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27th Montreux symposium on LC/MS
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27th Montreux symposium on LC/MS

27th Montreux symposium on LC/MS
News

27th Montreux symposium on LC/MS

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The LC/MS technique has become mature as was reflected in the program both by the number of oral (43) and poster presentations (154). A further shift towards applications versus new technological innovation was observed when compared to symposia. However, intriguing new concepts were presented both by university and industrial groups as well as vendors.

The conference was preceded with the traditional two day courses on the basics of “Practical LCMS” and the “Advanced interpretation of CID mass spectra from LC/MS/MS” by Henion and Voyksner and with a half-day course on “LC/MS adequate clean-up of biofluids” by Boos and a full day course on “Metabolomics: state of the art and practical considerations” by Reijmers and Hankemeier. This set of courses provided newcomers the opportunity to build their expertise to a level which enabled them to get the most out of the symposium and the extensive exhibition.

After the opening of the symposium by the chairman Jan van der Greef, the opening “Roland W. Frei” lecture, was given by George Poste. George Poste is the Chief Scientist of the Complex Adaptive Systems Initiative and Professor of Health Innovation at Arizona State University and is recognized as one of the global leaders in Health Care innovation. His lecture addressed the key points that limit current improvements of Health Care systems from a broad perspective as well as from a technical perspective. His excellent presentation is available at www.casi.asu.edu and outlined the key importance of having access to the correct specimen and the standardization of methodologies. In view of the global challenges the aspects of health care costs, demographics, chronic diseases as well as lifestyle diseases were discussed. A shift towards health status monitoring was marked together with new value propositions in health care. Diagnosis is the key focus to observe individual differences and to design new therapeutic options, moving towards personalized health strategies. The importance of biomarkers and biosignatures for these improvements was emphasized, but also the lack of success so far in biomarker research. This is attributed to lack of rigorous validation. Most studies involve reporting of anecdotal studies only. In terms of the analytical validation, the pre-sampling and post-sampling criteria were discussed as well as the performance of current proteomics identifying the current obstacles as observed from lab-to-lab comparison. In addition the opportunities for wireless health status monitoring via in body sensoring based on micro/nano-devices was envisioned. Also the handling of mega-variate data was emphasized as challenge. 

From a nutrition perspective Bruce German in his keynote lecture made clear how prevention will be the focus of the future and he shared the concept of a change of the agriculture/food industry from a cost-driven to a value-driven enterprise as was also proposed for the Health care system by the George Poste. It is clear Pharma and food perspectives are merging in a conceptual sense, it would be ideal to generate a system-wide transformation in our thinking and tackle the health & wellness issues industry-wide. The importance of microbiota in terms of health and the mass spectrometry wish list for the future from the nutrition/agriculture point of view was presented and included quantitation, fast & friendly and annotation of the data at the system level.

Metabolomics as rapid merging field of interest for discovery of biomarkers was evaluated from both a technological as well as a practical, problem solving perspective by Ian Wilson and Thomas Hankemeier. Ian Wilson analyzed the key elements for success; good specimens, standards, study design, technology and validation as introduced by Poste from a pharmaceutical domain. He demonstrated the importance of the microbiome and the importance of understanding the host metabolic phenotype and discussed the possible links with obesity, aging and the option to influence the microbiome via pre-biotic strategies. UHPLC has become a golden standard in metabolomics with HILIC getting more and more attractive for polar metabolites. Thomas Hankemeier elaborated further on the innovative analytical strategies applied in personalized medicine research and in particular how to get the right data spanning the whole metabolomics work flow from the experimental design to the biological interpretation. A calibration and error model per compound in targeted approaches is essential; typically 5-15% reproducibility can be achieved. Identification and understanding biological relevance is mandatory and progress in identification methodology via multi-stage MS by applying fragmentation trees was illustrated. Further refinement and automation of this approach including de-replication via elemental composition has become a first version of this workflow. A round robin test on robustness of MS-tree acquisition demonstrated the protocol to be robust and to be a good basis for structure generation and candidate rejection as well. Miniaturization towards single cell analysis based on micro-and nano-fluidics to create a fully-integrated scale analysis system on a chip appears to be a near-future option. Better coverage, throughput improvements, cost reduction, QC/QA, miniaturization are essential drivers in metabolomics and international collaboration is key to achieve these demanding goals.

Another topic of the conference was surface analysis. Jack Henion overviewed the challenges and illustrated new options via liquid extraction combined with nESI-MS. This method appears to be simple and was shown to effectively profile tissue slices, dried blood spots, TLC spots from stomach contents and wheat. This method is excellent for fast profiling, while MALDI is still the method of choice for high quality imaging as demonstrated for clinical biomarker discovery in breast cancer and immunoimaging in an Alzheimer disease model. Various contributions on this topic including laser-diode thermal desorption/atmospheric pressure chemical ionization tandem mass spectrometry, with no solvent and mobile phase, enabling efficient analysis of phospholipids and dried blood spot with SPE –MS/MS.

