Analytical ApproachesIn the spring of 2010 BBSRC capital funding provided for the purchase of an AB Sciex 4000QTRAP mass spectrometer hyphenated with a Shimadzu Prominence HPLC system. This has been set up for LC/MS/MS analysis of sphingosine-1-phosphate from complex biomedical extracts, cholesterol and cholesterol ester analysis, direct infusion analysis of major lipids and the analysis of bis- and tris- phosphorylated phosphoinositides. This equipment has already been used in the analysis of several thousand samples.
A benefit of the high reliability of the whole system is its extensive use for the analysis of trace endogenous chemicals with biological significance. At the same time a Thermo TSQ Quantum Ultra GC-MS/MS was purchased and provides additional capability for example in investigating sterol metabolism.
Additional BBSRC capital funding in 2012 has facilitated the purchase of an Orbitrap Elite mass spectrometer which has replaced the Shimadzu IT-TOF. The Orbitrap is hyphenated with both a Shimadzu Prominence HPLC system and an Advion TriVersa NanoMate.
Theoretically, this equipment will enable the analysis of any biological chemical based on:
(a) our fully established HPLC separation methods which can separate different classes of neutral lipids, phospholipids and sphingolipids in a single run.
(b) the unique performance of Orbitrap Elite mass spectrometers which can carry out up to ten stages of tandem MS fragmentation and high resolution/accurate mass analysis at the same time.
Consequently, we expect that the resource will have the potential to identify novel classes of lipids/metabolites with important biological functions. To date the facility has operated primarily in a supporting role, however this enhanced capability permits the undertaking of novel discovery programmes.