Day 2 :
Keynote Forum
Elliot R. Bernstein
Colorado State University, USA
Keynote: Euv laser mass spectrometry and photoelectron spectroscopy of mass selected neutral clusters and molecules
Time : 9:00 - 9:30 AM
Biography:
Bernstein received his Ph.D. degree from Caltech and was a post doctoral fellow at the University of Chicago. He has been at CSU since 1975 where he has studied molecular crystal vibrational and electronic excitons and phase transitions, cryogenic liquids, and gas phase clusters with a central focus on intermolecular interactions. Recently his research has focused on chemical reactions of neutral and ionic clusters. This latter research area has involved catalytic and photocatalytic cluster systems, solute/solvent systems, the reactions of ionized molecules and clusters, and initial steps in the release of stored energy molecular species.
Abstract:
A long standing set of goals for studies of systems of inhomogeneous, neutral clusters (e.g., MmXn or (molecule)p) has been to mass sort and select them individually for determination of physical and chemical properties of each neutral cluster by spectroscopic techniques. We have constructed appropriate instrumentation to achieve these important goals employing photoelectron spectroscopy (PES), driven by both visible (for MmXn -) and EUV (for MmXn0) radiation. Our 26.5 eV/photon EUV laser can ionize any neutral cluster or molecule (EUV PES) that can be identified and isolated. The algorithm includes the following steps: 1. generation of cluster negative ions in a laser ablation supersonic source with the addition, as required, of low energy electrons from a Y2O3 disk; 2. separation of these anionic clusters in a reflectron time of flight mass spectrometer (RTOFMS); 3. selection and slowing of specific, chosen clusters in a mass gate/momentum deceleration stage; 4. threshold photo-detachment of the sorted and selected negative ion clusters with a tunable VIS/UV laser to generate neutral, isolated clusters; and 5. EUV PES of these neutral clusters. Such studies generate vibrational and structural information on the ground states of the neutral clusters (through VIS/UV PES), and information on the ion states of the clusters (through EUV PES). The presentation will include PES results on various metal oxides, sulfides, and other cluster systems and molecules.
Keynote Forum
Brigitte Simons
SCIEX, Canada
Keynote: Quantitation of Lipids in High Throughput; the shotgun lipidomics profiling of NAFLD and steatohepatitis by ion mobility-MS data acquisition techniques
Time : 9:30 - 10:00 AM
Biography:
Brigitte Simons is a market development specialist at SCIEX, specializing in metabolomics using accurate mass time-of-flight mass spectrometry solutions. Prior to working at SCIEX, Brigitte received her Ph.D. in Chemical Biology at the University of Ottawa. She then completed two research post-doctoral fellowships at the Centre for Biologics Research at Health Canada and the National Institute of Heart Lung and Blood in Bethesda MD. Brigitte also spearheads an academic partnerships program for our North American business – which listeners can browse sciex.com for more information or contact Brigitte for more information.
Abstract:
This presentation will address technologies that directly address clinical research experimentation of lipids in tissue extracts, keeping sampling and data processing throughput in mind. The human lipidome contains >100,000 different molecular species found within a small mass range; consequently, isobaric overlap makes unambiguous identification and quantitation of lipid species difficult. Herein, methods that utilize high sensitivity MS/MS techniques using a high sensitivity triple quadrupole linked LIT mass spectrometer to isolate lipid classes for identification of molecular composition and quantitation have been investigated. Furthermore, QqQ platforms can now be coupled to differential ion mobility (DMS) devices and have shown to resolve phospholipid sub-classes, triglycerides, and strikingly, the sphingomyelins (SM) were resolved from PC molecular species. The latter observation is significant considering these two classes cannot be resolved using triple quadrupole strategies alone, and their masses overlap significantly. Alterations in hepatic phospholipid composition, especially the amount of phosphatidylcholine (PC) and the ratio of PC to phosphatidylanolamine (PE) might contribute to the development non-alcoholic fatty liver disease (NAFLD). Aim was to compare PC/PE ratio in patients with NAFLD or chronic Hepatitis C (CHC) to healthy controls. This was a cross-sectional study. Liver samples were obtained from healthy controls (HC), patients with simple steatosis (SS), and steatohepatitis (NASH). Lipids were extracted from liver tissue and the total PC and PE lipid detected was quantified by tandem mass spectrometry using multiple precursor ion scanning [normalized by internal standards]. Hepatic PC/PE ratio was lower in all 3 patient groups, mainly due to a reduction in PC, which was significant in SS and NASH compared to controls. This study confirms previous animal data on PC/PE in NAFLD and represents a high throughput lipidomics platform’s utility to clinical outputs.
Keynote Forum
Magnus S. Magnusson
University of Iceland, Iceland
Keynote: Similarity of Hierarchical Structured Clustering in Human and Neuronal interactions and on DNA: Structural and Functional analogies.
Time : 10:00 - 10:30 AM
Biography:
Magnus S Magnusson, Research Professor. PhD in 1983, University of Copenhagen. Author of the T-pattern model and the corresponding detection algorithms in THEME. He has focused on real-time organization of behavior, co-directed DNA analysis, numerous papers and invited talks at numerous conferences and universities in Europe, USA and Japan. Deputy Director 1983-1988, Anthropology Laboratory, Museum of Natural History, Paris. Repeatedly invited temporary Professor at the University of Paris. Since 1991, Founder and Director of the Human Behavior Laboratory, University of Iceland. Since 1995, he is in collaboration between 24 universities based on “Magnusson’s analytical model” initiated at the Sorbonne, Paris
Abstract:
Hierarchical structured clustering (HSC) seems characteristic of the structure of the universe balancing a small number of forces, some pulling others pushing apart, the self-similar fractal distribution of matter in the universe thus reflecting HST rather than just dispersion or clumping. HSC also characterizes a proposed pattern type, called T-pattern, detected in the temporal organization of many kinds of verbal and non-verbal human, animal and neuronal behavior and interactions and is also characteristic of the structure of DNA. Functional analogies seem to exist between the occurrence of T-patterns in “cell city” and in human cities. Structural self-similarity over many levels of biological organization suggests the possibility of a unified (mathematical, bioinformatics and system biological) approach. The T-pattern is described as a repeated hierarchical and self-similar tree structure based on a single non-terminal relation, called a critical interval (CI) relationship. The instances of a T-pattern may be seen as repeated statistical pseudo-fractal objects characterized by statistically significant translation symmetry. THEMEtm (by M.S. Magnusson, ©PatternVision Ltd) is special purpose T-pattern detection and analysis PC software using a special CI detection algorithm combined with an evolution algorithm, presented here together with results from the analysis of behavior and interactions. From the relatively slow time scale of human and animal interactions to the much faster interactions within populations of neurons in living rat brains. Analogies are discussed between T-pattern structure and functions in the „cities of proteins“ (cell city) and human cities, especially regarding specialization and the particular case of religious behavior.
- Track 1: Applications of Mass Spectrometry Track 3: New Approaches in Mass Spectrometry Track 6: Mass spectrometry Imaging Track 8: Ionization Techniques
Chair
Elliot R Bernstein
Colorado State University, USA
Co-Chair
Ioana NUTA
Univ. Grenoble Alpes, SIMAP, France
Session Introduction
Yujun Shi
University of Calgary, Canada
Title: Development of Laser Ionization Methods and Its Application in the Investigation of Chemical Vapor Deposition Chemistry
Biography:
Yujun Shi obtained her PhD in Chemistry from the University of Western Ontraio (now Western University) in Canada. She did postdoctoal work at the National Research Council (NRC) of Canada in Ottawa on an NSERC Visiting Fellowship. She is currently an Associate Professor in the Department of Chemistry at the University of Calgary. She has published more than 50 papers in referred journals and has been serving as an editorial board member for Canadian Journal of Chemistry (Associate Editor) and Frontiers in Physical Chemistry Chemical Physics.
Abstract:
Laser ionization mass spectrometry is a powerful diagnostic tool for a variety of applications including biomedical research, drug delivery, atmospheric chemistry, and chemical vapor deposition chemistry. Our laboratory has developed several complimentary ionization methods, including the non-resonant single photon ionization (SPI) with a vacuum ultraviolet (VUV) laser radiation of 118 nm (10.5 eV) and the laser induced electron ionization (LIEI) for the diagnosis of the decomposition chemistry of precursor molecules in the process of hot wire chemical vapor deposition (HWCVD). In this work, results from our investigation on the decomposition of orgaosilicon molecules on heated tungsten and tantalum wires will be reviewed. A common decomposition pathway of forming methyl radicals has been demonstrated for methy-substituted silane molecules and methyl-substituted four-membered-ring silacyclobutane compounds. The formation mechanism and the activation energy for this pathway will be presented in this work. For the four-membered-ring organosilicon precursors, additional decomposition channels were also found from the ring-opening reactions to form alkene and silenes. The investigation of the decomposition chemistry of organosilicon molecules on heated metal wires provides helpful and important insights in understanding the secondary gas-phase reactions in HWCVD processes and also the surface reactions on the metal catalysts.
Andrew Ewing
University of Gothenburg and Chalmers University of Technology, Sweden
Title: Mass Spectrometry Imaging in Flies, Cells, and Vesicles
Biography:
Andrew Ewing received a PhD from Indiana University. After 25 years at Penn State University, he is now Professor at Chalmers University and University of Gothenburg, Sweden. His 300 publications have been cited over 15000 times with an H-index of 69. He has recently received the Norblad-Ekstrand Medal of the Swedish Chemical Society (2014), and the Pittsburgh Conference Award in Analytical Chemistry (2015). He is an Honorary Professor at Nanjing and Beijing Universities of Science and Technology. He is a member of the Royal Swedish Academy of Sciences (from 2012) and the Gothenburg Academy of Arts and Sciences (2013).
Abstract:
We have been developing mass spectrometry imaging methods to study the process of neurocommunication at the system and cellular level. We focus on PC12 cells as a model of exocytosis and the fly model (Drosophila melanogaster) providing a unique system to examine neurotransmitter release and drug dependence mechanisms in a small, but complete system. Mass spectrometry imaging with ion beams allows spatial resolution of a few micrometers down to 40 nanometers in favorable cases. We have been using secondary ion mass spectrometry (SIMS) with a unique 40-kV argon cluster ion source and the NanoSIMS to measure the lipids across the fly brain and catecholamine in nanometer vesicles, respectively. Here, we have focused on the effect of the drug, methylphenidate, on lipid composition in the brain and find that it varies in a way that might affect learning and memory. We have also used NanoSIMS to measure transmitter in subregions of nanometer vesicles. Combined with other new methods to measure the content of the interior of vesicles, we have begun to investigate the details and implications of open and closed exocytosis on regulation of how the brain works.
Libong Danielle
Paris South University, France
Title: Identification of lipid biomarkers of atopic skin by normal phase chromatography/high resolution mass spectrometry
Biography:
Danielle Libong has completed her PhD at the age of 28 years from Ecole Polytechnique of Palaiseau (France). She is senior lecturer in the Interdisciplinary Academic Unit Lip(Sys)² Lipids (Analytical and Biological Systems) of the School of Pharmacy of Paris-South University. Lip(Sys)² is a research team in analytical chemistry with an important experience in lipid analysis by chromatographic, hyphenated methods and optical spectroscopy. She has published more than 25 papers in reputed journals.
Abstract:
The strategy used to determine the lipid biomarkers involve lipidomics analysis of normal and atopic skin. In the case of skin called atopic, permeability of the epidermis or the stratum corneum is a problem. Stratum Corneum has been described as a type of “brick and mortar”, in which the corneocytes are bricks. Extracellular spaces are filled with lipids that are divided into three main classes: ceramides, fatty acids and cholesterol. Ceramides constitute a major class of lipids and are composed of a sphingoid and a fatty acid moiety. They play a major role in skin permeability. Their organization is essential to maintenance of the barrier function. Lipidomics offers a unique opportunity to analyze the complex role of lipids in cellular processes. The first step focuses on the development of a method for separation and detection of lipid classes molecular species using (Normal Phase) LC/ MS in high resolution. Different ionization modes have been tested between APPI and APCI. The main goal is to collect information from all detected peaks, the profile of the sample provides the relative distribution of species and the molecular mass of each. Statistical comparison of the profiles obtained for several samples of both populations reveal the characteristic signals of over / under expressed signals in each group. In the end we characterize and identify biomarker by MS/MS.
Ioana NUTA
Univ. Grenoble Alpes, SIMAP, France
Title: Mass spectrometry with Knudsen cell applied to the investigation of organometallic precursors vapors
Biography:
Ioana NUTA received her Ph.D from Orleans’s University (France) in 2005 working on fluoride molten salts baths at high temperature by NMR spectroscopy in CNRS-CEMHTI Laboratory. She then completed a post-doctoral fellowship at ENSI Caen (2006) in CNRS-LCS Laboratory and at BRUKER (Germany) in department of NMR probe devellopement for catalytic studies. Since 2007, she joined CNRS- SIMAP Laboratory as reasercher in “Thermodynamic and Process Optimization” team where she has in charge the themodynamic studies on gaseous phase using Knudsen effusion Mass Spectrometery. Current studies focus on thermodynamics of organometallics, molten salts and oxides.
