion transitions of m/z 493.4→143.1 for M1 and M2, m/z 475.4→391.3 for 20‐(S)‐protopanaxatriol, and m/z 459.4→375.3 for 20‐(S)‐protopanaxadiol (internal standard). The method showed good linearity over the concentration ranges of 1–1000 ng/mL for 20‐(S)‐protopanaxatriol and 0.5–200 ng/mL for M1 and M2, with correlation coefficients of more than 0.995. The lower limits of quantification for 20‐(S)‐protopanaxatriol, M1, and M2 were 1, 0.5, 0.5 ng/mL, respectively. The intra‐ and interday precisions (RSD, %) were less than 10.41% while the accuracy (relative error, %) ranged from –3.14 to 8.73%. Under the current conditions, M1 and M2 were completely separated within 3 min. The validated assay was successfully applied to evaluating pharmacokinetic profiles of 20‐(S)‐protopanaxatriol, M1, and M2 in rat.

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