Day 1 :
- Advances in Various Chromatographic Techniques
2.Fundamentals and Advances in Liquid Chromatographic Separation Techniques
3.Advances in HPLC Instrumentation
4.Hyphenated HPLC methods
5.Chip Based Separations
6.High Efficiency and High Resolution Techniques
Session Introduction
Bogusław Buszewski
Nicolaus Copernicus University, Poland
Title: New stationary phases for separation of biologically active compounds: Synthesis, characterization and application
Biography:
Boguslaw Buszewski received PhD degree at the Faculty of Chemical Technology, Slovak Technical University in Bratislava, Czechoslovakia in 1986. In 1992, he achieved the Doctor of Sciences (habilitation) degree. In 1994, he received Nicolaus Copernicus University Professor position. In 1999, he received the title of Professor of chemistry and in 2000 received Full Professor of the Nicolaus Copernicus University in Toruń. His main scientific interests are concerned with environmental analysis, chromatography and related techniques (HPLC, SPE, GC, CZE, adsorption, and sample preparation), spectroscopy, utilization of waste and sludge and chemometrics. He is authored or co-authored 15 books, patents and more than 450 scientific papers (h= 38) and is a Member of the editorial boards of 26 national and international journals in field of analytical chemistry and separation sciences. Professor Bogusław Buszewski was President of the European Society for Separation Science (EuSSS) but now he is President of the Central European Group for Separation Sciences (CEGSS).
Abstract:
The need for selectivity determines the synthesis and development of chemically bonded stationary phases that contain various functional groups. Functional groups with specific properties, together with appropriate mobile phase qualitative and quantitative composition allow controlling separation selectivity. Silica gel and organic polymers are the most often used support for stationary phases. During preparation of the packing, organic ligands can be easily bonded to silica support which has good mechanical stability that supports efficiency of chromatographic separations. Presented study deal with the synthesis of novel group of chromatographic materials that contain different polar functional group incorporated to alkyl chains. Detailed instrumental characterization of the surface is also presented, including elemental analysis, spectroscopic techniques and chromatographic study. Solvation properties of novel materials as well as creation of electric double layer on its surface give also interesting information about surface properties. The main part of the presented work will focus of chromatographic applications of novel chromatographic materials in separation of various groups of biologically active compounds.
Jacob de Boer
VU University, The Netherlands
Title: Comprehensive multi-dimensional liquid chromatography for effect-directed analysis in effluents and surface water
Biography:
J de Boer is Director of the Institute for Environmental Studies (IVM) at the VU University in Amsterdam. He is Professor in Environmental Chemistry and Toxicology. He is Editor-in-Chief of Chemosphere. Since 2013 he is National Expert for China. His research interests are in method development and analysis of persistent organic contaminants, flame retardants and micro-plastics in the environment, indoor air and human samples. He has coordinated various European research projects and a large number of research projects for international organizations. He has published over 180 peer reviewed articles.
Abstract:
The environmentt is contaminated with a large number of compounds that may impact organisms in, e.g., surface waters. Due to complexity of the contaminant mixtures present, there is a need for powerful separation techniques with high peak capacities. The European project EDA-EMERGE aims to develop a comprehensive two-dimensional liquid chromatography (LCxLC) system linked to a 96-384 wells fraction collector, a UV detector and a high resolution time-of-flight mass spectrometer (HRToF-MS) to identify emerging toxicants in Effect-Directed Analysis (EDA). The LCxLC system was optimized using a C18 column (1.8 µm, 150x2.1 mm) as the first dimension while the selectivity of the second dimension was studied using a range of stationary phases: Phenyl-Hexyl (Coroshell, 2.7 µm, 50x4.6 mm), Pentafluorophenyl (Kinetex, 2.6 µm, 50x4.6 mm), and HILIC (1.8 µm, 50x3.5 mm). Standard compound mixtures, for example, polycyclic hydrocarbons (PAHs) and triazine, organophosphates and carbamate pesticides were used for testing. The stability of the retention times, peak widths, separation efficiency, etc. were studied. Secondly, the hyphenation of LCxLC with high resolution ToF-MS was optimized. Finally, the optimized LCxLC-UV-HRToF-MS system was used for the analysis of extracts of waste, surface, and drinking water. Extracts were prepared by either passive sampling (silicone rubber sheets) or large volume solid phase extraction (30-60 L). The C18xPFP column combination appeared to be the most suitable one. The significant enhancement of the peak capacity by using LCxLC facilitates the rapid identification of toxic compounds, leading to a comprehensive chemical characterization of environmental water samples, including toxicological evaluation of the identified compounds.