Abstract:
A special Knudsen cell reactor coupled to a mass spectrometer, was specifically designed for the study of organometallic precursors. This reactor is built as tandem cells: an evaporation cell and a cracking cell. This reactor tries to simulate the conditions found in the bubbler of the ALD system by the evaporation cell and in the hot reaction zone of ALD by the cracking cell. The first stage reactor - an evaporation cell - provides an input saturated vapor flow operating from room temperature to 333 K. The second stage cell, named cracking cell, operated from 333 to 723K in the present study. During experiments, the effusion orifice is externally opened for direct mass spectrometric measurements of saturated vapour pressures. The device has been tested using the well-known mercury system. The thermal cracking of the gaseous precursor pentakis dimethylamino tantalum (PDMAT), generally adopted in the ALD/CVD TaN deposition processes, has been studied in the temperature range from 343 to 723K. Experiments showed the apparition of many gaseous species when cracking temperature increased and in particular the dimethylamine, HNC2H6 (g), corresponding to the saturated organic branches of PDMAT. Decomposition products of the HNC2H6 branch were observed at relatively high temperature, namely above 633K. This gas phase study - as for the saturated preceding one- shows the presence of oxygen containing molecules in PDMAT cracked vapor. Thus it allows explaining the systematic presence of oxygen contamination in the deposited TaN films observed in ALD/CVD industrial processes.
Stefanie Maedler
Ontario Ministry of the Environment and Climate Change, Canada
Title: Ultra-trace level speciated isotope dilution measurement of Cr(VI) using ion chromatography tandem mass spectrometry in environmental waters
Biography:
Stefanie Maedler received her Ph.D. in analytical chemistry in 2011 from the Swiss Federal Institute of Technology in Zurich, Switzerland (ETHZ). After 2 postdoctoral fellowships at York University and University of Toronto, Canada, Dr. Maedler joined the Ontario Ministry of Environment and Climate Change as a Development Scientist. She has 10 years of expertise in the use of mass spectrometry (MS) for biological and environmental applications and is currently focusing on the speciation of inorganic analytes and understanding their toxicological effects on fish through MS-based metabolomics approaches.
Abstract:
The reliable analysis of highly toxic hexavalent chromium, Cr(VI), at ultra-trace levels remains challenging, given its easy conversion to non-toxic trivalent chromium. The new approach demonstrates a novel analytical method to quantify Cr(VI) at low ng/L concentration levels in environmental water samples by using speciated isotope dilution (SID) analysis and double-spiking with Cr(III) and Cr(VI) enriched for different isotopes. Ion chromatography tandem mass spectrometry (IC-MS/MS) was used for the analysis of Cr(VI) as HCrO4- → CrO3-. While following a classical linear multipoint calibration curve a method detection limit (MDL) of 7 ng/L Cr(VI) was achieved, the modified SID-MS method adapted from U.S. EPA 6800 allowed for the quantification of Cr(VI) with an MDL of 2 ng/L and provided results corrected for Cr(VI) loss occurred after sample collection. The adapted SID-MS approach proved to yield more accurate and precise results than the multipoint calibration method, allowed for compensation of Cr(VI) reduction during sample transportation and storage while eliminating the need for frequent external calibration. The SID approach permitted continuous sample analysis for several days without the need for recalibration. This new developed IC-MS/MS method represents an alternative to the routinely used inductively-coupled plasma (ICP) instrumentation, IC-ICP-MS, and offers several advantages over detection with ICP-MS for Cr(VI), such as the absence of polyatomic interferences of 52Cr formed in the ICP (36Ar16O, 40Ar12C, 35Cl16OH, and 37Cl14NH) that require the use of dynamic reaction/collision cells or high-resolution double-focusing sector field instruments.
Nhu Phan
University of Gothenburg, Gothenburg, Sweden
Title: Lipid imaging of invertebrate model systems by secondary ion mass spectrometry
Biography:
Nhu Phan has been working on her PhD at the University of Gothenburg, Sweden. Her research is about developing mass spectrometry imaging methods for biological applications on different biological models, from single cell imaging to invertebrate models particularly C. elegans and Drosophila. One of her main researches is studying the effects of administrated stimulant drugs on the neurochemistry of Drosophila brain. The significant finding in this study is that the stimulant drug induces lipid structure of the brain and that the brain lipids could closely relate to learning and memory.
Abstract:
Drosophila melanogaster and C. elegans (flies and worms, respectively) are common biological model systems, which have relatively simple anatomy and behavior but possess highly conserved molecular and cellular processes compared to humans. We have applied time of flight secondary ion mass spectrometry (ToF-SIMS) to study lipid structural effects of stimulant drug methylphenidate on the fly brain, and to investigate the 3D chemical anatomy of C. elegans. Different distributions of various biomolecules, particularly fatty acids, eye pigment, diacylglycerides, phospholipids have been found across the fly brain. Lipid structures, particularly diacylglycerides (DAG), phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositiol (PI), are shown to dramatically alter following the administration of methylphenidate. For C. elegans, the entire worm and worm sections were imaged using 3 dimensional (3D) and 2 dimensional (2D) approaches, respectively. Significant changes in the chemical distribution were observed along the depth of the worm. In addition, correlation between 2D and 3D ion images showed different molecular structures across the worm, possible localization of the nerve ring, the cuticle, and the fluid containing space inside the worm. In addtion, we successfully used tandem MS on ToF-SIMS with a high energy 40 keV Ar4000+ gas cluster primary ion beam (GCIB) to elucidate the structures of molecular lipids in the fly brain and the worm. ToF-SIMS imaging shows great potential to elucidate chemical distributions in small invertebrate systems in relation to endogenous and exogenous effects.
- Track 1: Applications of Mass Spectrometry Track 5: Recent Advances and Development in Mass Spectrometry Track 9: Mass Spectrometry Configurations and Techniques Track 12:NMR Spectroscopy and NMR in biomedicine
Chair
Lianming Wu
GlaxoSmithKline, USA
Co-Chair
Medicharla V. Jagannadham
CSIR-Centre for Cellular and Molecular Biology, India
Session Introduction
Paul Lee
Discovery Technologies, Amgen Inc., USA
Title: High-throughput metabolic stability assays using RapidFire and Accurate-Mass Spectrometry
Biography:
Paul Lee joined Amgen Inc. in 2004. He currently leads a group of scientists and is responsible for developing biochemical and cellular assays, implement high-throughput screens and lead discovery in the early drug discovery process. Prior to joining Amgen, Paul held various positions at Pfizer, Pharmacia, and Glaxo-Wellcome for over 12 years. Paul received his Ph.D. in Pharmacology from University of Hong Kong and completed his postdoctoral training at NIH.
Abstract:
Metabolic stabilities of small molecules, in the presence of liver microsomes and/or hepatocytes, have been frequently assessed during lead optimization in the early drug discovery process. The resultant ranking is a key selection criterion for compound advancement into in vivo studies because a more metabolically stable compound will likely give rise to a longer half-life in animal studies. In vitro metabolic stability studies have been routinely performed using pooled liver microsomes from various species (human, monkey, dog, rat and mouse). The key detection technology of this assay is by monitoring the turnover of the compound in the presence of liver microsomes using liquid chromatography-tandem mass spectrometry (LC-MS). However, the bottleneck of the assay falls on the long separation time in LC. The use of RapidFire, a system consisting of a solid-phase extraction column, automated plate handler, sample injector, and software interface with MS, has significantly improved the throughput. In addition, the use of an Accurate-Mass Quadrupole Time-of-Flight (QTOF) MS system with RapidFire eliminates the need of tuning prior to sample analysis and further streamlines the MS analysis process. In this report, we describe the implement of RapidFire-QTOF system in human and rat metabolic stability assays, resulting in a throughput ten times higher than the traditional LC-MS method without compromising data quality.
Lianming Wu
GlaxoSmithKline, USA
Title: Ambient Mass Spectrometry: New Directions in Pharmaceutical Analysis
Biography:
Lianming Wu obtained a Ph.D. in mass spectrometry (MS) from Purdue University under the guidance of Prof. R. Graham Cooks and followed by Postdoc Research Fellow in MS-based proteomics at Pacific Northwest National Laboratory under the guidance of Dr. Richard D. Smith. Dr. Wu has published forty one peer-reviewed scientific papers and book chapters on the subject of mass spectrometry and gave a number of oral presentations in national/international conferences such as ACS National Meeting and PittCon. He is currently working in Global Spectroscopy, Analytical Sciences & Development, GlaxoSmithKline.
Abstract:
Ambient mass spectrometry (AMS), that allows rapid chemical analysis of untreated samples in the ambient environment, represents novel directions in pharmaceutical analysis. Because AMS is performed by direct sampling/ionization of analytes from native samples, it allows high throughput for the analysis of compounds in matrices using solvent-based desorption methods such as desorption electrospray ionization (DESI) or plasma-based methods such as direct analysis in real time (DART). As one of the most widely studied ambient ionization methods, DESI uses fast-moving solvent droplets to extract analytes from surfaces and propel the resulting secondary microdroplets towards the mass analyzer. The development of AMS takes advantage of the knowledge and experience accumulated in mass spectrometry as well as other areas of chemistry, physics and engineering. Several applications of AMS in supporting pharmaceutical product development are presented: (1) direct analysis of samples in matrix with little to no sample pretreatments; (2) MS-imaging (drug mapping in biological samples and tablet formulation as well as diagnosis of tissues); (3) conformation study of proteins and oligonucleotides; and potentially (4) investigation of salt forms of drugs in tablet or capsule formation. Manipulation of the chemistry accompanying ambient ionization can be used to further optimize chemical analysis. Extensions of the methodology to pharmaceutical analysis (such as direct analysis of biotransformation reaction mixtures in biological matrixes and trace analysis of genotoxic impurities) by reactive DESI via in-situ chemical derivatization (viz. gas-phase ion/molecule reactions) on surface are also covered.
Suneetha Achanti
Department of Pharmaceutical Analysis, Hindu College of Pharmacy, India
Title: PDAS–MSD- LC MS- Prescriptive Drug Analysis Study in Rat Plasma for Multiple Sclerosis Disease by LC-ESI/MS
Biography:
Dr. A. Suneetha, Professor & Head, Department of Pharmaceutical Analysis at Hindu College of Pharmacy, Guntur, A.P since 2001. She has completed her Ph.D at the age of 33 years from Acharya Nagarjuna University, Nagarjuna nagar. She has total 16 years of teaching and research experience. She has published more than 40 research papers in various reputed National and International journals. She served as BOS member of ANU in Pharmacy during 2010 to 2013. She is recognized as Research Supervisor from Acharya Nagarjuna University for guiding PhD scholars and she is a life member of APTI.
Abstract:
Prescribing a single drug and its administration is not sufficient in neuro diseases like multiple sclerosis. Combination therapy is growing enormously to decrease the number of medications for a single disease or their associated diseases. In clinical research estimation of concomitant drugs plays a key role to study the drug-drug interactions. The research in the current article has undertaken to provide an accurate method for evaluate the pharmacokinetic parameters of Carbamazepine (CBZ), Duloxetine (DLX), Tamsulosin (TSL) and Teriflunomide (TFM) using Doxofylline (DXF) and Ibuprofen (IBP) as internal standards (IS) in rat plasma by LC- MS when used as combination therapy. The developed bioanalytical method has been validated according to ICH guidelines. The obtained LODs and LOQs of all the drugs were adequate and may useful to perform the pharmacokinetic study in rat plasma. Based on the results, we can conclude that the present method is suitable for quantification of multiple analytes simultaneously without any interference and matrix effects. The concomitant drug analysis along with the target analyte is more advantageous than single compound analysis and also useful in drug interaction and toxicology studies. This method can also be useful in estimating the plasma samples of patients who administer these drugs.
Sermin Tetik
University of Marmara, Turkey
Title: Mass Spectrometry Technology for Biomarker Discovery
Biography:
Dr. Sermin Tetik is completed her PhD degree at the 2003 years at University of Marmara (Istanbul-Turkey). She have been working an academician at the same University, Faculty of Pharmacy in Istanbul-Turkey as associated professor since five years and project director of her research team focusing on thrombosis -hemostasis area. She has published more than 30 papers and above 50 international abstract in reputed journals and conferences, serving as an editorial board member of repute.
Abstract:
Human tissues and organs have affected depending on pathological condition of diseases. Proteins are basic functioning molecules of cells accordingly to modification intensities. Protein modification can be differences in cellular interaction, localization, activity, protein concentration, and co-/posttranslational. Protein biomarker discovery has covered some subtitles or points such as differantially expressed proteins for disease specific protein isomers and abnormal protein activity. Proteomic analysis and conclusions have allowed the potential protein biomarker discovery and thus diagnosis of disease and prognosis can be possible. MS-based proteomic has correlated between protein modification and a certain disease platform to accurately and confidently measure the proteins in sera or plasma which are low-abundance concentration. MS-based quantitative platform use for tissue samples or several body fluids of protein biomarker discovery. Similar to whole tissue samples human fluids such as gastric fluid, urine, blood have also been evaluated in the last decade studies. Intensity of modifications of proteins as post translational can also be indicative of a disease or its progression. Activity-based protein platform changes have allowed for proteome researchs of enzymatic and protein-drug interaction events.
Matthew B. O’Rourke
University of Technology Sydney, Australia
Title: Improving reproducibility and resolution in Imaging Mass Spectrometry (IMS) through the application of low cost manual sample preparation techniques
Biography:
Matthew O’Rourke currently has his PhD in submission after 2 years of candidature and is a resident at the university of Technology in Sydney Australia. He has published several papers in the field of Imaging mass sectrometry and tissue preservation and is an expert in MALDI applications. He also holds a patent in Australia for the development of proprietary MALDI based technology for the identification of microorganisms.