Direct analysis in real time (DART) and an Atmospheric Solid Analysis Probe (ASAP) for investigation of surfaces in combination with atmospheric pressure photoionization was reported to have enhanced sensitivity, less fragmentation and potential for extending the range of traceable compounds and reduction of matrix effects. SPME in combination with desorption electrospray ionization (DESI) – MS/MS proved to be a unique tool for rapid analysis of biological samples.

Surface analysis for biological surfaces was the topic of the overview on multi-modal tissue imaging using molecular microscopy given by Ron Heeren. Label free molecular imaging with mass spectrometry provides biomedical tissue imaging of separate compound classes such as metabolites, targeted pharmaceuticals, peptides/proteins or lipids. These images can even be performed at the single cell level. MALDI (photons) enables imaging of larger molecules however with lower space resolution than with SIMS (charged particles) but the latter is limited to lower molecular weight components. Microscope mode imaging (defocusing) and MALDI ion mobility MS imaging are examples of various experimental setups that are currently developed in this field of steep growth in biomedical sciences and in biology as was demonstrated for signaling and development research.

Sampling and preparing whole blood for LC-MS/MS was topic of many presentations and a special expert hearing on this was chaired by Karl-Siegfried Boos. Driven by
ethics 3R (Reduction – Refinement – Replacement of animal models), scientific value, paediatrics (and critically ill patients) and cost savings different approaches were discussed from a pharmaceutical and clinical-chemical perspective both for therapeutic monitoring, non-GLP and GLP based pre-clinical development, clinical studies and diagnostics. Micro-sampling using micropipettes with volumes down to 0.5 µL
was demonstrated to be a powerful approach. Dried blood sampling was shown to become more and more accepted and used in a variety of analytical stages during drug development. Sampling and in-line processing of whole blood for fully automated SPE-LC-MS/MS analysis was explained and conversion of whole Blood (WB) into
Cell-Disintegrated Blood (CDB) via heat-shock treatment or cryogenic treatment was shown to meet the criteria in clinical-chemical analysis, which includes newborn screening, Therapeutic Drug Monitoring (TDM), Point-of-care-testing (POCT), Drug-of-abuse-testing and clinical studies.

On the technology related contributions, besides various new automation procedures including push-button LC-MS/MS approaches, also on the ionization methodology new insights were presented. The main two ionization methods used today in both positive and negative mode are electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI), while for extending the range towards nonpolar analytes atmospheric photoionization (APPI) is expanding. A new flexible ion source with all three options integrated was presented. Furthermore a new chip-based heaterless nano-APCI interface was presented, overcoming several limitations of conventional APCI especially the handling of thermolabile molecules. A field free APCI ion source was presented to be able to operate over a broad flow rate regime from 20 ul – 1.5 L/min.

Developments on the chromatography aspect of LC/MS often related to UPLC but also comprised among others nano ultra performance LC quadrupole TOF-MS, SCX-HILIC-MS for cationic polar metabolites, HPAEC oligosaccharides and ZIC-HILIC enrichment for glycopeptides. On-line electrochemistry MS for exploring phase 1 and II metabolism was demonstrated to be attractive.

Many applications in pharmaceutical, environmental, nutrition biotechnology were demonstrated. Forensic, doping-related and chemical warfare agents detection using GCMS, LCMS and NMR was reported and the use of biomarker detection for alleged use or low-level occupational exposure. For femtomole amount detection of phosphorylated peptides an approach was demonstrated to tackle the problem of incomplete recovery based on ion-mobilizing additives. For biologics/biopharmaceuticals UPLC-MS was also successfully applied for characterization of monoclonal antibodies, recombinant proteins, post-translational modification detection and doping screening. In microbiota related research High Performance Anion Exchange chromatography (HPAEC) in combination with membrane desalting device appeared effective. Furthermore high-resolution lipid profiling for C13 isotope detection in flux studies and porous graphitic carbon (hypercarb) for proteomic studies in oncology demonstrate the broad and in depth applications presented.

The “Roland W. Frei” poster award was on behalf of the poster-award committee presented by Jose Broekaert as chairman of the IAEAC and was granted to Nathanaël Delmotte from the ETH in Zurich for his poster on “Metabolome analysis using nano-scale ion-pair reversed-phase HPLC and high resolution mass spectrometry”.

The seemingly never-ending new inventions and applications in the field of mass spectrometry makes us look forward to the next event in 2014 again in November in Montreux , Switzerland.

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