Profiling Intact Glycosphingolipids with Automated Structural Annotation and Quantitation from Human Samples with Nanoflow Liquid Chromatography Mass Spectrometry.
Ryan L SchindlerArmin OloumiJennyfer TenaMichael Russelle S AlvarezYiyun LiuSheryl GrijaldoMariana BarbozaLee-Way JinAngela M ZivkovicCarlito B LebrillaPublished in: Analytical chemistry (2024)
Sphingolipids are an essential subset of bioactive lipids found in most eukaryotic cells that contribute to membrane biophysical properties and are involved in cellular differentiation, recognition, and mediating interactions. The described nanoHPLC-ESI-Q/ToF methodology utilizes known biosynthetic pathways, accurate mass detection, optimized collision-induced disassociation, and a robust nanoflow chromatographic separation for the analysis of intact sphingolipids found in human tissue, cells, and serum. The methodology was developed and validated with an emphasis on addressing the common issues experienced in profiling these amphipathic lipids, which are part of the glycocalyx and lipidome. The high sensitivity obtained using nanorange flow rates with robust chromatographic reproducibility over a wide range of concentrations and injection volumes results in confident identifications for profiling these low-abundant biomolecules.
Keyphrases
- mass spectrometry
- liquid chromatography
- simultaneous determination
- induced apoptosis
- ms ms
- endothelial cells
- liquid chromatography tandem mass spectrometry
- tandem mass spectrometry
- high resolution mass spectrometry
- cell cycle arrest
- single cell
- high resolution
- high performance liquid chromatography
- high glucose
- gas chromatography
- induced pluripotent stem cells
- high throughput
- capillary electrophoresis
- pluripotent stem cells
- cell death
- solid phase extraction
- machine learning
- fatty acid
- quantum dots
- label free
- ultrasound guided