Abstract:
Matrix assisted laser desorption ionisation imaging mass spectrometry (MALDI IMS) is a technique that is currently seeing widespread use in both research and diagnostics for the spatial analysis of a range of biomolecules. It is currently predominantly used for the diagnostic analysis of cancer biomarkers in tissue microarrays. This requires automated sample preparation methods that need to be highly reproducible. Automated preparation methods rely on the spraying of dissolved matrix that can cause delocalisation of molecules, limiting potential image resolution to that of the smallest possible droplet size (usually >20 µm). The instruments are also expensive to purchase and require regular maintenance and servicing by qualified engineers. We therefore describe an alternate sample preparation method that is both low cost and highly reproducible, utilising sublimation and vapour recrystallisation. This eliminates the issue of delocalisation of molecules and allows for a potential resolution of ~1 µm, thereby enabling highly reproducible, ultra-high resolution images when the mass spectrometry instrumentation that can acquire at sub- µm resolution becomes available.
Medicharla V. Jagannadham
CSIR-Centre for Cellular and Molecular Biology, Hyderabad, India
Title: Chemical modification of peptides followed by mass spectrometry: Implications in proteomics
Biography:
M.V. Jagannadham is working as a senior principal scientist and project leader at the Centre for Cellular and Molecular Biology. He published 45 research papers in internationally reputed scientific journals. He trained several students, conducted meetings and workshops in proteomics. He received “Bharat Jyothi” award from India International Friendship Society, New Delhi in 2014 and “Eminent Mass Spectrometrist” award from the Indian Society for Mass spectrometry (ISMAS) in 2015. His current research Interests are proteomics, particularly in improving the de novo sequencing efficiency of peptides using MS techniques, structural and functional studied of outer membrane vesicles of Gram-negative bacteria.
Abstract:
Determination of the sequence of the peptides, obtained from the proteolysis digestion of a protein, using mass spectrometry, is a crucial requirement in proteomics. Sequence of the tryptic peptides is usually obtained by database search or by de novo sequencing. The poor spectral quality and signal to noise ratio interferes in the analysis. Different types of algorithms (MASCOT, SEQUEST and others) are used for interpretation of the mass spectral data. When the database is not available, de novo sequencing is the only way to determine the sequence. The de novo sequence can be obtained by employing different methods of fragmentation (CID, HCD, ETD and ECD), getting information on their sequence and combining this information. Even with MS instruments with high mass accuracy and speed of analysis, the reproducibility of the mass spectrometry-based proteomics is being questioned from time to time. Acetylation of peptides improved the spectral quality, exhibited by an increase in b ion intensities in the MS/MS spectra, improves the efficiency of de novo sequence and helped in validating the database search results. It is a simple reaction, which can be carried out on complex protein digests as is required in proteomics. The identification of proteins from an Antarctic bacterium Pseudomonas syringae Lz4W and other species using this strategy will be discussed.
Adrijana Leonardi
Jozef Stefan Institute/Department of molecular and biomedical sciences, Slovenia
Title: Snake venomics and antivenomics of the nose-horned viper, Vipera ammodytes ammodytes
Biography:
Adrijana Leonardi is a researcher at the Department of molecular and biomedical sciences at the Jozef Stefan Institute. She is a biochemist with a deep research interest in toxicology, protein structure and structure–function relationships. She has been studying hemostatically active components of snake venoms and other medically interesting compounds from animal venoms. Recently she started collaborations with research groups studying the interactions of nanoparticles with plasma proteins. She is mentoring graduate and undergraduate students. Her work was published in 36 papers in reputed journals.
Abstract:
Snake venoms are a deep well of natural compounds with high specificity for various biological targets, from which we draw new pharmacological leads. The nose-horned viper, Vipera a. ammodytes, is the most venomous European snake inhabiting a large part of the south-eastern Europe and Asia Minor. Hemorrhage and coagulopathies are the most pronounced effects of envenomation in humans. We used snake venomics approach, combining proteomics and transcriptomics, to survey the protein composition of the venom with the aim of discovering new pharmacologically active substances. Proteins were separated by two-dimensional polyacrylamide gel electrophoresis into 208 spots and identified by mass spectrometry using the venom gland cDNA library sequence data. In an activity-guided fractionation, hemostatically active components, effecting blood coagulation and platelet aggregation, were detected and isolated: procoagulants metalloproteases - factor IX, X and prothrombin activators, anticoagulant fibrinogenolytic serine proteinases with kallikrein-like and angiotensin I-degrading activity, C-type lectin-like proteins inhibiting vWF-dependant platelet aggregation, dimeric disintegrins inhibiting vWF-, collagen- and ADP-dependant platelet aggregation and phospholipases A2 with anticoagulant and platelet anti-aggregation activities. The only adequate and efficient treatment of snakebite envenoming is serotherapy. The term “antivenomics” describes proteomic procedure of identification of those polypeptides in the venom which possess epitopes that are weakly or not at all recognized by the standard homologous antivenom. Using antivenomics approach we aimed to deepen and broaden our efforts towards formulation of the optimal antigenic composition of the sample for immunization of animals to prepare highly effective antiserum for immunotherapy after envenomation with the nose-horned viper venom.
Athula B. Attygalle
Center for Mass Spectrometry, Stevens Institute of Technology, USA
Title: Charge-site location and structure elucidation of gaseous ions
Biography:
Attygalle is internationally recognized as a researcher and an educator of mass spectrometry and chemical ecology. He has extensive experience in conducting hands-on laboratory workshops and lectures on mass spectrometry. He has conducted mass spectrometry workshops in Malaysia, Pakistan, Brazil, Costa Rica, the Dominican Republic, Switzerland, Saudi Arabia, Germany, and India. Dr. Attygalle has published nearly 200 peer-reviewed research articles. Research reported in some of his joint publications has been singled out for coverage on numerous occasions by the media including the New York Times. Before joining Stevens, he served as the Director of the mass spectrometry facility of Cornell University from 1988 to 2001.
Abstract:
The location of the charge site is an important prerequisite to interpret fragmentation spectra of ions. Although the challenge appears seemingly simple, in reality it is not at all a trivial problem. One often assumes that the charge location can be easily predicted by knowing the gas-phase acidity or basicity of various groups present in a polyfunctional molecule. However, in reality this a very challenging problem because generalizations valid for solution-based chemistry cannot be extended directly to rationalize gas-phase phenomena. For example, there is sufficient experimental evidence, particularly from ion mobility mass spectrometry, to demonstrate that deprotonated p-hydroxybenzoic acid could exist in gas phase as mixture of carboxylate and phenoxide forms. Analogously, protonated aniline exists as mixture of nitrogen- or ring-protonated forms. Sometimes, the incipient proton that imparts the charge to a molecule is mobile. For example, methyl dihydrocinnamate initially protonates on the carbonyl group. Then the charge ambulates to the phenyl ring before finally migrating to the methoxy oxygen atom to afford a subsequent loss of methanol. Analogoulsy, The charge-imparting proton migrates to the ring before a molecule of water is eliminated from protonated benzoic caid. Using many similar examples, the methods, which include ion-molecule reactions, H/D-exchange studies, ion-mobility spectroscopy, and isotope-labeling experiments, available to elucidate intricate ion-fragmentation pathways will be presented and discussed.
- Track 6: Mass spectrometry Imaging
Session Introduction
Ângelo C. Salvador
University of Aveiro, Portugal
Title: GC×GC-TOFMS-based metabolomics as an approach to study the influence of elderberries (sambucus nigra) ripening and cultivar
Biography:
Ângelo Salvador is a PhD candidate in Chemistry, from University of Aveiro (UA), Portugal. He obtained the Master degree in Biochemistry and Food Chemistry at the same institution. He was co-author of a national patent (PT), 9 international research papers, 1 book chapter. His work was presented in international and national conferences, both oral communications (8) and in poster (11), being one awarded as the best oral communication. He was awarded with the best Master student from Food Chemistry field on UA and the best idea of the 2nd National Prize for SPME. He was part of a finalist team (8 out of 500) National Entrepreneurship Prize.
Abstract:
Great challenges on analytical chemistry and mass spectrometry were faced with the expansion of plant metabolomics due to the metabolites complexity. Volatile terpenic (C10 and C15) and norisoprenoid (mainly C13) metabolites, in particular, have similar structural backbones being the MS analysis often difficult. GC-MS is commonly used to analyse these compounds but the demands of plant complexity frequently overwhelm the capacity of 1D separation processes, leading to chromatographic co-elutions and limiting reliable MS identifications. Thus, this work presents a GC×GC-ToFMS-based plant metabolomics approach that explores the effects of ripening and cultivar on elderberries volatile terpenic and norisoprenoid metabolites. GC×GC-ToFMS was selected due to its high resolution and sensitivity being attained by employing an orthogonal separation mechanism (volatility and polarity). Further, ToFMS analyzer granted a high spectral resolution with narrow peaks (milliseconds) and spectral acquisition rates (100Hz). The structured chromatogram (compounds structurally related were positioned on similar chromatographic spaces) combined with the ion extraction chromatography increased the method specificity and sensitivity, helped to eliminate hundreds of non-target metabolites and classify unknown compounds. Sixty-six metabolites were identified being 48 reported for the first time as elderberries components. By employing chemometric tools it was observed that ripening was the variability main factor (39.8%), followed by cultivar (10.3%).
Meryem SeferinoÄŸlu
Sarayköy Nuclear Research and Training Centre, Turkey
Title: Evaluation of uncertainty components associated with alpha-particle spectrometric measurements of plutonium isotopes in bilberry
Biography:
Meryem Seferinoğlu has completed her PhD at the age of 33 years from Hacettepe University, Institute of Pure and Applied Sciences, Department of Chemistry (Turkey). She worked as a postdoc at Luleå University of Technology, Division of Physics & Division of Process Metallurgy (Sweden) and worked as a researcher at European Commission-Joint Research Center-Institute fort reference Material and Measurements. She has been working Turkish Atomic Energy Authority, Sarayköy Nuclear Research and Training center, Radionuclide Metrology Department since 2007.
Abstract:
The measurement uncertainty is an essential part of the assessment of the measurement result. Previously the uncertainty in the radioactivity measurements was defined by the standard deviation of repeated measurements or was not reported. The standard deviation in a radioactive measurement given by the square root of the number of counts reflects only the statistical uncertainty of counting. Recently, more attention is given to the evaluation of the measurement uncertainty and to the preparation of uncertainty budget. There is, however, no common procedure for the estimation of a realistic uncertainty. The ISO Guide “Guide to the Expression of Uncertainty in Measurement†sets up general rules for evaluating and expressing uncertainty in a physical measurement. Some recent European documents give further details about calculating uncertainty in calibration and in quantitative chemical analysis. Nonetheless, the application of these concepts to environmental radioactivity measurements involving radiochemistry is not as straightforward as it should be, because the evaluation of uncertainty requires the analyst to take into account all stage of the method and all possible sources of uncertainty. The main purpose of this study is to demonstrate a procedure for evaluation of the measurement uncertainty in the analysis of plutonium isotopes in bilberry using a radiochemical separation method and alpha-particle spectrometry. The steps in the analytical procedure that contribute considerably to the combined measurement uncertainty are also identified. Identification of these steps might help lower the overall uncertainty of the measurements through decreasing the uncertainties in these steps.
RuAngelie Edrada-Ebel
University of Strathclyde, United Kingdom
Title: A metabolomics approach to monitor the production of secondary metabolites in microbial symbionts and endophytes through automated dereplication of HRFTMS data
Biography:
RuAnglie Edrada-Ebel is a senior lecturer in Pharmaceutical Analysis at the Strathclyde Institute of Pharmacy and Biomedical Sciences in Glasgow since 2008. She was educated a pharmacist and received her PhD in Pharmaceutical Biology from the University of Würzburg in Germany in 1998, followed by a postdoctoral fellowship at the University of California at Santa Cruz. She heads the Natural Products Metabolomics Group at the Strathclyde Institute of Pharmacy and Biomedical Sciences and leads a series of on-going projects on the application of metabolomics to identify and biotechnologically optimize the production of bioactive secondary metabolites in marine-derived microorganisms.
Abstract:
High resolution Fourier transform mass spectrometry (HRFTMS) was employed as complimentary metabolomic tool to dereplicate chemical profiles of sponge-associated microbes. Sponges act as hosts to a diverse population of symbiotic organisms including bacteria and fungi. These symbionts offer benefits to the host including protection, nutritional benefit and support to the sponge skeleton. They have also been recognised as an important source of secondary metabolites which may have bioactivity and therefore medicinal potential. The innovative strategy involved targeted cultivation and isolation of biologically active compounds. Principal Component (PCA), Hierarchical Clustering (HCA), and Orthogonal Partial Least Square-Discriminant (OPLS-DA) analyses were used to evaluate HRFT mass spectral data of culture extracts. The results of the statistical analysis identified and validated the best culture conditions and extraction procedure which optimized the isolation of novel bioactive metabolites. Production of secondary metabolites were investigated in several of the bacterial symbionts that were isolated from marine sponges. Novel secondary metabolites were screened using high resolution mass spectrometry-based metabolomics approaches. Metabolomic profiling using HR-ESIFTMS-MS fragmentation were done at different stages of the growth phase for both solid and liquid culture media and subjected to molecular networking. Molecular networking the mass fragmentation data pinpoints functionalities that can be correlated to bioactivity and fermentation parameters as well as to predict and enhance the biosynthetic pathway involved in the production of the target secondary metabolite. Dereplication studies were accomplished by utilizing the Mzmine software with Antibase and DNP databases. The optimised method in terms of media, incubation time, and maximum production bioactive compounds are taken into account for the scale-up. Metabolomic- and bioassay-guided isolation approaches were carried out to target the compound(s) of interest. We apply a similar approach on terrestrial endophytic fungi. Endophytic fungi also represent a potential source of novel chemistry and biology.
