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.
- 7.Method Development
8.Chemometric optimization
9.Applications of HPLC
10.Biochemical Applications
11.HPLC fingerprinting in Bioinformatics and Computational Biology
Session Introduction
Dusan Berek
Slovak Academy of Sciences, Slovakia
Title: Advanced liquid chromatography methods for comprehensive molecular characterization of synthetic polymers
Biography:
Dusan Berek graduated with an honour Master of Chemical Engineering from the Slovak Technical University in Bratislava. He completed his PhD Thesis at Polymer Institute, Slovak Academy of Sciences in Bratislava and Institute of Macromolecular Chemistry, Czecho Slovak Academy of Sciences in Prague. He did his DSc Thesis at Slovak Academy of Sciences and Slovak Technical University Bratislava. He is working at Polymer Institute, Slovak Academy of Sciences (PI SAS) in Bratislava since 1960. From 1992-1995 he was an elected member of the Presidium of the Slovak Academy of Sciences. He served as President of the Slovak Chemical Society, Chairman of the Czecho-Slovak and Slovak National Committee of Chemistry for the International Union of Pure and Applied Chemistry and as Member of the Board of the Federation of European Chemical Societies. He is the author of five original methods for separation of complex polymer systems; preparation and application of microporous carbon fibers. Author or co-author of two monographs on liquid chromatography and 250+ scientific papers in extenso published in refereed periodicals, and in proceedings and chapters of books, as well as 60+ patents - cited more than 2,000 times in SCI journals and monographs. He has presented over one hundred and ten invited plenary, keyand main lectures, as well as over 900 lectures short communications and poster contributions during congresses, symposia and conferences and during lecturing tours to over fourty countries. He was elected Slovak Scientist of the year 1999 and Slovak Innovator of the year 2001.
Abstract:
Most high-performace synthetic polymers exhibit besides dispersity in their molar mass also dispersity in another molecular characteristic, namely in chemical structure (composition) or in physical architecture of macromolecules. They are designated complex polymers. Typical examples of complex polymers are all kinds of copolymers, branched and functional polymers. Blends of macromolecules with distinct chemical structure or physical architecture are called complex polymer systems. Size exclusion chromatography (gel permeation chromatography), SEC/GPC is commonly used for assessment of molar mass averages and dispersities (distributions) of complex polymers and complex polymer systems. However, due to simultaneous dependence of size of macromolecules on all their molecular characteristics, as well as following limited separation selectivity, sample capacity, and low detector sensitivity, SEC/GPC can hardly give quantitative information on molar masses of complex polymers and complex polymer systems. For example, SEC/GPC even does not enable molecular characterization of binary polymer blends of components with fairly different molar masses, in which the content of the minor constituent is less than few percent. To characterize complex polymers and complex polymer systems, enthalpic retention mechanisms are to be coupled with entropy based size-exclusion retention mechanism. Of the presently utilized coupled methods of polymer LC the best known are liquid chromatography under critical conditions of enthalpic interactions, LC CC, eluent gradient liquid chromatography, EG LC and temperature gradient interaction chromatography, TGIC. The common drawbacks of the latter methods are limited to both sample recovery and capacity. LC CC permits only separation of two distinct sample constituents. Recently an alternative group of coupled LC metods was developed, namely liquid chromatography under limiting conditions of enthalpic interactions, LC LC. Similar to LC CC and EG LC, the molar mass effect is supressed also in LC LC. LC LC exhibits remarkable separation selectivity, as well as both high sample capacity and recovery. Moreover LC LC is highly robust, experimentally feasible and well repeatable. It was successfully applied to separation of chemically and physically similar macromolecules including low solubility polymers. The method enables reliable identification of very low (<1% and even <<1%) amounts of minor macromolecular admixtures in a polymer matrix. The basic principle of LC LC will be discussed in the contribution. For a comprehensive molecular characterization of complex polymer systems, two different retention mechanisms are to be applied in two separate chromatographic systems. This is the basis of two-dimensional polymer liquid chromatography, 2D-LC. A flexible approach called sequential two-dimensional polymer liquid chromatography, S2D-LC was recently developed. It consists of a combination of LC LC and SEC/GPC. The fractions produced by the LC LC column are in their entirety transferred into the SEC/GPC column for determination of average and disperity of sample. The principle of S2D-LC will be elucidated in detail and the typical examples of its application will be shown, especially separation and characterization of block copolymers that contain (small) amounts of their parent homopolymers.
Biography:
The environment is contaminated with a large number of compounds that may impact organisms in, e.g., surface waters, soil, sediments and also indoors. The complexity of the contaminant mixtures demands powerful separation and identification techniques. Whereas in the last part of the previous century, many priority pollutants were non-polar, fat soluble and bioaccumulating compounds, such as dioxins and polychlorinated biphenyls. The last decades showed emerging compounds of higher polarity such as many pesticides, perfluorinated compounds, personal care products and pharmaceuticals. These compounds not only or only marginally dissolve in lipids but are also found in surface waters and, consequently, threaten environmental and human health. In many cases, target analyses suffice to determine the concentrations of the aforementioned compounds. However, the increase in the number of chemicals being used has caused an ongoing flood of emerging compounds for which are no standards or which still need to be identified. Advanced detection systems such as modern mass spectrometry alone are not sufficient to elucidate the complex mixtures of chemicals present. The development of comprehensive two-dimensional gas chromatography (GCxGC) showed already to be extremely useful for the identification of emerging non- and medium-polar contaminants. Comprehensive two-dimensional liquid chromatography (LCxLC) has now shown to be extremely useful in the same way as GCxGC, but for more polar compounds. The significant enhancement of the peak capacity by using LCxLC, together with new, extensive libraries and software facilitates the rapid identification of toxic compounds, leading to a comprehensive chemical characterization of environmental water and sediment samples.
