Planta Med 2012; 78 - PJ81
DOI: 10.1055/s-0032-1321241

Combination of bioautography with HPTLC-MS/NMR: A fast identification of AChE inhibitors from Galbanum

HR Adhami 1, U Scherer 2, H Kaehlig 3, T Hettich 2, G Schlotterbeck 2, E Reich 4, L Krenn 1
  • 1Department of Pharmacognosy, University of Vienna, Vienna, Austria
  • 2Institute for Chemistry and Bioanalytics, University of Applied Sciences Northwestern Switzerland, Muttenz, Switzerland
  • 3Institute of Organic Chemistry, University of Vienna, Vienna, Austria
  • 4CAMAG Co. Laboratory, Muttenz, Switzerland

Isolation of bio-active compounds from plant extracts is laborious and time-consuming. A dichloromethane extract of galbanum, the oleo gum-resin from Ferula gummosa Boiss., had shown AChE inhibitory activity (1). For fast identification of active compounds, HPTLC bioautography was combined with HPTLC-MS/NMR. After pre-fractionation of the extract by vacuum liquid chromatography, fractions were separated by automated HPTLC and active zones determined by bioautography. The mobile phase was individually optimized for each active fraction according to the CAMAG guideline and automatic detection of the compounds by UV absorbance was performed in a TLC scanner. A CAMAG TLC-MS interface was used to elute the single compounds from the plates directly into the mass spectrometer. In parallel, the most concentrated active zones were extracted from HPTLC plates for one and two-dimensional 1H and 13C NMR and QTOF-MS. The most abundant compound with AChE inhibitory effects was identified as auraptene. The structure elucidation of other compounds is in progress. This is the first report of auraptene in Ferula gummosa. HPLC analyses carried out to determine the concentration of auraptene in the extract.

Reference: 1. Adhami HR, Farsam H, Krenn L. 2011. Phytother Res. 25: 1148–1152.