R.K. Vatsa
Bhabha Atomic Research Centre, India
Title: Photoionization of xenon clusters: Does the increasing laser intensity help in multiple ionization ?
Biography:
Dr. R.K. Vatsadid his M.Sc. (Physical Chemistry) from Delhi University and obtained his Ph.D. under supervision of Dr. J.P. Mittal from University of Mumbai, India followed by postdoctoralstudies (Prof. J. Wolfrum) at Heidelberg University, Germany. He is currently Heading Structural ChemistrySection in Chemistry Division of BARC. He has published 140 research papers in internationally reputed journals. He is vice-president of Indian Society for Mass Spectrometry (ISMAS) and an elected fellow of National Academy of Sciences (NASI), India
Abstract:
In the photoionization of CH3I clustersby laser pulses with an intensity of 109 W/cm2 (ï¬= 532 nm), formation of energetic, multiply charged atomic ions was reported by us[1].The C+3 ions,detected in TOF-MS,require a total energy of about 100 eV for formation (starting from CH3I) and in addition they had kinetic energy of ~ 300 eV. Increasing the size of the cluster resulted in enhancement of charge state as well as kinetic energy of the ions. Similar observations were made for other molecular as well as atomic cluster systems. For a given average cluster size, a further enhancement in charge state can be achieved by increasing the intensity of ionising laser. In the case of atomic clusters, we have previously reportedformation of multiply charged xenon ions up to Xe+10 state using 8 ns, 532 nm laser pulses with anintensity of~ 5x 109 W/cm2 [2]. We now report our results on ionization of xenon clusters (of similar average size) using 30 ps pulses with an intensity of ~1012 W/cm2. The ionization wavelengths(266 and 532 nm) were selected to probe if wavelength plays any role in multiple ionization. For the case of high intensity ionization at 266 nm, a strong mass peak for Xe+, followed by a mediumintensity peak for Xe+2 and a small peak for Xe+3 were observed. For these three charged states, theabundance of given state follows Boltzmann distribution. For photoionization at 532 nm, charge state up to Xe+10 could be observed with Xe+9 being the most abundant ionic species.For the lower charge states (+2 to +7), a non- Boltzmann distribution of ionic states was observed. The main cause of such a behavior is absorption of high amount of energy from the laser pulse leading to extended depletion of lower charge state which act as feedfor higher charged states.Theseresults at 1012 W/cm2 intensity once again confirm previous observations that the total energy absorbed by the cluster is much higher at longer wavelength. During the presentation, results obtained on photoionization of Xenon clusters at 1012 W/cm2 will be compared with earlier results obtained using 109 W/cm2. The apparent absence of intensity effects on overall ionization state of xenon will be discussed taking into considerationinitial multiphoton ionization by the laser pulse and subsequent electron induced ionization.
- Track 11: Advances in Chromatography
Session Introduction
Franco Biasioli
Fondazione Edmund Mach, Italy
Title: Endogenous and exogenous volatile compounds in breath as detected by PTR-MS
Biography:
Franco Biasioli holds a degree in Physics (University of Trento, Italy) and a PhD in Physics (University of Innsbruck, Austria). He is a senior researcher at Fondazione Edmund Mach (Italy) where he set up and leads a facility for the high-sensitivity monitoring of volatile organic compound based on direct injection mass spectrometry and spectroscopic methods. His research is focused on food applications but extends also to health and environment. He has published more than 140 papers in ISI journals (h-index=26, WoS) and is serving as a reviewer both for reputed journals and agencies.
Abstract:
Among the different approaches to volatile organic compound (VOC) profiling, Proton Transfer Reaction Mass Spectrometry (PTR-MS), shows intriguing peculiarities: it is fast, non-invasive and can provide very high sensitivity. Moreover, recent technological advancements partly overcome its limitations in compound identification and quantification. Here, the use of PTR-MS for the real time measurement of the concentration of VOCs in breath is discussed with reference to two studies: i) the monitoring of exogenous flavor compounds during food consumption and ii) the monitoring of endogenous compounds as possible markers of diseases or diet. In the first study the nose-space concentration during coffee drinking has been compared with dynamic sensory methods to investigate the perceived effect of different roasting. Data indicates that nose-space concentration can be used to distinguish different coffee samples and different assessors and to identify markers of sensory quality which clearly show multimodal characteristics. In the second series of studies the breath of human and rats has been measured to assess whether VOC concentration can be related to diseases (cirrhosis and coeliac disease) or diet. Data indicates that markers of cirrhosis can be identified by breath analysis and that they are correlated with serum bilirubin while no evidence of breath markers can be found for coeliac disease if the patience follow a gluten free diet. In conclusion: PTR-MS allows the real time analysis of breath VOCs and can be used both for exogenous or endogenous compounds analysis to investigate flavor perception or markers of disease or diet.
Biography:
Dr. Nita-Lazar received her Ph.D. in biochemistry in 2003 from the University of Basel for studies performed at the Friedrich Miescher Institute for Biomedical Research, where she analyzed protein glycosylation using mass spectrometry methods. After postdoctoral training at Stony Brook University and Massachusetts Institute of Technology, where she continued to investigate post-translational protein modifications and their influence on cell signaling, she joined the Program in Systems Immunology and Infectious Disease Research, now the Laboratory of Systems Biology, in April 2009.
Abstract:
Chemotaxis and toll-like receptor (TLR) signaling in macrophages are critical to the immune response. To model these signaling networks, we are using selected reaction monitoring (SRM) to measure the absolute abundance of pathway proteins, and the resulting values are being used as pathway model parameters. RNA-seq was performed to identify expressed transcripts, and shotgun mass spectrometry was used to identify proteotypic peptides. SRM using heavy-labeled internal peptide standards was used to quantify the chemotaxis pathway proteins (1). The transcript and protein abundance values correlated strongly, and estimated protein abundance values for the entire macrophage proteome were produced. Computational modeling of the chemotaxis pathway was performed using Simmune toolset. Molecular reaction rates were constrained using a model training dataset consisting of multiple types of in vitro microscopy data. Subsequently, model testing used an orthogonal dataset from ELISA assays of RAC1-GTP and RHOA-GTP. The model produced in silico results consistent with both the training and testing datasets, and it was determined to be robust by assessing the accuracy of thousands of perturbed models (2). These findings demonstrate the feasibility and value of combining mass spectrometry-based measurements with pathway modeling for advancing biological insight. SRM assays for the canonical TLR signaling pathway and related proteins have been also been successfully developed. A preliminary model of the TLR pathway has been developed using the resulting estimated protein abundance values. The relative proteome and secretome measurements for cells stimulated with TLR ligands and the cytokine secretion values measured by flow cytometry and ELISA are being used as additional constraints for this complex model. This research was supported by the Intramural Research Program of the National Institute of Allergy and Infectious Diseases, NIH.
Namrata Singh
Indian Institute of Chemical Biology, India
Title: Fibrinolytic activity in bovine bile lipid: Application of chromatography, mass spectrometry and In-vivo wound healing assay.
Biography:
An ether extract of nine different bacterial metabolites combined with two step (ether followed by ethanol) extract of bovine bile lipid is used as an immune stimulatory drug. While characterizing the drug, we observed fibrinolytic activity in the extract through fibrinogen plate assay and fibrin zymography. Background literature emphasized major role of fibrinolytic enzymes in activating immune systems. This increased our curiosity to understand the role of these enzymes in this drug in human physiology. This fibrinolytic enzyme/s has no similarity with plasmin in terms of cross reactivity in immunoblot assay and hydrolysis of the specific substrate S-2251. In RP-HPLC analysis, the lipid extract was fractionated into several components. Interestingly, fibrinolytic activity was confined to all the fractions. To purify the enzyme, it was extracted from the lipid by aqueous buffer extraction and applied to CNBr activated fibrinogen substrate affinity column. Purified enzyme was tested for activation of complement system and wound healing through C3 binding and in-vivo wound healing assay respectively. The enzyme will be identified by mass-spectrometric analysis. Also, we propose to finger-print protein components present in bile lipid by MS analysis to have a better insight of the functionality of the lipid component of the drug.
Abstract:
Namrata Singh is an INSPIRE fellow of the DST (India). She is currently associated with CSIR-Indian Institute of Chemical Biology, Kolkata aiming for Ph.D. from the University of Calcutta and will submit her thesis soon. She is involved in characterizing an immune-stimulatory drug. She participated in the 7th HOPE meeting with Nobel Laureates as JSPS Fellow in Japan, 2015. She is a recipient of several national awards and also been felicitated for excellence in academic background. Publication of 4 papers with good citations has initiated her research career and a reviewer of applied medical research.
Somenath Ganguly
Sun Pharmaceutical Industries Ltd, India
Title: Hyphenated Mass Spectrometric techniques: Revolution in Pharmaceutical Analysis
Biography:
Somenath Ganguly has completed his PhD at the age of 28 years from Indian Institute of Technology, Kharagpur. He is the Associate director in the Analytical Research Department of Sun Pharmaceutical Industries Limited, Gurgaon. Sun Pharma is a premier generic pharmaceutical organization. He has published more than 10 papers and patents. He is having about twenty years of experiences in the pharmaceutical analytical research and responsible for solid state characterization, impurity profiling and chromatographic method development activities. He has vast experiences in regulatory and quality requirements in the pharmaceutical industries
Abstract:
In recent days hyphenated mass spectrometric techniques are becoming an essential tool in generic pharmaceutical analysis. United States and European pharmacopeia recommends methods in the general chapters and monographs. Techniques involve are LCMS/MS; GCMS; ICP MS; TGA MS etc. ICH; OSHA, EMEA , US FDA published guidelines for assessing genotoxic and elemental impurities in the active pharmaceutical ingredient( API) and finished drug products. Genotoxic impurities involves structures like Epoxides; Alkylating agents; Aldehydes; Nitro compounds; Hydrazines; Azo compounds etc. Elemental impurities arising out of the catalysts used in the API synthesis like palladium charcoal; nickel acetate, borohydrides; metal residues from reactors line like lead, arsenic, mercury and cadmium. The quantitation levels of these impurities are determined based on maximum daily intake of the drugs and in general limits are in the range of less than 50 ppm. Analytical methods need to be developed, adequately validated and transferred to QC locations as per ICH and regulatory requirements. Challenges and procedures in the analytical method development and validations process involving hyphenated mass spectrometric techniques will be discussed.
A. C. Sahayam
Bhabha Atomic Research Centre, Hyderabad
Title: Determination of cobalt compounds in functional drinks and nutritive supplements using HPLC-ICP-MS
Biography:
Dr. A. C. Sahayam completed his PhD at the age of 37 years from Osmania University, Hyderabad (India) and Postdoctoral research work at National Sun-Yat Sen University, Kaoshiung (Taiwan). He is Head, Bulk Analysis Section at National Centre for Compositional Characterisation of Materials, Bhabha Atomic Research Centre, Hyderabad. His field of expertise is analytical atomic spectrometry using ICP-OES and ICP-MS coupled with different sample introduction techniques. He has published more than 50 papers in reputed journals and serving as a reviewer for reputed journals.
Abstract:
Speciation analysis of cobalt in nutritive supplements has been carried out using HPLC and ICP-MS equipped with a membrane desolvation sample introduction system as detector. In this study, cobalt containing compounds, namely Co(II), cyanocobalamin (CN-Cbl), hydroxylcobalamin (OH-Cb1), 5’-deoxyladenosylcobalamin (Ado-Cbl) and methylcobalamin (Me-Cbl), were well separated by reversed phase HPLC with a microbore C8-HPLC column as the stationary phase. Gradient elution using ammonium acetate and acetonitrile at pH 4 allowed the chromatographic separation of all species, including strongly adsorbed Ado-Cbl and Me-Cbl, in less than 14 minutes. Detection limit was in the range of 0.007-0.031 ng Co mL-1 for various Co species. We determined the concentrations of cobalt containing compounds in selected nutritive supplements and two functional drinks. Over 91% of the total cobalt species was extracted using a 0.5% v/v HNO3 solution in a microwave field at 70oC within a period of 10 min; and the spike recovery was in the range of 94-101% for various species. The HPLC-ICP-MS results showed a satisfactory agreement with the total cobalt concentrations obtained by ICP-MS analysis of functional drinks and dissolved nutritive supplements samples.
Mohammad Azharuddin
University of Calcutta, India
Title: Functional Proteomic Approach in Dry Eye Disease
Biography:
Mohammad Azharuddin has submitted his PhD at the age of 28 years from Department of Biochemistry, University of Calcutta. He has published 6 papers in reputed journals and 3 patents filed under his name.
Abstract:
Identifying important biomarker(s) for a disease is a major challenge in clinical proteomics. In this paper we have introduced a metadata approach to classify proteomes from aqueous deficient dry eye (ADDE) disease. After initial selection of tear samples from control and ADDE subjects a disease specific proteome set was constructed using a simple set theory based reduction method. The properties for each of such protein members were then evaluated, and the properties having highest discriminatory power (between control and ADDE) were chosen. The electrostatic charge distribution and amino acid composition and to some extent overall hydrophobicity seemed to be the important discriminatory features. The ADDE patients thus showed a positive bias in their proteome as compared to control subjects. Similarly, the disease specific proteome contained lesser percentage of glycine and higher percentage of threonine. The meta-proteomic analysis thus implied that ADDE specific proteins have lesser flexibility in their conformation. The results seemed to have some correspondence with our recent finding that ADDE is a protein conformational disease having higher propensity for occurrence of aggregated proteins.
