Day :
- Track 1: Applications of Mass Spectrometry Track 3: New Approaches in Mass Spectrometry Track 5: Recent Advances and Development in Mass Spectrometry
Chair
Claudia S. Maier
Oregon State University, USA
Co-Chair
Zhenhua GONG
Shanghai Jiaotong University, China
Session Introduction
Iulia M. Lazar
Virginia Tech, Biological Sciences, USA
Title: How to make MS data work best for biological research
Biography:
Iulia M. Lazar earned her Ph.D. in Chemistry from Brigham Young University in 1997. Following two postdoctoral appointments at Sensar Larson-Davis and Oak Ridge National laboratory, and a Principal Research Scientist position at The Barnett Institute/Northeastern University, she joined Virginia Tech in 2003. Presently, she is an Associate Professor with research interests focused on oncoproteomics, breast cancer cell cycle, signaling, biomarker discovery and the development of microfluidic and mass spectrometry technologies for the interrogation of biological systems. The findings of her research led to over 55 publications, book chapters, patents and numerous presentations at national and international symposia.
Abstract:
Mass spectrometry and proteomics have witnessed tremendous progress in the past two decades. From instrumentation and novel techniques and methodologies, to advanced software packages that handle large data sets, some of the most complex biological problems can be addressed today by capitalizing on the power of this technology. In our laboratory we develop proteomics approaches for the analysis of breast cancer cells. Our goal is to use a systems-level approach to uncover biological mechanisms that lead to uncontrolled cell proliferation, novel markers indicative of disease and protein clusters that can be used as effective therapeutic targets. In addition, we develop microfluidic instrumentation as a mass spectrometry front-end for high-throughput sample processing. In this presentation we will discuss the challenges that researchers face when dealing with the complexity of biological systems and the unique benefits offered by mass spectrometry in complementing genomics research. In particular, we will focus on the implications of achieving proteome depth, the complex dimension of posttranslational modifications, and proper timing of experimental conditions with biological relevance. Ultimately, we will discuss the potential of technologies such as mass spectrometry to transform our thinking about addressing problems that lie at the root of human disease.
Biography:
Dr. Peer earned his PhD in Pharmacology and Pharmaceutical Sciences in 2009, and more recently, a Masters in Pharmacometrics (2015). In 2009, he joined the Clinical Pharmacology Program at the National Cancer Institute as a postdoctoral fellow. Since 2012, Dr. Peer has been a Research Fellow and has overseen the bioanalytical operations and pharmacometric data analysis within the NCI Clinical Pharmacology Program. Dr. Peer has authored more than a dozen novel bioanalytical methods and contributed pharmacometric analyses to 24 additional publications. Dr. Peer’s research interests include bioanalytical chemistry, drug metabolism and transport, PK/PD modeling and simulation, pharmacogenomics, and pharmacometric analyses.
Abstract:
A sensitive and selective ultra-high performance liquid chromatography-tandem mass spectrometric method was developed for the quantification of temozolomide (TMZ) in nonhuman primate (NHP) plasma, cerebrospinal fluid (CSF), and brain extracellular fluid (ECF) following microdialysis. Ethyl acetate was used to extract the plasma and CSF samples, using theophylline as the internal standard (IS). ECF samples were diluted with acetonitrile prior to analysis. TMZ was separated on a Waters UPLC® BEH C18 column with an isocratic mobile phase of ammonium acetate (10 mM)-0.1% formic acid/acetonitrile (30:70, v/v) in a positive-ion multiple reaction monitoring mode (m/z 195.5→137.6 for TMZ; m/z 181.5→124.2 for IS). The retention time of TMZ and theophylline was 0.45 min with a total run time of 2.5 min. The method was validated over the range from 5–2000 ng/mL in NHP plasma, CSF, and ECF with respect to linearity, accuracy, precision, selectivity, and stability. This method was successfully applied toward the measurement of pharmacokinetic samples following various routes of drug administration.
Zhenhua GONG
Shanghai Jiaotong University, China
Title: Quantification of bilirubin on dry blood spot by tandem mass spectrometry
Biography:
Zhenhua GONG has completed his PhD at the age of 40 years from FUDAN University ,Shanghai China. He is pediatric surgeon, associated professor and director of a research team focusing on metabolic diseases and biliary atresia. He has published more than 10 papers in reputed journals and serving as an editorial board member of repute.
Abstract:
BACKGROUND: Hyperbilirubinemia is caused by many kind diseases and is harmful to neonate development, it is necessary to detect hyperbilirubinemia as early as possible, or probably detect biliary atresia (BA) earlier. A new method for screening and clinical determination of blood bilirubin on a dry blood spot (DBS) using tandem mass spectrometry (MS/MS) was developed. METHODS: The serum bilirubin controls and blood were prepared as DBS, extracted into a methanol solution containing isotope-labeled external standards, were subjected to HPLC, detect by MS/MS. Multiple-reaction monitoring of m/z 585—299 for unconjugated bilirubin(UBIL), m/z761—585 for monoglucuronide bilirubin(BMG) and m/z 937—585 for diglucuronide bilirubin (BDG) were used to detect bilirubins. RESULTS: The recoveries of UBIL by MS/MS were 90%-120.31% with an R2 value of 0.928 after linear regression (p<0.001). The coefficients of variation were less than 20%.The blood UBIL in neonates aged 3-7 days (68.14±21.56umol/L) was higher than in children aged older than 1 mouth(38.24±7.95umol/L). BMG and BDG were not higher in BA than control significantly. CONCLUSION: Quantification of UBIL on a dry blood spot by MS/MS is accurate, reliable and feasible for screening and clinical tests. There may be δ-bilirubin caused direct bilirubin elevated in BA.
Eduardo Gil-Santos
Université Paris Diderot, France
Title: High frequency nano-optomechanical disk resonators in liquids
Biography:
Eduardo Gil-Santos has completed his PhD at the age of 28 years old at the University Autonoma of Madrid, working in the BioNanoMechanics lab. He is currently a postdoctoral researcher at the University Paris Diderot, working in the OptoMechanics team of Matériaux et Phénomènes Quantiques laboratory. During his research career, he has focused on the study and development of micro- and nano-mechanical resonators, wih applications in chemical and biological sensing, biomedical and fundamental quantum physics. He has published more than 15 papers and has given more than 10 talks at international conferences.
Abstract:
Vibrating nano and micromechanical structures have been the subject of extensive research for the development of ultrasensitive mass sensors for spectrometry, chemical sensing and biomedical analysis. In short, the minimum detectable mass is proportional to the effective mass of the resonator and sensitivity improves if mechanical dissipation is reduced. Device miniaturization and dissipation control are therefore crucial. In liquids, the energy losses are high and the mass sensitivity is generally diminished dramatically. To circumvent this problem, novel structures have been proposed, such as micro-channels or micro-capillars where the liquid flows directly inside the resonators. While these structures indeed show lower mechanical dissipation, they can hardly be miniaturized. Here we demonstrate the potential of nano-optomechanical disk resonators in this context, in particular focusing on high-frequency radial breathing modes of these structures. Miniature semiconductor mechanical disks, with their high mechanical Q even in air (>103), their low mass (pg) and high mechanical frequency (GHz), present clear assets for mass sensing applications. However, they have not been operated in liquids so far. Here, we experimentally, numerically and analytically investigate the interaction of such vibrating disk resonators with arbitrary liquids, and propose models for both the frequency shift and dissipation of their mechanical modes. Nano-optomechanical disk resonators finally emerge as probes of rheological information of unprecedented sensitivity and speed, opening applications in high frequency sensing and fundamental science.
Katarzyna Pawlak
Warsaw University of Technology, Poland
Title: Coupling of micro-Electrochemical Reaction Cell to Electrospray MS (EC-ESI MS) for investigation of redox reaction products of cytotoxic metallocomplexes
Biography:
Katarzyna Pawlak has completed her PhD in the field of analytical chemistry at the age of 27 years from Warsaw University of Technology (WUT) and postdoctoral studies from Centre National de la Recherche Scientifique (CNRS, France) and Gdańsk University of Technology. She is the director of School of Liquid Chromatography and Hyphenated Techniques at WUT, vice-President of Polish Society of Mass Spectrometry and expert of National Cuncil of Supplenets and Nutritional Products. She has published more than 45 papers in reputed journals and has been serving as an editorial board member of repute.
Abstract:
Metallocomplexes are increasingly important in the search for more effective anticancer remedies. As hybrids of inorganic and organic components, metal complexes tend to assemble the advantages of inorganic and organic drugs, such as treating a broad range of tumors and a selective mode of action. A promising examples, indazolium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], auranofin (2,3,4,6-tetra-O-acetyl-1-thio-b-D-glucopyranosato-S-(triethyl-phosphine) gold(I)) or Au(III)bipyc ([(bipydmb-H)Au(OH)][PF6] (where bipydmb-H = deprotonated 6-(1,1-dimethylbenzyl)-2,2′-bipyridine)) are now appreciably progressing in clinical studies, with the outcome of few side effects and evidence of clinical activity. To explore the mechanism of the drug metabolism, its behavior in simulated physiological conditions should be examined. Studies carried out in vivo or even in vitro can be economically and time consuming, especially that oxidation or reduction can be induced by addition of specific reactive species which may interfere ESI MS signals. Another method – without interfering chemical composition - is application of electrochemical reaction chamber Antec's Roxy Reactor Cell (EC), which allows to obtain redox reaction at varied potentials. Structures of metallo-species obtained at the varied redox potential applied in EC have been studied by ESI MS. The metallodrug analysis was performed in a buffered solution of ammonium formate at the physiological pH of blood and at pH of gastric juice. Auranofin was found to be activated by loss of the glucose moieties, ruthenium complex by reduction of Ru(III) to Ru(II) and Au(III) complex by oxidation of pyridines. Obtained results are in agreement with present state of knowledge and make electrochemical chamber an interesting technique for studies of metallocomplexes metabolism.
Claudia S. Maier
Oregon State University, USA
Title: Tandem mass spectrometry for the characterization of peptide adducts of electrophlic lipoxidation products
Biography:
Claudia S. Maier, PhD, is currently a Professor in the Department of Chemistry at Oregon State University. She holds a PhD in Chemistry from the University of Konstanz, Germany. Her laboratory is concerned with the development and application of mass spectrometry for studying the response of biological systems to oxidative stress related to aging, chronic diseases and enviornmental exposures.
Abstract:
We report on the characterization and quantification of peptide adducts of reactive lipid peroxidation products using tandem mass spectrometric approaches using advanced quadrupole time of flight mass spectrometry platforms. The post-translational modification (PTM) of proteins by electrophilic oxylipids is emerging as an important mechanism that contributes to the complexity of proteomes. Protein carbonyls are recognized indicators of diverse pathological conditions associated to redox inbalance and oxidative stress. We will discuss chemoselectives probes in conjunction with advanced tandem mass spectrometry approaches for the detection, characteization and quantification of this class of analytes. We will demonstrate the use of multiple tandem mass spectrometry approaches including travelling wave ion mobility-enhanced tandem mass spectrometry for characterizing of isomeric peptide adducts of electrophilic oxylipids. Overall, we provide an update on mass spectrometric methods for the in-depth analysis of protein carbonyls, recognized markers of oxidative stress related post-translational modifications.
- Track 1: Applications of Mass Spectrometry Track 5: Recent Advances and Development in Mass Spectrometry Track 7: Fundamentals of Mass Spectrometry
Chair
Miral Dizdaroglu
National Institute of Standards and Technology, USA
Co-Chair
Jianjun Liu
Shenzhen Center for Disease Control and Prevention, China
Session Introduction
Yi Chen
Chinese Academy of Sciences, China
Title: Adjustable combinatory ionization “guns†for multiple tasks of analytical MS
Biography:
Yi Chen received his PhD in 1990 from Institute of Chemistry, Chinese Academy of Sciences (CAS), is a Professor of Analytical Chemistry, chairing Analytical Chemistry Department at University of CAS, and leading a research group on chemical perturbation analysis of cells and trace biological substances in living things. He has published 240 co-authored papers and 3 books and owned 20 patents. He is serving as Associated Editors and Editorial/Advisory Board Members of 15 journals, an associate member in Division V, IUPAC, and vice presidents of 5 Chinese academic societies including Professional Committee of Analytical MS in Chinese Chemical Society.
Abstract:
The resolution, accuracy and sensitivity of MS are above all governed by its ion source, which has activated a continuous innovation of new ion sources. However, there remain challenges to perform dynamic internal calibration, to couple MS with separation techniques, and to execute direct MS detection of complicated samples. To facilitate analytical MS, we have tried a combinatory use of miniaturized ionization techniques for a decade, with several “point-to” or gun-like ion-source devices designed and fabricated, e.g., nano-electrospray ionization, alternating/direct current discharge ionization, and laser head as well. These“guns”could be used individually or in combination, allowing performance of dynamic internal calibration for high accurate mass determination, with peak-height adjustable at any time during measuring. It is not need to stop the measurement in order to re-prepare a new set of peak-height-rational analytes or internal standards as usual. On a gold-coated photonic crystal surface, isotope-free internal standard ladders could be in situ superposed on an analyte for “Golden” internal calibrations. Mass-shift ionization was achieved in MALDI-TOF MS of small molecules in combined use of chemistry, which avoids the strong background in the low m/z range, but adopts the advantages of high mass resolution (several ppm) in the optimal range between 1000-2000 m/z by the aid of phthalocyanine derivatives. With some modifications, the“guns”could directly desorb and ionize some trace target analytes in complex soils and living plants and insects, or interface-freely cooperate with UPLC and CE.
Miral Dizdaroglu
National Iinstitute of Standards and Technology, Gaithersburg, USA
Title: Application of mass spectrometry in the discovery of small molecule inhibitors of DNA repair proteins as potential anticancer drugs
Biography:
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. 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:
Most therapeutic agents kill tumor cells by damaging DNA. Evidence shows that cancer cells increase their capacity to repair DNA by overexpressing DNA repair proteins. Increased DNA repair that may remove DNA lesions before they become toxic to cancer cells is a major mechanism for the development of resistance to therapy. DNA repair proteins constitute targets for inhibitors to overcome the therapy resistance. Inhibition of DNA repair proteins may help selectively kill cancer cells in combination therapy or monotherapy. Among DNA repair proteins, DNA glycosylases remove modified DNA bases in the first step of the base excision repair mechanism, paving the way for other proteins to fully repair DNA. We planned experiments to discover small molecule inhibitors of human DNA glycosylases NEIL1, OGG1 and NTH1. First, a fluorescence-based assay was developed to detect both glycosylase and lyase activities of these proteins. From a primary screen of ∼400,000 compunds, a number of inhibitors were identified. We applied GC-MS/MS with isotope dilution to determine the inhibition of NEIL1, OGG1 and NTH1 by identifying and quantifying the excised levels of their substrates. Four purine analogs were found to be potent inhibitors of the excision of NEIL1 substrates. Three of the four NEIL1 inhibitors also inhibited the exicision of NTH1 substrates, but not those of OGG1. Five other inhibitors were found to be potent inhibitors of the excision of OGG1 substrates; however, they displayed no inhibition of NEIL1 and NTH1 activities. Overall, this work forms the foundation for future drug discovery for DNA glycosylases.
Hui Xu
Guangzhou University of Chinese Medicine, China
Title: Determination of mycotoxin contamination in Traditional Chinese Medicines by LC-MS-MS
Biography:
Hui Xu has completed her PhD at the age of 34 years from Eberhard Karls University of Tuebeingen, Germany. She is a senior research fellow of Guangzhou University of Chinese Medicine and the vice director of Research Center of Chinese Herbal Resource Science and Engineering. She has published more than 10 papers in reputed journals.
Abstract:
Mycotoxins are toxic secondary metabolites from molds. Presently, liquid chromatography-tandem mass spectrometry (LC-MS-MS) has been widely used to evaluate mycotoxin contaminations in food and feed. However, application of LC-MS-MS to Traditional Chinese Medicines (TCMs) is difficult，because their complex matrices can cause matrix effect. Routinely, some purification procedures like immunoaffinity columns (IAC) should be included before injection to improve the accuracy and protect mass spectrometer. However, the high costs of columns restrict their application for large-scale screening. Therefore, the objective of our study is to develop a simple, rapid and cost-effective LC-MS-MS method for simultaneous determination of aflatoxins (AFB1, B2, G1 and G2) and sterigmatocystin (ST) in TCMs. The method is based on single extraction with (84/16, v/v) acetonitrile-water then analysis of the diluted crude extract without further clean-up. The chromatographic separation was achieved on a C18 column, with a mobile phase gradient prepared from aqueous 4 mmol L-1 ammonium acetate-0.1% formic acid and methanol. Quantification of the analytes was by selective reaction monitoring (SRM) on a triple quadrupole mass spectrometer in positive ionization mode. Special focus was on investigating and reducing matrix effects to improve accuracy. The established method was validated by determination of linearity, sensitivity, extraction recovery and precision. Comparison with IAC pretreatment demonstrated both methods are comparable in respect of the recovery of aflatoxins. Finally, the validated method was used to evaluate mycotoxin contamination in total 294 samples of 30 TCMs collected from local hospitals and pharmacies and the results obtained were analyzed.
Jianjun Liu
Shenzhen Center for Disease Control and Prevention, China
Title: Proteomic study of SET-mediated abnormal protein phosphorylation and histon modification in trichloroethylene-induced hepatic cytotoxicity
Biography:
Jianjun Liu has been studying the mechanisms of TCE-induced hepatotoxicity for several years. She is a doctoral supervisor. She is the director of Key Laboratory of Modern Toxicology of Shenzhen, a division of Shenzhen Center for Disease Control and Prevention. She has published more than 60 papers in international and chinese journals and has authorized 3 patents of invention.
Abstract:
Trichloroethylene (TCE) was widely used in industrial productions and turned into an environmental and occupational toxicant. Expression of SET protein was previously found as dose-dependent with TCE in human liver cells. It is also an important inhibitor of phosphatase 2A and histone acetyltransferase. However SET-mediated abnormal histone modification and protein phosphorylation in TCE induced hepatic cytotoxicity remain poorly understood. SET-mediated protein phosphorylation in TCE-induced hepatic cytotoxicity was analyzed by iTRAQ labeling and IMAC enrichment based quantification proteomics study. 14 phosphopeptides from 13 proteins were found as SET-mediated (de)-phosphorylation in hepatic cytotoxicity of TCE. Furthermore, nucleolin was found self-regulating by SET-mediated phosphorylation through enhanced interaction with c-myc and inhibiting of c-myc. SET-mediated histone methylation and acetylation were analyzed by TAU-SDS-PAGE seperation combining with LC-ESI-MS based label-free quantification. The ubiquitinated and sumoylated histones were first enriched by specific antibodies then labeled with stable isotopic di-methylation reagents and analyzed by LC- ESI-MS. 12 acetylated peptides, 11 methylated peptides, 10 ubiquinated peptides and 6 sumoylation peptides were found as SET-mediated alteration in hepatic cytotoxicity of TCE. Our findings provided molecular-level evidence further supporting our previous findings that knockdown of SET attenuated TCE-induced hepatic cytotoxicity. SET-mediated self-regulating of nucleolin and abmornal histone modifications further revealed the molecular mechanism of SET-mediated hepatic cytotoxicity of TCE.
Youssef Mouneimne
American University of Beirut, Lebanon
Title: Assessment of BPA levels in urine by HPLC MS
Biography:
Dr. Youssef Mouneimne has completed his Dr of engineering (1984) and Ph.D in applied chemistry (1986) from Claude Bernard Lyon I University. He worked as research scientist at the “centre de biophysique moleculaire, CNRS” in Orleans , then at Texas A&M university, Baylor and Harvard University. He invented the electroinsertion of proteins into cell membrane and the flow electroporation system for drug delivery, among many other inventions. Actually he is the director of the Central research laboratory at the American university of Beirut. He is actually involved in many research project including extraction of biofuel and other chemicals from algae, analysis of BPA and sodium in food, Benz[O]Pyrine metabolomics.
Abstract:
Bisphenol A (BPA), is a synthetic chemical used in the production of plastics and epoxy resins of food packaging. BPA may be associated with adverse health effects. The objective of this study was to assess the level of BPA among a representative sample of Lebanese population residing in Beirut, as well as to assess factors associated with these levels. A representative sample of 501 men and women aged 18-79 years was examined in a cross-sectional manner with a health and food questionnaire, anthropometric measures and urine BPA analysis. Spot urine samples were collected in glass jars, aliquoted in glass containers and frozen at -20oC for further analysis. High performance liquid chromatography-mass spectroscopy was used to analyze BPA according to Coughlin et al. (2011) and Ning et al. (2011). We found a mean urine BPA concentration of 3.67 ppb, with a standard deviation of 4.75. Some samples measured high concentrations up to 59.7 ppb, giving the total pattern a skewed distribution. We conclude that the concentration found are comparable to the values found in other countries.
Biography:
Gongke Li obtained her PhD degree in Analytical Chemistry from Sun Yat-sen University of China in 1992. She then joined Sun Yat-sen University and became a Professor since 2000. With expertise in chromatographic analysis and spectral analysis, her major research interests are focused on sample preparation techniques, analytical techniques for trace analysis of complex samples and coupling device for online analysis. She also studies both analytical- and preparative-scale separations methods for natural products. She has published more than 300 research papers in reputed journals, 13 authorized invention patents and 1 monograph. She was awarded the Chinese women's analytical chemist in 2015. She is currently the Associate Editor of Journal Separation Science.
Abstract:
Cytokinins play a critical role in controlling plant growth and development, but it is difficult to be determined in plant samples due to the extremely low concentration level of pmol/g. So it is important of efficient sample preparation with selective enrichment and rapid separation for accurate analysis of cytokinins. Herein, a supramolecular perhydroxy-cucurbit[8]uril (PCB[8]) was fabricated into the Fe3O4 magnetic particles via chemical bonding assembly and magnetic perhydroxy-cucurbit[8]uril (MPC) materials were obtained. The MPC had good enrichment capability to cytokinins and the enrichment factors were more than 208. The interaction of MPC and cytokinins was investigated by adsorption test and density functional theory (DFT) calculation. The MPC was used as sorbent of magnetic solid-phase extraction for the analysis of cytokinins in plant samples. A sensitive and selective UPLC-MS/MS method was developed with low detection limits of 0.14-0.32 ng/L for cytokinins analysis. Five cytokinins including zeatin riboside, meta-topolin, kinetin, kinetin riboside and zip with 6.12-87.3 ng/kg were determined in the soybean sprout and arabidopsis thaliana. The recoveries were in the range of 76.2-110% with RSDS of 2.3-9.7%. The magnetic perhydroxy-cucurbit[8]uril materials with selective enrichment capability have good potential for analysis of ultratrace targets from complicate sample matrix.
