[1] Phillips, M., et al., Point-of-care breath test for biomarkers of active pulmonary tuberculosis. Tuberculosis, 2012. 92(4): p. 314-20.

Prof. John Lindon

Bringing molecules into medicine using metabonomics: the challenges of patient phenotyping for diagnosis, optimising treatment and prognosis

John C. Lindon

Biomolecular Medicine, Department of Surgery and Cancer, Imperial College London, Sir Alexander Fleming Building, South Kensington, London SW7 2AZ UK

NMR- and MS-based based metabolic profiling (metabonomics) of mammalian biofluids and tissues has proved to be a useful approach for understanding physiological effects, for disease diagnosis, for prognosis of future treatment and outcome, and for the investigation of interactions between the host and its gut microbial ecology.  Such applications require the employment of multivariate statistics for efficient data interpretation. This talk will provide a brief overview of metabonomics technologies, provide some recent examples of clinical metabonomics, and highlight new developments in metabolic phenotyping of hospital patients with the aim of achieving stratified or more personalised treatment. This is a challenging task and if the approach is also to be used for real-time medical and surgical applications then new technologies and organisational approaches will be needed and these will be discussed. 

Prof. Jorg Baumbach

Volatile metabolites of humans, animals, cells and bacteria - detected using ion mobility spectrometry

Jörg Ingo Baumbach

B&S Analytik GmbH, BioMedical Center Dortmund, Otto-Hahn-Str. 15, 44227 Dortmund and Technical University Dortmund, Fakulty Bio- and Chemical Engineering, Emil-Figge-Str. 70, 44227 Dortmund

Volatile metabolites in human exhaled breath, in the exhale of animals or over cell cultures and bacteria could be detected down to the pg/L-range (pptv) using ion mobility spectrometry coupled to multi-capillary columns within less 10 minutes total analysis time and in rather complex und humid matrixes. Normally, about 10 mL of exhaled breath will be analysed and characterized with respect to identification and quantification. Some proven medical applications will be considered (e.g. lung cancer, anaesthesia, SEPSIS, bacterial infections) and recent results in metabolomics and breath gas analysis will be presented. The advantages and disadvantages compared to GC/MS are discussed with respect to information content, analysis time, instrumentation requirements and analytical and separation power. In addition, the improvement of results using modern methods of bioinformatics will be shown (including data mining, statistics, decision trees and correlation analysis).

Dr. Tony Bristow

When Big is Brilliant and Small is Beautiful – Mass Spectrometry diversity for a huge range of analytical challenges.

Tony Bristow¹, Andy Ray¹, Marie-Claire Lacassin¹, Hilary Major², Jackie Mosely³ Colin Creaser⁴ and Peter O’Connor⁵.

¹ Pharmaceutical Development, AstraZeneca, Macclesfield, SK10 2NA^{, 2} Waters Corporation, Wythenshawe, Manchester, M23 9LZ, ³ Department of Chemistry, University of Durham, Durham, DH1 3LE, ⁴ Department of Chemistry, Loughborough University, Loughborough, LE11 3TU, ⁵ Department of Chemistry, University of Warwick, Coventry, CV4 7AL.

The range of analysis problems to which mass spectrometry can be applied continues to grow. This expansion has resulted from the continued innovation in the development of all aspects of mass spectrometry instrumentation. This includes ionisation techniques, mass analysers and new experiments. This presentation will describe a number of recent mass spectrometry innovations that have been investigated for a diverse range of analytical challenges within the pharmaceutical industry. The examples will include the application of novel ion source designs, on-line mass spectrometry, advanced mass analysers, novel techniques for structural characterisation and new approaches for the characterization of complex polymeric systems. The impact of these developments will be discussed.

