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Biomed Chromatogr. 2017 Jun;31(6). doi: 10.1002/bmc.3898. Epub 2016 Dec 14.

Development of a targeted method for quantification of gypenoside XLIX in rat plasma, using SPE and LC-MS/MS.

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The Department of Anesthesiology, Zhongshan Hospital of Dalian University, Dalian, China.
The Department of Intervention, The Second Hospital of Dalian Medical University, Dalian, China.
The Department of Ophtalmology, the Second Hospital of Dalian Medical University, Dalian, China.


A sensitive, selective and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the quantification of gypenoside XLIX, a naturally occurring gypenoside of Gynostemma pentaphyllum in rat plasma and then validated according to the US Food and Drug Administration's Guidance for Industry: Bioanalytical Method Validation. Plasma samples were prepared by a simple solid-phase extraction. Separation was performed on a Waters XBridgeTM BEH C18 chromatography column (4.6 × 50 mm, 2.5 μm) using a mobile phase of acetonitrile and water (62.5:37.5, v/v). Gypenoside XLIX and the internal standard gypenoside A were detected in the negative ion mode using selection reaction monitoring of the transitions at m/z 1045.6 → 913.5 and 897.5 → 765.4, respectively. The calibration curve was linear (R2  > 0.990) over a concentration range of 10-7500 ng/mL with the lower quantification limit of 10 ng/mL. Intra- and inter-day precision was within 8.6% and accuracy was ≤10.2%. Stability results proved that gypenoside XLIX and the IS remained stable throughout the analytical procedure. The validated LC-MS/MS method was then applied to analyze the pharmacokinetics of gypenoside XLIX after intravenous administration to rats (1.0, 2.0 and 4.0 mg/kg).


LC-MS/MS; gypenoside XLIX; pharmacokinetic study; rat plasma

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