- Track 1: Applications of Mass Spectrometry Track 5: Recent Advances and Development in Mass Spectrometry Track 8: Ionization Techniques
Chair
Sherry A. Tanumihardjo
University of Wisconsin, USA
Co-Chair
Hui-FenWu
National Sun Yat-Sen University, Taiwan
Session Introduction
Christiane Kruse Fæste
Norwegian Veterinary Institute, Oslo, Norway
Title: Development of liquid chromatography-tandem mass spectrometry methods for the quantitation of Anisakis simplex proteins in fish
Biography:
Christiane Kruse Fæste has completed her PhD in biochemistry at the age of 28 from the Leibniz University in Hanover, Germany, and the Max-Planck Institute for Plant Breeding Research in Cologne, Germany. She has been working in Preclinical Pharmaceutical Industry and is now expert for xenobiotics metabolism, toxicokinetics, food allergy and proteomics in the Section of Chemistry and Toxicology of the Norwegian Veterinary Institute, Oslo, Norway. She is member of the Norwegian Committee for Food Safety (panel on contaminants), CEN (Food Allergens), AOAC (Food Allergens), EU-Research Executive Agency (Chemistry), ILSI (Allergens) and COST (Allergens). She has published 40 articles on food allergens and mycotoxin metabolism.
Abstract:
Larvae of the parasite Anisakis simplex are present in many marine fish species used for food. They infect mostly the gut and inner organs but have also been shown to penetrate into the fish fillet. Thus, human health can be at risk, either by contracting anisakiasis through the consumption of live parasites in raw or under-cooked fish, or by sensitisation to anisakid proteins in processed food. Methods for the detection of anisakid larvae include visual techniques and PCR, and immunological assays for protein determination. Recently, mass spectrometry-based proteomics has been used for the characterisation of A. simplex proteins, preparing for the development of two quantitative liquid chromatography-tandem mass spectrometry methods in the present study. Both, the label-free semi-quantitative nLC-nESI-Orbitrap-MS/MS (MS1) and the heavy peptide-applying absolute-quantitative (AQUA) LC-TripleQ-MS/MS (MS2) use unique reporter peptides derived from anisakid hemoglobin and SXP/RAL-2 protein as analytes. Standard curves in buffer and in salmon matrix showed good linearity and sufficient sensitivity for the intended method use. Preliminary validation included the assessment of specificity, repeatability, reproducibility, and applicability to incurred and naturally-contaminated samples for both assays. By further optimisation and full validation the LC–MS/MS methods could be standardised and used as confirmative techniques for the detection of A. simplex protein in fish and products.
Bernard Do
University of Paris-Sud, France
Title: Photo-reactivity assessment of perfusion drugs combining quantum chemical approach and experimental studies
Biography:
Dr Bernard Do has completed his PhD at the age of 29 years from Paris-Descartes University and postdoctoral studies from Paris-Saclay University School of Pharmacy. He is hospital pharmacist at Assistance Publique-Hôpitaux de Paris, associate professor at Paris-Descartes University and senior researcher at Paris-Saclay University focusing on drug intrinsic stability and drug/polymer interactions. He has published more than 45 papers in reputed journals and has been serving as an editorial board member of repute.
Abstract:
The susceptibility of a drug substance or drug product to degrade upon light exposure is far from being an uncommon property. Actually, the number of drugs found to be photo-chemically unstable is steadily increasing and the European Pharmacopoeia recommends light protection for hundreds of medical drugs and a number of adjuvants. The evaluation of interactions between drugs and light accounts for a natural part of the research and development work for new medical products. This allows the formulations containing drugs susceptible to photoreactions to be marked and adequately stored. However, in some situations, the ideals are not always maintained. Indeed, it is well known that in-use conditions do not always coincide with the control or design spaces, within which, it has been shown that the drug is stable. In this context, liquid preparations are much more concerned than solid formulations of the corresponding drug substances, being usually far more photolabile. That's why elucidating photodegradation mechanisms of drug products can be of paramount importance in that this can help reckon whether special procedures or additives could be used to prevent any loss of drug potency and formation of photoproducts during their handling and administration. We present here an approach combining a computational method based upon the density functional theory (DFT) and experimental studies using liquid chromatography-multistage mass spectrometry (LC-MSn) to investigate the photodegradation behaviour of parenteral drugs through two case studies, tirofiban and raltitrexed. The results predicted by DFT were experimentally supported through the photoproducts identification. As their photoreactions were shown to mainly proceed through type I and type II photosensitization mechanisms, the possibility of adding quenchers to the formulation can be considered. The major contenders would be substances such as ascorbic acid, a-tocopherol, and BHT, which are capable of acting as free radical scavengers and weak singlet oxygen quenchers.
Hui-Fen Wu
National Sun Yat-Sen University, Taiwan
Title: Nanomaterial based mass spectrometry applied on pathogens, cancer study and single cell detection
Biography:
Hui-Fen Wu currently is a distinguished Professor in National Sun-Yat-Sen University. She obtained PhD from University of Texas at Austin, USA (1994). She was a faculty in Tamkang University for 10 years. In 2006, she joined National Sun Yat-Sen University. Her research work is focused on applications of various nanomaterials for pathogenic bacteria analysis and cancer study. She has been served as Editorial Board Members for 15 journals. To date, she has published 205 scientific journals, 250 conference presentation, 13 book chapters and 7 patents.
Abstract:
We integrated nanotechnology, microbiology and mass spectrometric techniques for biomedicine studies including pathogenic bacteria analysis, cancer study, single cell detection and also developing biochips. Our various biomedicine projects have been successfully conducted and also briefly described as below: rapid detection of in vivo kinetics of pathogenic bacterial infection in mouse blood and urine using MALDI-MS; Nanoparticle (NP) assisted MALDI-MS as bacterial biosensors for rapid analysis of yogurt; ZnO and Ag nanoparticles as bifunctional nano probes for bacterial detection, CdS QDs for degradation of extracellular polymeric substance (EPS) of E.coli; mass spectrometry as a bacterial biosensor using TiO2 NPs assisted MALDI-MS; tracing the Staphylococcus aureus on ants using physical preconcentration coupled ZnO NP assisted MALDI-TOF MS. The bacterio-toxicity/compatibility of Platinum nanospheres, nanocuboids and nanoflowers with different size effect was demonstrated. We also have developed bifunctional titania (Ti) chip for highly sensitive pathogenic bacteria analysis in the MALDI-MS. Recently, we also applied gold nano-platform mediated microwave digestion in detecting single cancer cells and cancer stem cells analysis using MALDI-MS. We also successfully applied platinum nanoparticles for photothermal treatment of neuro 2A cancer cells. All these studies show the successful implementation of nanotechnology into mass spectrometry for pathogen analysis and cancer study. Our approaches can be applied as rapid, direct, effective and sensitive techniques for future microbiology and cancer analysis especially in clinical and medical studies.
Guo-An Luo
Macau University of Science and Technology, China
Title: Study on free fatty acids metabolome using 2D nano LC-NSI-Q-TOF/MS, application to polycystic ovarian syndrome in women
Biography:
Guo-An Luo obtained the Master degree from East China University of Science and Technology in China in 1982. He is the professor of Tsinghua University and Macau University of Science and Technology. He has long been engaged in pharmaceutical analysis, the systems biology for traditional Chinese medicine (TCM) and modernization for TCM research. Six research monographs and more than 760 papers were published, of which 310 was indexed by Web of Science, with an H-index of 35. Prof. Luo has been authorized more than twenty-five invention patents and completed six preclinical applications of new drug.
Abstract:
Free fatty acids (FFAs) including omega-3 and omega-6 polyunsaturated fatty acids were major derived from arachidonic acid (AA) and eicosapentaenoic acid (EPA), which plays important roles in many physiological processes, especially inflammation. Assessment of eicosanoids is important for understanding their homeostatic and pathophysiological roles in inflammatory disease. A new analytical approach focus on polyunsaturated fatty acids metabolome was established using isotope dilution, solid phase extraction and 2D nano liquid chromatography-nanospray ionization-time of flight-mass spectrometry (2D nano LC-NSI-Q-TOF/MS). Within 35 min, 58 FFAs, including free fatty acids, prostaglandins, lipoxins, resolvins, leukotrienes, thromboxans, maresins, hydroxyeicosatetraenoic acids as well as epoxyeicosatrienoic acids were quantified along with seven corresponding isotope dilution internal standards. The limits of quantification were between 0.008 and 20 pg per injection for 1ïL. The efficiency of separation were perfect, all of the standards were separated well (Figure 1). The method validations for linearity, accuracy, precision, recovery were satisfied. Besides, this approach was performed to analyze both of targeted and non-targeted metabolomics studies in serum samples of women with polycystic ovarian syndrome (PCOS) and health women, the results indicated that the significant differences noted in the levels of heptadecanoic acid, mysristic acid, pentadecanoic acid, 9E, 11E-octadecadienoic acid, arachidolic acid, palmitic acid, oleic acid and stearic acid (p < 0.001) as well as palmitelaidic acid (p < 0.01) (Figure 1). Women with PCOS may have a unique metabolomic finger print and a definitive study is feasible. These findings may indicate that the PCOS were positive correlation between patient and health woman in fatty acids syntheses.
Sherry A. Tanumihardjo
University of Wisconsin, USA
Title: Natural Abundance of 13C in Serum Retinol Differentiates between Dietary Intake of C3 versus C4 Plants
Biography:
Tanumihardjo studies vitamin A and carotenoid metabolism, serves as director of the Undergraduate Certificate in Global Health, and teaches at undergraduate and graduate levels including international field experiences. She is on the Executive Board for the UW Global Health Institute. Tanumihardjo has more than 150 publications and chapters. She has presented at more than 250 domestic and international venues. 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, Dannon Creative Leadership Institute, Endowed Chair and Vilas Associate at UW.
Abstract:
Vitamin A is a micronutrient essential in vision, reproduction, immune function, and cellular differentiation. Provitamin A carotenoids are plant sources of vitamin A. The isotopic distribution of 13C and 12C in humans is determined by what foods are consumed. C3 plants, i.e., green vegetables, carrots, and pumpkins, have lower 13C:12C than C4 staple crops, i.e., maize, sorghum, and millet. Vitamin A foods from corn-fed animals will reflect the 13C:12C feed that the animals eat. The serum retinol 13C:12C was previously evaluated as a biomarker for vegetable intake. The retinol 13C:12C decreased in humans who increased their vegetable intake (range -26.21 to -31.57‰, P = 0.050) and correlated with provitamin A carotenoid intake (P = 0.079). The average ï¤ï€ difference was -0.526 with increased vegetable intake, while control increased by +0.370. A 2X2X2 study in Mongolian gerbils fed white and orange maize or carrots for an extended period of time. Serum retinol δ13C‰ differentiated between those consuming white maize and white carrots (–27.1±1.2 δ13C‰) from those consuming orange maize and white carrots (-21.6±1.4 δ13C‰, P<0.0001) and white maize and orange carrots (-30.6±0.7 δ13C‰, P<0.0001). This method was applied to Zambian children who had been fed either orange or white maize for two months. Those children who consumed orange maize had a lower δ13C‰ (-26.64±1.98) than their white maize-consuming counterparts (-27.39±1.94) (P = 0.049). In the application of this methodology to efficacy or effectiveness trials, it will be important to choose the appropriate control group and number of subjects for comparison analyses.
Kihyung Song
Korea National University of Education, Republic of Korea
Title: Gas phase fragmentation mechanisms of protonated testosterone as revealed by chemical dynamics simulations
Biography:
Kihyung Song has completed his PhD at the age of 39 years from Texas Tech University. Since 1989, he have been a Professor of Department of Chemistry at Korea National University of Education. He is the chairman of the department now. He has published more than 100 papers in reputed journals.
Abstract:
We have studied the fragmentation mechanisms leading to ions produced by collision-induced dissociation of protonated testosterone in the gas phase. At this aim we have used QM+MM chemical dynamics simulations with semi-empirical Hamiltonian for the description of testosterone ion fragmentation. Results show that MSINDO method is able to correctly produce the typical peaks obtained experimentally for protonated testosterone. Simulations also provide mechanisms. In particular we discussed those providing the typical testosterone peaks, m/z 97, 109 and 123 and compare with what suggested experimentally. Finally, we rationalized the appearance of different peaks in terms of their dynamical behavior. This study shows for the first time that chemical dynamics can be used to rationalize steroid gas phase fragmentation, thus paving the way for using this approach as complementary tool in doping detection.
Matthew S. Klee
VP Market Development Dani Instruments, Inc. USA
Title: Getting the Most out of Capillary Gas Chromatography
Biography:
Matthew S. Klee is internationally recognized for contributions to the theory and practice of gas chromatography. His experience in chemical, pharmaceutical and instrument companies spans over 30 years. During this time, Dr Klee’s work has focused on elucidation and practical demonstration of the many processes involved with GC analysis, with the ultimate goal of improving the ease of use of GC systems, ruggedness of methods and overall quality of results.
Abstract:
This course is designed for experienced GC users who seek an enjoyable, practical, and focused update on the latest developments in capillary gas chromatography. Attendees will receive an informed perspective on current “best practices” and the latest trends and advancements in capillary gas chromatography. Traditional GC theory will be expanded and applied to everyday situations. Perspective will be shared to differentiate the marketing hype from that which can make a real difference in the lab. Straightforward and practical explanations will be presented for each topic such that students will be able to make informed decisions on adoption and application of new techniques and instrumentation.
- Track 2:High-performance liquid chromatography (HPLC)
- Track 1: Applications of Mass Spectrometry Track 4: Advances in Sample Preparation Techniques Track 11: Advances in Chromatography
Chair
Sam F.Y. Li
National University of Singapore, Singapore
Co-Chair
Kalluru Seshaiah
Sri Venkateswara University, India
Session Introduction
Kalluru Seshaiah
Sri Venkateswara University, India
Title: A Rugged Bio-analytical Method for Determination of Hydralazine in Human Plasma using Gas Chromatography coupled with Tandem Mass spectrometry
Biography:
K. Seshaiah has completed his PhD from Sri Venkateswara University, Tirupati, India and postdoctoral studies from Chaoyang University of Technology, Taiwan (ROC). He is the Chairman, Board of Studies in Chemistry, Sri Venkateswara University, India. He has guided 20 Ph.D., students and published more 80 papers in reputed journals. He has completed several research projects sponsored by DST, CSIR, UGC, Govt. of India and has international collaboration with University of Surrey, UK.
Abstract:
A simple, sensitive and rugged quantitative method for the determination of Hydralazine in human serum by gas chromatography-tandem mass spectrometry (GC-MS/MS) has been developed and validated. Hydralazine (1-hydrazinylphthalazine) is a vasodilator used to treat severe hypertension, congestive heart failure, myocardial infarction and preeclampsia. The HPLC methods reported for assays of Hydralazine in plasma are complicated because of rapid cooling and or separation of plasma immediately after withdrawing each blood samples, and Donnell et al.,1 and Woodworth et al.,2 have reported about the instability of Hydralazine at physiological pH. In the present study, a simple, specific and selective method has been developed for the determination of Hydralazine from plasma samples using GC-MS-MS. The sample preparation technique, described here involves rapid processing of samples and also rugged according to commonly acceptable FDA guidelines. Chromatographic separation has been achieved by GC with MS detector in positive ionization mode. Helium was used as the carrier gas and DB 5ms column, (30m X 0.25mm, 0.25µm (J& W; DB 5ms) film thickness Capillary column coated with cross Linked Polymer was used) gave a good peak shape for both analyte and internal standard. The retention time of Hydralazine and 4-methyl Hydralazine (IS) were 12.99 and 14.17 minutes respectively. The total run time of 20.00 min for each sample was freezed upon. A simple liquid-liquid extraction (LLE) technique was employed for the sample preparation that provided a good recovery of both analyte and IS. The LLOQ was 10.000ng/mL Hydralazine hydrochloride. The precision and accuracy of analyte at LLOQ concentration was found to be 6.423 % and 88.093 %, respectively. The CV % was 3.21% and 3.81% and % accuracy was 97.82% and 100.373% for LQC and HQC respectively.
Sam F.Y. Li
National University of Singapore, Singapore
Title: NMR and MS-based Metabolomics of Microcystis spp. Unravel the Involvement of Microcystin in Thylakoid Membrane Stabilization and Photoinhibition Resistance
Biography:
Professor Sam Li is a faculty member at the Department of Chemistry, National University of Singapore. He received his BSc, PhD and DSc degrees from Imperial College, UK. His research interests include environmental science and technology, biosensors, metabolomics and nanotechnology. He has authored/co-authored 325 publications in international peer review journals, more than 100 conference presentations and 10 US patents. He serves/served on editorial advisory boards of several international scientific journals, including Electrophoresis and Journal of Chromatographic Science.
Abstract:
Microcystin (MC), a potent hepatotoxin commonly produced by several freshwater cyanobacterial species, especially Microcystis, pose serious threats to water safety worldwide, making them unsuitable for recreational uses, as well as drinking water supplies. Here, the metabolomes of MC-producing and MC-free Microcystis species chronically exposed to strong light illumination were compared using complementary metabolomics tools; NMR and LC-MS. NMR and MS-based metabolomics were capable of revealing MC-influenced metabolic changes in Microcystis spp. The up-regulation of thylakoid membrane building blocks (phosphotidylglycerol), photosynthetic pigments (chlorophyll-a), and carbon reserve sources (sucrose, glycogen, 3-hydroxybutyreate) in toxic Microcystis strains, indicated the involvement of MC in photosynthesis apparatus stabilization under high light illumination. The lowered levels of intermediates involved in phospholipid degradation revealed that Microcystis strains were more resistant to light-induced oxidative stress in the presence of MC. Taken together, our comparative metabolomic data suggest a photoinhibition resistance role for MC in light-stressed Microcystis spp., and confirm its involvement in thylakoid membrane and chlorophyll-a stabilization..
Lilian Regina Barros Mariutti
University of Campinas, Brazil
Title: Carotenoids and carotenoid esters from fruits: strategies for identification by LC-MS
Biography:
Lilian Mariutti is a Researcher in the Department of Food Science of the Faculty of Food Engineering at University of Campinas (Brazil). She received her Doctorate degree in Food Science from Unicamp in 2009. She also holds a M.S. in Food Science and B Sc in Food Engineering. She was a Researcher Fellow in the Laboratory of Veterinary Drug Residues of the Brazilian Ministry of Agriculture, Livestock and Food Supply. Her research focuses on the identification and bioaccessibility of bioactive compounds (carotenoids and phenolic compounds), antioxidant activity of bioactive compounds in different systems and action of natural antioxidants in food.
Abstract:
Carotenoids are yellow to red liposoluble pigments found in plants, animals and microorganisms. The health benefits of the carotenoids have been attributed mainly to their antioxidant properties, which would minimize the in vivo oxidative damages induced by reactive oxygen (ROS) and nitrogen species (RNS). Their basic structure is a tetraterpene with a series of conjugated double bounds, and they can be divided into two main groups: carotenes (composed only of carbon and hydrogen atoms) and xanthophylls (contain at least one oxygen atom). Xanthophylls possessing hydroxyl groups can be found in nature acylated to several fatty acids, generating the carotenoid esters. The fact that a single carotenoid can be found acylated in several different combinations with distinct fatty acids further increases the variability of structures that can be found and the complexity of the analyzed matrix. Due to the high number of possible carotenoid esters in fruits and since all the esters of the same free carotenoid present exactly the same UV-visible spectrum, their identification must include tandem mass spectrometry (MSn) studies. Therefore, separation of carotenoids in reversed-phase liquid chromatography using C30 columns in combination with DAD and APCI-MS/MS detection allows high sensitivity and selectivity and is the technique of choice for carotenoid identification in complex matrices. Sample preparation and strategies for the identification of carotenoids and carotenoid esters based on UV-vis, MS and MS/MS spectra will be presented and discussed.
Biography:
Lokesh Gupta, has completed his PhD at the age of 26 years by researching in University of Delhi and Ch. CS University Meerut, India. He is an analytical research scientist and serving as Chief Manager of Analytical R&D team with TEVA API (a world leader in generic pharmaceuticals) in India, focusing on analytical research/methods development, spectroscopic characterization of organic pharmaceuticals, technology validation, impurity isolation and characterization etc. He is participating and discussing his commended research in several national/international seminars/conferences. He had published more than 42 papers in peer reviewed reputed journals of chemistry & spectroscopy and serving as an eminent referee for several journal of international repute.
Abstract:
From decades spectrometry has been accepted as a key analytical tool for understanding and characterization of molecules in chemistry, the level of world-wide research activity in this field promises that these capabilities will continue to improve, rapidly. Over the period of time sensitivity of spectroscopy tools have been improved to work at micro level and in more depth, ultimately become a faster research tool; such tools now being used as process analytical tools (PAT) giving online understanding of characteristics of a molecule during research/development and in production. Same time their utilization has widen up by coupling these tools with liquid chromatography, and thermal measurement tools e.g. (LC-MS-MS, LC-NMR, DSC-FT etc.)Mass spectroscopy has achieved horizons from single quadrupole to triple quadrupole, MS-TOF & the Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) these have become a powerful and widespread analytical tool in life science and health sector. The dynamic mass range (1-300 kDa), high accuracy and sensitivity make it a superior method for analysis of all kinds of biomolecules including proteins, nucleic acids, metabolites and carbohydrates. Particularly in drug discovery, where compound identification and purity from synthesis and early pharmacokinetics are determined, MS has proved indispensable outcomes. Today, the MS practitioner can choose among a range of ionization techniques which have become robust and trustworthy on a variety of instruments with demonstrated capabilities. In combination with 2D-elelectrophoresis, MALDI-TOF-MS is particularly suitable for the identification of protein spots via mass fingerprint or micro sequencing. Same time MS-TOF is widely used in pharmaceutical word, TOF has improved the sensitivity by increasing the path length in TOF tube, so ion remain in path of light for longer time increasing sensitivity, on other side measurement tolls has been improved to see the mass number in several digits can differentiate molecules having closure mass and differentiating isotopes. Software calculates elemental formula, for which confirmation performed by comparing theoretical fragments to the obtained TOF -MS/MS of molecule. In this review I had evaluated and focused on advancement and updates in MS field, with respect to technology update & applications.
Hong Wang
Shanghai Institute for Food and Drug Control, Shanghai, China
Title: Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH Extracted from Human Urine
Biography:
Hong Wang received her PhD degree in 2012 from the Institutes of Biomedical Sciences, Fudan University, China. Using MS and a novel lectin microarray, she revealed the glycosylation of biomarkers. She currently studies the quality of biotechnology drugs in Shanghai Institute of Food and Drug Control. Based on the innovation of MS, she contributes to the analysis of protein glycosylation, the structural characterization of mAbs, the consistency of biosimilars. She is the leader of Shanghai Sailing Program. She has taken part in several national and provincial R&D programs. She has published more than 10 scientific papers in high level journals.