Biography:
Gerhard Kratz has completed his studies at University of Applied Sciences in Berlin. His dissertation was about planning, calculation and construction of a fluidized bed reactor. Deepening during his studies was his interest in water economy, chemical technology, plastic technology, varnish and paints, oil processing. His first positions in career were in pharmaceutical industry doing HPLC method development on various HPLC brands. He specialized in HPLC column selection and teaching customers in trouble-shooting all over the world he is supporting several internet forums for HPLC questions.
Abstract:
Liquid chromatography was defined in the early 1900s by the work of the Russian botanist, Mikhail S Tswett. His pioneering studies focused on separating compounds [leaf pigments], extracted from plants using a solvent, and in a column packed with particles. Tswett coined the name chromatography [from the Greek words chroma, meaning color, and graph, meaning writing—literally, color writing] to describe his colorful experiment. [Curiously, the Russian name Tswett means color]. Today, liquid chromatography, in its various forms, has become one of the most powerful tools in analytical chemistry. HPLC is the most used analytical testing method for pharmaceuticals and is also used in production to produce ultra-pure pharmaceuticals. For example, insulin, the last purification step is done with preparative HPLC (industrial scale chromatography), at most manufacturing processes. Stationary phases are different due to the different surface chemistry used to do the bonding. High pure Silanes are used to give each packing material its unique performance.
Sergey A Piletsky
University of Leicester, UK
Title: Robust affinity materials based on MIP nanoparticles for separation, and diagnostic applications
Biography:
Sergey A Piletsky began his research career in 1985 after graduation from Kiev State University with a MSc in Bioorganic Chemistry. In 1991 he received PhD from Kiev Institute of Bioorganic Chemistry for a work on the design of molecularly imprinted polymers (MIPs) for nucleic acids. In 1998, he joined Prof. Turner in Cranfield University and became Professor in 2002. Among his main scientific achievements were: (i) Development of computational approach for design of synthetic receptors; (ii) Development of new separation materials, assays and sensors for industrial applications and new synthetic ligands possessing biological activity; (iii) Development of solid-phase synthesis of MIP nanoparticles and automatic reactor suitable for large-scale commercial exploitation. For his scientific achievements he received a number of awards such as JSPS, DFG and Leverhulme Fellowships, the award of the President of Ukraine, Royal Society Wolfson Research Merit Award and DSc from Cranfield University in 2014. In Cranfield, he headed a major research group – Cranfield Biotechnology Centre (CBC), and in 2013 he moved his group to University of Leicester where he continues his research on MIPs for diagnostic and in vivo applications. He has published more than 280 research papers, reviews and patent applications (H index 50). In 2015 he founded a company - MIP Diagnostics which he leads as a Research Director.
Abstract:
Molecularly Imprinted Polymers (MIPs) are generic alternatives to antibodies and natural receptors in diagnostics and in separation. Here we report an efficient and flexible method for automatic synthesis of MIP nanoparticles using solid-phase automated photo/chemical reactor. Our approach requires a column-cartridge with an immobilized template docked into a thermostatic computer-controllable reactor, thereby allowing controlled manufacturing of affinity nanoparticles with narrow size distributions in the range 20-400 nm. We demonstrate synthesis of water-soluble affinity nanoparticles for various targets such as drugs, toxins, peptides, proteins and virus particles with minimal manual intervention and short reaction-cycle times. The developed reactor allows easy functionalization of nanoparticles with fluorescent, electrochemical or magnetic labels. The affinity of all synthesized nanoparticles is at the subnanomolar level which makes them suitable for practical applications in assays, sensors and in affinity chromatography. With this new development in MIP synthesis we foresee a time when the application of natural antibodies in diagnostics and affinity separation would be challenged by stable and inexpensive “plastic antibodies”.
Hiroki Kuwano
Tohoku University, Japan
Title: Micro/nano fluid handling system using surface acoustic wave
Biography:
Hiroki Kuwano received his B Eng. and M Eng. degree in Mechanical Engineering and PhD in Electrical Engineering from Tohoku University in 1975, 1977 and 1990, respectively. He was working for Nippon Telephone and Telegraph Public Corporation (NTT) for 26 years. Since 2003, he has been a Professor at Tohoku University. His research interests are nano-energy systems, sensor networks and surface acoustic wave devices. He was the recipient of the NTT President Award in 1993 and 1994 and the Best Paper Award of The Japanese Society for Precision Engineering in 1997.
