International Summit on Current Trends in Mass Spectrometry July 13-15, 2015 New Orleans, USA

Biography:

Olivier Laprévote has completed his PhD at the Paris-South University and joined the CNRS Institute of Natural Products Chemistry in 1989. He is now full Professor of Toxicology, Director of the Research Unit CNRS 8638 of the University Paris Descartes, and Director of the Biological Toxicology Department of the Paris Hospitals. He has published 180 papers in reputed journals and is board member of international journals (J. Mass Spectrom., Eur. J. Mass Spectrom, Anal. Bioanal. Chem.). He is past-President of the French Mass Spectrometry Society as well as the French representative at the International Mass Spectrometry Foundation

Abstract:

Mass Spectrometry Imaging (MSI) is now a mature analytical technique since its appearance more than fifteen years ago. Its ability to localize a variety of biomolecules on biological section surfaces and its promises in the medical field was underlined from the beginning of the IMS development. However, many years after, it seems that MS imaging in its two main instrumental modes i.e. Matrix-Assisted Laser Desorption-Ionization (MALDI) and Secondary-Ion Mass Spectrometry (SIMS) are still in their early developments for the discovery of biomarkers of diseases. Some recent results have demonstrated that MSI techniques can be used to image low mass endogenous metabolites in addition to proteins in biological tissue sections. By its ability to locate on histologically well-defined areas of tissues previously intractable classes of molecules, MSI offers enormous clinical potential. The reasons for such a delay between the early expectations for the MSI development and the actual applications of the technique for localized proteomics and/or metabolomics is due to many factors, including sample handling, instrumentation or even bio-informatics limitations. Some improvements have led to promising results in localized proteomics and metabolomics and the main goal is now to combine the MS imaging data with biochemical, signalling and metabolic pathways together with histological features. All these various aspects will be discussed in the lecture, in order to highlight the present state and the future trends of MS imaging combined with omics studies..

Biography:

Vijayabhaskar V, Ph.D., GVK BIO: Heading the department of ADME-Tox (Drug Metabolism & Pharmacokinetics) at GVK BIO, based in Hyderabad, India. He has received his Ph.D. in Pharmaceutical Sciences from the Department of Pharmacy at JJT University. Research during Vijay’s professional career was focused on understanding and troubleshooting technical issues in PK, In vitro ADME and Bioanalysis. A gold medallist in Master’s Degree of Pharmacy, he possesses an industrial experience of close to 10 years in different disciplines of DMPK. His current research interests include metabolite identification and high throughput bioanalysis.

Abstract:

Formulation excipients are used at high concentrations in preclinical formulations to solubilise NCEs of varying lipophilicities. The good (Measuring plasma concentrations of formulation excipient acts as quality control check for in vivo PK studies), the bad (Matrix effects arising from formulation excipients cause differential ionisation between calibration standards and study samples, producing false concentration levels) and the ugly (altered pharmacokinetic parameters resulting in false positive and false negatives) aspects of using formulation excipients in preclinical formulations will be discussed in detail. Bioanalytical approaches to identify and nullify the matrix effects will also be presented.

Biography:

Gilmare Antônia da Silva has completed his PhD at the age of 30 years from State University of Campinas (UNICAMP/BRAZIL). She is adjunct professor at Federal University of Ouro Preto/Brazil. She has published 15 papers in reputed journals and participated in the filing of a patent. She received two awards for two of the more than 70 works presented at scientific meetings. She has more than 25 academic supervisions including four mastering students.

Abstract:

Contaminants of emerging concern are organic compounds used in large quantities by the society for various purposes. They have shown biological activity at low concentrations and the difficulty to detect and quantify them in the environment stimulates the development of appropriate analytical methods. In this work, a chemometric approach to positive and negative ESI optimization for the simultaneous determination of contaminants of emerging concern in water samples by LC-IT-TOF-HRMS was applied. Three types of phase modifiers were used: formic acid, ammonium hydroxide and formic acid/ammonium formate. The effects of the operational parameters mobile phase modifier concentrations, mobile phase flow rate, heating block temperature and drying gas flow rate were evaluated by the 24−1 fractional factorial and Doehlert experimental designs. Factorial design indicated that ammonium hydroxide was more efficient compared to the other evaluated modifiers (higher ion intensities). Doehlert design allowed finding a region indicative of the optimum experimental conditions for most analytes. The best experimental condition observed was 3.5 mM ammonium hydroxide concentration; 0.0917 mL/min of mobile phase; 300 ºC heating block temperature; and drying gas at 200 kPa. These optimized parameters resulted in decreased detection limits of the method. The optimized method was applied to the evaluation of water samples coming from the Rio Doce basin - Minas Gerais/Brazil utilizing principal component analysis and Kohonen neural network. In this way, the use of chemometric approaches showed to be a promising way to optimize the simultaneous determination of 21 contaminants of emerging concern in water by LC-IT-TOF-HRMS using ESI. Biography

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 120 scientific papers and a score of patents.

Abstract:

Forensic engineering of advanced polymer materials (FEAPM) deals with the evaluation of the relationships between their structure, properties and behavior before, during and after practical applications. It is of particular importance in the case of nanomaterials and their nano-safety, biodegradable polymer materials as well as nanocomposites. The ex-ante investigations as well as the ex-post studies are needed in the area of FEAPM in order to increase efficiency and to define and minimize potential failure of novel polymer products before and after specific applications. Due to the wide spectrum of their potential applications e.g.: in medicine, in the field of compostable polymer packages (especially of long-shelf life products such as cosmetics or household chemicals) as well as in agrichemical formulations the FEAPM can provide basic knowledge and a valuable service by increasing understanding and helping prevent future problems. Such approach helps to design novel biodegradable polymer materials and to avoid failures of the commercial products manufactured from them. It also opens a wide opportunities for FEAPM mass spectrometry. Contemporary report on the MS applications for forensic engineering of natural aliphatic (co)polyesters (PHA) and their synthetic analogues, formed via anionic ring-opening polymerization of β-substituted β-lactones, will be presented. Special emphasis will be given to the results of ESI-MS ecotoxicological studies of polyester blends containing atactic poly(3-hydroxybutyrate). Furthermore, the use of environmentally friendly polymers as packaging materials for long shelf-life applications is the new trend for production. Thus, the results of the ESI-MSn investigations on PLA materials for cosmetic packages will be presented based on the studies of both the eroded polymer and its degradation products formed in paraffin and selected protic media.

Biography:

Tanumihardjo manages a research team studying vitamin A and carotenoid metabolism, serves as director of the Undergraduate Certificate in Global Health, and teaches at the undergraduate and graduate levels including international field experiences. She is on the Executive Board for the UW Global Health Institute. Tanumihardjo has >150 publications and chapters. She has presented at >200 domestic and international meetings. She has served as a reviewer for many journals. Awards: WHO’s Expert Advisory Panel, G. Malcolm Trout visiting scholar at Michigan State University, Ruth Pike Lectureship at Pennsylvania State University, Alex Malaspina ILSI Future Leader, and Dannon Creative Leadership Institute.

Abstract:

Vitamin A is an essential nutrient important for vision, reproduction, immune function, and cellular differentiation. Vitamin A status assessment is not straightforward. Serum retinol (vitamin A) concentrations are homeostatically controlled over a broad range of liver vitamin A concentrations, which are how vitamin A status is defined. Moreover, infection and inflammation decrease circulating retinol concentrations further confounding assessment. Retinol isotope dilution (RID) methods using a variety of mass spectrometric techniques are gaining momentum on the continent of Africa. A recent study in Zambia used 13C-labelled retinyl acetate to assess total body vitamin A stores in preschool children. Surprisingly, a large percentage of the children were diagnosed with hypervitaminosis A (>1 mol retinol/g liver). This was supported by hypercarotenemia and slightly elevated retinyl ester concentrations. In fact, the children in two of the villages experienced hypercarotenodermia during mango season the following year. These findings were attributed to vitamin A adequate dietary intakes, five years of high-dose vitamin A supplements, and widespread use of vitamin A-fortified sugar. Many African countries are now adopting and mandating vitamin A-fortified foods. In order to ensure that neither deficiency nor hypervitaminosis prevails, countries will need to adopt sensitive RID mass spectrometric methods to assess total body vitamin A stores in carefully selected groups that represent all population strata that are exposed to multiple vitamin A programs. The International Atomic Energy Agency has recently invested in three gas chromatography-mass spectrometers in Africa to further support the use of RID methods on the continent using deuterium and 13C.

Biography:

Pierluigi Mauri is chief of Proteomics and Metabolomics Laboratory ITB-CNR. He has set-up a proteomics laboratory based on the MudPIT (Multidimensional Protein Identification Technology), a gel-free approch (using two dimensional chromatography coupled to tandem mass spectrometry, 2DC-MS/MS) and a parallel computing system for proteomics. This instrumentation allows for the proteomic profiling of biological samples, without limitations, according to to protein molecular weight, pI or hydrophobicity and guarantees high productivity. In addition, the developed computational tools, such as MAPROMA, a label-free quantitation software for simple visualization of data for biologists and clinicians, and extraction of putative biomarkers of diseases or therapeutic treatments. Using this methodology, secreted proteins from tumor (FASEB 2005, Molecular Cancer, 2010) and immune system (JEM, 2011), and from tissues (Im Lett, 2014, Nature Commun 2014) were characterized. Furthermore, direct analysis of biological samples (JMS 2007), enzymatic complexes (Biochimie 2006), development of novel methodological approaches for studying switch-redox proteins (JBC 2005 and 2006, JMB 2006), degenerative diseases (Blood, 2012) and structural and functional characterization of proteins (Biol. Chem. 2004, FEBS 2006) have been performed.

Abstract:

Today, mass spectrometry-based (MS) approach is the leading approach employed in high-throughput analysis. Moreover, the investigation of complex biological samples requires the development of computational tools for processing and handling the great amount of data produced by MS experiments. The recent development of gel-free proteomics approach is allowing a significant improvement over gel-based analysis. Specifically, at ITB-CNR proteomics laboratory, the gel-free MudPIT (Multidimensional Protein Identification Technology) proteomic methodology is used for biomarker discovery and developing clinical methods. Our workflow involves user-friendly tools for characterization of biomarkers and proteotypic peptides, useful for validation step. Specifically, to perform a rapid comparison of protein lists and estimation of differentially expressed proteins, MAProMa (Mauri et al, FASEB J, 2005) home-built software has been developed. It is based on a label-free quantitative approach and it allows the visualization of data in a format more comprehensible to biologists and clinicians(Park et al., Nat methods, 2008; Mauri et Scigelova, Clin Chem Med Lab, 2009). Moreover, the correlation of proteomic data by unsupervised learning algorithms is evaluated; for example, hierarchical cluster groups the protein lists based on information found in the data that describes the objects (proteins) and their relationships (Brambilla et al., JPR 2014). It will be presented technologies and tools to perform clinical proteomics studies involving different biological samples, for investigating human diseases, such as cardio-respiratory, degenerative and tumour diseases.

Biography:

Dr. Pedro Jose Sanches Filho graduated in Degree in Chemistry and graduated in Pharmacy (1988), master's degree in chemistry (1997 ) . Obtained the Ph.D. in Analytical chemistry from the Federal University of Rio Grande do Sul (2002) and post doctoral fellowship by Universidade Nova de Lisboa (2007). He is currently Professor and Researcher at the Instituto Federal de Educação Ciência e Tecnologia Sul-rio-grandense Campus Pelotas -Brazil, leader of the research group on environmental contaminants. Has experience in Analytical Chemistry, with emphasis on Trace Analysis and Environmental Chemistry.

Abstract:

The presence of polycyclic aromatic hydrocarbons (PAHs) and mixture composition in sediments can also be as an indication of sources of pollution. The objective of this study is to evaluate the contamination sources by hydrocarbons derived from urban areas capitated by stormwater drainage systems, and determine the levels of PAHs in the studies channels. The extraction of analytes from sample was made in Soxhlet/ultrasonic bath, and with solvents mixture of hexane and acetone. The fractionation of extracts was performed using a column of Na2SO4, silica and alumina. For the chromatographic analysis, the fractions was injected in the GG-MS in splitless mode. The hydrocarbons were quantified using a GC-MS (Shimadzu QP 2010 ultra), with a RTX 5MS column. Injector and interface were maintained at 2800C and ions source at 2000C, respectively. The electron-impact (EI) ionization energy was 70 eV, and the carrier gas was Helium (99.999%) at a constant flow rate of 1.0 mL min−1 in SIM mode (m/z were, 128, 142, 152, 154, 166, 178, 202, 228, 252, 276, 278). The detection in SIM mode led to a reduction in detection limits and permitted electronic cleanup of the extracts analyzed. We observed that most PAHs detected and quantified showed 4 to 6 rings. The sum of HPAS indicate a moderately impacted environment, with a predominance of pyrolytic sources. This calls attention to the impact of runoff from urban areas under storm drainage channels, leading to these pollutants area of great environmental importance.

