Day 1 :
Keynote Forum
Marek M. Kowalczuk
University of Wolverhampton, UK
Keynote: New vistas in multisatge mass spectrometry for analysis of biodegradable polymers
Time : 9:00 - 9:30 AM
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.
Keynote Forum
Eduard Rogatsky
Wadsworth Center, USA
Keynote: 25 Hydroxy Vitamin D quantitative analysis by LC/MS – assay standardization
Time : 9:30 - 10:00 AM
Biography:
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.
Keynote Forum
Regina Stabbert
Philip Morris International R&D, Switzerland
Keynote: Heterocyclic aromatic amines and their contribution to the bacterial mutagenicity of the particulate phase of cigarette smoke
Time : 10:00 - 10:30 AM
Biography:
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.
- 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.