A high sensitivity triple quadruple mass spectrometer coupled to an ultra-high pressure liquid chromatograph system for quantitative analysis

Lead Research Organisation: University of Warwick
Department Name: Chemistry

Abstract

The triple quadrupole mass spectrometer based LC-MS/MS system has been widely adopted as the most sensitive methodology for the quantitation of small and large organic molecules in complex biological samples for both industrial and academic research laboratories. The triple quadruple MS system has other unique capabilities, including precursor ion scan and neutral loss scan with high sensitivity, which are important tools for disease biomarker and bioactive natural products discoveries. Compounds that have a similar structure, either with identical backbone or functional groups, can be identified using the targeted scan option offered from a triple quadruple mass spectrometer. Despite the fact that Warwick Chemistry has one of the largest mass spectrometry facilities in the UK, consisting of multiple ESI-Q-TOF, MALDI-TOF and ESI-Ion trap mass spectrometers, we lack an instrument that is suitable for ultra-high sensitivity quantitative analysis, and this is hindering us from acquiring important information from biological samples. We urgently require a triple quadruple type of mass spectrometer for LC-MS/MS based quantitation in multiple reaction monitoring mode.
The new instrument will enable us to significantly expand and enhance our current research activities in many RCUK (mainly BBSRC) funded projects. This will include metabolomics studies on British ash trees (BB/N021452/1, Led by Prof Murray Grant), where the aim is to discover reliable biomarkers for ash dieback disease, and establish a database for British ash tree metabolome. In turn this will pave the way for future disease diagnosis and the selective cultivation of a pathogen resistant genome type ash tree. Another major BBSRC funded project (sLoLa BB/K002341/1, led by Prof Gregory Challis) aims to discover bioactive agrichemicals from bacteria through microbial genome mining. The unique precursor ion scan and neutral loss scan capabilities on the proposed instrument will enable swift identification of compounds that belong to the same family, or which have the same functional groups, with high sensitivity. Many other projects including the quantitation of novel organometallic anticancer drugs, anti-inflammatory drugs and anti-viral drugs, as well as the investigation of their metabolism, will also benefit from accessing the proposed instrument.
The new instrument will also enable us to train the next generation of scientists on the most advanced mass spectrometer, which will enable them to gain hands-on experience that will be extremely beneficial for their future career development. Our research led teaching means that even undergraduate students can also benefit from the access to a state of the art facility.
The proposed instrument will be incorporated into, and will complement and significantly enhance our existing mass spectrometry facility though the addition of new capabilities. The new MS will not only benefit our students and staff in Warwick, our academic and industrial collaborators will also benefit from receiving training, and access to the most advanced MS technologies.

Technical Summary

Since the commercial introduction of soft ionisation techniques, particularly electrospray ionisation (ESI) and matrix assisted laser desorption ionisation (MALDI) (for which Fenn and Tanaka shared half of the 2002 Nobel Prize in Chemistry), mass spectrometry has truly become one of the most important research tools not only for chemists, but also for a wide range of other research fields. Due to its non-scanning nature, triple quadruple mass spectrometer based LC-MS/MS quantitation in multiple reaction monitoring mode is arguably the most sensitive way to quantify many different types of compounds. The proposed triple quadruple mass spectrometer based LC-MS/MS system will complement our existing MS facility, providing new capability in both quantitative and qualitative analysis, and significantly enhance our research outcomes in multiple BBSRC funded projects.
Through a competitive tendering process, we will evaluate and select the most suitable instrument for our applications, including BBSRC funded projects on bioactive natural product discovery (SLoLa BB/K002341/1; BB/P002560/1) and the discovery and quantification of disease biomarker for ash die-back (BB/N021452/1). The new instrument will be incorporated into our existing mass spectrometry facility in the Department of Chemistry. Once it is fully operational, current MS facility staff will receive advanced training from the vendor on method development, operation and maintenance of the new instrument. Full training will be provided to all future users by MS facility staff. Instrument access will be managed using a on-line booking system, and trained users can book and use the instrument 24/7. Project prioritisation and future research development will be managed by Lijiang Song and overseen by the steering committee consisting of all the applicants. Through Warwick Scientific Services and existing academic and industrial collaborations, external users can receive training and access to the new facility.