Biography:

Pawel Jaruga received his Ph.D. in Medical Biology from Medical University in Bydgoszcz, Poland, in 1994 and D.Sc. degree from Collegium Medicum of Nicolaus Copernicus University in Torun, Poland in 2004. He held postdoctoral fellowships at Chemical Science Laboratory at National Institute of Standards and Technology (NIST) (1994-1996, 2000-2002). Between 2002-2010, he remained affiliated with NIST as Senior Research Scientist at Department of Chemical and Biochemical Engineering, University of Maryland, Baltimore, MD and then as Senior Life Scientist at Science Applications International Corporation, Homeland Protection and Preparedness Business Unit, Washington, DC. In 2010, he was appointed as a Research Chemist at NIST in the Biomolecular Measurement Division. He published 87 papers in peer-reviewed journals.

Abstract:

Among cellular structures, the genome is particularly prone to damage, which can result from spontaneous reactions, replication linked failures, or oxidative processes due to metabolic derivatives or to external agents. Damage to DNA causes more severe consequences than damage to replaceable cellular macromolecules because the genome must be preserved for the life of the cell, and because it can be copied and proliferated into next generations of the cells. Reactions of free radicals and other redox capable agents with DNA generate an abundance of products in nuclear and mitochondrial DNA of living organisms. Growing evidence points to the involvement of this type of damage in the etiology of numerous diseases including carcinogenesis. Comprehensive understanding of the mechanisms, cellular repair, and biological consequences of DNA damage requires accurate measurement of resulting products. There are various analytical techniques, with their own advantages and drawbacks, which can be used for this purpose. Mass spectrometric techniques with isotope dilution provide structural interpretation of products and accurate quantification, which ascertain reliable measurement. Gas chromatography-mass spectrometry or liquid chromatography-mass spectrometry, in single or tandem versions, have been used for the measurement of numerous DNA products such as sugar and base lesions, 8,5′-cyclopurine-2′-deoxynucleosides, base-base tandem lesions, and DNA-protein crosslinks, in vitro and in vivo. Basic concepts and results will be presented and discussed.

Biography:

Mehran F. Moghaddam is associate professor in Celgene Corporation, USA. He has done work to manage compound attrition and to improve the odds of discovering viable development candidates, medicinal chemistry departments have increased their productivity and produce an ever increasing number of compounds. Most compounds are evaluated in a battery of in vitro and in vivo assays in drug metabolism and pharmacokinetics departments. He has published more than 50 research paper.

Abstract:

In modern drug discovery, mass spectrometry (MS) plays a central role in determination of drug-like properties of small molecules. In an effort to manage compound attrition and to improve the odds of discovering viable development candidates, medicinal chemistry departments have increased their productivity and produce an ever increasing number of compounds. Most compounds are evaluated in a battery of in vitro and in vivo assays in drug metabolism and pharmacokinetics (DMPK) departments. These are labor intensive and time consuming assays and almost always utilize qualitative or quantitative mass spectrometric methods. DMPK groups have to mirror the increased productivity exhibited by the medicinal chemistry to prevent bottlenecks. In addition to assembling a highly qualified team and automation, a DMPK department must be creative in the use of its resources and the utility of MS in order to enabled generation of high quality data in an efficient, timely, and cost effective manner. Qualitative and quantitative MS approaches and their specific utility in discovery DMPK will be discussed in this presentation.

Biography:

Jun cao is associate professor in Hangzhou Normal University, China. He has done work for a rapid, simple, optimized, and validated miniaturized SPE method for the separation and quantification of six flavanones in Citrus fruit extracts by ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry using a dual electrospray ionization source. He has published more than 50 papers.

Abstract:

Traditional solid phase extraction (SPE) method requires large volumes of elution solvent and high consumption of sorbents, making the whole procedure expensive and time consuming. Therefore, to establish a reliable miniaturized SPE approach for the extraction of tested compounds from complex samples is quite meaningful. Moreover, mesoporous molecular sieves have not been explored extensively as a sorbent for miniaturized SPE. In particular, its applications for the extraction of trace compounds from complex fruits are scarce, barring work from our group. This article reported a rapid, simple, optimized, and validated miniaturized SPE method for the separation and quantification of six flavanones in Citrus fruit extracts by ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry using a dual electrospray ionization source. The mesoporous molecular sieve SBA-15 as a novel solid sorbent for the extraction of target analytes was characterized by Fourier transform-infrared spectroscopy and scanning electron microscopy. Additionally, compared with other reported extraction techniques, the mesoporous SBA-15 based SPE method possessed the advantages of shorter analysis time and higher sensitivity. Furthermore, considering the different nature of the tested compounds, all of the parameters, including the SBA-15 amount, solution pH, elution solvent, and the sorbent type, were investigated in detail. Under the optimum condition, the instrumental detection and quantitation limits calculated were less than 4.26 and 14.29 ng mL-1, respectively. The recoveries obtained for all the analytes were ranging from 89.22% to 103.46%. The experimental results suggest that SBA-15 is a promising material for the purification and enrichment of target flavanones from complex citrus fruit samples.

Biography:

Joydip Das is assistant professor in University of Houston, USA. He has investigated alcohol binding sites in protein kinase C (PKC) epsilon, a serine/threonine kinase and Munc13-1, a presynaptic protein expressed predominantly in the brain. To identify alcohol binding site. He has published more than 60 research papers.

Abstract:

Alcohols bind and regulate functions of many proteins in the human brain. To develop antagonists for the alcohol addiction and alcoholism, it is necessary to identify the site of action of alcohol in these proteins. We have investigated alcohol binding sites in protein kinase C (PKC) epsilon, a serine/threonine kinase and Munc13-1, a presynaptic protein expressed predominantly in the brain. To identify alcohol binding site(s), C1 domains of PKC and Munc13-1 were photolabeled with the diazirine analogs of butanol and octanol, 3-azibutanol and 3-azioctanol, and the sites of photoincorporation were identified by MS/MS analysis using electrospray ionization mode. Azialcohols labeled Tyr-176 of PKCC1A, His-248 and Tyr-250 of PKCC1B. In Munc13-1, Glu-582 is exclusively photolabeled by the azialcohols. Azialcohols failed to photolabel these sites when these residues were mutated with alanine. Inspection of the model structure of PKCC1 and Munc13-1C1 reveals that these residues forms groove where alcohol can bind. The present results provide evidence for the presence of alcohol-binding sites on PKC and Munc13-1 and underscore the power of mass spectrometry in identifying sites of alcohol action in the brain.

Biography:

Eduarda Silva carried out her PhD studies, concerning the total synthesis of mycaperoxide B probing a biomimetic approach, under the guidance of Professor Laurence Harwood at University of Reading (UK). She finished her PhD degree in 2009 and joined the group of Professor Artur Silva as postdoctoral fellow at University of Aveiro. Her work is now concerned with catalysis, stereochemistry and the formation of carbon-carbon bonds by 1,6-conjugated addition. She is also interested in the identification and detailed structural information of new synthesized and bioactive compounds attained by mass spectrometry. She has published 20 papers in international peer review journals.

Abstract:

Xanthones (XH) are a class of heterocyclic compounds widely distributed in nature holding numerous noteworthy biological and antioxidant activities. Therefore, it is important to achieve the utmost detailed structural information relevant to understand and predict their biological properties. In these studies we aimed to explore the potential relationship between radical-mediated xanthone chemistry in the gas phase and their antioxidant activities. The mass spectral fragmentation behavior of nine 2,3-diarylxanthones bearing up to four hydroxyl groups at various sites on the two aryl rings was surveyed using electrospray ionization tandem mass spectrometry. Protonated XH were generated in the gas phase by electrospray ionization (ESI) and the main fragmentation pathways of the protonated XH formed due to collision-induced dissociation (CID) were investigated. In the CID-MS/MS spectra of [M+H]+ ions of six of the studied xanthones, showing a most promising biological profile, the product ion produced with the highest relative abundance (RA) corresponded to the one formed through concomitant loss of H2O plus CO. The product ion formed by loss of 64 Da (concomitant loss of two molecules of H2O plus CO) is only observed for xanthones containing a catechol unit. This product ion has the highest RA for the most potent scavenger of reactive oxygen and nitrogen species containing two catechol moieties. Through this studies a strong relationship between some of the biological activities of the studied 2,3-diarylxhantones and their ESI-MS/MS fragmentation spectra was found. The multivariate statistical analysis results suggest that the selected MS features are related to the important biological features.

Biography:

Dr. Dizdaroglu has obtained his PhD at the Karlsruhe Technical University, Germany, and subsequently worked for seven years at the Max-Planck-Institute for Radiation Chemistry, Germany, before moving to US in 1978. He has been at the National Institute of Standards and Technology (NIST) for more 30 years. In 2006, Dr. Dizdaroglu was conferred upon the rank of NIST Fellow. He published more than highly cited 230 papers. Dr. Dizdaroglu received numerous scientific awards including the Hillebrand Prize of the American Chemical Society and the Gold Medal Award of the US Department of Commerce. He was also awarded two Honorary Doctorates.

Abstract:

Malignant tumors possess increased DNA repair capacity that may affect the therapy and outcome of cancer. Thus, DNA repair proteins are becoming predictive, prognostic and therapeutic factors in cancer. Accurate measurement of expression levels of these proteins in tumors and normal tissues will likely help develop and guide treatment strategies. We developed an approach involving LC-MS/MS with isotope-dilution to positively identify and accurately quantify several DNA repair proteins in human cells, including APE1 and MTH1. As a major endonuclease in mammals, APE1 is involved in base excision repair and possesses other functions. Evidence points to the predictive and prognostic value of APE1 expression and subcellular localization in human cancers. MTH1 sanitizes the nucleotide pool so that oxidatively modified 2'-deoxynucleoside triphosphates (dNTPs) cannot be used in DNA replication. Cancer cells require MTH1 to avoid incorporation of modified dNTPs resulting in DNA damage leading to apoptosis. We produced 15N-labeled full-length human APE1 and MTH1 be used as internal standards. Unlabeled and 15N-labeled proteins were digested with trypsin and analyzed by LC-MS/MS. Numerous tryptic peptides of APE1 and MTH1 were identified by their mass spectra. Multiple-reaction monitoring was used to monitor characteristic mass transitions of peptides. Subsequently, APE1 and MTH1 were identified and quantified in cultured human cell lines, and in human normal and cancerous breast tissues. Overexpression and subcellular de-localization in cancer cell lines and breast cancer tissues were observed. The novel approach developed in this work may help elucidate the role of APE1 and MTH1 in disease development and treatment responses.

Biography:

Shawn R. Campagna received his Ph.D. from Princeton in 2006, after working with Prof. Martin Semmelhack on a joint project with Profs. Bonnie Bassler and Frederick Hughson to characterize the chemical properties of an interspecies bacterial signaling molecule, autoinducer-2. He then performed post-doctoral research with Prof. Joshua Rabinowitz at the Lewis-Sigler Institute for Integrative Genomics at Princeton University where he developed mass spectrometric methods for the identification of novel biochemical pathways and natural products from whole cell extracts. He joined the Chemistry Department at UT Knoxville in 2007 where he has worked to develop new metabolomic and lipidomic techniques. Dr. Campagna is currently an Associate Professor and Head of the UTK Biological and Small Molecule Mass Spectrometry Core.