Abstract:
Micro/nano fluid handling system using surface acoustic wave (SAW) is described for medical/bio, chemical synthesis; analysis and so on. We developed an atomizer and micro pump using SAW for a converged mist spray with a sustained liquid supply mechanism. The atomizer can produce a narrow mist spray of sub-millimeter width. The atomized mist spray converges spontaneously and thus requires no additional spray converging system. Key features in achieving the self-converging atomization are two-port interdigital transducers (IDTs) with a groove in between. The SAW is driven by applying an electric field on IDTs patterned on a piezoelectric substrate. The two-port IDTs drive SAWs in opposite directions toward the groove, which then separates the SAW-driven streaming on each side of the groove. Two types of self-converging atomizer were experimentally demonstrated, a line and a point atomizer. While the former uses parallel IDTs to atomize water from the groove into a thin-planar-shaped spray, the latter uses arc-shaped SAW lenses to focus the SAW into a certain point in the groove to atomize water into a narrow point spray. The developed micropumps consisted of an epoxy-based reservoir with an exhaust slot and a pair of interdigital transducers (IDTs). The liquid supply from the reservoir was controlled by SAW bursts from the IDTs. A micro tank was fabricated into the exhaust slot to stabilize the atomization. In the operational test of the micropump with water, a transfer rate of 0.24 pl per single burst was confirmed.
Karen Gaudin
Universityof Bordeaux, France
Title: Study of amphiphile molecules by RP-UHPLC-DAD/CAD for vectorization applications
Biography:
Gaudin Karen has completed her PhD from University of Paris XI (France). She is Professor and Director of a research team focusing on Analytical and Pharmaceutical developments for neglected diseases at University of Bordeaux. She has published more than 45 papers in reputed journals and serving as an Editorial Board Member of repute. The main developments concern strategies to analysis API or lipid in new pharmaceutical product using various separative techniques (RP, HILIC) and detector (CAD) eventually in green chemistry.
Abstract:
The modern pharmaceutical technology is constantly looking for new strategies of drug delivery. One of the most popular concepts is closing the therapeutic compounds in a supra-molecular nano-system formed by amphiphilic molecules. As a consequence, the chemically investigation of nano-objects requires high effective methods for qualitative and quantitative analysis of amphiphiles. Among them UHPLC can be used to characterize amphiphilic molecules and quantifying them in nano-assembly. The most common chromatographic technique first envisaged for this investigation is Reversed Phase Liquid Chromatography (RP-HPLC). It takes into consideration both parts of amphiphiles: the hydrophilic and lipophilic moieties of the molecule. The double detection using Diode Array Detector (DAD) and Charged Aerosol Detector (CAD) was selected because of the lipidic nature of this class of molecule, especially for the compounds without chromophores. This analytical development was performed with three amphiphilic molecules: DOTAU, diC16dt and DOPC which are cationic, anionic and zwitter-ionic molecules, respectively, which are synthetized for biomedical application. Several parameters were studied for their elution: solvent nature, pH, and column. The results contribute to understand behaviour of amphiphile molecules and they encourage to the further exploration this research area. Thanks to this study, purity and stability of these molecules were established in various solvents to support formulation purposes. The final perspectives of this approach will be quantification step during nano assembly achievements.
Gaelle Coussot
Université de Montpellier, France
Title: Development of analytical fingerprints for the quality control of snake venoms raw substances
Biography:
Gaëlle Coussot is an Associate Professor at the Faculty of Pharmacy of Montpellier, France. She obtained her PhD in Analytical Chemistry in 2003. She then joined for 15 months the MD Anderson Center Cancer (Houston, Texas) for a Post-doctoral position in proteomic analyses. Currently, her researches focus on the development of bio-analytical methodologies using electrophoretic, chromatographic techniques and immunoassays to characterize and/or quantify proteins and others biopharmaceuticals. Research fields include quality control of biopharmaceuticals and study of antibodies resistance to particular environmental constraints. She has published 1 patent and 14-papers in international analytical and biochemical journals.
Abstract:
Snake venoms have been of increasing interest in therapeutic research due to their rich composition in peptides and proteins. Drug formulations containing snake venom strains have already been commercialized mainly in homeopathy. Quality control (QC) of raw materials used in the composition of drugs is required for Good Manufacturing Practices. The high complexity as well as the qualitative and quantitative variations observed between animals of the same species renders snake venoms CQ difficult to implement. In this context, we have developed a fingerprint approach combining capillary electrophoresis and chemometrics for the QC of snake venom raw substances. CE is a particularly well suited technique for peptides/proteins separation. Batches of snake venom have been analyzed with many replicates. All results have been processed using several chemometrics approaches (baseline correction, signals alignment, automatic recognition of common peaks) to obtain a representative analytical trace (i.e. electropherogram) that can be used for the quality assessment of future production lots. The poster presents results obtained for Lachesis muta venom. Lachesis muta venom is widely used to treat among others thrombosis and neurological disorders.