Bernhard Welz
Federal University of Santa Catarina, Brazil
Title: High-temperature high-resolution molecular absorption spectrometry – a supplementary technique to ICP-MS for the determination of halogens.
Biography:
Bernhard Welz has completed his PhD at the age of 29 years from University of Stuttgart (Germany). He worked for Perkin-Elmer for 32 years, the last 16 as Director of Applied Research. After his retirement, he went to Brazil and worked as visiting professor at several Universities. Now he is voluntary professor at the Federal University of Santa Catarina. He has published more than 280 papers in reputed journals with more than 9600 citations; he has an H-index of 54.
Abstract:
Inductively coupled plasma mass spectrometry (ICP-MS) is without doubt one of the most sensitive techniques for the determination of trace elements in a variety of matrices. However, there are also some limitations, and the most serious ones are with the determination of the halogens. Fluorine, which has an ionization energy of 17.42 eV, can essentially not be determined by ICP-MS, as the ionization energy of the plasma gas argon is only 15.76 eV. Chlorine and bromine have lower ionization energies of 12.97 eV and 11.81 eV, respectively, and can be determined by ICP-MS; however, severe matrix and memory effects can often be observed when conventional sample introduction with a nebulizer/spray chamber is used. One way out of this problem is using electrothermal vaporization (ETV) for sample introduction. High-temperature high-resolution molecular absorption spectrometry (HR-CS MAS) is ideally suited for the determination of non-metals, and can therefore be used as a supplementary technique to ICP-MS. The sample – in many cases a solid sample – is directly introduced into a small graphite tube furnace, a ‘molecule-forming reagent’ is added, and the furnace is heated to a previously optimized temperature – typically between 1500 °C and 2500 °C. At this temperature, diatomic molecules are formed in the gas phase, such as CaF, SrCl or CaBr, which exhibit a characteristic absorption spectrum, which can be used for the sensitive determination of these halogens. Plant materials, coal, copper concentrate, cosmetics and fish oil were investigated with this technique and the results compared with those of ICP-MS where possible.
Marek M Kowalczuk
University of Wolverhampton, UK
Title: New vistas in multisatge mass spectrometry for analysis of biodegradable polymers
Biography:
Marek M. Kowalczuk received his Ph.D. degree in 1984 from the Faculty of Chemistry, Silesian University of Technology, and D.Sc. degree in 1994 at the same University. He was a visiting lecturer at the University of Massachusetts in Amherst, MA, U.S.A. in 1990 and Marie Curie EU fellow at the University of Bologna, Italy. Currently, he is professor at the University of Wolverhampton, UK and at the Centre of Polymer and Carbon Materials, Polish Academy of Sciences, Zabrze, Poland. He is the author and co-author of over 130 scientific papers and a score of patents.
Abstract:
Biodegradable polymers play an important role in human life, increasingly irreplaceable one. Knowledge on the relationships between their structure, properties and function is essential for prospective applications of such materials in the areas safe for human health and environment. When the development of biodegradable polymers was in its infancy the most crucial features were concentrated on the effect of macromolecular architecture, new monomer systems, polymerization mechanisms and different polymerization techniques on final biodegradable properties. Significant efforts have been directed towards specific areas, such as mechanisms of biodegradation, biocompatibility, processing conditions and potential applications in medicine, protection of environment and agro chemistry. However, such aspects like bio-safety of biodegradable polymers or nano-safety of their composites were and still are frequently neglected. In the endeavor to safe biodegradable polymers mass spectrometry methods are of particular importance in (co)polymers analyses due to their high sensitivity, selectivity, specificity and speed. Examples of the mass spectrometry studies for sequencing of biodegradable (co)polymers with the use of multi-stage electrospray mass spectrometry (ESI-MSn) will be presented. The special emphasis will be given to the ESI-MSn applications in the synthesis of biodegradable copolyesters as well as ESI-MSn for identification of selected biodegradable polymers on the way of molecular labeling. The attempts to solve the difficult question regarding the molecular structure of biodegradable copolymers with relation to the specific area of applied research will be also discussed.
SÃlvia M. Rocha
Universidade de Aveiro, Portugal
Title: Metabolomics workflow construction for Aspergillus niger detection using advanced gas chromatography
Biography:
SÃlvia M. Rocha is assistant professor in the Chemistry Department at the Aveiro University. She is BA in Pharmaceutical Sciences (Coimbra University), and PhD in Chemistry (Aveiro University). For the last 24 years, her research was focused on the characterization of plant-derived products, prospection of bioactive compounds, and metabolomics applied to bodyfluids, plants and microbial systems. Her main skills are oriented to sample preparation and high through-put analysis based on comprehensive two-dimensional gas chromatography and mass spectrometry. She is leader of the x-Chromatography Lab (goo.gl/gHKsAi) and published over 95 SCI papers, 2 books, 7 book chapters, and 3 patent applications
Abstract:
Opportunistic infections by Aspergillus niger have increased in the last years, either in paediatric patients as adults, presenting a high mortality rate, therefore strongly suggesting the need for prevention or earlier diagnosis and treatment. Microbial metabolomics has been breaking new ground as very useful tool in several areas, including those related to microbial detection, since microorganisms produce several volatile metabolites that can be used as unique chemical fingerprints of each species, and possibly of strains. This richness of information holds the promise for diagnosing infections in situ (e.g. from body fluids, food products, environmental samples, among others), circumventing the laborious recovering of microbes or their genetic material. Microbial metabolomics studies have been mainly focused on the study of the volatile fraction by using 1D-GC. Nevertheless, the use of comprehensive two-dimensional gas chromatography (GC×GC) has revealed that sensitivity and limits of detection are improved compared to 1D-GC. Several challenges should be overcome, since microbial culturing in representative conditions, alongside the technical difficulties to identify and/or quantify trace metabolites within complex matrixes, as well as the inherent problems related to data processing are partially responsible for the paucity of information on the full volatile metabolome of common microbial pathogens. Thus, this talk aims to discuss new developments towards the establishment of a comprehensive platform for A. niger detection management, contributing to in-depth explore its exometabolome, which was studied upon different growth conditions, using a methodology based on headspace-solid phase microextraction combined with GC×GC-ToFMS, an advanced gas chromatographic based methodology with high resolution and high throughput potentialities. Partial Least Squares-Discriminant Analysis (PLS-DA) and cross validation were performed to assess both the predictive power and classification models robustness. In addition, PLS-DA-Variable Importance in Projection was applied to highlight the metabolites playing major roles in species distinction; decreasing the initial dataset to only 16 metabolites (A. niger Biomarker pattern). The data pre-processing time was substantially reduced, and an improvement of quality-of-fit value was achieved. This study goes a step further on exploring the potentialities of metabolomics for constructing A. niger omics pipeline that can be proposed as a high throughput tool towards its future detection based on a molecular biomarkers pattern.
Lee Suan Chua
Universiti Teknologi Malaysia, Malaysia
Title: Protein from edible bird’s nest samples by mass spectrometry
Biography:
Chua Lee Suan has completed her PhD in 2005 from Universiti Teknologi Malaysia. She is a registered chemist, university lecturer and team leader of research group in the analytics and validation study on natural products, particularly for metabolite profiling of herbs and honey. Till to date, she has published more than 50 international research articles and serving as editor for repute journals such as Journal of Ginseng Research and European Journal of Medicinal Plants.
Abstract:
Edible bird’s nest (EBN) is a delicacy and nutritious salivary secretion of swiftlets. It contains mostly protein and carbohydrate, and fat as a minor component. The proteome of EBN is still intensively investigated, mainly due to the recent scientific findings on its medical benefits and rejuvenation of skin complexion. In the present study, proteins from EBN samples were extracted by ultrasonic and detergent assisted techniques, as well as buffer solublization. Aqueous extraction assisted by ultrasonic energy produced the highest protein content, followed by detergent assisted method and buffer solubilization. A wide range of protein sizes ranging from 17-150 kDa, mostly water soluble proteins were detected from EBN samples based on gel electrophoresis. EBN from different locations were found to have slight variance in their electrophoretic protein profile. This could be due to the difference in food sources for swiftlets. The use of detergents such as non-ionic Triton X-100 and ionic SDS produced additional protein bands which could be membrane or transmembrane proteins. The trypsinized proteins were subjected to LC-MS/MS analysis and the mass spectra were matched to the protein database. Acidic mammalian chitinase precursor is the most abundant protein in EBN. The protein was reported to have anti-inflammatory activity against allergy and airway diseases. There are also collagen, lysyl oxidase homolog 3 and co-enzyme Qbinding proteins which are important for skin complexion. The results could explain the remarkable findings of previous investigators on the benefits of EBN consumption. These proteins are first time reported to be presence in EBN.
Marek M. Kowalczuk
University of Wolverhampton, UK
Title: New vistas in multisatge mass spectrometry for analysis of biodegradable polymers
Biography:
Marek M. Kowalczuk received his Ph.D. degree in 1984 from the Faculty of Chemistry, Silesian University of Technology, and D.Sc. degree in 1994 at the same University. He was a visiting lecturer at the University of Massachusetts in Amherst, MA, U.S.A. in 1990 and Marie Curie EU fellow at the University of Bologna, Italy. Currently, he is professor at the University of Wolverhampton, UK and at the Centre of Polymer and Carbon Materials, Polish Academy of Sciences, Zabrze, Poland. He is the author and co-author of over 130 scientific papers and a score of patents.
Abstract:
Biodegradable polymers play an important role in human life, increasingly irreplaceable one. Knowledge on the relationships between their structure, properties and function is essential for prospective applications of such materials in the areas safe for human health and environment. When the development of biodegradable polymers was in its infancy the most crucial features were concentrated on the effect of macromolecular architecture, new monomer systems, polymerization mechanisms and different polymerization techniques on final biodegradable properties. Significant efforts have been directed towards specific areas, such as mechanisms of biodegradation, biocompatibility, processing conditions and potential applications in medicine, protection of environment and agro chemistry. However, such aspects like bio-safety of biodegradable polymers or nano-safety of their composites were and still are frequently neglected. In the endeavor to safe biodegradable polymers mass spectrometry methods are of particular importance in (co)polymers analyses due to their high sensitivity, selectivity, specificity and speed. Examples of the mass spectrometry studies for sequencing of biodegradable (co)polymers with the use of multi-stage electrospray mass spectrometry (ESI-MSn) will be presented. The special emphasis will be given to the ESI-MSn applications in the synthesis of biodegradable copolyesters as well as ESI-MSn for identification of selected biodegradable polymers on the way of molecular labeling. The attempts to solve the difficult question regarding the molecular structure of biodegradable copolymers with relation to the specific area of applied research will be also discussed.
Nuran Elmacı
İzmir Institute of Technology, Gülbahçe-Urla, Turkey
Title: The Structural Analysis of b5 Type Mass Fragment Ions
Biography:
Nuran Elmacı is professor in İzmir Institute of Technology, Gülbahçe-Urla, Turkey
Abstract:
Tandem mass spectrometry (MS/MS) coupled with collision-induced dissociation (CID) is commonly used method for proteomic studies. Upon low-energy CID, protonated or multiply-protonated peptides undergo fragmentation via cleavage of amide bonds to generate N-terminal b and a and/or C-terminal y ions. Understanding the gas-phase structures of these ions and their fragmentation reactions has played a vital role for correct and reliable peptide/protein identification. It has been shown that, bn (n=2-4) ions commonly have oxazolone structure. However, for some cases b2 ions also adopt a six-membered ring; diketopiperazine structure. The formation of these two different structures entirely depends on the amino acid composition of the peptide sequence as well as peptide chain length. The macrocyclic structure of b5+ and larger b ions has been predicted for aliphatic peptides in experimental and theoretical studies. The macrocyclic structure is a problem for determining the amino acid sequence of a peptide. The memory of original primary structure is lost if reopening of the macrocycle takes place at different sites on the cycle. In such cases, the databases of computer programs which are used with the MS/MS results will be insufficient to identify peptide/protein. These databases must be improved by the detailed and comprehensive studies of the gas phase fragmentations of protonated peptides. This will lead to get reliable results for the proteomic researches. In this work, the structures of b5 ions; XA4 composed of alanine and X= X; Asn, Asp, Leu, Phe, Tyr, Cys, amino acid residues have been investigated by using molecular dynamics simulation method and quantum chemical methods. The X-residues have different type of substituent groups; aliphatic (-CH2-), aromatic (C rings), alcohol (-OH), thiol (-SH), amides (-CONH2) and carboxyl (-COOH). To examine the position influence of X residue, three types of linear peptide sequences have been used in the computational calculations for b5+ fragment; XAAAA, AAXAA, AAAAX. It has been observed that the b5 ions prefer to have a macrocyclic structure rather than linear oxazolone isomer. The macrocycle has some common features (backbone structure, proton location, H-bond etc.) for all b-ions studied in this research. The results of this work will be useful to understand the gas phase dissociation chemistry of peptides and will support and insight to the experimental studies.
