In-situ clean-up and oplc fractionation of chamomile flower extract to search active components by bioautography

Abstract

Bioassay-guided isolation of antibacterial components of chamomile flower methanol extract was performed by overpressured layer chromatography (OPLC) with on-line detection, fractionation combined with sample clean-up in-situ in the adsorbent bed after off-line sample application. The antibacterial effect of the eluted fractions and of those compounds remaining on the adsorbent layer after separation was tested with direct bioautography (DB) against the bioluminescent Pseudomonas savastanoi pv. maculicola and Vibrio fischeri. The fractions with high biological activity were analyzed by solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) and liquid chromatography and tandem mass spectrometry (LC-MS/MS). Two active uneluted compounds were characterized by off-line OPLC-MS using a thin-layer chromatography (TLC)-MS interface. Mainly, essential oil components, coumarins, flavonoids, phenolic acids, and fatty acids were identified in the active fractions. © Akadémiai Kiado, Budapest.