Abstract:
Human follicle-stimulating hormone (hFSH) plays a key role in the development and function of the reproductive system. Two classes of hFSH-containing pharmaceutical preparations currently exist; those derived from the urine of postmenopausal women (uhFSH) and those manufactured using recombinant DNA technology (rhFSH). Comparative clinical studies have revealed the differences in oocyte quality and clinical outcome between rhFSH and uhFSH. The oligosaccharide moiety is critical in determining the pharmacological properties of therapeutic glycoproteins. hFSH is a glycoprotein and glycan significantly influences the biological properties of hFSH. Considering the importance of hFSH glycosylation to the biological activity of hFSH, evaluating the difference in glycosylation between rhFSH and uhFSH intended for clinical use is essential. Using a glycoproteomic strategy, this study compared the glycosylation of uhFSH with that of rhFSH. Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman−Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. Results showed that highly sialylated, branched, and macro-heterogeneity glycans are predominant in the uhFSH compared with those in rhFSH. A high degree of heterogeneity was observed in the N-glycopeptides of both hFSHs. The O-glycopeptides of both hFSHs, which have not been described previously, were characterized herein. The differences in glycosylation provide useful information in elucidating and in further investigation the critical glycan structures of hFSH.
Mahmoud A. Saleh
Texas Southern University, USA
Title: Mass Spectrometry Regiospecific Analysis of neutral triacylglycerols
Biography:
Mahmoud A Saleh has completed his PhD in physical organic chemistry at the age of 28 years from University of California, Davis and postdoctoral studies at University of California, Berkeley in environmental toxicology. He is a professor of chemistry at Texas Southern University. He has published more than 150 papers in reputed journals and has been serving on editorial board member of several journals.
Abstract:
Different mass spectrometry ionization techniques were compared for their use in regiospecific analysis of neutral triacyglycerol (TAG) in different samples of seeds oil. Atmospheric pressure chemical ionization, atmospheric pressure photo ionization, electro spray ionization, ion trap and QTOF accurate mass were used in the LCMS experiments. Electron ionization, chemical ionization and QTOF accurate mass were used in GCMS experiments. MSMS QTOF and MSMS1-n ion trap experiments were used for structural conformation. Seed oil of Silybum marianum and 9 species of the Apiaceae family (dill, caraway, cumin, coriander, anise, carrot, celery, fennel and Khella) were used in this investigation. Results for regiospecific analysis showed a non-random fatty acids distribution in both S. marianum and Apiaceae seeds. Petroselinic acid was predominantly located at the sn-1 and sn-3 positions in carrot, celery and parsley seed oils, while it was mainly at the sn-2 position in caraway seed oil. The distribution of linoleic acid was opposite to that of petroselinic acid. Oleic acid was mostly located at the sn-2 position, except for caraway, where it was evenly distributed between the sn-1(3) and sn-2 positions. Both the saturated FA, palmitic and stearic acid, were located mainly at the sn-1(3) position. Based on our finding, it can be concluded that fragmentation pattern for TAGs follow the following scheme: The most abundant ions observed in the mass spectra of TAG are positive ions containing two complete fatty acid chains and those containing only one complete chain corresponding to the loss of one or two fatty acids respectively. Two ions were found corresponding to the ion remaining after the loss of one acyloxy group from the molecular ion [M-RCOO]+, and the other corresponding [M-RCOOH]+. If the three fatty acids are different, three different [M-RCOO]+ and [M-RCOOH]+ ions corresponding to the loss of each different acyloxy group. However, due to steric and electronic property, the loss of the fatty acid located in the sn2 is energetically less favorable than one at the end positions (sn1 and sn3). Therefore, ions corresponding from the loss of fatty acids from the sn1 and sn3 positions are much more abundance the ion corresponding to the loss of fatty acid at the sn2 position.
Oana Teodora Marisca
Babes Bolyai University, Romania
Title: An alternative diabetic retinopathy therapy: a laser triggered microsystem for controlled release of Resveratrol
Biography:
Oana Teodora Marisca is a 25 years old PhD student at Babes Bolyai University, Romania. Her research focuses on gold nanoparticles-cell interaction and targeted therapies using gold and silver nanoparticles. She is currently member of a Romanian government funded research project entitled “A controlled-release target delivery system for the alternative therapy of retina diseases-a novel perspective”. She published 4 articles in reputed journal
Abstract:
A severe ocular complication that causes visual impairment and even worse, blinding, due to a damage of the retina and the optic nerves is diabetic retinopathy (DR). During recent years, there was shown a significantly increase in using natural Resveratrol (RV) (3,5,4’-trihydroxystilbene) for the prevention or as complementary therapies for eye diseases. Due to existing challenges in resveratrol delivery, our current approach is to load RV into polyelectrolyte multilayer microcapsules (PEM), whose release is controlled by a laser triggered procedure into retina pigmented epithelial cells. Mimicking the physiologic concentrations of resveratrol and hyperglycemia, which occurs in DR patients by using D407 cells, the unique in vitro experimental models for eye disease, we assess the therapeutic potential of RV-loaded PEM. The synthesis process of the complex is based on a simple Layer by layer assembly approach. During this procedure the capsules walls are decorated with collagen-coated gold nanoparticles for laser controlled release facilitation (figure on the left). The so obtained RV-loaded PEM were characterized using spectroscopic and microscopic methods. Next, the controlled release process of RV was optimized. The Raman signal from RV molecules is detected once the microcapsule is open by exposure to 532 or 633 nm laser light and the entrapped RV is released. Quantification of the RV released from microcapsule is assessed by HPLC-PDA. Based on the information obtained and on the specific formulas the RV loading content, RV entrapment efficiency and yield of production are determined for quantification of the therapeutic agent who reaches the target site.
Min Li
Huahai Pharmaceutical Co. Ltd, China
Title: Application of LC-MSn molecular fingerprinting in the rapid elucidation of drug impurities
Biography:
Min Li received his PhD in Organic Chemistry from Johns Hopkins University in 1991. Starting from 1994, he had worked for several multi-national pharmaceutical companies including Roche, Merck, and Schering-Plough/Merck with increasing responsibilities. Since 2014, he is Vice President for Analytical Operation at Huahai Pharmaceutical, a leading Chinese pharmaceutical company with operations in China and US. Dr. Li is a leading expert in drug degradation chemistry and he published a single-authored book, “Organic Chemistry of Drug Degradation” (by RSC) in 2012. He is the first/communicating author of 40 publications in organic, medicinal, bioconjugate, and analytical chemistry (including mass spectrometry).
Abstract:
ICH guidelines require that impurities of drug substances and products be identified and/or qualified once they exceed certain thresholds. The identification of these impurities, including process impurities and degradation products, can be quite challenging and time-consuming. In this presentation, the use of LC-MSn molecular fingerprinting in conjunction with knowledge of process and/or drug degradation chemistry will be shown to be a very effective way in the rapid elucidation of unknown drug impurities. MSn fragmentation patterns (n is typically 2 to 3) generated with suitable collision energy are highly reproducible on same type of MS instrument and collectively, they provide a unique molecular fingerprint for a particular impurity. With the relevant process chemistry (for process impurities) or drug degradation chemistry (for degradation products) in mind, comparison of the MSn fingerprint of an unknown impurity with those of known compounds, especially the isomers of the unknown impurity, can quickly yield critical structural information, which will greatly expedite the process of structure elucidation. This approach will be demonstrated through several case studies involving structurally diverse drug
Sreenivasa RJ
National Institute of Nutrition, India
Title: Determination of Heavy Metal Contents in Indian foods Using ICP-MS after Closed Vessel Microwave Digestion
Biography:
Sreenivasa RJ is an Asst. Director at National Institute of Nutrition, (ICMR), Hyderabad, India. He has completed his post graduation in Biochemistry and doctoral studies from Osmania University, Hyderabad. He has published several research papers in reputed peer reviewed journals and presented his research contributions in several National and International conferences and workshops. He holds life membership in professional scientific organization. He has been an invited distinguished speaker for many universities for discourse on heavy and trace metal toxicity, food composition and nutritional health. Presently he is working on, plant secondary metabolites and heavy metals in Indian foods using ICPMS. His core area of research also includes evaluation of carotenoid metabolism. He is a board member of NIN Scientist Association and Security and maintenance committee.
Abstract:
Heavy metals are natural components of the earth's crust and are widely used in agricultural, manufacturing and food/material processing industries. Among a variety of substances entering the soil, inland waters and the ocean as waste products, heavy metals like Nickel, Arsenic, Cadmium, Mercury and Lead are prominent. Heavy metal toxicity is a result of long term, low level exposure to pollutants through air, water and numerous consumer food products. In the field of food and nutrition the increasing levels of heavy metals in the environment, their entry into the food chain, and the overall health effects on people who consume green leafy vegetables (GLV) and fish are of major concern to researchers. In the present study the heavy metal content was analyzed from GLV and fish samples. Closed vessel microwave digestion system (CEM-MARS-USA).of food samples assisted by HNO3+ H2O2 were used to determine the heavy metals using ICPMS. CRM samples procured from NIST were used for method standardization and validation for linearity, repeatability, limits of detection (LOD) and limits of quantification (LOQ). Heavy metal measurements were performed using ICP-MS (PerkinElmer Élan 9000-USA). For better operating conditions the ICP-MS was adjusted to Nebulizer gas flow 0.91 L/min, Radio frequency (RF) 1200 W, Lens voltage 1.6 V, Cool gas 13.0 L/min, and Auxiliary gas 0.70 L/min. Microwave digestion followed by analysis by ICP-MS has been shown to be a simple, reliable method for the multi-element determination of heavy and trace metals in GLV and Fish samples. Results were in accordance with recommended human daily intake values except for arsenic, which were in agreement with WHO/FAO recommended values. Several reliable analytical methods like Colorimetric and Atomic Absorption Spectrophotometer are available for monitoring heavy metals in food samples, but ICP-MS is the most sophisticated and reliable technique, fast quantitative, high sensitivity, good precision and accuracy. Isotope ratio measurements were also possible using ICP-MS and interferences relatively few in number compared to other analytical techniques.
- Track 4: Advances in Sample Preparation Techniques
Session Introduction
Ruoting Zhan
Guangzhou University of Chinese Medicine, China
Title: Determination of mycotoxin contamination in Traditional Chinese Medicines by LC-MS-MS
Biography:
Professor Ruoting Zhan obtained his PhD in 2012 from Guangzhou University of Chinese Medicine, Guangzhou, China. He is currently the vice-director of Research Center of Chinese Medicine Resources Science and Engineering, Guangzhou University of Chinese Medicine. He specializes in studying resources of Chinese medicinal materials. His researches focus on authenticating the medicinal herbs in South China area, which includes resources survey on the botanical origins, identification of the commercial products, the pharmacognosy studies on different species, etc. His most recent work emphasizes the importance of the sustainable development and utilization of traditional Chinese medicine resources. He has published more than 30 papers in reputed journals.
Abstract:
Mycotoxins are toxic secondary metabolites from molds. Presently, liquid chromatography-tandem mass spectrometry (LC-MS-MS) has been widely used to evaluate mycotoxin contaminations in food and feed. However, application of LC-MS-MS to Traditional Chinese Medicines (TCMs) is difficult，because their complex matrices can cause matrix effect. Routinely, some purification procedures like immunoaffinity columns (IAC) should be included before injection to improve the accuracy and protect mass spectrometer. However, the high costs of columns restrict their application for large-scale screening. Therefore, the objective of our study is to develop a simple, rapid and cost-effective LC-MS-MS method for simultaneous determination of aflatoxins (AFB1, B2, G1 and G2) and sterigmatocystin (ST) in TCMs. The method is based on single extraction with (84/16, v/v) acetonitrile-water then analysis of the diluted crude extract without further clean-up. The chromatographic separation was achieved on a C18 column, with a mobile phase gradient prepared from aqueous 4 mmol L-1 ammonium acetate-0.1% formic acid and methanol. Quantification of the analytes was by selective reaction monitoring (SRM) on a triple quadrupole mass spectrometer in positive ionization mode. Special focus was on investigating and reducing matrix effects to improve accuracy. The established method was validated by determination of linearity, sensitivity, extraction recovery and precision. Comparison with IAC pretreatment demonstrated both methods are comparable in respect of the recovery of aflatoxins. Finally, the validated method was used to evaluate mycotoxin contamination in total 294 samples of 30 TCMs collected from local hospitals and pharmacies and the results obtained were analyzed.
Biography:
Peter Mikuš has completed his PhD at the age of 30 years from Comenius University (Slovakia). He is researcher, university teacher, associated professor, and director of the Toxicological and Antidoping Center at the Faculty of Pharmacy Comenius University in Bratislava (FPCU) as well as head of the Department of Pharmaceutical Analysis and Nuclear Pharmacy FPCU. A research team of P.M. is focused on the development, validation and application of advanced hyphenated analytical methods, based on a combination of 2D-separation and spectral (UV-VIS, MS/MS)
Abstract:
A new highly advanced multidimensional analytical approach for the ultra-trace determination of target chiral compounds in unpretreated complex real samples was developed in this work. The proposed analytical system provided high orthogonality due to on-line combination of three different methods (separation mechanisms), i.e. (i) isotachophoresis (ITP), (ii) chiral capillary zone electrophoresis (chiral CZE) and (iii) triple quadrupole mass spectrometry (QqQ MS). The ITP step, performed in a large bore capillary (800 ïm), was utilized for the effective sample pretreatment (preconcentration and matrix clean-up) in a large injection volume (1-10 ïL) enabling to obtain as low as ca. 80 pg.mL-1 limits of detection for the target enantiomers in urine matrices. In the chiral CZE step, the different chiral selectors (neutral, ionizable, and permanently charged cyclodextrins) and buffer systems were tested in terms of enantioselectivity and influence on the MS detection response. The performance parameters of the optimized ITP – chiral CZE-QqQ MS method were evaluated according to the FDA guidance for bioanalytical method validation. Successful validation and application (enantioselective monitoring of renally eliminated pheniramine and its metabolite in human urine) highlighted great potential of this chiral approach in advanced enantioselective biomedical applications.
Biography:
he is of Head of the IRBM team for Merck Research Laboratory inter-site Harmonization for the Bioanalytical Procedures. At present consultant at IRBM Science Park. Consultant at AlfaWassermann spa. Consultant at Università del Sacro Cuore A.Gemelli. Autor of more than 40 publications and seven patent
Abstract:
This work describes a simple, sensitive and rapid liquid chromatography-high resolution mass spectrometry method for quantitation of perhexiline and the simultaneous detection of perhexiline metabolites in C57b1/6 mice plasma. Only 5 µL of plasma was used for analysis. Pretreatment was limited to a 100-fold dilution (‘dilute-and-shoot’). The analyte was detected by high resolution mass spectrometry (Orbitrap Technology). Three scan events were performed over the entire chromatogram. Targeted single ion monitoring with data dependent acquisition was employed for perhexilline quantitation and confirmation, while full scan mode was used to perform untargeted detection of perhexiline phase I and phase II circulating metabolites. The calibration curve was linear (r2 = 0.990) ranging from 0.305 ng/mL (LLOQ) TO 10000 ng/mL. Matrix effect was limited to 6.1%. The method was applied to a pharmacokinetic study of perhexiline in mouse plasma and the results obtained were compared to a standard sample preparation method based on protein precipitation and liquid chromatography-tandem mass spectrometry (MRM mode) detection. The new approach provided comparable results in terms of pharmacokinetics parameters estimate with a high sensitivity, additional information on perhexiline circulating metabolites and a low consumption of biological sample. The combination of the ‘dilute-and-shoot’ approach together with HRMS targeted and untargeted detection represents a suitable alternative to classic bioanalytical approaches in preclinical research.
K. B. Rameshkumar
Jawaharlal Nehru Tropical Botanic Garden and Research Institute, India
Title: Phytochemical Diversity of the Flora of the Western Ghats-Mass Spectrometric Evaluation
Biography:
Dr. K. B. Rameshkumar took his Ph.D in chemistry from University of Kerala, Thiruvananthapuram in 2008, and has been working in the Phytochemistry Division of Jawaharlal Nehru Tropical Botanic Garden and Research Institute (JNTBGRI), Thiruvananthapuram as a scientific staff since 1998. He had also worked in Sultan Quaboos University, Muscat, Oman as a ‘Visiting Scientist’ during 2011. He had worked on the phytochemistry of several hitherto uninvestigated endemic plants of the region and received the prestigious ‘Young Scientist’ award by Kerala Science Congress, K.S.Manilal Award by the Indian Association for Angiosperm Taxonomy and P.D. Sethi Award for the best paper in HPTLC. He had published more than 30 research papers in peer reviewed journals.
Abstract:
The Western Ghats, one among the 34 global biodiversity hotspots, harbor nearly 7500 flowering plants, of which 1250 are endemic to the region. The present study reports the phytochemical diversity of selected plants of the Western Ghats through hyphenated mass spectrometry techniques. LC-MS studies of 13 Garcinia species revealed distribution of bioactive constituents such as biflavonoids, xanthones and acids, while GC-MS studies reported the genus as rich source of volatile terpenoids and aliphatic compounds. Distribution of piperamides, phenolics and terpenoids in the fruits and leaves of ten Piper species were studied by various LC-MS methods. Direct analysis in real time mass spectrometry (DART-MS) has been found as a rapid and reliable tool for demarcation of the medicinal herbs Piper nigrum, P. longum and P. chaba. Also the chemotaxonomic marker compounds were determined for Piper species based on leaf volatile chemical studies through GC-MS. Several new natural sources of aroma compounds such as camphor and linalool were discovered through the GC-MS studies of Cinnamomum species from the Western Ghats. The hepatotoxic phenyl propanoid coumarin in cinnamon samples (C. verum) were found within the tolerable daily intake limit by UHPLC-ESI-Qqq-LIT-MS method. HPLC-QTOF-MS studies on the distribution of phorbol esters in Euphorbia species revealed that the toxic phorbol ester TPA was absent, while the cytotoxic ester prostatin was present in the all the 13 Euphorbia species studied. More than 80% of the endemic flowering plants of the Western Ghats region are hitherto uninvestigated for their chemical constituents and the present study highlights the application of various hyphenated MS techniques in chemical profiling of the unexplored flora.
Amarjit Singh Sarpal
Indian Oil Corporation Ltd., India
Title: Mass Spectrometric techniques for investigation of biodiesel potential of Microalgae biomass
Biography:
Dr. A. S. Sarpal completed his Ph.D degree in analytical chemistry in 1980 from GNDU, Amritsar, India and worked in the IndianOil Corporation Ltd (R&D)., India from 1977 to 2011. He has 40 years experience in the characterization of petroleum and related products, bio-fuels, polymers and catalyst by the applications of analytical techniques such as FT NMR, FTIR, MS, GC-MS etc. and published 135 research papers. He has also worked at University of Illinois in 2011on a project on biofuels from algae biomasses and capacity of team leader at INMTRO), Rio de Janeiro, Brazil from April 2012 to October 2015.
Abstract:
Microalgae oils have emerged as potential sources of nutrition, energy and oil besides useful bi-products, such as nutraceuticals, antioxidants and high performance lubricant additives. Microalgae are unicellular photosynthetic organisms with 5-50 µm in size that require primarily three components to produce biomass, i.e., water, CO2 and sunlight with relatively higher photosynthetic efficiency against terrestrial plants. Microalgae biomass is comprised of neutral (tri acyl glycerides, TAG; free fatty acids, FFA) and polar lipids (glyceroglyco/phospholipids) besides specific molecules with high industrial potentials. Neutral lipids are imortant feedstocks of biodiesel due to their similarity with regular vegetable crops with regard to saturated and unsaturated fatty acid profile (C14 to C22) including C20:3, C20:5 (EPA) and C22:6 (DHA). The presentation will highlight the role of Mass spectrometric techniques (ESI-MS;TOF, QMS; GC-MS) for determination of composition of microalgae oils generated from biomasses of Dunalialla salina, Chlorella vulgaris, Spirulina platensis and Scemedesmus oblique species cultivated on a lab scales under different growth parameters in fresh and industrial waste water with an emphasis on exploring the biodiesel and PUFAs potential.
Elliot R. Bernstein
Colorado State University, USA
Title: Euv laser mass spectrometry and photoelectron spectroscopy of mass selected neutral clusters and molecules
Biography:
Prof. Bernstein received his Ph.D. degree from Caltech and was a post doctoral fellow at the University of Chicago. He has been at CSU since 1975 where he has studied molecular crystal vibrational and electronic excitons and phase transitions, cryogenic liquids, and gas phase clusters with a central focus on intermolecular interactions. Recently his research has focused on chemical reactions of neutral and ionic clusters. This latter research area has involved catalytic and photocatalytic cluster systems, solute/solvent systems, the reactions of ionized molecules and clusters, and initial steps in the release of stored energy molecular species.
Abstract:
A long standing set of goals for studies of systems of inhomogeneous, neutral clusters (e.g., MmXn or (molecule)p) has been to mass sort and select them individually for determination of physical and chemical properties of each neutral cluster by spectroscopic techniques. We have constructed appropriate instrumentation to achieve these important goals employing photoelectron spectroscopy (PES), driven by both visible (for MmXn -) and EUV (for MmXn0) radiation. Our 26.5 eV/photon EUV laser can ionize any neutral cluster or molecule (EUV PES) that can be identified and isolated. The algorithm includes the following steps: 1. generation of cluster negative ions in a laser ablation supersonic source with the addition, as required, of low energy electrons from a Y2O3 disk; 2. separation of these anionic clusters in a reflectron time of flight mass spectrometer (RTOFMS); 3. selection and slowing of specific, chosen clusters in a mass gate/momentum deceleration stage; 4. threshold photo-detachment of the sorted and selected negative ion clusters with a tunable VIS/UV laser to generate neutral, isolated clusters; and 5. EUV PES of these neutral clusters. Such studies generate vibrational and structural information on the ground states of the neutral clusters (through VIS/UV PES), and information on the ion states of the clusters (through EUV PES). The presentation will include PES results on various metal oxides, sulfides, and other cluster systems and molecules.
Seitkerim B. Bimurzaev
Almaty University of Power Engineering and Telecommunications, Kazakhstan
Title: A TOF mass-spectrometer with eliminated TOF chromatic aberration of the first order
Biography:
S.B. Bimurzaev is doctor of Physics and Mathematics, Professor of the Computer Technology Department of the Almaty University of Power Engineering and Telecommunications (Kazakhstan). He has published more than 30 papers in reputed journals such as «Nucl. Instr. Meth. Phys. Res. A» (Elsevier), «Journal of Electron Microscopy» (Elsevier), «Int. Journal of Mass Spectrometry» (Elsevier), «Technical Physics Letters» (Springer) as well as in « Sov. Phys.: Journal Tekhnicheskoi Fiziki», «Sov. Phys.: Radiotekhnika and Electronika», etc.
Abstract:
As is known, high resolution in modern TOF mass spectrometers of mass reflectron type is achieved by the use of special devices – ion reflectors of various modifications – in their ion optical systems [1,2]. TOF mass spectrometers without ion reflectors, for example [3], have relatively low resolution. The main reason limiting the resolving power of TOF mass spectrometers of any design is TOF chromatic aberration of the first order caused by the initial energy spread of ions in the packets generated by the ion source. In this paper we investigate the possibility of eliminating TOF chromatic aberration of the first order with the help of ion accelerator with an inhomogeneous electrostatic field. It is shown that for a certain choice of the field distribution of such an accelerator, along with the elimination of TOF chromatic aberration of the first order, it is possible to obtain a high-quality TOF focusing ion packets in the plane of the detector simultaneously with spatial focusing. The author also considers schemes of TOF mass spectrometers of simple construction (with no ion reflector) with high resolution and sensitivity.
