Validation of UHPLC-MS/MS methods for the determination of kaempferol and its metabolite 4-hydroxyphenyl acetic acid, and application to in vitro blood-brain barrier and intestinal drug permeability studies

Fahimeh Moradi-Afrapoli, Mouhssin Oufir, Fruzsina R. Walter, M. Deli, Martin Smiesko, Volha Zabela, Veronika Butterweck, Matthias Hamburger

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Sedative and anxiolytic-like properties of flavonoids such as kaempferol and quercetin, and of some of their intestinal metabolites, have been demonstrated in pharmacological studies. However, routes of administration were shown to be critical for observing in vivo activity. Therefore, the ability to cross intestinal and blood-brain barriers was assessed in cell-based models for kaempferol (KMF), and for the major intestinal metabolite of KMF, 4-hydroxyphenylacetic acid (4-HPAA). Intestinal transport studies were performed with Caco-2 cells, and blood-brain barrier transport studies with an immortalized monoculture human model and a primary triple-co-culture rat model. UHPLC-MS/MS methods for KMF and 4-HPAA in Ringer-HEPES buffer and in Hank's balanced salt solution were validated according to industry guidelines. For all methods, calibration curves were fitted by least-squares quadratic regression with 1/X2 as weighing factor, and mean coefficients of determination (R2) were >0.99. Data obtained with all barrier models showed high intestinal and blood-brain barrier permeation of KMF, and no permeability of 4-HPAA, when compared to barrier integrity markers.

Original languageEnglish
Pages (from-to)264-274
Number of pages11
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume128
DOIs
Publication statusPublished - Sep 5 2016

Keywords

  • 4-HPAA
  • Blood-brain barrier
  • Caco-2 model
  • Kaempferol
  • LC-MS/MS
  • Method validation

ASJC Scopus subject areas

  • Analytical Chemistry
  • Drug Discovery
  • Pharmaceutical Science
  • Spectroscopy
  • Clinical Biochemistry

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