Yuri E. Moskalenko
Russian Academy of Sciences. Russia
Title: Spectrum analysis applied for the study of intracranial slow volume fluctuations, recorded by coupling of noninvasive methods
Biography:
Yuri Moskalenko was completed his PhD at age 29 years from Pavlov Medical University in Leningrad. He is specialist in brain circulatory physiology, focusing interest on the experimental study of pressure-volume relations of liquid media (blood, CSF) inside cranium by biophysical approach in Sechenov Institure Academy of Sciences R.F. He found relations, responsible for structural and functional stability of brain blood supply system at different conditions, including space flight. He has published 180 papers in reputed journals and 8 books, three of them published in USA and England. Awarded by the President of R.F. title â€Honor Scientist of Russian Federationâ€.
Abstract:
Slow volume fluctuations inside cranium have known since the middle of XIX Century. Although during passed time relations of their amplitude and frequency with some important physiological indices (pCO2, pO2 in brain tissue, functional activity of some brain structures, neurosurgical stages of brain activation) have been established, that these fluctuation couldn’t use in practice due to impossibility of their objective evaluation. Recently, spectrum analysis was applied for the study of intracranial slow volume fluctuations, recorded by coupling of noninvasive methods - multifrequency rheoencephalography and transcranial dopplerography with healthy volunteers and neurological patients. Spectrum analysis of 3 min fragments of recordings, provided by “Chart 5†software with quantitation 128 kHz, have shown, that frequency ranges 0.01 – 0,3 Hz include majority of spectral peaks, reflecting intracranial slow volume fluctuations and include also a few peaks, connected with of arterial pressure fluctuations and respiration. Comparison frequencies and amplitudes of spectral peaks, reflected intracranial volume fluctuations, show, that they are characterized by hemispheric asymmetry and by difference in surface and deep brain structures. Spectrograms, taken with patients, demonstrate increased number of peaks and decreased of their amplitude. After treatment, together, with disappearing of pathogenic symptoms, spectrograms are normalized. Received data indicate, that spectrum of intracranial slow volume fluctuations reflect the systems integration, (brain blood and CSF circulation as well skull mechanics) responsible for brain circulatory-metabolic supply. It is possibly to conclude, that spectral analysis of intracranial slow volume fluctuations, demonstrate quality of system integration in mechanism, which responsible for nutrition support of brain functioning.
Jiejun Wu
Janssen R&D US, USA
Title: The Development and Applications of two Novel Metabolomic Platforms for Drug Discovery Research
Biography:
Jiejun Wu obtained his B.S. in Biology from Peking University (Beijing, China), and Ph.D. in Biochemistry from The Ohio State University (Columbus, Ohio) under the direction of Prof. Alan G. Marshall. He has worked in both academic and pharmaceutical industry and is currently a Scientific Director in Discovery Sciences of Janssen R&D US, a pharmaceutial R&D division of Johnson & Johnson. He has published more than 40 papers in reputed scientific journals including Nature, PNAS, Anal. Chem., Biochemistry, J. Biol. Chem., and J. Org. Chem., etc..
Abstract:
Metabolomics is the "systematic study of the unique chemical fingerprints that specific cellular processes leave behind", which requires the identification and quantification of the endogenous small molecules in a biological sample and data mapping to metabolic pathways. A properly measured metabolic profile provides a comprehensive picture of the underlying physiology, and the profile may be more translatable across different species – in comparison to genetic, or transcriptional, or proteomic profiles. Applications of metabolomics in pharmaceutical R&D are many and rapidly expanding, such as disease characterization, drug efficacy and toxicity monitoring, and discovering individual metabolic characteristics. Together with GC/MS, NMR, and CE/MS, LC/MS is the most widely used technique in metabolomic studies. In order to focus on the desired metabolic pathways, targeted metabolomic panels (rather than global metabolomic profiling) should be employeed, for which very specific analytical protocols are usually required to achieve maximal sensitivity and throughput. Automated data processing and data analysis software tools also play a very important role in metabolomics. In this presentation, we will highlight two novel LC/MS-based targeted metabolomic platforms developed within Janssen R&D - a high sensitivity and high throughput eicosanoids platform and a high sensitivity oxysterols panel - with their application examples to various drug discovery projects.
Sejalkumari Keshavlal Patel
Ganpat University, India
Title: Development and validation of bioanalytical method for estimation of methoxsalen in human plasma
Biography:
I have completed my PhD at the age of 28 years from Ganpat University, Gujarat, India. I am assistant professor in Quality Assurance Department, Shree S. K. Patel College of Pharmaceutical Education and Research, Ganpat University, Gujarat, India since last 10 years. I have published more than 37 papers in reputed national and international journals.
Abstract:
In this study, an attempt was made to describe and validate a liquid chromatography coupled with tandem mass spectrometry method for the quantification of methoxsalen, an antipsoriatic agent in human EDTA K3 plasma according to the current bioanalytical guidelines. The internal standard used was methoxsalen D3. The separation was performed on a Symmetry, C18, 4.6X 150 mm, 5 µm column using a mobile phase of 2 mM ammonium acetate and methanol 15:85 (v/v) with a flow rate of 0.80 ml/min. The detection of methoxsalen and the internal standard was performed in multiple reactions monitoring (MRM) mode using LC/MS/MS Mass Spectrometer with electro spray ionization, operating in positive ion mode. The human plasma samples were extracted using liquid-liquid extraction with methyl tert-butyl ether. The method shows a good linearity (R2 > 0.98), precision and accuracy over the range of 0.1-100 ng/ml methoxsalen in plasma. The recovery was between 93.85 and 105.25%. The limit of quantification was 0.1 ng/ml. The analysis required about 3.2 minutes run with retention time of drug 2.2 minutes. The proposed method was selective, sensitive, accurate and precise enough to be successfully applied to bioequivalence study.
Vasile I. Furdui
Ontario Ministry of the Environment and Climate Change, Canada
Title: Ultra-trace level speciated isotope dilution measurement of Cr(VI) using ion chromatography tandem mass spectrometry in environmental waters
Biography:
Dr. Vasile Furdui is a senior scientist with the Ontario Ministry of the Environment and Climate Change and Adjunct Professor with Ryerson University (Toronto, Canada). In 2003 Vasile received his Ph.D. degree in analytical chemistry from University of Alberta, working with Prof. Jed Harrison on “Blood Sample Preparation Using Microfluidic Platformsâ€. His recent publications are focused on trace and ultra-trace analysis of various organic and inorganic species, including perfluorinated compounds, perchlorate and hexavalent chromium using liquid or ion chromatography coupled to tandem mass spectrometry. His top five publications have received over 580 citations.
Abstract:
The reliable analysis of highly toxic hexavalent chromium, Cr(VI), at ultra-trace levels remains challenging, given its easy conversion to non-toxic trivalent chromium. The new approach demonstrates a novel analytical method to quantify Cr(VI) at low ng/L concentration levels in environmental water samples by using speciated isotope dilution (SID) analysis and double-spiking with Cr(III) and Cr(VI) enriched for different isotopes. Ion chromatography tandem mass spectrometry (IC-MS/MS) was used for the analysis of Cr(VI) as HCrO4- → CrO3-. While following a classical linear multipoint calibration curve a method detection limit (MDL) of 7 ng/L Cr(VI) was achieved, the modified SID-MS method adapted from U.S. EPA 6800 allowed for the quantification of Cr(VI) with an MDL of 2 ng/L and provided results corrected for Cr(VI) loss occurred after sample collection. The adapted SID-MS approach proved to yield more accurate and precise results than the multipoint calibration method, allowed for compensation of Cr(VI) reduction during sample transportation and storage while eliminating the need for frequent external calibration. The SID approach permitted continuous sample analysis for several days without the need for recalibration. This new developed IC-MS/MS method represents an alternative to the routinely used inductively-coupled plasma (ICP) instrumentation, IC-ICP-MS, and offers several advantages over detection with ICP-MS for Cr(VI), such as the absence of polyatomic interferences of 52Cr formed in the ICP (36Arl6O, 40Ar12C, 35Cl16OH, and 37Cl14NH) that require the use of dynamic reaction/collision cells or high-resolution double-focusing sector field instruments.
K Srinivasa Rao
Anwarul Uloom College of pharmacy, India
Title: Identification and Characterization of New Drugs Using Mass Spectrometry
Biography:
Dr K Srinivasa Rao, working as Professor & HoD at Anwarul Uloom College of pharmacy, Hyderabad, India and obtained his bachelor’s and master’s degree from Andhra University, Visakhapatnam, Andhra Pradesh (India) and Ph.D and D.Sc from Berhampur University, Berhampur, Orissa (India) in Pharmaceutical Analysis. He has completed his` Doctor of Science (D.Sc) at the age 34 years. His area of interest is in phytopharmaceutical analysis & standardization and interpretation of IR, NMR, MS Spectra’s. He has several publications of national and international repute. He is a reviewer for many journals of national and international repute. He has guided 72 M.Pharm students, and a few students are pursuing Ph.D under his guidance.
Abstract:
Mass spectrometer can be used in 3 principal ways: firstly, to measure the molecular weights with very high accuracy; from these can be deduced exact molecular formulae. Secondly, to detect within a molecule the places at which it prefers to fragment; from this can be deduced the presence of functional groups within the molecule. And thirdly, as a method for identifying drug molecules by comparison of their mass spectra with libraries of digitized mass spectra of known compounds. Molecular ion gives highly useful information about the identity of the drug molecule. Fragmentation pattern gives further information about the structure of the drug molecule. All fragment ions are, however, not of equal significance to assign the structure to a compound. Intensity of the molecular ion peak in a mass spectrum depends on the type of the compound. Mass spectra cannot be interpreted if they contain any misinformation. Unfortunately, even referred journals and carefully edited collections of the standard spectra sometimes contains spectra that fail to meet this criterion. Some compounds e.g., alcohols may fail to give a visible molecular ion peak. Judging whether or not a mass spectrum is credible is sometimes the most critical step in its interpretation. Most EI mass spectrometers in use today lack sufficient resolving power to provide accurate mass measurement for the determination of elemental composition. However, the elemental composition of an ion can sometimes be determined from the ratios of the peak intensities of the isotope peaks for that ion to the intensity of the nominal mass peak. For eg., if the M+2 peak of the parent ion looks larger than the M+1 peak, the compound might contain S, Cl or Br. When there is a larger gap and a peak at 127, iodine may be present. The intensity of the M+1 peak can be used to know the number of carbons well as nitrogen atoms. In the absence of nitrogen, the maximum number of carbaon atoms can be calculated by dividing the relative intensity of the M+1 peak by 1.1. In case nitrogen is present its contribution to the M+1 peak will amount of 0.4 X number of nitrogen atoms. This quantity must be subtracted from the measured relative intensity of the M+1 peak to know the number of carbon atoms. In this talk we will address, how to identify different drug molecules using mass spectrophomter with relevant examples.
Clemens Schwarzinger
Johannes Kepler University Linz, Austria
Title: Sample preparation in MALDI ToF Mass Spectrometry
Biography:
Clemens Schwarzinger has completed his PhD at in 2002 from the Johannes Kepler University Linz (Austria). After industrial PostDocs and being guest professor at the Petroleum Institute in Abu Dhabi (UAE) he is associate professor at the Institute of Chemical Technology of Organic Materials at the Johannes Kepler University Linz. The main research interests are polymer mass spectrometry, analytical pyrolysis and the development of new polymer systems, but also the use of mass spectrometry in gemology. He has published more than 50 papers and gave more than 120 presentations at various conferences.
Abstract:
Polymer mass spectrometry, especially MALDI-ToF MS, has gained a lot of attention in the last years because of the high amount of information that can be gained, such as molar mass distribution, repeat units, end groups, etc. But care must be taken when it comes to sample preparation, especially when the simple and fast “dried droplet†technique is used. As most people have already experienced dried droplet preparation tends to from rings of higher concentration at the outer rim, the so called “coffee ringsâ€, which results in inhomogeneous distribution of the compounds and therefore questionable results. We will show that several rather simples steps can be used to circumvent this phenomenon and how to produce reliable high quality data using dried droplets, as there are the use of ionic liquids as matrices or higher matrix concentration. The results were monitored with imaging techniques such as FTIR or mass spectrometric imaging to understand the processes necessary for a perfect sample preparation. When it comes to copolymers things are getting even worse. In this case it is mostly necessary to separate the polymer into fractions either by precipitation or by SEC. We have found that a modified Electrospray interface coupled to the SEC is a very efficient and elegant way yielding best results in terms of polymer separation and MALDI sample preparation.
Tatsuya URABE
Nishina center for Accelerator-Based Science, RIKEN, Saitama, Japan
Title: Introduction of liquid samples into high-vacuum plasma ion source for low-cost quantification of inorganic elements
Biography:
Tatsuya Urabe has completed his PhD in 2010 from Tokyo University of Marine Science and Technology and worked as a postdoctoral researcher of Japan Society for the Promotion of Science (JSPS) until 2011. Now he is working in an accelerator facility at RIKEN and developing a new mass spectrometer based on accelerator science.