Achille Cappiello
University of Urbino, Italy
Title: Advancements in LC Direct-EIMS: State of the Art and Applications
Biography:
Achille Cappiello is a professor of Analytical Chemistry at the University of Urbino in Italy, He studied at the University of Rome La Sapienza and after a two-year appointment as Post Doctoral Associate at the Massachusetts Institute of Technology, supervised by Professor Klaus Biemann, he began to engage in the field of LC-MS. Professor Cappiello is the director of several projects dealing with LC-MS instrument development and design with some of the major manufacturers. He published more than 100 scientific articles with an international circulation including the top journals in the field such as “Analytical Chemistry†and “Mass Spectrometry Reviewsâ€.
Abstract:
Mass spectrometry (MS) is renowned for its impressive identification potential especially when it is coupled to a separation technique such as liquid chromatography (LC-MS) or gas chromatography (GC-MS). Direct-EI LC-MS interface, developed in our lab, is a powerful technique that combines, in a single instrument, the identification advantages of library searchable, electron ionization (EI) spectra with the separation power of an LC column, without the drawbacks of matrix effects or the polarity limitations typical of electrospray ionization. Non target analyses are of increasing importance in food safety, environmental, forensic and many other applications where the complexity of the matrix is a troubling factor. The advantage of EI for tentative identification is unparalleled. Expansion of EI fragmentation to a wider variety of molecules in a liquid phase provides an attractive alternative to identification and offers a complementary technique to high-resolution/high-mass accuracy LC-MS instrumentation and atmospheric pressure techniques. The simple interfacing process make it compatible with any LC separation technique and with any solvent combination, including non-volatile buffers and UHPLC methods. The gas-phase ionization process and the negligible matrix effects allows direct injections of samples even without column separations. In this presentation, the basic principles of compound identification in various applications and the advantages offered by an MS/MS analyzer in combination to HPLC and UHPLC separations will be presented. New instrumental developments involving the use of ceramic coatings and membrane probes for additional sensitivity and flexibility will be also shown.
Daniele B. Rodrigues
University of Campinas, Brazil
Title: Further insights on the carotenoid profile of Tagetes patula flowers
Biography:
Daniele Bobrowski Rodrigues has a Bachelor’s degree in Pharmacy and a Master’s degree in Food Science and Technology. She is a PhD student in the Food Science Department at University of Campinas-Brazil. The main themes of her research are extration and identification of pigments by HPLC-DAD-MS/MS, antioxidant capacity and carotenoid bioaccessibility.
Abstract:
Identification of carotenoid esters is a challenge and much more difficult and laborious than that of free carotenoids in saponified extracts since the number of possible carotenoid esters found in plant materials is quite high. Xanthophylls can have 1 or 2 hydroxyl groups in the molecule which can be acylated or not with several fatty acids. We carried out a comprehensive identification of the carotenoids from marigold petals by LC-APCI-MS/MS and identified 56 carotenoids: 6 free carotenoids, 20 monoesters and 30 diesters. This was the first time that esters of auroxanthin, zeinoxanthin and ï¢-cryptoxanthin were identified in marigold petals, while lutein esters were the major carotenoids, as expected. Differentiation between lutein mono- or heterodiester isomers was based on the different intensities of the ion fragments in the MS spectrum due to the loss of the substituent attached to the ï¥-ring (more stable and more intense m/z signal) and the one attached to the β-ring. Moreover, the heterodiester with the fatty acid of the highest molecular mass acylated at the 3’-O-position eluted before the correspondent 3-O-regioisomer. For instance, both lutein 3-O-laurate-3’-O-myristate and lutein 3-O-myristate-3’-O-laurate presented in their MS spectra the fragment ions at m/z 733 [M+H-myristic acid]+ and m/z 761 [M+H-lauric acid]; however, in the first diester MS spectrum, the fragment ion at m/z 733 was more intense than at m/z 761 and the contrary occured for the other isomer.
Regina Stabbert
Philip Morris Products S.A., Philip Morris International R&D, Switzerland
Title: Heterocyclic aromatic amines and their contribution to the bacterial mutagenicity of the particulate phase of cigarette smoke
Biography:
Dr. Regina Stabbert received her Ph.D. in chemistry in 1992 from the University of Cologne. She is Principal Scientist working in the team Product Stewardship at Philip Morris International, RRP and is a manager with more than twenty years of experience in cigarette smoke chemistry, toxicology and aerosol science. She has published more than 15 papers in peer-reviewed journals.
Abstract:
Heterocyclic aromatic amines (HAAs) rank among the strongest known mutagens. Approximately 30 HAAs have been found in cooked foods (broiled, fried, and grilled) and several HAAs have been characterized as animal carcinogens. Nine HAAs have also been reported to be constituents of cigarette smoke (CS) raising concerns that HAAs might contribute significantly to the known carcinogenicity of CS. An improved method for the quantification of HAAs in the total particulate matter (TPM) of CS is reported allowing detection and quantification of 8 HAAs in a single run. The mutagenic potency of these HAAs and that of TPM from the reference cigarette 2R4F was determined in the Salmonella Reverse Mutation Assay (Ames Assay) with tester strain TA98 and a metabolic activation system. The 8 HAAs, when applied together in the Ames assay, showed a clear sub-additive response; that means that mixes of the 8 HAAs gave rise to responses that were distinctly below that expected under the assumption of additivity for the single HAAs. Likewise, the combination of HAAs and TPM, if at all, gave rise to a slight sub-additive response . In both cases, however, the sub-additive response in the Ames assay was observed at HAA doses that are far above the amounts found in CS. The contribution of the individual HAAs to the total mutagenic activity of TPM was calculated and experimentally confirmed to be approximately 1% of the total mutagenic activity. Thus, HAAs do not contribute significantly to the bacterial in vitro mutagenicity of TPM in CS.
Eduard Rogatsky
Wadsworth Center, USA
Title: 25 Hydroxy Vitamin D quantitative analysis by LC/MS – assay standardization
Biography:
Dr Rogatsky is a senior faculty member at Albert Einstein College of Medicine (NY, Bronx) and director of mass spectrometry at Biomarker Analytical Resource Core as part of the Harold and Muriel Block Institute for Clinical and Translational Research at Einstein and Montefiore . He has worked in the field of chromatography more than 20 years. Since 2001 his work has been within the service of the field of clinical mass spectrometry. During last 10 years (from 2004) Dr Rogatsky published 25 scientific papers in per-reviewed journals (mostly as the first author) and presented over 50 posters and lectures. Currently Dr Rogatsky serve as the Editor-in-Chief for the Journal of Chromatography and Separation Techniques (OMICS publishing group). Eduard Rogatsky completed his M.Sc (physical chemistry) in Belarus State University (former USSR) in 1990. In 1998 has completed PhD in bioanalytical chemistry (Bar-Ilan University, Israel). At the end of 1999 he started post-doctorate at Albert Einstein College of medicine and since 2001 joined faculty. Currently his holds a title of Research Associate professor of Medicine.
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 droplets evaporation region can vary with mass spectrometer hardware design, ratio between intact [M+H]+ and dehydrated precursor can be unpredictable. We also noticed that degree of dehydration is concentration-dependent. Chromatographic separation between analyte and its deuterated internal standard might cause different levels of analyte and internal standard dehydration and resulted in quantitative error.
Rodrigo De Paula
, University of Aveiro, Portugal
Title: The strategies in development of synthetic metalloporphyrins include high selective oxidation
Biography:
Rodrigo De Paula obtained his degree in Chemistry in from University Federal Uberlândia (Minas Gerais/Brazil). At the same university, he obtained his MSc in Chemistry in 2003. In the same year, he was employed as Global Coordinator Development at Unilever Brasil and in 2005 he has moved to Portugal, where began his PhD in Aveiro (2005) on synthesis and catalytic studies of porphyrins and its metallocomplexes. In 2010 he came back to Brazil as Adjunct Professor of Chemistry at University Federal of Recôncavo Bahia. He has published works in two main lines, photophysics and photochemistry of dyes and pigments as well as in metalloporphyrins synthesis and catalysis. He is Ad-Hoc referee of some important Journals as Organic Chemistry, Dalton Transactions, RSC Advances and Catalysis Science & Technology. He is the Head of Catalysis Research (GPCat) and its research on synthesis and structrural, spectroscopic and catalytic characterization of porphyrins and phthalocyanines if sponsored by national research agency CNPq.
Abstract:
Porphyrins and metalloporphyrins are natural occurring compounds. The synthetic derivatives are used in many fields of science such as photodynamic therapy, laser dyes, photocatalysts, biomimetic models for olefins and drugs oxidation. Concerning olefins oxidation, the synthetic metalloporphyrins (Iron and Manganese complexes) are able to mimic the Cytochrome P450 function. This enzyme is Fe(III)-heme surrounded by a bulky protein. The strategies in development of synthetic metalloporphyrins include high selective oxidation, robustness and the reuse. To achieve that characteristic, it is usual to attach metalloporphyrin onto a solid support. This work report some strategies developed in preparing efficient and robust manganeseporphyrins and its use as catalyst oxidation as well as some important kinetic and thermodynamics aspects into reaction.
Suchi Chawla
Anand Agricultural University, India
Title: Matrix Effects of different food matrices in quantitative analysis of pesticides in chromatography coupled to tandem mass spectrometry
Biography:
Dr (Mrs) Suchi Chawla, completed PhD from Jawaharlal Nehru University, New Delhi, India. I have been working as Research Associate for last 8 years in Pesticide Residue Laboratory at Anand Agricultural University, Anand, Gujarat, India. Present job involves (i) Method development and validation for multiresidue analysis in different food matrices on LC-MS/MS and GC-MS/MS. (ii) Quality control and assurance activities of the laboratory as per ISO 17025:2005. (iii) Conducting sponsored trials as per GAP to get residue and persistence data for MRL fixations (iv) Monitoring of pesticide residue in food at national level. I have 4 publications in international journals and 1 in national journal. Earlier I have worked with Indian Journal of Medical Research in publication division.
Abstract:
Injudicious use of pesticides has lead to increase in the pesticide levels in the environment and food. This increase may pose many health hazards to humans and can affect the animals and various ecosystems adversely. Consumption of pesticide contaminated food via daily diet is a major source of exposure to pesticides. It is necessary to monitor various pesticide residues in food commodities. Most of the pesticide analysis is done by multi-residue analysis methods as it is less time consuming as compared to single residue methods. Availability of more sensitive chromatographic techniques and tandem mass spectrometry coupled to chromatography (GC, GC-MS/MS, LC-MS/MS) has improved the selectivity and sensitivity of pesticide analysis and their identification and quantification in agricultural products. The quantitation by chromatography and/or chromatography coupled to mass spectrometry is affected by matrix in the multi-residue analysis leading to suppressed or enhanced results due to matrix effects. The matrix effect of a compound is the change in signal in a solvent solution compared with signal in matrix due to presence of co-extracted compounds, or changes in eluent properties, such as surface tension, viscosity, volatility – all factors known to affect the ionization process. The suggested ways to reduce the matrix effect are (i) sample clean-up (ii) introduction of additives into the mobile (iii) internal standards or isotopically labeled external standards prepared in sample matrix, (iv) dilution of matrix to reduce matrix effects. But it is difficult to obtain a blank matrix for every sample type and will immensely decrease the number of analysis per day. In the present study, we investigated matrix interferences by QuEChERS sample preparation to establish whether (i) dilution of matrix can eliminate the need of matrix standards in LC-MS/MS (ii) the commodities can be grouped together, thus reducing the number of matrices needed to make the calibration standards. Matrix effects were studied by comparing of slopes of calibration curves of matrix (diluted with mobile phase) and solvent based standards. Present study showed that matrix effects were dependent on both, nature of commodity as well as analyte. Maximum matrix effect variability was observed in capsicum. Most of the pesticides showed ion suppression in tomato, capsicum and cumin matrices. In brinjal matrix most of the pesticides showed slight ion enhancement, though the extent of ion enhancement was very less. Due to similar nature of matrix effect of tomato and capsicum these two commodities can be grouped together. Cumin matrix was most difficult to analyse. As 4X dilution did not completely eliminated matrix effects a comparison was made between 10X diluted matrices of cucumber and brinjal. Recovery study was also undertaken in brinjal was done and compared with both the matrices. Matrix effect did not vary much between the two matrices. Analyte variability was also less. Most of the pesticides showed recoveries in acceptable range of 70-130% with calibration curves from both matrices. To compensate for matrix effect it is suggested that (i) tomato and capsicum matrix which show similar trend can be grouped together, (ii) 10 times diluted cucumber matrix can be used to prepare calibration curves for quantitation of pesticides on LC-MS/MS from different fruiting and cucurbits vegetable matrices with LOQ of 0.05 mg kg-1. Further studies can be done with other vegetables like chillies, onion, garlic, spinach, fruits, cereals, pulses and spices to find candidate matrix for quantitation. To understand the effect of mode of ionization on matrix effect, the matrix effects in LC-MS/MS and GC-MS/MS were compared.
Pim de Voogt
University of Amsterdam and KWR Watercycle Research Institute, The Netherlands
Title: Determination of drugs of abuse and related compounds in sewage water by liquid chromatography coupled to high resolution mass spectrometry
Biography:
Pim de Voogt has completed his PhD from Vrije Universiteit in Amsterdam (The Netherlands) in 1990. He is professor of environmental chemistry and chair of the department of aquatic environmental ecology at the University of Amsterdam, and also principal scientist at KWR in Nieuwegein, The Netherlands. He has published more than 200 papers in reputed journals and books and he is serving as editor-in-chief of Reviews of Environmental Contamination and Toxicology.
Abstract:
Wastewater-based epidemiology is a new field of research in which analysis of concentrations of biomarkers of drugs in domestic wastewaters is used to back calculate human consumption of the drugs. Liquid chromatography coupled to mass spectrometry has been successfully used to estimate and compare consumption of illicit drugs and counterfeit medicines in European city populations. Apart from target screening of classical drugs like cocaine and amphetamine the presence of new psychoactive substances (NPS, designer drugs) can be screened in wastewater, e.g., during festivals and other big events, using high resolution MS. For this purpose, a qualitative screening method based on liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOF MS) was applied for the detection and identification of stimulant-type NPS in wastewater. Eight 24-h composite influent wastewater samples were collected at the wastewater treatment plant (WWTP) serving the catchment area of Amsterdam in 2012 and 2014, during two events that each brought 300,000 visitors to the city.
- Track 10: Maintenance, Troubleshooting, Data Analysis and Experimentation in Mass Spectrometry
- Track 1: Applications of Mass Spectrometry Track 3: New Approaches in Mass Spectrometry Track 6: Mass spectrometry Imaging Track 8: Ionization Techniques
Chair
Elliot R Bernstein
Colorado State University, USA
Co-Chair
Ioana NUTA
Univ. Grenoble Alpes, SIMAP, France
Session Introduction
Yujun Shi
University of Calgary, Canada
Title: Development of Laser Ionization Methods and Its Application in the Investigation of Chemical Vapor Deposition Chemistry
Biography:
Yujun Shi obtained her PhD in Chemistry from the University of Western Ontraio (now Western University) in Canada. She did postdoctoal work at the National Research Council (NRC) of Canada in Ottawa on an NSERC Visiting Fellowship. She is currently an Associate Professor in the Department of Chemistry at the University of Calgary. She has published more than 50 papers in referred journals and has been serving as an editorial board member for Canadian Journal of Chemistry (Associate Editor) and Frontiers in Physical Chemistry Chemical Physics.
Abstract:
Laser ionization mass spectrometry is a powerful diagnostic tool for a variety of applications including biomedical research, drug delivery, atmospheric chemistry, and chemical vapor deposition chemistry. Our laboratory has developed several complimentary ionization methods, including the non-resonant single photon ionization (SPI) with a vacuum ultraviolet (VUV) laser radiation of 118 nm (10.5 eV) and the laser induced electron ionization (LIEI) for the diagnosis of the decomposition chemistry of precursor molecules in the process of hot wire chemical vapor deposition (HWCVD). In this work, results from our investigation on the decomposition of orgaosilicon molecules on heated tungsten and tantalum wires will be reviewed. A common decomposition pathway of forming methyl radicals has been demonstrated for methy-substituted silane molecules and methyl-substituted four-membered-ring silacyclobutane compounds. The formation mechanism and the activation energy for this pathway will be presented in this work. For the four-membered-ring organosilicon precursors, additional decomposition channels were also found from the ring-opening reactions to form alkene and silenes. The investigation of the decomposition chemistry of organosilicon molecules on heated metal wires provides helpful and important insights in understanding the secondary gas-phase reactions in HWCVD processes and also the surface reactions on the metal catalysts.
Andrew Ewing
University of Gothenburg and Chalmers University of Technology, Sweden
Title: Mass Spectrometry Imaging in Flies, Cells, and Vesicles
Biography:
Andrew Ewing received a PhD from Indiana University. After 25 years at Penn State University, he is now Professor at Chalmers University and University of Gothenburg, Sweden. His 300 publications have been cited over 15000 times with an H-index of 69. He has recently received the Norblad-Ekstrand Medal of the Swedish Chemical Society (2014), and the Pittsburgh Conference Award in Analytical Chemistry (2015). He is an Honorary Professor at Nanjing and Beijing Universities of Science and Technology. He is a member of the Royal Swedish Academy of Sciences (from 2012) and the Gothenburg Academy of Arts and Sciences (2013).
Abstract:
We have been developing mass spectrometry imaging methods to study the process of neurocommunication at the system and cellular level. We focus on PC12 cells as a model of exocytosis and the fly model (Drosophila melanogaster) providing a unique system to examine neurotransmitter release and drug dependence mechanisms in a small, but complete system. Mass spectrometry imaging with ion beams allows spatial resolution of a few micrometers down to 40 nanometers in favorable cases. We have been using secondary ion mass spectrometry (SIMS) with a unique 40-kV argon cluster ion source and the NanoSIMS to measure the lipids across the fly brain and catecholamine in nanometer vesicles, respectively. Here, we have focused on the effect of the drug, methylphenidate, on lipid composition in the brain and find that it varies in a way that might affect learning and memory. We have also used NanoSIMS to measure transmitter in subregions of nanometer vesicles. Combined with other new methods to measure the content of the interior of vesicles, we have begun to investigate the details and implications of open and closed exocytosis on regulation of how the brain works.
Libong Danielle
Paris South University, France
Title: Identification of lipid biomarkers of atopic skin by normal phase chromatography/high resolution mass spectrometry
Biography:
Danielle Libong has completed her PhD at the age of 28 years from Ecole Polytechnique of Palaiseau (France). She is senior lecturer in the Interdisciplinary Academic Unit Lip(Sys)² Lipids (Analytical and Biological Systems) of the School of Pharmacy of Paris-South University. Lip(Sys)² is a research team in analytical chemistry with an important experience in lipid analysis by chromatographic, hyphenated methods and optical spectroscopy. She has published more than 25 papers in reputed journals.
Abstract:
The strategy used to determine the lipid biomarkers involve lipidomics analysis of normal and atopic skin. In the case of skin called atopic, permeability of the epidermis or the stratum corneum is a problem. Stratum Corneum has been described as a type of “brick and mortar”, in which the corneocytes are bricks. Extracellular spaces are filled with lipids that are divided into three main classes: ceramides, fatty acids and cholesterol. Ceramides constitute a major class of lipids and are composed of a sphingoid and a fatty acid moiety. They play a major role in skin permeability. Their organization is essential to maintenance of the barrier function. Lipidomics offers a unique opportunity to analyze the complex role of lipids in cellular processes. The first step focuses on the development of a method for separation and detection of lipid classes molecular species using (Normal Phase) LC/ MS in high resolution. Different ionization modes have been tested between APPI and APCI. The main goal is to collect information from all detected peaks, the profile of the sample provides the relative distribution of species and the molecular mass of each. Statistical comparison of the profiles obtained for several samples of both populations reveal the characteristic signals of over / under expressed signals in each group. In the end we characterize and identify biomarker by MS/MS.
Ioana NUTA
Univ. Grenoble Alpes, SIMAP, France
Title: Mass spectrometry with Knudsen cell applied to the investigation of organometallic precursors vapors
Biography:
Ioana NUTA received her Ph.D from Orleans’s University (France) in 2005 working on fluoride molten salts baths at high temperature by NMR spectroscopy in CNRS-CEMHTI Laboratory. She then completed a post-doctoral fellowship at ENSI Caen (2006) in CNRS-LCS Laboratory and at BRUKER (Germany) in department of NMR probe devellopement for catalytic studies. Since 2007, she joined CNRS- SIMAP Laboratory as reasercher in “Thermodynamic and Process Optimization” team where she has in charge the themodynamic studies on gaseous phase using Knudsen effusion Mass Spectrometery. Current studies focus on thermodynamics of organometallics, molten salts and oxides.
Abstract:
A special Knudsen cell reactor coupled to a mass spectrometer, was specifically designed for the study of organometallic precursors. This reactor is built as tandem cells: an evaporation cell and a cracking cell. This reactor tries to simulate the conditions found in the bubbler of the ALD system by the evaporation cell and in the hot reaction zone of ALD by the cracking cell. The first stage reactor - an evaporation cell - provides an input saturated vapor flow operating from room temperature to 333 K. The second stage cell, named cracking cell, operated from 333 to 723K in the present study. During experiments, the effusion orifice is externally opened for direct mass spectrometric measurements of saturated vapour pressures. The device has been tested using the well-known mercury system. The thermal cracking of the gaseous precursor pentakis dimethylamino tantalum (PDMAT), generally adopted in the ALD/CVD TaN deposition processes, has been studied in the temperature range from 343 to 723K. Experiments showed the apparition of many gaseous species when cracking temperature increased and in particular the dimethylamine, HNC2H6 (g), corresponding to the saturated organic branches of PDMAT. Decomposition products of the HNC2H6 branch were observed at relatively high temperature, namely above 633K. This gas phase study - as for the saturated preceding one- shows the presence of oxygen containing molecules in PDMAT cracked vapor. Thus it allows explaining the systematic presence of oxygen contamination in the deposited TaN films observed in ALD/CVD industrial processes.
Stefanie Maedler
Ontario Ministry of the Environment and Climate Change, Canada
Title: Ultra-trace level speciated isotope dilution measurement of Cr(VI) using ion chromatography tandem mass spectrometry in environmental waters
Biography:
Stefanie Maedler received her Ph.D. in analytical chemistry in 2011 from the Swiss Federal Institute of Technology in Zurich, Switzerland (ETHZ). After 2 postdoctoral fellowships at York University and University of Toronto, Canada, Dr. Maedler joined the Ontario Ministry of Environment and Climate Change as a Development Scientist. She has 10 years of expertise in the use of mass spectrometry (MS) for biological and environmental applications and is currently focusing on the speciation of inorganic analytes and understanding their toxicological effects on fish through MS-based metabolomics approaches.