Prof. David Cowan

London 2012 - Olympic and Paralympic Games – the latest science behind the anti-doping tests

David Cowan

Department of Forensic Science & Drug Monitoring, Kings College, London, UK

London 2012 is now but a memory but one that will live with the world for many years.  What of the science?  Many advances were made in preparation for delivering a world-class anti-doping analytical service.  Our goal was to deliver a programme of such strength that it would help deter drug misuse.  Put simple, our task was encapsulated in the phrase “super-fast and super-sensitive”.  We developed an infrastructure that would allow us to analyse more than 6,000 samples in less than six weeks with turnaround times as short as 24 hours.  In order to achieve these goals we made of use of tandem MS, UHPLC, high resolution MS providing total data capture, sensitive combustion isotope ratio mass spectrometry through to biomarker testing for the administration of human growth hormone.  Most of the results are now in the public domain and a personal insight will be provided in this presentation.

Prof. Peter Fielden

Cheap as Chips? - Analysis with Low Cost Sensing Systems

Peter Fielden

Department of Chemistry, University of Lancaster, UK

Sensing systems based on polymer substrates may be fabricated at very low cost by manufacturing techniques, such as injection moulding,  normally associated with mass production. Whilst such systems cannot ultimately compete with high-end laboratory-based analytical techniques, there are many examples where sensing systems can provide sensitive and selective assays, with the added attraction of portability. Microseparations, such as chip-based isotachophoresis, coupled to simple sensors may be employed to address a significant range of target species within complex samples. Electrochemical assays have been demonstrated where plastic composite and screen-printed ink-based electrodes have been used, rather than expensive rare metals such as gold and platinum, to fabricate electroanalytical sensing systems. Optical microresonator sensors may be fabricated in a microreactor as spheres, or by stereolithography as rings, with specificity added through molecular imprinting or immunoassay chemistry. Consideration of the direction and future of low-cost sensing systems will be made.

Dr. Sandra Groscurth

Structure Elucidation and verification with different analytical methods- a case study

Sandra Groscurth

Bruker Switzerland

Mass Spectrometry (MS), Nuclear Magnetic Resonance (NMR) and Single Crystal X-ray Diffraction (SC-XRD) are known to be extremely powerful analytical methods to verify and elucidate molecular structures. Therefore they form essential tools for successful chemical and biological research.

Today’s hardware and software has made analytical data acquisition more convenient and efficient than ever before. However, subsequent experimental data analysis can be time consuming and is often the bottleneck in the structure verification and elucidation process. Therefore Bruker has focused on the development of software packages assisting the chemist with the interpretation of experimental data. Based on a case study, we present these developments in the process of computer-assisted structure verification and elucidation with different analytical techniques, including MS, NMR and X-ray crystallography.

Dr. G. John Langley

Unravelling complexity through experiment design

John Langley

Department of Chemistry, University of Southampton

Generic hyphenated methods have become common place and can solve a number of issues, e.g. high throughput needs, analysis of analogues etc. These approaches use generic methods and as such often ignore the needs of the specific analyte.  By considering the analytical need and the analyte(s) in question, the optimum hyphenated technique can be determined.  This presentation will discuss biodiesel analysis at Southampton, in particular oxidation of FAMEs, through the use of HPLC-MS, GC-MS and SFC-MS (and MS-MS) to aid structural elucidation.

Simon Noble

The challenges of high sensitivity MS detection in Bioanalysis

Simon Noble

Quotient Bioresearch Ltd.

 As drugs become more potent and the use of low abundance endogenous biomarkers becomes more prevalent in drug development, the development and validation of sensitive mass spectrometry (MS) based bioanalytical methods is increasingly key to providing the necessary pharmacokinetic and pharmacodynamic data.  This presentation will look at the challenges of high sensitivity MS methods, and how the structures of the analytes relate to the various strategies in extraction, chromatography and detection are employed to achieve them.

Dr. Madalina Oppermann

m/z Cloud - Novel Spectral Library for Metabolite Identification

Robert Mistrik¹; Juraj Lutisan¹; Yingying Huang²; Rose Herbold²; Madalina Oppermann³; Eric Genin⁴

¹HighChem, Ltd., Bratislava; Thermo Fisher Scientific, ²San Jose, CA, ³Stockholm and Paris⁴

 Analysis of numerous small molecules and structural assignments of individual metabolic components is a major bottleneck in various areas of metabolomics. In mass spectrometry widely used library search systems are designed to identify compounds represented in the reference library. If the unknown compound is not represented in the library, the compound cannot be identified by this method. We will present a new type of mass spectral library providing the functionality required for elucidation of unknowns even if compounds are not present in the library.

