Funding for a QTrap Mass spectrometer for the Cardiff Lipidomic Group.

Altered lipid metabolism is a common feature of most human diseases, including cardiovascular, cancer, inflammation, dementia and autoimmunity. Thus, accurate quantitation and identification of lipids in tissues and cells can provide significant insight into underlying mechanisms of pathology. Examples of lipid categories that are important for human disease include oxylipins, which comprise structurally diverse eicosanoids and prostaglandins generated by enzymes during inflammation, as well as sphingolipids and ceramides, lipids which are increasingly recognized to play key roles in cancer and organ and tissue development. Lipids make up a third of all known metabolites, and their sensitive and specific measurement is a major area of growing research need across the world, both for biomedical research but also many industrial applications. As evidence of this, the Wellcome Trust-funded resource LIPID MAPS (which we host at Cardiff) supports a global user base of around 65K researchers.

Accurate and sensitive quantitative analysis of lipids requires a biophysical method called mass spectrometry. This approach measures molecules based on their molecular weight and charge. This is most often preceded by chromatography, a method which first separates the lipids based on their physical properties in advance of their mass detection. Mass spectrometry of lipids is termed lipidomics, and with the growing appreciation of the importance of lipids to human disease, there is increasing need for provision of lipidomics across the UK and global research community. Due to its specialist nature, lipidomics MS is generally provided through access to facilities which have invested significant time and effort in both procuring equipment as well as the high level of technical support required. At Cardiff, O'Donnell has around 30 yrs experience in lipid biochemistry and has used MS for lipid structural studies since 1997, while Tyrrell has been based in the O'Donnell group for >13 yrs, including working as Facilities Manager. Our facility has been operational since 2005, hosting three instruments which analyse samples around the clock. However as the ageing QTrap is now being decommissioned, we risk losing a third of our total analytical capacity. This will critically impact our ability to support both our own research and that of our collaborators.

The Cardiff facility supports a combination of local research in our own group, other Cardiff-based researchers, and external work being conducted both in the UK and internationally. Below, we list our current collaborators, evidencing the breadth of our research and its relevance to MRC remit. Our instruments are available to all who have need for lipidomics, and we operate both cost-recovery and collaborative models depending on users' needs. Replacing the ageing 4000QTrap with the 7500QTrap would provide enormous benefits for our own research capacity, but importantly, our ability to support collaborative research internally and externally, as outlined in our case for support. Interest in lipidomics has exploded in recent years, and the demand on our expertise has grown exponentially. This new instrument will allow us to not only advance our own research but also give us the capacity to support more UK and international collaborations with the newest and highest throughput assays available.

The instrument requested is the 7500 QTRAP, which is Sciex's newest triple quadrupole MS, an instrument which allows absolute quantitation of small molecules to an extremely high level of sensitivity (fmol). Aside from reaching end-of-life, the older instrument does not have sufficient speed, pressure capability or sensitivity required for the new and continually evolving high throughput assays we require for our research. Several routine assays also depend exclusively on the old instrument and once it goes out of service, these would not be available anymore. The newer QTRAP functionality allows us to pair conventional MRM workflows with Enhanced Product Ion scanning allowing an extra level of confidence in acquired data. The speed and high sensitivity of this instrument with pos/neg mode switching and scheduling capability supports cutting edge assays that can measure up to 300 lipids in a single run, maximizing the data that can be acquired from a single sample and reducing the amount required, as well as dramatically increasing throughput through reducing the time of individual runs. This is ever important when measuring bioactive lipids that can often be present at very low amounts.

Lipidomics research at Cardiff is focused on understanding how lipids made by circulating blood cells drive thrombosis and vascular inflammation. Our local collaborators address research questions that encompass infection, immunity/inflammation, dementia, vascular disease, cancer and psoriasis. Our external (UK and international) collaborations study cardiovascular disease, sarcopenia, macrophage immunology, gut inflammation, cancer, sterol immunology, infection and lipid signaling. The Cardiff facility are members of external consortia which involve both provision of lipidomics MS analysis and training to the national and international lipidomics community. Full details of all are provided in our case for support.