Abstract:
The reliable analysis of highly toxic hexavalent chromium, Cr(VI), at ultra-trace levels remains challenging, given its easy conversion to non-toxic trivalent chromium. The new approach demonstrates a novel analytical method to quantify Cr(VI) at low ng/L concentration levels in environmental water samples by using speciated isotope dilution (SID) analysis and double-spiking with Cr(III) and Cr(VI) enriched for different isotopes. Ion chromatography tandem mass spectrometry (IC-MS/MS) was used for the analysis of Cr(VI) as HCrO4- → CrO3-. While following a classical linear multipoint calibration curve a method detection limit (MDL) of 7 ng/L Cr(VI) was achieved, the modified SID-MS method adapted from U.S. EPA 6800 allowed for the quantification of Cr(VI) with an MDL of 2 ng/L and provided results corrected for Cr(VI) loss occurred after sample collection. The adapted SID-MS approach proved to yield more accurate and precise results than the multipoint calibration method, allowed for compensation of Cr(VI) reduction during sample transportation and storage while eliminating the need for frequent external calibration. The SID approach permitted continuous sample analysis for several days without the need for recalibration. This new developed IC-MS/MS method represents an alternative to the routinely used inductively-coupled plasma (ICP) instrumentation, IC-ICP-MS, and offers several advantages over detection with ICP-MS for Cr(VI), such as the absence of polyatomic interferences of 52Cr formed in the ICP (36Ar16O, 40Ar12C, 35Cl16OH, and 37Cl14NH) that require the use of dynamic reaction/collision cells or high-resolution double-focusing sector field instruments.
Nhu Phan
University of Gothenburg, Gothenburg, Sweden
Title: Lipid imaging of invertebrate model systems by secondary ion mass spectrometry
Biography:
Nhu Phan has been working on her PhD at the University of Gothenburg, Sweden. Her research is about developing mass spectrometry imaging methods for biological applications on different biological models, from single cell imaging to invertebrate models particularly C. elegans and Drosophila. One of her main researches is studying the effects of administrated stimulant drugs on the neurochemistry of Drosophila brain. The significant finding in this study is that the stimulant drug induces lipid structure of the brain and that the brain lipids could closely relate to learning and memory.
Abstract:
Drosophila melanogaster and C. elegans (flies and worms, respectively) are common biological model systems, which have relatively simple anatomy and behavior but possess highly conserved molecular and cellular processes compared to humans. We have applied time of flight secondary ion mass spectrometry (ToF-SIMS) to study lipid structural effects of stimulant drug methylphenidate on the fly brain, and to investigate the 3D chemical anatomy of C. elegans. Different distributions of various biomolecules, particularly fatty acids, eye pigment, diacylglycerides, phospholipids have been found across the fly brain. Lipid structures, particularly diacylglycerides (DAG), phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositiol (PI), are shown to dramatically alter following the administration of methylphenidate. For C. elegans, the entire worm and worm sections were imaged using 3 dimensional (3D) and 2 dimensional (2D) approaches, respectively. Significant changes in the chemical distribution were observed along the depth of the worm. In addition, correlation between 2D and 3D ion images showed different molecular structures across the worm, possible localization of the nerve ring, the cuticle, and the fluid containing space inside the worm. In addtion, we successfully used tandem MS on ToF-SIMS with a high energy 40 keV Ar4000+ gas cluster primary ion beam (GCIB) to elucidate the structures of molecular lipids in the fly brain and the worm. ToF-SIMS imaging shows great potential to elucidate chemical distributions in small invertebrate systems in relation to endogenous and exogenous effects.
- Track 1: Applications of Mass Spectrometry Track 5: Recent Advances and Development in Mass Spectrometry Track 9: Mass Spectrometry Configurations and Techniques Track 12:NMR Spectroscopy and NMR in biomedicine
Chair
Lianming Wu
GlaxoSmithKline, USA
Co-Chair
Medicharla V. Jagannadham
CSIR-Centre for Cellular and Molecular Biology, India
Session Introduction
Paul Lee
Discovery Technologies, Amgen Inc., USA
Title: High-throughput metabolic stability assays using RapidFire and Accurate-Mass Spectrometry
Biography:
Paul Lee joined Amgen Inc. in 2004. He currently leads a group of scientists and is responsible for developing biochemical and cellular assays, implement high-throughput screens and lead discovery in the early drug discovery process. Prior to joining Amgen, Paul held various positions at Pfizer, Pharmacia, and Glaxo-Wellcome for over 12 years. Paul received his Ph.D. in Pharmacology from University of Hong Kong and completed his postdoctoral training at NIH.
Abstract:
Metabolic stabilities of small molecules, in the presence of liver microsomes and/or hepatocytes, have been frequently assessed during lead optimization in the early drug discovery process. The resultant ranking is a key selection criterion for compound advancement into in vivo studies because a more metabolically stable compound will likely give rise to a longer half-life in animal studies. In vitro metabolic stability studies have been routinely performed using pooled liver microsomes from various species (human, monkey, dog, rat and mouse). The key detection technology of this assay is by monitoring the turnover of the compound in the presence of liver microsomes using liquid chromatography-tandem mass spectrometry (LC-MS). However, the bottleneck of the assay falls on the long separation time in LC. The use of RapidFire, a system consisting of a solid-phase extraction column, automated plate handler, sample injector, and software interface with MS, has significantly improved the throughput. In addition, the use of an Accurate-Mass Quadrupole Time-of-Flight (QTOF) MS system with RapidFire eliminates the need of tuning prior to sample analysis and further streamlines the MS analysis process. In this report, we describe the implement of RapidFire-QTOF system in human and rat metabolic stability assays, resulting in a throughput ten times higher than the traditional LC-MS method without compromising data quality.
Lianming Wu
GlaxoSmithKline, USA
Title: Ambient Mass Spectrometry: New Directions in Pharmaceutical Analysis
Biography:
Lianming Wu obtained a Ph.D. in mass spectrometry (MS) from Purdue University under the guidance of Prof. R. Graham Cooks and followed by Postdoc Research Fellow in MS-based proteomics at Pacific Northwest National Laboratory under the guidance of Dr. Richard D. Smith. Dr. Wu has published forty one peer-reviewed scientific papers and book chapters on the subject of mass spectrometry and gave a number of oral presentations in national/international conferences such as ACS National Meeting and PittCon. He is currently working in Global Spectroscopy, Analytical Sciences & Development, GlaxoSmithKline.
Abstract:
Ambient mass spectrometry (AMS), that allows rapid chemical analysis of untreated samples in the ambient environment, represents novel directions in pharmaceutical analysis. Because AMS is performed by direct sampling/ionization of analytes from native samples, it allows high throughput for the analysis of compounds in matrices using solvent-based desorption methods such as desorption electrospray ionization (DESI) or plasma-based methods such as direct analysis in real time (DART). As one of the most widely studied ambient ionization methods, DESI uses fast-moving solvent droplets to extract analytes from surfaces and propel the resulting secondary microdroplets towards the mass analyzer. The development of AMS takes advantage of the knowledge and experience accumulated in mass spectrometry as well as other areas of chemistry, physics and engineering. Several applications of AMS in supporting pharmaceutical product development are presented: (1) direct analysis of samples in matrix with little to no sample pretreatments; (2) MS-imaging (drug mapping in biological samples and tablet formulation as well as diagnosis of tissues); (3) conformation study of proteins and oligonucleotides; and potentially (4) investigation of salt forms of drugs in tablet or capsule formation. Manipulation of the chemistry accompanying ambient ionization can be used to further optimize chemical analysis. Extensions of the methodology to pharmaceutical analysis (such as direct analysis of biotransformation reaction mixtures in biological matrixes and trace analysis of genotoxic impurities) by reactive DESI via in-situ chemical derivatization (viz. gas-phase ion/molecule reactions) on surface are also covered.
Suneetha Achanti
Department of Pharmaceutical Analysis, Hindu College of Pharmacy, India
Title: PDAS–MSD- LC MS- Prescriptive Drug Analysis Study in Rat Plasma for Multiple Sclerosis Disease by LC-ESI/MS
Biography:
Dr. A. Suneetha, Professor & Head, Department of Pharmaceutical Analysis at Hindu College of Pharmacy, Guntur, A.P since 2001. She has completed her Ph.D at the age of 33 years from Acharya Nagarjuna University, Nagarjuna nagar. She has total 16 years of teaching and research experience. She has published more than 40 research papers in various reputed National and International journals. She served as BOS member of ANU in Pharmacy during 2010 to 2013. She is recognized as Research Supervisor from Acharya Nagarjuna University for guiding PhD scholars and she is a life member of APTI.
Abstract:
Prescribing a single drug and its administration is not sufficient in neuro diseases like multiple sclerosis. Combination therapy is growing enormously to decrease the number of medications for a single disease or their associated diseases. In clinical research estimation of concomitant drugs plays a key role to study the drug-drug interactions. The research in the current article has undertaken to provide an accurate method for evaluate the pharmacokinetic parameters of Carbamazepine (CBZ), Duloxetine (DLX), Tamsulosin (TSL) and Teriflunomide (TFM) using Doxofylline (DXF) and Ibuprofen (IBP) as internal standards (IS) in rat plasma by LC- MS when used as combination therapy. The developed bioanalytical method has been validated according to ICH guidelines. The obtained LODs and LOQs of all the drugs were adequate and may useful to perform the pharmacokinetic study in rat plasma. Based on the results, we can conclude that the present method is suitable for quantification of multiple analytes simultaneously without any interference and matrix effects. The concomitant drug analysis along with the target analyte is more advantageous than single compound analysis and also useful in drug interaction and toxicology studies. This method can also be useful in estimating the plasma samples of patients who administer these drugs.
Sermin Tetik
University of Marmara, Turkey
Title: Mass Spectrometry Technology for Biomarker Discovery
Biography:
Dr. Sermin Tetik is completed her PhD degree at the 2003 years at University of Marmara (Istanbul-Turkey). She have been working an academician at the same University, Faculty of Pharmacy in Istanbul-Turkey as associated professor since five years and project director of her research team focusing on thrombosis -hemostasis area. She has published more than 30 papers and above 50 international abstract in reputed journals and conferences, serving as an editorial board member of repute.
Abstract:
Human tissues and organs have affected depending on pathological condition of diseases. Proteins are basic functioning molecules of cells accordingly to modification intensities. Protein modification can be differences in cellular interaction, localization, activity, protein concentration, and co-/posttranslational. Protein biomarker discovery has covered some subtitles or points such as differantially expressed proteins for disease specific protein isomers and abnormal protein activity. Proteomic analysis and conclusions have allowed the potential protein biomarker discovery and thus diagnosis of disease and prognosis can be possible. MS-based proteomic has correlated between protein modification and a certain disease platform to accurately and confidently measure the proteins in sera or plasma which are low-abundance concentration. MS-based quantitative platform use for tissue samples or several body fluids of protein biomarker discovery. Similar to whole tissue samples human fluids such as gastric fluid, urine, blood have also been evaluated in the last decade studies. Intensity of modifications of proteins as post translational can also be indicative of a disease or its progression. Activity-based protein platform changes have allowed for proteome researchs of enzymatic and protein-drug interaction events.
Matthew B. O’Rourke
University of Technology Sydney, Australia
Title: Improving reproducibility and resolution in Imaging Mass Spectrometry (IMS) through the application of low cost manual sample preparation techniques
Biography:
Matthew O’Rourke currently has his PhD in submission after 2 years of candidature and is a resident at the university of Technology in Sydney Australia. He has published several papers in the field of Imaging mass sectrometry and tissue preservation and is an expert in MALDI applications. He also holds a patent in Australia for the development of proprietary MALDI based technology for the identification of microorganisms.
Abstract:
Matrix assisted laser desorption ionisation imaging mass spectrometry (MALDI IMS) is a technique that is currently seeing widespread use in both research and diagnostics for the spatial analysis of a range of biomolecules. It is currently predominantly used for the diagnostic analysis of cancer biomarkers in tissue microarrays. This requires automated sample preparation methods that need to be highly reproducible. Automated preparation methods rely on the spraying of dissolved matrix that can cause delocalisation of molecules, limiting potential image resolution to that of the smallest possible droplet size (usually >20 µm). The instruments are also expensive to purchase and require regular maintenance and servicing by qualified engineers. We therefore describe an alternate sample preparation method that is both low cost and highly reproducible, utilising sublimation and vapour recrystallisation. This eliminates the issue of delocalisation of molecules and allows for a potential resolution of ~1 µm, thereby enabling highly reproducible, ultra-high resolution images when the mass spectrometry instrumentation that can acquire at sub- µm resolution becomes available.
Medicharla V. Jagannadham
CSIR-Centre for Cellular and Molecular Biology, Hyderabad, India
Title: Chemical modification of peptides followed by mass spectrometry: Implications in proteomics
Biography:
M.V. Jagannadham is working as a senior principal scientist and project leader at the Centre for Cellular and Molecular Biology. He published 45 research papers in internationally reputed scientific journals. He trained several students, conducted meetings and workshops in proteomics. He received “Bharat Jyothi” award from India International Friendship Society, New Delhi in 2014 and “Eminent Mass Spectrometrist” award from the Indian Society for Mass spectrometry (ISMAS) in 2015. His current research Interests are proteomics, particularly in improving the de novo sequencing efficiency of peptides using MS techniques, structural and functional studied of outer membrane vesicles of Gram-negative bacteria.
Abstract:
Determination of the sequence of the peptides, obtained from the proteolysis digestion of a protein, using mass spectrometry, is a crucial requirement in proteomics. Sequence of the tryptic peptides is usually obtained by database search or by de novo sequencing. The poor spectral quality and signal to noise ratio interferes in the analysis. Different types of algorithms (MASCOT, SEQUEST and others) are used for interpretation of the mass spectral data. When the database is not available, de novo sequencing is the only way to determine the sequence. The de novo sequence can be obtained by employing different methods of fragmentation (CID, HCD, ETD and ECD), getting information on their sequence and combining this information. Even with MS instruments with high mass accuracy and speed of analysis, the reproducibility of the mass spectrometry-based proteomics is being questioned from time to time. Acetylation of peptides improved the spectral quality, exhibited by an increase in b ion intensities in the MS/MS spectra, improves the efficiency of de novo sequence and helped in validating the database search results. It is a simple reaction, which can be carried out on complex protein digests as is required in proteomics. The identification of proteins from an Antarctic bacterium Pseudomonas syringae Lz4W and other species using this strategy will be discussed.
Adrijana Leonardi
Jozef Stefan Institute/Department of molecular and biomedical sciences, Slovenia
Title: Snake venomics and antivenomics of the nose-horned viper, Vipera ammodytes ammodytes
Biography:
Adrijana Leonardi is a researcher at the Department of molecular and biomedical sciences at the Jozef Stefan Institute. She is a biochemist with a deep research interest in toxicology, protein structure and structure–function relationships. She has been studying hemostatically active components of snake venoms and other medically interesting compounds from animal venoms. Recently she started collaborations with research groups studying the interactions of nanoparticles with plasma proteins. She is mentoring graduate and undergraduate students. Her work was published in 36 papers in reputed journals.
Abstract:
Snake venoms are a deep well of natural compounds with high specificity for various biological targets, from which we draw new pharmacological leads. The nose-horned viper, Vipera a. ammodytes, is the most venomous European snake inhabiting a large part of the south-eastern Europe and Asia Minor. Hemorrhage and coagulopathies are the most pronounced effects of envenomation in humans. We used snake venomics approach, combining proteomics and transcriptomics, to survey the protein composition of the venom with the aim of discovering new pharmacologically active substances. Proteins were separated by two-dimensional polyacrylamide gel electrophoresis into 208 spots and identified by mass spectrometry using the venom gland cDNA library sequence data. In an activity-guided fractionation, hemostatically active components, effecting blood coagulation and platelet aggregation, were detected and isolated: procoagulants metalloproteases - factor IX, X and prothrombin activators, anticoagulant fibrinogenolytic serine proteinases with kallikrein-like and angiotensin I-degrading activity, C-type lectin-like proteins inhibiting vWF-dependant platelet aggregation, dimeric disintegrins inhibiting vWF-, collagen- and ADP-dependant platelet aggregation and phospholipases A2 with anticoagulant and platelet anti-aggregation activities. The only adequate and efficient treatment of snakebite envenoming is serotherapy. The term “antivenomics” describes proteomic procedure of identification of those polypeptides in the venom which possess epitopes that are weakly or not at all recognized by the standard homologous antivenom. Using antivenomics approach we aimed to deepen and broaden our efforts towards formulation of the optimal antigenic composition of the sample for immunization of animals to prepare highly effective antiserum for immunotherapy after envenomation with the nose-horned viper venom.
Athula B. Attygalle
Center for Mass Spectrometry, Stevens Institute of Technology, USA
Title: Charge-site location and structure elucidation of gaseous ions
Biography:
Attygalle is internationally recognized as a researcher and an educator of mass spectrometry and chemical ecology. He has extensive experience in conducting hands-on laboratory workshops and lectures on mass spectrometry. He has conducted mass spectrometry workshops in Malaysia, Pakistan, Brazil, Costa Rica, the Dominican Republic, Switzerland, Saudi Arabia, Germany, and India. Dr. Attygalle has published nearly 200 peer-reviewed research articles. Research reported in some of his joint publications has been singled out for coverage on numerous occasions by the media including the New York Times. Before joining Stevens, he served as the Director of the mass spectrometry facility of Cornell University from 1988 to 2001.
Abstract:
The location of the charge site is an important prerequisite to interpret fragmentation spectra of ions. Although the challenge appears seemingly simple, in reality it is not at all a trivial problem. One often assumes that the charge location can be easily predicted by knowing the gas-phase acidity or basicity of various groups present in a polyfunctional molecule. However, in reality this a very challenging problem because generalizations valid for solution-based chemistry cannot be extended directly to rationalize gas-phase phenomena. For example, there is sufficient experimental evidence, particularly from ion mobility mass spectrometry, to demonstrate that deprotonated p-hydroxybenzoic acid could exist in gas phase as mixture of carboxylate and phenoxide forms. Analogously, protonated aniline exists as mixture of nitrogen- or ring-protonated forms. Sometimes, the incipient proton that imparts the charge to a molecule is mobile. For example, methyl dihydrocinnamate initially protonates on the carbonyl group. Then the charge ambulates to the phenyl ring before finally migrating to the methoxy oxygen atom to afford a subsequent loss of methanol. Analogoulsy, The charge-imparting proton migrates to the ring before a molecule of water is eliminated from protonated benzoic caid. Using many similar examples, the methods, which include ion-molecule reactions, H/D-exchange studies, ion-mobility spectroscopy, and isotope-labeling experiments, available to elucidate intricate ion-fragmentation pathways will be presented and discussed.
- Track 6: Mass spectrometry Imaging
Session Introduction
Ângelo C. Salvador
University of Aveiro, Portugal
Title: GC×GC-TOFMS-based metabolomics as an approach to study the influence of elderberries (sambucus nigra) ripening and cultivar
Biography:
Ângelo Salvador is a PhD candidate in Chemistry, from University of Aveiro (UA), Portugal. He obtained the Master degree in Biochemistry and Food Chemistry at the same institution. He was co-author of a national patent (PT), 9 international research papers, 1 book chapter. His work was presented in international and national conferences, both oral communications (8) and in poster (11), being one awarded as the best oral communication. He was awarded with the best Master student from Food Chemistry field on UA and the best idea of the 2nd National Prize for SPME. He was part of a finalist team (8 out of 500) National Entrepreneurship Prize.
Abstract:
Great challenges on analytical chemistry and mass spectrometry were faced with the expansion of plant metabolomics due to the metabolites complexity. Volatile terpenic (C10 and C15) and norisoprenoid (mainly C13) metabolites, in particular, have similar structural backbones being the MS analysis often difficult. GC-MS is commonly used to analyse these compounds but the demands of plant complexity frequently overwhelm the capacity of 1D separation processes, leading to chromatographic co-elutions and limiting reliable MS identifications. Thus, this work presents a GC×GC-ToFMS-based plant metabolomics approach that explores the effects of ripening and cultivar on elderberries volatile terpenic and norisoprenoid metabolites. GC×GC-ToFMS was selected due to its high resolution and sensitivity being attained by employing an orthogonal separation mechanism (volatility and polarity). Further, ToFMS analyzer granted a high spectral resolution with narrow peaks (milliseconds) and spectral acquisition rates (100Hz). The structured chromatogram (compounds structurally related were positioned on similar chromatographic spaces) combined with the ion extraction chromatography increased the method specificity and sensitivity, helped to eliminate hundreds of non-target metabolites and classify unknown compounds. Sixty-six metabolites were identified being 48 reported for the first time as elderberries components. By employing chemometric tools it was observed that ripening was the variability main factor (39.8%), followed by cultivar (10.3%).
Meryem SeferinoÄŸlu
Sarayköy Nuclear Research and Training Centre, Turkey
Title: Evaluation of uncertainty components associated with alpha-particle spectrometric measurements of plutonium isotopes in bilberry
Biography:
Meryem Seferinoğlu has completed her PhD at the age of 33 years from Hacettepe University, Institute of Pure and Applied Sciences, Department of Chemistry (Turkey). She worked as a postdoc at Luleå University of Technology, Division of Physics & Division of Process Metallurgy (Sweden) and worked as a researcher at European Commission-Joint Research Center-Institute fort reference Material and Measurements. She has been working Turkish Atomic Energy Authority, Sarayköy Nuclear Research and Training center, Radionuclide Metrology Department since 2007.
Abstract:
The measurement uncertainty is an essential part of the assessment of the measurement result. Previously the uncertainty in the radioactivity measurements was defined by the standard deviation of repeated measurements or was not reported. The standard deviation in a radioactive measurement given by the square root of the number of counts reflects only the statistical uncertainty of counting. Recently, more attention is given to the evaluation of the measurement uncertainty and to the preparation of uncertainty budget. There is, however, no common procedure for the estimation of a realistic uncertainty. The ISO Guide “Guide to the Expression of Uncertainty in Measurement†sets up general rules for evaluating and expressing uncertainty in a physical measurement. Some recent European documents give further details about calculating uncertainty in calibration and in quantitative chemical analysis. Nonetheless, the application of these concepts to environmental radioactivity measurements involving radiochemistry is not as straightforward as it should be, because the evaluation of uncertainty requires the analyst to take into account all stage of the method and all possible sources of uncertainty. The main purpose of this study is to demonstrate a procedure for evaluation of the measurement uncertainty in the analysis of plutonium isotopes in bilberry using a radiochemical separation method and alpha-particle spectrometry. The steps in the analytical procedure that contribute considerably to the combined measurement uncertainty are also identified. Identification of these steps might help lower the overall uncertainty of the measurements through decreasing the uncertainties in these steps.
