Red Blood Cell Lipidomics analysis through HPLC-ESI-qTOF: application to red blood cell storage
Recent developments in mass spectrometry (MS) have enabled fast and sensitive detection of lipid species in different biological matrices.
In the present study we performed an on-line HPLC-microTOF-Q MS approach to the red blood cell (RBC) lipidome. We thus exploited bioinformatic tools for the interrogation of novel databases, such as LIPID MAPS. By means of ad hoc software suites for mass spectrometry-based metabolomics analyses, we could address the key biological issue of the RBC lipidome, within the framework of RBC storage for transfusion purposes. Samples were collected from subjects living in the province of Viterbo, where olive oil consumption represents a central aspect of the diet. On this ground, we could postulate a diet specific effect on the accumulation of lipid-specific storage lesions.
The analyses yielded the tentative identification of a huge number of lipid molecules on the basis of accurate intact mass values and retention times, and MS/MS validation. This analytical workflow was exploited to consolidate existing knowledge on the RBC lipid composition and individuate statistically significant fluctuations of lipids throughout storage duration of RBC concentrates under blood bank conditions. Our analysis indicated ceramides, glycerophospholipids and sterols as key targets of RBC storage lesions to the lipidome, that will deserve further targeted investigations in the future. It also emerged how compositional analyses of the RBC lipidome might end up yielding different results on the basis of the background of the blood donor (i.e. diet), which might translate into region-specific lipidomic alterations over storage progression of RBC concentrates.