Allan Philippe
Koblenz-Landau University, Germany
Title: Hydrodynamic chromatography coupled to ICP-MS for studying nanoparticles in complex media
Biography:
Allan Philippe has completed his PhD in 2015 at the Institute of Environmental Chemistry at Koblenz-Landau University on the interactions of natural organic matters and engineered nanoparticles and on the development of HDC-ICP-MS for environmental samples. His current focus are on the further development of analyitcal techniques for characterizing colloids in environmental matrices.
Abstract:
Investigation of colloids behaviour in the environement are highly challenging due to the lack of robust and efficient methods for sizing and quantifying engineered or natural colloids in environmental media. Hydrodynamic chromatography (HDC) coupled with ICP-MS is a practical and efficient technique for the detection of inorganic nanoparticles in complex media. The claimed advantages of the method are its versatility, its relative simplicity, high recoveries, and the possibility to combine sizing and quantitative analysis. Despite ist long history, HDC validation for environmental samples and complex particles has been investigated only recently. Especially, the effect of particle shape on the retention factor remained unaddressed, although most natural colloids are non-spherical. The recent development of single particle ICP-MS (SP-ICP-MS) made possible to use this technique in combination with HDC with a powerful synergy. This presentation reports and discusses our advances in the development of HDC-SP-ICP-MS to analyse metal containing nanoparticles (e.g. Au, Ag, TiO2, CeO2) in complex media. Using standard equipment and well characterized calibrants with different densities and coatings, we demonstrated the universality of the size calibration method for HDC-ICP-MS. Several applications of this technique to complex media demonstrate its versatility. Based on measurements of standard particles having different shapes, we propose a method to differentiate between spherical and non-spherical, including fractal agglomerates, using HDC-SP-ICP-MS. Powerful combination of detectors, versatility, and possibility to size and quantify inorganic colloids simultaneously in complex matrixes are strong arguments for a wide application of HDC in colloids analysis in environmental sciences.
Aleksandra Zatirakha
Lomonosov Moscow State University, Russia
Title: Novel high performance polymer-based anion exchangers for ion chromatography
Biography:
Aleksandra Zatirakha graduated from Chemistry Department of Lomonosov Moscow State University, Moscow, Russia, in 2008 and received her PhD in 2011. She currently holds the position of researcher at Chemistry Department, where she works in the group of design and application of new materials for HPLC separations, and also teaches Analytical Chemistry for the students. Her main interests are stationary phase design and ion chromatography, and her research is focused on the development of anion exchangers for ion chromatography and stationary phases for HILIC.
Abstract:
Since the introduction of ion chromatography (IC), this method has developed into the most powerful tool for the determination of inorganic and organic anions of low and high molecular weight. In the last decade the developments in the field of IC were aimed at increasing efficiency, sensitivity and rapidity of analysis, as well as at improving separation selectivity. Since selectivity and efficiency to the large extent depend on the surface chemistry of the stationary phase, the development of novel anion exchangers remains one of the priority tasks in modern IC. Anion exchangers for suppressed IC should be mechanically stable and withstand high pH values, which makes aromatic copolymers such as polystyrene-divinylbenzene (PS-DVB) or ethylvinylbenzene-divinylbenzene (EVB-DVB) the most suitable supports for obtaining such stationary phases. Chemical derivatization of polymer surface is considered to have certain restrictions in terms of providing high efficiency; however, some modern IC columns show that such technique can certainly be used for obtaining high performance anion exchangers. Novel chemically derivatized polymeric anion exchangers developed at Chemistry Department of Lomonosov Moscow State University are compatible with hydroxide eluents and enable efficient and selective separation of 15 anions in 30 minutes, including oxyhalides (chlorite, chlorate, bromate), weakly retained organic acids (glycolate, acetate, formate, propionate), so-called standard inorganic anions (fluoride, chloride, nitrite, bromide, nitrate, phosphate and sulphate), and carbonate. The variation of the functional ion-exchange layer’s structure provides the possibility to easily govern selectivity and efficiency of the obtained anion exchangers.
