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Development of an LC–MS/MS method for quantification of two pairs of isomeric flavonoid glycosides and other ones in rat plasma: Application to pharmacokinetic studies
Author(s) -
Zhang Sixi,
Xie Yang,
Wang Jing,
Geng Yanmei,
Zhou Yu,
Sun Chengxin,
Wang Guangshu
Publication year - 2017
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.3972
Subject(s) - chemistry , chromatography , orientin , isoorientin , vitexin , formic acid , electrospray ionization , protein precipitation , mass spectrometry , tandem mass spectrometry , selected reaction monitoring , electrospray , acetonitrile , isovitexin , flavonoid , biochemistry , antioxidant
Abstract A liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated for simultaneous determination of six flavonoid glycosides – isoorientin ( 1 ), orientin ( 2 ), 2″‐ O‐β ‐ d ‐xylopyranosyl isoorientin ( 3 ), 2″‐ O‐β ‐ d ‐xylopyranosyl isovitexin ( 4 ), 6‐C‐ l ‐ α ‐arabipyranosyl vitexin ( 5 ) and vitexin ( 6 ) – in rat plasma using isoquercitrin as the internal standard (IS). Plasma samples were prepared by a one‐step protein precipitation with acetonitrile. Chromatographic analysis was carried out on a 25 cm C 18 column with a gradient mobile phase consisting of acetonitrile and 0.1% aqueous formic acid. Six analytes and IS were detected through electrospray ionization in negative‐ion selection reaction monitoring mode. The mass transitions were as follows: m/z 447.2 → 327.0 for 1 , m/z 447.2 → 327.0 for 2 , m/z 579.3 → 458.9 for 3 , m/z 563.0 → 293.1 for 4 , m/z 563.0 → 353.0 for 5 , m/z 431.1 → 311.1 for 6 , and m/z 463.1 → 300.2 for IS. Calibration curves exhibited good linearity ( r 2 > 0.9908) over a wide concentration range for all compounds. Intra‐ and inter‐day precision (RSD, %) at four different levels were both <14.2% and the accuracy (RE, %) ranged from −11.9 to 12.0%. The extraction recoveries of the six components ranged from 88.2 to 103.6%. The validated assay was successfully applied to the pharmacokinetic studies of the six components in male rat plasma after intravenous administration of total flavonoids of Scorzonera austriaca Wild.