Abstract:
We have been developing a mass spectrometer based on plasma ion source, ECRIS (Electron Cyclotron Resonance Ion Source). Although ECRIS is known as a large-sized ion source in accelerator facility, we succeeded to miniaturize and mount it on a portable mass spectrometer (called “miniECRIS-MSâ€) for detecting gas compounds1). The miniECRIS-MS contains a permanent magnet for an ion source, ion optical system, quadrupole mass analyzer and so on. In this poster session, we will present our ongoing project for quantifying elements in liquid samples. Comparing gas samples, liquid sample introduction into high vacuum plasma (i.e. ECRIS) is troublesome. However, it can offer great advantages over present analytical methods such as ICP-MS in terms of stability, sensitivity, freedam from polyatomic (spectral) interence, initial/running costs, and portability. Details of sample introduction system are as follows. Electrospray is used as a splaying unit of liquid samples at first stage.The solutions containing the analyte of interest are electrosprayed and gradually desolvated with the aid of heated assist gas at atmospheric pressure. Then, sample aerosol was introduced into ECRIS through differential pumping systems. Since analytes of interest are charged by electrospray (as relatively large clusters), they can be electrically extracted by ion lenses such as ion funnel in the pumping system. And they are finally reached at ECRIS and ionized (decomposed) into monvalent atomic ions for quantitative purpose. By this system some solvents (water and methanol) and inorganic elements (Na) were successfully observed, and the potential of miniECRIS-MS will be discussed in the session.
Weiwen Chen
Guangzhou University of Chinese Medicine, China
Title: UPLC-Q-TOF/MS-based screening and identification of the main alkaloids and their metabolites in rat plasma,urine and feces after oral administration of Zanthoxylum nitidum decoction
Biography:
Professor Weiwen Chen obtained his PhD in cell biology in 1988 from the Xavier-Bichat Medical School, Université Paris Diderot-Paris 7, France. He is currently the Vice Principal of Guangzhou University of Chinese Medicine. His research interests focus on curing gastric-intestinal diseases using integrated traditional Chinese and western medicine, as well as on novel drugs R & D. He has published more than 100 papers in reputed journals.
Abstract:
Zanthoxylum nitidum (Roxb.) DC (Rutaceae), called Liangmianzhen in China, is used traditionally for several medicinal purposes. Its pharmacological effect have been primarily attributed to the presence of a number of alkaloids. However, due to lack of metabolism research, its main bioactive alkaloids in vivo is still unknown. A systematic method based on ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS) technique combined with MetabolitePilot software was developed to speculate the metabolites and excretion profiles of the main alkaloids in Zanthoxylum nitidum decoction in rats plasma, urine and feces samples after oral administration of the decoction. As a result, 6 parent components and a total of 18 metabolites of 3 main alkaloids including magnoflorine (MF), α-allocryptopine (AC) and skimmianine (SA) were tentatively detected in vivo. All metabolites were detected including hydroxylated, demethylated, ketonization products and their sulfate and glucuronide conjugates. 11 metabolites were from the rat plasma, 14 from the urine and 8 from the feces. Among them, metabolites of AC and SA were reported firstly. In conclusion, the research provided useful information for further study of the pharmacology and mechanism of action of Zanthoxylum nitidum decoction in vivo.
Igor Spivak-Lavrov
Aktobe Regional State University, Kazakhstan
Title: Time-of-flight mass analyzers based on a wedge-shaped electrostatic mirror with 2d field
Biography:
Igor Spivak-Lavrov holds a degree – Doctor of Physical and Mathematical Sciences (USSR). He defended his doctoral thesis on the problems of optics of charged particles at the age of 54 years at the St. Petersburg Technical University. He is a professor in the Department of Theoretical Physics of Aktobe Regional State University (Kazakhstan). He has published more than 200 papers in reputed journals.
Abstract:
The paper deals with the wedge-shaped mirror with two-dimensional (2D) field as an orthogonal ion accelerator and a TOF mass analyzer. The inhomogeneous electric field in the mirror created without the use of meshes and blinds by changing the potentials on the boundary electrodes of mirror. Analytical expressions for the electrostatic potential of mirror field obtained. The ions are extracted from the ion source by impulse activation of the accelerating electric field. Since the ions are extracted from various points of the source, even ions with identical masses acquire different velocities during acceleration. As a result, the “rear†ions of the packet catch up with the “front†ions, and packets of ions with identical masses compress in the direction of movement. By placing the detector in a plane with the greatest compression of ion packets, one creates a time-of-flight mass spectrometer. Efficient space-time focusing allows a small mass spectrometer to achieve high resolution and sensitivity. The paper also suggests and calculates new schematics of multi-stage and multi-reflection TOF mass analyzers with wedge-shaped mirrors with 2D field.
Pankaj Pathak
All India Institute of Medical Sciences (AIIMS), India
Title: IDH1 mutations in gliomas: First series from a tertiary care centre in India with comprehensive review of literature
Biography:
His research interests span from nanoparticle detection to analytical method development involving plasma spectrometry. He published more than 50 papers, served as guest editor and reviewer for major analytical and environmental chemistry journals.
Abstract:
Mutations of the gene encoding isocitrate dehydrogenase (IDH) have been shown in a significant proportion of diffuse gliomas. These mutations are specific to gliomas and their utility for diagnosis and prognostication of these tumors is being proclaimed.IDH1 mutations in codon 132 were observed in 46% cases. The frequency was 68.8% in grade II, 85.7% in grade III and 12.8% in GBMs. R132H mutation was most frequent (84.8%). Overall frequency of these mutations was relatively higher in oligodendroglial tumours as compared to astrocytic phenotype (66.7% versus 38.4%; p = 0.06). Primary GBMs showed IDH1 mutation in only 4.4% cases. In contrast, 66.7% of secondary GBMs harboured this alteration. Patients with IDH1 mutations were significantly younger as compared to those without mutation (p = 0.001). There was a significant correlation between IDH1 mutation and TP53 mutation (p = 0.004). Although IDH1 mutation showed a positive correlation with 1p/19q deletion, the association was not statistically significant (p = 0.653). There was no correlation with EGFR amplification or PTEN deletion.
Pham Thi Tam Ngoc
University of Natural Ressource and Environnement, Vietnam
Title: Thermodynamic assessment of Cs-Te system
Biography:
Pham Thi Tam Ngoc has studied in Engineer School of Chemistry of Rennes, France in 2005. She held an engineer degree in 2010 in Pau, France. Since 2010, she has worked for the project of “Haracterization and modelling of the thermodynamic behavior of SFR fuel under iradiationâ€- the co-operation research project between Aix-Marseille University and CEA Research Center. Accomplished the PhD. During this period of time, she had prestigious opportunity to work with high-profile experts in employing CALPHAD methology. At the age of 28 she has then worked as a Lecturer at the University of Natural Ressource and Environment, Ho Chi Minh city, Vietnam, 2015.
Abstract:
The operating conditions of mixed oxide fuels (MOX) in Sodium cooled Fast Reactor (SFR) are very severe combining high temperature, high linear rating and high temperature gradient. In the nominal operating conditions of a SFR, the linear power is around 2 kW/cm3 and the temperature varies from 2500K down to 800K between the center and the periphery of the oxide fuel pellets. Due to those conditions, the volatile Fission Products (FP) like cesium and tellurium generated in the central region of the fuel pellet migrate outward through the radial cracks of the fuel matrix. At high burn up, a mixture of compounds of FP is formed in the fuel-cladding gap. This layer of FP compounds located between the external surface of the fuel pellet and the inner cladding surface is called in french the Joint Oxyde Gaine (JOG). The knowledge of phase equilibria and thermodynamic properties of the Cs–Te system is thus crucial for understanding and modeling the diffusion processes during the formation of the JOG. In this work, we present the review of phase diagram, crystallographic data and thermodynamic data of the Cs–Te binary system. The thermodynamic modeling of this system is also performed with the aid of the Thermo-Calc software. The thermodynamic descriptions derived in this work are based on the databases of Scientific Group Thermodata European (SGTE) and TBASE (ECN, Petten, Netherland) for the pure elements and the gaseous species. The compound formation and liquid mixing Gibbs energy ex- pressions are obtained by a least square optimization procedure. Comparisons between calculated and available experiments results are presented. A satisfactory agreement is achieved.
Biography:
Dr. Biljana Škrbić is full professor at the Faculty of Technology, University of Novi Sad (FoTNS) with significant experience in coordination and leadership of research projects at national and international levels; founder and head of the Centre of Excellence in Food Safety and Emerging Risks (2009-) and Laboratory for Chemical Contaminants and Sustainable Development (2015-) at FoTNS; co-editor of Central European Journal of Occupational and Environmental Medicine; vice-editor of Environmental Processes (Springer). She delivered 23 lectures as invited speaker (int. conferences) and published 107 articles in the leading international journals from SCI list with impact factors of which 9 articles are dedicated to UHPLC-MS/MS.
Abstract:
As a crude extract method followed by ultra-high performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) is a technique that has been frequently used as a routine analytical technique for mycotoxins in wheayt and corn, which applicability has been also demonstrated successfully on other matrices by the team, the intention of this presentation was to further provide the evidence on the method applicability for mycotoxin analysis of rice samples, for UHPLC–MS/MS represents the most efficient and reliable analytical technique for high efficiency isolation, unequivocal identification and accurate quantification. Thus, in present investigation, a reliable UHPLC–MS/MS method was developed and validated for simultaneous determination of twelve mycotoxins (aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, ochratoxin A, zearalenone, T-2 toxin, HT-2 toxin deoxynivalenol, citrinin, fumonisin B1 and fumonisin B2) in rice. One-step extraction using solvent mixtures of acetonitrile/water/acetic acid (79:20:1, v/v/v) without any clean-up (i.e. crude extract method) was employed for the mycotoxins extraction from rice samples. Using matrix-matched calibration it has been demonstrated that the limits of quantification were below the relevant maximum contents established by the EU regulation in rice or other cereals. The recoveries of the developed method were satisfactory between 96%–126% with relative standard deviation lower than 12%. The validated method was applied on real samples. The obtained results are used to estimate the exposure of the Serbian population to mycotoxins. To the best of our knowledge, this is the first report on occurrence of the studied mycotoxins in rice commercially available in Serbia.
MD Ahsan Habib
The University of Tokyo, Japan
Title: Development of new ion source/new desorption method for mass spectrometry
Biography:
Md. Ahsan Habib has completed his PhD in Analytical Chemistry at the age of 36 years from Saga University, Japan. Currently Dr. Habib working as a JSPS fellow at the University of Tokyo and Professor, Department of Chemistry, University of Dhaka, Bangladesh. Dr. Habib has also completed 06 years 04 post-doctoral research at Saga, Chiba, University of Yamanashi and Yokohama National University in various fields of Chemistry/Science. He has published more than 30 papers in reputed journals and presented in about 30 in national/international seminars/conferences.
Abstract:
Detection and quantification of illicit compounds at trace level is very much important for public health, security and safety. Mass spectrometry (MS) has already been demonstrated its versatility for detection and quantification of a wide range of compounds at trace level because MS can provide molecular level information of the target compounds. In MS, there are many ionization sources. So far, none of the ion source can ionize efficiently of a wide range of compounds. For example, electrospray ionization (ESI) and nano-ESI have been widely using for bio-molecules. Nano-ESI has shown better resolution than ESI but it has clogging problem. To overcome such a problem, several attempts have been taken to develop new ionization source, for example, MALDI, probe-ESI (PESI) etc. Recently we have developed new atmospheric pressure chemical ionization (APCI) using alternating current (ac) instead of dc and found as a soft ionization source for explosives detection (Habib et al., RCMS). Moreover, hollow cathode discharge (HCD) ion source has been fabricated for detection of explosives at trace level (Habib et al., RCMS). A desorption method has also been developed using an ultra-cutter to desorb highly non-volatile illicit compounds and ionized dielectric discharge (DBD) ion source, and found better limit of detection (LOD) (Habib et al., ASMS). A vacuum glow discharge ionization (vacuum-GDI) source has been fabricated for the compounds those give negative ions. A further attempt has also been taken to fabricate a hybrid ionization source for polar/non-polar, volatile/non-volatile compounds.
Biography:
Zikri Arslan received his PhD in 2000 from the University of Massachusetts at Amherst in analytical chemistry with an emphasis on applied plasma source mass spectrometry. He was a National Research Council (NRC) post-doctoral scholar between 2000 and 2002; worked at NOAA/NEFSC Howard Marine Sciences Laboratory in Sandy Hook, NJ. He also worked as research scientist (2002-2003) at the University of Maryland, Chesapeake Biological Laboratory on otolith micromilling protocols for identification of bluefin tuna stocks. His research interests span from nanoparticle detection to analytical method development involving plasma spectrometry. He published more than 50 papers, served as guest editor and reviewer for major analytical and environmental chemistry journals.
Abstract:
A chemical vapor generation (CVG) method is developed for determination of cadmium (Cd) by ICP-MS. Titanium (III) and titanium (IV) were utilized for the first time as novel additives to enhance generation of volatile Cd species, and their synergistic effects were investigated for thiourea, L-cysteine and potassium cyanide (KCN) with different mineral acids. Both Ti(III) and Ti(IV) provided the highest enhancement with KCN. The improvement with thiourea was marginal (ca. 2-fold), while L-cysteine boosted signals slightly only with Ti(III) in H2SO4. Optimum CVG conditions were 4% HCl + 0.03 M Ti(III) + 0.16 M KCN and 2% HNO3 + 0.03 M Ti(IV) + 0.16 M KCN. A 3% (m/v) NaBH4 solution was adequate for successful vapor generation. Under these conditions, sensitivity was enhanced 40-fold with Ti(III) and 35-fold with Ti(IV), that are the highest enhancement factors achieved so far in Cd vapor generation. Detection limits (3s) were 3.2 and 6.4 ng L-1 for 111Cd isotope using Ti(III) and Ti(IV), respectively. Evidence indicated that Ti(III) and Ti(IV) enhanced Cd vapor generation catalytically. Effects of transition metal ions, including Co(II), Cr(III), Cu(II), Fe(III), Mn(II), Ni(II) and Zn(II) were not significant up to 1.0 µg mL-1. Among hydride forming elements, Bi, Pb, Sb and Sn depressed signals above 0.1 µg mL-1. No interferences were observed from As(III) and Se(IV). The method was validated with determination of Cd by CVG-ICP-MS in certified reference materials, including Nearshore seawater (CASS-4), Bone ash (SRM 1400), Dogfish liver (DOLT-4), Mussel tissue (SRM 2976) and Domestic Sludge (SRM 2781).
