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

Abstract

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 C 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 (R > 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).

Determination of Gypenoside A and Gypenoside XLIX in Rat Plasma by UPLC-MS/MS and Applied to the Pharmacokinetics and Bioavailability

“…e t 1/2 of gypenoside A in rats reported in the literature was 6.247 ± 2.039 h [17], which is significantly longer than the values we reported, but this difference was likely due to the different dosage forms. However, the t 1/2 of gypenoside XLIX in rats reported in the literature (3.17 ± 1.01) h was closer to the values we calculated [15]. Based on the pharmacokinetics data, gypenoside A and gypenoside XLIX had very low oral bioavailabilities of 0.90% and 0.14%, respectively, indicating that the concentration of the drug in systemic circulation was low.…”

“…Gypenoside A and gypenoside XLIX are the two main saponins found in Gynostemma pentaphyllum [15,16] and have been isolated from the plant and puri ed by LC-MS/MS. Guo et al performed a solid-phase extraction method and developed an LC-MS/MS method to detect gypenoside XLIX in rat plasma [15]. ey then studied the rat pharmacokinetics of gypenoside XLIX after intravenous administration.…”