Abstract:

Recent advances in liquid chromatography–mass spectrometry (LC–MS) based metabolomics and lipidomics have furthered understanding of medicinally and environmentally relevant microbial processes. The utility of metabolomics methods is enhanced by monitoring the incorporation of stable isotope-labeled nutrients, either 15N or 13C, into the metabolome using kinetic flux profiling techniques (KFP). The coupling of metabolite concentration (pool size) measurements and KFP can be used to determine both the amounts of metabolites within and relative rates of flux through many biochemical pathways in vivo, and the combination of these methods allow a global snapshot of cellular metabolism to be obtained. Our laboratory’s primary analytical platform is a set of UPLC--Orbitrap MSs with electrospray ionization sources, and the methods employed attempt to measure pool size of a large set of analytically tractable water and lipid soluble metabolites from all kingdoms of life. The set of methods measure at least one metabolite from all known carbon and nitrogen utilization pathways, the activated methyl cycle, all amino acid and nucleotide biosynthesis pathways, as well as lipids with diverse head groups. Several vignettes from our work studying aquatic microorganisms will be discussed, with a focus on understanding the utility of metabolomics to understand mixed populations of viruses, bacteria, cyanobacteria, and algae. The information gained from these meta-metabolomics experiments is often enhanced when coupled with other systems biology tools, such as meta-transciptomics; and the integration of multi-omics techniques to study the metabolism of aquatic microbial consortia will be discussed.

Biography:

Fernando Ramos is Associate Professor of the Pharmacy Faculty of Coimbra University and Senior Research of the CNC – Center for Neuroscience and Cell Biology. He has published more than 75 scientific publications, among books, book chapters and national and international papers, most of them in the field of drug residue analyze using Mass Spectrometry

Abstract:

Food safety is one of the top priorities assumed worldwide by several international organizations. The European Commission has been continuously revising and implementing legislation in order to control food from “farm to fork”. One of the constant concerns is related with inappropriate and abusive use of veterinary drugs, in food-producing animals, as growth promoters. A large number of different families of compounds can be included in such treatments being its principal route of administration by feed and drinking water. To protect the consumers from undesirable health problems, the development of analytical methods for the determination of veterinary drug residues in food of animal origin, destined to human consumption became a mandatory issue in Food Safety. In terms of analytical detection, mass spectrometry is considered a highly selective technique that provides the indispensable feature of accurate identification of the substance present in the biological samples. Mass spectrometry, as triple quadrupole coupled with liquid chromatography (LC-MS/MS) is a powerful tool that allows not just single determinations but also multi-compound detection by recording full mass spectra (scan mode), selected ion monitoring (SIM) and multiple reaction monitoring (MRM). More recently, it started to grow the application of Time-of-Flight (ToF) or High Resolution Mass Spectrometry (HR-MS) in residues analysis. Nevertheless, the potential of mass spectrometry metabolomics for veterinary drug residue analysis and chemical food safety in general will be also discussed.

Biography:

Anthony N. Gachanja is associate professor in Jomo Kenyatta University of Agriculture and Technology, Kenya.He has work on current level of GC-MS instrumentation in public universities, government and private laboratories, with highlights on state-of-art equipment, will be discussed. Challenges in acquisition, installation and routine usage of gas chromatography-mass spectrometry (GC-MS) instrumentation in Eastern Africa region. He has published more than 50 research papers.

Abstract:

The availability and usage of analytical instrumentation is steadily improving in research, private and institutional laboratories within the Eastern African region. It is critical for governments and institutions to make decisions on developmental areas (e.g. millennium development goals on health, food security, water and industrialization etc) using analytical data from validated methodologies using best available instrumentation. The current level of GC-MS instrumentation in public universities, government and private laboratories, with highlights on state-of-art equipment, will be discussed. Challenges in acquisition, installation and routine usage of gas chromatography-mass spectrometry (GC-MS) instrumentation in Eastern Africa region will be presented. With the support of Pan-African chemistry Network and The Royal Society of Chemistry, training workshops have been held in the region for GC-MS users. Lessons learnt through these workshops will be discussed. The experiences and current application of GC-MS with examples such analysis of organic pollutants in water, profiling of honey from different origins, and organic contaminants in vegetables available in Kenyan market. Suggestions on the way forward for advancement of GC-MS and MS in the region will be presented.

Biography:

Tania Petta has completed her Master in Chemistry from University of São Paulo (USP, Brazil) in 2008. She completed her PhD in Chemistry from USP in 2012, at the age of 29 years. At present, she is a post-doc fellow at USP, at the Department of Pharmaceutical Sciences. She has experience in LC-MS/MS, microbial secondary metabolites isolation, biotransformation, and lipid analysis. Her current research focuses on mass spectrometry-based lipidomics applied to study the role of lipid mediators during infections.

Abstract:

Polyunsaturated fatty acids (PUFA) such as arachidonic acid (AA-C20:4), eicosapentaenoic acid (EPA-C20:5) and docosahexahenoic acid (DHA-C22:6) are precursors in the biosynthesis of bioactive oxylipids known as eicosanoids and docosanoids, which are involved in many immunological and physiological processes (1). The position at which an oxygen is inserted into the PUFA chain is highly controlled by regio- and stereoselective enzymes, such as LOX, COX and cytocrome-P450 (2). In biological systems, they are minor components present as isomeric and isobaric species. In this sense, Mass Spectrometry (MS) have shown great potential for quantification and identification of these lipid species (3). In this work, we present the versatility of tandem mass spectrometry for target and untargeted analysis of oxylipids. The developed methods were applied to analyze a hypothalamus rat extract. Using the target LC-MS/MS approach, 10 pre-selected eicosanoids were identified in the crude extract. The untargeted LC-MS/MS approach enabled the detection of several mono- and poly-oxygenated AA products, which were not identified when using the targeted method. Their structures were proposed according to individual product-ion mass spectra achieved via collision induced dissociation. In conjunction, these MS methods represent powerful strategies to accelerate the identification of oxylipids, and can strongly contribute to the discovery of novel lipid mediators in biological samples.

Biography:

Vladimir Zaichick is a Nuclear Physicist, Biologist and Researcher. He obtained his MS in 1966 from the Moscow Institute of Engineering Physics, his PhD (nuclear physics) in 1972 from the Institute of Biophysics, Moscow, and his DSc (radiobiology) in 2011 from Medical Radiological Research Center, Obninsk, Russia. He is a full professor of radiobiology, a fellow of the British Royal Society of Chemistry (FRSC) and a Chartered Chemist (CChem) since 1996, a Presidium Member of the Scientific Council on the Application of Nuclear Methods and a member of the Scientific Council on Analytical Chemistry of the Russian Academy of Sciences. He has made 137 presentations at seminars, published more 300 papers in reputed journals and has been serving as an editorial board member.

Abstract:

Since the times of the alchemists, chemical elements have been investigated in human organs, tissues and fluids. During the last decades the number of publications devoted to them increased considerably. Today, the number of published articles may be estimated at about twenty thousands, and the amount of monographs about hundred. This vast amount of data, dealing with the importance of elements acquired in different fields of scientific research and practical life, puts forward the need for a synthetic approach in element research. At the end of the 20th century, a new scientific discipline appeared, focused on the role of chemical elements in human body under physiological and pathological conditions. This new field of interdisciplinary study has been named: “Medical Elementology” (from lat. “medicina” and “elementum”). A lot of medical doctors, toxicologist, ecologist, chemists and physicist who were involved in the study of chemical elements in medicine and biology had thus the feeling that they were working in new self-sufficient scientific sphere. As a rule, all scientific disciplines are characterized, first of all: 1) by the subject of study; 2) by accepted postulates; 3) by research methods; 4) by methods of quality control; 5) by terms and definitions. Instrumental analytical methods such as non-destructive NAA and EDXRF as well as destructive AAS, ICP-AES and ICP-MS are the main research instruments in Medical Elementology. Role and place of ICP-MS will be discussed using our results obtained in the age-dependence studies of 67 chemical element contents in human bone, hair and prostate gland.

Biography:

Xiao Ding has completed his PhD in Analytical Chemistry in 1997 and postdoctoral studies in 1999 from Department of Chemistry in University of Arizona. She has been working in the bioanalytical field to support drug discovery and development for more than 15 years. Currently she is a senior scientist in the department of metabolism and pharmacokinetics in Genentech (Roche), a pharmaceutical company. She has published more than 25 papers in reputed journals and presented more than 40 posters at major conferences.

Abstract:

GDC-0032 (taselisib) is a potent, selective small-molecule inhibitor of Class I phosphoinositide 3-kinase (PI3K). It is being developed by Genentech for the treatment of various malignancies and is currently in multiple clinical trials. An accelerator mass spectrometry (AMS) method for the determination of 14C-GDC-0032 concentrations in human plasma was developed and qualified for the first time according to the Guidance for Industry: Bioanalytical Method Validation issued by the Food and Drug Administration (FDA). The absolute bioavailability of GDC-0032 was determined following oral administration of 3-mg GDC-0032 and IV administration of 3-μg/200 nCi 14C-GDC-0032 in male healthy volunteers in a clinical trial. UPLC-AMS method provided high selectivity and high sensitivity due to its nature of measuring the absolute 14C label. The method was qualified over the calibration curve range 0.168 to 100.8 pg/mL (0.025 to 15.0 dpm/mL) using linear regression and 1/y² weighting. Within-run relative standard deviation (%RSD) ranged from 1.87 to 8.59%, while the between-run %RSD varied from 3.21 to 7.29% for QCs using a standard curve model. The accuracy ranged from 98.60% to 109.93% of nominal for within-run and 100.96% to 106.95% of nominal for between-run at all concentrations. Extraction recovery factor of 14C-GDC-0032 was 0.9050. Stability of 14C-GDC-0032 was established in human plasma for 51 days at -70 ï‚°C and established in reconstituted sample extracts for 20 hours at -70 ï‚°C. The absolute bioavailability of GDC-0032 capsules was 57.4% (34.6% - 80.3%).

Biography:

Scott M. Grayson is associate professor in Tulane University, USA.He has done work on Traditional characterization techniques such as mass spectrometry can provide absolute molecular weight, while size exclusion chromatography can enable a determination of the hydrodynamic radius. By comparing these data, along with additional techniques such as viscometry.He has published more than 60 research papers.

Abstract:

The ability to characterize and differentiate polymers of diverse architecture or topology remains a challenge among polymer scientist. Traditional characterization techniques such as mass spectrometry can provide absolute molecular weight, while size exclusion chromatography can enable a determination of the hydrodynamic radius. By comparing these data, along with additional techniques bsuch as viscometry, the extent to which a polymer exhibits a more collapsed or extended confirmation can be determined. However, ion mobility spectrometry-mass spectrometry (IMS-MS) provides an attractive alternative to these approaches, as this single technique simultaneously separates the components of a polymer sample by both drift time (which correlates to the size for a given charge state) and molecular weight. The resolution provided in both of these dimensions is beneficial not only for determining the macromolecular architecture, but also in the more challenging problem of determining architectural purity. The application of IMS-MS to elucidating polymer architecture will be probed with a number of case studies including cyclic, branched, and linear polymers.

Biography:

Steven J. Bark is associate professor in University of Houston, USA. He has done work for successful use of tagging approach is well represented in current literature. However, we have observed that the matrix used to immobilize the antibody is critical for successful affinity purification. Initial observations suggested high variability between experiments using the identical M2 Clone antibody and p53 FLAG-tagged protein. He has published more than 30 research papers.

Abstract:

Affinity Purification Mass Spectrometry (AP-MS) is a highly effective method for isolating and identifying binding partners to a target protein. One of the most common methods used for AP-MS experiments is expressing the target protein with a unique peptide sequence tag such as FLAG, c-Myc, HA or V5 and using the well documented high affinity antibodies to these peptide sequences. The successful use of this tagging approach is well represented in current literature. However, we have observed that the matrix used to immobilize the antibody is critical for successful affinity purification. Initial observations suggested high variability between experiments using the identical M2 Clone antibody and p53 FLAG-tagged protein. We have experimentally evaluated several different resin formats based on these reagents and identified multiple parameters underlying the observed variability in the affinity purification procedure. Additionally, crosslinking procedures, reagents, and dilution conditions with different detergent additives were found to differentially affect these resins and have critical impact on the success of the AP-MS experiment. Examples of successful AP-MS experiments after optimization of these parameters will be presented.

Biography:

Sihe Wang, PhD is Section Head and Medical Director of Clinical Biochemistry and Director of Clinical Biochemistry Fellowship Training Program at Cleveland Clinic. He chairs clinical chemistry integration team for the Cleveland Clinic Health System which includes one Florida hospital, eight community hospitals and eighteen family health centers in Northeast Ohio. Additionally, he is a Clinical Chemistry Professor, Cleveland State University. His expertise includes general clinical chemistry and clinical application of mass spectrometry. Sihe is a diplomate of the American Board of Clinical Chemistry since 2005 and a fellow of the National Academy of Clinical Biochemistry since 2006. Sihe

Abstract:

Based on the 2011 report by Institute of Medicine, pain is a national issue with significant social and economic consequences. Therefore, relieving pain should be a national priority. Recently pain management has drawn much attention due to the high potential of pain medication diversion and drug abuse. As such laboratory testing plays a critical role in assisting physicians to monitor patient compliance. Immunoassays are quick but not the ideal tools due to the false negative and false positive results. Liquid Chromatography-tandem mass spectrometry (LC-MS/MS) has been increasingly used for pain management testing due to its high specificity and sensitivity. In this presentation challenges associated with pain management and laboratory testing to support pain management will be reviewed. An example of a comprehensive method for analyzing multiple drugs/metabolites in urine by LC-MS/MS will be discussed in detail.