A few years ago, Precursor Ion Fingerprinting (PIF) was developed. This approach identifies sub-structural information through the comparison of product ion spectra of structurally related compounds. Structural information is derived by utilizing previously characterized ion structures stored in reference libraries of tandem mass spectral data and matching them with unknown product ion spectra. PIF is a powerful technique that takes advantage of the structural continuum and conservation of eukaryotic metabolism.

The library is implemented in a relational database that will be accessible through a public domain web site of an emerging consortium named “m/z Cloud”. Since the consortium is predominantly oriented towards high-resolution, accurate mass spectra, the database design, spectral management, and library search algorithms require a completely new architecture compared to traditional spectral databases. The idea of this project is to create a library of comprehensive spectral ion trees based on structurally characterized product ion spectra to enable the identification of substructures in unknowns.

Each structurally characterized product ion spectrum will contain the precursor ion m/z value, a list of product ion m/z values with mass accuracies, corresponding absolute and relative intensities, ion polarity, charge state, the structure of the precursor ion, and the structure of the parent molecule. For the assignment of fragment structures to a precursor ion in the process of creating structurally characterized product ion spectra, it is extremely beneficial to have high-resolution spectra since the accurate m/z values of precursor and product ions greatly reduce the number of possible molecular formulas for fragment structures. Also, the determination of the structural arrangement for the elucidated molecule benefits from exact mass measurements by constraining the elemental composition of the elucidated molecule and

consistently validating the calculated mass of recognized fragment structures and accurate m/z values of precursor and product ions.

The m/z Cloud public domain database aims to provide complete library technology based on spectral ion trees to enable elucidation of unknowns using the precursor ion fingerprinting method.

Dr. Andrew Phillips

The use of NMR for Trace Analysis within Pharmaceutical Development

Andrew R. Phillips, Ian C. Jones and Steve Coombes

Pharmaceutical Development, AstraZeneca, Macclesfield UK

NMR is a well-established quantitative technique for looking at relatively small amounts of material – for example residual solvents within an active pharmaceutical ingredient down to levels of 0.1% w/w. Recently there has been considerable concern from pharmaceutical regulatory agencies over the control of potential genotoxic impurities (PGI) in medicinal products.  The Threshold of Toxicological Concern (TTC) for PGIs in commercial products is 1.5 μg/day, or single-figure ppm with respect to a typical drug substance.  Consequently, methods for the measurement of impurities in the single-ppm range are required – presenting a significant analytical challenge. We have recently shown that NMR can detect down to these low level often with significant advantages over other more traditional techniques in terms of method development, sample preparation and experiment time. [1] The key to the success of NMR is overcoming the inherent lack of sensitivity so this presentation will focus on some of the important factors that determine the level of detection, such as the performance of the NMR system, substrate concentration, linewidth, resolution and dynamic range.  This will include how new state of the art equipment in our laboratory has been utilised to overcome some of these challenges. In addition a number of new genotoxic impurities examples analysing both pure compounds and formulated products will be presented.  Furthermore it will shown that the use of NMR can be expanded to other trace analysis problems such as cleaning validation: confirming vessels used for chemical reactions are not contaminated with material from previous experiments.