Planned Impact

As one of the UK's leading research-intensive Universities, Warwick has an excellent equipment base to support scientific research. However, with technologies advancing so rapidly today, it is important that we constantly upgrade and enhance our research facilities to ensure that we remain at the cutting edge of technologies and can provide students and staff members with optimal training and research experience. The new mass spectrometer requested in this application will be incorporated into our existing Chemistry Department mass spectrometry facility, enabling enhanced capacity and capability in key areas where we currently experience high demand from research groups across departments within Warwick, and from industrial and academic collaborators across the UK and beyond. The University of Warwick is committed to shared infrastructure/facilities with ready access for trained researchers, with core support provided from the University, from the academic departments and from research grant and contracts income. The new instrument will be managed by an expert custodian with extensive technical expertise and facility management experience to ensure that the equipment is properly maintained and operated, and that usage is maximized through a highly efficient on-line booking system.
Significant economic impacts across many of the BBSRC priority themes are likely, including agriculture and food security, industrial biotechnology and bioenergy, biosciences for health. We interact strongly with UK industry by way of collaborative research projects and contract analytical work through Warwick Scientific Service (WSS).
To train the next generation of research scientists for academia and industry, we require access to state of the art facilities. Since 2006, Warwick Chemistry has adopted open access policy towards many of our research facilities. We provide comprehensive training to all users on advanced analytical equipment. Through the operation of advanced equipment, our students and researchers are not only gaining a deep understanding of the technology and hands-on experience, but more importantly they gain a better understanding of their research field, which makes Warwick graduates some of the most employable in the UK. Consequently, many of our postgraduates and PDRAs enter employment in the industry sector after their studies. With an addition of a triple quadrupole mass spectrometer which is widely used by industrial research laboratories, it will mean that our students and staff can be even better equipped with the knowledge and experience to take on future research challenges.
Research-led teaching has been a popular way in delivering course content in Warwick. It enables students to apply their newly acquired text book knowledge to real world problems by getting involved in cutting edge research. By working in the laboratory with postgraduate/postdoctoral researchers, undergraduates can also benefit from using the most advanced quantitative analysis facility, and gain a deeper understanding of the theoretical knowledge they learn in the classroom.
Warwick Chemistry has a dedicated outreach officer (Senior teacher fellow, Mr Nick Barker) who carries out a wide range of activities with school age children, such as delivering lectures in schools and organising vists of school pupils to the department. We will use this expert resource to explain the aims and outcomes of this research to wider society, especially school children through our schools outreach programme.

Publications

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Description Been a equipment grant, we have the proposed instrument delivered and fully installed on time after an extensive and comprehensive evaluation process. The instrument is fully operational now, wide range of users will be trained on the instrument in the coming month, many projects, including those funded by the BBSRC will begin to benefit from the access to this high sensitivity triple quad based LC-MS/MS system to carry out both quantitative and qualitative analysis.

Since the last report, we have had advanced user training to 3 members of the MS facility staff by the vendor Waters. We have also trained users from both the department of Chemistry and the School of Life Sciences, the total number of users trained on this instrument is currently14 and will increase over time. We have carried out lots of method development for different projects/applications in the last year, including one project headed by Prof Liz Wellington from SLS (the School of Life Sciences) on antimicrobial degradation in the environment; two projects headed by Prof Murray Grant and myself (SLS-Chemistry) on potential biomarker discovery and quantitation for a BBSRC funded project on Ash Dieback; bioactive natural product discovery from a soil fungal strain Trichoderma hamatu also funded by the BBSRC; A project headed by myself to work on bioactive natural product discovery from marine microorganism funded by the Royal Society; two small projects funded by industry (Unilever and Lytegro ) have also made use of this facility to quantify small molecules in complex samples. I have also started a new collaboration with Prof Murray Grant (SLS) and Prof Pradeep Kachroo from the University of Kentucky to work on the important signaling molecule pipecolic acid in plant samples, we make use of this new instrument to reliably quantify the target compounds.
Exploitation Route At the earlier stage of this facility been installed, PDRAs and PhD students on BBSRC funded projects will be trained on the new instrument and start to acquire data for their research. In the next few months, we will also try to make this available to other researchers in the department/University, by either providing instrument time to them or to provide training to them, so more can benefit from access to this newly added capacity.

After one year of operation, we have 14 trained users including student, PDRA and staff as well 6 research projects (mostly funded by UKRI or industry) started to make use of this very valuable tool. We anticipate more users will be trained in the future once we have demonstrated the usefulness of this unique instrument and make people aware of this new capability/capacity through conference/meeting presentation and publication. Therefore, the new instrument will not only significantly enhance our existing research, it will also bring new collaborations and broaden our research field.
Sectors Agriculture, Food and Drink,Chemicals,Education,Environment,Healthcare,Pharmaceuticals and Medical Biotechnology