Biography:

Frédéric Aubriet has completed his PhD at the age of 27 years from University of Lorraine (France). He obtained an associated professor position in 2001 and a full professor position in 2012 at the University of Lorraine. He manages a research team focusing on the use of mass spectrometry and especially FTICRMS in the field of complex system analysis. His main research interests are dedicated to the analysis of indoor, outdoor and industrial pollution and second generation biofuels. He has also a significant expertise in the field of inorganic mass spectrometry and ion-molecule reaction.

Abstract:

Nowadays, the analysis of complex systems for different purposes is a great challenge. Two methodologies based on mass spectrometry may be used. The first one involves a targeted approach, which requires pretreatment steps including extraction, purification and separation (liquid or gas chromatography) before mass spectrometry measurement. This methodology is well suited for the quantization of targeted compounds but does not allow a global description of the sample to be achieved. Alternatively, the non-targeted methodology introduced few years ago may be used. Both approaches ensure to obtain complementary information. Non-targeted approach allows an exhaustive description of the complexity of the sample to be obtained, but it does not allow quantization. In contrast, non-targeted methodology ensures quantization but only on a restricted number of compounds. In contrast to the mass spectrometry targeted methods, the non-targeted approach systematically requires ultra-high resolution mass spectrometry, which allows the global composition of complex mixtures to be accurately detailed. Moreover, the ability of this instrument to be coupled with various ionization sources increases its capability to detect compounds with very different chemical properties and consequently to obtain an accurate description of the investigated sample. Considering the important analytical capacities of this approach, we will present two applications of this method. The first one in the field of environment chemistry for the analysis of particulate matter from different cigarette smokes which are some of the more important indoor pollutants and the second one in the field of the characterization of bio oils produced by pyrolysis of the biomass.

Biography:

Fedyunina Natalia has defended her PhD in 2013 at the age of 25 years from Lomonosov Moscow State University. She has published 5 papers in reputed journals. Now she works at National University of Science and Technology "MISIS" through the “Program to Increase the Competitiveness of NUST MISIS Among World-Leading Scientific Educational Centers”. She is senior researcher in Laboratory of separation and preconcentration in chemical diagnostics of functional materials and environmental objects, project of which has been recognized as promising at International Scientific Committee with the participation of leading Cambridge, Stanford, Technion scientists.

Abstract:

Inductively coupled plasma mass spectrometry (ICP-MS) is the most widely used technique for determination of rare earth elements (REEs) and nobel metals (NMs) in different types of rock samples. The rock sample digestion stage should provide the completeness of the analyte transfer from a solid sample to liquid phase and, if possible, the minimization of the matrix elements concentration in the final solution. The concentration of elements in the solution to be analyzed after digestion of rock samples is reduced by one to two orders of magnitude. Besides, the spectral and non-spectral interferences caused by high concentrations of matrix elements reduce the sensitivity of mass spectrometric determination. Therefore, the preconcentration stage is usually needed. Different methods of rock sample digestion for final analysis by ICP-MS technique were investigated. To avoid problems with the introduction in ICP (the variation of plasma temperature, contamination of the sample introduction system and clogging of the cones) of high total dissolved solid concentrated solutions after fusion and compensate the loss of sensitivity preconcentration using Pol-DETATA (diethyltriaminetetraacetate), hyper-crosslinked polysterene and StrataX-AW (with ethylenediamine groups) sorbents were tested. Flow-injection mode was used for organic and high acidity eluates introduction to ICP-MS.

Biography:

Prof. Hans-Arno Synal graduated in Physics at the University of Bonn (Germany). In 1986, he moved to ETH Zurich (Switzerland) for his PhD studies. As research scientist, at the Paul Scherrer Institute (Switzerland), he became group leader and responsible for the accelerator facilities at the Ion Beam Physics Laboratory. In 2008, he was appointed as head of the ETH Laboratory of Ion Beam Physics and since 2011 he is honorary Professor at ETH Zurich. He has published more than 230 papers in reputed journals.

Abstract:

The technical evolution of Accelerator Mass Spectrometry (AMS) instrumentation over the last ten years is summarized. A key characteristic of AMS is destruction of molecular interferences and subsequent analyses of atomic ions. It makes the extreme abundance sensitivity possible (in specific cases, below isotopic ratios of 10-16). This can be reached with instruments having quite modes mass resolving power (M/∆M less than 300). Today, 1+ charge state is used, molecular interferences are destroyed in multiple collisions with stripper gas atoms or molecules, and a high yield atomic ions is reached at energies of a few hundred keV. Thus, AMS instruments develop towards lab size or tabletop devices. The use of He as stripper gas has further improved performance with respect to overall detection efficiency and reproducibility of measurement conditions. In parallel, implementation of permanent magnets into dedicated radiocarbon AMS system is progressed. This reduces complexity of the instruments and significantly reduces operation and installation costs. For radiocarbon, He stripping has potential to further down size instruments and reduce the ion energy below 50 keV. I will summarize the latest achievements. But, low energy AMS is not limited to radiocarbon only and there is a great potential for 10Be, 26Al, 129I and actinides measurements at compact AMS systems. These developments have launched the wide spread use of AMS in various research fields and has resulted in a boom of new AMS facilities. The related impact to the wide variety of applications of AMS in modern research is not covered.

Biography:

Abstract:

Lipids are a large and diverse group of naturally-occurring compounds that store energy, give structure to cell membranes, and, as ´molecular messengers´, they serve as signaling molecules. Considering foods nutritional value, lipids are one of the key components, they are e.g. a source of essential components such as polyunsaturated fatty acids (PUFA), they support absorption of fat soluble vitamins etc. For in-depth structural and quantitative characterization of various lipid classes and their distribution within food crops and/or products, instrumental platforms based on advanced chromatographic methods coupled with mass spectrometric detection represent currently the most challenging option. In our studies, we focused on application of these novel strategies for monitoring of lipids changes during storage and processing, i.e. under conditions when they may undergo various degradative processes such as oxidation. Worth to notice that compounds generated through oxidation reactions are related to undesirable sensory and biological effects. In addition to cytotoxic and genotoxic compounds, free radicals, products of lipids peroxidation also co-oxidize some vitamins, and thereby impair the nutritional quality of the foods. A high number of methodologies enabling determination of both primary and secondary oxidation products has been developed and implemented. However, some of these ´classic´ approaches such as peroxide value illustrating early stages of lipids oxidation, are highly empirical and their accuracy is rather questionable since the results vary with details of the analytical procedure. Also thiobarbituric acid (TBA) test often applied for measurement of the extent of lipids oxidation has been criticized as being non-specific and insensitive for the detection of low levels of malonaldehyde. Not surprising that introduction of modern instrumental techniques that enable rapid obtaining of more comprehensive and specific information is urgently needed. In addition to implementation of modern analytical methods for assessment of lipids qualitative parameters, we also were concerned with introduction of novel non-target screening strategies enabling lipids authentication. This presentation involves several case studies documenting challenges in lipids analysis:• Simple and fast sample preparation strategy based on partition in ternary solvent system enabling, in a single step, fractionation of fish lipid classes according their polarity. • Application of an ambient high resolution mass spectrometry (HRMS) employing direct Analysis in Real Time (DART) ion source for an assessment of lipids quality with regards to content of primary and secondary oxidation products • Authentication of food lipids using fingerprinting / profiling strategy based on DART-HRMS, lipidomics • Employing supercritical fluid chromatography (SFC) coupled with high definition mass spectrometry (HDMS) for analysis of frying oils and lipids isolated from human adipose tissue. Challenges offered by metabolomics in nutrition research are thoroughly discussed with a special focus on application of advanced multivariate statistical strategies enabling data interpretation.

Biography:

Abstract:

Emission of carbon from ecosystems in the form of volatile organic compounds (VOC) represents a minor component flux in the global carbon cycle that has a large impact on ground-level ozone, particle and aerosol formation and thus on air chemistry and quality. Moreover, combustion of solid and liquid fuels is the largest source of potentially toxic VOC, which can strongly affect health and the physical and chemical properties of the atmosphere. Among combustion processes, biomass burning is one of the largest at global scale. We used a Proton Transfer Reaction “Time-of-Flight” Mass Spectrometer (PTR-TOF-MS), which couples high sensitivity with higher mass resolution, both in combination with the eddy covariance method to measure by-directional fluxes of VOC in a poplar-based short rotation coppice (SRC) plantation, and for real-time detection of a multiple VOC emitted by burned hay and straw in a barn located near our measuring station. Whereas abundant fluxes of isoprene, methanol and, to a lesser extent, fluxes of other oxygenated VOC such as formaldehyde, isoprene oxidation products (methyl vinyl ketone and methacrolein), methyl ethyl ketone, acetaldehyde, acetone and acetic acid, were exchanged between the poplar plantation and the atmosphere, we detected 132 different organic ions directly attributable to VOC emitted from the fire. Methanol, acetaldehyde, acetone, methyl vinyl ether (MVE), acetic acid and glycolaldehyde dominated the VOC mixture composition. The time-course of the 25 most abundant VOC, representing ~ 85% of the whole mixture of VOC, was associated with that of carbon monoxide (CO), carbon dioxide (CO2) and methane (CH4) emissions. The strong linear relationship between the concentrations of pyrogenic VOC and of a reference species (i.e. CO) allowed us to compile a list of emission ratios (ERs) and emission factors (EFs), but values of ER (and EF) were overestimated due to the limited mixing of the gases under the stable (non-turbulent) nocturnal conditions. In addition to the 25 most abundant VOCs, chemical formula and concentrations of the residual, less abundant VOCs in the emitted mixture were also estimated by PTR-TOF-MS. Furthermore, a time-resolved evolution of the complex combustion process was described on the basis of the diverse types of pyrogenic VOCs recorded.

Biography:

Bernard Do has completed his PhD at the age of 29 years from Paris-Descartes University (France). He is hospital pharmacist, associate professor and senior researcher of a research team focusing on drug intrinsic stability and drug/polymer interactions, at Assistance Publique-Hôpitaux de Paris and Paris-Saclay University. He has published more than 40 papers in reputed journals and serving as an editorial board member of repute.

Abstract:

As the active pharmaceutical ingredient (API) may undergo degradation, leading to the drug activity loss or to occurrence of adverse effects associated with degradation products, thorough knowledge of API’s stability profile is required. Liquid chromatography coupled to multi stage high-resolution mass spectrometry has shown suitable for the characterization of drugs degradation pathways. Such an approach has been successfully applied to the study of the behaviour of a new anticoagulant in the presence of various stress conditions. Dabigatran etexilate (DABET) is an oral direct thrombin inhibitor that has been approved for the prevention of blood clot formation. Since very few study has been reported on the drug stability profile, a study related to DABET’s behaviour under stress conditions was carried out in order to identify its major degradation pathways. DABET was subjected to hydrolytic (acidic and alkaline), oxidative, photolytic and thermal stress, as per ICH-specified conditions. The degradation products formed were separated by liquid chromatography (LC) using Kinetex™ 2.6 µm C18 100 A°, 50 x 2.1 mm LC column and multistage gradient mobile phase composed of water and methanol. Up to ten degradation products along with dabigatran, the active metabolite of DABET, were detected and characterized by studying their fragmentation patterns in high-resolution mass spectrometry (ex: Figure 1). Under hydrolytic stress conditions, O-dealkylation may occur and formation of benzimidic acid derivatives was also observed. DABET was shown much less susceptible to photolysis and oxidative stress, even if N-dealkylation was highlighted.

Biography:

Claire Lemaire is biochemist specialized in membrane proteins. She began her career in the photosynthesis field on the assembly and regulation of photosynthetic complexes (I.B.P.C. ,Paris). She then joined the C.N.R.S. (French National Center for Scientific Research) where she acquired a solid expertise in the respiratory complexes. Over the last six years, she has developed a research project with her group focusing on the regulation of OXPHOS complexes by phosphorylation.

