High throughput screening assay for quantification of Cer d18:1/16:0, d18:1/24:0, d18:1/24:1, d18:1/18:0, d18:1/14:0, d18:1/20:0 and d18:1/22:0 ceramides in HepG-2 cells using RapidFire Mass spectrometry.

High throughput screening assay for quantification of Cer d18:1/16:0, d18:1/24:0, d18:1/24:1, d18:1/18:0, d18:1/14:0, d18:1/20:0 and d18:1/22:0 ceramides in HepG-2 cells using RapidFire Mass spectrometry. Biomed Chromatogr. 2019 Dec 28;:e4790 Authors: Dittakavi S, Mahadevan L, Chandrashekar DV, Bhamidipati RK, Suresh J, Dhakshinamoorthy S, Li Z, Baerenz F, Tennagels N, Mullangi R Abstract Ceramides are known to be involved in various biological processes and their physiological levels are elevated in various disease conditions like diabetes, Alzheimer's, atherosclerosis etc. To facilitate the rapid screening of Cer d18:1/16:0, d18:1/24:0, d18:1/24:1, d18:1/18:0, d18:1/14:0, d18:1/20:0 and d18:1/22:0 inhibition in HepG-2 cells, a RapidFire coupled to tandem mass spectrometry (RF-MS/MS) assay method has been developed. The RapidFire platform provides an automated solid-phased extraction system that gave a throughput of 12.6 sec per sample to an LC-MS/MS using electrospray ionization under positive-ion mode. Chromatographic separation of Cer d18:1/16:0, d18:1/24:0, d18:1/24:1, d18:1/18:0, d18:1/14:0, d18:1/20:0 and d18:1/22:0 was achieved using a ternary gradient on C8 type E cartridge. The MS/MS ion transitions monitored were 538.2→264.2, 650.7→264.2, 648.6→264.2, 566.4→264.2, 510.4→264.2, 594.4→264.2, 622.5→264.2 and 552.3→250.2 for Cer d18:1/16:0, d18:1/24:0, d18:1/24:1, d18:1/18:0, d18:1/14:0, d18:1/20:0 and d18:1/22...
Source: Biomedical Chromatography : BMC - Category: Biomedical Science Authors: Tags: Biomed Chromatogr Source Type: research