Targeted lipidomics on HT-29 cells that were treated with and without propionate

Targeted lipidomics was performed on HT-29 cells that were treated with and without propionate. The goal was to assess how lipids were impacted with priopionate treatment. The experimental design included treating HT-29 cells (approximately 214800/well) with and without 0.2 mM propionate for 48 hours. The treatment media was EBSS, 5% FBS and p/s. There were four biological replicates for the control and treatment cohorts. Total lipid extracts from cell pellets were prepared using a modified MTBE lipid extraction protocol. Total lipid extracts were analyzed by liquid chromatography coupled to targeted tandem mass spectrometry (LC-MS/MS). The LC-MS/MS analyses were performed on an Ultimate 3000 Ultra High-Performance Liquid Chromatograph coupled to a Thermo TSQ Altis Tandem Quadrupole Mass Spectrometer (Thermo Scientific, San Jose, CA). LC-MS/MS methodology was adapted from the literature. LC-MS/MS data was acquired using Thermo’s Xcalibur software and data processing was achieved using Xcalibur 4.2 and TraceFinder 5.1.