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Quantitation of bergenin in human plasma by liquid chromatography/tandem mass spectrometry

Authors
Journal
Journal of Chromatography B
1570-0232
Publisher
Elsevier
Publication Date
Volume
877
Identifiers
DOI: 10.1016/j.jchromb.2008.11.005
Keywords
  • Bergenin
  • Lc–Ms/Ms
  • Pharmacokinetics
Disciplines
  • Pharmacology

Abstract

Abstract This paper reports the development and validation of an assay for quantitation of bergenin in human plasma using liquid chromatography/tandem mass spectrometry (LC–MS/MS). Bergenin and the internal standard (I.S.), 5-bromo-2,4(1 H,3 H)-pyrimidinedione (5-BrU), were separated by reversed phase HPLC and quantitated by MS/MS using electrospray ionization (ESI) and multiple reaction monitoring (MRM) in the negative ion mode. The most intense [M−H] − MRM transition of bergenin at m/ z 326.9 → 312.3 was used for quantitation and the transition at m/ z 188.9 → 42.2 was used to monitor 5-BrU. Stability issues with bergenin required the addition of ascorbic acid to plasma samples prior to storage and analysis within 10 days storage at −80 °C. The method was linear in the range 3–1000 ng/mL with intra- and inter-day precision of 3.94–5.96 and 1.62–8.31%, respectively, and accuracy <2.33%. The assay was successfully applied to a pharmacokinetic study in healthy volunteers after administration of a single 250 mg oral dose.

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