Abstract:

The mitochondrion is an organelle of which the most important function is to provide energy to the cell generated by oxidative phosphorylation catalyzed by the respiratory enzymes. In humans, deregulation of mitochondrial functions is associated with several pathologies. The activity of the respiratory enzymes may be modulated in response to metabolic demand and various types of stress. Several levels of regulation may be conceived, including post-translational modifications, such as phosphorylation. The steadily increasing number of identified mitochondrial phosphoproteins suggests that reversible protein phosphorylation could be an important level of regulation in mitochondria. However, this hypothesis cannot be tested without quantitative data on variations in the abundance of mitochondrial proteins and their level of phosphorylation under different growth conditions. The yeast Saccharomyces cerevisiae is a powerful tool for studying various energetic and physiological states. We present for the first time a quantitative study of both protein abundance and phosphorylation levels in isolated yeast mitochondria under respiratory and fermentative conditions. To focus our analysis specifically on mitochondrial proteins, we performed a subcellular fractionation and used LC–MS/MS to overcome the limitations of 2D gel electrophoresis. Protein abundances were quantified using a label-free method. The phosphoproteome was analyzed quantitatively using the multiplex stable isotope dimethyl labeling procedure. For all quantified phosphopeptides, protein abundance was determined, allowing normalization of the data and permitting analysis of the specific variation of phosphorylation status independent of changes in protein abundance. This study provided reliable information on how the yeast mitochondrial proteome and phosphoproteome adapt to different carbon sources.

Biography:

Isabelle Silvis started her PhD in 2013 RIKILT in the authenticity group of Saskia van Ruth, in which she is conducting research on food fraud of herbs and spices. The focus is on the development of analytical methods for the detection of economically motivated adulterations. In 2013 she obtained her European Master’s degree in Food Technology, from Wageningen University. Her international background is quite useful in her current activities as her project is part of SPICED, in which European partners from different universities, institutes and industry build on a strong consortium, to secure the spice and herb chain from primary production to consumer-ready food against major deliberate, accidental or natural CBRN contamination.

Abstract:

Abstract Due to several recent food fraud scandals there is an increased interest of industry and consumers to obtain relevant and reliable information about the integrity of food products. The food commodity ‘herbs and spices’ are situated in the top 10 of most adulterated products world-wide. Therefore in this study, we explored direct mass spectrometry techniques in analysing authentic spices to create base-line fingerprints of genuine herb and spice material. Untargeted fingerprinting techniques as Direct Injection Mass Spectrometry (DIMS) and Proton Transfer Reaction Mass Spectrometry (PTR-MS) can be used in combination with chemometrics, to determine the suitability of these techniques in authenticity determination. For spice authentication, we developed methods based on these mass spectrometry techniques for the identification of geographical origin. Fifty samples of widely traded spices were analysed: black pepper, chili paprika, nutmeg, vanilla and saffron. Chemometric analysis was initiated with customized automatic m/z variable selection for both DIMS and PTR-MS data. Subsequent exploratory analysis was performed by principal component analysis (PCA), in which maximum of variation between the selected variables was expressed. From this PCA, it was possible to distinguish the different spices and to discriminate between authentic and false material. Partial least square discriminant analysis (PLS-DA) on cross-validated data was conducted and provide a good classification rate with an average of 95.3% of efficiency. Unique about this development is that one method for a whole database seems suitable, rather than one model per spice. This paves the way for uniform herb and spice authentication methods in the future.

Biography:

Wang is the Director of Proteomics and Associate Professor of Biochemistry and Molecular Biology at Indiana University School of Medicine. He received his PhD in Bio-organic Chemistry from Washington University in St. Louis, Missouri, USA and was an NIH NRSA postdoctoral fellow studying mechanism of DNA repair in mammalian system. He has published more than 80 peer-reviewed articles in biochemistry and proteomics related journals. His own research involves studies of cancer drug resistance mechanism and metabolism of omega-3 fatty acid in prostate cancer. Dr. Wang was a recipient of the HUPO (Human Proteome Organization) 2004 Young Investigator Award.

Abstract:

Prostate cancer (PCa) is one of the most common cancers in men. In most PCa cases, tumors progress very slowly but in a small portion of patients, PCa develops into aggressive stages and becomes lethal. A line of evidence has suggested that early inference and dietary prevention are beneficial in PCa patient care. Fish oil (FO), which contains mostly omega-3 fatty acid (n-3 FA), is one of the most widely studied candidate supplements for PCa prevention; however, the molecular mechanism of its function remains elusive. The aim of this study was to identify and characterize factors and pathways involved in fatty acid metabolism in prostate cancer cells through proteomic and phosphoproteomic analyses. Our data suggest that the effects of fatty acids on prostate cancer may be a multi-stage event and autophagy, a double-edged sword, may play a critical role. Global discovery also found other proteins such as non-specific lipid-transfer protein (SCP2), fascin, integrin beta-1, calnexin, and prostate-associated microseminoprotein (MSMP) being altered in fatty acid treated cells. Cell signal pathway analysis reveals that FO modulates some pathways associated with cell cycle and glycolysis. Additionally, the global phosphoproteome study showed different phosphorylation patterns under different FA treatments. Overall, this study shows that fatty acids suppress fatty acid synthase (FASN) activity but only omega-3 fatty acid induces cell death. More functional validation studies are in progress.

Biography:

Yannick Weesepoel finished his PhD in Phytonutrient chemistry with Dr. Jean-Paul Vincken and Prof. Harry Gruppen in September 2014 at the Laboratory of Food Chemistry at Wageningen UR, The Netherlands. In June 2014 he continued his career as a Scientist in Food Authenticity with the group of Prof. Saskia van Ruth at RIKILT, institute for food safety at Wageningen UR. His specialization lies within mass spectrometry and small molecules, chemometrics and portable equipment for food fraud deterrence applications.

Abstract:

Consumer demands are increasing for consumables either produced in an organic way, obtained via fair-trade systems or speciality produced. Especially in the case of coffee these factors seem to have established a permanent role in the choice of consumers for certain types of coffee. In conjunction with higher levels of organic or fair-trade certification and uniqueness of the coffee, trading prices increase inevitably. The (standard) high-end Kopi Luwak coffee can reach up prices to USD 600 per kilo or USD 275 per lbs. As a direct consequence of increased pricing, coffees may be subjected to economically motivated adulteration, e.g. (part of) the coffee is replaced by cheaper counterparts, while package statements may mislead consumers. As a possible solution specialty coffees can be investigated chemically, however no base-line information on how to discriminate these coffees was available. Therefore, 110 market coffees with either a specialty or regular production process were characterized on the basis of their volatile intrinsic markers by high sensitivity proton transfer reaction mass spectrometry (HS PTR-MS). Espresso coffees, Kopi Luwak coffee and organic coffees, could be distinguished by their profiles of volatile compounds with the help of chemometrics. A PLS-DA classification model was used to classify the organic and regular coffees by their HS PTR-MS mass spectra. Extensive cross validation showed correct prediction of 42 out of the 43 (98%) organic coffee samples and 63 out of the 67 (95%) regular coffee samples. It was concluded that the usage of direct headspace analysis techniques like HS PTR-MS is a promising approach to rapid organic and specialty coffee authentication.

Biography:

Abstract:

Vitamin D deficiency is a widespread clinical problem and has been associated with many adverse health outcomes. Analysis of Vitamin D2 (ergocalciferol) and D3 (cholecalciferol) and their major metabolites 25(OH)D2 and 25(OH)D3 has become a high priority topic in clinical analysis. Currently a variety of LC/MS methods have been developed to support vitamin D analysis. These LC/MS methods utilize different transitions, ionization modes, sample preparation strategies, mobile phases and columns. In LC/MS analysis of 25 OH Vitamin D, dehydration (water loss) is the major side reaction. Comparing acetonitrile to methanol, which are typically used as mobile phases for LC separation, acetonitrile does not support hydrogen bond formation; therefore, proton-induced water elimination in-source becomes a major side-reaction, especially given the low pH of the mobile phase and positive mode electrospray and APCI ionization. MeOH, in contrast, supports hydrogen bond formation with the 25(OH)D2 and 25(OH)D3 hydroxyl groups. This efficiently “shields” most of hydroxyl groups by hydrogen bonding, and protects against protonation and resultant water elimination. We found that quantitation of the 25(OH)D from its [M+H]+, “intact” precursor ion, is temperature invariant. In contrast, quantitation using the in-source dehydrated precursor (parent) ion, leads to increased sensitivity with a rise in temperature, due to its better ionization efficiency at higher temperatures. Since actual temperature of droplets can vary with mass spectrometer hardware, flow rate, and mobile phase composition, fluctuations of these factors may contribute additional variability to the assay.

Biography:

Michał Dadlez has completed his PhD in 1992 from the Institute of Biochemistry and Biophysics, Pol. Acad. Sci. and postdoctoral studies from Whitehead Institute, MIT, Cambridge, MA, supervised by Prof. Peter S. Kim. He organized Mass Spectrometry Lab, IBB PAS in 2001, being its head since then. In 2006 he became a Professor of Biophysics. He has published more than 90 papers in peer-reviewed journals. His major field of interest are protein structure studies with use of MS-based methods (HDex, X-linking, Ion Mobility separation, etc.)

Abstract:

The application of mass spectrometry for protein studies extends far beyond classic proteomic analyses in which proteins and their posttranslational modifications are identified and/or quantified. Recent developments of new approaches, like ion mobility separation - IM (providing access to collisional cross section values) or technical improvements in the methods already known (like measurements of the hydrogen-deuterium exchange kinetics) enable efficient application of MS for protein structure studies. These methods allow structural insight into a large group of targets difficult in structure analysis. Among them are oligomerising peptides, including A peptide, main neurotoxic agent in Alzheimer’ s disease, responsible for synaptic dysfunction and neuronal injury. The mechanism of the Aβ peptide self-assembly is still under debate. Using Ion Mobility separation coupled with MS we have measured collisional cross-section values of different oligomeric forms of A, from dimers to hexadecamers. For several oligomers, at least two different forms of different Ω values were detected, indicating the presence of at least two families of conformers: compact and extended. We have also characterized numerous factors shaping the compact/extended species equilibria, like metal binding or point mutations. IM-MS thus detected oligomeric species being both on-pathway in the process of fibril formation, but also alternative structures which may represent potentially most neurotoxic, off-pathway oligomers and allowed their basic structural characterisation in the context of complex mixture of interconverting species.

Biography:

Purushottam Chakraborty is a senior professor at Saha Institute of Nuclear Physics, Kolkata, India and an honorary professor at the University of Pretoria, South Africa. His research interests range from ‘ion beam analysis of materials’ to ‘x-uv optics and photonics’. He worked at many renowned centres like FOM – Institute for Atomic and Molecular Physics, Padova University, ICTP, Laval University, Osaka Electro-communication University, etc. He has published more than 125 papers including reviews and book-chapters. He has been awarded the ‘most eminent mass spectrometrist of India’ and is one of the world leaders in Secondary Ion Mass Spectrometry (SIMS).

Abstract:

Excellent detection sensitivity, high dynamic range and superior depth resolution make the SIMS technique extremely powerful for the analysis of low-dimensional structures. However, a serious problem in SIMS analysis lies in its ‘‘matrix effect’’ that hinders the materials quantification. Appropriate corrective measures are therefore, needed to calibrate the secondary ion currents into respective concentrations for accurate compositional analysis. Working in the MCs+-SIMS mode (M – element to be analyzed, Cs+ – bombarding ions) can circumvent the matrix effect. The emission process for the neutral species M0 is decoupled from the MCs+ ion formation process, in analogy with the ion formation in secondary neutral mass spectrometry (SNMS), resulting in a drastic decrease in matrix effect in the MCs+-SIMS mode. Although this technique has found its applicability in direct quantification, it generally suffers from a low useful yield. In such cases, detection of MCsn+ (n = 2,3,. . .) molecular ions offers a better sensitivity, even by several orders of magnitude. A complete understanding on the formation mechanisms of these MCsn+ (n = 2,3,. . .) molecular ion complexes formed in the SIMS process has been explored. We have demonstrated the prospective use of this innovative MCsn+-SIMS technique in direct interfacial analysis of ultra-thin films, metallic multilayers, semiconductor superlattices, quantum well structures and compositional analysis of MBE grown Si1-xGex alloys without ‘standards’. The talk will address on the fundamentals, challenges and applications of the novel MCsn+-SIMS technique in all its complexities.