Arieh Moussaieff
Hebrew University, Israel
Title: Cell-type metabolic analysis: A new tool for understanding metabolic heterogeneity
Biography:
Arieh Moussaieff is a Senior Lecturer at the School of Pharmacy in the Hebrew University of Jerusalem. After concluding his PhD in the Hebrew University with Raphael Mechoulam, he took a Post-Doctoral position in the Weizmann Institute with Assaph Aharoni, studying cell-type metabolomics, and then a second position in INSERM, France with Daniel Aberdam, studying stem cell metabolism, a work he concluded in the Hebrew University with Yaakov Nahmias before taking a faculty position. His work has been published in the most reputed journals in the fields of metabolism, analytical chemistry and pharmacology, as well as in high impact interdisciplinary journals. His current main field of interest is the link between metabolism and cell identity.
Abstract:
Metabolic profiling provides a highly sensitive phenotype. Until recently, such analyses were performed on whole organisms or tissues or on cell cultures, providing scarce cell-specific information. This work investigates several aspects of cell-type metabolism. A metabolic map of a tissue was drawn using the Arabidopsis root as a model for cell-type metabolic analysis and a new protocol was developed, providing cell-specific metabolic profiles within a tissue. This protocol is now pursued for the cell-type analyses of developmental and cancerous processes. New insights into metabolic zonation and metabolic regulation of cell fate will be discussed.
Biography:
Lanling Chu is is currently a PhD candidate from School of Public Health of Southeast University, China. Her major is food analysis and main research field is application of nonmaterial in the analysis of food. She has published 5 papers in reputed journals.
Abstract:
Sample preparation is a crucial and tedious segment in analytical methods. The application of nano-materials in sample pretreatment is preferable and favorable to solve this bottleneck problem. In this study, several kinds of nanofibers were fabricated by electrospinning and characterized then utilized to prepare the home made extraction cartridge to extract analytes. Herein, an example was cited that a sensitive analytical method based on packed-nanofiber solid-phase extraction and high performance liquid chromatography (PFSPE-HPLC) has been developed for determination of CAP residues in milk. The optimal nanofibers, polyvinylpyrrolidone - polystyrene (PVP-PS) composite nanofibers, were selected for extraction of the CAP after comparing nanofibers and optimizing electrospinning parameters. Furthermore, condition optimization assays on pH, ionic strength, leaching and elution solvents were conducted. Under optimal conditions, the analyte was separated very well and the blank milk did not disturb the determination, and results of method validation were preferable. Additionally, PVP-PS nanofibers were suitable for extraction of quinolones and monamines in a complicated matrix. Interestingly, only several milligrams sorbent and several microlitres elution solvent were favorable to directly extract analytes in samples. This method is simple, sensitive and cost-effective, and has potential to be applied to quantitatively analyze the concentrations of polar species in samples containing complex matrix.
Rafal Gajos
Medical University of Lublin, Poland
Title: The orthogonal pressurized planar electrochromatography (OPPEC) - Its application to micro-preparative separation
Biography:
Rafal Gajos is a PhD student in Pharmaceutical Sciences. He has completed his MSc from Maria Curie-Sklodowska University, Poland, in Chemistry (Faculty of Chemistry) and Biotechnology (Faculty of Biology and Biotechnology). He is a member of the research team focusing on the new separation technique (orthogonal pressurized planar electrochromatography, OPPEC). He has published a few papers on the subject in reputed journals.
Abstract:
Orthogonal pressurized planar electrochromatography (OPPEC) is a separation technique in which mixture components are separated in an adsorbent layer of a chromatographic plate. In such separation system a mobile phase solution flows through the adsorbent layer of the chromatographic plate in the defined direction and electric field is orthogonally applied to it. Therefore, the OPPEC is a combination of both chromatographic and electrophoretic techniques into a single separation technique, in which both processes proceed simultaneously and orthogonally. This technique can be applied to analytical and micro-preparative separations, along with a collection of separated fractions. When preparative mode is applied then the solutes can be separated inthe adsorbent layer in form of bands deviatedunder different angles from the direction of the mobile phase flow. In addition the separation system of the technique can be continuously fed with a sample mixture and separated components can be continuously collected at the outlet of the system. The last feature is especially attractive for preparative separations. In the presentation we would like to focus on the preparative mode of separation withthe OPPEC technique. Based on the obtained results we want to present the possibility of using our device to separate components of test dye mixture showing different deviation angle of their bands from the direction of the mobile phase flow. In addition, we intend to present a separation mode of solutes, which show the same deviation angle of their bands from the direction of the mobile phase flow.