期刊
ANALYTICAL BIOCHEMISTRY
卷 419, 期 2, 页码 323-332出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2011.08.045
关键词
Liquid chromatography-mass spectrometry; Multiswitching monitoring mode; Solid-phase extraction; Triterpenoidal saponins; Pulsatilla chinensis; Pharmacokinetics
资金
- Natural Science Foundation of Hebei Province-CSPC Pharmaceutical, China [C2011206167]
A novel method using high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) has been developed for the quantification of four triterpenoidal saponins (anemoside 84, pulsatilloside B, anemoside A3, and 23-hydroxybetulinic acid) in rat plasma following solid-phase extraction (SPE). The optimized procedure utilized off-line extraction of the analytes from plasma using polymeric (Strata-X) SPE cartridges. Detection and quantitation were performed by MS/MS using electrospray ionization (ESI) and multiple reaction monitoring (MRM) in a novel multiswitching monitoring mode. The analytes and internal standard (scutellarin) were analyzed using a Sapphire C18 column (250 x 4.6 mm, 5 mu m) with a linear gradient elution. The mass transition ion pairs of the triterpenoidal saponins were executed as follows: m/z 1219.7/749.4 for anemoside B4, m/z 819.4/347.2 for pulsatilloside B, m/z 749.6/471.2 for anemoside A3, m/z 471.4/471.4 for 23-hydroxybetulinic acid, and m/z 461.1/285.0 for the internal standard. The specificity, linearity, accuracy, precision, recovery, matrix effect, and stabilities were validated for all analytes in the plasma samples. In conclusion, the validation results demonstrate that this method is robust and specific. This validated method is a novel technique for sample preparation and quantitation and was successfully applied to estimate the pharmacokinetics of triterpenoidal saponins. (C) 2011 Elsevier Inc. All rights reserved.
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