LipidCreator workbench to probe the lipidomic landscape - Platelet isolation and stimulation, Targeted Phospho and Glycerolipid Profiling

ABSTRACT:

Here we present LipidCreator, a software that fully supports targeted lipidomics assay development. LipidCreator offers a comprehensive framework to compute MS/MS fragment masses for over 60 lipid classes. LipidCreator provides all functionalities needed to define fragments, manage stable isotope labeling, optimize collision energy and generate in silico spectral libraries. We validate LipidCreator assays computationally and analytically and prove that it is capable to generate large targeted experiments to analyze blood and to dissect lipid-signaling pathways such as in human platelets.

In MTBLS1369, to validate the performance and accuracy of the transition lists provided by LipidCreator and their applicabability on other MS platforms, we used the transition lists for targeted profiling of phospho- and glycerolipids in human platelet material. Platelets were stimulated with 0.01 U/ml thrombin, 1 U/ml thrombin, 1 µg/ml CRP or 5 µg/ml CRP for 5 min. The MS profiling of unstimulated and stimulated platelet material was performed after HPLC separation on a Q-Exactive HF (Thermo Scientific, Bremen, Germany) in positive and negative mode, based on scheduled PRM. Samples were measured in triplicates, interspersed with blank / buffer and standard runs.

Studies linked to this manuscript include;

MTBLS1333; LipidCreator: Collision Energy Optimization and Fragment Intensity Prediction for Lipid Mediators - Thermo QExactive HF

MTBLS1334; LipidCreator: Collision Energy Optimization and Fragment Intensity Prediction for Lipid Mediators - Agilent QTof

MTBLS1369; LipidCreator: Platelet isolation and stimulation - Targeted Phospho- and Glycerolipid Profiling

MTBLS1375; LipidCreator: Targeted lipidomics analysis of Human Plasma samples and comparison with NIST SRM 1950 standard

MTBLS1376; LipidCreator: Targeted lipidomics analysis of S. cerevisiae to determine true and false identification

MTBLS1381; LipidCreator: Platelet isolation and stimulation - Targeted Lipid Mediator Profiling

MTBLS1382; LipidCreator: Platelet isolation and stimulation - DIA Lipid Mediator Validation

INSTRUMENT(S): Liquid Chromatography MS - negative - reverse phase, Liquid Chromatography MS - positive - reverse phase

SUBMITTER: Nils Hoffmann 

PROVIDER: MTBLS1369 | MetaboLights | 2020-04-15

REPOSITORIES: MetaboLights