Brigitte R. S. Pecequilo
Cidade Universitária, Brazil
Title: Self-attenuation factors in gamma-ray spectrometry
Biography:
Brigitte Pecequilo completed her PhD in 1977 from the University of São Paulo, Brazil, with a major in Nuclear Technology. She is presently Senior Research Scientist with the Environmental Radiation Laboratory at the Instituto de Pesquisas Energéticas e Nucleares, São Paulo, Brazil. Her investigations include radioecology, environmental radioprotection and radiation detection measurements. She has published more than 25 papers in reputable journals and oriented several students for Master and PhD degrees. Since 1978 she is also a partial-time professor of Advanced Topics in Nuclear Measurements at the University of São Paulo.
Abstract:
Gamma-ray spectrometry with high-resolution semiconductor detectors is currently the most widely used analytical technique for qualitative and quantitative determination of gamma-ray emmitters radionuclides. Quantification of elements relies on the correct analysys of the spectra, depending strongly on the efficiency calibration of the measurement apparatus, most often performed with aqueous standard multi-radionuclides solutions. Soever, lower energy gamma rays have less penetrating ability and tend to interact more readily with matter, so, when the efficiency calibration curve is obtained with an aqueous standard multi-radionuclides solution, a self- attenuation correction is required if samples present densities higher than the water ones. Samples such sand, soils and sediments have apparent typical densities varying from 1.2 g.cm-³ to 2.9 g.cm-³, so a correction of the efficiency curve is necessary. Self-attenuation factors are determined by Cutshall transmission technique, measuring the transmission of gamma rays through both the solid sample and an ultrapure water sample in the same geometry, using punctual sources, with gamma transitions in the range of interest. All samples are measured in the same geometry with high-resolution HPGe detectors. Self-attenuation correction factors are experimentally determined for every different sample density. A study of two hundred samples of sands and soils, allow to conclude that, to obtain more reliable analyses, one should use correction factors for the self attenuation behavior, especially in the lower part of the energy spectrum. Also, as attenuation, apart from density, strongly depends on the sample chemical composition, correction factors must be determined for each studied sample.
Mikhail Bolshov
Moscow State University, Russia
Title: Mass spectrometry with inductively coupled plasma as a technique for analysis of complex matrix.
Biography:
Dr. Mikhail Bolshov has completed PhD in 1973 from the Department of Physics, Moscow State University. 1993 has completed Dr.Sci. degree from Institute for Spectroscopy, Russian Academy of Sciences. Currently the head of the laboratory in the Institute. Since 2006 take a half time position of the Professor at the Department of Chemistry, Moscow State University. 1996 - invited Professor at the University of Florida (Gainesville), 1998-2000 the head of the laboratory in ISAS (Dortmund, Germany). Author and co-author of more than 120 publicationsand in WoS journals. Since 1988 the member of Editorial Advisory Board of Spectrochim. Acta B.
Abstract:
Mass spectrometry with inductively coupled plasma (ICP MS) takes the leading position among the instrumental techniques for elemental analysis. Main advantages of ICP-MS are: extremely high sensitivity (limits of detection down to ppt), multielement capability (detection of the elements from Li to U in one cycle from sample volume about 1 mL), high throughput, user friendly operation (not high qualification of an operator), gradually decreasing price. However as all instrumental techniques, ICP-MS suffers from spectral interferences and matrix effects – the dependence of analytical signal on the composition of a sample. These effects can strongly affect the accuracy of the analysis. Different types of interferences exhibit serious role in the analysis of the samples with complex composition e.g. biological samples (human liquids, bones, hairs), pharmaceuticals, strongly acidified resulting liquids after a solid sample digestion and so on. The strategy of accounting for or, et best, eliminating these effects depends on the types of the ICP-MS instrument. Basically two types of instruments are now widely used – quadrupole ICP MS (QMS) and Sector Field MS (SFMS). The first has low mass resolution (not better than 600), but is much cheaper. The second has resolution up to 7000, but is about 3-4 times more expensive. Evidently, the QMS instruments occupy larger part of the market. The problems of interferences are much stronger pronounced for the QMS instruments. The results obtained in my group with a simple QMS will be presented in the talk. Different strategies of the accounting and elimination of the interferences will be exemplified on the developed strategy of the analysis of bio-samples and strongly acidic liquids after rock sample digestion. In case of bio-sample analysis two techniques of a sample pretreatment were compared - 1) acid mineralization (in open or closed vessels or MW digestion), and 2) dilution by the mixture of reagents. It was found that combination of instrumental parameters optimization and use of internal standard (IS) provides adequate accuracy of the determination in blood and hair samples for crucial elements - Al, Cr, Mn, Co, Cu, Zn, As, Se, Cd, Pb. Not trivial was the finding of the analytical strategy, which allows using one and any IS independent on its mass or ionization potential. Another approach was developed for analysis of acidic liquids. The pretreatment procedure and measurement strategy will be discussed in the talk. In the end a brief overview of recent improvements of QMS instruments will be presented. These results are based on my lectures in MSU.
Grazieli Simões
Universidade Federal do Rio de Janeiro, Brazil
Title: The use of mass spectrometry and spectroscopic techniques to study sulfur containing biomolecules
Biography:
Grazieli Simões has completed her PhD at Universidade Federal do Rio de Janeiro, Brazil, in 2013 with a one-year fellowship grant at the french synchrotron light source SOLEIL. She was a researcher and a teacher at Université Paris-Sud in France from 2014 to 2016. She has recently obtained a permanent position at Universidade Federal do Rio de Janeiro in Brazil. Her last publications involve the use of different types of mass spectrometry techniques, using synchrotron radiation and electron impact techniques.
Abstract:
In recent years, there has been a large interest in the study of biological systems, namely, amino acids, peptides, and proteins, using synchrotron-based spectroscopic techniques, such as near-edge X-ray absorption fine structure (NEXAFS) or related X-ray photoelectron and X-ray emission spectroscopies. Most of the X-ray spectroscopic investigations of biologically relevant molecules, such as amino acids and their polymers, have been performed on thin organic films and liquids. The association of mass spectrometry and spectroscopic techniques has allowed for the investigation of the effects of radiation damage in sulfur containing molecules. We have performed a NEXAFS (S1s) and mass spectrometry study of solid samples of cysteine, cystine and insulin irradiated with 0.8 keV electrons. The measured mass spectra point out to processes of desulfurization, deamination, decarbonylation and decarboxylation in the irradiated biomolecules. In another study, inner-shell measurements of insulin were performed by coupling a linear ion trap mass spectrometer, equipped with an ESI source at the french synchrotron radiation facility SOLEIL. The electrosprayed insulin ions were injected, mass selected, stored in the trap, and irradiated during a well-defined period. The near-edge X-ray ion yield spectra of the 6+ charge state insulin precursor were recorded as a function of the photon energy, in the vicinity of the C1s edge.
Magnus S. Magnusson
University of Iceland, Iceland
Title: Similarity of Hierarchical Structured Clustering in Human and Neuronal interactions and on DNA: Structural and Functional analogies.
Biography:
Magnus S Magnusson, Research Professor. PhD in 1983, University of Copenhagen. Author of the T-pattern model and the corresponding detection algorithms in THEME. He has focused on real-time organization of behavior, co-directed DNA analysis, numerous papers and invited talks at numerous conferences and universities in Europe, USA and Japan. Deputy Director 1983-1988, Anthropology Laboratory, Museum of Natural History, Paris. Repeatedly invited temporary Professor at the University of Paris. Since 1991, Founder and Director of the Human Behavior Laboratory, University of Iceland. Since 1995, he is in collaboration between 24 universities based on “Magnusson’s analytical model†initiated at the Sorbonne, Paris.
Abstract:
Hierarchical structured clustering (HSC) seems characteristic of the structure of the universe balancing a small number of forces, some pulling others pushing apart, the self-similar fractal distribution of matter in the universe thus reflecting HST rather than just dispersion or clumping. HSC also characterizes a proposed pattern type, called T-pattern, detected in the temporal organization of many kinds of verbal and non-verbal human, animal and neuronal behavior and interactions and is also characteristic of the structure of DNA. Functional analogies seem to exist between the occurrence of T-patterns in “cell city†and in human cities. Structural self-similarity over many levels of biological organization suggests the possibility of a unified (mathematical, bioinformatics and system biological) approach. The T-pattern is described as a repeated hierarchical and self-similar tree structure based on a single non-terminal relation, called a critical interval (CI) relationship. The instances of a T-pattern may be seen as repeated statistical pseudo-fractal objects characterized by statistically significant translation symmetry. THEMEtm (by M.S. Magnusson, ©PatternVision Ltd) is special purpose T-pattern detection and analysis PC software using a special CI detection algorithm combined with an evolution algorithm, presented here together with results from the analysis of behavior and interactions. From the relatively slow time scale of human and animal interactions to the much faster interactions within populations of neurons in living rat brains. Analogies are discussed between T-pattern structure and functions in the „cities of proteins“ (cell city) and human cities, especially regarding specialization and the particular case of religious behavior.
Biography:
Dr. Simone König completed her PhD in Physical Chemistry at the University of Leipzig and time-out for motherhood, Simone König has done postdoctoral studies with one of the pioneers in biomedical mass spectrometry, Henry M. Fales, at the National Institutes of Health in Bethesda, MD. For over a decade, she heads the Proteomic Facility of the Interdisciplinary Center for Clinical Research at the Medical Faculty of the University of Münster, Germany. She has published more than 100 peer-reviewed papers and written and edited books. She serves as a reviewer for national and international funding organisations as well as journals and she organizes the Annual Münster Conference on Biomolecule Analysis.
Abstract:
A major problem for the acceptance of two-dimensional (2D) protein gel electrophoresis in proteome expression analysis has been the little comparability of gel images among different laboratories or even within the same working group. Now, the generation of a reference grid of marker proteins in parallel to the separation of the analyte allows the correction of protein coordinates both in x (protein isoelectric point)- and y (molecular weight)-direction. This is achieved by use of multiple fluorescent dyes for standards and samples on the same gel. A net of ~100 marker nodes spans the gel and provides reference points for triangulation. The deviation from mean is improved by an order of magnitude. The technology termed Comparative 2D Fluorescence Gel Electrophoresis (CoFGE) can be carried out in both vertical and horizontal electrophoresis instrumentation with the latter being much more comfortable. With CoFGE, archiving of coordinates of proteins identified by mass spectrometry in databases has become reality and a Repository of Gel-Separated Proteins (ReGeSeP) was generated for storing and searching protein IDs, coordinates and proteome gel images. The technology was protected (Patent application EP11167383.6, May 25, 2011), licensed, and awarded with the WWU Transfer Prize 2013/2014.
Biography:
Dr. Brigitte Simons is a market development specialist at SCIEX, specializing in metabolomics using accurate mass time-of-flight mass spectrometry solutions. Prior to working at SCIEX, Brigitte received her Ph.D. in Chemical Biology at the University of Ottawa. She then completed two research post-doctoral fellowships at the Centre for Biologics Research at Health Canada and the National Institute of Heart Lung and Blood in Bethesda MD. Brigitte also spearheads an academic partnerships program for our North American business – which listeners can browse sciex.com for more information or contact Brigitte for more information.
Abstract:
This presentation will address technologies that directly address clinical research experimentation of lipids in tissue extracts, keeping sampling and data processing throughput in mind. The human lipidome contains >100,000 different molecular species found within a small mass range; consequently, isobaric overlap makes unambiguous identification and quantitation of lipid species difficult. Herein, methods that utilize high sensitivity MS/MS techniques using a high sensitivity triple quadrupole linked LIT mass spectrometer to isolate lipid classes for identification of molecular composition and quantitation have been investigated. Furthermore, QqQ platforms can now be coupled to differential ion mobility (DMS) devices and have shown to resolve phospholipid sub-classes, triglycerides, and strikingly, the sphingomyelins (SM) were resolved from PC molecular species. The latter observation is significant considering these two classes cannot be resolved using triple quadrupole strategies alone, and their masses overlap significantly. Alterations in hepatic phospholipid composition, especially the amount of phosphatidylcholine (PC) and the ratio of PC to phosphatidylanolamine (PE) might contribute to the development non-alcoholic fatty liver disease (NAFLD). Aim was to compare PC/PE ratio in patients with NAFLD or chronic Hepatitis C (CHC) to healthy controls. This was a cross-sectional study. Liver samples were obtained from healthy controls (HC), patients with simple steatosis (SS), and steatohepatitis (NASH). Lipids were extracted from liver tissue and the total PC and PE lipid detected was quantified by tandem mass spectrometry using multiple precursor ion scanning [normalized by internal standards]. Hepatic PC/PE ratio was lower in all 3 patient groups, mainly due to a reduction in PC, which was significant in SS and NASH compared to controls. This study confirms previous animal data on PC/PE in NAFLD and represents a high throughput lipidomics platform’s utility to clinical outputs.