RuAngelie Edrada-Ebel
University of Strathclyde, United Kingdom
Title: A metabolomics approach to monitor the production of secondary metabolites in microbial symbionts and endophytes through automated dereplication of HRFTMS data
Biography:
RuAnglie Edrada-Ebel is a senior lecturer in Pharmaceutical Analysis at the Strathclyde Institute of Pharmacy and Biomedical Sciences in Glasgow since 2008. She was educated a pharmacist and received her PhD in Pharmaceutical Biology from the University of Würzburg in Germany in 1998, followed by a postdoctoral fellowship at the University of California at Santa Cruz. She heads the Natural Products Metabolomics Group at the Strathclyde Institute of Pharmacy and Biomedical Sciences and leads a series of on-going projects on the application of metabolomics to identify and biotechnologically optimize the production of bioactive secondary metabolites in marine-derived microorganisms.
Abstract:
High resolution Fourier transform mass spectrometry (HRFTMS) was employed as complimentary metabolomic tool to dereplicate chemical profiles of sponge-associated microbes. Sponges act as hosts to a diverse population of symbiotic organisms including bacteria and fungi. These symbionts offer benefits to the host including protection, nutritional benefit and support to the sponge skeleton. They have also been recognised as an important source of secondary metabolites which may have bioactivity and therefore medicinal potential. The innovative strategy involved targeted cultivation and isolation of biologically active compounds. Principal Component (PCA), Hierarchical Clustering (HCA), and Orthogonal Partial Least Square-Discriminant (OPLS-DA) analyses were used to evaluate HRFT mass spectral data of culture extracts. The results of the statistical analysis identified and validated the best culture conditions and extraction procedure which optimized the isolation of novel bioactive metabolites. Production of secondary metabolites were investigated in several of the bacterial symbionts that were isolated from marine sponges. Novel secondary metabolites were screened using high resolution mass spectrometry-based metabolomics approaches. Metabolomic profiling using HR-ESIFTMS-MS fragmentation were done at different stages of the growth phase for both solid and liquid culture media and subjected to molecular networking. Molecular networking the mass fragmentation data pinpoints functionalities that can be correlated to bioactivity and fermentation parameters as well as to predict and enhance the biosynthetic pathway involved in the production of the target secondary metabolite. Dereplication studies were accomplished by utilizing the Mzmine software with Antibase and DNP databases. The optimised method in terms of media, incubation time, and maximum production bioactive compounds are taken into account for the scale-up. Metabolomic- and bioassay-guided isolation approaches were carried out to target the compound(s) of interest. We apply a similar approach on terrestrial endophytic fungi. Endophytic fungi also represent a potential source of novel chemistry and biology.
R.K. Vatsa
Bhabha Atomic Research Centre, India
Title: Photoionization of xenon clusters: Does the increasing laser intensity help in multiple ionization ?
Biography:
Dr. R.K. Vatsadid his M.Sc. (Physical Chemistry) from Delhi University and obtained his Ph.D. under supervision of Dr. J.P. Mittal from University of Mumbai, India followed by postdoctoralstudies (Prof. J. Wolfrum) at Heidelberg University, Germany. He is currently Heading Structural ChemistrySection in Chemistry Division of BARC. He has published 140 research papers in internationally reputed journals. He is vice-president of Indian Society for Mass Spectrometry (ISMAS) and an elected fellow of National Academy of Sciences (NASI), India
Abstract:
In the photoionization of CH3I clustersby laser pulses with an intensity of 109 W/cm2 (ï¬= 532 nm), formation of energetic, multiply charged atomic ions was reported by us[1].The C+3 ions,detected in TOF-MS,require a total energy of about 100 eV for formation (starting from CH3I) and in addition they had kinetic energy of ~ 300 eV. Increasing the size of the cluster resulted in enhancement of charge state as well as kinetic energy of the ions. Similar observations were made for other molecular as well as atomic cluster systems. For a given average cluster size, a further enhancement in charge state can be achieved by increasing the intensity of ionising laser. In the case of atomic clusters, we have previously reportedformation of multiply charged xenon ions up to Xe+10 state using 8 ns, 532 nm laser pulses with anintensity of~ 5x 109 W/cm2 [2]. We now report our results on ionization of xenon clusters (of similar average size) using 30 ps pulses with an intensity of ~1012 W/cm2. The ionization wavelengths(266 and 532 nm) were selected to probe if wavelength plays any role in multiple ionization. For the case of high intensity ionization at 266 nm, a strong mass peak for Xe+, followed by a mediumintensity peak for Xe+2 and a small peak for Xe+3 were observed. For these three charged states, theabundance of given state follows Boltzmann distribution. For photoionization at 532 nm, charge state up to Xe+10 could be observed with Xe+9 being the most abundant ionic species.For the lower charge states (+2 to +7), a non- Boltzmann distribution of ionic states was observed. The main cause of such a behavior is absorption of high amount of energy from the laser pulse leading to extended depletion of lower charge state which act as feedfor higher charged states.Theseresults at 1012 W/cm2 intensity once again confirm previous observations that the total energy absorbed by the cluster is much higher at longer wavelength. During the presentation, results obtained on photoionization of Xenon clusters at 1012 W/cm2 will be compared with earlier results obtained using 109 W/cm2. The apparent absence of intensity effects on overall ionization state of xenon will be discussed taking into considerationinitial multiphoton ionization by the laser pulse and subsequent electron induced ionization.
- Track 11: Advances in Chromatography
Session Introduction
Franco Biasioli
Fondazione Edmund Mach, Italy
Title: Endogenous and exogenous volatile compounds in breath as detected by PTR-MS
Biography:
Franco Biasioli holds a degree in Physics (University of Trento, Italy) and a PhD in Physics (University of Innsbruck, Austria). He is a senior researcher at Fondazione Edmund Mach (Italy) where he set up and leads a facility for the high-sensitivity monitoring of volatile organic compound based on direct injection mass spectrometry and spectroscopic methods. His research is focused on food applications but extends also to health and environment. He has published more than 140 papers in ISI journals (h-index=26, WoS) and is serving as a reviewer both for reputed journals and agencies.
Abstract:
Among the different approaches to volatile organic compound (VOC) profiling, Proton Transfer Reaction Mass Spectrometry (PTR-MS), shows intriguing peculiarities: it is fast, non-invasive and can provide very high sensitivity. Moreover, recent technological advancements partly overcome its limitations in compound identification and quantification. Here, the use of PTR-MS for the real time measurement of the concentration of VOCs in breath is discussed with reference to two studies: i) the monitoring of exogenous flavor compounds during food consumption and ii) the monitoring of endogenous compounds as possible markers of diseases or diet. In the first study the nose-space concentration during coffee drinking has been compared with dynamic sensory methods to investigate the perceived effect of different roasting. Data indicates that nose-space concentration can be used to distinguish different coffee samples and different assessors and to identify markers of sensory quality which clearly show multimodal characteristics. In the second series of studies the breath of human and rats has been measured to assess whether VOC concentration can be related to diseases (cirrhosis and coeliac disease) or diet. Data indicates that markers of cirrhosis can be identified by breath analysis and that they are correlated with serum bilirubin while no evidence of breath markers can be found for coeliac disease if the patience follow a gluten free diet. In conclusion: PTR-MS allows the real time analysis of breath VOCs and can be used both for exogenous or endogenous compounds analysis to investigate flavor perception or markers of disease or diet.
Biography:
Dr. Nita-Lazar received her Ph.D. in biochemistry in 2003 from the University of Basel for studies performed at the Friedrich Miescher Institute for Biomedical Research, where she analyzed protein glycosylation using mass spectrometry methods. After postdoctoral training at Stony Brook University and Massachusetts Institute of Technology, where she continued to investigate post-translational protein modifications and their influence on cell signaling, she joined the Program in Systems Immunology and Infectious Disease Research, now the Laboratory of Systems Biology, in April 2009.
Abstract:
Chemotaxis and toll-like receptor (TLR) signaling in macrophages are critical to the immune response. To model these signaling networks, we are using selected reaction monitoring (SRM) to measure the absolute abundance of pathway proteins, and the resulting values are being used as pathway model parameters. RNA-seq was performed to identify expressed transcripts, and shotgun mass spectrometry was used to identify proteotypic peptides. SRM using heavy-labeled internal peptide standards was used to quantify the chemotaxis pathway proteins (1). The transcript and protein abundance values correlated strongly, and estimated protein abundance values for the entire macrophage proteome were produced. Computational modeling of the chemotaxis pathway was performed using Simmune toolset. Molecular reaction rates were constrained using a model training dataset consisting of multiple types of in vitro microscopy data. Subsequently, model testing used an orthogonal dataset from ELISA assays of RAC1-GTP and RHOA-GTP. The model produced in silico results consistent with both the training and testing datasets, and it was determined to be robust by assessing the accuracy of thousands of perturbed models (2). These findings demonstrate the feasibility and value of combining mass spectrometry-based measurements with pathway modeling for advancing biological insight. SRM assays for the canonical TLR signaling pathway and related proteins have been also been successfully developed. A preliminary model of the TLR pathway has been developed using the resulting estimated protein abundance values. The relative proteome and secretome measurements for cells stimulated with TLR ligands and the cytokine secretion values measured by flow cytometry and ELISA are being used as additional constraints for this complex model. This research was supported by the Intramural Research Program of the National Institute of Allergy and Infectious Diseases, NIH.
Namrata Singh
Indian Institute of Chemical Biology, India
Title: Fibrinolytic activity in bovine bile lipid: Application of chromatography, mass spectrometry and In-vivo wound healing assay.
Biography:
An ether extract of nine different bacterial metabolites combined with two step (ether followed by ethanol) extract of bovine bile lipid is used as an immune stimulatory drug. While characterizing the drug, we observed fibrinolytic activity in the extract through fibrinogen plate assay and fibrin zymography. Background literature emphasized major role of fibrinolytic enzymes in activating immune systems. This increased our curiosity to understand the role of these enzymes in this drug in human physiology. This fibrinolytic enzyme/s has no similarity with plasmin in terms of cross reactivity in immunoblot assay and hydrolysis of the specific substrate S-2251. In RP-HPLC analysis, the lipid extract was fractionated into several components. Interestingly, fibrinolytic activity was confined to all the fractions. To purify the enzyme, it was extracted from the lipid by aqueous buffer extraction and applied to CNBr activated fibrinogen substrate affinity column. Purified enzyme was tested for activation of complement system and wound healing through C3 binding and in-vivo wound healing assay respectively. The enzyme will be identified by mass-spectrometric analysis. Also, we propose to finger-print protein components present in bile lipid by MS analysis to have a better insight of the functionality of the lipid component of the drug.
Abstract:
Namrata Singh is an INSPIRE fellow of the DST (India). She is currently associated with CSIR-Indian Institute of Chemical Biology, Kolkata aiming for Ph.D. from the University of Calcutta and will submit her thesis soon. She is involved in characterizing an immune-stimulatory drug. She participated in the 7th HOPE meeting with Nobel Laureates as JSPS Fellow in Japan, 2015. She is a recipient of several national awards and also been felicitated for excellence in academic background. Publication of 4 papers with good citations has initiated her research career and a reviewer of applied medical research.
Somenath Ganguly
Sun Pharmaceutical Industries Ltd, India
Title: Hyphenated Mass Spectrometric techniques: Revolution in Pharmaceutical Analysis
Biography:
Somenath Ganguly has completed his PhD at the age of 28 years from Indian Institute of Technology, Kharagpur. He is the Associate director in the Analytical Research Department of Sun Pharmaceutical Industries Limited, Gurgaon. Sun Pharma is a premier generic pharmaceutical organization. He has published more than 10 papers and patents. He is having about twenty years of experiences in the pharmaceutical analytical research and responsible for solid state characterization, impurity profiling and chromatographic method development activities. He has vast experiences in regulatory and quality requirements in the pharmaceutical industries
Abstract:
In recent days hyphenated mass spectrometric techniques are becoming an essential tool in generic pharmaceutical analysis. United States and European pharmacopeia recommends methods in the general chapters and monographs. Techniques involve are LCMS/MS; GCMS; ICP MS; TGA MS etc. ICH; OSHA, EMEA , US FDA published guidelines for assessing genotoxic and elemental impurities in the active pharmaceutical ingredient( API) and finished drug products. Genotoxic impurities involves structures like Epoxides; Alkylating agents; Aldehydes; Nitro compounds; Hydrazines; Azo compounds etc. Elemental impurities arising out of the catalysts used in the API synthesis like palladium charcoal; nickel acetate, borohydrides; metal residues from reactors line like lead, arsenic, mercury and cadmium. The quantitation levels of these impurities are determined based on maximum daily intake of the drugs and in general limits are in the range of less than 50 ppm. Analytical methods need to be developed, adequately validated and transferred to QC locations as per ICH and regulatory requirements. Challenges and procedures in the analytical method development and validations process involving hyphenated mass spectrometric techniques will be discussed.
A. C. Sahayam
Bhabha Atomic Research Centre, Hyderabad
Title: Determination of cobalt compounds in functional drinks and nutritive supplements using HPLC-ICP-MS
Biography:
Dr. A. C. Sahayam completed his PhD at the age of 37 years from Osmania University, Hyderabad (India) and Postdoctoral research work at National Sun-Yat Sen University, Kaoshiung (Taiwan). He is Head, Bulk Analysis Section at National Centre for Compositional Characterisation of Materials, Bhabha Atomic Research Centre, Hyderabad. His field of expertise is analytical atomic spectrometry using ICP-OES and ICP-MS coupled with different sample introduction techniques. He has published more than 50 papers in reputed journals and serving as a reviewer for reputed journals.
Abstract:
Speciation analysis of cobalt in nutritive supplements has been carried out using HPLC and ICP-MS equipped with a membrane desolvation sample introduction system as detector. In this study, cobalt containing compounds, namely Co(II), cyanocobalamin (CN-Cbl), hydroxylcobalamin (OH-Cb1), 5’-deoxyladenosylcobalamin (Ado-Cbl) and methylcobalamin (Me-Cbl), were well separated by reversed phase HPLC with a microbore C8-HPLC column as the stationary phase. Gradient elution using ammonium acetate and acetonitrile at pH 4 allowed the chromatographic separation of all species, including strongly adsorbed Ado-Cbl and Me-Cbl, in less than 14 minutes. Detection limit was in the range of 0.007-0.031 ng Co mL-1 for various Co species. We determined the concentrations of cobalt containing compounds in selected nutritive supplements and two functional drinks. Over 91% of the total cobalt species was extracted using a 0.5% v/v HNO3 solution in a microwave field at 70oC within a period of 10 min; and the spike recovery was in the range of 94-101% for various species. The HPLC-ICP-MS results showed a satisfactory agreement with the total cobalt concentrations obtained by ICP-MS analysis of functional drinks and dissolved nutritive supplements samples.
Mohammad Azharuddin
University of Calcutta, India
Title: Functional Proteomic Approach in Dry Eye Disease
Biography:
Mohammad Azharuddin has submitted his PhD at the age of 28 years from Department of Biochemistry, University of Calcutta. He has published 6 papers in reputed journals and 3 patents filed under his name.
Abstract:
Identifying important biomarker(s) for a disease is a major challenge in clinical proteomics. In this paper we have introduced a metadata approach to classify proteomes from aqueous deficient dry eye (ADDE) disease. After initial selection of tear samples from control and ADDE subjects a disease specific proteome set was constructed using a simple set theory based reduction method. The properties for each of such protein members were then evaluated, and the properties having highest discriminatory power (between control and ADDE) were chosen. The electrostatic charge distribution and amino acid composition and to some extent overall hydrophobicity seemed to be the important discriminatory features. The ADDE patients thus showed a positive bias in their proteome as compared to control subjects. Similarly, the disease specific proteome contained lesser percentage of glycine and higher percentage of threonine. The meta-proteomic analysis thus implied that ADDE specific proteins have lesser flexibility in their conformation. The results seemed to have some correspondence with our recent finding that ADDE is a protein conformational disease having higher propensity for occurrence of aggregated proteins.
Bernhard Welz
Federal University of Santa Catarina, Brazil
Title: High-temperature high-resolution molecular absorption spectrometry – a supplementary technique to ICP-MS for the determination of halogens.
Biography:
Bernhard Welz has completed his PhD at the age of 29 years from University of Stuttgart (Germany). He worked for Perkin-Elmer for 32 years, the last 16 as Director of Applied Research. After his retirement, he went to Brazil and worked as visiting professor at several Universities. Now he is voluntary professor at the Federal University of Santa Catarina. He has published more than 280 papers in reputed journals with more than 9600 citations; he has an H-index of 54.
Abstract:
Inductively coupled plasma mass spectrometry (ICP-MS) is without doubt one of the most sensitive techniques for the determination of trace elements in a variety of matrices. However, there are also some limitations, and the most serious ones are with the determination of the halogens. Fluorine, which has an ionization energy of 17.42 eV, can essentially not be determined by ICP-MS, as the ionization energy of the plasma gas argon is only 15.76 eV. Chlorine and bromine have lower ionization energies of 12.97 eV and 11.81 eV, respectively, and can be determined by ICP-MS; however, severe matrix and memory effects can often be observed when conventional sample introduction with a nebulizer/spray chamber is used. One way out of this problem is using electrothermal vaporization (ETV) for sample introduction. High-temperature high-resolution molecular absorption spectrometry (HR-CS MAS) is ideally suited for the determination of non-metals, and can therefore be used as a supplementary technique to ICP-MS. The sample – in many cases a solid sample – is directly introduced into a small graphite tube furnace, a ‘molecule-forming reagent’ is added, and the furnace is heated to a previously optimized temperature – typically between 1500 °C and 2500 °C. At this temperature, diatomic molecules are formed in the gas phase, such as CaF, SrCl or CaBr, which exhibit a characteristic absorption spectrum, which can be used for the sensitive determination of these halogens. Plant materials, coal, copper concentrate, cosmetics and fish oil were investigated with this technique and the results compared with those of ICP-MS where possible.
Marek M Kowalczuk
University of Wolverhampton, UK
Title: New vistas in multisatge mass spectrometry for analysis of biodegradable polymers
Biography:
Marek M. Kowalczuk received his Ph.D. degree in 1984 from the Faculty of Chemistry, Silesian University of Technology, and D.Sc. degree in 1994 at the same University. He was a visiting lecturer at the University of Massachusetts in Amherst, MA, U.S.A. in 1990 and Marie Curie EU fellow at the University of Bologna, Italy. Currently, he is professor at the University of Wolverhampton, UK and at the Centre of Polymer and Carbon Materials, Polish Academy of Sciences, Zabrze, Poland. He is the author and co-author of over 130 scientific papers and a score of patents.
Abstract:
Biodegradable polymers play an important role in human life, increasingly irreplaceable one. Knowledge on the relationships between their structure, properties and function is essential for prospective applications of such materials in the areas safe for human health and environment. When the development of biodegradable polymers was in its infancy the most crucial features were concentrated on the effect of macromolecular architecture, new monomer systems, polymerization mechanisms and different polymerization techniques on final biodegradable properties. Significant efforts have been directed towards specific areas, such as mechanisms of biodegradation, biocompatibility, processing conditions and potential applications in medicine, protection of environment and agro chemistry. However, such aspects like bio-safety of biodegradable polymers or nano-safety of their composites were and still are frequently neglected. In the endeavor to safe biodegradable polymers mass spectrometry methods are of particular importance in (co)polymers analyses due to their high sensitivity, selectivity, specificity and speed. Examples of the mass spectrometry studies for sequencing of biodegradable (co)polymers with the use of multi-stage electrospray mass spectrometry (ESI-MSn) will be presented. The special emphasis will be given to the ESI-MSn applications in the synthesis of biodegradable copolyesters as well as ESI-MSn for identification of selected biodegradable polymers on the way of molecular labeling. The attempts to solve the difficult question regarding the molecular structure of biodegradable copolymers with relation to the specific area of applied research will be also discussed.
SÃlvia M. Rocha
Universidade de Aveiro, Portugal
Title: Metabolomics workflow construction for Aspergillus niger detection using advanced gas chromatography
Biography:
SÃlvia M. Rocha is assistant professor in the Chemistry Department at the Aveiro University. She is BA in Pharmaceutical Sciences (Coimbra University), and PhD in Chemistry (Aveiro University). For the last 24 years, her research was focused on the characterization of plant-derived products, prospection of bioactive compounds, and metabolomics applied to bodyfluids, plants and microbial systems. Her main skills are oriented to sample preparation and high through-put analysis based on comprehensive two-dimensional gas chromatography and mass spectrometry. She is leader of the x-Chromatography Lab (goo.gl/gHKsAi) and published over 95 SCI papers, 2 books, 7 book chapters, and 3 patent applications
Abstract:
Opportunistic infections by Aspergillus niger have increased in the last years, either in paediatric patients as adults, presenting a high mortality rate, therefore strongly suggesting the need for prevention or earlier diagnosis and treatment. Microbial metabolomics has been breaking new ground as very useful tool in several areas, including those related to microbial detection, since microorganisms produce several volatile metabolites that can be used as unique chemical fingerprints of each species, and possibly of strains. This richness of information holds the promise for diagnosing infections in situ (e.g. from body fluids, food products, environmental samples, among others), circumventing the laborious recovering of microbes or their genetic material. Microbial metabolomics studies have been mainly focused on the study of the volatile fraction by using 1D-GC. Nevertheless, the use of comprehensive two-dimensional gas chromatography (GC×GC) has revealed that sensitivity and limits of detection are improved compared to 1D-GC. Several challenges should be overcome, since microbial culturing in representative conditions, alongside the technical difficulties to identify and/or quantify trace metabolites within complex matrixes, as well as the inherent problems related to data processing are partially responsible for the paucity of information on the full volatile metabolome of common microbial pathogens. Thus, this talk aims to discuss new developments towards the establishment of a comprehensive platform for A. niger detection management, contributing to in-depth explore its exometabolome, which was studied upon different growth conditions, using a methodology based on headspace-solid phase microextraction combined with GC×GC-ToFMS, an advanced gas chromatographic based methodology with high resolution and high throughput potentialities. Partial Least Squares-Discriminant Analysis (PLS-DA) and cross validation were performed to assess both the predictive power and classification models robustness. In addition, PLS-DA-Variable Importance in Projection was applied to highlight the metabolites playing major roles in species distinction; decreasing the initial dataset to only 16 metabolites (A. niger Biomarker pattern). The data pre-processing time was substantially reduced, and an improvement of quality-of-fit value was achieved. This study goes a step further on exploring the potentialities of metabolomics for constructing A. niger omics pipeline that can be proposed as a high throughput tool towards its future detection based on a molecular biomarkers pattern.
Lee Suan Chua
Universiti Teknologi Malaysia, Malaysia
Title: Protein from edible bird’s nest samples by mass spectrometry
Biography:
Chua Lee Suan has completed her PhD in 2005 from Universiti Teknologi Malaysia. She is a registered chemist, university lecturer and team leader of research group in the analytics and validation study on natural products, particularly for metabolite profiling of herbs and honey. Till to date, she has published more than 50 international research articles and serving as editor for repute journals such as Journal of Ginseng Research and European Journal of Medicinal Plants.
