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Determination of gymnemagenin in rat plasma using high‐performance liquid chromatography–tandem mass spectrometry: application to pharmacokinetics after oral administration of Gymnema sylvestre extract
Author(s) -
Kamble Bhagyashree,
Gupta Ankur,
Patil Dada,
Khatal Laxman,
Janrao Shirish,
Moothedath Ismail,
Duraiswamy Basavan
Publication year - 2013
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.2845
Subject(s) - chemistry , chromatography , selected reaction monitoring , formic acid , extraction (chemistry) , pharmacokinetics , tandem mass spectrometry , mass spectrometry , elution , liquid chromatography–mass spectrometry , medicine
ABSTRACT A sensitive and rapid high‐performance liquid chromatography–tandem mass spectrometry (HPLC‐MS/MS) method has been developed and validated for the determination of gymnemagenin (GMG), a triterpene sapogenin from Gymnema sylvestre , in rat plasma using withaferin A as the internal standard (IS). Plasma samples were simply extracted using liquid–liquid extraction with tetra‐butyl methyl ether. Chromatographic separation was performed on Luna C 18 column using gradient elution of water and methanol (with 0.1% formic acid and 0.3% ammonia) at a flow rate of 0.8 mL/min. GMG and IS were eluted at 4.64 and 4.36 min, ionized in negative and positive mode, respectively, and quantitatively estimated using multiple reaction monitoring (MRM) mode. Two MRM transitions were selected at m / z 505.70 → 455.5 and m / z 471.50 → 281.3 for GMG and IS, respectively. The assay was linear over the concentration range of 5.280–300.920 ng/mL. The mean plasma extraction recoveries for GMG and IS were found to be 80.92 ± 8.70 and 55.63 ± 0.76%, respectively. The method was successfully applied for the determination of pharmacokinetic parameters of GMG after oral administration of G . sylvestre extract. Copyright © 2012 John Wiley & Sons, Ltd.