Abstract:
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 method development and analysis of contaminants and micro-plastics in the environment and indoor air. 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 and is among the 3000 most cited scientists in the world.
Biography:
Ferey Ludivine has completed her PhD from “Institut des Sciences et Industries du Vivant et de l’Environnement, AgroParisTech”. She is Associate Professor in the ChemBioMed U869 INSERM team, headed by Professor Philippe Barthélémy at the Faculty of Pharmacy of Bordeaux. Her research is based on the development of new analytical strategies using different separation techniques (HPLC, CE, microchip, etc.) hyphenated with various detectors (DAD, CAD, ELSD, LIF) and chemometric tools (Quality by Design, experimental designs etc.).
Abstract:
Recent pharmaceutical regulatory documents have emphasized the importance of applying Quality by Design (QbD) concept for in-depth product and process understanding to ensure product quality by the design. QbD concept has been supported by the International Conference on Harmonization guideline Q8 (R2). The goal of this present study is to demonstrate the usefulness of the QbD approach when applied to the development of separation methods in green analytical chemistry. The relevance of QbD and green analytical chemistry combination was emphasized by the case study of Active Principle Ingredient and its related substance analysis by UHPLC. Following a QbD approach, green chemistry principles were included in the analytical target profile to reduce environmental impact and minimize analyst exposure during a future routine use after the implementation of the method in pharmaceutical industry. First, a scouting phase enabled to select the stationary phase and the type of organic solvent. After applying quality risk assessment, the effects of selected critical process parameters on Critical Quality Attributes (CQAs) were evaluated through a screening design. A response surface methodology was then carried out to model CQAs as function of the retained factors and the optimal separation conditions were determined by applying desirability functions to the modeled responses. Finally, focusing at quality risk management, the design space was computed as the multidimensional subspace where the CQAs fulfilled the requirements. The method was successfully validated by the accuracy profile approach and applied to a pharmaceutical product.
Anna Poliwoda
Opole University, Poland
Title: The methods for the determination of bioactive alkaloids of hallucinogenic mushrooms
Biography:
Anna Poliwoda has completed her PhD from Opole University (Poland). Currently, she works as researcher at the Division of Analytical and Ecological Chemistry of the Opole University. In the years 2001-2002, she was a recipient of grants from Socrates-Erasmus and Swedish Natural Science Research Council in the Faculty of Analytical Chemistry, Lund University (Sweden) – research on analysis of peptides in biological samples. She has co-authored 25 scientific papers, including several chapters in monographs.
Abstract:
One of the sources of naturally occurring bioactive alkaloids with hallucinogenic properties are mushrooms of the genius such as Amanita, Psylocybe and Pholiotyna. These mushrooms produce substances (the structural analogues of tryptamine (indole) or isoxazoles derivatives) that caused the user to have a heightened state of awareness of sensory input (audio, visual, etc.). Nowadays, they are generally illegal to use in many countries, but their recreational use, especially by young people, has become an increasing problem in Europe. The twentieth-century technological achievements (such as internet) make it incredibly easy to purchase hallucinogenic mushrooms without any limitations. The main danger of fungal hallucinogens is not their toxicity, but their unpredictability actions. Therefore, their occurring and concentration level in various types of samples (e.g. mushrooms, biological fluids) must be monitored. In this presentation, the chromatographic and electrophoretic methodologies used in recent years in the analysis of fungal hallucinogenic alkaloids such as psilocybin, psilocin, ibotenic acid and muscimol in various complex sample matrices (i.e. body fluids, hallucinogenic mushrooms, etc.) will be described. The review will focused on comparison of applied analytical methods, taking into account both selectivity and efficiency of separation procedures, the used detection mode, method automation, requirements of application of sample clean-up step or derivatization. Additionally, the methods considering the analysis of fungal hallucinogens that have been developed in our laboratory will be presented too.
Gaelle Coussot
Université de Montpellier, France
Title: Proteolysis inside a coated capillary: New development for the quality control of therapeutic antibodies?
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 bioanalytical 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:
Monoclonal antibodies (mAb) represent the largest class of therapeutic molecules entering clinical studies. Due to their inherent structural complexity, the quality control (QC) of such type of molecules, necessary for their development and commercialization, represents a challenging analytical task. Common approaches in mAb QC are based on mAb reduction or proteolysis to produce a mixture of mAb fragments that are further analyzed by separation methods. These cleavage steps are usually performed off line (i.e. prior to the separation step in a distinct reactor). This is an important limitation in terms of time, reactant consumption and cross contamination by the possible formation of endogenous compounds that may further impair the quality assessment of the mAb drug. To overcome these limitations, we have first developed a fully integrated bio-analytical miniaturized methodology called D-PES (Diffusion–mediated Proteolysis and Electrophoretic Separation) for the QC of polymer-drug conjugates. With the D-PES methodology, both cleavages and separation steps are performed in-line, silica capillary being used both as nano-reactor and separation support. The methodology is based on transverse diffusion of laminar flow profile (TDLFP) mixing of reactant nano-volumes (proteolytic buffer (PB), substrate (S), enzyme (E) or reducing agent (R)) inside the capillary. Principles and results of these rapid and low operating costs microanalyses will be presented for mAb drugs. Separation optimization and mAb cleavage conditions (choice of background electrolyte, PB, ionic strength, pH…) will be discussed to demonstrate the robustness of the D-PES methodology.