Abstract:
Edible bird’s nest (EBN) is a delicacy and nutritious salivary secretion of swiftlets. It contains mostly protein and carbohydrate, and fat as a minor component. The proteome of EBN is still intensively investigated, mainly due to the recent scientific findings on its medical benefits and rejuvenation of skin complexion. In the present study, proteins from EBN samples were extracted by ultrasonic and detergent assisted techniques, as well as buffer solublization. Aqueous extraction assisted by ultrasonic energy produced the highest protein content, followed by detergent assisted method and buffer solubilization. A wide range of protein sizes ranging from 17-150 kDa, mostly water soluble proteins were detected from EBN samples based on gel electrophoresis. EBN from different locations were found to have slight variance in their electrophoretic protein profile. This could be due to the difference in food sources for swiftlets. The use of detergents such as non-ionic Triton X-100 and ionic SDS produced additional protein bands which could be membrane or transmembrane proteins. The trypsinized proteins were subjected to LC-MS/MS analysis and the mass spectra were matched to the protein database. Acidic mammalian chitinase precursor is the most abundant protein in EBN. The protein was reported to have anti-inflammatory activity against allergy and airway diseases. There are also collagen, lysyl oxidase homolog 3 and co-enzyme Qbinding proteins which are important for skin complexion. The results could explain the remarkable findings of previous investigators on the benefits of EBN consumption. These proteins are first time reported to be presence in EBN.
Marek M. Kowalczuk
University of Wolverhampton, UK
Title: New vistas in multisatge mass spectrometry for analysis of biodegradable polymers
Biography:
Marek M. Kowalczuk received his Ph.D. degree in 1984 from the Faculty of Chemistry, Silesian University of Technology, and D.Sc. degree in 1994 at the same University. He was a visiting lecturer at the University of Massachusetts in Amherst, MA, U.S.A. in 1990 and Marie Curie EU fellow at the University of Bologna, Italy. Currently, he is professor at the University of Wolverhampton, UK and at the Centre of Polymer and Carbon Materials, Polish Academy of Sciences, Zabrze, Poland. He is the author and co-author of over 130 scientific papers and a score of patents.
Abstract:
Biodegradable polymers play an important role in human life, increasingly irreplaceable one. Knowledge on the relationships between their structure, properties and function is essential for prospective applications of such materials in the areas safe for human health and environment. When the development of biodegradable polymers was in its infancy the most crucial features were concentrated on the effect of macromolecular architecture, new monomer systems, polymerization mechanisms and different polymerization techniques on final biodegradable properties. Significant efforts have been directed towards specific areas, such as mechanisms of biodegradation, biocompatibility, processing conditions and potential applications in medicine, protection of environment and agro chemistry. However, such aspects like bio-safety of biodegradable polymers or nano-safety of their composites were and still are frequently neglected. In the endeavor to safe biodegradable polymers mass spectrometry methods are of particular importance in (co)polymers analyses due to their high sensitivity, selectivity, specificity and speed. Examples of the mass spectrometry studies for sequencing of biodegradable (co)polymers with the use of multi-stage electrospray mass spectrometry (ESI-MSn) will be presented. The special emphasis will be given to the ESI-MSn applications in the synthesis of biodegradable copolyesters as well as ESI-MSn for identification of selected biodegradable polymers on the way of molecular labeling. The attempts to solve the difficult question regarding the molecular structure of biodegradable copolymers with relation to the specific area of applied research will be also discussed.
Nuran Elmacı
İzmir Institute of Technology, Gülbahçe-Urla, Turkey
Title: The Structural Analysis of b5 Type Mass Fragment Ions
Biography:
Nuran Elmacı is professor in İzmir Institute of Technology, Gülbahçe-Urla, Turkey
Abstract:
Tandem mass spectrometry (MS/MS) coupled with collision-induced dissociation (CID) is commonly used method for proteomic studies. Upon low-energy CID, protonated or multiply-protonated peptides undergo fragmentation via cleavage of amide bonds to generate N-terminal b and a and/or C-terminal y ions. Understanding the gas-phase structures of these ions and their fragmentation reactions has played a vital role for correct and reliable peptide/protein identification. It has been shown that, bn (n=2-4) ions commonly have oxazolone structure. However, for some cases b2 ions also adopt a six-membered ring; diketopiperazine structure. The formation of these two different structures entirely depends on the amino acid composition of the peptide sequence as well as peptide chain length. The macrocyclic structure of b5+ and larger b ions has been predicted for aliphatic peptides in experimental and theoretical studies. The macrocyclic structure is a problem for determining the amino acid sequence of a peptide. The memory of original primary structure is lost if reopening of the macrocycle takes place at different sites on the cycle. In such cases, the databases of computer programs which are used with the MS/MS results will be insufficient to identify peptide/protein. These databases must be improved by the detailed and comprehensive studies of the gas phase fragmentations of protonated peptides. This will lead to get reliable results for the proteomic researches. In this work, the structures of b5 ions; XA4 composed of alanine and X= X; Asn, Asp, Leu, Phe, Tyr, Cys, amino acid residues have been investigated by using molecular dynamics simulation method and quantum chemical methods. The X-residues have different type of substituent groups; aliphatic (-CH2-), aromatic (C rings), alcohol (-OH), thiol (-SH), amides (-CONH2) and carboxyl (-COOH). To examine the position influence of X residue, three types of linear peptide sequences have been used in the computational calculations for b5+ fragment; XAAAA, AAXAA, AAAAX. It has been observed that the b5 ions prefer to have a macrocyclic structure rather than linear oxazolone isomer. The macrocycle has some common features (backbone structure, proton location, H-bond etc.) for all b-ions studied in this research. The results of this work will be useful to understand the gas phase dissociation chemistry of peptides and will support and insight to the experimental studies.
Yuri E. Moskalenko
Russian Academy of Sciences. Russia
Title: Spectrum analysis applied for the study of intracranial slow volume fluctuations, recorded by coupling of noninvasive methods
Biography:
Yuri Moskalenko was completed his PhD at age 29 years from Pavlov Medical University in Leningrad. He is specialist in brain circulatory physiology, focusing interest on the experimental study of pressure-volume relations of liquid media (blood, CSF) inside cranium by biophysical approach in Sechenov Institure Academy of Sciences R.F. He found relations, responsible for structural and functional stability of brain blood supply system at different conditions, including space flight. He has published 180 papers in reputed journals and 8 books, three of them published in USA and England. Awarded by the President of R.F. title â€Honor Scientist of Russian Federationâ€.
Abstract:
Slow volume fluctuations inside cranium have known since the middle of XIX Century. Although during passed time relations of their amplitude and frequency with some important physiological indices (pCO2, pO2 in brain tissue, functional activity of some brain structures, neurosurgical stages of brain activation) have been established, that these fluctuation couldn’t use in practice due to impossibility of their objective evaluation. Recently, spectrum analysis was applied for the study of intracranial slow volume fluctuations, recorded by coupling of noninvasive methods - multifrequency rheoencephalography and transcranial dopplerography with healthy volunteers and neurological patients. Spectrum analysis of 3 min fragments of recordings, provided by “Chart 5†software with quantitation 128 kHz, have shown, that frequency ranges 0.01 – 0,3 Hz include majority of spectral peaks, reflecting intracranial slow volume fluctuations and include also a few peaks, connected with of arterial pressure fluctuations and respiration. Comparison frequencies and amplitudes of spectral peaks, reflected intracranial volume fluctuations, show, that they are characterized by hemispheric asymmetry and by difference in surface and deep brain structures. Spectrograms, taken with patients, demonstrate increased number of peaks and decreased of their amplitude. After treatment, together, with disappearing of pathogenic symptoms, spectrograms are normalized. Received data indicate, that spectrum of intracranial slow volume fluctuations reflect the systems integration, (brain blood and CSF circulation as well skull mechanics) responsible for brain circulatory-metabolic supply. It is possibly to conclude, that spectral analysis of intracranial slow volume fluctuations, demonstrate quality of system integration in mechanism, which responsible for nutrition support of brain functioning.
Jiejun Wu
Janssen R&D US, USA
Title: The Development and Applications of two Novel Metabolomic Platforms for Drug Discovery Research
Biography:
Jiejun Wu obtained his B.S. in Biology from Peking University (Beijing, China), and Ph.D. in Biochemistry from The Ohio State University (Columbus, Ohio) under the direction of Prof. Alan G. Marshall. He has worked in both academic and pharmaceutical industry and is currently a Scientific Director in Discovery Sciences of Janssen R&D US, a pharmaceutial R&D division of Johnson & Johnson. He has published more than 40 papers in reputed scientific journals including Nature, PNAS, Anal. Chem., Biochemistry, J. Biol. Chem., and J. Org. Chem., etc..
Abstract:
Metabolomics is the "systematic study of the unique chemical fingerprints that specific cellular processes leave behind", which requires the identification and quantification of the endogenous small molecules in a biological sample and data mapping to metabolic pathways. A properly measured metabolic profile provides a comprehensive picture of the underlying physiology, and the profile may be more translatable across different species – in comparison to genetic, or transcriptional, or proteomic profiles. Applications of metabolomics in pharmaceutical R&D are many and rapidly expanding, such as disease characterization, drug efficacy and toxicity monitoring, and discovering individual metabolic characteristics. Together with GC/MS, NMR, and CE/MS, LC/MS is the most widely used technique in metabolomic studies. In order to focus on the desired metabolic pathways, targeted metabolomic panels (rather than global metabolomic profiling) should be employeed, for which very specific analytical protocols are usually required to achieve maximal sensitivity and throughput. Automated data processing and data analysis software tools also play a very important role in metabolomics. In this presentation, we will highlight two novel LC/MS-based targeted metabolomic platforms developed within Janssen R&D - a high sensitivity and high throughput eicosanoids platform and a high sensitivity oxysterols panel - with their application examples to various drug discovery projects.
Sejalkumari Keshavlal Patel
Ganpat University, India
Title: Development and validation of bioanalytical method for estimation of methoxsalen in human plasma
Biography:
I have completed my PhD at the age of 28 years from Ganpat University, Gujarat, India. I am assistant professor in Quality Assurance Department, Shree S. K. Patel College of Pharmaceutical Education and Research, Ganpat University, Gujarat, India since last 10 years. I have published more than 37 papers in reputed national and international journals.
Abstract:
In this study, an attempt was made to describe and validate a liquid chromatography coupled with tandem mass spectrometry method for the quantification of methoxsalen, an antipsoriatic agent in human EDTA K3 plasma according to the current bioanalytical guidelines. The internal standard used was methoxsalen D3. The separation was performed on a Symmetry, C18, 4.6X 150 mm, 5 µm column using a mobile phase of 2 mM ammonium acetate and methanol 15:85 (v/v) with a flow rate of 0.80 ml/min. The detection of methoxsalen and the internal standard was performed in multiple reactions monitoring (MRM) mode using LC/MS/MS Mass Spectrometer with electro spray ionization, operating in positive ion mode. The human plasma samples were extracted using liquid-liquid extraction with methyl tert-butyl ether. The method shows a good linearity (R2 > 0.98), precision and accuracy over the range of 0.1-100 ng/ml methoxsalen in plasma. The recovery was between 93.85 and 105.25%. The limit of quantification was 0.1 ng/ml. The analysis required about 3.2 minutes run with retention time of drug 2.2 minutes. The proposed method was selective, sensitive, accurate and precise enough to be successfully applied to bioequivalence study.
Vasile I. Furdui
Ontario Ministry of the Environment and Climate Change, Canada
Title: Ultra-trace level speciated isotope dilution measurement of Cr(VI) using ion chromatography tandem mass spectrometry in environmental waters
Biography:
Dr. Vasile Furdui is a senior scientist with the Ontario Ministry of the Environment and Climate Change and Adjunct Professor with Ryerson University (Toronto, Canada). In 2003 Vasile received his Ph.D. degree in analytical chemistry from University of Alberta, working with Prof. Jed Harrison on “Blood Sample Preparation Using Microfluidic Platformsâ€. His recent publications are focused on trace and ultra-trace analysis of various organic and inorganic species, including perfluorinated compounds, perchlorate and hexavalent chromium using liquid or ion chromatography coupled to tandem mass spectrometry. His top five publications have received over 580 citations.
Abstract:
The reliable analysis of highly toxic hexavalent chromium, Cr(VI), at ultra-trace levels remains challenging, given its easy conversion to non-toxic trivalent chromium. The new approach demonstrates a novel analytical method to quantify Cr(VI) at low ng/L concentration levels in environmental water samples by using speciated isotope dilution (SID) analysis and double-spiking with Cr(III) and Cr(VI) enriched for different isotopes. Ion chromatography tandem mass spectrometry (IC-MS/MS) was used for the analysis of Cr(VI) as HCrO4- → CrO3-. While following a classical linear multipoint calibration curve a method detection limit (MDL) of 7 ng/L Cr(VI) was achieved, the modified SID-MS method adapted from U.S. EPA 6800 allowed for the quantification of Cr(VI) with an MDL of 2 ng/L and provided results corrected for Cr(VI) loss occurred after sample collection. The adapted SID-MS approach proved to yield more accurate and precise results than the multipoint calibration method, allowed for compensation of Cr(VI) reduction during sample transportation and storage while eliminating the need for frequent external calibration. The SID approach permitted continuous sample analysis for several days without the need for recalibration. This new developed IC-MS/MS method represents an alternative to the routinely used inductively-coupled plasma (ICP) instrumentation, IC-ICP-MS, and offers several advantages over detection with ICP-MS for Cr(VI), such as the absence of polyatomic interferences of 52Cr formed in the ICP (36Arl6O, 40Ar12C, 35Cl16OH, and 37Cl14NH) that require the use of dynamic reaction/collision cells or high-resolution double-focusing sector field instruments.
K Srinivasa Rao
Anwarul Uloom College of pharmacy, India
Title: Identification and Characterization of New Drugs Using Mass Spectrometry
Biography:
Dr K Srinivasa Rao, working as Professor & HoD at Anwarul Uloom College of pharmacy, Hyderabad, India and obtained his bachelor’s and master’s degree from Andhra University, Visakhapatnam, Andhra Pradesh (India) and Ph.D and D.Sc from Berhampur University, Berhampur, Orissa (India) in Pharmaceutical Analysis. He has completed his` Doctor of Science (D.Sc) at the age 34 years. His area of interest is in phytopharmaceutical analysis & standardization and interpretation of IR, NMR, MS Spectra’s. He has several publications of national and international repute. He is a reviewer for many journals of national and international repute. He has guided 72 M.Pharm students, and a few students are pursuing Ph.D under his guidance.
Abstract:
Mass spectrometer can be used in 3 principal ways: firstly, to measure the molecular weights with very high accuracy; from these can be deduced exact molecular formulae. Secondly, to detect within a molecule the places at which it prefers to fragment; from this can be deduced the presence of functional groups within the molecule. And thirdly, as a method for identifying drug molecules by comparison of their mass spectra with libraries of digitized mass spectra of known compounds. Molecular ion gives highly useful information about the identity of the drug molecule. Fragmentation pattern gives further information about the structure of the drug molecule. All fragment ions are, however, not of equal significance to assign the structure to a compound. Intensity of the molecular ion peak in a mass spectrum depends on the type of the compound. Mass spectra cannot be interpreted if they contain any misinformation. Unfortunately, even referred journals and carefully edited collections of the standard spectra sometimes contains spectra that fail to meet this criterion. Some compounds e.g., alcohols may fail to give a visible molecular ion peak. Judging whether or not a mass spectrum is credible is sometimes the most critical step in its interpretation. Most EI mass spectrometers in use today lack sufficient resolving power to provide accurate mass measurement for the determination of elemental composition. However, the elemental composition of an ion can sometimes be determined from the ratios of the peak intensities of the isotope peaks for that ion to the intensity of the nominal mass peak. For eg., if the M+2 peak of the parent ion looks larger than the M+1 peak, the compound might contain S, Cl or Br. When there is a larger gap and a peak at 127, iodine may be present. The intensity of the M+1 peak can be used to know the number of carbons well as nitrogen atoms. In the absence of nitrogen, the maximum number of carbaon atoms can be calculated by dividing the relative intensity of the M+1 peak by 1.1. In case nitrogen is present its contribution to the M+1 peak will amount of 0.4 X number of nitrogen atoms. This quantity must be subtracted from the measured relative intensity of the M+1 peak to know the number of carbon atoms. In this talk we will address, how to identify different drug molecules using mass spectrophomter with relevant examples.
Clemens Schwarzinger
Johannes Kepler University Linz, Austria
Title: Sample preparation in MALDI ToF Mass Spectrometry
Biography:
Clemens Schwarzinger has completed his PhD at in 2002 from the Johannes Kepler University Linz (Austria). After industrial PostDocs and being guest professor at the Petroleum Institute in Abu Dhabi (UAE) he is associate professor at the Institute of Chemical Technology of Organic Materials at the Johannes Kepler University Linz. The main research interests are polymer mass spectrometry, analytical pyrolysis and the development of new polymer systems, but also the use of mass spectrometry in gemology. He has published more than 50 papers and gave more than 120 presentations at various conferences.
Abstract:
Polymer mass spectrometry, especially MALDI-ToF MS, has gained a lot of attention in the last years because of the high amount of information that can be gained, such as molar mass distribution, repeat units, end groups, etc. But care must be taken when it comes to sample preparation, especially when the simple and fast “dried droplet†technique is used. As most people have already experienced dried droplet preparation tends to from rings of higher concentration at the outer rim, the so called “coffee ringsâ€, which results in inhomogeneous distribution of the compounds and therefore questionable results. We will show that several rather simples steps can be used to circumvent this phenomenon and how to produce reliable high quality data using dried droplets, as there are the use of ionic liquids as matrices or higher matrix concentration. The results were monitored with imaging techniques such as FTIR or mass spectrometric imaging to understand the processes necessary for a perfect sample preparation. When it comes to copolymers things are getting even worse. In this case it is mostly necessary to separate the polymer into fractions either by precipitation or by SEC. We have found that a modified Electrospray interface coupled to the SEC is a very efficient and elegant way yielding best results in terms of polymer separation and MALDI sample preparation.
Tatsuya URABE
Nishina center for Accelerator-Based Science, RIKEN, Saitama, Japan
Title: Introduction of liquid samples into high-vacuum plasma ion source for low-cost quantification of inorganic elements
Biography:
Tatsuya Urabe has completed his PhD in 2010 from Tokyo University of Marine Science and Technology and worked as a postdoctoral researcher of Japan Society for the Promotion of Science (JSPS) until 2011. Now he is working in an accelerator facility at RIKEN and developing a new mass spectrometer based on accelerator science.
Abstract:
We have been developing a mass spectrometer based on plasma ion source, ECRIS (Electron Cyclotron Resonance Ion Source). Although ECRIS is known as a large-sized ion source in accelerator facility, we succeeded to miniaturize and mount it on a portable mass spectrometer (called “miniECRIS-MSâ€) for detecting gas compounds1). The miniECRIS-MS contains a permanent magnet for an ion source, ion optical system, quadrupole mass analyzer and so on. In this poster session, we will present our ongoing project for quantifying elements in liquid samples. Comparing gas samples, liquid sample introduction into high vacuum plasma (i.e. ECRIS) is troublesome. However, it can offer great advantages over present analytical methods such as ICP-MS in terms of stability, sensitivity, freedam from polyatomic (spectral) interence, initial/running costs, and portability. Details of sample introduction system are as follows. Electrospray is used as a splaying unit of liquid samples at first stage.The solutions containing the analyte of interest are electrosprayed and gradually desolvated with the aid of heated assist gas at atmospheric pressure. Then, sample aerosol was introduced into ECRIS through differential pumping systems. Since analytes of interest are charged by electrospray (as relatively large clusters), they can be electrically extracted by ion lenses such as ion funnel in the pumping system. And they are finally reached at ECRIS and ionized (decomposed) into monvalent atomic ions for quantitative purpose. By this system some solvents (water and methanol) and inorganic elements (Na) were successfully observed, and the potential of miniECRIS-MS will be discussed in the session.
Weiwen Chen
Guangzhou University of Chinese Medicine, China
Title: UPLC-Q-TOF/MS-based screening and identification of the main alkaloids and their metabolites in rat plasma,urine and feces after oral administration of Zanthoxylum nitidum decoction
Biography:
Professor Weiwen Chen obtained his PhD in cell biology in 1988 from the Xavier-Bichat Medical School, Université Paris Diderot-Paris 7, France. He is currently the Vice Principal of Guangzhou University of Chinese Medicine. His research interests focus on curing gastric-intestinal diseases using integrated traditional Chinese and western medicine, as well as on novel drugs R & D. He has published more than 100 papers in reputed journals.
Abstract:
Zanthoxylum nitidum (Roxb.) DC (Rutaceae), called Liangmianzhen in China, is used traditionally for several medicinal purposes. Its pharmacological effect have been primarily attributed to the presence of a number of alkaloids. However, due to lack of metabolism research, its main bioactive alkaloids in vivo is still unknown. A systematic method based on ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS) technique combined with MetabolitePilot software was developed to speculate the metabolites and excretion profiles of the main alkaloids in Zanthoxylum nitidum decoction in rats plasma, urine and feces samples after oral administration of the decoction. As a result, 6 parent components and a total of 18 metabolites of 3 main alkaloids including magnoflorine (MF), α-allocryptopine (AC) and skimmianine (SA) were tentatively detected in vivo. All metabolites were detected including hydroxylated, demethylated, ketonization products and their sulfate and glucuronide conjugates. 11 metabolites were from the rat plasma, 14 from the urine and 8 from the feces. Among them, metabolites of AC and SA were reported firstly. In conclusion, the research provided useful information for further study of the pharmacology and mechanism of action of Zanthoxylum nitidum decoction in vivo.
Igor Spivak-Lavrov
Aktobe Regional State University, Kazakhstan
Title: Time-of-flight mass analyzers based on a wedge-shaped electrostatic mirror with 2d field
Biography:
Igor Spivak-Lavrov holds a degree – Doctor of Physical and Mathematical Sciences (USSR). He defended his doctoral thesis on the problems of optics of charged particles at the age of 54 years at the St. Petersburg Technical University. He is a professor in the Department of Theoretical Physics of Aktobe Regional State University (Kazakhstan). He has published more than 200 papers in reputed journals.
Abstract:
The paper deals with the wedge-shaped mirror with two-dimensional (2D) field as an orthogonal ion accelerator and a TOF mass analyzer. The inhomogeneous electric field in the mirror created without the use of meshes and blinds by changing the potentials on the boundary electrodes of mirror. Analytical expressions for the electrostatic potential of mirror field obtained. The ions are extracted from the ion source by impulse activation of the accelerating electric field. Since the ions are extracted from various points of the source, even ions with identical masses acquire different velocities during acceleration. As a result, the “rear†ions of the packet catch up with the “front†ions, and packets of ions with identical masses compress in the direction of movement. By placing the detector in a plane with the greatest compression of ion packets, one creates a time-of-flight mass spectrometer. Efficient space-time focusing allows a small mass spectrometer to achieve high resolution and sensitivity. The paper also suggests and calculates new schematics of multi-stage and multi-reflection TOF mass analyzers with wedge-shaped mirrors with 2D field.