[1] Andrew R. Phillips ‘Analysis of Genotoxic Impurities by Nuclear Magnetic Resonance Spectroscopy’ in ‘Genotoxic Impurities: Strategies for Identification and Control edited by Andrew Teasdale, 2011, Wiley-Blackwell

Dr. Hiroaki Sasakawa

New Solid NMR Probes and Applications for Structure Analysis

Hiroaki Sasakawa

JEOL UK Ltd, JEOL house, Silver Court, Watchmead, WelwynGarden City, Herts, UK

We released a new 0.75 mm solid state NMR probe on last April. Its probe is capable of ¹H high resolution analysis by sample spinning at 110 kHz. This new probe is expected to be effective in applications that are difficult for conventional NMR systems, including micro analysis of drugs, natural products, and thin films. We will show some example data with this new probe. Also, we released 8 mm solid state NMR probe for narrow bore magnet. This is very large volume MAS probe (606 ul). Its sensitivity boosts by a factor of around 4, comparing to 3.2 mm MAS probe. This means more than 10 times shorter experiments and promises very quick observation of small peaks. In case of large volume MAS probe, it is not easy to adjust resolution to get high quality spectra, but we developed a simple method to automatically shim at the magic angle spinning [1]. This is introduced based on the gradient shimming approach. We can achieve the less than 1 Hz line width on ¹³C NMR adamantane within few minutes.

[1] Y. Nishiyama, Y. Tsutsumi, and H. Utsumi, J. Magn. Reson. 216 (2012) 197-200.

Prof. Aldrik Velders

Nanolitre NMR Spectroscopy

Aldrik Velders

BioNanoTechnology Group, Wageningen University & Research Centre , Wageningen, The Netherlands

 Analysis of mass-limited samples with NMR spectroscopy is a major challenge, which has triggered the development of expensive and technologically demanding solutions, such as cryoprobes, ultra-high magnetic fields and hyperpolarization strategies. A relatively cheap alternative approach regards the use of miniaturized coils and, over the past two decades, microcoils of different geometries have proven this to be a successful strategy.[1] We are particularly interested in microfluidic chip designs with planar spiral transceiver coils and detection volumes in the lower nanoliter range. These chips can be used in static mode, e.g. for observation of supramolecular interactions by 19F NMR spectroscopy,[2] or on-flow, e.g.  for high-throughput (1H-NMR) monitoring and optimization of reactions.[3] Currently, we are designing NMR-chips for observation also of low-gamma nuclides, for 1D as well as 2D NMR experiments.

[1] R. M. Fratila, A. H. Velders, Annu. Rev. Anal. Chem. 2011, 227-249
[2] M. V. Gomez, D. N. Reinhoudt, A. H. Velders, Small 2008, 4, 1293-1295.
[3] M.V. Gomez, H.J.J. Verputten, A. Díaz-Ortíz, A. Moreno, A. de la Hoz, A.H. Velders, Chem. Commun. 2010, 4514-4516

Dr. Jonathan Williams

Applications of Travelling Wave Ion Mobility Mass Spectrometry in Small Molecule Studies

Jonathan Williams

Waters Corporation

A hybrid quadrupole / ion mobility / oa-ToF mass spectrometer (Synapt G2 HDMS, Waters, UK) has been used in both CID and ETD modes of acquisition. The instrument incorporates three Travelling-Wave SRIG’s prior to the ToF mass analyser. For fragmentation using ETD, a glow discharge source was used to fill the Trap T-Wave cell with quadrupole mass selected ETD reagent anions. During an acquisition, the source polarity and quadrupole set mass are switched to allow multiply charged cations to interact with stored reagent anions in the Trap T-Wave. This interaction allows an ion-ion type reaction resulting in ETD product ions which are separated according to their ion mobilities in the second cell (IMS T-Wave cell). Upon exiting the IMS cell the ions enter the Transfer T-Wave which can be used to transfer ions into the ToF or optionally provide CID prior to the ToF.

A number of diverse application areas will be described. These will include the utility of both CID and ETD combined with ion mobility for some isomeric permethylated oligosaccharides and low molecular weight polymers. Ion mobility has been used to separate two singly charged ion species differing only by the site of protonation of a number of fluoroquinolone antibiotics. The enhanced mobility resolution of the Synapt G2, together with differing drift gas evaluation, has facilitated baseline separation of the ‘protomers’ formed from the fluoroquinolones investigated. A description of how to calculate the CCS values of ions using T-Wave ion mobility will be shown.