Biography:

Rahmatollah Rahimi has completed his PhD at Howard University (USA). He has been now serving as a full Professor in the Department of Chemistry, Iran University of Science and Technology. He has published about 90 papers in reputed journals

Abstract:

Sulfur/graphene catalyst was invented to be recyclable and impressive for selective oxidation of benzyl alcohol under simple reaction conditions.In this study, graphene oxide (GO) was prepared by a Hummers and Offman method. A one-step environmentally way is developed to prepare sulfur/reduced graphene. Sulfur/reduced graphene oxide (S/RGO) composite has been synthesized using a simple and low cost method with thiosulfate and graphene oxide (GO) as precursors. The powder was determined by X-ray diffraction, Fourier transform infrared spectroscopy and scanning electron microscopy images. After that, As-prepared sulfur graphene has been used for oxidation of benzyl alcohols to benzaldehyde. Acetonitrile has been used as solvent in the reaction. This composite were able to oxidation of benzyl alcohol to benzaldehyde with high efficiency (about 96%). This conversion has been demonstrated by gas chromatography equipped with mass spectroscopy. Substituted benzyl alcohols also can be oxidized by O2 to corresponding aldehydes with high selectivity in presence of catalyst. However this conversion has been reported difference yield with graphene oxide and another composite of graphene, but the first time was reported sulfur/graphene.

Biography:

Dr. Rahmatollah Rahimi has completed his PhD at Howard University (USA). He has been now serving as a full Professor in the Department of Chemistry, Iran University of Science and Technology. He has published about 90 papers in reputed journals

Abstract:

Disulfides are the key component of organic and bioorganic process such as disulfide bond formation on peptides, control cellular redox potential in biological systems and they are used as protecting agent of thiols under oxidative condition. The most important method of producing disulfides is the oxidation of thiols. A board range of reagent and catalysts are used for oxidation of thiols to disulfides like molecular oxygen, metal oxides, nitric oxides, halogens, sodium perborate and DMSO. Tin oxide is a metal oxide that is wildly used in different area like electrode material, a transparent electrode, glass sensing electrode and they are used as hydrogen, carbon monoxide, hydrocarbon and alcohol sensor. In this work SnO2 nanostructures were synthesized by simple precipitation method by adding ammonia directly to SnCl2 solution and it was used as catalyst for oxidation of thiols derivatives to disulfides. It was a selective and highly efficient catalyst. SnO2 nanostructures were characterized by XRD and SEM and products of thiol oxidation were identified by GC-mass spectroscopy.

Biography:

Dr. Rahmatollah Rahimi has completed his PhD at Howard University (USA). He has been now serving as a full Professor in the Department of Chemistry, Iran University of Science and Technology. He has published about 90 papers in reputed journals

Abstract:

In this study, ZnO nanoparticles have been immobilized on the surface of Zn-Fe2O4 nanohallow spheres by sol-gel method and the core-shell structure of Zn-Fe2O4@ZnO nanohallow spheres were characterization by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and vibration sample magnetometery (VSM). The SEM images reveal formation of sphere structure. The TEM micrographs confirm that the size of catalyst is about 150 nm and the thickness of ZnO shell is smaller than 30 nm. Results of VSM illustrate that Zn-Fe2O4@ZnOparticles are superparamagnetic. The catalytic activity of Zn-Fe2O4@ZnO nanohallow spheres was investigated for oxidation of benzylic alcohols using H2O2 as an oxidant. The resultant Zn-Fe2O4@ZnO nanohallow spheres were highly selective for oxidation of benzyl alcohols to corresponding benzaldehydes. It was found that the as–prepared catalyst display the excellent performance, whereby, 100% selectivity of benzaldehyde was achieved at close to 92% conversion.

Biography:

Abstract:

Surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) using nanomaterials as matrix has become a powerful technique for the analysis of biomolecules, including proteins, DNA, peptides, aminothiols, and carbohydrates. SALDI-MS has been developed to improve sensitivity and quantitation, with minimum problems associated with sweet spots. Several successful examples of the analysis of biomolecules through SALDI-MS will be discussed in my talk. SALDI-MS with HgTe nanostructures as matrix is useful for the detection of proteins such as IgG, protein G, and their complexes. SALDI-MS allows simultaneous detection of single- and double-stranded oligodeoxynucleotides, with limits of detection at the femtomole-to-picomole level and sample-to-sample intensity variation < 23%. SALDI-MS using gold nanoparticles (Au NPs) is sensitive for the detection of the digest of Cyt c in the HeLa cells treated with etoposide (a commercial drug) or carbon dots (potential drug).

Biography:

Erik Dümichen is a PhD student at the Federal Institute for Materials Research and Testing (BAM) in Germany since 2012. He is a young scientist with experience in investigation of polymers by thermo-analytical and chromatographic methods. He developed a robust and easy to handle method for the identification and quantification of thermal- and thermo-oxidative degradation products.

Abstract:

The common analyzing techniques for polymer degradation mechanism are thermogravimetry (TGA) coupled to Fourier transformed infrared spectroscopy or mass spectrometry (TGA-FTIR/MS) and pyrolysis gas chromatography mass spectrometry (Py-GC-MS). TGA-FTIR and TGA-MS are powerful tools for the analysis of decomposition molecules like water, methane, carbon dioxide or ammonia due the specific signals of these small molecules with high symmetry. With Py-GC-MS the hydrocarbon decomposition products can be separated by a chromatographic column and identified over mass spectrometry. Therefore, an unambiguous identification of characteristic polymeric decomposition product is possible. However, using Py-GC-MS only small sample amounts can be analyzed (0,1 – 0,5 mg) and small transfer capillaries are used which cause problems through contamination or even blockage. Therefore, a quantification or semi-quantification of decomposition products is difficult to do. A new technique was established that is called thermal extraction and desorption gas chromatography mass spectrometry (TED-GC-MS). The method combines the advantage of using the thermogravimetry as thermal extractor: mass signal, using inert and oxidizing atmosphere, designed to work under the formation of big amounts of dirt, easy sample preparation, as well as the advantage of a chromatographic system that can separate the degradation products. These decomposition products can be certainly identified with reference mass spectra. The extraction was realized by the adsorption of the degradation products out of the TGA on twister. These twister are small stir bras with an defined amount of adsorption material (PDMS) on the surface. Subsequently these twisters were analyzed by thermal desorption gas chromatography mass spectrometry. The desorption is carried out with ambient temperatures (about 200 °C). Thus, the MS-system stays clean and no problems with contamination or even a blockage were observed. Quantitative as well as semi-quantitative measurements can be carried out due to this defined adsorption capacity. We will present some results about various topics using this new method. Advantages and disadvantages will be presented and a comparison with established methods.

Biography:

Benjamin J. Bythell received his MChem. degree from the University of Bath, UK, in 2002 and Ph.D. from Oregon State University in 2007. He held postdoctoral fellowships at the German Cancer Research Center in Heidelberg (2008-2010) and at the National High Magnetic Field Laboratory at Florida State University (2010-2013). He joined the University of Missouri-St. Louis faculty in 2013. Dr. Bythell works at the interface between analytical, computational and biophysical chemistry where he strives to understand the structure, reactivity and gas-phase behavior of biologically- and industrially-important chemicals.

Abstract:

Fundamentally, chemical structure determines the properties and potential functions of any given molecule. Consequently, to effectively and consistently identify chemicals with tandem mass spectrometry the gas-phase structures occupied by our analyte ions will have direct influence on which fragmentation pathways are populated, and thus, on the resulting mass spectrum. Ideally, we desire the ability to decipher both the elemental composition (CcHhNnOoSsPp) and structural information on unknown compounds. To accomplish this successfully, an understanding of the gas-phase fragmentation chemistries likely to be in play is of substantial benefit. Consequently we need a reasonable means of modelling the processes occurring within the mass spectrometer or a spectral database of all possible compounds under the particular experimental conditions employed. The latter, optimal approach is impractical for many compound types as it requires reference spectra generated from prior confident identification of each possible synthesized/isolated analyte. Thus, we require a robust model for our particular compound class that is flexible and accurate enough to deal with how and why different analyte ions form particular conformations, and what affect this has on their gas-phase fragmentation chemistry and resultant mass spectra. I will discuss recent density functional theory and experimental findings and their implications for structural identification of protonated systems with MS/MS.

Biography:

Wenjie Cao is a Staff Scientist of the Analytical at the SABIC Technology Center at Riyadh. Received his Ph.D. from Professor John Calvin Giddings’ Group at the University of Utah. Ph.D. research was on polymer separation and characterization by Thermal Field-Flow Fractionation. He was working for DuPont as a Research Investigator for four years before joined SABIC in 2012. Contributor to the book of the Encyclopedia of Chromatography. More than eighteen publications and presentations in peer-reviewed scientific journals and international conferences. Filed five patents since joined SABIC in 2012.

Abstract:

For the triple quadrupole LC-MS/MS instrument, the primary purpose or the most significant feature is the highest sensitivity among almost all, if not all, of the LC-MS/MS instruments by doing the Multiple Reaction Monitoring (MRM) testing. The ionization of a chemical in LC-MS/MS could be done with different ionization modes. The common ones are Electron Spray Ionization (ESI), the Atmospheric Pressure Chemical Ionization (APCI), and the Atmospheric Pressure Photo Ionization (APPI). The species and amount of adduct ions produced at each mode are quite different. Some type of the adduct ions may complicate the MRM testing by decreasing the sensitivities while some other adduct ions may prevent any reliable MRM testing. This research is going to do investigation on the differences of the types of adduct ions produced, and the amount of adduct ions produced at different ionization techniques. Some examples will be presented to show how the adduct ions may complicate the MRM testing, and the best ionization mode for some type of the targeted chemicals.

Biography:

Tatsuya Fujino, male, spectroscopy, Dr. of sc. from Tokyo Institute of Technology in 1998. He worked as a postdoctoral researcher of Japan Society for the Promotion of Science (JSPS) at Institute of Molecular Science (IMS) 1998-2002 and as a special postdoctoral researcher at RIKEN 2002-2005. Since 2005-till date, he is an associate professor at Tokyo Metropolitan University. New century award from the Japan Society for Analytical Chemistry (JSAC) in 2006. Best presentation awards from the Spectroscopical Society of Japan (SPSJ) in 2004 and from the Kanto branch of Chemical Society of Japan (CSJ) in 2007.

Abstract:

Nanometer-sized cadmium telluride particles were used as an inorganic matrix for the laser desorption/ionization mass spectrometry of free fatty acids in human serum. By changing the excitation power and the amount of a hole scavenger, the peak of deprotonated fatty acids was investigated. It was understood that the ionization of fatty acids were due to the biexciton Auger recombination and electron ejection from CdTe. CdTe were then loaded on zeolite surface. The peak intensity enhancement of the deprotonated ion of fatty acid were observed. This phenomenon was explained by measuring the carrier lifetime for Auger recombination in CdTe. In addition, reproducibility of fatty acid ions was highly improved reflecting homogeneous distribution of CdTe on zeolite surface. CdTe loaded on a zeolite surface (HM20) was successfully applied to the quantitative analysis of Ste in human serum by isotope dilution using 13C18-Ste. The concentration of Ste in human serum samples was estimated to be 76.62 mg/kg with the standard deviation (SD) of 2.37 mg/kg.

Biography:

R.K. Vatsa completed his Ph.D. from Bombay University, INDIA and did his postdoctoral studies at Heidelberg University, Germany. He is currently heading the Structural Chemistry Section in Chemistry Division of BARC. He has published more than 125 papers in reputed journals. He is vice-president of Indian Society for Mass Spectrometry (ISMAS) and elected fellow of National Academy of Sciences, INDIA.

Abstract:

This presentation will focus on multiple ionization and Coulomb explosion observed in different atomic and molecular cluster systems at laser intensities of 109 W/cm2. Clusters were prepared by supersonic expansion and ionized by Nd:YAG laser (10 ns pulse width). Multiply charged atomic ions possessing high kinetic energies were detected by time-of-flight mass spectrometry. Along with this, presence of energetic electrons (with 25-30 eV kinetic energy) was also confirmed by retarding field method. Formation of such highly charged atomic ions at these intensities is quite unusual and difficult to explain theoretically. In order to understand different factors responsible for enhanced ionization, a systematic study was conducted in which wavelength, cluster size and cluster composition were varied. Ionisation wavelength was found to have a pronounced effect on the charge state of ions and a threshold cluster size is essential. For gigawatt pulses interacting with a cluster, primary step is multiphoton ionization of atoms/molecules giving rise to an ion and electron, the latter then absorbs energy from the optical field. In case of clusters, electrons which have been detached from parent atom, but are trapped inside the cluster are called quasi-free electrons. The results show that presence of quasi-free electrons and their confinement inside the cluster is very crucial for generation of multiply charged ions.

