Experimental platform demonstrates rapid fractionation and screening of Taxus brevifolia extracts, highlighting enhanced detection of bioactive anticancer natural products.
Key Points
To establish and validate an automated, high-throughput methodology for the production, analysis, and screening of natural product fraction libraries to accelerate drug discovery.
Integrated automated flash chromatography, solid-phase extraction, filtration, and parallel four-channel preparative HPLC to generate libraries of one to five compounds per well in 96- or 384-well plates.
Profiled fractions pre-screening using an eight-channel LC-ELSD-MS system and analyzed purified hits post-screening with mass spectrometry and microcoil probe NMR spectroscopy on 50 µg samples.
Evaluated library performance using Taxus brevifolia stem bark extracts screened against the National Cancer Institute's in vitro three-cancer-cell-line panel.
Uncovered active anticancer natural products from Taxus brevifolia fractions that were undetectable in the original flash chromatography fractions.
Successfully gathered structural NMR data from as little as 50 µg of active natural product fractions using a microcoil probe.