Ultra-high-throughput mass spectrometry for quantitative metabolomics

Metabolomics refers to the quantitative measurement of the complement of small molecules (metabolites) found within a biological system. One way to identify such molecules relies on the determination of their masses, known as mass spectrometry. Mass spectrometry (MS), coupled to prior separation with either liquid or gas chromatography, dominates the metabolomics field for the analysis of these structurally diverse metabolites. However, this comes with a significant cost because the chromatography step, while valuable for separation of compounds, adds significant time to the analysis: 30 minutes per sample is typical.

Three years ago we established the Centre for Metabolomics Research (CMR) within the University of Liverpool. In the CMR we are developing and applying metabolomics in many different fields including (amongst others) human physiology and healthy ageing, antimicrobial resistance, synthetic biology, industrial biotechnology and for the enhanced understanding of mammalian systems and crop development and resilience. We are also training the next generation of interdisciplinary metabolomics scientists and we are proud of our capabilities here.

Our focus within the CMR has been to develop MS-based metabolomics, with up front 'slow' chromatography - this limits high sample throughput analysis as there is simply not enough time in the day to make these measurements. Therefore, what we still fundamentally lack are ultra-high-throughput methods for the analysis of specific metabolites.

The instrument requested is the Echo Mass Spectrometry System from SCIEX. This is vital for us to be able to enhance our metabolomics capabilities and will turbo charge the speed of our metabolomics work with applications in many areas of bioscience research.

We can confirm that this instrumentation is not available in CMR, the University of Liverpool, the North West in general, and indeed the manufacturers have confirmed that this will be the first such instrument embedded in any academic institution in the UK.

The aim of this proposal is thus to secure this Echo MS-based instrument for an extensive User Community in Liverpool and elsewhere with ultra-high-throughput capability. With support from the University of Liverpool Shared Research Facilities this platform would be made available to other institutes as well as industrial partners thus enhancing the wider UK science base.

We propose a unique, multi-purpose mass spectrometer with applications in many areas of bioscience research. The instrument is the ultra-high-throughput Echo MS from SCIEX that has been designed precisely for these purposes.

The Echo MS incorporates a fluidics module that allows sampling rates of up to 3 samples per second using acoustic droplet ejection for delivery of 2.5 nL of sample from 384 or 1536 microplates. Electrospray ionisation is achieved via the Optiflow Turbo C Ion Source and is integrated with the SCIEX Triple Quad 6500+ MS/MS for mass analysis. This system is the only commercially available integrated system with acoustic droplet ejection coupled to MS and the speed of the analysis will cause a step change in the sample throughput needed for targeted quantitative analysis within the metabolomics pipeline.

All applicants from University of Liverpool are well-funded (mainly BBSRC) and engaged in a variety of BBSRC-supported research programmes including: understanding bacterial adaptation in antimicrobial resistance; synthetic biology projects including fine chemical production, enhanced CO2 fixation, function and regulation of cyanobacterial bioenergetic machinery; metabolite transporters; inhibition of phosphorylation and sulfation pathways linked to cellular metabolism; crop improvement; as well as human health and pharmacometabolomics linked to genomics. The projects are described in more detail within the Case for Support.

Once the system is in place and optimised it will be highly beneficial to the University of Liverpool, the local community, and the North West in general. With infrastructure support from the University of Liverpool Shared Research Facilities (LIV-SRF), the proposed Echo MS from SCIEX will provide significant infrastructure and training for many research groups at Liverpool and will be made available to BBSRC-funded researchers from other academic institutes as well as industrial scientists.