Biography:

Professor Webb completed his PhD from Salford University and performed postdoctoral studies the Naval Postgraduate School in Monterey, Ca, USA. He is now the director of the Surrey Ion Beam Centre, the National Centre for Ion Beam Applications in the UK. He has published more than 250 papers in reputed journals and has served on editorial boards of a number of journals and is a member of the scientific program committees of a number of international conferences.

Abstract:

In the 1970s it was shown that MeV heavy ions efficiently sputter insulating targets. An analysis technique based on this, Plasma Desorption Mass Spectrometry (PDMS), employed Cf fission fragments as a source of MeV heavy ions. The technique was able to desorb large molecular secondary ions (>10kDa) from solid surfaces, which at the time was not possible with other mass spectrometry techniques. In the 1980s, however, Laser Desorption techniques (such as MALDI) were also being demonstrated, which didn’t require the presence of radioactive material or an accelerator. Consequently PDMS was all but forgotten. There has been a resurgence of interest in PDMS (renamed MeV-SIMS) when it was shown that a focused ion beam could be used to produce images with a much higher spatial resolution than is currently possible with e.g. MALDI. It was also been demonstrated that the technique can be performed simultaneously with PIXE measurements. One of the limitations of ionization techniques is the effects of the matrix on the secondary ion yield which can make even relative measurements difficult. The use of the elemental signals removes some ambiguity. New equipment being commissioned will be described which will allow simultaneous MeV-SIMS and PIXE to be collected in full ambient pressures with a micron beam resolution. Spectra and images taken for the first time in this scheme at fully ambient pressure are presented to demonstrate the potential of this new instrument.

Biography:

Helmut Schwarz is chief Scientist in Technische Universität Berlin, Germany. He has worked on detailed insight in the active site of catalysts is provided and – in combination with computational chemistry – mechanistic aspects of as well as the elementary steps involved in the making and breaking of chemical bonds. He has published more than 300 research papers.

Abstract:

We shall present selected examples of gas-phase reactions which are of timely interest for the catalytic activation of small molecules. Due to the very nature of the experiments, detailed insight in the active site of catalysts is provided and – in combination with computational chemistry – mechanistic aspects of as well as the elementary steps involved in the making and breaking of chemical bonds are revealed. Examples to be discussed include inter alia: (i) Metal-mediated carbon-carbon bond formation; (ii) low-temperature, catalytic oxidation of CO; (iii) oxygen-centered radicals as active sites in catalytic hydrocarbon activation, e.g. the oxidative coupling of methane 2CH4  C2H6. It will be shown that mass-spectrometry based studies on 'isolated' reactive species provide an ideal arena for probing experimentally the energetics and kinetics of a chemical reaction in an unperturbed environment at a strictly molecular level, and thus enable the characterization of crucial intermediates that have previously not been within the reach of conventional condensed-phase techniques.

Biography:

Athula B. Attygalle is associate professor in Stevens Institute of Technology, USA .After an international scientific safari (University of Ceylon, University of Sri Lanka, Tokyo Institute of Technology, University of Keele, University of Erlangen-Nürnberg, and University of Houston), Attygalle became the Director of the mass spectrometry facility of Cornell University in 1991, After 12 years at Cornell ten years, Attygalle became a Professor at Stevens Institute of Technology in Hoboken, NJ, in 2001. Attygalle has published over 200 papers in reputed journals and holds five U.S. patents.

Abstract:

Some ambient-ionization techniques have revolutionized direct sample analysis by mass spectrometry because the need of a solvent spray can be eliminated. Recently described Helium-Plasma Ionization Mass Spectrometry (HePI) has been recognized as one of the promising novel procedures for ionizing a variety of organic and inorganic compounds. Under positive-ion generating HePI conditions some molecules undergo protonation [(M + H)+] while others generate molecular ions (M+•). Our recent results show that the formation of molecular ions or protonated molecules can be swung back-and forth in a controlled manner. By manipulating the atmosphere surrounding the source, we can generate molecular ions from aromatic hydrocarbons, and virtually eliminate the protonated species. The ionization is effected by a direct Penning mechanism due to interactions of the gas-phase analyte with metastable helium atoms. However, when the ambient gases bear even traces of moisture, the analytes are ionized by proton transfer reactions with gaseous H3O+. We demonstrate that by controlling experimental conditions, the abundance of molecular ions and protonated molecules in a HePI source can be manipulated.

Biography:

Chrys Wesdemiotis received his Ph.D. degree with Helmut Schwarz at Technische Universität Berlin in 1979. He was a postdoctoral fellow with Fred W. McLafferty at Cornell University in 1980. After completing his military service in Greece (1981-3), he returned to Cornell as a senior research associate (1983-9). In 1989, he joined the faculty of the University of Akron, where he currently is Distinguished Professor of Chemistry, Polymers Science, and Integrated Bioscience. Research in the Wesdemiotis group focuses on the development and applications of multi-dimensional mass spectrometry methods for the characterization of new synthetic polymers and polymer-biomolecule conjugates and interfaces. Wesdemiotis served as a Member-at-Large for Education in the Board of Directors of the American Society for Mass Spectrometry (ASMS) and is a Fellow of the American Association for the Advancement of Science (AAAS). He is an Editor of the European Journal of Mass Spectrometry and a member of the Editorial Boards of the Journal of the American Society for Mass Spectrometry, International Journal of Mass Spectrometry, and Mass Spectrometry Reviews.

Abstract:

Matrix-assisted laser desorption ionization and electrospray ionization have enabled mass spectrometry (MS) analyses for a wide variety of synthetic polymers, but considerable challenges still exist. Polymerizations often create complex mixtures that are impossible to characterize by single-stage MS because of superimposed compositions or discrimination effects in the ionization or detection steps. These problems can partly be resolved by tandem and multistage mass spectrometry (MS2); depending on the polymer, MS2 may lead to fragmentation patterns that permit unequivocal identification of the corresponding architecture and comonomer sequence. For the characterization of more complex systems, MS must be combined with separation methods, such as liquid chromatography (LC) or ion mobility (IM) spectrometry. Particularly powerful for such studies are approaches that couple ultrahigh performance LC (UPLC), which significantly reduces elution times, with the ion mobility dimension, which enhances separation and spectral decongestion. Different combinations of LC, IM, and MS2 can be used to address questions about polymer composition, structure, and 3D architecture. Analyses involving top-down methods (the entire analysis is performed in the mass spectrometer) as well as methods including prior LC separation or mild, in situ thermal desorption/degradation will be demonstrated with a variety of synthetic polymers, including new amphiphilic copolymers, hybrid materials containing bioactive peptides, and supramolecular systems.

Biography:

Rafael Garrett received his BSc in Pharmacy from Fluminense Federal University (UFF), Brazil; MSc in Chemistry of Natural Products from Federal University of Rio de Janeiro (NPPN-UFRJ); and PhD in Chemistry at the prestigious Chemistry Institute of UFRJ under the supervision of Profs. Marcos Eberlin and Claudia Rezende, with an internship at Prof. Ifa’s lab at York University (Canada), working with modern mass spectrometry approaches to study coffee. He is a pioneer in the use of high-resolution MS and Ambient Ionization techniques for coffee analysis. Currently, he works as postdoctoral fellow at UFRJ.

Abstract:

Coffee is one of the most consumed beverage and an important commodity for many developing countries. Its quality is generally related to good agriculture and handling practices. Geographic origin also plays a central role and discussions about terroir are common nowadays. Chemical investigations of coffees usually deal with low number of analytes using elaborated or time-consuming protocols and do not allow high-throughput analysis. The development of new, simple and fast methods for coffee analysis and quality control is still required. Ambient ionization MS comprises a set of techniques where the ionization process occurs outside the mass spectrometer, in open-air environment. Generally, they are simple and provide direct analyses. Here, ambient ionization techniques (Paper Spray, EASI and DESI-Imaging) are described to study and discriminate coffees. Coffee Spray ionization, where a bean slice is used as both sample and substrate to perform electrospray, is also introduced. Paper Spray fingerprinting combined with multivariate statistics was successfully employed as a fast and simple way to discriminate coffee origin. EASI-MS was used to study intact coffee beans submitted to different post-harvest treatments and wax components related to stomach irritations were easily detected. DESI-Imaging of a cross-sectioned bean revealed different spatial distribution of important phenolic compounds present in coffee. All these ambient ionization techniques proved to be very useful in coffee science. They can be employed to study a single bean or for high-throughput analysis, while preserving its integrity. Besides, with the improvements of portable MS, they have potential to be used in field studies.

Biography:

Professor Auray-Blais is the Director of the Quebec Mass Neonatal Urinary Screening Program for hereditary metabolic disorders. She holds a Ph.D. in radiobiology from the Université de Sherbrooke and postdoctoral studies from Duke University Medical Center, NC. She has a master’s degree in Health Law from the Faculty of Law at the Université de Sherbrooke. She is the author of 200 publications, abstracts and articles. She is a professor in the Medical Genetics Division in the Pediatrics department at the Faculty of Medicine and Health Sciences in Sherbrooke. She is the Scientific Director for the Waters-CHUS Expertise Centre in Clinical Mass Spectrometry.

Abstract:

In order to increase the number of treatable disorders screened by the Mass Urinary Screening Program in the Province of Quebec, we developed and validated a tandem mass spectrometry multiplex method for creatine synthesis and transport disorders, Triple H syndrome and ornithine transcarbamylase deficiency. We targeted specific-related biomarkers such as creatine, guanidineacetate, uracil, orotic acid and creatinine using urine collected on filter paper. This method showed good linearity with a mean coefficient of regression at 0.9985 for all metabolites, as well as precision and accuracy for the intraday and interday assays at 15%. All abnormal patients were well discriminated from controls. We have shown that this methodology is feasible for mass or high-risk screening with high sensitivity and reproducibility. Regarding lysosomal storage disorders, we have performed a time of flight mass spectrometry metabolomic study for biomarker discovery for Fabry disease, a multisystemic, X-linked complex disorder. We detected novel analogues of a glycosphingolipid, globotriaosylsphingosine (lyso-Gb3), which were found to be increased in Fabry patients presenting the cardiac variant mutation of the disease. Robust tandem mass spectrometry methods were devised and validated for both urine and plasma. We have also devised a UPLC-MS/MS method for methylated and non-methylated forms of globoriaosylceramide (Gb3), another biomarker detected in Fabry disease patients. These latter methods allowed the quantification of biomarkers in biological fluids of affected Fabry patients and controls, and are aimed at the evaluation of correlations to determine disease-severity and progression.

Biography:

Sermin Tetik has completed his PhD at the 2003 years from University of Marmara (Istanbul-Turkey). She is a academician at the same University in Istanbul-Turkey as a associated professor and project director of a research team focusing on thrombosis -hemostasis area and she completed before 6 months in Cyprus as a foundar ( vice-Dean) of a new Pharmacy Faculty at University of International Cyprus . She has published more than 20 papers and 50 international abstract in reputed journals and conferences, serving as an editorial board member of repute.

Abstract:

Mass Spectrometry (MS) has become the method of fields and applications, including proteomics and proteomic analysis. MS is now routinely used to identify and quantify known and unknown proteins in our labs as a fundamental tool. Its accuracy, flexibility and sensitivity have permissed new approachs in the biological area, characterization of biopharmaceutics and diagnostic criteria. MS has allowed it possible to analyze with of all is many types of mass analyzers including ESI, MALDI, FT-MS, ion trap, time-of-flight, quadrupole, etc. Mass analysis of protein and proteomic(s) products have ranged in mass between 50-300.000 daltons, in attomole through nanomole quantities. Sample preparation techniques for MS-analysis is a important step in the proteomics workflow. In the protein analysis sample preparation is the most variable and time consuming step. In proteomics laboratory sample preparation, instrumentation and software are all critical to success and high quality results. There is different sample preparation techniques because we have no standard method for preparing including type, complexity, source etc. Proper sample preparation means better results.

