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Simultaneous determination of three iridoid glycosides of Rehmannia glutinosa in rat biological samples using a validated hydrophilic interaction–UHPLC–MS/MS method in pharmacokinetic and in vitro studies
Author(s) -
Jeong SeungHyun,
Jang JiHun,
Cho HeaYoung,
Lee YongBok
Publication year - 2020
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.202000809
Subject(s) - catalpol , aucubin , chemistry , rehmannia glutinosa , chromatography , iridoid , electrospray ionization , triple quadrupole mass spectrometer , ammonium formate , formic acid , selected reaction monitoring , tandem mass spectrometry , protein precipitation , glycoside , high performance liquid chromatography , mass spectrometry , stereochemistry , medicine , alternative medicine , traditional chinese medicine , pathology
The purpose of this study was to develop a method for simultaneous analysis of aucubin, catalpol, and geniposide, which are representative iridoid glycoside constituents of Rehmannia glutinosa , in rat plasma, urine, and feces using hydrophilic interaction ultra high‐performance liquid chromatography with tandem mass spectrometry. The three components were separated using 10 mmol/L aqueous ammonium formate containing 0.01% (v/v) formic acid and acetonitrile as a mobile phase by gradient elution at a flow rate of 0.2 mL/min, equipped with a Kinetex ® HILIC column (50 × 2.1 mm, 2.6 μm). Quantitation of this analysis was performed on a triple quadrupole mass spectrometer employing electrospray ionization and operated in multiple reaction monitoring mode. The chromatograms showed high resolution, sensitivity, and selectivity with no interference with plasma constituents. In all three iridoid glycosides, both the intra‐ and interbatch precisions (coefficient of variation %) were less than 4.81%. The accuracy was 96.56–103.55% for aucubin, 95.23–106.21% for catalpol, and 94.50–104.16% for geniposide. The developed analytical method satisfied the criteria of international guidance and was successfully applied to pharmacokinetic studies including oral bioavailability of aucubin, catalpol, and geniposide, and their urinary and fecal excretion ratios after oral or intravenous administration to rats. The new method was also applied to measure plasma protein binding ratios in vitro.