Biography:

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

Abstract:

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

Biography:

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

Abstract:

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

Biography:

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

Abstract:

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

Biography:

Abstract:

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

Biography:

Abstract:

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

Biography:

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

Abstract:

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

Biography:

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

Abstract:

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

Biography:

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

Abstract:

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

Biography:

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

Abstract:

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

Biography:

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

Abstract:

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

Biography:

Abstract:

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

Biography:

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

Abstract:

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

Biography:

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

Abstract:

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

Biography:

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

Abstract:

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

Biography:

Abstract:

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

Biography:

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

Abstract:

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

Biography:

Abstract:

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

Biography:

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

Abstract:

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

Biography:

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

Abstract:

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

Biography:

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

Abstract:

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

Biography:

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

Abstract:

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