Biography:

Shubhen Kapila is the MSMC Endowed Professor at the Missouri University of Science & Technology – a campus of the University of Missouri. He obtained his Ph.D. in Chemistry from Dalhousie University, Nova Scotia, Canada and joined the University of Missouri System in 1977. He has published over 165 papers in reputed journals and has supervised research of more than 100 graduate students and post-doctoral fellows. He has been involved with national and international standardization bodies ASTM, ANSI and International Electrotechnical Commission (IEC), he has served as the convener of standard setting working groups and project teams.

Abstract:

There is a need for rapid determination of additives, contaminants and by-products in insulating liquids. Desorption electrospray ionization (DESI) has emerged as technique of choice for a variety of applications. Use of DESI for characterization and quantification of additives, contaminants and by-products in insulating liquids has been explored in our laboratory. Results obtained for selected target analytes were compared with the results obtained with the traditional methodologies based on high pressure liquid chromatography electrospray ionization mass spectrometry (HPLC-ESI MS), gas chromatography – mass spectrometry (GC-MS) and Fourier transform infrared (FTIR) spectrometry. Results showed that DESI offers a very rapid of quantitative determinations of selected analytes in complex matrices such as the insulating liquids. The most attractive feature of the technique is no or minimal sample pre-treatment thus permitting very rapid analysis. However, the precision obtained with the technique was found to be inferior to that obtained the other technique.

Biography:

Abstract:

The Metabolic Syndrome (MeS) is a clustering of components that reflect over nutrition, sedentary lifestyles and resultant obesity. MeS includes the clustering of abdominal obesity, insulin resistance, dyslipidemia and elevated blood pressure1. The prevalence of the MeS is globally increasing to epidemic proportions and is associated with an approximate doubling of the cardiovascular disease risk and a 5-fold increased risk for type 2 diabetes. Observational studies suggest that dairy foods consumption may prevent the development of MeS and its related disorders2; both whey and casein proteins stimulate insulin secretion and have the potential to improve tissue glucose uptake and suppress postprandial blood glucose3. The milk protein effect on postprandial lipaemia and chronic dyslipidaemia is not well defined as studies have produced different findings4. Although some studies show beneficial effects of milk fat especially rich in short- and medium-chain fatty acids on weight control, glucose and lipid metabolism5, 6, results are still conflicting. To gain further insight into the nutritional impact of milk protein and milk fat metabolomics is proposed as a pioneer approach. It may help to increase our understanding of the influence of milk components e.g. on MeS by the advent of comprehensive analysis of low-molecular-weight compounds (metabolites) present in complex samples such as human biofluids collected from an intervention study. Liquid chormoatography- and gas chromatography-quadrupole time of flight mass spectrometry (LC-MS and GC-MS Q-ToF) untargeted metabolomics were applied to develop multivariate models to identify biomarkers related to the intake of milk lipids with high or low content of medium-chain saturated fatty acids (MCFA), and milk proteins (whey vs. caseins) on metabolic profile in 52 subjects with abdominal obesity. Serum (fasted and 240 min after meal) and urine (fasted and 360 min after meal) samples were collected before and after the 12-weeks randomized, controlled, parallel, iso-caloric dietary intervention. Participants were randomized to one of four groups according to the intervention diet; low amount of MCFA (L-MCFA) + Whey, high amount of MCFA (H-MCFA) + Whey, L-MCFA + Casein, H-MCFA + Casein. Using LC-MS Q-ToF on blood samples yield both post prandial and long term effects on the metabolic profile of blood. A clear separation in the metabolic profile was observed for the group with H-MCFA + Casein compared to the three other groups while no effect on the metabolomics profile in urine was observed. Using GC-MS Q-ToF we have found a separation of all four groups when analysing the long term effect on the metabolic profile in urine. By combining LC-MS Q-ToF with GC-MS Q-ToF data thus secured optimal analysis of more metabolites, which now are being identified and related to the intake of dairy products and their effect on obese individuals. References

Biography:

Paul Wenthold received a PhD from Purdue University, and was a postdoctoral associate at the University of Colorado. After a short appointment as an assistant professor at Texas Tech University, he moved back to Purdue, where he was promoted to associate professor in 2004. His work involves the investigation of physical organic chemistry by using mass spectrometry, and is interested in the electronic structures of open-shell intermediates and the reactivity and thermochemical properties of reactive molecules in the gas phase.

Abstract:

The decomposition of canonical structures of α-amino acids has been investigated by using charge remote fragmentation. Neutral amino acids in condensed-phase have zwitterionic structures and are non-volatile, which makes gas-phase studies difficult. Previously reported mass spec studies of amino acids generally utilize ionization (protonation, deprotonation, metallation) of the amino acid group. In this work, we describe studies of amino acids where the ionization occurs away from the α-amino acid moiety. Therefore, we report studies of para-substituted phenylalanine derivatives, where the substituents are either a sulfonate (SO3) or trimethylammonium (NMe3+) group. For both ions, we find that the main dissociation pathway involves loss of ammonia to form an α-lactone. For the sulfonate, loss of ammonia is the only dissociation pathway, whereas loss of CO2 is also observed for the ammonium derivative, indicating an effect of charge polarity on the dissociation. The dissociation processes of the α-lactones have also been examined, and are also found to be dependent on the charge polarity, which is readily explained in terms of benzylic positive charge character in the decomposition transition state. We have also examined the effect of solvation on the dissociation, and found that while addition of a single water does not change the reaction, dramatic changes are observed upon addition of two solvent molecules, which indicates interaction between the solvent and the amino acid group.

Biography:

Abstract:

Protein biomarkers are important for early detection, prognosis, and drug response monitoring in cancer treatment. Plasma-based protein markers were sought after because of the easiness of sample collection. However, the potential useful biomarkers are usually masked by the top abundant proteins such as serum albumin and immunoglobulins. In this study, we used sequential window acquisition of all theoretical MS (SWATH) approach to perform non-labelled semi-quantitative proteomics discovery of proximal fluid of four groups in totally 14 liver cancer xenograft models. The tumor tissues were isolated from the mouse host and then resuspended in buffer saline briefly to allow the tissue to secret enough proteins for detection. The secretory proteins in the buffer was digested with trypsin and were fed into Eksigent nanoLC cHiP based C18 column and then identified with ABSCIEX TripleTOF5600 mass spectrometer. A total of 41 runs were performed and each sample was run in duplicates or triplicates. More than 1600 peptides were identified. Principal component analysis was able to segregate the samples into four groups. Significant analysis of the peptides found that 134 peptides were the minimal set of signatures for clear classification of the four groups of samples with highest specificity (1) and sensitivity (1). The product ion intensities provided a hint for development of multiple reaction monitoring (MRM) assay for absolute quantification of target peptides in the future. In summary, SWATH approach of protein marker discovery can lead not only to deep discovery of protein markers but also provide a lead to target peptide selection for the next phase absolute quantification using MRM assays.

Biography:

Sermin Tetik has completed his PhD at the 2003 years from University of Marmara (Istanbul-Turkey). She is a academician at the same University in Istanbul-Turkey as a associated professor and project director of a research team focusing on thrombosis -hemostasis area and she completed a different duty before 6 months in North Cyprus as a foundar ( vice-Dean) of a new Pharmacy Faculty at University of International Cyprus. She has published more than 20 papers and 50 international abstract in reputed journals and conferences, serving as an editorial board member of repute.

Abstract:

Mass Spectrometry (MS) has become the method of fields and applications, including proteomics and proteomic analysis. MS is now routinely used to identify and quantify known and unknown proteins in our labs as a fundamental tool. Its accuracy, flexibility and sensitivity have permissed new approachs in the biological area, characterization of biopharmaceutics and diagnostic criteria. The utility of MS for simple and complex protein and peptide mixtures analyses gave us the ability to develop such accurate results within the last two decades. The development of high-throughput and quantitative MS proteomics workflows has supported to scope of what we know about protein structure, function, modification and global protein dynamics. This overview outlines the role of mass spectrometry in the field of proteomics, MS methodology and instrumentation.

Biography:

Friso van Amerom holds a PhD in physical chemistry from the University of Amsterdam and University Leiden (Netherlands). He is a research scientist at SRI International and Managing Director of Mini-Mass Consulting, Inc., delivering services and expertise to clients in the field of mass spectrometry. He reviews scientific instrumentation for customers and works with NASA, focusing on building and testing mass spectrometers for planetary science

Abstract:

Miniaturization of mass spectrometers opens up a wide range of new applications that require portability during chemical measurements. Mass production of micro-sized mass spectrometers ultimately reduces production costs, allowing miniaturized spectrometers to be used as point-of-care medical diagnostics instruments. A new fabrication method, simulations, and experimental results for micromachined cylindrical ion trap (μ-CIT) arrays for use in miniaturized mass spectrometers is therefore presented. Simulations were performed in SIMION 7.0 to determine the optimum range of μ-CIT z0/r0 to be fabricated and tested. Micromachined μ-CIT arrays were fabricated in a silicon-on-insulator substrate. A series of z0/r0 were chosen in incremental steps of 3% for each array by changing r0 from 308 to 392 μm, while keeping z0 fixed at 355 μm, resulting in a range of z0/r0 from 1.16 to 0.92 (nine geometries in total). This resulted in fast, iterative measurements of the differences in the mass spectra from μ-CITs with different ratios of half-axial to half-radial dimensions (z0/r0). A deep reactive ion etching technique was used to create the cylindrical structures. Surface metallization created the ion trap electrodes. Symmetrical arrays of half-CITs were fabricated, diced, and bonded back-to-back to obtain complete μ-CIT array chips. Mass spectra were obtained experimentally from each trap geometry, and μ-CIT performance was found to follow the trend with respect to z0/r0 observed in the simulations. Axial modulation on one endplate electrode resulted in mass spectra with full-width-at-half-maximum peaks of 0.4 amu.

Biography:

Rahmatollah Rahimi has completed his PhD at Howard University (USA). He has been now serving as a full Professor in the Department of Chemistry, Iran University of Science and Technology. He has published about 90 papers in reputed journals

Abstract:

Metal/Oxide nanoparticles due to their special properties such as stability, easy synthesis and reusability are interesting as catalyst for organic oxidation reaction. Ni/Bi2WO6 nanoparticles were synthesized by simple hydrothermal method. The obtained-products were characterized by EDS, X-ray Diffraction (XRD) and other techniques. The catalytic activity of Ni/Bi2WO6 nanoparticles on oxidation of different alcohols with hydrogen peroxide as green oxidant was investigated. Benzyl alcohol has been used as model alcohol to examine for oxidative activity and selectivity at reflux conditions. Yield of reaction were determined by GC-Mass instrument.

Biography:

Petr Vaňhara (1980) finished his PhD in molecular and cell biology in 2009 at Masaryk University, Czech Republic. He has an international experience from laboratories in Germany, France and Austria. Since 2005 he works in biomedical research. In 2011 he joined the Faculty of Medicine, Masaryk University as the assistant professor, researcher and project leader. He is focused on biomedical applications of pluripotent stem cells, e.g. tissue regeneration and engineering, cancer modeling or biosafety issues related to the genetic and phenotypic stability, including development of novel bioanalytical phenotyping techniques. He published more than 20 internationally recognized papers in reputed journals.

Abstract:

Current biomedical research is highly dependent on correct use of cell lines in vitro. There is a constant risk of cell misidentification, cross-contamination or unwanted phenotypic or genetic and epigenetics shifts in in vitro cultures. In human cells, the current golden standard is identification of short tandem repeats (STR) profile in the genome and their comparison with a database of known profiles. This approach can easily distinguish cell lines derived from different individuals, however, in is limited in case of non-human species without a defined database, in case of clones or different cells from the same individual or in case of phenotypic alterations that does not change the STR profile. To obtain DNA-independent authentication tool, we developed, optimized and performed the MALDI-TOF MS of intact cells selected for various levels of intrinsic heterogeneity - human embryonic kidney (HEK293), mouse embryonic fibroblasts, human and mouse embryonic stem cells and primary cells from ovarian follicles of BALB/c and C57BL/6 mouse strains. Then, by rigorous statistical evaluation of the method robustness with respect to both technical and biological variability, we demonstrated that specific fingerprints of mammalian cells can be easily obtained with defined intra- and inter-instrumental overall reproducibility. Moreover, MS followed by advanced cluster analysis, revealed tiny phenotypic shifts in morphologically indistinguishable human embryonic stem cells induced to differentiation or undergoing silent phenotypic shifts in culture. In summary, intact cell MS represent a powerful tool for identification, authentication and phenotypization of cells in various applications, ranging from biomedical research, cell therapy to bioindustry.