Day 1 :
Keynote Forum
Marek M. Kowalczuk
University of Wolverhampton, UK
Keynote: Mass Spectrometry aimed at sequence analysis of natural and synthetic biodegradable macromolecules
Time : 09:00-09:25
Biography:
Marek M Kowalczuk received his PhD degree in 1984 from the Faculty of Chemistry, Silesian University of Technology, and DSc degree in 1994 at the same University. He was a visiting Lecturer at the University of Massachusetts in Amherst, MA, USA in 1990 and Marie Curie EU fellow at the University of Bologna, Italy. Currently, he is Professor at the University of Wolverhampton, UK 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. His research interests are novel mass spectrometry techniques for analysis of polymers at the molecular level, mechanisms of anionic polymerization related to the synthesis of biodegradable polymers possessing desired architecture, forensic engineering of biodegradable and functional polymers.
Abstract:
Mass spectrometry becomes nowadays a comprehensive analytical technique for copolymer characterization. Copolymers can exist in variety of co-monomer arrangements along the macromolecular chain such as block, gradient, random or alternating copolymers. Moreover, in addition to molecular mass distribution (MMD) the copolymers exhibit co-monomer composition distribution (CCD) and both these factors influence their physical properties for a given copolymer chemical composition. The aims of this study were focused on ESI tandem mass spectrometry of natural aliphatic copolyesters (PHA) and novel sequence-controlled copolymers obtained by anionic ring opening copolymerization of β-substituted β-lactones. Detailed analysis of these copolymers, including molecular chain architecture as well as chemical structure of the end groups, were established based on the ESI-MS/MS collision induced dissociation spectra. The arrangement of co-monomer structural units along the copolyester chains was demonstrated by comparison of the ESI-MS/MS fragmentation spectra and the respective fragmentation pathways. Moreover, the sequence distribution analysis of co-monomeric units confirmed the desired structure of novel sequence-controlled macromolecules. Understanding of the formation mechanisms of sequence-controlled copolymers and a detailed description of the individual reactions taking place at the initiation, propagation and termination of the copolymer chain growth are essential from standpoint of the relations between copolymer structure, properties and function. Thus, the ESI-MS/MS proved to be a favorable technique for structural analysis of the natural and synthetic biodegradable macromolecules.
Keynote Forum
Roger P Webb
University of Surrey, UK
Keynote: Ambient pressure mass spectrometry of fingerprints
Time : 9:30 - 10:00 AM
Biography:
Roger P Webb completed his PhD from Salford University and performed Postdoctoral studies at 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:
Ambient pressure mass spectrometry of latent fingerprints provides a potential route to the secure, high throughput and non-invasive detection of, amongst other things, drugs of abuse. We have shown that it is possible to detect both the drugs of abuse as well as the excreted metabolites in the fingerprints using ambient mass spectrometry. A recent press release on this work reported in the Analyst received a large amount of media interest around the world. We report here on the preliminary study employing MALDI, SIMS, DESI and paper spray backed up by GC-MS of oral fluids. The study shows that ambient mass spectrometry can detect cocaine, benziylecgonine (BZE) and methylecgonine (EME) in the fingerprints of drug users. The results provide exciting opportunities for the use of fingerprints a new sampling medium for secure, non-invasive drug detection. The mass spectrometry techniques used offer a high level of selectivity and consume only a small area of a single fingerprint, allowing repeat and high throughput analyses of a single sample.
- Track 4: Applications of Mass Spectrometry
Chair
Marek M. Kowalczuk
University of Wolverhampton, UK
Co-Chair
Miral Dizdaroglu
National Institute of Standards and Technology, USA
Session Introduction
Marek M. Kowalczuk
University of Wolverhampton, UK
Title: Mass spectrometry for forensic engineering of advanced polymer materials
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.
Sherry A. Tanumihardjo
University of Wisconsin, USA
Title: Mass spectrometric methods to support vitamin A policies in Africa
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 ïmol 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.
Pierluigi Mauri
Institute for Biomedical Technologies, Italy
Title: Mass spectrometry-based approach for clinical proteomics
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.
Pedro J. Sanches Filho
Instituto Federal Sul-rio-grandense, Brazil
Title: Determination of polycyclic aromatic hydrocarbons in sediments of rain drainage channels in urban zone (Pelotas - RS, Brazil)
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.
Pawel Jaruga
National Institute of Standards and Technology, USA
Title: Measurement of oxidatively modified DNA bases and nucleosides by isotope-dilution mass spectrometry
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.
Mehran F. Moghaddam
Celgene Corporation, USA
Title: Utility of Mass Spectrometry in Drug metabolism and pharmacokinetics (DMPK): Discovery of Small Molecule Drugs
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.
Joydip Das
University of Houston, USA
Title: Identification of alcohol binding residues in brain proteins using electrospray ionization mass spectrometry
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 PKCï¥ 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 PKCï¥C1A, His-248 and Tyr-250 of PKCï¥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 PKCï¥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 PKCï¥ 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.
Miral Dizdaroglu
National Institute of Standards and Technology, USA
Title: Measurement of DNA repair proteins in human tissues by liquid chromatography-tandem mass spectrometry with isotope-dilution
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.
Shawn R. Campagna
University of Tennessee, USA
Title: Application of metabolomics and kinetic flux profiling to understand the metabolic function of aquatic microbial consortia
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.
Fernando Ramos
University of Coimbra, Portugal
Title: Mass Spectrometry in Veterinary Drug Residue Analysis
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.
Anthony N. Gachanja
Jomo Kenyatta University of Agriculture and Technology, Kenya
Title: GC-MS-Application and Instrumentation Challenges in Eastern Africa
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.
Tânia Petta
University of São Paulo, Brazil
Title: Target and untargeted identification of oxylipids in biological samples by tandem mass spectrometry
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.
Vladimir Zaichick
Medical Radiological Research Centre, Russia
Title: Role and Place of Mass Spectrometry in Medical Elementology - a New Scientific Discipline
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.
Xiao Ding
Department of Drug Metabolism and Pharmacokinetics in Genentech, USA
Title: Quantitation of 14C-GDC-0032 (14C-taselisib) in human plasma by UPLC-accelerator mass spectrometry for the determination of absolute bioavailability of GDC-0032 in human
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%).
Scott M. Grayson
Tulane University, USA
Title: The use of ion-mobility spectrometry-mass spectrometry to elucidate polymer architecture
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.
- Track 1: Fundamentals of Mass Spectrometry
Chair
Roger Webb
University of Surrey, UK
Co-Chair
Christiane Auray-Blais
Université de Sherbrooke, Canada
Session Introduction
Roger Webb
University of Surrey, UK
Title: Ambient Pressure Molecular Concentration Mapping at Surfaces Using MeV-Secondary Ion Mass Spectrometry (MeV SIMS)
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.
Helmut Schwarz
Technische Universität Berlin, Germany
Title: Ménage-à -Trois: Single-atom Catalysis, Mass Spectrometry, and Computational Chemistry
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 ïƒ 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.
Athula B. Attygalle
Stevens Institute of Technology, USA
Title: Atmospheric Pressure Helium Plasma Ionization Mass Spectrometry
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.
Chrys Wesdemiotis
University of Akron, USA
Title: Multidimensional Mass Spectrometry Methods for Synthetic Polymer Analysis
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.
Rafael Garrett
Federal University of Rio de Janeiro, Brazil
Title: Ambient ionization MS for new insights in coffee science
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.
Christiane Auray-Blais
Université de Sherbrooke, Canada
Title: Mass Spectrometry Approaches for Translational Research: From Biomarker Discovery to Clinical Applications
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.