A HPLC-MS/MS method for determination of 6′′′-feruloylspinosin in rat plasma and tissues: Pharmacokinetics and tissue distribution study

• A HPLC-MS/MS method was developed for assaying 6′′′-feruloylspinosin in rats.
• Its pharmacokinetics and tissues distribution were studied for the first time.
• Its sedative effect is through enhancing the mRNA expression of GABA receptors.

A sensitive, reliable and accurate HPLC-MS/MS method was developed and validated for the quantification of 6′′′-feruloylspinosin in rat plasma and tissues with puerarin as the internal standard. The separation was performed on a Proshell 120 EC-C18 column (4.6 × 150 mm, 2.7 μm) with a mobile phase consisting of acetonitrile and 0.1% formic acid (20:80, v/v) at 0.3 mL/min. The quantification was performed by MRM with m/z [M–H]− 783.3 → 427.2 for 6′′′-feruloylspinosin and m/z [M–H]− 415.4 → 295.4 for the internal standard, respectively. The calibration curves covered over a concentration range of 20–2000 ng/mL in plasma and various tissues samples (heart, liver, spleen, lung, kidney, stomach, intestine, muscle, cerebrum and cerebellum) with good linearity (r2 ≥ 0.9914). Both the intra- and inter-day precisions were less than 14.70%, and the accuracy (RE%) ranged from −5.80% to 4.93%. The extraction recoveries were within 75.21–92.96%, and the matrix effect ranged from 87.21% to 113.44%. Compared with spinosin, 6′′′-feruloylspinosin was distributed in rats faster whereas more slowly eliminated from the plasma. 6′′′-Feruloylspinosin could be distributed rapidly and widely in various tissues, and transfer across the blood–brain barrier. In addition, both 6′′′-feruloylspinosin and spinosin could enhance the expression of GABAAα1, GABAAα5, GABABR1 mRNA in rat hippocampal neurons significantly, indicating the bioactivity mechanism of 6′′′-feruloylspinosin was involved in the GABA receptors.

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