Receptor-based high-throughput screening and identification of estrogens in dietary supplements using bioaffinity liquid-chromatography ion mobility mass spectrometry.
作者： Payam Aqai , Natalia Gómez Blesa , Hilary Major , Mattia Pedotti , Luca Varani
DOI: 10.1007/S00216-013-7384-1
关键词: Electrospray 、 Chromatography 、 Tandem mass spectrometry 、 Chemistry 、 Ion source 、 Liquid chromatography–mass spectrometry 、 High-throughput screening 、 Mass spectrometry 、 Ligand binding assay 、 Ion-mobility spectrometry
摘要: A high-throughput bioaffinity liquid chromatography-mass spectrometry (BioMS) approach was developed and applied for the screening identification of recombinant human estrogen receptor α (ERα) ligands in dietary supplements. For screening, a semi-automated mass spectrometric ligand binding assay applying 13C2,15 N-tamoxifen as non-radioactive label fast ultra-high-performance–liquid chromatography–electrospray ionisation–triple-quadrupole-MS (UPLC-QqQ-MS), operated single reaction monitoring mode, readout system. Binding to ERα-coated paramagnetic microbeads inhibited by competing estrogens sample extract yielding decreased levels UPLC-QqQ-MS. The showed high ionisation efficiency positive electrospray (ESI) so BioMS is able screen supplements despite their poor both negative ESI modes. performed 96-well plate, all these wells could be measured within 3 h. Estrogens suspect extracts were identified full-scan accurate collision-cross section (CCS) values from UPLC-ion mobility-Q-time-of-flight-MS (UPLC-IM-Q-ToF-MS) equipped with novel atmospheric pressure source. Thanks ion source, this instrument provided picogram sensitivity mode an additional point (experimental CCS values) next retention time, tandem data. combination UPLC-QqQ-MS UPLC-IM-Q-ToF-MS provides extremely powerful analytical tool early warning ERα bioactive compounds demonstrated analysis selected which different identified.
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