Pankaj Pathak
All India Institute of Medical Sciences (AIIMS), India
Title: IDH1 mutations in gliomas: First series from a tertiary care centre in India with comprehensive review of literature
Biography:
His research interests span from nanoparticle detection to analytical method development involving plasma spectrometry. He published more than 50 papers, served as guest editor and reviewer for major analytical and environmental chemistry journals.
Abstract:
Mutations of the gene encoding isocitrate dehydrogenase (IDH) have been shown in a significant proportion of diffuse gliomas. These mutations are specific to gliomas and their utility for diagnosis and prognostication of these tumors is being proclaimed.IDH1 mutations in codon 132 were observed in 46% cases. The frequency was 68.8% in grade II, 85.7% in grade III and 12.8% in GBMs. R132H mutation was most frequent (84.8%). Overall frequency of these mutations was relatively higher in oligodendroglial tumours as compared to astrocytic phenotype (66.7% versus 38.4%; p = 0.06). Primary GBMs showed IDH1 mutation in only 4.4% cases. In contrast, 66.7% of secondary GBMs harboured this alteration. Patients with IDH1 mutations were significantly younger as compared to those without mutation (p = 0.001). There was a significant correlation between IDH1 mutation and TP53 mutation (p = 0.004). Although IDH1 mutation showed a positive correlation with 1p/19q deletion, the association was not statistically significant (p = 0.653). There was no correlation with EGFR amplification or PTEN deletion.
Pham Thi Tam Ngoc
University of Natural Ressource and Environnement, Vietnam
Title: Thermodynamic assessment of Cs-Te system
Biography:
Pham Thi Tam Ngoc has studied in Engineer School of Chemistry of Rennes, France in 2005. She held an engineer degree in 2010 in Pau, France. Since 2010, she has worked for the project of “Haracterization and modelling of the thermodynamic behavior of SFR fuel under iradiationâ€- the co-operation research project between Aix-Marseille University and CEA Research Center. Accomplished the PhD. During this period of time, she had prestigious opportunity to work with high-profile experts in employing CALPHAD methology. At the age of 28 she has then worked as a Lecturer at the University of Natural Ressource and Environment, Ho Chi Minh city, Vietnam, 2015.
Abstract:
The operating conditions of mixed oxide fuels (MOX) in Sodium cooled Fast Reactor (SFR) are very severe combining high temperature, high linear rating and high temperature gradient. In the nominal operating conditions of a SFR, the linear power is around 2 kW/cm3 and the temperature varies from 2500K down to 800K between the center and the periphery of the oxide fuel pellets. Due to those conditions, the volatile Fission Products (FP) like cesium and tellurium generated in the central region of the fuel pellet migrate outward through the radial cracks of the fuel matrix. At high burn up, a mixture of compounds of FP is formed in the fuel-cladding gap. This layer of FP compounds located between the external surface of the fuel pellet and the inner cladding surface is called in french the Joint Oxyde Gaine (JOG). The knowledge of phase equilibria and thermodynamic properties of the Cs–Te system is thus crucial for understanding and modeling the diffusion processes during the formation of the JOG. In this work, we present the review of phase diagram, crystallographic data and thermodynamic data of the Cs–Te binary system. The thermodynamic modeling of this system is also performed with the aid of the Thermo-Calc software. The thermodynamic descriptions derived in this work are based on the databases of Scientific Group Thermodata European (SGTE) and TBASE (ECN, Petten, Netherland) for the pure elements and the gaseous species. The compound formation and liquid mixing Gibbs energy ex- pressions are obtained by a least square optimization procedure. Comparisons between calculated and available experiments results are presented. A satisfactory agreement is achieved.
Biography:
Dr. Biljana Škrbić is full professor at the Faculty of Technology, University of Novi Sad (FoTNS) with significant experience in coordination and leadership of research projects at national and international levels; founder and head of the Centre of Excellence in Food Safety and Emerging Risks (2009-) and Laboratory for Chemical Contaminants and Sustainable Development (2015-) at FoTNS; co-editor of Central European Journal of Occupational and Environmental Medicine; vice-editor of Environmental Processes (Springer). She delivered 23 lectures as invited speaker (int. conferences) and published 107 articles in the leading international journals from SCI list with impact factors of which 9 articles are dedicated to UHPLC-MS/MS.
Abstract:
As a crude extract method followed by ultra-high performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) is a technique that has been frequently used as a routine analytical technique for mycotoxins in wheayt and corn, which applicability has been also demonstrated successfully on other matrices by the team, the intention of this presentation was to further provide the evidence on the method applicability for mycotoxin analysis of rice samples, for UHPLC–MS/MS represents the most efficient and reliable analytical technique for high efficiency isolation, unequivocal identification and accurate quantification. Thus, in present investigation, a reliable UHPLC–MS/MS method was developed and validated for simultaneous determination of twelve mycotoxins (aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, ochratoxin A, zearalenone, T-2 toxin, HT-2 toxin deoxynivalenol, citrinin, fumonisin B1 and fumonisin B2) in rice. One-step extraction using solvent mixtures of acetonitrile/water/acetic acid (79:20:1, v/v/v) without any clean-up (i.e. crude extract method) was employed for the mycotoxins extraction from rice samples. Using matrix-matched calibration it has been demonstrated that the limits of quantification were below the relevant maximum contents established by the EU regulation in rice or other cereals. The recoveries of the developed method were satisfactory between 96%–126% with relative standard deviation lower than 12%. The validated method was applied on real samples. The obtained results are used to estimate the exposure of the Serbian population to mycotoxins. To the best of our knowledge, this is the first report on occurrence of the studied mycotoxins in rice commercially available in Serbia.
MD Ahsan Habib
The University of Tokyo, Japan
Title: Development of new ion source/new desorption method for mass spectrometry
Biography:
Md. Ahsan Habib has completed his PhD in Analytical Chemistry at the age of 36 years from Saga University, Japan. Currently Dr. Habib working as a JSPS fellow at the University of Tokyo and Professor, Department of Chemistry, University of Dhaka, Bangladesh. Dr. Habib has also completed 06 years 04 post-doctoral research at Saga, Chiba, University of Yamanashi and Yokohama National University in various fields of Chemistry/Science. He has published more than 30 papers in reputed journals and presented in about 30 in national/international seminars/conferences.
Abstract:
Detection and quantification of illicit compounds at trace level is very much important for public health, security and safety. Mass spectrometry (MS) has already been demonstrated its versatility for detection and quantification of a wide range of compounds at trace level because MS can provide molecular level information of the target compounds. In MS, there are many ionization sources. So far, none of the ion source can ionize efficiently of a wide range of compounds. For example, electrospray ionization (ESI) and nano-ESI have been widely using for bio-molecules. Nano-ESI has shown better resolution than ESI but it has clogging problem. To overcome such a problem, several attempts have been taken to develop new ionization source, for example, MALDI, probe-ESI (PESI) etc. Recently we have developed new atmospheric pressure chemical ionization (APCI) using alternating current (ac) instead of dc and found as a soft ionization source for explosives detection (Habib et al., RCMS). Moreover, hollow cathode discharge (HCD) ion source has been fabricated for detection of explosives at trace level (Habib et al., RCMS). A desorption method has also been developed using an ultra-cutter to desorb highly non-volatile illicit compounds and ionized dielectric discharge (DBD) ion source, and found better limit of detection (LOD) (Habib et al., ASMS). A vacuum glow discharge ionization (vacuum-GDI) source has been fabricated for the compounds those give negative ions. A further attempt has also been taken to fabricate a hybrid ionization source for polar/non-polar, volatile/non-volatile compounds.
Biography:
Zikri Arslan received his PhD in 2000 from the University of Massachusetts at Amherst in analytical chemistry with an emphasis on applied plasma source mass spectrometry. He was a National Research Council (NRC) post-doctoral scholar between 2000 and 2002; worked at NOAA/NEFSC Howard Marine Sciences Laboratory in Sandy Hook, NJ. He also worked as research scientist (2002-2003) at the University of Maryland, Chesapeake Biological Laboratory on otolith micromilling protocols for identification of bluefin tuna stocks. His research interests span from nanoparticle detection to analytical method development involving plasma spectrometry. He published more than 50 papers, served as guest editor and reviewer for major analytical and environmental chemistry journals.
Abstract:
A chemical vapor generation (CVG) method is developed for determination of cadmium (Cd) by ICP-MS. Titanium (III) and titanium (IV) were utilized for the first time as novel additives to enhance generation of volatile Cd species, and their synergistic effects were investigated for thiourea, L-cysteine and potassium cyanide (KCN) with different mineral acids. Both Ti(III) and Ti(IV) provided the highest enhancement with KCN. The improvement with thiourea was marginal (ca. 2-fold), while L-cysteine boosted signals slightly only with Ti(III) in H2SO4. Optimum CVG conditions were 4% HCl + 0.03 M Ti(III) + 0.16 M KCN and 2% HNO3 + 0.03 M Ti(IV) + 0.16 M KCN. A 3% (m/v) NaBH4 solution was adequate for successful vapor generation. Under these conditions, sensitivity was enhanced 40-fold with Ti(III) and 35-fold with Ti(IV), that are the highest enhancement factors achieved so far in Cd vapor generation. Detection limits (3s) were 3.2 and 6.4 ng L-1 for 111Cd isotope using Ti(III) and Ti(IV), respectively. Evidence indicated that Ti(III) and Ti(IV) enhanced Cd vapor generation catalytically. Effects of transition metal ions, including Co(II), Cr(III), Cu(II), Fe(III), Mn(II), Ni(II) and Zn(II) were not significant up to 1.0 µg mL-1. Among hydride forming elements, Bi, Pb, Sb and Sn depressed signals above 0.1 µg mL-1. No interferences were observed from As(III) and Se(IV). The method was validated with determination of Cd by CVG-ICP-MS in certified reference materials, including Nearshore seawater (CASS-4), Bone ash (SRM 1400), Dogfish liver (DOLT-4), Mussel tissue (SRM 2976) and Domestic Sludge (SRM 2781).
Brigitte R. S. Pecequilo
Cidade Universitária, Brazil
Title: Self-attenuation factors in gamma-ray spectrometry
Biography:
Brigitte Pecequilo completed her PhD in 1977 from the University of São Paulo, Brazil, with a major in Nuclear Technology. She is presently Senior Research Scientist with the Environmental Radiation Laboratory at the Instituto de Pesquisas Energéticas e Nucleares, São Paulo, Brazil. Her investigations include radioecology, environmental radioprotection and radiation detection measurements. She has published more than 25 papers in reputable journals and oriented several students for Master and PhD degrees. Since 1978 she is also a partial-time professor of Advanced Topics in Nuclear Measurements at the University of São Paulo.
Abstract:
Gamma-ray spectrometry with high-resolution semiconductor detectors is currently the most widely used analytical technique for qualitative and quantitative determination of gamma-ray emmitters radionuclides. Quantification of elements relies on the correct analysys of the spectra, depending strongly on the efficiency calibration of the measurement apparatus, most often performed with aqueous standard multi-radionuclides solutions. Soever, lower energy gamma rays have less penetrating ability and tend to interact more readily with matter, so, when the efficiency calibration curve is obtained with an aqueous standard multi-radionuclides solution, a self- attenuation correction is required if samples present densities higher than the water ones. Samples such sand, soils and sediments have apparent typical densities varying from 1.2 g.cm-³ to 2.9 g.cm-³, so a correction of the efficiency curve is necessary. Self-attenuation factors are determined by Cutshall transmission technique, measuring the transmission of gamma rays through both the solid sample and an ultrapure water sample in the same geometry, using punctual sources, with gamma transitions in the range of interest. All samples are measured in the same geometry with high-resolution HPGe detectors. Self-attenuation correction factors are experimentally determined for every different sample density. A study of two hundred samples of sands and soils, allow to conclude that, to obtain more reliable analyses, one should use correction factors for the self attenuation behavior, especially in the lower part of the energy spectrum. Also, as attenuation, apart from density, strongly depends on the sample chemical composition, correction factors must be determined for each studied sample.
Mikhail Bolshov
Moscow State University, Russia
Title: Mass spectrometry with inductively coupled plasma as a technique for analysis of complex matrix.
Biography:
Dr. Mikhail Bolshov has completed PhD in 1973 from the Department of Physics, Moscow State University. 1993 has completed Dr.Sci. degree from Institute for Spectroscopy, Russian Academy of Sciences. Currently the head of the laboratory in the Institute. Since 2006 take a half time position of the Professor at the Department of Chemistry, Moscow State University. 1996 - invited Professor at the University of Florida (Gainesville), 1998-2000 the head of the laboratory in ISAS (Dortmund, Germany). Author and co-author of more than 120 publicationsand in WoS journals. Since 1988 the member of Editorial Advisory Board of Spectrochim. Acta B.
Abstract:
Mass spectrometry with inductively coupled plasma (ICP MS) takes the leading position among the instrumental techniques for elemental analysis. Main advantages of ICP-MS are: extremely high sensitivity (limits of detection down to ppt), multielement capability (detection of the elements from Li to U in one cycle from sample volume about 1 mL), high throughput, user friendly operation (not high qualification of an operator), gradually decreasing price. However as all instrumental techniques, ICP-MS suffers from spectral interferences and matrix effects – the dependence of analytical signal on the composition of a sample. These effects can strongly affect the accuracy of the analysis. Different types of interferences exhibit serious role in the analysis of the samples with complex composition e.g. biological samples (human liquids, bones, hairs), pharmaceuticals, strongly acidified resulting liquids after a solid sample digestion and so on. The strategy of accounting for or, et best, eliminating these effects depends on the types of the ICP-MS instrument. Basically two types of instruments are now widely used – quadrupole ICP MS (QMS) and Sector Field MS (SFMS). The first has low mass resolution (not better than 600), but is much cheaper. The second has resolution up to 7000, but is about 3-4 times more expensive. Evidently, the QMS instruments occupy larger part of the market. The problems of interferences are much stronger pronounced for the QMS instruments. The results obtained in my group with a simple QMS will be presented in the talk. Different strategies of the accounting and elimination of the interferences will be exemplified on the developed strategy of the analysis of bio-samples and strongly acidic liquids after rock sample digestion. In case of bio-sample analysis two techniques of a sample pretreatment were compared - 1) acid mineralization (in open or closed vessels or MW digestion), and 2) dilution by the mixture of reagents. It was found that combination of instrumental parameters optimization and use of internal standard (IS) provides adequate accuracy of the determination in blood and hair samples for crucial elements - Al, Cr, Mn, Co, Cu, Zn, As, Se, Cd, Pb. Not trivial was the finding of the analytical strategy, which allows using one and any IS independent on its mass or ionization potential. Another approach was developed for analysis of acidic liquids. The pretreatment procedure and measurement strategy will be discussed in the talk. In the end a brief overview of recent improvements of QMS instruments will be presented. These results are based on my lectures in MSU.
Grazieli Simões
Universidade Federal do Rio de Janeiro, Brazil
Title: The use of mass spectrometry and spectroscopic techniques to study sulfur containing biomolecules
Biography:
Grazieli Simões has completed her PhD at Universidade Federal do Rio de Janeiro, Brazil, in 2013 with a one-year fellowship grant at the french synchrotron light source SOLEIL. She was a researcher and a teacher at Université Paris-Sud in France from 2014 to 2016. She has recently obtained a permanent position at Universidade Federal do Rio de Janeiro in Brazil. Her last publications involve the use of different types of mass spectrometry techniques, using synchrotron radiation and electron impact techniques.
Abstract:
In recent years, there has been a large interest in the study of biological systems, namely, amino acids, peptides, and proteins, using synchrotron-based spectroscopic techniques, such as near-edge X-ray absorption fine structure (NEXAFS) or related X-ray photoelectron and X-ray emission spectroscopies. Most of the X-ray spectroscopic investigations of biologically relevant molecules, such as amino acids and their polymers, have been performed on thin organic films and liquids. The association of mass spectrometry and spectroscopic techniques has allowed for the investigation of the effects of radiation damage in sulfur containing molecules. We have performed a NEXAFS (S1s) and mass spectrometry study of solid samples of cysteine, cystine and insulin irradiated with 0.8 keV electrons. The measured mass spectra point out to processes of desulfurization, deamination, decarbonylation and decarboxylation in the irradiated biomolecules. In another study, inner-shell measurements of insulin were performed by coupling a linear ion trap mass spectrometer, equipped with an ESI source at the french synchrotron radiation facility SOLEIL. The electrosprayed insulin ions were injected, mass selected, stored in the trap, and irradiated during a well-defined period. The near-edge X-ray ion yield spectra of the 6+ charge state insulin precursor were recorded as a function of the photon energy, in the vicinity of the C1s edge.
Magnus S. Magnusson
University of Iceland, Iceland
Title: Similarity of Hierarchical Structured Clustering in Human and Neuronal interactions and on DNA: Structural and Functional analogies.
Biography:
Magnus S Magnusson, Research Professor. PhD in 1983, University of Copenhagen. Author of the T-pattern model and the corresponding detection algorithms in THEME. He has focused on real-time organization of behavior, co-directed DNA analysis, numerous papers and invited talks at numerous conferences and universities in Europe, USA and Japan. Deputy Director 1983-1988, Anthropology Laboratory, Museum of Natural History, Paris. Repeatedly invited temporary Professor at the University of Paris. Since 1991, Founder and Director of the Human Behavior Laboratory, University of Iceland. Since 1995, he is in collaboration between 24 universities based on “Magnusson’s analytical model†initiated at the Sorbonne, Paris.
Abstract:
Hierarchical structured clustering (HSC) seems characteristic of the structure of the universe balancing a small number of forces, some pulling others pushing apart, the self-similar fractal distribution of matter in the universe thus reflecting HST rather than just dispersion or clumping. HSC also characterizes a proposed pattern type, called T-pattern, detected in the temporal organization of many kinds of verbal and non-verbal human, animal and neuronal behavior and interactions and is also characteristic of the structure of DNA. Functional analogies seem to exist between the occurrence of T-patterns in “cell city†and in human cities. Structural self-similarity over many levels of biological organization suggests the possibility of a unified (mathematical, bioinformatics and system biological) approach. The T-pattern is described as a repeated hierarchical and self-similar tree structure based on a single non-terminal relation, called a critical interval (CI) relationship. The instances of a T-pattern may be seen as repeated statistical pseudo-fractal objects characterized by statistically significant translation symmetry. THEMEtm (by M.S. Magnusson, ©PatternVision Ltd) is special purpose T-pattern detection and analysis PC software using a special CI detection algorithm combined with an evolution algorithm, presented here together with results from the analysis of behavior and interactions. From the relatively slow time scale of human and animal interactions to the much faster interactions within populations of neurons in living rat brains. Analogies are discussed between T-pattern structure and functions in the „cities of proteins“ (cell city) and human cities, especially regarding specialization and the particular case of religious behavior.
Biography:
Dr. Simone König completed her PhD in Physical Chemistry at the University of Leipzig and time-out for motherhood, Simone König has done postdoctoral studies with one of the pioneers in biomedical mass spectrometry, Henry M. Fales, at the National Institutes of Health in Bethesda, MD. For over a decade, she heads the Proteomic Facility of the Interdisciplinary Center for Clinical Research at the Medical Faculty of the University of Münster, Germany. She has published more than 100 peer-reviewed papers and written and edited books. She serves as a reviewer for national and international funding organisations as well as journals and she organizes the Annual Münster Conference on Biomolecule Analysis.
Abstract:
A major problem for the acceptance of two-dimensional (2D) protein gel electrophoresis in proteome expression analysis has been the little comparability of gel images among different laboratories or even within the same working group. Now, the generation of a reference grid of marker proteins in parallel to the separation of the analyte allows the correction of protein coordinates both in x (protein isoelectric point)- and y (molecular weight)-direction. This is achieved by use of multiple fluorescent dyes for standards and samples on the same gel. A net of ~100 marker nodes spans the gel and provides reference points for triangulation. The deviation from mean is improved by an order of magnitude. The technology termed Comparative 2D Fluorescence Gel Electrophoresis (CoFGE) can be carried out in both vertical and horizontal electrophoresis instrumentation with the latter being much more comfortable. With CoFGE, archiving of coordinates of proteins identified by mass spectrometry in databases has become reality and a Repository of Gel-Separated Proteins (ReGeSeP) was generated for storing and searching protein IDs, coordinates and proteome gel images. The technology was protected (Patent application EP11167383.6, May 25, 2011), licensed, and awarded with the WWU Transfer Prize 2013/2014.
Biography:
Dr. Brigitte Simons is a market development specialist at SCIEX, specializing in metabolomics using accurate mass time-of-flight mass spectrometry solutions. Prior to working at SCIEX, Brigitte received her Ph.D. in Chemical Biology at the University of Ottawa. She then completed two research post-doctoral fellowships at the Centre for Biologics Research at Health Canada and the National Institute of Heart Lung and Blood in Bethesda MD. Brigitte also spearheads an academic partnerships program for our North American business – which listeners can browse sciex.com for more information or contact Brigitte for more information.
Abstract:
This presentation will address technologies that directly address clinical research experimentation of lipids in tissue extracts, keeping sampling and data processing throughput in mind. The human lipidome contains >100,000 different molecular species found within a small mass range; consequently, isobaric overlap makes unambiguous identification and quantitation of lipid species difficult. Herein, methods that utilize high sensitivity MS/MS techniques using a high sensitivity triple quadrupole linked LIT mass spectrometer to isolate lipid classes for identification of molecular composition and quantitation have been investigated. Furthermore, QqQ platforms can now be coupled to differential ion mobility (DMS) devices and have shown to resolve phospholipid sub-classes, triglycerides, and strikingly, the sphingomyelins (SM) were resolved from PC molecular species. The latter observation is significant considering these two classes cannot be resolved using triple quadrupole strategies alone, and their masses overlap significantly. Alterations in hepatic phospholipid composition, especially the amount of phosphatidylcholine (PC) and the ratio of PC to phosphatidylanolamine (PE) might contribute to the development non-alcoholic fatty liver disease (NAFLD). Aim was to compare PC/PE ratio in patients with NAFLD or chronic Hepatitis C (CHC) to healthy controls. This was a cross-sectional study. Liver samples were obtained from healthy controls (HC), patients with simple steatosis (SS), and steatohepatitis (NASH). Lipids were extracted from liver tissue and the total PC and PE lipid detected was quantified by tandem mass spectrometry using multiple precursor ion scanning [normalized by internal standards]. Hepatic PC/PE ratio was lower in all 3 patient groups, mainly due to a reduction in PC, which was significant in SS and NASH compared to controls. This study confirms previous animal data on PC/PE in NAFLD and represents a high throughput lipidomics platform’s utility to clinical outputs.